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Hydrogel microparticles (HMPs) are powerful tools to study and manipulate cellular behavior in 3D cell culture systems and animal models. Here, fully DNA-based HMPs are presented, whose material properties can be precisely tuned by sequence-programmable design of self-assembling DNA nanostructures. These DNA-HMPs offer control over size, stiffness, viscoelasticity and ligand presentation. They are formed by microfluidic encapsulation of two types of orthogonal DNA nanostars and a sequence-complementary DNA linker in water-in-oil droplets. By varying the valency of the DNA nanostar designs, tunable mechanical properties are achieved - spanning three orders of magnitude in Young's modulus from 30Pa$30 \,\mathrm{Pa}$ to 6.5kPa$6.5 \,\mathrm{k}\mathrm{Pa}$ with distinct viscoelastic behavior. Click-chemistry based functionalization with the small fibronectin-derived peptide cyclic-RGD (c[RGD]) enables integration into fibroblast spheroids. DNA-HMPs are stably retained within the spheroids for several days and undergo remodeling, indicating active interactions between the cells and the DNA-HMPs. Combining programmable material properties and inherent biocompatibility of DNA with straightforward functionalization and stimuli-responsiveness, these DNA-HMPs represent a versatile tool to probe and manipulate tissue behaviors in 3D cell cultures.

Providing a functional pericellular matrix (PCM) by fine-tuning the microenvironment of the articular chondrocytes (ACs) can greatly improve the outcomes of articular cartilage tissue engineering. While harvesting ACs with their PCM (chondrons) results in a low cell yield and a heterogeneous mixture of ACs and chondrons, microscale hydrogels could be used for mechanical tuning of the cell microenvironment. This may enable the use of stiffer bulk materials, improving the load-bearing capacity of the construct. This study investigates the effect of microenvironmental stiffness, independent of total construct stiffness, on the regenerative performance of ACs (ECM and PCM synthesis). Additionally, we explored articular cartilage-derived progenitor cells (ACPCs) as a possible alternative to ACs in the presented system. ACs were cultured in a soft or stiff bulk hydrogel (GelMA) or were encapsulated in soft microgels and seeded into the stiff GelMA. Constructs were seeded in an ex vivo porcine chondral defect model and cultured for 28 days with dynamic mechanical stimulation using a compression-sliding bioreactor. PCM and ECM quality were assessed through cell content analysis, immunofluorescent staining, histology, and measurements of GAG and collagen content. Cell encapsulation influenced ECM synthesis and PCM amount and completeness throughout the construct. Although the nonencapsulated groups showed stronger overall alcian blue staining, the encapsulated groups demonstrated more uniform matrix deposition throughout the depth of the tissue. Furthermore, ACPCs performed similarly to ACs. These findings suggest that the approach to differentially tune encapsulating and bulk hydrogel properties holds potential for future articular cartilage tissue engineering, and that ACPCs could be used as an alternative cell source.

<b>Background</b>: Cognitive impairment is increasingly recognized as a clinically relevant outcome in transplant medicine. We compared multidomain cognitive performance between kidney transplant (KTx) and liver transplant (LTx) recipients and explored etiology-specific differences within the LTx cohort using a standardized cognitive assessment. <b>Methods</b>: This single-center cross-sectional study included 180 adult transplant recipients, 90 KTx and 90 LTx recipients, stratified into alcohol-associated liver disease (ALD; <i>n</i> = 25) and non-ALD etiologies (<i>n</i> = 65). Cognitive performance was assessed using Addenbrooke's Cognitive Examination III (ACE-III). Median time since transplantation was 3.3 months in KTx recipients, 69.2 months in non-ALD LTx recipients, and 17.2 months in ALD LTx recipients. Group differences were analyzed using nonparametric tests with Holm correction, and independent predictors of global cognition were evaluated using multivariable linear regression adjusted for age, education, and time since transplantation. <b>Results</b>: Global cognitive performance differed modestly across groups, with mean ACE-III scores of 90.3 ± 8.7 in KTx, 89.3 ± 6.5 in non-ALD LTx, and 86.9 ± 12.5 in ALD LTx recipients (overall <i>p</i> = 0.047), although no pairwise differences remained significant after Holm correction. However, the relatively small ALD subgroup may have limited statistical power and may have led to underestimation of clinically relevant between-group differences. ACE-III scores ≤ 89 occurred in 30.0%, 47.7%, and 48.0% of participants, respectively, whereas scores < 82 were observed in 13.3%, 12.3%, and 20.0%. Verbal fluency was the lowest-performing domain across groups. Higher education and younger age independently predicted better cognitive performance; time since transplantation was not independently associated with cognitive outcome. <b>Conclusions</b>: Cognitive performance appeared broadly comparable across KTx and LTx recipients, but this finding should be interpreted cautiously given differences in post-transplant follow-up and potential residual confounding. Clinically relevant cognitive vulnerability was common across all groups, supporting structured multidomain assessment and longitudinal studies using harmonized methods.

Nutritional studies in the cattle typically focus on the rumen and its microbial environment, leaving other parts of the gastrointestinal (GI) tract largely unexplored. Thus, underlying molecular mechanisms and the responses to dietary treatment in the lower gut is poorly understood. In this study, we investigated the caecum transcriptome changes and its associated microbial communities in calves with or without artificially dosed rumen content extracted from an adult cow. Eight calves were included in the study, four received artificially dosed adult rumen content (Treated) and the rest received autoclaved rumen content as a control. We observed significant transcriptome changes in the caecum between treatments, with 1,836 differentially expressed genes (DEGs) identified. A predominant portion of the DEGs were down-regulated in the treated group, which showed significant enrichment for molecular pathways related to immune response, host response to pathogens, and inflammatory responses. For the DEGs correlated with the highest number of microbes, gene ontology analysis indicated an enrichment in pathways associated with inflammation and immune response. By comparing the microbial taxa abundance among different GI tissues collected from the same study, we observed that the same dosing strategy may lead to differential retention of the microbial community in different GI tract locations. Our work indicated that the hind gut showed robust response to artificial dosing and the caecum microbial community may interact extensively with the host to shape the development and maturity of the host immune system in early life. Furthermore, our analysis suggested that tissue-specific analysis is required to fully understand the impact of early dosing on animal performance and physiology.

Recombinant genotype I-II African swine fever virus (ASFV) strains with high virulence have been increasingly reported in China and Vietnam since 2023, raising significant concerns for disease control. In this study, we characterized the hematological, virological, pathological, and immunological dynamics in ASFV-inoculated pigs, with particular emphasis on temporal changes associated with mortality following the onset of viremia. Specific-pathogen-free pigs were intramuscularly inoculated with 1 × 10³ or 1 × 10⁵ HAD₅₀/mL of ASFV LS100 virus strain and developed acute disease characterized by high fever and severe hemorrhagic manifestations. The incubation period ranged from 3 to 5 days, with mortality occurring between 6 and 10 days post-inoculation (dpi). Viral genomic DNA was detected in blood, oral swabs, and rectal swabs as early as 2-4 dpi. Pathological examination revealed prominent necrotic skin lesions and joint swelling. Although hematological parameters and serum biochemical profiles were comparable between high- and low-dose groups, differences in viral load distribution were observed. Notably, cytokine profiling in whole blood revealed a strong and persistent upregulation of pro-inflammatory mediators, including IL-1β, IL-6, IL-12p40, TNF-α, IFN-γ, CCL2, CCL3, CCL14, CXCL9, and CXCL10, which correlated with persistent fever from 2 to 7 dpi. Collectively, these findings confirm that naturally occurring recombinant genotype I-II ASFV strains are highly virulent and capable of inducing severe systemic inflammation. Their continued circulation poses substantial challenges for ASF control and prevention in Vietnam and threatens the global swine industry.

Testicular torsion is a urological emergency that requires prompt recognition and intervention. Although it most commonly affects two age groups, including neonates and adolescents, it can occur at any age. Risk factors to consider include anatomic abnormalities, trauma, testicular tumors, and family history. Typical clinical presentation involves an acute onset of severe scrotal pain, a high-riding testicle, and an absent cremasteric reflex. We report an unusual case of a 63-year-old man who presented with a two-week history of progressive scrotal pain and swelling and was found to have concurrent testicular torsion and a large scrotal abscess. This atypical and delayed presentation highlights the diagnostic challenges of testicular torsion in older adults.

No abstract available.

<h4>ABSTRACT</h4> The leptomeninges (arachnoid and pia) are the inner layers of the meninges that envelope the brain. They define the borders of the subarachnoid space (SAS) where cerebrospinal fluid (CSF) circulates. Leptomeninges act as the barrier tissue and contribute to fluid exchange and immune function of the brain. These processes are identified as key factors in the development of neurodegenerative diseases. The extracellular matrix (ECM) is a critical structural component of leptomeninges. However, it remains insufficiently characterized in health and disease. Here, we performed complete proteomic profiling of ECM proteins (matrisome) of human leptomeninges from individuals with Alzheimer’s disease (AD) and cognitively normal controls (CN) using sequential biochemical fractionation coupled with mass spectrometry. We resolved leptomeningeal matrisome based on the biochemical properties reflected by the protein solubility. This approach identified 211 ECM proteins in human leptomeninges and revealed disease-associated shifts in leptomeningeal solubility consistent with altered matrix organization and signaling. We found that ECM-linked regulators and secreted factors extracellular sulfatase SULF2, antithrombin-III (SERPINC1), secreted frizzled-related protein 3 (sFRP3) and integrin beta-5 (ITGB5), which is a receptor for fibronectin, were differentially expressed in AD leptomeninges based on the disease status. Seven ECM proteins were identified only in AD samples, and two ECM proteins in CN samples. Pathway enrichment implicated TGF-β and Wnt-related signaling and coagulation as critical for leptomeningeal function. Our findings provide the first comprehensive proteomic characterization of the human leptomeningeal matrisome and establish a biochemical framework for investigating how meningeal matrix remodeling contributes to AD.

Chronic stress induces structural and functional changes in the brain, increasing susceptibility to major depressive disorder and other mental illnesses. Myelination deficits are a key pathological feature of stress-related disorders, yet the molecular mechanisms linking chronic stress to oligodendrocyte dysfunction remain poorly understood. Here, we used single-nucleus multiome sequencing to map gene expression and chromatin-accessibility remodeling in the hippocampus of mice exposed to chronic unpredictable mild stress (CUMS), a model that simulates key features of human daily stressors. CUMS induced broad molecular reprogramming across hippocampal cell populations, with stress-responsive gene networks significantly enriched for depression-associated genes. The oligodendrocyte lineage showed heightened vulnerability, with CUMS preferentially disrupting immature OPC/intermediate states and impairing OPC migration, OPC-to-ODC lineage progression, intercellular communication, and myelination. Integrated multiomic analysis identified stage-specific cis-regulatory elements and stress-sensitive gene regulatory networks, converging on SOXD transcription factors, particularly SOX5 and SOX6, as key regulators of OPC dysfunction. SOX6 ChIP-seq confirmed direct SOX6 binding at regulatory elements associated with genes controlling OPC morphogenesis, migration, and glutamatergic signaling. CUMS reduced SOX6 protein levels and SOX6-associated regulatory network activity in OPCs, whereas OPC-specific enhancer-driven restoration of SOX6 rescued stress-induced defects in OPC migration and myelination. Together, these findings define a stress-sensitive SOXD/SOX6 regulatory mechanism linking chronic stress to oligodendrocyte lineage dysfunction and myelination deficits, identifying SOX6 as a functional regulatory node with therapeutic potential for stress-related brain pathology.

<h4>Purpose</h4>Acute Myeloid Leukemia (AML) is driven by complex interactions between genetic mutations and epigenetic dysregulation. While alterations in chromatin modifiers are frequent, the precise downstream transcriptional networks they enable and how these networks execute the leukemogenic program remain incompletely defined.<h4>Methods</h4>We employed an integrative bioinformatics strategy. Transcriptomic data from GSE84881 (AML stromal cells) and GSE9476 (AML blasts) identified differentially expressed genes, refined via GeneCards and CellMarker to a 32-gene AML signature. Functional enrichment (GO/KEGG) and protein-protein interaction (PPI) network analyses followed. Core hubs were validated for spatial (single-cell t-SNE) and subtype-specific expression using the Hematologic Malignancy database. Perturbation analysis (GPSAdb2.0 BioTrigger) expanded the network, with pathway enrichment on responsive genes.<h4>Results</h4>The 32-gene signature enriched strongly in hematopoietic differentiation and unexpectedly in cross-lineage developmental pathways (e.g., gland, epithelial development). PPI topology revealed nine hubs: AFF1, TAL1, IKZF1, GATA1, NOTCH1, BCL2, IL1B, IRF4, ZAP70. Single-cell t-SNE showed distinct, non-overlapping localization patterns among AML subpopulations; box plots demonstrated marked expression heterogeneity across 26 molecular subtypes. Perturbation of these hubs generated a 500-gene set whose KEGG enrichment highlighted three interconnected layers: (i) Polycomb repression and ATP-dependent chromatin remodeling (epigenetic gatekeepers), (ii) FoxO signaling, cell cycle, and senescence (core oncogenic pathways), and (iii) broad cancer hallmarks including endocrine resistance and diverse solid tumor pathways.<h4>Conclusion</h4>We propose a hierarchical pathomechanism: synergistic dysfunction in chromatin remodeling and Polycomb-mediated repression establishes a permissive epigenomic landscape, enabling activation of an oncogenic transcriptional network (centered on AFF1, TAL1, IKZF1, GATA1). This network then hijacks TP53/FoxO signaling to drive cell cycle escape, apoptosis resistance, and metabolic adaptation. Our findings unify disparate molecular lesions into a coherent axis and suggest new therapeutic nodes.

The NRF2 pathway has emerged as a central regulator of cellular redox homeostasis, coordinating the expression of a broad array of genes that protect cells from oxidative and electrophilic stress. In the context of cancer, NRF2 has been recognized as a key driver of chemoresistance, as its sustained activation enhances antioxidant defenses, detoxification pathways, and metabolic adaptation, thereby promoting tumor cell survival under therapeutic stress. Beyond its canonical role in redox regulation, NRF2 also orchestrates the expression of multiple genes involved in ferroptosis, a non-apoptotic, iron-dependent form of cell death that has recently gained attention as a promising strategy to overcome drug resistance. Mechanistically, NRF2 modulates ferroptosis through several interconnected pathways, including the regulation of glutathione biosynthesis, lipid metabolism, and iron homeostasis, yet its impact is highly context-dependent and can vary according to cell type and metabolic state. In this review, we provide an overview of the interplay between NRF2 and ferroptosis, tracing the historical development of this network and highlighting the pivotal roles of specific NRF2 targets in controlling ferroptotic susceptibility. Finally, we discuss how targeted modulation of NRF2 may influence ferroptosis, offering a potential avenue for the design of innovative therapies aimed at selectively eradicating resistant tumors.

Genome-wide association studies (GWAS) have identified over 200 genetic risk loci for breast cancer, yet their target genes remain largely unknown. We conduct multi-ancestry transcriptome-wide association studies (TWAS) to discover potential breast cancer susceptibility genes. We develop ancestry-specific genetic models to predict levels of gene expression, alternative splicing, and 3' UTR alternative polyadenylation using genomic and transcriptomic data from 652 normal female tissue samples and apply these models to GWAS data of 178,534 cases and 248,300 controls for association analyses. We identify 290 genes associated with breast cancer risk, including 103 previously unreported and 46 not located at known GWAS loci, and 39 genes show distinct associations with breast cancer risk by estrogen-receptor status. Single-cell RNA sequencing and in vitro experiment data provide additional functional evidence for 169 genes. These genes are enriched in pathways implicated in breast carcinogenesis. Our study uncovers insights into breast cancer genetics and biology.

Heteroresistance (HR) is an antibiotic resistance phenotype characterized by the presence of rare resistant subpopulations (frequency ≈ 10^-7 to 10^-4) within a main susceptible bacterial population. During antibiotic exposure, these subpopulations can be enriched and cause treatment failure. Standard antibiotic susceptibility testing (AST) often fails to detect HR, and the current gold-standard population analysis profile (PAP) test is labor-intensive and time-consuming. We present a digital phenotyping approach combining droplet microfluidics with image texture to detect HR from clinical isolates, including Gram-negative (Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter baumannii) and Gram-positive (Staphylococcus aureus) bacteria isolated from bloodstream infections. Our method achieves detection at subpopulation frequencies as low as 10^-6 in 12 to 30 h, depending on bacterial species, which is faster than the PAP test, together with single-cell resolution and high-throughput. This computationally assisted microfluidic platform enables rapid and accurate identification of HR, representing a step toward targeted antibiotic therapy in critical infections.

<h4>Background</h4>Adaptation is one of the key processes of animal domestication, environmental pressures will leave footprints in the genome. Geese are widely distributed across multiple geographical conditions with distinct adaptations. However, few reports have focused on the environmental adaptability of geese. Moreover, the key environmental drivers that trigger local adaptation and its genetic mechanisms are still unknown. To this end, 35 agro-climatic variables of 257 geese from 14 Chinese breeds were obtained, the key environmental drivers and its genetic mechanism were elucidated by combining the genome data.<h4>Results</h4>Five key environmental drivers, elevation, water scarcity (BIO14), two variables as proxies of food availability (Grass and Landuse), and one variable as proxy of disease resistance (Forest) were identified after dissecting 35 agro-climatic variables by using ecological niche modeling (ENM). Integrating five key environmental drivers with single-nucleotide polymorphism variations, we conduct genome-wide selection signature analyses and genome-environment analysis. This analysis identified 1984 key genes likely integral to the environmental adaptation and mainly enriched in MAPK signaling pathways and metabolic pathways. Allele frequency distribution and selective sweep analysis revealed that some genes play crucial roles in environmental adaptation, including SATB1, NTRK2 and CTIF involved in adapting to the high-altitude hypoxic environments; ARMH3 and BDH2 participated in adapting to water deficiency; COL5L1, ULK4 and PI4KA associated with disease resistance, and other genes involved in metabolic adaptations including IQSEC1, TBC1D14, GRK5, FGGY and LRP1B, etc. Notably, we noticed that feather colour differentiation in geese might be driven by human agricultural practices, with LRP1B involved in this process.<h4>Conclusions</h4>Overall, our study elucidated the five key environmental drivers and their genetic mechanisms. The candidate genes related to environmental adaptability offer new insights into genetic resource development and breeding strategies of Chinese indigenous geese.

<h4>Background</h4>Bipolar disorder (BD) is a highly heritable condition characterized by recurrent shifts between manic and depressive states. Here we investigated the potential involvement of the habenula because it plays a central role in negative affect and behavioral regulation.<h4>Methods</h4>We investigated bilateral habenular volume and seed-based resting-state functional connectivity in a discovery cohort (78 BD, 102 controls) and an independent replication cohort (72 BD, 85 controls). Associations among habenular features, clinical symptoms, and molecular correlates were examined by integrating pathway-specific polygenic risk scores and brain-wide gene expression data from the Allen Human Brain Atlas.<h4>Results</h4>Across both cohorts, BD was associated with reduced bilateral habenular volume and increased rs-FC between the habenula and right precentral gyrus. Habenular volume correlated positively with severity of mania symptoms and negatively with severity of symptoms of anxiety and somatization. Polygenic risk scores linked the altered volume to dopaminergic pathways and altered connectivity to serotonergic pathways, while transcriptomic data linked the altered connectivity to changes in expression of synaptic membrane structures, transporter complexes, and other proteins involved in synaptic transmission.<h4>Conclusions</h4>Structural, functional and transcriptomic data identify the habenula as a critical neural hub in BD and therefore important for understanding pathogenesis and clinical manifestations.<h4>Clinical trial number</h4>Not applicable.

This all-case post-marketing surveillance study in Japan evaluated the safety and effectiveness of romiplostim, a thrombopoietin receptor agonist, in patients with aplastic anemia (AA), a rare disease characterized by pancytopenia. All patients with refractory AA who initiated romiplostim from June 2019 to September 2021 were registered. Romiplostim was administered subcutaneously once weekly for 52 weeks. Observation lasted 52 weeks or until 4 weeks after discontinuation. Occurrences of bone marrow fibrosis and hematopoietic tumors were monitored for up to 2 years after romiplostim initiation. Data from 470 patients at 220 institutions were analyzed. The mean age ± standard deviation was 66.27 ± 16.89 years, and 43.40% were male. Romiplostim was discontinued in 219/470 patients (46.60%). Adverse events (AEs) occurred in 57.02% of patients, and adverse drug reactions (ADRs) occurred in 18.09%. AEs of special interest classified as ADRs included thromboembolism (1.49%), hematopoietic tumor (1.28%), and hemorrhage (0.46%). During follow-up, hematopoietic tumors as ADRs were observed in 2/310 patients (0.65%). Hematological response was achieved in 257/327 patients (78.59%) at 27 weeks and 205/244 patients (84.02%) at 52 weeks. Platelet transfusion independence or reduced requirement was achieved in 164/213 patients (77.00%) at 27 weeks and 133/160 patients (83.13%) at 52 weeks, while red blood cell transfusion independence or reduced requirement was achieved in 194/247 patients (78.54%) and 162/186 patients (87.10%), respectively. Long-term romiplostim treatment was safe and effective in patients with refractory AA in a real-world setting. Registration: University Hospital Medical Information Network (UMIN000056465), 16 December, 2024.

Sepsis accounts for nearly 20% of global mortality, with antibiotic resistance worsening clinical outcomes. Rapid antibiotic administration and accurate pathogen identification remain crucial. It is well now known that extracellular vesicles (EVs) from human cells and bacterial membrane vesicles (bMVs) play a central role in the interaction between host and pathogen and represent promising biomarkers for early infections. This study investigated how antibiotic exposure alters EV responses in Staphylococcus aureus (SA)spiked blood and compared these findings with EV proteome profiles from bacteremia patients. In an in vitro model, whole blood from healthy donors was spiked with SA at a multiplicity of infection (MOI) of 0.001, treated with clinically relevant concentrations of piperacillin-tazobactam, vancomycin, or moxifloxacin, and plasma was subsequently isolated for EV analysis. EVs were isolated using the Miltenyi Pan EV Kit and analyzed by bead-based flow cytometry and high-resolution LC-MS/MS. In parallel, serum EVs from healthy controls (n = 6) and bacteremia patients (n = 12; 6 blood culture-positive and 6 culture-negative) were analyzed using the same workflow. Flow cytometry revealed increased levels of CMO⁺ CD45⁺ PanEV⁺ SA⁺ vesicles in SA-spiked samples, particularly following low-dose piperacillin-tazobactam and high-dose vancomycin treatment, despite minimal changes in vesicle size and total particle counts. Proteomic analysis of plasma EVs showed significant alterations in protein composition, including increased abundance of the SA-derived ribosomal protein rplU and host defense-associated proteins. Functional enrichment highlighted pathways related to neutrophil degranulation, vesicle-mediated transport, and antibacterial responses. In patient samples, serum EVs were enriched in acute-phase and immune-related proteins, including SERPINA1, SERPINA3, CRP, and SAA2, along with canonical EV markers such as CD81 and syntenin-1, irrespective of blood culture status. Antibiotic exposure and SA infection are associated with measurable changes in the human EV proteome, characterized by enrichment of immune and host defense-related proteins despite stable vesicle numbers. Similar EV-associated protein patterns were observed in both blood culture-positive and -negative patient samples, reflecting shared features of the systemic host response to infection and highlighting the potential of EV profiling to capture infection-associated biological signals.

<h4>Introduction</h4>Microenvironmental heterogeneity in cultured cells can compromise cell quality, reduce experimental reproducibility, and weaken the confidence of cell therapeutic efficacy. Although micropatterned cell cultures are more homogeneous, conventional micropatterning methods lack flexibility.<h4>Methods</h4>We develop a micropatterning technology by denaturing extracellular matrix (ECM) proteins in specific areas through heat inactivation using a high-speed laser via a light-responsive polymer layer. We have successfully seeded human induced pluripotent stem cells (hiPSCs) in flexible patterns and examine their neural induction in circular geometries of varying diameters.<h4>Results</h4>Size-dependent and cell-autonomous neural structures are formed on this substrate when hiPSCs differentiate into neural lineages in circles of different diameters. This self-organized pattern results from the mitotic orientation and localization of differentiating cells. Furthermore, teratogenic substances can modulate these patterns.<h4>Conclusions</h4>Laser-induced heat inactivation of ECM on culture substrates enables lithography-, hydrogel-, and PDMS-free micropatterning, facilitating on-demand regulation of cell-autonomous tissue formation, the effect of teratogenic substances, and precise tissue engineering in regenerative medicine.

West Nile virus (WNV) and Usutu virus (USUV) are neurotropic orthoflaviviruses of the <i>Flaviviridae</i> family, transmitted primarily by <i>Culex</i> mosquitoes and maintained in enzootic cycles involving birds. While WNV is a well-established human pathogen causing hundreds of neuroinvasive cases annually in Europe, USUV has emerged more recently, with fewer documented human infections but increasing evidence of neurovirulence. The viral nonstructural protein 1 (NS1) plays a central role in orthoflavivirus pathogenesis by modulating host immune responses, disrupting endothelial barrier integrity, and facilitating viral dissemination. However, the functional and biochemical properties of NS1 from WNV and USUV remain poorly characterized. We combined <i>in vitro</i>, <i>in vivo</i>, and clinical approaches to compare NS1 secretion, stability, and its impact on blood-brain barrier. Our results show that WNV NS1 is secreted at significantly higher levels, exhibits greater thermal stability, and disrupts brain endothelial barrier integrity <i>in vitro</i>. In contrast, USUV NS1 is secreted less efficiently, is slightly less stable, and does not compromise blood-brain barrier integrity, despite inducing distinct transcriptional responses in brain endothelial cells. In mice, WNV infection led to higher serum NS1 levels and stronger systemic inflammation than USUV. Clinically, WNV NS1 was detected mainly in patients with neurological symptoms, whereas USUV NS1 remained undetectable in all cases. Altogether, these findings reveal differential NS1 properties between these closely related viruses, with key implications for orthoflavivirus diagnosis and neurovirulence mechanisms.

Hand, foot, and mouth disease (HFMD) is primarily caused by Enterovirus 71 (EV71) and Coxsackievirus A16 (CA16), while excessive inflammatory responses induced by viral infection are the main cause of severe conditions. Hence, it is important to develop agents with both antiviral and anti-inflammatory activities for clinical treatment. In this study, we synthesized 20 compounds based on the hybridization of nonsteroidal anti-inflammatory drugs (NSAIDs) and organoselenium and found that YSN-1-167 exhibited significant anti-EV71 function using an EV71-GFP infection system. Mechanistic studies showed that this compound suppressed virus replication but not the binding and entry steps, and further target screening found that YSN-1-167 might suppress 3D polymerase function but not viral 2A and 3C proteases. The conservation of 3D polymerase between EV71 and CA16 prompted us to discover that YSN-1-167 can also significantly inhibit CA16 infection. As expected, this compound can reduce the levels of pro-inflammatory mediators, including IL-1β and COX-2, induced by EV71 infection. Furthermore, YSN-1-167 exhibited favorable safety and effective anti-EV71 and anti-inflammatory properties in neonatal mice. In conclusion, these results suggested that YSN-1-167 can be developed into a potential therapeutic strategy for HFMD induced by EV71 and CA16 via inhibiting virus replication and inflammatory response.

Chikungunya virus (CHIKV) infection remains a significant global health threat with no approved specific antiviral therapy. The nonstructural protein 2 protease (nsP2pro) is essential for viral replication and represents an attractive target for structure-based drug design. In this study, we employed a computational drug repurposing strategy to predict potential nsP2pro inhibitors from FDA-approved compound libraries. We performed molecular docking screens of 1820 unique compounds against the nsP2pro active site. The eight selected top-scoring compounds had docking scores better than that of the reference inhibitor pantinin-1. Subsequent molecular dynamics (MD) simulations revealed that, except for lomitapide, which dissociated from the pocket, the remaining leads formed dynamically stable complexes and modulated the catalytic environment in a chemotype-dependent manner. Small molecules expanded both the Cys478-His548 and Ser482-His548 distances, consistent with dyad perturbation, whereas GnRH-like peptides preserved a more compact geometry in the modeled noncovalent complexes, pointing to possible nonproductive occupation of the active-site region. Free energy perturbation calculations further provided a relative energetic ranking that supported the MD-derived mechanistic trends and placed six compounds ahead of pantinin-1 within the present data set. Among the small-molecule candidates, venetoclax and lapatinib emerge as computationally prioritized candidates for future biochemical and antiviral evaluation. Overall, this study provides mechanistic insight into noncovalent recognition of CHIKV nsP2pro and a framework for future experimental validation.

The double-stranded RNA-binding protein STAU2 plays an essential role in neural development and synaptic plasticity, assembling target mRNAs into ribonucleoprotein (RNP) granules to control their trafficking, localization, and local translation, though the detailed molecular mechanism remains elusive. Here, we show that STAU2 phase separates to form dynamic condensates in dendrites of hippocampal neurons, recruiting specific mRNAs to assemble mobile RNP granules that are transported distally along microtubules. These RNA-loaded STAU2 condensates undergo a liquid-to-gel transition, which stabilizes the encapsulated transcripts while repressing their translation. During neuron development, disrupting STAU2 condensation impairs RNP formation and compromises anterograde mRNA delivery to distal dendrites. Conversely, STAU2 overexpression promotes excessive coacervation, resulting in oversized RNP granules with reduced mobility, ultimately hindering dendritic elongation and promoting excessive branching. We further demonstrate that synaptic activity bidirectionally remodels STAU2 condensates in parallel with changes in local translation of bound mRNAs. Notably, aberrant STAU2 condensates emerge as a potential pathological feature in aggregation-prone neurodegenerative disorders. Collectively, our findings establish that STAU2 condensate safeguards the development of postmitotic neurons by orchestrating the dendritic transport and activity-dependent translation of its target mRNAs.

Neurological disorders such as Alzheimer's disease (AD), Parkinson's disease (PD), cerebral ischemia, anxiety and depression pose significant global public health challenges due to their high prevalence and complex pathological mechanisms. Current therapeutic strategies primarily offer symptomatic relief, with limited efficacy in mitigating disease progression. Neuroprotection involves interventions aimed at preserving neuronal structure and function through mechanisms such as reducing oxidative stress, modulating inflammation and inhibiting apoptosis, presenting a promising avenue for treating these conditions. Protocatechuic acid (PCA), a natural phenolic acid compound prevalent in a variety of foods and herbal medicines, has received considerable attention for its notable antioxidant, anti‑inflammatory and neuroprotective properties. The present study systematically reviews the neuroprotective effects and molecular mechanisms of PCA in various neurological disorders (including AD, PD and cerebral ischemia). The present review highlights the multi‑target mechanisms of PCA, which act by mitigating oxidative stress, neuroinflammation, mitochondrial dysfunction and apoptosis, while promoting neuronal regeneration. Furthermore, the present review integrates the body of evidence across neurological contexts to identify conserved protective pathways and discusses the translational potential of PCA, providing a foundation for its clinical application in treating neurological diseases.

BACKGROUND One-anastomosis gastric bypass (OAGB) is an effective bariatric surgery; however, there may be serious instances of metabolic and nutritional complications that may require complete reversal. Reversal of complete OAGB to near-normal physiology, especially with a background of sleeve gastrectomy, is extremely rare. This case report discusses the complications associated with reversal of OAGB. CASE REPORT Our patient was a 28-year-old woman who had a history of a sleeve gastrectomy procedure and later underwent a revisional OAGB procedure for poor weight loss/weight regain. She later developed weakness, chronic diarrhea, vomiting, severe protein-calorie malnutrition, refractory micronutrient deficiencies, and severe liver dysfunction with significant hepatocellular damage. She was admitted in unstable medical condition. After multidisciplinary evaluation, the patient underwent a laparoscopic reversal of OAGB to near-normal anatomy. The postoperative course was complicated by septic shock and multi-organ dysfunction requiring critical care interventions. With comprehensive multidisciplinary management, the patient gradually improved. After more than 8 months of follow-up, she demonstrated recovery in nutritional status, liver function, and functional capacity, despite the presence of a reduced gastric reservoir due to prior sleeve gastrectomy. CONCLUSIONS This case confirms that complete reversal of OAGB can be life-saving in selected patients with severe malnutrition and metabolic issues. Early identification of nutritional deficiencies and multidisciplinary care are critical to optimize outcomes in high-risk patients following bariatric surgery.

<h4>Background</h4>Porto-sinusoidal vascular disease (PSVD) is a complex, rare liver disease characterized by the absence of cirrhosis, with or without the presence of portal hypertension or histological lesions. Given the knowledge gaps in the mechanisms involved in this disease with unknown etiology, we used omics-based approaches to further elucidate the pathways affected by PSVD, facilitating improvements in the prognosis, diagnosis, and treatment options for these patients.<h4>Methods</h4>We applied gene set enrichment analysis (GSEA) and weighted gene coexpression network analysis (WGCNA) to identify pathways dysregulated in PSVD. Network construction and visualization were performed in Cytoscape to explore interconnectivity among enriched processes. Within key modules, candidate genes were prioritized by ranking approaches and cross-referenced with findings from previous studies.<h4>Results and conclusion</h4>In this study using both module eigengene correlation network analysis and GSEA, a novel coordinated dysregulation in PSVD was identified characterized by the simultaneous activation of immune and signaling pathways alongside the suppression of metabolic, ribosomal, and mitochondrial programs, highlighting a critical antagonistic interplay between these systems. Alterations in ribosomal proteins, ATP synthase subunits, and serpin family members highlight translational, bioenergetic, and anticoagulant dysfunction as core mechanisms. Together, these findings define PSVD as a disorder of integrated immune, vascular, and metabolic imbalance.

Monoallelic loss-of-function variants of GATA4 have been implicated in congenital heart defects, 46,XY differences of sex development, and congenital diaphragmatic hernia (CDH). However, there is no report of GATA4-variant positive patients who concomitantly exhibited these three features. Furthermore, the genotype-phenotype correlation of GATA4 abnormality remains unclear. Here, we report a 2-year-old boy who exhibited pulmonary valve stenosis, multiple atrial septal defects, hypospadias, bifid scrotum, bilateral inguinal hernia, and CDH. Endocrine evaluations of the patient were indicative of partial testicular dysgenesis. Exome sequencing identified a de novo 3-bp deletion in GATA4 that has not been reported previously. The variant eliminated one amino acid from the highly conserved five-threonine repeat in the second zinc finger domain (c.829_831del, p.T277del). The variant was assessed as deleterious by in silico analyses and was scored as "likely pathogenic" by the American College of Medical Genetics and Genomics guidelines. Our findings indicate that the five-threonine repeat in the second zinc finger domain contributes to GATA4 function. These results broaden the spectrum of GATA4 pathogenic variants and highlight the phenotypic diversity resulting from GATA4 abnormalities.

During aging, the progressive decline in neuronal function contributes to cognitive impairment and predisposes individuals to neurodegenerative disorders. This phenomenon has become increasingly prominent in modern society. Recent studies have found that post-translational modifications (PTMs) of proteins play a crucial role in the aging process, influencing the physiological functions and pathological changes in brain cells. This article reviews the variety and complexity of PTMs across brain cell types during aging, analyzes changes in specific modification types and sites, and reveals their impacts on neurophysiological and pathological states. This review also examines the current state of translational research, noting that most of the drugs under investigation lack specificity, and their potential toxic side effects remain undetermined. In light of these concerns, this review proposes therapeutic strategies to guide innovative interventions for aging-related diseases, aiming to facilitate further research and clinical applications in this field.

Hepatocellular carcinoma (HCC) is one of the most common malignancies worldwide, characterized by complex pathogenesis and immune escape mechanisms. The tumor immune microenvironment plays an increasingly recognized role in tumorigenesis, progression, and treatment. Receptor-interacting serine/threonine-protein kinase 2 (Ripk2) plays a pivotal role in inflammatory responses and immune regulation. This study aimed to investigate the role of Ripk2 in the HCC immune microenvironment, particularly its impact on macrophage function, using a macrophage Ripk2 knockout mouse model (Ripk2^CKO), single-cell sequencing analysis (scRNA-seq) and flow cytometry (FCM). We found that inhibiting or deleting Ripk2 altered the intratumoral microbiota within macrophages, significantly increasing the abundance of Streptomyces collinus (Sc). This alteration not only prompted a metabolic shift in macrophages but also significantly upregulated the transcription factor Myb expression, promoting Cxcl9 secretion, thereby enhancing CD8⁺ T cell infiltration, activity, and proliferation. Furthermore, Ripk2 was found to promote lactate production via the Pax5/Adpgk pathway, leading to CD8⁺ T cell dysfunction and forming an immunosuppressive feedback loop in HCC. Combining a Ripk2 inhibitor (GSK583) with a PD-1/PD-L1 inhibitor (BMS202) reduced the hepatotoxicity of BMS202 monotherapy and improved therapeutic efficacy. In conclusion, this study provides new insights into the role of Ripk2 in the HCC immune microenvironment and lays an experimental foundation for the development of Ripk2-targeted immunotherapy strategies.

Suppressed NMDA receptor (NMDAR) function contributes to multiple brain disorders, including schizophrenia, autism spectrum disorder (ASD), and NMDAR encephalitis. Previous attempts to restore NMDAR activity by increasing ambient glycine, a critical co-agonist, through GlyT1 inhibition have yielded mixed outcomes, partly due to GlyT1's extensive expression in essential brainstem regions. Slc6a20a, a glycine transporter widely expressed in cognition-relevant regions such as the cortex and hippocampus, offers a targeted alternative. Here we show that antisense oligonucleotide (ASO)-mediated Slc6a20a inhibition (Slc6a20a-ASO) normalizes ASD-related phenotypes in male Shank2- and Shank3-mutant mice, with model-dependent rescue profiles. Slc6a20a-ASO rescues NMDAR hypofunction and synaptic phospho-proteomic profiles in the prefrontal cortex. Furthermore, ASO targeting human SLC6A20 rescues suppressed NMDAR function in cortical organoids harboring SHANK2 or SHANK3 mutations. These findings underscore the potential and limitations of Slc6a20a/SLC6A20-ASO for treating disorders characterized by NMDAR hypofunction.

<h4>Background</h4>Intrahepatic cholangiocarcinoma (ICC) is a highly aggressive subtype of primary liver cancer with insidious onset, early metastasis, and poor prognosis. DNA damage response (DDR) dysfunction is linked to ICC tumorigenesis, progression, and drug resistance. However, the detailed mechanisms remain underexplored.<h4>Methods</h4>DDR activity was quantified by ssGSEA across 6 independent ICC cohorts. DDR risk model was analyzed using 10 machine learning algorithms to construct 101 combined models, and the optimal Lasso + RSF model was adopted by c-index. The core gene SFN was selected for experimental validation. Single‑cell RNA sequencing and spatial transcriptomics were used to explore cellular localization and tissue distribution of signature genes. In vitro assays were performed in ICC cell lines after SFN knockdown to assess cell proliferation and chemosensitivity.<h4>Results</h4>The DDR risk model showed stable prognostic performance in training and external validation cohorts. High‑risk patients exhibited poor prognosis, immunosuppressive microenvironment and reduced sensitivity to multiple chemotherapeutic agents. SFN was specifically enriched in malignant cells. Knockdown of SFN significantly suppressed cell proliferation and colony formation, enhanced sensitivity to cisplatin and gemcitabine, and increased γ‑H2AX expression indicating elevated DNA double‑strand breaks.<h4>Conclusions</h4>We constructed a robust DDR‑based prognostic model for ICC that enables accurate risk stratification and prediction of therapeutic response. SFN serves as a critical driver of chemoresistance and a potential therapeutic target. These results offer novel biomarkers and mechanistic insights to facilitate precision medicine for ICC patients.

Marfan syndrome (MS), Loeys-Dietz syndrome (LDS), Beals-Hecht syndrome (BHS), Ehlers-Danlos syndrome (EDS), and individuals with undifferentiated connective tissue disease (UCTD) exhibit phenotypic overlap, suggesting a likelihood of genotypic coexistence. Our objective was to evaluate genetic variants (GVs), encoding 174 genes related to aortopathies, cardiomyopathies, arrhythmias, structural heart disease, and hypercholesterolemia, and their relationship to clinical and cardiovascular damage in these syndromes. This was a prospective study in Mexican patients with MS, LDS, EDS, BHS, and UCTD. One hundred and seventy-four genes related to hereditary diseases were studied using next-generation sequencing targeting coding regions. Of the 136 patients, 25 were identified with the recurrent and coexisting GV of <i>MYBPC3</i>. In the MS group, in addition to the presence of GV in <i>FBN1</i>, eight patients had GV in <i>MYBPC3</i>, six in <i>FBN2</i>, and five in <i>COL3A1</i> and <i>COL5A1</i>. In the LDS group, in addition to GV in <i>TGFBR1</i>, <i>TGFBR2</i>, and <i>SMAD3</i>, four patients presented with GV in <i>MYBPC3</i> and two with <i>FBN2</i>. In the BHS group, in addition to <i>FBN2</i>, two patients had GV in MYBPC3 and one with <i>TGFBR2</i>. In the UCTD group, nine patients had GV in <i>MYBPC3</i> and two in <i>COL5A1</i> and <i>COL5A2</i>. All syndromes coexisted with GV in genes related to arrhythmias, sarcomeres, and hypercholesterolemia. In EDS, coexistence with several sarcomere proteins was found.

<b>Introduction</b>: Epilepsy is a chronic brain disorder causing recurrent seizures as a result of abnormal electrical brain activity. Approximately 70 million people worldwide suffer from it and need antiseizure treatment. The diagnosis and assessment of the effectiveness of the therapy are mainly based on symptoms described by a witness or a patient, which is a method of low reliability. The review aims to evaluate available markers of antiseizure drugs' efficacy, highlighting those that could support the proper course of treatment. <b>Results</b>: Comparing the available studies was very challenging. There were significant differences across the groups enrolled in the studies regarding the type of epilepsy included, the patient inclusion criteria, and the total number of patients. Moreover, most of the trials have been conducted on adults, not on children. <b>Conclusions</b>: Drug selection and their dosage are mainly based on the relationship between patients and their caregivers. Additionally, electroencephalography (EEG) and antiseizure medication (ASM) concentrations are assessed, but according to many studies, none of the applied methods are sufficient for the diagnostic and therapeutic process to be based on.

<h4>Background</h4>Hepatic steatosis is a common metabolic disorder for which accessible noninvasive assessment remains clinically relevant. This study aimed to evaluate the QCT-referenced performance of ultrasound attenuation (USAT) and an integrated model combining metabolic biomarkers for non-invasive categorization according to hepatic fat fraction in metabolic dysfunction-associated steatotic liver disease (MASLD), using quantitative computed tomography (QCT)-derived hepatic fat fraction categories as pragmatic imaging comparator labels.<h4>Methods</h4>A total of 172 participants were enrolled and categorized into hepatic fat fraction categories by QCT. USAT values, along with serum levels of ALT, AST, and ferritin, were collected. Three models were evaluated: USAT-only, laboratory-only, and an integrated USAT + laboratory model. Model performance was assessed using fold-separated internal validation, including nested five-fold stratified cross-validation with feature selection performed within training folds. Harrell's optimism-corrected bootstrap analysis was also performed as a supplementary internal validation method.<h4>Results</h4>USAT values increased significantly with hepatic fat fraction categories (P<0.001). In fold-separated internal validation, the USAT-only model achieved an AUC of 0.847 (95% CI, 0.780-0.902) for QCT-referenced detection of imaging-defined steatosis, while the laboratory-only model achieved an AUC of 0.753 (95% CI, 0.665-0.829). A fixed integrated model including USAT, ALT, and ferritin achieved an AUC of 0.845 (95% CI, 0.772-0.906), but did not significantly improve AUC compared with USAT alone. Multiclass categorization remained exploratory and limited, particularly for Category 2, which included only 31 participants and showed weak discrimination in the corrected random forest analysis (AUC=0.569 for the USAT + ALT + ferritin model). Subgroup analyses showed higher performance in females, younger participants, and those with higher BMI.<h4>Conclusion</h4>USAT shows promise as a noninvasive adjunct for QCT-referenced detection of imaging-defined hepatic steatosis in a single-center Chinese health-examination cohort, pending external validation. Adding ALT and ferritin may improve sensitivity and calibration, but did not significantly improve AUC over USAT alone. Because discrimination for the intermediate Category 2 group remained weak, the current model should not be considered reliable for full hepatic fat fraction categorization. Further validation in external, multicenter, and more diverse populations is required before any clinical use.

Adoptive T cell therapies can deliver curative responses for refractory patients with B cell malignancies, yet clinical activity in solid tumors remains inconsistent. Tumor-intrinsic barriers dominating this inconsistency include the immunosuppressive solid tumor microenvironment (TME) imposing chronic inhibitory cues to T cells and the scarcity of patient-shared and uniformly expressed tumor-restricted antigens for T cells to target. CRISPR-based forward genetics screens enable mapping of the functional genome regulating T cell anti-tumor activity. Here, we review recent insights from pooled CRISPR knockout screens in T cells to define convergent targets and pathways regulating T cell anti-tumor function and align the pharmacology of engineered T cells with sequential barriers they encounter within the TME. We additionally propose a framework for CRISPR screen-enabled target prioritization and present an example of how these principles can be applied to the functional enhancement of T cells through TIL (Tumor Infiltrating Lymphocyte) therapy, which utilizes a patient's personalized immune response against solid tumor antigens.

<h4>Introduction</h4>Multiple-stress is defined as the simultaneous or sequential exposure of plants to multiple abiotic constraints, which triggers regulatory programs that differ fundamentally from single-stress responses. In potato (<i>Solanum tuberosum</i> L.), drought, salinity, heat, and cold severely impair tuber development, yet the molecular architecture underlying resilience to combined stress remains unclear. We hypothesized that multi-stress conditions activate an integrated regulatory network linking tuber induction with stress-responsive metabolic and redox pathways.<h4>Methods</h4>RNA-seq profiling of microtuberization under combined osmotic, salinity, heat, and cold stress was performed. Differential expression analysis identified shared differentially expressed genes (DEGs). A subset of upregulated genes was used for protein-protein interaction (PPI) network construction. Comparative regulatory analyses were performed, and selected genes were validated by qPCR. Statistical analyses were conducted to assess differential expression and network enrichment.<h4>Results</h4>A total of 2,046 shared DEGs were identified, including 1,212 upregulated and 834 downregulated genes. A PPI network constructed from 1,475 unique upregulated genes revealed 317 highly interconnected components. Network analysis identified the StSP6A-FD tuberigen complex as a central regulatory hub integrating developmental signaling with phenylpropanoid metabolism, oxylipin biosynthesis, and redox regulation. Multiple components were associated with hydrogen sulfide (H₂S) signaling, suggesting redox-gasotransmitter integration.<h4>Discussion</h4>Comparative regulatory analysis revealed conservation of the ERF-NAC-MYB-bZIP transcription factor framework, along with expansion of stress-responsive modules. Collectively, these findings establish a mechanistic framework linking tuber induction with adaptive metabolic remodeling under multi-stress conditions.

This protocol describes sc‑rDSeq, a scalable, droplet‑based method for full‑length, strand‑specific total RNA sequencing at single‑cell resolution. The protocol uses a refined set of 220 ribosomal‑depleted sequences (rDS) primers that selectively exclude ribosomal RNA during initial reverse transcription, enabling capture of both polyadenylated and non‑polyadenylated RNAs such as histone RNAs, noncoding RNAs, and enhancer RNAs, without requiring costly post‑amplification depletion steps. This method is useful for researchers who would like to detect not only gene expression variations, but also alternative splicing events and single nucleotide variations in complex heterogenous cellular systems, providing a more complete view of cellular heterogeneity and regulatory programs that remain invisible to conventional polyadenylated‑only sequencing approaches. Compared with existing full‑length protocols, which are often limited by high reagent costs or reliance on complex multistep microfluidics, sc-rDSeq provides a simpler, single-step microfluidic workflow compatible with standard inDrops platforms, which may reduce experimental complexity and cost relative to existing full-length total-RNA methods. A key improvement is the 10-fold increase in unique molecular identifiers per cell relative to 3' end‑based methods, at a reported reagent cost of approximately $0.08 per cell, making deep total transcriptome analysis more accessible. The protocol includes three major parts: sc‑rDSeq barcode synthesis, single‑cell co‑encapsulation, and library construction.

Cutaneous hematologic neoplasms in children are relatively rare and encompass a wide range of lymphoproliferative and myeloproliferative disorders. This review explores and updates the classification, clinical presentation, diagnostic challenges, histopathology, and management of pediatric lymphomas, lymphoproliferations, and leukemias that may be seen in the skin. The most frequent of them are lymphomatoid papulosis (LyP) and mycosis fungoides (MF), and are discussed first, with a particular focus on differential diagnosis and overlaps with benign lesions-mainly pityriasis lichenoides-which raises questions regarding the delineation of these entities and their potential interconnection. It is important to underline that most cutaneous lymphoproliferations are indolent in children: primary cutaneous CD4+ small/medium T-cell lymphoproliferative disorder, subcutaneous panniculitis-like T-cell lymphoproliferation (non-associated with <i>HAVCR2</i> mutations), primary cutaneous marginal zone lymphoproliferative disorder, and EBV-related lymphoproliferative disorders. However, aggressive hematologic malignancies, although rarer, must not be missed; these are mostly leukemias (but not all forms) and blastic plasmacytoid dendritic cell neoplasm. We emphasize the importance of clinical-pathological correlation, with clonality studies playing a crucial role in some cases. Management strategies are briefly reviewed, ranging from skin-directed therapies like phototherapy and corticosteroids to systemic treatments for more aggressive forms of leukemia cutis and lymphomas.

<b>Background:</b> Low-dose aspirin (LDA) reduces preeclampsia (PE) risk by up to 40%, yet its molecular effects on chorion trophoblast cells (CTCs), a fetal membrane lineage at the feto-maternal interface, remain obscure. CTCs form a structural and immunoregulatory barrier whose dysfunction drives inflammation-associated membrane pathology in PE. Extracellular vesicles (EVs) released by CTCs may encode cellular stress and adaptation states, offering a molecular window into aspirin's timing-dependent effects on PE risk modification. <b>Methods:</b> Human CTCs were challenged with cigarette smoke extract (CSE) to model oxidative stress-driven PE pathology. Two paradigms were tested: (1) prophylactic aspirin (4 and 40 µg/mL) before and/or flanking the CSE, and (2) therapeutic aspirin after the CSE challenge. The EVs were isolated via ultracentrifugation and size-exclusion chromatography, characterized by nanoparticle tracking and immunoblotting, and profiled by quantitative mass spectrometry. A network pathway analysis and machine learning biomarker selection defined the EV-encoded molecular states. <b>Results:</b> The CTC-derived EVs from the CSE-exposed cells carried a PE-like proteomic signature marked by suppressed VEGF/ECM remodeling, activated TNF-p53 apoptotic signaling, and heightened inflammation. Prophylactic low-dose aspirin shifted the EV cargo toward an EV-encoded signature consistent with preserved angiogenic potential (enrichment of VEGFA, COL1A1, and MMP14) and predicted attenuation of apoptotic and NF-κB pathway activity by an Ingenuity Pathway Analysis. High-dose aspirin produced broad transcriptional suppression without an accompanying pro-angiogenic EV signature. Therapeutic (post-injury) aspirin partially attenuated the injury-associated EV cargo but did not restore the angiogenic EV signature. An exploratory machine learning analysis of EV proteomes identified a candidate prophylactic biomarker panel anchored by HSPA8, SERPINF2, COL4A1, and PLOD1, mapped to the predicted angiogenic recovery and redox-balance pathways. These EV cargo readouts represent the predicted molecular states and require functional validation before clinical interpretation. <b>Conclusions:</b> The CTC-derived EV proteomic signatures capture the dose- and timing-dependent aspirin effects in this in vitro CTC model, positioning the chorion as a candidate pharmacological "secondary responder" favoring cellular resilience over classical anti-inflammatory suppression. As an exploratory hypothesis-generating study, EV-based molecular profiling could provide a foundation for future investigations aimed at stratifying aspirin responders from non-responders, although clinical validation in maternal plasma cohorts will be required before any translational application.

Adolescent idiopathic scoliosis (AIS), the spontaneous development of a lateral spine curvature during puberty, is the most common pediatric spine disorder, affecting ∼3% of children worldwide. As the underlying etiology remains unclear, AIS is treated purely symptomatically, initially by bracing and ultimately by highly invasive, costly surgeries. Genome-wide association studies (GWAS) have identified numerous risk loci in non-coding genomic regions, making it difficult to link them to a biological function. To address this, we performed a multi-tissue investigation to connect genetic risk to tissue-specific molecular pathology. We conducted RNA sequencing on the primary tissues implicated in AIS, paraspinal muscle and spinal cartilage, from patients and unaffected controls. In paraspinal muscle, we identified differentially expressed genes (DEGs) enriched for pathways related to muscle structure, myogenesis, and metabolism. Key upregulated genes include the transcription factor EGR1 and structural components such as MYH1 . In spinal cartilage, we found enrichment of genes related to TGFβ and FoxO signaling, as well as metabolic pathways. Notably, genes crucial for chondrocyte differentiation (e.g. SOX5 and SOX6 ) were significantly downregulated. We then examined genes at known GWAS loci and found that several risk-associated genes were differentially expressed in one or both tissues. To investigate the function of non-coding variants at these loci, we identified and validated several enhancer elements harboring AIS risk SNPs at the BCL2, ADGRG6, BNC2, and FTO loci. We reveal distinct pathological signatures in muscle and cartilage and lay the foundation for connecting non-coding genetic risk to the dysregulation of key developmental and structural pathways.

<h4>Background</h4>Curcumin is the major component of turmeric, which has an anticancer property in multiple cancers, including hepatocellular carcinoma (HCC). However, the mechanisms are still largely unclear. This research aims to assess the pharmacological function of curcumin and explore the potential microRNA (miRNA)-mRNA regulatory mechanism in curcumin-mediated HCC progression.<h4>Materials and methods</h4>Hep3B and Huh-7 cells were used for <i>in vitro</i> experiments. Cells were exposed to various doses of curcumin, and transfection was conducted using Lipofectamine 2000. Cell proliferation, migration, and invasion were examined using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide or transwell assay. The abundances of miR-21-5p and sex-determining region Y-related high-mobility group box 6 (<i>SOX6</i>) were examined using quantitative real-time polymerase chain reaction or Western blot. The relationship between miR-21-5p and <i>SOX6</i> was analyzed through luciferase reporter analysis.<h4>Results</h4>Curcumin repressed HCC cell proliferation, migration, and invasion. miR-21-5p level was decreased in curcumin-treated cells, and miR-21-5p overexpression reversed curcumin-mediated inhibition of HCC progression. <i>SOX6</i> was targeted through miR-21-5p, and <i>SOX6</i> restoration attenuated miR-21-5p-induced promotion of HCC progression. Moreover, curcumin exposure increased <i>SOX6</i> expression through regulating miR-21-5p, and knockdown of <i>SOX6</i> overturned curcumin-modulated suppression of HCC progression.<h4>Conclusions</h4>Curcumin repressed proliferation, migration, and invasion of HCC cells by regulating miR-21-5p and <i>SOX6</i>, indicating the promisingly pharmacological effect of curcumin in HCC.

Mitochondrial dysfunction is a core pathogenic mechanism underlying a broad spectrum of neurodegenerative disorders, from Alzheimer's and Parkinson's diseases to inherited optic neuropathies and mitochondrial ataxias. This review provides a comprehensive analysis of how defects in mitochondrial and nuclear DNA converge to disrupt oxidative phosphorylation, mitochondrial dynamics, calcium homeostasis, and quality control pathways, leading to energy depletion, oxidative stress, and neuronal degeneration across multiple disease contexts. Building on this mechanistic foundation, we examine how these shared pathogenic principles manifest distinctly in major neurodegenerative diseases, while also discussing representative mitochondrial optic neuropathies as tractable disease models that have yielded critical mechanistic and therapeutic insights. We further review recent advances in diagnostic technologies that enhance our ability to detect and stratify mitochondrial pathologies for therapeutic intervention. On the therapeutic front, we provide a comprehensive evaluation of the rapidly evolving landscape, analyzing strategies ranging from metabolic modulators and antioxidants to pioneering gene-targeted therapies, organelle replacement approaches, and emerging epitranscriptomic interventions. Finally, we identify persistent challenges in clinical translation and outline pivotal future directions essential for developing effective, mechanism-informed combination therapies against mitochondrial dysfunction in neurodegeneration.

Cholesterol metabolites are abundant in the central nervous system (CNS) that regulate cell membrane fluidity, signal transduction, and inter- and intracellular vesicular transport, as well as cell proliferation/cell death or migration. Brain cholesterol synthesis and metabolism are tightly coupled to the functional homeostasis of neurons, glial cells or microglia, and dysregulation of these processes has been strongly implicated in neurodegenerative diseases such as Alzheimer's disease (AD), Parkinson's disease (PD), and Huntington's disease (HD). This review provides a comprehensive overview of how cholesterol synthesis, esterification, efflux, uptake, and oxidation affect the CNS function, highlighting the function of key enzymes or metabolites in distinct brain cell types during neurodegeneration. Based on single-cell/nucleus RNA sequencing data from the brains of AD, PD, and HD patients, we summarize cell-type-specific genes in cholesterol metabolism pathways, shedding new light to understand cellular heterogeneity. The role of cholesterol-derived neurosteroids in neurodegenerative diseases is also discussed. Furthermore, how cholesterol metabolites modulate the formation, aggregation, and degradation of amyloid-β (Aβ), α-synuclein and huntingtin, as well as Tau protein phosphorylation are outlined. Finally, future research directions are proposed that aim to understand neurodegenerative diseases with new angle.

<h4>Background</h4>It is generally considered that eukaryotic translation initiation prominently occurs from the first AUG codon by ribosomal scanning from the 5-cap end of each mRNA. In order to identify cryptic internal translation initiation sites defined by alternative AUG codons on a proteome-wide scale, we generate a customized amino acid sequence database which contain differential AUG-guided tryptic peptide fragments computationally predicted from well-curated Swiss-Prot human protein reference sequences and applied it for high-resolution mass spectrometry-based proteomic analysis.<h4>Results</h4>The ultra-deep proteomic detection based on the real-time search platform on Orbitrap Eclipse Tribrid mass spectrometry system leads to identification of not only more than 26,000 unique peptides from already annotated human protein coding sequences but also 794 novel peptide fragments defined by alternative downstream translation initiation in human cancer cells. Very notably, Tandem Mass Tag-based multiplex quantitative analysis of patient-derived glioblastoma initiating cells uncovers epidermal growth factor-dependent translational regulation on a wide range of differential AUG-guided non-canonical proteoforms as well as cancer-related transcription factors and cell cycle/cell division regulators in a cell-type specific manner.<h4>Conclusions</h4>Our study provides the first proteome-wide evidence of downstream AUG-guided cryptic translation initiation dynamics in human cancer cells.

<h4>Background</h4>Mutations in antithrombin (<i>SERPINC1</i>) lead to the hereditary antithrombin deficiency. Conventional therapies for hereditary antithrombin deficiency are prophylactic or on-demand oral anticoagulants, which have poor compliance and side effects. This study explored the therapeutic efficacy of adeno-associated virus serotype 8 (AAV8)-mediated human <i>SERPINC1</i> gene (AAV8-hSERPINC1) in AT (antithrombin)-deficiency mice.<h4>Methods</h4>AAV8-hSERPINC1 carrying luciferase was injected into AT^+/- mice via tail vein injection at low, medium, and high doses. The biodistribution and expression of the carrier were visualized by in vivo bioimaging technology. Plasma AT levels were serially monitored by ELISA, and an inferior vena cava model was established to evaluate thrombotic propensity. Safety was evaluated by monitoring hepatic, renal, and cardiac function parameters and employing flow cytometry.<h4>Results</h4>A dose-dependent increase in AT expression was observed in AT^+/- mice after AAV8-hSERPINC1 injection. Compared with the untreated mice, medium-dose treatment restored plasma AT activity and antigen in AT^+/- mice to normal levels by week 8, with maintenance within the normal reference range for 40 weeks. In the venous thrombosis model, the rate of thrombosis in mice treated with medium-dose AAV8-hSERPINC1, rivaroxaban, low-molecular-weight-heparin, and wild-type mice were 60%, 60%, 70% and 60%, respectively. After AAV injection, transient elevations in hepatic transaminases and cytokines were observed in mice during a short-term period.<h4>Conclusions</h4>Our study demonstrates that AAV8-hSERPINC1 gene delivery resulted in durable AT expression, sustained blood hypercoagulation state correction, thereby rescuing thrombophilia in AT-deficient male mice. These data support the long-term efficacy and safety of AAV gene therapy for hereditary antithrombin deficiency.

<h4>Background</h4>Current bipolar disorder (BD) therapies suffer from limited efficacy and adverse effects, necessitating mechanistically grounded targets.<h4>Methods</h4>We integrated BD genome-wide association study data (158,036 cases; 2,796,499 controls) with brain proteomics (ROSMAP and Banner dorsolateral prefrontal cortex, <i>n</i> = 376 and 152) to perform proteome-wide association studies (PWAS). Bayesian colocalization and summary-data-based Mendelian randomization (SMR) prioritized causal genes. Cell-type-specific transcriptomics validated dysregulation in iPSC-derived neurons, astrocytes, and postmortem hippocampus/prefrontal cortex. Weighted gene co-expression networks (WGCNAs), functional enrichment, and molecular docking assessed functional pathways and druggability.<h4>Results</h4>PWAS identified eight BD-associated genes (false discovery rate < 0.05), with <i>DOC2A</i> emerging as the top candidate. Colocalization (<i>H₄</i> > 0.8) and SMR supported a causal association of <i>DOC2A</i> with BD, with no pleiotropy (heterogeneity in dependent instruments <i>P</i> > 0.01); <i>DOC2A</i> expression decreased in BD across neurons (<i>P</i> = 4.26 × 10^-2), astrocytes (<i>P</i> = 2.09 × 10^-2), hippocampus (<i>P</i> = 9.80 × 10^-3, <i>t</i> = -2.738), and prefrontal cortex (<i>P</i> = 1.44 × 10^-2, <i>t</i> = -2.580); WGCNA positioned <i>DOC2A</i> as a key regulator (module membership/gene significance <i>P</i> < 0.05) of co-expression networks enriched for BD-associated processes including neurotransmitter secretion and postsynaptic actin cytoskeleton organization (<i>P</i> < 0.05); molecular docking revealed favorable-affinity binding (ΔG < -4 kcal/mol) between DOC2A and BD-related drugs and neuroprotective compounds.<h4>Conclusions</h4>Our convergent multi-omics framework highlights <i>DOC2A</i> dysregulation as a key contributor to synaptic dysfunction in BD and nominates it as a promising therapeutic target. The demonstrated interaction with existing neuroactive compounds provides immediate translational avenues.

Maintaining a functional proteome is essential for cellular health and organismal longevity. Disruption of proteostasis is a hallmark of aging and a central driver of diverse pathologies, including neurodegeneration, cancer, and metabolic disease. The ubiquitin-proteasome system (UPS) and autophagy represent the two principal degradative pathways safeguarding proteome integrity, particularly under conditions of stress. While historically viewed as mechanistically distinct, it is now clear that UPS and autophagy operate as an interconnected and adaptive network. This Perspective discusses three core principles that govern their coordination: (1) a shared molecular language of ubiquitin signals and shuttle proteins that determines cargo routing; (2) spatial compartmentalization through organelle-specific quality control modules and phase-separated degradation hubs; and (3) temporal regulation by stress-responsive signaling pathways that reprogram proteolytic output. Understanding this dynamic partnership not only reveals fundamental organizing principles of cellular homeostasis but also identifies new therapeutic nodes for diseases driven by proteostasis collapse.

<h4>Background</h4>Lung adenocarcinoma (LUAD), the predominant histological subtype of non-small cell lung cancer, remains a leading cause of cancer-related mortality worldwide. The RAS signaling pathway plays a critical role in LUAD pathogenesis; however, the heterogeneity of RAS pathway activity and its clinical implications remain poorly understood. This study aimed to characterize RAS pathway activity subtypes and develop a robust prognostic model for LUAD patients.<h4>Methods</h4>Transcriptomic data from 624 LUAD patients (GEO datasets: GSE31210 and GSE72094) were analyzed as the training cohort, with TCGA-LUAD as validation. Consensus clustering stratified patients based on 238 RAS pathway-related genes. Candidate genes were identified through differential expression analysis and WGCNA. Machine learning algorithms (LASSO, Random Forest, SHAP) were applied to construct a prognostic risk model. Comprehensive analyses including GSEA, CIBERSORTx-based immune infiltration, ESTIMATE scoring, and drug sensitivity prediction were performed.<h4>Results</h4>The study identified two distinct RAS pathway activity subtypes among LUAD patients. A three-gene prognostic signature (MAPK10, PLA2G12B, SHC3) was established, with the risk score serving as an independent prognostic indicator. Risk score was an independent prognostic factor. Immune landscape analysis demonstrated that high- and low-risk patients showed different expression levels of immune cells. All signature genes correlated positively with resting mast cells, while MAPK10 and PLA2G12B negatively correlated with activated CD4 + memory T cells. GSEA revealed high-risk tumors enriched in DNA replication, cell cycle, and base excision repair, whereas low-risk tumors favored drug and retinol metabolism pathways. Low-risk patients exhibited lower TIDE and exclusion scores, indicating better immunotherapy response potential. Eight therapeutic compounds (genistein, metformin, quercetin, JNK-9 L) demonstrated favorable binding to signature genes.<h4>Conclusion</h4>This study established a comprehensive landscape of RAS pathway activity subtypes in LUAD, identifying MAPK10, PLA2G12B, and SHC3 as novel prognostic biomarkers with significant associations with immune microenvironment remodeling, providing new insights for personalized treatment strategies and therapeutic target development.

Selenium (Se) is an essential trace element whose physiological functions in redox regulation, immunity, and metabolism are primarily executed by 25 distinct human selenoproteins. While systemic Se status is maintained through a tightly regulated hepatic distribution hierarchy and receptor-mediated transport to vital organs, recent research has significantly advanced our understanding of the specific roles of individual selenoproteins. This review focuses on the thioredoxin-like family, comprising SELENOT, SELENOV, SELENOW, and SELENOH, which share a conserved Rdx fold but perform diverse, tissue-specific functions. SELENOT has been implicated in the development and function of catecholaminergic circuits and may contribute to the regulation of calcium homeostasis, while SELENOV appears to play a role in modulating energy balance, adiposity, and hepatic responses to stress resistance. Emerging evidence suggests that SELENOW may play roles in muscle maintenance, bone remodeling, and inflammation resolution, with potential involvement in neuroprotection via regulation of tau homeostasis. Finally, the nuclear-localized SELENOH preserves genomic stability and is involved in cell cycle progression. By integrating findings from molecular pathways and physiological data from knockout mouse models, this review highlights the roles of these four proteins in supporting cellular viability and physiological homeostasis under conditions of oxidative and metabolic stress.

<h4>Background</h4>Venous thromboembolism (VTE) is a major contributor to morbidity and mortality in severe trauma (ST) patients. Platelet count (PLT), fibrinogen level (FIB), and D-dimer level are important coagulation indicators; however, few studies have described their trajectory in ST and assessed predictive value for VTE in clinical and nursing practice.<h4>Objective</h4>To describe the trajectories of PLT, FIB, and D-dimer on days 1, 3, 5, and 7 after ST, and to evaluate their predictive value for post-traumatic VTE.<h4>Methods</h4>Clinical data, including PLT, FIB, and D-dimer, were retrospectively collected from 184 ST patients. Univariate analysis and multivariate logistic regression analysis were used to identify independent risk factors for intramuscular venous thrombosis (IMVT), deep vein thrombosis (DVT), and pulmonary embolism (PE) in ST patients. Receiver operating characteristic (ROC) curve analysis was performed to evaluate the predictive performance of these independent risk factors.<h4>Results</h4>PLT showed a decreasing trend from day 1 to day 3 after trauma, followed by an increasing trend from day 3 to day 7. FIB dropped sharply to the lowest value on day 1 after trauma and then gradually increased. D-dimer surged sharply to the peak on day 1 after trauma, decreased gradually in the first 3 days, and then increased slowly from day 3 to day 7. Multivariate logistic regression analysis identified D-dimer level on day 5 as an independent risk factor for both IMVT (OR=1.133, 95% CI: 1.032-1.243; P=0.009; AUC=0.561) and PE (OR=1.085, 95% CI: 1.016-1.160; P=0.015; AUC=0.721). PLT and FIB were not identified as independent risk factors for IMVT, DVT, or PE.<h4>Conclusions</h4>This study demonstrated that while the dynamic changes in PLT, FIB, and D-dimer can reflect the transition of coagulation status after ST, their value as independent VTE predictive tools is limited. In clinical and nursing practice, it is crucial to determine the timing of coagulation status transition after trauma and explore the optimal timing for pharmacological prophylaxis.

<h4>Background</h4>Glycemic control involves methods for measuring, evaluating, and correcting blood glucose levels to maintain glucose homeostasis. It is essential to reduce hyperglycemia-related complications while minimizing the risk of hypoglycemia, and registered nurses (RNs) play a key role in blood glucose monitoring and insulin therapy management in the intensive care unit (ICU).​.<h4>Objectives</h4>To provide a thorough synthesis of nursing perspectives on glycemic control practices for critically ill adult patients admitted to the ICU.<h4>Methods</h4>We conducted a narrative review of the literature using PubMed, Scopus, CINAHL, and Google Scholar, complemented by manual searches of relevant scientific society websites, textbooks, and reference lists.<h4>Results</h4>Available evidence suggests initiating glycemic control for persistent hyperglycemia at ≥180 mg/dL (10 mmol/L) and titrating therapy to an individualized target based on patient evaluation. Arterial blood samples should be prioritized for glucose measurement when available, followed by venous and capillary sampling, individualizing the monitoring frequency. Continuous intravenous insulin infusion is generally recommended, guided by a standardized protocol specifying treatment thresholds, target ranges, insulin infusion adjustments, and monitoring frequency.<h4>Conclusions</h4>This narrative review summarizes nursing perspectives on glycemic control in critically ill adult ICU patients, including glucose measurement, individualized monitoring frequency, and protocol-based intravenous insulin infusion management. Evidence gaps remain regarding the optimal application of these approaches, and nursing research should focus on both patient-centered and organizational outcomes.

Scoliosis affects 2-3% of people, often developing during and after adolescence, and currently has a lifetime chance of surgical intervention of ~0.1% in high income countries. Understanding of causal genetic and environmental factors is improving, with mechanical feedback interactions between the neuromuscular and skeletal systems thought to be important. While examining mechanosignalling in the zebrafish musculoskeletal system, we observed transient expression of yap1 mRNA in precursor cells of muscle and notochord and wwtr1 mRNA accumulation in differentiated muscle. Yap1 and Wwtr1/Taz are transcriptional coactivators that mediate Hippo pathway signalling, often in response to mechanosignals. Loss of function mutation of either gene alone transiently altered early larval motility and reduced survival to adulthood, but mutation of yap1 specifically diminished overall growth without an obvious histological muscle defect. Yap1 mutants had a temperature-sensitive phenotype of oedema in cardiac and other tissues, which could be rescued by rearing at low temperature. Rescued yap1 mutants showed focal defects in hypochordal col8a1a mRNA expression at 1-2 days post-fertilisation (dpf), an early motility defect at 5 dpf and subsequently developed a fully penetrant vertebral dysmorphology, reflected by a decrease in posterior vertebral height. Thereafter, frank kyphoscoliosis accompanied by additional vertebral defects developed in around a third of the surviving yap1 mutants and was first detected at 11 dpf. Thus, the mild initial vertebral defect can, in a predisposing genetic or environmental background, gradually develop into full kyphoscoliosis through a positive feedback mechanism, analogous to the Hueter-Volkmann 'Law'. Although the cell type/s of cell autonomous yap1 action remain unclear, we hypothesise that Yap1 mechanosensation mediates feedback between bone, muscle and tendon to restrain vertebral overgrowth and protect against the development of kyphoscoliosis.

<h4>Objective</h4>Regular exercise reduces the risk of obesity, but the impact of endurance exercise on skeletal muscle metabolic processes remains incompletely understood. This study investigated the effects of endurance exercise on the transcriptome and metabolome of skeletal muscle in obese mice.<h4>Method</h4>Transcriptomic and metabolomic profiling of skeletal muscle samples was performed across experimental groups to identify differentially expressed genes and differential metabolites. Functional enrichment of DEGs and metabolites was analyzed using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses. Integrated pathway-level analysis was conducted using iPath 3.0.<h4>Results</h4>Integrated transcriptomic and metabolomic analysis indicated that endurance exercise altered lipid metabolism, amino acid metabolism, and nucleotide metabolism pathways in the skeletal muscle of obese mice. Specifically, endurance exercise downregulated the expression of lipid metabolism-related gene Acot2, amino acid metabolism-related gene Nos2, and carbohydrate metabolism-related genes Itpkb and Inpp4b. Furthermore, endurance exercise decreased the levels of palmitic acid, arachidonate, phosphatidylcholine, acetylcholine, hypoxanthine, urea, and acetyl-CoA.<h4>Conclusions</h4>Endurance exercise positively impacts skeletal muscle metabolic health in obese mice through coordinated regulation of multiple metabolic pathways and reduction of specific metabolites.

<h4>Background</h4>Many patients with cancer benefit little from immune checkpoint blockade (ICB), a major obstacle to immunotherapy for decades. Finding alternative immune checkpoints that control CD8⁺ T-cell exhaustion is urgent if we are to improve the efficacy of immunotherapies, particularly in microsatellite stable (MSS) colorectal cancer (CRC) that is resistant to ICB.<h4>Methods</h4>Spatial proximity is essential for suppressive ligand-receptor signaling. Here, we mapped the spatial tumor microenvironment of patients with MSS CRC at single-cell resolution and analyzed the cells interacting with exhausted CD8⁺ T cells to identify immune checkpoint ligand-receptor pairs. To investigate the function of this previously unrecognized immune checkpoint, we performed validation studies spanning cellular experiments, mouse models, and clinical patient samples.<h4>Results</h4>We found that a subset of MSS CRC exhibits substantial CD8⁺ T-cell infiltration, but their function is suppressed. We identified CLEC2B (ligand)-KLRB1 (receptor) as a novel inhibiting ligand-receptor pair for CD8⁺ T cells. KLRB1 acts as an immune checkpoint receptor, increasing CD8⁺ T-cell exhaustion and facilitating immune escape in various human cancers. Binding of CLEC2B to KLRB1 initiates immunosuppressive signaling in CD8⁺ T cells. Clinically, CLEC2B-KLRB1 expression correlates positively with cancer progression and poor response to ICB, demonstrating that KLRB1⁺ CD8⁺ T cells are a key marker of the poorly responsive ICB subtype. Furthermore, blocking CLEC2B-KLRB1 signaling with antibodies enhances the antitumor function of CD8⁺ T cells, providing a potential immunotherapy target for ICB non-responders.<h4>Conclusions</h4>Our study revealed CLEC2B-KLRB1 as a previously unrecognized immune checkpoint axis that drives T-cell exhaustion specifically in ICB poor responsive MSS CRC. Blockade of KLRB1 with a therapeutic antibody reinvigorated CD8⁺ T-cell antitumor immunity, positioning this axis as a promising target for enhancing immunotherapy efficiency in malignancies, including CRC and other ICB-resistant cancers.

<h4>Background</h4>Chronic pancreatitis (CP) is a progressive fibroinflammatory disorder with persistent immune activation and limited therapeutic options. While physical activity (PA) benefits many chronic diseases, it is often presumed neutral or potentially harmful in CP.<h4>Objective</h4>To assess whether PA protects against CP and defines the underlying mechanisms.<h4>Design</h4>We analysed the association between PA and CP risk in the UK Biobank cohort (>500 000 participants) and validated findings in an independent clinical cohort. In mice, experimental CP was induced and the effects of exercise interventions on pancreatic injury, fibrosis and immune responses were evaluated via histopathology, immunohistochemistry, flow cytometry, bulk and single-cell RNA-sequencing and proteomics.<h4>Results</h4>In the UK Biobank, regular PA was independently associated with a lower risk of CP. This association was consistent across alcohol intake strata and disease subtypes. Consistently, physically active patients with CP exhibited milder clinical manifestations. In mice, exercise interventions, including both preconditioning and postdisease initiation, attenuated pancreatic injury, fibrosis and ferroptosis, with resistance exercise providing greater protection. Mechanistically, skeletal muscle-derived extracellular vesicles (EVs) induced by PA accumulated within inflamed pancreata and dampened mitochondrial DNA-driven innate immune activation while promoting inflammation-resolving states, at least in part through modulation of myeloid stimulator of interferon genes (STING) signalling. Importantly, inhibition of EV release partially attenuates these protective effects. Proteomic profiling identified PRDX6 as a muscle-derived vesicular factor that inhibits ferroptosis and, by binding to the zinc-thumb motif of cyclic GMP-AMP synthase, contributes to suppression of STING activation and inflammatory damage.<h4>Conclusion</h4>PA restrains CP progression by reprogramming pancreatic immune responses and ferroptosis pathways.

Pain-related conditions are the leading cause of disability worldwide. Existing GWAS for chronic pain have mainly focused on individual pain-related disorders, which may not optimally capture the phenotype. Here, we define chronic pain based on prescription analgesic use ( ≥ 90 days) in two large biobanks (UK Biobank and FinnGen). GWAS meta-analyses of 11 prescription-based pain phenotypes identify 140 associations with chronic pain, including 78 novel (e.g. ARPP21, CNTNAP2) and 62 previously reported (e.g. SLC39A8, DCC, TRPM8) associations. Integrating these genetic associations with functional data including transcriptome-wide association studies, cell-type and pathway enrichment, and gene enrichment in mouse phenotypes identifies potential mechanisms involved in chronic pain, implicating oligodendrocyte differentiation, neuronal guidance, endolysosomal function and post-synaptic endosome recycling. Our study showcases how the use of prescription data to identify and characterize pain can provide insights into pain genetics and its underlying biology.

Estrogen receptor-positive breast cancer represents a significant proportion of breast cancer brain metastasis but remains understudied. Here we show that FGFR1-amplification, a well-established driver of estrogen receptor-positive breast cancer endocrine resistance, promotes estrogen receptor-positive breast cancer brain metastatic colonization in young and aged female mice, through both canonical FGF2/FGFR1 signaling and non-canonical NCAM1/FGFR1 interactions. Astrocytic FGF2-mediated paracrine activation of FGFR1 promotes breast cancer brain metastasis in estrogen-treated young mice, but FGF2 levels and signaling decrease in the brain with aging and estrogen-depletion. Neuronal and astrocytic NCAM1, which remain unchanged in young and aged brains, promote adhesion to neurons, migration, and growth of estrogen receptor-positive cells, suggesting that interactions with astrocytes and neurons facilitate early estrogen receptor-positive breast cancer brain metastasis colonization through FGFR1. Importantly, FDA-approved FGFR inhibitors effectively block early colonization but not late-stage brain metastases, suggesting prevention of FGFR1+ brain metastases as a window of opportunity for FGFR1 inhibitors.

<h4>Background</h4>Osteoporosis, characterized by reduced bone mass and increased fracture risk, underscores the urgent need for novel anabolic therapies. G protein-coupled receptors (GPCRs) are major drug targets, but the functions of adhesion GPCRs (aGPCRs) in bone remain largely unexplored. G-protein coupled receptor 125 (Gpr125) is an orphan aGPCR, and its role in osteoblast-mediated bone formation is entirely unknown.<h4>Methods</h4>We employed lentiviral-mediated knockdown and overexpression of Gpr125 in primary mouse calvarial osteoblasts and bone marrow stromal cells (BMSCs). Osteogenic and adipogenic differentiation were assessed by staining and marker analysis. Transcriptomic profiling (RNA-seq) and pathway analysis were used to identify downstream mechanisms, validated by rescue experiments with Gper1 overexpression, PI3K/AKT inhibitors (LY294002), and Wnt/β-catenin activation (Wnt3a CM). The osteogenic role of Gpr125 was tested in ovariectomized and aged mouse osteoporosis models via osteoblast-targeted adeno-associated virus (AAV) delivery.<h4>Results</h4>Gpr125 was highly expressed in osteoblasts, peaking during differentiation. Its knockdown severely impaired osteogenesis while promoting adipogenesis in vitro. Conversely, its overexpression enhanced bone formation. RNA-seq identified G protein-coupled estrogen receptor 1 (Gper1) as the key downstream target. We defined a novel signaling axis where Gpr125 upregulates Gper1, which activates PI3K/AKT signaling, leading to β-catenin stabilization and osteogenic transcription. Rescue experiments established a strict hierarchy: Gper1 overexpression fully rescued the osteogenic defect caused by Gpr125 loss, but not Gpr125 expression itself. PI3K inhibition blocked Gpr125-induced β-catenin activation and osteogenesis. β-catenin activation partially rescued osteogenesis but failed to restore upstream signaling. Critically, osteoblast-specific Gpr125 overexpression in vivo effectively ameliorated bone loss and reduced marrow adiposity in both postmenopausal and senile osteoporosis mouse models.<h4>Conclusions</h4>Our study unveils a complete Gpr125-Gper1-PI3K/AKT-β-catenin signaling axis essential for osteoblast differentiation and bone formation. This work identifies the aGPCR Gpr125 as a novel positive regulator of bone anabolism and proposes the Gpr125-Gper1 axis as a promising therapeutic target for developing new treatments against osteoporosis.

<h4>Background</h4>Patients with lung cancer brain metastases can benefit from immune checkpoint inhibitors (ICI). However, intracranial responses are often limited and not always concordant with activity seen in extracranial disease. Defects in IFNγ signaling and HLA class-I antigen presentation machinery (APM) on malignant cells can drive immune evasion and ICI resistance, and have traditionally been viewed as interdependent. The possible role of these alterations in non-small cell lung cancer (NSCLC) brain progression remains poorly understood.<h4>Methods</h4>Using multiplex quantitative immunofluorescence, we measured and spatially mapped IFNγ signaling markers (pSTAT1 and IRF1) and multiple HLA class-I APM components (β2M, PSMB8, PSMB9, PSMB10, TAP1, TAP2, Tapasin, Calreticulin, and ERp57) in cancer cells and neighboring non-malignant stromal cells from two patient cohorts, including primary tumors, intra- and extra-cranial NSCLC metastases. We also studied tumor-infiltrating lymphocyte (TILs) subpopulations in the cohorts, performed whole transcriptomic analysis of parental human NSCLC H2030 cells and their brain metastatic counterpart H2030-BrM3, and expanded the results using spatial transcriptomics of human tumors.<h4>Results</h4>We found comparable levels of IFNγ signaling markers in primary and metastatic lesions and marked downregulation of multiple APM components in metastases, some of which were restricted to the brain. Downregulation of HLA class-I APM components was associated with reduced effector TILs and worse survival. Analysis of human parental H2030 and brain metastatic H2030-BrM3 lung adenocarcinoma cells showed comparable signaling responses after IFNγ stimulation and reduced HLA class-I APM markers in metastatic cells. Transcriptomic analysis of primary/metastatic cells and human tumors identified differential expression of multiple genes associated with HLA class-I APM downregulation.<h4>Conclusions</h4>Our results reveal that APM downregulation is a prominent feature of NSCLC brain metastases, is independent from local IFNγ signaling defects and is associated with unfavorable clinical features. We also identified candidate modulators of the APM pathway in brain metastases with potential translational significance.

<h4>Background</h4>The Apolipoprotein-E ε4 (APOE4) allele is the strongest genetic risk factor for late-onset Alzheimer's disease (LOAD) and may contribute to neurodegeneration through a multi-hit hypothesis, in which vascular dysfunction, glial activation, and impaired lipid metabolism play central roles. Alterations in neurovascular unit (NVU) have emerged as an early APOE4-related phenotype, independent of amyloid and tau pathology. Astrocytes, as the primary source of APOE in the brain and key regulators of NVU homeostasis, may play a central role in these processes. This study investigates APOE4-associated NVU water exchange dynamics and astrocyte-vascular interactions using integrated in vivo MRI, ex vivo histology, and transcriptomic profiling.<h4>Methods</h4>Non-contrast multimodal MRI, including multi-echo time arterial spin labeling (multi-TE ASL), T1-weighted imaging, and diffusion-weighted MRI, were applied in 6-9-month-old APOE3-KI and APOE4-KI mice. Multi-TE ASL was used to estimate regional NVU water exchange dynamics, while diffusion MRI assessed tissue microstructural alterations. Immunohistochemistry evaluated perivascular matrix metalloproteinase-9 (MMP9) activity, vascular-associated markers, astrocytic AQP4 expression, and glial reactivity. Single-nucleus RNA sequencing (snRNAseq) characterized cell-type-specific transcriptional profiles, and inferred cell-cell communication analysis between astrocytes, pericytes, and other NVU components. Integrated analyses compared MRI-derived measures with molecular and cellular findings.<h4>Results</h4>APOE4-KI mice showed regionally specific alterations in NVU water exchange dynamics, particularly in the hippocampus, accompanied by trends toward altered microstructural complexity. Immunohistochemistry demonstrated increased perivascular MMP9 expression and evidence of extracellular matrix remodeling without prominent structural disruption of blood-brain barrier (BBB) markers in APOE4 mice. Astrocytes showed increased AQP4 expression, heightened proinflammatory gene signatures, and morphological reactivity. Molecular findings aligned with MRI, supporting the sensitivity of non-contrast MRI to early NVU alterations. Exploratory snRNAseq suggested an APOE4-enriched astrocyte subpopulation associated with immune activation and matrix-related pathways and suggested potential glial-vascular interactions that require validation in larger samples.<h4>Conclusions</h4>This integrated imaging and molecular analysis suggests that non-contrast multimodal MRI detects early APOE4-related changes in NVU exchange dynamics and glial-vascular interactions. By providing converging multiscale neuroimaging and cellular observations, this work provides a foundation for developing non-invasive biomarkers to monitor neurovascular vulnerability and guide early intervention strategies in individuals at risk for LOAD.

<h4>Background</h4>Huntington's disease (HD) involves progressive corticostriatal dysfunction, yet the temporal dynamics and cell type-specific vulnerability patterns remain incompletely understood. While recent single-cell studies in rapidly progressing models have revealed early developmental and regional changes, temporal profiling distinguishing pathogenic mechanisms from normal aging in full-length HTT models remains lacking. Resolving stage-specific temporal dynamics across interconnected striatal and cortical neuronal populations over protracted time is essential for identifying drivers of cellular dysfunction.<h4>Methods</h4>A temporal single-nucleus transcriptomic atlas was generated from striatum and motor cortex from heterozygous zQ175 knock-in mice at early symptomatic (6 months) and late symptomatic (18 months) stages. This full-length huntingtin model enables staging of progressive circuit dysfunction alongside physiological aging. The high inherited CAG repeat length of the zQ175 model places cells beyond the somatic expansion threshold associated with transcriptional dysregulation and identity erosion in vulnerable human neuronal populations, yet prior to the de-repression crisis and cell loss observed at the most extreme expansions in HD, providing a tractable window into the progressive molecular pathogenic cascade. Genotype-dependent effects were modeled to distinguish cell type-specific signatures of disease mechanisms from age-related and compensatory changes. Integration of weighted gene co-expression and transcription factor regulatory networks with protein-protein interaction databases predicted candidate regulators of stage-specific programs. Findings were validated across human HD datasets and the rapidly progressive R6/2 mouse model.<h4>Results</h4>Temporal gene and network analysis revealed diverging, converging and biphasic patterns of transcriptional changes, distinguishing progressive disease and neuronal identity loss from aging. 21 cell type-specific gene co-expression modules were validated in human HD and R6/2 mice datasets, revealing stage-specific shifts in cellular stress, proteostasis, and synaptic programs. Disease modules enriched for CAG repeat length-dependent genes resolved their temporal progression. Shared vulnerability across cortical and striatal projection neurons implicated epigenetic regulator Zswim6 and splicing factors Rbfox1 and Celf2 in corticostriatal dysfunction. Integrative network analysis identified Foxo1, Neurod2, and Npas2 as stage-specific transcriptional regulators. Cross-species validation established conserved gene regulatory modules in human HD, establishing generalizable cell type-specific gene modules of translational relevance.<h4>Conclusions</h4>This temporally resolved atlas reveals stage-specific transcriptional dynamics of disease-relevant gene expression programs and physiological trajectories in vulnerable neuronal populations. distinguished from aging alone. This work establishes an important framework for understanding the temporal and regional coordination of pathogenic mechanisms, providing molecular insights into stage-specific therapeutic intervention.

Marrow senescence contributes to overall organismal aging and involves functional alterations in both the mesenchymal and hematopoietic compartments of bone marrow. Although matrix remodeling-associated 7 (<i>MXRA7</i>) has been demonstrated to modulate mesenchymal function and megakaryocyte differentiation in mice, this study aimed to investigate the potential role of <i>MXRA7</i> in marrow senescence. Single-cell RNA sequencing was performed on bone marrow cells from young and aged wild-type and <i>MXRA7</i>-knockout mice. Comparative analysis of 2-month-old and 2-year-old mice revealed that aging significantly altered the cellular proportions within the bone marrow niche, and <i>MXRA7</i> deficiency markedly increased Macro1 macrophages in aged mice, likely driven by the dysregulation of the <i>Ccl24-Ccr3</i> axis. <i>MXRA7</i> deficiency altered Mid1 expression and the macrophage migration inhibitory factor (<i>Cd74 + Cxcr4</i>), <i>Ccl6 + Ccr2</i>, and Von Willebrand factor signaling pairs (<i>Itga2b + Itgb3</i>), all of which are closely associated with cell status in the bone marrow microenvironment. In summary, these findings underscore <i>MXRA7</i>'s role in cellular profile shifts during bone marrow aging, offering novel insights into how <i>MXRA7</i> coordinates hematopoietic and immune homeostasis in the bone marrow.

Cartilage is characterized by a highly specialized extracellular matrix (ECM) secreted by chondrocytes and limited self-regenerative capacity. In vivo investigations of chondrogenesis are limited by difficult and traumatic access, especially in humans. While it is known for decades that disturbances of chondrocyte differentiation and changed cartilage ECM composition cause severe skeletal phenotypes in vertebrates, a detailed molecular understanding of chondrogenesis and cartilage ECM formation is still missing, especially in the context of human genetic skeletal diseases. ATDC5 cells, derived from AT805 mouse teratocarcinoma cells, have been used in the past to model chondrogenic differentiation, however, most studies have investigated few major cellular differentiation markers only so that the composition of the secreted ECM as well as effects on the ATDC5 transcriptome upon differentiation are still unclear. Here, we performed time-resolved transcriptomic and ECM proteomic analyses of differentiating ATDC5 cells. Both datasets confirmed the formation of a cartilage-like matrix with increasing expression of key chondrocyte genes over the course of differentiation. ECM proteomics further revealed a number of ECM components not previously reported in ATDC5 cells or the secreted ECM, encompassing collagens, proteoglycans, glycoproteins and other secreted factors. Overall, our findings provide a more detailed molecular characterization of ATDC5 chondrogenesis and highlight the potential of this model system for ECM-focused studies.

Peripheral nerve injury (PNI), especially those with long-distance transected defects, remains a major clinical challenge due to limited regenerative capacity associated with inadequate endogenous Schwann cell (SC) support. Here, we developed a biomimetic nerve graft with a defined spatial gradient of immobilized SCs to facilitate effective transplantation and axonal guidance, thus enhancing peripheral nerve regeneration. SCs were initially encapsulated within decellularized nerve matrix (DNM) microgels using a customized flow-focusing microfluidic device. The DNM microgels supported good viability, facilitated cellular proliferation, and preserved the repair phenotype of the encapsulated SCs, while maintaining their advantageous paracrine activity conducive to axon extension. Furthermore, these microgels were incorporated into the density-gradient pores of a pre-designed scaffold, resulting in a gradient SC-laden scaffold, designated as MGs@G-scaffold. Notably, the spatially graded SCs within the MGs@G-scaffold created a stable environmental gradient of neurotrophic factors, thereby providing sustained biochemical cues to guide axonal elongation. Finally, when transplanted into a 15-mm rat sciatic nerve defect model, the SC-laden scaffolds exhibited significantly improved axonal regeneration, remyelination, and functional recovery compared with the SC-free scaffolds. Additionally, MGs@G-scaffolds outperformed scaffolds with uniform SC distribution (MGs@H-scaffolds), achieving therapeutic outcomes comparable to autografts. Overall, this study demonstrates that transplanting spatially graded SCs embedded in a tissue-engineered bioactive scaffold offers sustained ongoing support for cells, gradient biochemical signals, and a pro-regenerative microenvironment for nerve regeneration and functional recovery, which holds great promise for long-distance PNI repair.

Calcium (Ca²⁺) signaling is a key regulator of brain function and development. Here, we comprehensively analyze the Ca²⁺ signaling transcriptome in the adult mouse brain and the developing human brain to reveal the basis of signaling specificity. We show that neurons organize into non-stochastic Ca²⁺ states that reflect cell-type identity and capture subtle functional differences. These states arise from lineage-specific developmental Ca²⁺ programs that are detectable already in progenitor stages, and may precede differentiation into mature neuronal cell types. During neocortical development, many Ca²⁺ signaling genes, such as <i>ADGRV1, NCALD,</i> and <i>CREB5</i>, peak at distinct developmental stages, are evolutionarily conserved, and reflect transcriptional heterogeneity within progenitors associated with cell-fate decisions. Together, our findings provide an in-depth understanding of how a tightly regulated Ca²⁺ signaling transcriptome encodes cell-state-specific signaling programs and demonstrate that Ca²⁺ signaling is precisely tailored to distinct cell states.

Short-read RNA sequencing (RNAseq) remains a cornerstone for transcriptome profiling, but is limited in reconstructing full-length transcripts and capturing transcript diversity. While long-read RNAseq spans entire transcripts and resolves complex structures, this technology is hindered by its high error rates. In parallel, noncoding RNA transcripts remain underrepresented in current references. Here, we present hybrid <i>de novo</i> RNA assembly (HyDRA), a pipeline that integrates the accuracy of short reads with the structural resolution of long reads to produce more complete <i>de novo</i> transcriptome assemblies. Benchmarking showed HyDRA to outperform existing methods by up to 40%. Using the HyDRA human ovarian metatranscriptome, we identified >50,000 high-confidence long noncoding RNAs, most of which have not been previously detected using traditional methods. Although long-read RNAseq is advancing, the vast availability of short reads ensures HyDRA's ongoing role in capturing high-confidence, cell-type-specific transcripts and advancing our understanding of transcriptomic complexity and the noncoding genome.

<h4>Background</h4>Suicide is the second leading cause of death among children and adolescents, with rates of pediatric suicidal behavior rising substantially over the past two decades. The neurobiology of suicide has been extensively studied in adults, yet pediatric-specific evidence remains limited and the extent to which adult findings can be extrapolated to youth is unclear. This review synthesizes current evidence on the neurobiological correlates of suicidal ideation, suicide attempt, and death by suicide in pediatric and adolescent populations across neurological, genetic, epigenetic, inflammatory, metabolic, and endocrine domains.<h4>Methods</h4>A literature search was conducted in PubMed, Embase, PsycINFO, and Google Scholar for peer-reviewed, English-language human studies. Priority was given to pediatric and adolescent samples, with adult data included where pediatric evidence was lacking. Studies were grouped by biological domain and by suicidal phenotype.<h4>Results</h4>Suicidal ideation, suicide attempt, and death by suicide showed partially distinct biological signatures rather than lying on a single continuum of severity. Different markers, most notably cortisol regulation and stress-related DNA methylation, differed in direction between pediatric and adult cohorts, indicating that adult biomarker data cannot be directly extrapolated to youth. Findings converged on a developmental cascade in which genetic liability and early-life adversity influence the hypothalamic-pituitary-adrenal axis, with downstream effects on epigenetic regulation, neuroinflammation, neurochemistry, and frontolimbic circuitry.<h4>Conclusions</h4>Pediatric suicidal behavior reflects developmentally distinct biological processes that cannot be inferred from adult findings. Advancing the field will require longitudinal, multimodal pediatric studies that disaggregate suicidal phenotypes, span the pubertal transition, and apply age-stratified reference ranges, supporting biologically informed stratification and mechanism-targeted intervention.

Adaptive resistance limits durable benefit from immune checkpoint blockade (ICB) in the majority of cancer patients, yet the transcriptomic dynamics of the broader checkpoint landscape during treatment remain poorly characterized across tumor types. Here we present CheckDyn, a multi-cohort computational framework that profiles paired pre- and post-treatment transcriptomes to quantify treatment-induced changes across 38 immune checkpoint and exhaustion-associated genes and to predict adaptive resistance. We integrated publicly available RNA-seq and scRNA-seq data from 64 paired tumor samples spanning melanoma, basal cell carcinoma, and non-small-cell lung cancer (GSE91061, GSE120575, GSE123813, GSE176021), applying pseudo-bulk aggregation, Z-score batch correction, and Stouffer meta-analysis for cross-cohort harmonization. Paired Wilcoxon signed-rank testing and linear mixed-effects meta-analysis identified LAG3 (log₂FC = 0.596, padj = 0.015), PDCD1 (log₂FC = 0.810, padj = 0.003), TOX2 (log₂FC = 0.605, padj = 0.003), CD274 (log₂FC = 0.402, padj = 0.015), and IDO1 (log₂FC = 0.381, padj = 0.026) as consistently upregulated post-treatment across cohorts. Co-expression network analysis revealed extensive rewiring, with ENTPD1 (ΔDegree = +0.297) emerging as the largest hub-degree shift, suggesting a shift toward metabolic immune suppression. Temporal trajectory modeling showed that all 38 checkpoint genes followed linear upregulation trajectories, with PDCD1 and LAG3 carrying the steepest slopes. An ensemble classifier combining logistic regression and random forest on pre-to-post expression deltas achieved an area under the receiver operating characteristic curve (AUC) of 0.812 (95% CI: 0.694-0.930; 5-fold cross-validated AUC = 0.806) for adaptive resistance prediction across n = 59 patients with available response annotations. These findings establish a consistent transcriptional signature of compensatory checkpoint upregulation during ICB therapy and provide a data-driven framework for early identification of adaptive resistance that warrants external prospective validation.

<h4>Background</h4> Ectothermic vertebrates exhibit substantial physiological plasticity in response to environmental temperature fluctuations. Among them, amphibians show particularly pronounced metabolic adjustments under cold conditions; however, the molecular mechanisms by which the liver adapts to low temperatures remain poorly understood. To address this gap, we investigated the hepatic transcriptional response to cold exposure in the African clawed frog ( Xenopus laevis ) using RNA sequencing (RNA-seq) and quantitative PCR. <h4>Results</h4> Exposure to 5°C for five days in X. laevis resulted in pronounced hyperglycemia and extensive transcriptional reprogramming in the liver. RNA-seq analysis indicated that, relative to frogs maintained at 24°C, 2,392 genes were upregulated, whereas 2,031 genes were downregulated. Notably, genes associated with gluconeogenesis, such as foxo1 , g6pc1 , and pck1 , exhibited significant upregulation, whereas genes related to glycolysis and glucose utilization, including gck , pfkm , and ldhb , were downregulated. Concurrently, cold exposure induced the expression of genes involved in fatty acid synthesis, desaturation, and cholesterol biosynthesis, such as srebf1 , acaca , fasn , scd , fads2 , srebf2 , and hmgcr . Conversely, genes related to fatty acid β-oxidation, including ppara , cpt1a , cpt1b , slc25a20 , acadl , hadha , and hadhb , were significantly suppressed. In alignment with this pattern, multiple components of the mitochondrial electron transport chain and ATP synthesis machinery were also downregulated. Additionally, gene groups involved in antioxidant defenses, such as gpx4 , gpx1 , prdx2 , prdx5 , prdx6 , and ferritin-related genes, were upregulated. Representative transcriptomic changes were validated using quantitative PCR analysis. <h4>Conclusions</h4> Exposure to cold temperatures induced coordinated metabolic reprogramming in the liver of X. laevis . This reprogramming was characterized by the activation of gluconeogenesis, suppression of glycolysis, fatty acid β-oxidation, and oxidative phosphorylation, and induction of lipid and cholesterol biosynthetic pathways. These metabolic adjustments suggest that frogs acclimated to cold conditions adopt an energy-conserving metabolic strategy while sustaining glucose production and promoting lipid remodeling to adapt to low-temperature environments. These findings offer novel insights into the molecular mechanisms underlying cold adaptation in ectothermic vertebrates and underscore the pivotal role of the liver in managing transcriptional responses to cold stress.

<h4>Background</h4>Cisplatin is a first-line chemotherapeutic agent widely used in clinical oncology, but its clinical utility is severely limited by cisplatin-induced acute kidney injury (Cis-AKI). Renal tubular epithelial cells (RTECs) are the main target of cisplatin-induced damage, and the pathogenesis involves oxidative stress, inflammatory response, and multiple types of programmed cell death. As a core mitochondrial quality control mechanism, mitophagy is closely related to the above pathological processes, but its overall regulatory network in Cis-AKI remains to be systematically clarified.<h4>Main body</h4>This review systematically summarizes the core pathological mechanisms of cisplatin-induced nephrotoxicity, including mitochondrial dysfunction, oxidative stress, inflammation, apoptosis, pyroptosis, and ferroptosis. It focuses on the molecular pathways of mitophagy (PINK1/PARKIN-dependent and -independent pathways), the bidirectional crosstalk between mitophagy and the aforementioned pathological events, and the dual role of mitophagy in Cis-AKI. In addition, this review collates preclinical progress of mitophagy regulators and their renoprotective effects, and analyzes the current obstacles to clinical translation.<h4>Conclusion</h4>Mitophagy serves as a key regulatory node in Cis-AKI and can simultaneously ameliorate multiple injury pathways by clearing damaged mitochondria and reducing mtROS. Moderate mitophagy plays a renoprotective role, while excessive mitophagy aggravates renal injury. Targeted and precise regulation of mitophagy is expected to become a new strategy for the prevention and treatment of cisplatin-induced nephrotoxicity.

G protein-coupled receptor 52 (GPR52) has been identified as a promising therapeutic target for the treatment of schizophrenia, psychiatric disorders, and other human neurological diseases. A series of novel 2,3-disubstituted pyrazine derivative GPR52 agonists have been designed, synthesized, and biologically evaluated. Through the exploration of the structure-activity relationship, several potent GPR52 agonists (<b>14a</b>, <b>14h</b>, <b>14l</b>, <b>20c,</b> and <b>20i</b>) were identified, with EC₅₀ values in the nanomolar range (26-70 nM). Further studies on <b>14h</b> indicate that it has excellent pharmacokinetic properties in mice and rats, as well as brain permeability (brain/plasma ratio = 1.6 and 3.1, respectively). <i>In vivo</i>, compound <b>14h</b> (<b>R-185</b>) significantly inhibits MK-801-induced hyperlocomotor behaviors in zebrafish larvae and mice. Overall, our research results have identified an effective, orally bioavailable, and brain-permeable GPR52 agonist (<b>14h</b>), which is a promising candidate for the development of antischizophrenia drugs.

<h4>Background</h4>Iron overload remains a critical complication secondary to transfusional therapy and altered iron metabolism in patients with sickle cell anemia (SCA). Hyperferritinemia often serves as a surrogate marker for iron overload; however, as an acute-phase reactant, it may be elevated due to chronic inflammation, infection, or liver disease, potentially overestimating true iron stores. In Yemen, the burden of SCA is high, yet local data on the frequency of hyperferritinemia are scarce. This study aimed to describe the distribution of serum ferritin (SF) levels and the proportion of SCA patients with hyperferritinemia among adolescents and young adults attending the YSTH clinic in Sana'a, and to explore associations with hematological, and sociodemographic parameters.<h4>Methods</h4>A prospective cross-sectional study was conducted between June and December 2024, involving 80 participants (aged ≥ 15 years). The data were analyzed using SPSS version 26. Due to the non-normal distribution and high dispersion of SF, the Median (IQR) and Spearman correlation were utilized. Bivariate analysis calculated crude odds ratio at (95% CI) to identify predictor of elevated SF. Statistical power was limited by the sample size (N = 80) to detect weak associations.<h4>Results</h4>The study enrolled 80 SCA patients with a mean (±SD) age of 18.5 (±3.69) years, of whom 65% were males and 35% were females. The median (IQR) SF level was 1645 ng/mL (414-3525), with values ranging from 77.1 to 4500.0 ng/mL. Approximately 85% of patients exhibited elevated SF level (≥300 ng/mL). While red cell indices did not significantly predict the risk of hyperferritinemia, a significant positive correlation was observed between SF levels and white blood cell count (p = 0.03), suggesting a possible link with the inflammatory state. No significant associations were found between sociodemographic factors and high SF levels.<h4>Conclusion</h4>A high prevalence (85%) of hyperferritinemia (≥ 300 ng/mL) was observed among SCA patients attending the YSTH in Sana'a, Yemen, suggesting probable iron overload. While findings support routine SF screening but highlight the need for future studies incorporating inflammatory markers (e.g., CRP) and systematic transfusion histories to definitively quantify the iron overload burden in this population.

<h4>Background</h4>Accurate identification of gene fusions is critical for precision oncological approaches to sarcomas, where specific fusion genes are actionable targets of tyrosine kinase (TK) inhibitors. This study provides a descriptive overview of the real-world use, under a universal health insurance program, of 3 different comprehensive genomic profiling (CGP) panels within the Japanese national CGP system: GenMineTOP, which integrates DNA and RNA sequencing, and 2 DNA-based panels, FoundationOne CDx and the OncoGuide NCC Oncopanel System.<h4>Methods</h4>We examined 2,633 sarcoma cases from the Center for Cancer Genomics and Advanced Therapeutics (C-CAT) database to identify potentially actionable alterations classified by the Cancer Knowledge Database (CKDB). The mean patient age was 51.1 years, and 45% were female. Detection rates of TK fusions (FGFR1-4, NTRK1-3, ALK, RET, ROS1, and BRAF) were compared descriptively. Additionally, 3 sarcoma cases with an NTRK fusion detected by GenMineTOP were functionally validated.<h4>Results</h4>Potentially actionable alterations (CKDB Levels A, B, and C) were identified in 566 patients (21.5%). Actionable TK fusions were detected in 3.1% (8 of 254) with GenMineTOP, 1.4% (31 of 2,211) with FoundationOne CDx, and 0.6% (1 of 168) with NCC Oncopanel. GenMineTOP identified rare NTRK fusions, including RBPMS::NTRK2, HTT::NTRK2, and EML4::NTRK3. Functional assays suggested their oncogenic potential and demonstrated responsiveness to the TK inhibitors larotrectinib and entrectinib.<h4>Conclusions</h4>In this cohort, the DNA+RNA-based panel showed a higher detection rate for TK fusions, although the panels were applied to different patient groups and the sensitivity and specificity could not be determined. Future studies evaluating different test types on the same tumor specimens are warranted to clarify whether the dual-sequencing approaches can improve the identification of actionable genetic events.<h4>Level of evidence</h4>Diagnostic Level IV. See Instructions for Authors for a complete description of levels of evidence.

Severe traumatic brain injury (sTBI) is a leading cause of edema and neurological dysfunction. However, the molecular mechanisms driving injury progression remain poorly understood. Here, we present a single-nucleus transcriptomic atlas of human and male mouse cortex across multiple sTBI stages, providing a systematic landscape of dynamic, cell-type-specific responses to injury and edema. Cross-species analyses reveal both conserved and human-specific programs, including a polarized oligodendrocyte state absent in murine sTBI. This state shows increased ABCA1 and adhesion-related genes and is associated with edema progression. In vitro experiments support an ABCA1-linked role in lipid handling that may support oligodendrocyte-lineage cell function, reflected by the maintenance of myelin-associated marker expression. Analyses combining ligand-receptor interactions, co-expression, and perturbation experiments link microglial CALM2-HMGB1 signaling to oligodendrocyte ABCA1-associated lipid regulation during injury progression. These human-specific programs indicate that rodent models may not fully recapitulate human sTBI mechanisms, underscoring the need for caution when extrapolating cross species findings.

Cardiac fibrosis in congenital heart disease (CHD) is associated with poor outcomes, but the genetic risk factors have not been clearly outlined. This review details genes important for regulation of normal cardiac development or fibrosis, particularly cilia-related genes. Specific CHD pathologies have different patterns of fibrosis, likely from interaction between genetic mutations and environmental factors. Future studies are more feasible as tools like single-cell RNAseq and patient-derived organoids have become more affordable and easier to implement. A better understanding of genetic risk factors for fibrosis in CHD could improve diagnosis and treatment for these patients.

Mining activities are a major source of heavy metal contamination in agricultural systems, posing risks to food safety and human health. This study investigated the soil-rice system in a zinc (Zn) mining-affected area of western Thailand to: (1) quantify heavy metal contamination across soil and rice compartments; (2) evaluate compartment-specific transfer factors (soil-root, root-stem, and stem-grain) to elucidate metal mobility and accumulation pathways; and (3) identify soil properties influencing metal retention and mobility. Soil, root, stem, and grain samples were collected from 20 paddy fields and analyzed for As, Cd, Cr, Cu, Mn, Pb, Zn, Hg, and Fe. The results showed that although bulk soil concentrations were within national standards, the geo-accumulation index (I_geo) indicated extremely severe Cd and Hg contamination at all sites, with Zn also reaching extremely severe levels at several locations. In addition, consistently elevated pollution load index (PLI) values (> 1) indicated widespread multi-metal pollution across the entire study area. Soil organic matter, cation exchange capacity, and texture were strongly correlated with heavy metal retention and transfer (p < 0.05). Transfer factor analysis revealed high root accumulation of As and Hg, restricted Pb and Cd mobility, and substantial grain translocation of Cu, Zn, and Mn. Despite limited mobility, grain concentrations of As (1.16 mg kg^-1), Cd (0.98 mg kg^-1), Pb (4.61 mg kg^-1), and Hg (0.08 mg kg^-1) exceeded national food safety standards. Health risk assessment showed that both children and adults faced extremely high non-carcinogenic and carcinogenic risks from rice consumption, with hazard index and cancer risk values exceeding safe thresholds by several orders of magnitude. These findings demonstrate that the legacy of mining activities continues to threaten food safety and public health through rice-based exposure, highlighting the urgent need for targeted remediation, and routine monitoring to mitigate risks and support sustainable agriculture in mining-impacted regions.

Dysregulation of IDH1, a key metabolic enzyme, is widely reported across cancers and affects both tumor biology and patient prognosis. However, its regulation and associated epigenetic alterations in breast cancer (BC) remain unclear. This study examined methylation of two CpG islands (I1, I2) within the IDH1 promoter and a putative distal enhancer in clinical BC samples, correlating these patterns with IDH1 expression and clinicopathological features. Expression of PFKP, HIF-1α, and SIX1 was profiled to assess metabolic and developmental pathways. Additionally, genome-wide methylation analysis of IDH1-stratified TCGA-BRCA cohorts was performed to identify topological probe clusters with potential regulatory relevance. Twelve CpGs within I1 (- 109 to - 205 bp) were significantly hypermethylated in BC and showed an association with reduced IDH1 expression and a trend toward improved survival in this cohort, outperforming IDH1 expression in ROC analysis. I1 methylation was markedly higher in postmenopausal, metastatic, and TNBC cases and mapping to key transcription factor binding sites (CTCF, MYC, STAT3), suggesting disruption of regulatory complexes. PFKP, HIF-1α, and SIX1 were upregulated in these cases, indicating metabolic and developmental reprogramming. Genome-wide methylation analysis of IDH1-stratified TCGA cohorts revealed widespread alterations, with 33% of differentially methylated probes mapping to intergenic regions. High-density clusters on chromosomes 13 (85.7/kb) and 6 (32.5/kb), along with hypermethylation of CLDN18 and HRNBP3 promoters, implicated chromatin remodeling and splicing as frequent IDH1-associated events. Network analysis identified 332 hub genes, including HNRNPA1 and CBX5, reinforcing the role of epigenetic deregulation in BC pathogenesis. These findings suggest that IDH1 promoter methylation may hold prognostic relevance and suggest that altered IDH1 regulation contributes to broader epigenetic and chromatin remodeling events in BC.

Despite intensive glycemic control, diabetic cardiomyopathy (DCM) often progresses due to hyperglycemic memory (HGM), yet the specific cardiac cells perpetuating this injury remain unknown. To address this, we performed single-nucleus RNA sequencing (snRNA-seq) on hearts from an HGM rat model. Our analysis of 86,120 nuclei revealed HGM-specific inflammatory and epigenetic reprogramming signatures. Fibroblasts emerged as potential mediators, appearing to drive extracellular matrix remodeling via upregulated LAMININ and COLLAGEN signaling. We uncovered a distinct HGM-specific fibroblast subpopulation characterized by oxidative stress and H3K27 demethylation. Integrative analysis prioritized Fmo2 as a key pathogenic candidate, which was further supported via Mendelian randomization and clinical data as a putative causal gene. This study suggests that a pathogenic Fmo2⁺ fibroblast subpopulation may act as a pathological "memory carrier," providing novel mechanistic insights and proposing exploratory therapeutic avenues for HGM-induced cardiac damage beyond glycemic control.

<h4>Background</h4>Congenital aniridia is a rare disorder presenting as a panocular malformation with variable severity, often complicated by progressive keratopathy. The purpose of this study was to characterise the tear-film proteome in adults with PAX6-related congenital aniridia and to identify dysregulated pathways linked to aniridia associated keratopathy (AAK).<h4>Methods</h4>Tears were obtained with Schirmer strips from four genetically confirmed patients and four age- and sex-matched healthy volunteers. Peptides prepared with the single-pot, solid-phase-enhanced (SP3) protocol were analysed by data-independent nanoLC-MS/MS. Proteins were identified with a false discovery rate (FDR) <1% during DIA data processing. Differential abundance between controls and patients samples was assessed using an adjusted p-value < 0.05. Proteins with |log₂-fold change| ≥1 were considered significantly expressed. Functional enrichment was evaluated with Enrichr (Gene Ontology, Reactome, JensenExp, Orphanet, TissueExp).<h4>Results</h4>A total of 3 162 proteins were detected; 2 633 showed a valid intensity in every sample of at least one group and were retained for statistical testing. Seventy-three (2.8%) were differentially expressed: 33 were over-expressed and 40 under-expressed in aniridia tears. Down-regulated proteins clustered in lipid homeostasis, epithelial junction integrity and wound-healing modules and included lacritin, secretoglobins and cytoskeletal adaptors, indicating a fragile, poorly repaired surface. Up-regulated species were dominated by neutrophil effectors (CD177 ≈ 50-fold) and reflected heightened innate immunity and abnormal epithelial maturation. Anti-angiogenic processes were significantly over-represented in both under and over-expressed protein sets.<h4>Conclusion</h4>Our workflow proved highly sensitive, capturing more than 3 000 tear proteins and thus underscoring the robustness of our proteomic approach. The tear film in aniridia reflects dysregulation of various processes, including immunity, lipid and epithelial homeostasis, and vascular remodelling. Our approach highlights novel biomarkers critical for developing targeted therapeutic strategies.<h4>Trial registration</h4>ClinicalTrials.gov, NCT05562115. Registered on 29 September 2022.

<h4>Background</h4>Mineral trioxide aggregate (MTA) is a calcium silicate-based endodontic biomaterial with well-established sealing ability, biocompatibility, and hard-tissue induction potential; however, the extent to which its biological performance can be modulated by nanostructured reinforcements remains insufficiently defined. Hexagonal boron nitride (hBN) is a layered two-dimensional nanomaterial with high chemical stability, mechanical robustness, and growing relevance in regenerative biomaterials. Because no directly comparable in vivo study had evaluated hBN incorporation into an MTA matrix in a standardized rat tibial defect model, the present work was designed as an exploratory physicochemical and histological study.<h4>Methods</h4>Standardized cylindrical defects (2.5 mm diameter, 4 mm depth) were created in the right tibial metaphysis of 40 systemically healthy rats randomly allocated to four groups (n = 10/group): empty defect control, pure MTA, MTA + 5 wt% hBN, and MTA + 10 wt% hBN. The study was conducted as a single 8-week in vivo experiment following institutional ethics approval. Pure MTA and hBN-modified composites were prepared under standardized mixing conditions and characterized qualitatively by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM), and energy-dispersive X-ray spectroscopy (EDX). After 8 weeks, tibiae were harvested, decalcified, paraffin-embedded, stained with hematoxylin-eosin, and scored semi-quantitatively for fibrous tissue, new bone formation, and osteoblastic cell presence. Non-parametric comparisons were performed using Kruskal-Wallis and Dunn-Bonferroni tests (α = 0.05).<h4>Results</h4>XRD and FT-IR analyses showed preservation of the principal calcium silicate phase architecture and hydration-related bonding environment of MTA after hBN incorporation, while SEM-EDX demonstrated a progressively denser and more hBN-rich surface organization in the modified groups, particularly at 10 wt% hBN. Histologically, both hBN-containing groups displayed significantly higher new bone formation and osteoblastic cell presence scores than the empty-defect control. Fibrous tissue scores were also higher in the hBN groups than in the control group, indicating that the tissue response should be interpreted as a modified fibro-osseous reparative profile rather than as simple suppression of fibrosis. Compared with pure MTA, the 10 wt% hBN formulation showed the most favorable overall trend within the tested range, although differences between 5 wt% and 10 wt% hBN were not statistically significant.<h4>Conclusions</h4>Within the boundaries of this exploratory 8-week rat tibial defect study, hBN could be incorporated into MTA without evidence of major physicochemical destabilization and with a more pronounced bone-related healing response than that observed in untreated control defects. The findings do not support an overextended claim of direct osteoblast activation or definitive concentration optimization; rather, they indicate that hBN-reinforced MTA is a promising proof-of-concept composite whose long-term remodeling behavior, mechanical performance, ion release, systemic safety, and true optimum hBN window require further quantitative investigation.

Gallbladder cancer (GBC) is a highly aggressive malignancy with late presentation, limited therapeutic options, and dismal survival rates. Despite recent advances in molecular profiling, our understanding of GBC remains fragmented, with most studies focusing on isolated genetic or epigenetic events rather than integrating the broader molecular interplay. Frequently altered genes-including TP53, KRAS, ERBB2, and CDKN2A-impact critical signaling cascades such as Ras/MAPK, PI3K/AKT/mTOR, Wnt/β-catenin, and p53, with aberrant ERK pathway activation emerging as a pivotal driver. Epigenetic modifications, dysregulated non-coding RNAs, and immune microenvironmental factors add further layers of complexity but remain underexplored in a unified context. Currently, no comprehensive framework links these alterations into a generalized molecular mechanism that could accelerate biomarker discovery and therapeutic innovation. This review integrates genomic, epigenomic, and immunologic data, complemented by KEGG pathway enrichment analysis, to present a consolidated molecular landscape of GBC. By mapping converging oncogenic hubs and signaling cross-talk, we aim to identify actionable targets-particularly within the ERK axis-that could inform early detection, guide precision therapy, and direct future translational research.

<h4>Objective</h4>This article provides a systematic review of the biological characteristics, regulatory mechanisms, and roles of MicroRNA-7 (miR-7) in the nervous system, as well as explores its clinical translation potential and challenges.<h4>Methods</h4>A review analysis integrating the progress of basic and clinical research related to miR-7.<h4>Results</h4>Characteristics and Regulation: miR-7 is highly expressed in mammalian brain tissues, with its function finely regulated by ceRNA networks (such as ciRS-7 and lncRNA SNHG1) and epigenetic modifications, forming a multi-layer dynamic regulatory system.<h4>Mechanisms of action</h4>It plays multi-faceted intervention roles in protein aggregation, neuroinflammation, mitochondrial dysfunction, and tumor progression by targeting key factors such as α-synuclein, NLRP3 inflammasome, EGFR/PI3K/AKT/mTOR pathway, and mitochondrial-related proteins (e.g. VDAC1). Role in Disease: It has significant pathophysiological implications in diseases such as Parkinson's disease, Alzheimer's disease, ischemic stroke, cerebral hemorrhage, and glioblastoma. Translational Potential: Changes in miR-7 expression in bodily fluids (blood, cerebrospinal fluid) and extracellular vesicles demonstrate diagnostic and prognostic potential; delivery systems based on nanomedicine (e.g. liposomes, graphene oxide, AAV vectors) and their combination therapy strategies (e.g. in conjunction with chemotherapy and immunotherapy) enhance its brain targeting and therapeutic efficiency.<h4>Conclusion</h4>miR-7 is a key regulatory molecule in brain diseases, with significant value for basic research and clinical translation. Future advancements should leverage cutting-edge technologies such as single-cell sequencing, spatial transcriptomics, and intelligent responsive nanocarriers to deepen the understanding of its regulatory networks and address challenges such as mechanism complexity, delivery system targeting, and the lack of clinical translation standards, to accelerate its transition from basic research to clinical application.

Global disability from osteoarthritis (OA) remains a staggering burden, yet disease-modifying pharmacological therapies remain limited. Current evidence supports OA as a whole-joint disease in which subchondral bone remodeling acts as an early and active driver of structural progression and pain. Osteoclasts are key regulators of this process, but clinical trials targeting osteoclast-dependent remodeling have yielded inconsistent results. This translational gap suggests that the traditional view of osteoclasts as a homogeneous, terminally differentiated resorptive population is no longer sufficient. Latest breakthroughs in single-cell and spatial multi-omics have begun to redefine osteoclast biology by revealing heterogeneity across precursor origin, lineage state, functional output, and niche-specific adaptation. In OA, these studies have clarified the osteoclastogenic microenvironment more clearly than the terminal taxonomy of mature osteoclast subsets, thereby shifting the field toward a state-spectrum framework. In this review, we synthesize recent high-resolution evidence to examine how osteoclast-lineage heterogeneity is organized across disease stages and osteochondral microenvironments, and how distinct osteoclast-lineage programs contribute to subchondral remodeling, angiogenic coupling, interface instability, and pain-related pathology. We further discuss how this framework may inform patient stratification, mechanism-matched intervention, and the development of program-specific therapies in OA.

<h4>Background/objectives</h4>Epithelial ovarian cancer (EOC) is the deadliest gynaecologic malignancy, largely due to late-stage diagnosis and ineffective therapy. EOC commonly spreads through the peritoneal cavity as multicellular spheroids, which are metastatic structures that enhance survival under detachment stress, promote dissemination, and contribute to therapeutic resistance. We previously showed that ULK1, a serine/threonine kinase classically linked to macroautophagy initiation, supports EOC progression, suggesting non-canonical roles in spheroid biology and pathogenesis.<h4>Methods</h4>CRISPR/Cas9 <i>ULK1</i> knockout (<i>ULK1</i>KO) models were generated in OVCAR8, HEYA8, and ES2 cells. Mitochondrial degradation phenotypes were assessed in spheroids by immunoblotting and fluorescence microscopy. Label-free proteomics with bioinformatic pathway analysis identified ULK1-associated programs in EOC spheroids. Bioenergetic consequences were quantified using Seahorse ATP-Rate assays. Therapeutic interactions were evaluated using multi-dose combination matrices testing the ULK1 inhibitor DCC-3116 with metformin.<h4>Results</h4>ULK1 modulated mitochondrial degradation in a cell-line-specific manner, either promoting or protecting against mitochondrial loss through mechanisms that were uncoupled from canonical autophagy machinery. Proteomic and bioinformatic analyses revealed significant alterations in mitochondria-related processes, aligning with emerging ULK1 functions in mitochondrial homeostasis. <i>ULK1</i> loss broadly reduced OXPHOS complex proteins in EOC spheroids and consistently decreased hexokinase 2 (HK2), indicating coordinated metabolic remodeling. Seahorse profiling mirrored these shifts: OVCAR8 <i>ULK1</i>KO spheroids showed reduced OCR and ATP production, whereas HEYA8 and ES2 <i>ULK1</i>KO spheroids exhibited increased mitochondrial ATP production. Combination matrices showed potential synergy between DCC-3116 and metformin.<h4>Conclusions</h4>These data show that ULK1 differentially regulates mitochondrial degradation across EOC spheroid models through potential mechanisms alternative to canonical autophagy machinery, while reshaping spheroid metabolism and revealing potential therapeutic vulnerabilities in advanced EOC.

<b>Background</b>: The insulin-like growth factor 1 receptor (IGF1R) is a receptor tyrosine kinase whose both overexpression and underexpression have been implicated in the initiation and progression of breast tumorigenesis. The mechanism through which underexpression of the receptor contributes to a more aggressive phenotype is currently less understood. <b>Methods</b>: Through the expression of a dominant-negative IGF1R, we studied the phenotypic effects of receptor inhibition on early MMTV-<i>Wnt1</i> mouse mammary tumors. Utilizing histopathological techniques and single-cell RNA-sequencing, we explored cellular heterogeneity and transcriptional alterations that occur as a result of IGF1R inhibition. <b>Results</b>: Examination of primary tumors failed to reveal obvious differences in tissue architecture or expression of differentiation markers with IGF1R inhibition. Both cohorts of tumors produced metastatic lesions in the lung. Single-cell RNA-sequencing identified previously unknown epithelial subpopulations that were present in both tumor types. In tumors with inhibited IGF1R, a previously undescribed epithelial population marked by expression of both <i>Krt14</i> and <i>Krt6a</i> was identified, transcriptionally distinct from its MMTV-<i>Wnt1</i> counterpart, and present in the smallest lung metastases. In human breast cancer patients, expression levels of <i>KRT14</i> and <i>KRT6A</i> negatively correlated with expression of <i>IGF1R</i>. <b>Conclusions</b>: Inhibition of the IGF1R in a mouse model of basal-like breast cancer produces transcriptionally distinct <i>Krt6a⁺</i>/<i>Krt14⁺</i> epithelial cells, which are present in the smallest metastatic lesions identified in the lung. Expression of genes associated with this population may potentially be effective biomarkers of metastatic capacity in basal-like breast tumors with low levels of IGF1R expression.

Fatty acids (FAs) have drawn attention in the field of oncology due to their multifaceted role, not only as structural components of lipid-based delivery systems but also as functional moieties that can enhance the pharmacokinetic and biological behavior of anticancer drugs and, subsequently, their therapeutic performance. Due to their biocompatibility, structural diversity, high affinity for biological membranes, and albumin-binding capacity, FAs can increase drug lipophilicity, membrane permeability, systemic distribution, tissue distribution, and enable controlled enzymatic release. All these properties endorse the development of nanocarriers containing FAs, such as liposomes, lipid nanoparticles (LNPs), self-nanoemulsifying drug delivery systems (SNEDDS), and self-assembling lipidic prodrugs (LAPs). In addition, several FAs, especially polyunsaturated FAs, seem to have a direct anticancer activity by modulating lipid metabolism, oxidative stress, membrane organization, and regulating cell death pathways. This review summarizes the FA conjugation chemistry, the influence of FA on the pharmacokinetics and tumor-targeting capacity of anticancer agents, and the current developments in FA-based cancer treatment strategies, while also covering the biological functions of FA in cell death pathways and cancer metabolism. By integrating medicinal chemistry, nanocarrier design, pharmacokinetic modulation, and tumor lipid biology, this review positions FA-based strategies as a relevant and evolving platform for improving anticancer drug delivery, tumor selectivity, and therapeutic performance.

<h4>Background</h4>Cutaneous squamous cell carcinoma (cSCC) is one of the most common skin malignancies with limited reliable biomarkers for early detection and prognosis. This study integrated bulk and single‑cell transcriptomic data to identify post‑translational modification-related key genes and to characterise cellular and immune alterations in cSCC.<h4>Methods</h4>Publicly available cSCC datasets and post-translational modification-related genes (PTMRGs) were analysed. Single-cell analysis identified key cell populations and differentially expressed genes (DEGs), which were intersected with bulk DEGs, weighted gene co‑expression network modules, and PTMRGs. Candidate genes were prioritised using five feature selection algorithms, validated by quantitative real-time polymerase chain reaction (qRT-PCR) and receiver operating characteristic (ROC) analysis [area under the curve (AUC) >0.7]. Functional exploration included predictive modelling, SHapley Additive Explanations (SHAP) interpretation, immune infiltration analysis, protein interaction mapping, drug prediction with molecular docking, intercellular communication inference, and pseudotime trajectory analysis.<h4>Results</h4>Keratinocytes were identified as the key cell population, yielding 999 single-cell-differentially expressed genes (sc-DEGs). Bulk analysis detected 5,771 DEGs and 17 co‑expression modules; intersection yielded 19 candidate genes. Multi‑method feature selection nominated HIF1A, UBC, and GSTP1; validation retained HIF1A and GSTP1 as final diagnostic markers (upregulated in cSCC; AUCs >0.7). A least absolute shrinkage and selection operator (LASSO) classifier achieved the best performance. SHAP highlighted HIF1A and GSTP1 as dominant contributors. HIF1A was predicted to be nuclear, GSTP1 mitochondrial; both correlated positively with macrophages, natural killer (NK) cells, and neutrophils. Drug prediction and docking identified camptothecin, hydroquinone, and resveratrol with favorable binding (e.g., HIF1A-camptothecin -8.7 kcal/mol; GSTP1-camptothecin -8.6 kcal/mol). Cell-cell signaling was enhanced in cSCC [vascular endothelial growth factor (VEGF), endothelin (EDN) pathways; elevated MIF-(CD74+CXCR4) and MIF-(CD74+CD44)]. Keratinocyte pseudotime revealed progression from normal to metastatic states with concordant keratinocyte‑related gene programs.<h4>Conclusions</h4>HIF1A and GSTP1 are promising diagnostic biomarkers for cSCC, linked to altered immune landscapes, enhanced intercellular signaling, and potential druggable interactions.

<h4>Background</h4>Hepatocellular carcinoma (HCC) continues to pose a major health concern for global public health, characterized by pronounced clinical heterogeneity in survival outcomes and therapeutic efficacies. The stromal senescence constitutes a major impediment in constructing robust prognostic models. This study aimed to develop and validate a robust prognostic prediction model to accurately assess the prognostic risk of HCC patients and facilitate personalized precision treatment.<h4>Methods</h4>We obtained data from the Gene Expression Omnibus (GEO), The Cancer Genome Atlas (TCGA), and the Clinical Proteomic Tumor Analysis Consortium (CPTAC) databases. Area Under the Curve Cell (AUCell) was used to quantify senescence in single-cell RNA sequencing (scRNA-seq) of HCC. A stromal-senescence prognostic signature was then built by using Cox and least absolute shrinkage and selection operator (LASSO) analyses. High- and low-risk stratification was interrogated with clusterProfiler, maftools, immune-checkpoint analysis, and oncoPredict. Signature expression was cross-validated in scRNA-seq, bulk RNA sequencing (RNA-seq), proteomic, and spatial transcriptomic datasets. Small-molecule ligands targeting DNASE1L3 were identified by virtual screening, and their binding was verified by molecular docking and dynamics simulations.<h4>Results</h4>It was found that HCC-associated stromal cells exhibited the most pronounced levels of senescence among cell populations. Using machine-learning algorithms, we successfully established <i>DNASE1L3</i>, <i>NDRG2</i>, <i>ADAM15</i>, <i>IGFBP3</i>, <i>MMP14</i>, and <i>ANGPT2</i> as a stromal-senescence prognostic signature in HCC. Subsequent comparisons between patient subgroups based on this signature revealed distinct differences in long-term survival outcomes, mutational landscapes, immune checkpoint expression, and responsiveness to chemotherapeutic agents. Further, the stromal-senescence prognostic signature is significantly associated with metabolic reprogramming, immune inflammation, cell proliferation, and metastasis. Multi-omics data analyses were employed to validate the expression levels of this signature in both tumor and normal tissues. In addition, virtual screening identified three potential <i>DNASE1L3</i> candidates: STL167676, STK598132, and STK119225.<h4>Conclusions</h4>This study developed a robust stromal senescence-associated gene signature that accurately predicts survival outcomes in HCC patients and identified candidate stromal senescence-related molecules for more effective personalized therapy.

Lung cancer is a leading cause of global cancer mortality, with <i>EGFR</i> mutations serving as a primary therapeutic target. Although <i>EGFR</i> tyrosine kinase inhibitors (TKIs) are initially effective, acquired resistance inevitably develops. While <i>ROS1</i> rearrangements are well-known baseline drivers, they are exceptionally rare as acquired resistance mechanisms. We utilized next-generation sequencing (NGS) to identify a rare ROS1 intragenic rearrangement (exons 35-37) in three never-smoking women with <i>EGFR</i>-mutant lung adenocarcinoma following progression on <i>EGFR</i> TKIs. Clinical courses were heterogeneous: one patient achieved a durable partial response using combined osimertinib and crizotinib. A second patient, intolerant to dual TKI therapy due to QTc prolongation and grade 3 edemas, achieved a sustained partial response with platinum-pemetrexed chemotherapy. The third patient exhibited polyclonal resistance, including <i>EGFR</i> C797S and <i>TP53</i> mutations, with fatal central nervous system progression. In this three-patient case series, <i>ROS1</i> exon 35-37 RNA-level intragenic rearrangements were repeatedly detected after <i>EGFR</i>-TKI progression, suggesting a rare transcript-level alteration within heterogeneous resistance evolution. However, its biological significance, driver versus passenger role, and therapeutic relevance remain uncertain. Combined <i>EGFR</i> and <i>ROS1</i> inhibition may be considered in selected cases, but further validation is required.

<h4>Background and aims</h4>The American Gastroenterological Association (AGA) Clinical Care Pathway provides a tiered, noninvasive algorithm for fibrosis risk stratification in populations at high risk for metabolic dysfunction-associated steatotic liver disease (MASLD). We assessed the 2-year longitudinal performance of the AGA pathway using magnetic resonance elastography (MRE) as the reference.<h4>Methods</h4>This prospective cohort study enrolled adults aged 50-79 years with type 2 diabetes mellitus from ambulatory care clinics between the dates of February 2016 and February 2025. Participants underwent a standardized clinical research visit with Fibrosis-4 index (FIB-4), vibration-controlled transient elastography (VCTE), and MRE at baseline and at a 2-year interval at the UCSD MASLD Research Center.<h4>Results</h4>Of 626 participants with a baseline assessment, 209 had longitudinal follow-up assessment and were included in the study. The prevalence of MASLD was 69.9%, and 19.0% had significant fibrosis at baseline (MRE ≥3.30 kPa). Applying the AGA pathway of FIB-4 and VCTE, the false negative rate (low risk by pathway with MRE ≥3.30 kPa) at baseline was 7% with 17.6% of participants qualifying for specialty referral. At 2-year follow-up, the false negative rate decreased to 3% and an additional 7%, respectively qualified for specialty referral. Applying a FIB-4 cut point of 1.0 decreased the false negative rate to 0%; however, the number of patients requiring VCTE increased by 54% over 2-years.<h4>Conclusion</h4>Longitudinal reassessment of patients initially classified as low risk by the AGA Clinical Care Pathway substantially reduced misclassification of significant fibrosis, while maintaining a low rate of specialty referral. These findings support serial re-evaluation as a key component of noninvasive fibrosis risk stratification in at-risk populations.

<title>Abstract</title> <p>Respiratory virus infections affect both humans and livestock, causing considerable mortality and morbidity. While respiratory pathogens such as swine influenza A virus (pH1N1) and porcine respiratory coronavirus (PRCV) often present with overlapping clinical symptoms, their pathological trajectories and outcomes differ. Given the propensity for pathogen spillover and the use of pigs as a physiologically relevant large-animal translational model, we aimed to characterise host serum protein signatures that detect and differentiate pH1N1 from PRCV, enabling improved disease monitoring and control. Using high-resolution mass spectrometry-based proteomics, we identified 162 serum proteins that were significantly dysregulated across 3 infection timepoints (1, 5, and 12 days post-infection (DPI)), with signatures correlating with viral shedding and lung pathology as early as 1 DPI. Notably, multiplexed targeted analysis of a subset of proteins in an independent cohort from a different breed and geographic location demonstrated detection, femtomole-level targeted quantitation, and validation of SRGN as a diagnostic marker for pH1N1 and PRCV (AUC = 0.85). Further, SOD1 was validated as an early marker for PRCV, increasing as early as 1 DPI (AUC = 0.9). Finally, a multi-peptide signature composed of SRGN, SOD1, and RAN demonstrated reasonable predictive power for pH1N1 (AUC = 0.75) and PRCV (AUC = 0.65) at 1 DPI. Our data validate the proteomic screening, provide insights into the role of early protein markers in distinguishing respiratory viral infections, and pave the way for the development of point-of-care diagnostics and targeted prevention strategies, enhancing preparedness against emerging zoonotic threats.</p>

The choroid is critical for maintaining vision and implicated in several ocular diseases, being the sole source of nutrients and waste removal for the outer retina. Genetic discovery can help elucidate the pathways through which choroidal features influence disease risk. Our meta-analysis of genome-wide association studies (n= 78,682 participants) identified 30 genomic regions, including 20 novel loci, associated with choroidal thickness. Findings suggest inflammatory and vascular processes drive choroidal thickness, with overlapping mechanisms shared with refractive error. Genome-wide independently significant SNPs accounted for 18.7% of the genetic variance in choroidal thickness. Mendelian randomisation analyses showed a causal effect of age-related macular degeneration on choroidal thickness, and suggest a bidirectional causal effect between choroidal thickness and primary angle-closure glaucoma. These findings provide insight into the shared genetic architecture and biological pathways linking choroidal thickness and related diseases.

<h4>BACKGROUND</h4> Dementia with Lewy bodies shares clinical and pathological features with both Parkinson’s disease and Alzheimer’s disease, but the local biological factors that render specific cortical regions vulnerable to atrophy remain poorly defined. In particular, it is unclear whether cortical thinning in dementia with Lewy bodies reflects generic neurodegenerative mechanisms, processes shared with Parkinson’s disease and Alzheimer’s disease, or dementia with Lewy bodies-specific molecular and network susceptibilities. <h4>METHODS</h4> A total of 89 patients with dementia with Lewy bodies and 89 matched controls underwent T1-weighted brain MRI. Scans were processed to generate surface-based cortical thickness maps. Regional cortical thickness estimates, after slice-by-slice manual correction, were mapped to gene expression data from healthy postmortem human brains to identify transcriptomic signatures associated with decreased thickness in dementia with Lewy bodies. We assessed whether genes whose expression was increased with regional thinning converged onto established Parkinson’s disease- and Alzheimer’s disease-related pathways and isolated genes uniquely implicated in dementia with Lewy bodies. Spatial annotation mapping was then used to test whether patterns of cortical thinning overlapped with in vivo neurotransmitter system distributions and whether the observed thickness pattern was constrained by large-scale structural connectivity, consistent with a network-based propagation process. <h4>RESULTS</h4> Cortical thinning predominated in regions that, in the healthy brain, show higher expression of genes involved in mitochondrial function and synaptic transmission. The transcriptomic profile associated with thinning significantly overlapped with genes belonging to Parkinson’s disease and Alzheimer’s disease pathways, supporting shared pathogenic mechanisms across Lewy body and Alzheimer-type neurodegeneration. However, 90 genes associated with cortical thinning did not overlap with Parkinson’s disease or Alzheimer’s disease pathways and were enriched for GABAergic signalling. Spatial mapping analyses showed that regions with greatest thickness reductions colocalized with GABA A , serotoninergic 5-HT 1A , 5-HT 1B , 5-HT4, and dopaminergic D2 receptor distributions, and that the thickness pattern followed structural connectivity. <h4>CONCLUSIONS</h4> MRI-derived cortical thickness changes in dementia with Lewy bodies reflect selective molecular and network vulnerabilities rather than a non-specific degenerative process. Mitochondrial and synaptic genes, together with a distinct GABAergic association and connectivity constraints, delineate mechanisms explaining why some cortical territories are more affected in dementia with Lewy bodies.

Influenza A viruses (IAVs) cause severe outbreaks with high mortality in birds and humans. A deeper understanding of cell-intrinsic defense mechanisms against influenza viruses is therefore crucial for developing novel antiviral strategies. Herein, we perform a genome-wide CRISPR activation screen to systematically elucidate host restriction factors against influenza A (H7N9) virus. Among multiple candidates, cholesterol 25-hydroxylase (CH25H) is shown to be induced by influenza virus infection and inhibit viral membrane fusion. Notably, our previous work demonstrated that CH25H blocks the entry of plasma membrane-fusing viruses such as coronaviruses. This inhibition occurs by relocating accessible cholesterol from the plasma membrane (PM) to the endoplasmic reticulum (ER). Here, we extend this finding and show that the same mechanism works against endocytosis-dependent viruses such as influenza viruses. The exogenous supplementation of cholesterol can restore depleted accessible cholesterol and reverse the CH25H-mediated restriction. Additionally, we prove that acyl-CoA:cholesterol acyltransferase (ACAT) is required to recruit the accessible cholesterol in this process. However, how hydrophobic accessible cholesterol is transported remains unclear. Here, we demonstrate that GRAMD1/Aster-mediated non-vesicular cholesterol transport is utilized to mobilize accessible cholesterol upon stimulation of CH25H. 25-hydroxycholesterol (25HC), the catalytic product of CH25H, is a natural metabolite that potently inhibits influenza virus infection both <i>in vitro</i> and <i>in vivo</i>. These findings underscore the promising therapeutic potential of 25HC against influenza viruses.

The human genome reference established a shared coordinate system for genome function, but it is incomplete and not fully representative of human diversity. Here, we benchmark how genome representation and corresponding analytical frameworks for each representation shape functional genomics using chromatin accessibility sequencing (ATAC-seq), RNA sequencing, whole-genome bisulfite sequencing, and chromosome conformation capture (Hi-C) data from lymphoblastoid cell lines derived from five individuals with fully phased genome assemblies. We compare results across hg38, CHM13, the draft human pangenome, and each individual's maternal and paternal assemblies. Because current pipelines and quality control conventions are tuned to hg38, several of these comparisons reflect genome representation in the context of available methods, rather than sequence alone. Individual identity accounts for 57.52-78.47% of total variance in functional estimates, whereas genome choice contributes 0.002-7.85% and sample-by-genome interactions contribute 0.63-5.43%. About 2% of biological signals are detectable only with personal assemblies. Although these effects are modest overall, some biologically important features remain inaccessible to linear references. Consistent with this, graph-based DNA methylation analysis in the human pangenome reveals a non-reference AluY5a insertion within a putative TNKS enhancer at chromosome 8p23.1 that becomes visible and hypermethylated only in the pangenome.

<h4>Background</h4>Huntington's disease is caused by a trinucleotide CAG repeat expansion in the HTT gene. Despite displaying autosomal dominance, phenotypic variation exists amongst mutation carriers, in particular relating to the age that symptoms first occur. This variation is primarily driven by an inverse relationship between CAG expansion size and age of symptom onset. However, the majority of variation in age of onset that is independent of CAG repeat length is thought to be driven by environmental influences. Since DNA methylation can be altered by environmental factors, and as methylomic variation is reported in other neurodegenerative diseases, it may offer a potential mechanism underlying disease manifestation.<h4>Results</h4>We utilized the Illumina EPIC v1 methylation array to profile DNA methylation in 120 samples, including three distinct brain regions (striatum, entorhinal cortex and cerebellum) in 20 Huntington's disease and 22 control donors. We identified seven Bonferroni-significant differentially methylated CpGs within the striatum along with 27 differentially methylated regions, annotated to genes involved in physiological processes known to be disrupted in HD such as the urea cycle and metabolism. Weighted gene correlation network analysis identified modules of co-methylated CpGs that were associated with Huntington's disease, with ontological analyses showing enrichment in disease relevant processes. Furthermore, integration of single-nuclei RNA sequencing data highlighted that genes annotated to these modules are enriched in striatal spiny projection neurons, the primary cell types affected in the disease.<h4>Conclusions</h4>Here, we present the first epigenome-wide association study of Huntington's disease conducted in the striatum, the primary region of neuropathology, along with matched entorhinal cortex and cerebellum on the Illumina EPIC v1 array. Our results suggest that DNA methylation is altered at loci associated with Huntington's disease in disease relevant regions and cell types and strengthens evidence for areas of potential therapeutic intervention.

Huntington's disease is caused by a CAG expansion in the HTT gene, leading to somatic repeat instability, alternative processing of HTT pre-mRNA, and mutant huntingtin protein production. To model these features, we generated a knock-in minipig (KI-85Q-HD) carrying a (CAG)82CAA(CAG)2 repeat in the endogenous HTT locus. To evaluate this, we quantified somatic expansion in various tissues using small pool- and bulk-PCR; detected HTT1a, an aberrantly spliced HTT transcript, using 3' rapid amplification of cDNA ends and quantitative PCR; and assessed mutant huntingtin protein isoforms using homogeneous time-resolved fluorescence assays. Moderate levels of tissue-specific and age-dependent somatic expansion were observed, highest in the caudate nucleus, kidney and spleen, and detectable in blood cells. We confirmed the presence of HTT1a transcripts terminating at a cryptic polyadenylation site in HTT intron 1, and detected soluble full-length mutant HTT and HTT1a proteins across brain regions and peripheral tissues, while aggregated HTT1a was only detected in the cortex. These results indicate that KI-85Q-HD minipigs exhibit molecular features of Huntington's disease at a pre-symptomatic stage and may serve as a platform for assessing therapeutic distribution and potency.

No abstract available.

Acute myeloid leukemia (AML) harboring KMT2A rearrangement (KMT2Ar) was generally associated with poor prognosis. We enrolled 490 patients with KMT2Ar from the East China Leukemia Alliance between March 2013 and August 2025. KMT2A::MLLT3 was the most frequent KMT2Ar (31.6%), followed by KMT2A::MLLT4 (25.1%), KMT2A::ELL (20%), and KMT2A::MLLT10 (12.7%). KMT2A::MLLT4 showed an inferior prognosis. The most co-occurring gene mutations were KRAS (20.1%), NRAS (19.0%), TET2 (10.2%), WT1 (8.6%), and PTPN11 (7.4%). Trisomy 8 was more common in patients < 60 years, while FLT3-ITD was only detected in patients < 60 years. With a median follow-up time of 42.6 months, the median overall survival (OS) of all patients was 30.9 months, and the 3-year OS rate was 49.9%. Patients treated with venetoclax plus intensive chemotherapy (Ven+IC) showed the best composite complete remission (CRc) rate and OS compared to intensive chemotherapy, venetoclax plus reduced-intensive chemotherapy, and reduced-intensive chemotherapy (CRc rate: 89.5% vs. 62.4% vs. 57.3% vs. 61.4%; median OS: not reached vs. 39.9 months vs. 34.8 months vs. 12.7 months). Multivariate analysis identified multiparameter flow cytometry minimal residual disease negativity post-induction therapy, allogeneic hematopoietic stem cell transplantation, and Ven+IC as independent favorable prognostic factors for event-free survival (EFS), and KMT2A::MLLT4 fusion, EVI1 overexpression were independent unfavorable prognostic factors for EFS. In summary, our study showed characteristics and prognostic implications in newly diagnosed KMT2Ar AML in China.

Phosphorus fixation represents a primary constraint limiting the agronomic efficiency of phosphate fertilizers in calcareous soil. Rapeseed roots secrete amounts of organic matter, which can mobilize and decompose insoluble phosphorus in the soil. However, the activation mechanism of sparingly insoluble phosphorus in calcareous soils by rapeseed rhizosphere microorganisms remains unclear. This study aimed to screen and identify phosphate-solubilizing microorganisms from the rapeseed rhizosphere of calcareous soil, and to elucidate their key metabolic pathways for activating insoluble phosphorus. The results demonstrated that (i) fourteen dominant phosphate-solubilizing strains were isolated from rapeseed rhizosphere soil. Among these, <i>Advenella alkanexedens</i> was verified to significantly promote wheat growth and increase soil available phosphorus content. (ii) The culture condition optimization and functional characterization for <i>Advenella alkanexedens</i> revealed that its optimal growth temperature was 30°C, with an initial pH of 7. Its phosphate-solubilizing ability was regulated by Mg²⁺, K²⁺, and Ca²⁺ ions, and the strain exhibited considerable salt tolerance and the ability to produce siderophores. (iii) <i>Advenella alkanexedens</i> increased soil available phosphorus content by 11.49%-81.91% and elevated phytase activity by 36.87%-82.49%. Correlation analysis indicated that soil available phosphorus and phytase activity were significantly positively correlated with Ca₂-P, Ca₈-P, and Al-P fractions. (iv) Amino acids and organic acids were identified as the key metabolites influencing the phosphate-solubilizing function of <i>Advenella alkanexedens</i>. The KEGG pathway analysis showed these metabolites were primarily enriched in β-alanine metabolism and arginine and proline metabolism pathways. Our findings confirm that <i>Advenella alkanexedens</i> not only promotes crop growth but also significantly increases labile P fractions (Ca₂-P, Ca₈-P, Al-P) while reducing more stable forms (Ca₁₀-P), thereby enhancing soil phosphorus use efficiency. This study holds important implications for planting rapeseed to activate insoluble phosphorus in soil, to reduce phosphate fertilizer application, and to promote sustainable utilization of soil phosphorus resources. Furthermore, it provides a theoretical foundation for developing agricultural microbial inoculants.IMPORTANCEOur results confirm that <i>Advenella alkanexedens</i> not only benefits crop growth but also converts insoluble phosphates (O-P, Ca10-P) into highly active inorganic phosphorus components, thereby enhancing the utilization efficiency of soil phosphorus. This study was of great significance in activating the insoluble phosphorus in the soil, reducing the input of phosphate fertilizers, achieving the sustainable utilization of phosphorus resources, and protecting the environment. Additionally, it provided a basis for developing agricultural microbial agents.

<h4>Purpose</h4>Anemia, inflammation and iron deficiency are linked to Fibroblast growth factor 23 (FGF23). Aim of this study was to explore the dynamics of FGF23 in Hereditary Hemochromatosis type-I (HH1).<h4>Methods</h4>Twenty-six consecutive patients with genetically confirmed and uncomplicated HH1 and nineteen healthy age-matched voluntary blood donors (CTR) were enrolled for the study. Intact (iFGF23) and C-terminal (cFGF23) FGF23 and iron status markers were evaluated at baseline (T0) and seven days (T7) after phlebotomy/voluntary blood donation (VBD). Bone mineral density (BMD) and vertebral fracture assessment (VFA) were also evaluated at T0.<h4>Results</h4>Cross-sectional and longitudinal analyses failed to reveal significant differences in iFGF23 in both HH1 (T0: 54.27 ± 14.42 pg/mL; T7: 54.70 ± 15.48 pg/mL) and CTR (T0: 52.77 ± 17.63 pg/mL; T7: 53.27 ± 15.88 pg/mL) groups. Absence of significant difference was also observed for cFGF23 at baseline between the two groups (HH1, T0: 0.98 ± 0.39 pmol/L; CTR, T0: 1.18 ± 0.91 pmol/L) but not at T7 when the values (HH1, T7: 01.20 ± 0.89; CTR, T7: 1.84 ± 1.11 pmol/L) were significantly increased in CTR compared to T0 (p = 0.0003) and to HH1 (p = 0.0240). After phlebotomy/VBD, in both HH1 and CTR groups, serum levels of phosphate were unchanged from baseline while serum iron, ferritin and transferrin saturation and erythrocyte-related parameters (red blood cells, hemoglobin and hematocrit) were significantly reduced (all p  <  0.0001). Serum iron, ferritin, and transferrin saturation were significantly higher in HH1 than in CTR (p = 0.0002 for serum iron and p  <  0.0001 for both ferritin and transferrin saturation) at T7. In the CTR group, these parameters showed a trend toward iron deficiency whereas in HH1 they remained near the upper limit of the normal range, suggesting a persistent mild iron overload. Correlation and regression analyses did not show significant associations between circulating FGF23 (both iFGF23 and cFGF23) and iron status markers. BMD and VFA were not significantly different between HH1 and CTR. Furthermore, BMD and Trabecular Bone Score values were not associated with circulating FGF23 levels.<h4>Conclusion</h4>This pilot study indicates that the serum levels of iFGF23 and cFGF23 and skeletal health evaluated through BMD and VFA do not differ between patients with uncomplicated HH1 and healthy subjects at baseline. The absence of changes in the serum levels of iFGF23 and cFGF23 in HH1 patients with uncomplicated HH1 after phlebotomy may reflect the persistence of iron overload which could counteract the physiological hypoxia-driven stimulation of FGF23 production and cleavage observed in healthy subjects after VBD.

Differentiating autism spectrum disorder (ASD) from developmental delay (DD) is critical for guiding early intervention, but overlapping features and shared biological mechanisms pose challenges. This study investigates whether copy number variations (CNVs) affecting serotonergic genes carry sufficient information to distinguish between these neurodevelopmental disorders (NDDs). Using network mapping and machine learning, we applied gene ontology (GO) terms related to serotonergic systems to filter CNVs and construct networks modeling genetic and biological variation in ASD and DD. We identified hub nodes and subnetworks reflecting distinct patterns in gene and GO term interactions. ASD networks analysis yielded six genetic clusters, five of which remarkably contained genes specifically linked to serotonergic receptor mechanisms. In contrast, DD networks exhibited greater genetic homogeneity, with just two clusters sharing serotonergic mechanisms. Random Forest classifiers using serotonergic gene features achieved an average prediction accuracy of 85.6%, increasing to 88.6% when combined with dopaminergic dosage features, consistent with the two systems capturing partially non-overlapping biological signal. GO-based features yielded comparable accuracy with fewer inputs, emphasizing their efficiency. These findings demonstrate that different genetic alterations may be associated with disruption of shared biological pathways, each leaving a distinct signature tied to clinical diagnoses. Importantly, although genetic heterogeneity is observed in ASD, we found a homogeneity of serotonergic biological terms, suggesting convergence of mechanisms, which was distinct for each condition. Together, these results suggest that serotonergic CNVs carry discriminatory information for ASD vs. DD classification, and that combining serotonergic and dopaminergic features captures partially non-overlapping biological signal.

Haematopoietic stem cells (HSCs) produce all blood and immune cells throughout life, but ageing progressively impairs their function, generating excessive myeloid and megakaryocyte cells at the expense of lymphocytes. This lineage imbalance contributes to immune decline, chronic inflammation and increased disease susceptibility in the elderly, yet the underlying mechanisms remain poorly understood. Here we show that a specific Meg3⁺ HSC subset (CD150^hiSca1^hiCD24^hiCD201⁺CD9⁺CD63⁺ long-term HSCs) expands dramatically during ageing and drives this lineage skewing. Using multi-omics profiling, we found that inflammatory signals increase H3K23ac levels in aged Meg3⁺ HSCs, enhancing PU.1 activity through recruitment of the reader protein TRIM24. This epigenetic mechanism promotes excessive megakaryocyte and myeloid production. Of note, disrupting H3K23ac-TRIM24 interaction in aged HSCs restored balanced lineage output and reduced inflammatory signals. Our findings reveal a key mechanism linking inflammation to HSC ageing and identify potential therapeutic targets for reversing ageing-related immune decline.

Histone modifiers are crucial for instructing multiple-stage cellular differentiation, yet the mechanisms underlying their temporal precision remain enigmatic. Here, we demonstrate that the H3K27 demethylase Kdm6b acts as an epigenetic regulator, coordinating stepwise motor neuron (MN) differentiation through sequential partnerships with stage-specific transcription factors (TFs). Genome-wide profiling reveals a progressive gain in Kdm6b occupancy, especially at distal regulatory elements, as differentiation proceeds. Kdm6b dynamically remodels chromatin landscapes by coordinating H3K27me3 removal with H3K27ac and H3K4me1 acquisition, thereby enabling timed gene activation from MN specification to maturation. Stage-specific inhibition of Kdm6b compromises the ordered expression of developmental genes. Mechanistically, Kdm6b interacts with temporal TFs over time to ensure precise transcriptional control and MN differentiation. Our work elucidates how a single epigenetic regulator achieves temporal fidelity of stepwise MN development, providing insight into epigenetic regulation of developmental timing.

Viral polymerases are key targets for antiviral drug development due to their crucial role in viral replication. This study seeks to discover novel inhibitors using virtual screening of a 200,583-member ZINC-based library, targeting three aspartyl-based viral polymerases: SARS-CoV-2 RdRp, HIV-1 RT, and HCV NS5B. In this line, applying RO5, drug-likeness, and PAINS filters, leading to a pharmacophore model with Ribavirin as the benchmark. Four ligands (Ligands 1-4) with the highest binding affinities were identified via molecular docking and ADME/T studies. These candidates were then subjected to 200 ns molecular dynamics (MD) simulations and binding free-energy calculations to evaluate binding stability. The computational analyses demonstrated that all four ligands stably bound the polymerase active sites and induced some conformational changes. According to MM/PBSA results, Lig-2 and Lig-1 surpass Ribavirin in SARS-CoV-2 RdRp and HIV-1 RT complexes, while Ribavirin showed the strongest binding in HCV NS5B. ADME/T studies supported the pharmacological potential of the ligands, with Lig-2 emerging as a promising lead against all targets, and Lig-1, Lig-3, and Lig-4 showing potential for multi-target antiviral development.

Transcriptional dysregulation is among the most prominent molecular alterations in Huntington's disease (HD). It is not confined to the brain but also extends to peripheral tissues and cells, enabling minimally invasive screening strategies to identify transcriptional surrogates of the health status in HD mutation carriers. Nonetheless, transcriptomics approaches have failed to identify consistent candidates from peripheral blood, probably due to the low impact of the HD mutation in the transcriptional profiles of circulating cells, which can be masked by the high cellular complexity of this biofluid. In this study, we applied for the first time single-cell RNA-seq to peripheral blood mononuclear cells (PBMCs) to determine which cells accumulate the most prominent gene expression changes, and therefore, represent potential sources of reliable biomarkers. We observed common transcriptional alterations across different blood cell subtypes, which were partially validated in published bulk transcriptomics datasets. To relate these gene expression patterns with disease progression in the absence of a large cohort of patients, we examined selected candidates in a phenotypically characterized cohort of transgenic R6/1 mice. Among the tested genes, only the variations in the interferon related gene Irf7 in blood were mildly correlated with motor coordination performance in mutant mice. Notably, striatal Irf7 expression did not show such phenotypical correlation in the same individuals. Overall, transcriptional-based changes in peripheral blood can be linked to HD but they are mild and not apparently confined to particular cellular subpopulations. In conclusion, dissection of individual blood cells in combination with subsequent validation in mouse models emphasizes the challenges in obtaining clinically relevant biomarkers in HD.

Endoplasmic reticulum autophagy (ER-phagy) is a selective autophagy pathway in which receptor proteins target ER membranes and proteins for degradation, yet its role in Alzheimer's disease (AD) remains unclear. Here, we identify FAM134B/RETREG1 as a specific ER-phagy receptor mediating amyloid precursor protein (APP) degradation. FAM134B directly interacts with ER-localized wild-type and familial mutant APP via their C-terminal domains and recruits LC3 through its LC3-interacting region (LIR) to promote APP delivery to phagophores for lysosomal degradation. In AD, epigenetic silencing at the FAM134B promoter suppresses its transcription by limiting TFEB/TFE3 binding despite their nuclear enrichment. This transcriptional suppression impairs ER-phagy, leading to APP accumulation and exacerbated AD pathology. AAV-mediated hippocampal expression of wild-type, but not LIR-mutant, FAM134B in 5XFAD mice restores ER-phagy, enhances APP clearance, reduces Aβ deposition, preserves synaptic and myelin integrity, and improves cognitive performance. These findings establish FAM134B downregulation as an upstream pathogenic event in AD, suggesting ER-phagy enhancement as a promising strategy to suppress Aβ generation at its source.

Humic acid (HA) is known to improve phosphorus (P) availability in agricultural soils. Yet, the underlying mechanisms by which it influences the microbial community and subsequent P turnover remain unclear. In this study, a pot experiment was conducted using soil from a three-year Salvia miltiorrhiza (S. miltiorrhiza) cultivation system, in which the soil was amended with three HA concentrations (T1: 100-fold dilution, T2: 200-fold, T3: 400-fold) alongside an untreated control (CK). We determined P uptake by S. miltiorrhiza, soil P fractions, phosphatase activities, along with high-throughput sequencing of the microbial communities to specifically target those associated with P transformation. The results showed that HA application significantly enhanced root P uptake, with increases of 68.59% and 91.05% under T2 and T3, respectively. Consequently, soil Olsen-P content decreased by 19.19% and 15.20%, respectively, consistent with the depletion of available P under enhanced plant uptake. Soil P fractionation further revealed that HA application decreased inorganic P by 20.91% and 32.29%, respectively, under T2 and T3 treatments. Specifically, H₂O-P decreased by 62.1% and 73.61%, and NaHCO₃-Pi decreased by 53.21% and 50.48%, respectively, under the T2 and T3 treatments. In parallel, acid phosphatase activity was increased by 68.72% and 66.67% under the T2 and T3 treatments compared to CK. Comparative high-throughput sequencing between T2 and CK revealed that HA application enriched key microbial genera associated with P cycling, including Sphingomonas, Nitrospira, Ferruginibacter, and Hyphomicrobium. Collectively, these findings suggest that HA may promote P mobilization and mineralization through association with microbial communities, thereby potentially enhancing P bioavailability and plant uptake. These findings offer new perspectives on the associations between HA application, microbial community shifts, and P use efficiency, suggesting a potential approach that merits further evaluation for P management in agricultural systems.

<h4>Background</h4>Antithrombin deficiency is a rare hereditary predisposition to venous thromboembolism caused by variants in the SERPINC1 gene. In up to 20% of cases, no explanatory variant is identified.<h4>Objectives</h4>This study uncovered novel variants by covering the full SERPINC1 region.<h4>Methods</h4>In this single-center cohort study, we included antithrombin-deficient families in which no SERPINC1 variant had been previously identified. We used multiple genomic and transcriptomic approaches to identify novel variants and assess their pathogenicity. Variants were classified according to SERPINC1-specific American College of Medical Genetics and Genomics criteria.<h4>Results</h4>We included 10 families with 42 individuals and identified 3 novel SERPINC1 (NM_000488.4) variants: c.42-209T>G, c.1153+429C>T, and c.916T>G. In addition, we identified an exon 6 deletion of uncertain novelty. All variants cosegregated perfectly. Despite being highly suspect, c.42-209T>G was the only variant of uncertain significance according to American College of Medical Genetics and Genomics criteria, which prompted us to perform functional testing. Prediction tools suggested the introduction of an acceptor splice site at c.42-208, resulting in the inclusion of a pseudoexon with premature stopcodon. We confirmed this aberrant splicing prediction by quantitative polymerase chain reaction and Sanger sequencing of cDNA synthesized from RNA isolated from genomic-edited HepG2 cells. c.42-209T>G leads to a reduction of SERPINC1 transcript levels.<h4>Conclusion</h4>We report several novel SERPINC1 variants, one of which is the first pathogenic deep-intronic variant causing antithrombin deficiency through aberrant splicing. This finding improves our understanding of the genetic etiology and clinical practice for antithrombin deficiency. In all tested families registered with our institution, an explanatory SERPINC1 variant has now been identified.

<i>Xylaria grammica</i> is an endophytic fungus that produces grammicin, a polyketide reported as a structural isomer of the mycotoxin patulin. However, the environmental regulation of grammicin production remains unclear, and the full biosynthetic pathway has not yet been completely resolved. Genome re-annotation and biosynthetic gene cluster mining revealed high secondary metabolite potential, with 92 predicted clusters spanning major biosynthetic classes. Time-series RNA sequencing under shaking dark, static dark, and continuous light conditions showed strong condition-dependent transcriptome divergence. Light-grown cultures were enriched in functions associated with stress adaptation and detoxification, including oxidoreductase- and monooxygenase-related categories, whereas static dark cultures showed broader activation of secondary metabolism-associated genes at later stages. Among the predicted biosynthetic gene clusters, a patulin-like locus in <i>X. grammica</i> contained homologs corresponding to most genes of the <i>Aspergillus clavatus pat</i> cluster from <i>PatA</i> to <i>PatO</i>, although the gene order differed. Expression profiling revealed coordinated induction of patulin-like genes after 3 days post-inoculation under dark conditions, whereas continuous light strongly repressed their transcription. Quantitative RT-PCR analysis further confirmed this darkness-dependent activation pattern. Together, with previous functional evidence showing that <i>patK</i> and <i>patL</i> are required for grammicin production, these results indicate that darkness acts as a major regulatory switch for secondary metabolism in <i>X. grammica</i> and support the involvement of a patulin-like locus in grammicin biosynthesis. However, the downstream tailoring steps leading to grammicin remain unresolved, and further genetic and metabolomic analyses will be required to complete pathway reconstruction.

All-inorganic CsPbBr₃ perovskite solar cells (PSCs) suffer from efficiency losses due to their interfacial energy level mismatch. Herein, we propose a synergistic dual-interface modification strategy to improve the photoelectrical performance of C/CsPbBr₃/SnO₂-structure PSCs. At the CsPbBr₃/carbon interface, <i>N</i>,<i>N</i>'-dicyclohexylcarbodiimide (DCC) could elevate the CsPbBr₃ Fermi level to suppress the interfacial non-radiative recombination to increase the open-circuit voltage (<i>V</i> _OC). At the SnO₂/CsPbBr₃ interface, NH₄F could upshift the SnO₂ Fermi level to enhance the crystallinity of CsPbBr₃ to markedly improve the fill factor (FF). The optimized device could achieve a power conversion efficiency of 9.59% with a <i>V</i> _OC of 1.53 V and an FF of 82.4%. To elucidate the underlying mechanism, we established a dual-diode series model (DDSM), which revealed that the <i>V</i> _OC was governed by the CsPbBr₃/carbon barrier and the FF was controlled by the SnO₂/CsPbBr₃ barrier. The theoretical predictions were consistent with the experimental results, providing a robust framework for designing high-performance all-inorganic PSCs.

The development of sustainable, economical sensing platforms for rapid monitoring of toxic metals is a critical priority for environmental safety. In this work, a new dual-functional BNP@RHNS nanocomposite sensor was fabricated by immobilizing Br-naphthophen ligand onto agrowaste (rice husk)-derived mesoporous nanosilica. The sensor surface maintains the amorphous nanosilica framework and successfully anchors ligands covalently or non-covalently, according to the physicochemical characterization methods used. The sensor exhibited remarkable spectrophotometric selectivity toward Ag⁺ and Fe³⁺ ions in aqueous environments with an optimized response time <40 s. The method also showed broad linear dynamic ranges of 0-0.8 ppm and 0-0.85 ppm, and low limits of detection (LOD) of 0.185 ppm and 0.247 ppm for Ag⁺ and Fe³⁺ ions under the optimized conditions (pH 6.2 & pH 3.6 at 25 °C). The sensing mechanism was meticulously detailed through experimental spectral shifts and corroborated by Density Functional Theory (DFT) calculations. The theoretical work showed that the recognition process is governed by the spontaneous formation of stable coordinate complexes located in the ligand's N₂O₂ cavity. Moreover, the sensor exhibited high interfering tolerance and was successfully applied to determining metal ions in real water matrices with satisfactory recovery rates (97.37% to 100.73%). Furthermore, this sensor was found to be effective in recovering Ag(i) from leachates of electronic waste, suggesting its broader applicability in urban mining and resource recovery. These findings suggest a promising alternative valorization strategy for rice husk biomass into high-value functional materials for advanced environmental monitoring applications.

Autism spectrum disorder (ASD) may be a predisposing factor in the development of catatonia, which has been demonstrated to be effectively treated using electroconvulsive therapy (ECT). CACNA1 channelopathy mutations have been associated with autism spectrum disorder, intellectual disability, and epilepsy. The single published case report of ECT use in CACNA1 subunit mutations demonstrated a negative outcome, and no current case reports on CACNA1B, CACNA1C, CACNA1D, or CACNA1E mutations have been published. We present a case of an adolescent with a known CACNA1D mutation and symptoms of excited catatonia who experienced a reduction in the frequency and severity of self-injurious and violent behavior following treatment with ECT. Our case demonstrates the use of ECT for catatonia in an individual with a CACNA1 mutation without complication and with a positive therapeutic response.

<h4>Objectives</h4>Breast cancer (BC) subtypes such as HR+, HER2+, and triple-negative (TNBC) show distinct molecular features, treatment responses, and outcomes. DNA methylation is a key, targetable epigenetic regulator in BC. This study examined whether the DNA methyltransferase inhibitor decitabine (DAC) produces subtype-specific epigenomic and transcriptional effects in breast cancer cell lines representing distinct molecular subtypes.<h4>Methods</h4>Gene expression and DNA methylation data from DAC-treated and untreated T-47D (Luminal-A) and JIMT-1 (HER2-amplified, trastuzumab-resistant with a TNBC-like phenotype) breast cancer cell lines were obtained from a published dataset. Differential expressions were assessed using <i>limma</i>, and methylation changes were defined using β-value thresholds. Integrated epigenomic-transcriptional analysis, functional enrichment, Horvath clock CpG evaluation, and survival analysis were performed in the METABRIC and TCGA cohorts.<h4>Results</h4>In JIMT-1, DAC caused hypomethylation at 1195 CpG sites and upregulation of 187 genes, including <i>TFAP2E</i>, an age-associated locus selectively hypomethylated after DAC. In T-47D, DAC induced hypomethylation at 1937 CpGs and upregulated 248 genes. Amongst these, <i>KRT20</i> was upregulated despite promoter hypermethylation, indicating a subtype-specific regulatory architecture. DAC-responsive genes in JIMT-1 were enriched for cytokine signaling and piRNA-mediated epigenetic silencing, whereas T-47D showed enrichment for extracellular matrix organization, collagen dynamics, and piRNA processing pathways. Horvath clock CpG analysis showed selective perturbation of age-associated sites. Survival analysis identified 114 DAC-responsive genes associated with overall survival in ER/PR-positive BC and 8 in the JIMT-1-derived gene set.<h4>Conclusion</h4>DAC induces subtype-dependent epigenomic and transcriptional remodeling, selectively disrupts age-associated regulatory programs, and underscores the need for subtype-stratified evaluation of epigenetic therapies in breast cancer.

As the primary gatekeeper of ferroptosis, glutathione peroxidase 4 (GPX4) represents a compelling therapeutic target. A recent study demonstrates that sustained GPX4 blockade via sophisticated lipidic nanoplatforms simultaneously triggers robust ferroptosis and remodels the tumor immune microenvironment; however, the underlying synergistic mechanisms remain poorly elucidated. By integrating classical GPX4 enzymatic theories with recent breakthroughs regarding its membrane-bound structural functions and its secretion as an immune-disrupting ligand, we propose a framework illustrating how GPX4 serves as a nexus between intracellular survival mechanisms and extracellular immune evasion. More importantly, we delineate a "spatial topological" landscape of GPX4 to fully harness its therapeutic potential in cancer therapy.

Neuroinflammation is a fundamental pathological hallmark driving the initiation and progression of various neurological disorders. Luteolin, a natural flavonoid abundant in medicinal plants, fruits, and vegetables, exerts multifaceted neuroprotective effects across diverse experimental disease models. Its beneficial activities are mediated through complementary mechanisms, including suppression of aberrant microglial activation, modulation of pro-inflammatory signaling pathways, and enhancement of endogenous antioxidant defenses. These integrated actions attenuate inflammatory mediator release, reduce oxidative stress-induced neuronal damage, and inhibit apoptosis, thereby counteracting neuroinflammation-driven pathology. This review synthesizes current knowledge on luteolin's protective roles in central nervous system (CNS) disorders. It elucidates underlying molecular mechanisms, encompassing regulation of key signaling cascades such as NF-κB, MAPK, and Nrf2, as well as its impact on cellular processes including autophagy and mitochondrial function. Critical challenges hindering clinical translation-notably limited oral bioavailability and restricted blood-brain barrier (BBB) permeability-are systematically discussed to guide future research. A comprehensive understanding of luteolin's pleiotropic pharmacological actions will not only enhance knowledge of its therapeutic potential but may also facilitate the development of novel preventive and therapeutic strategies for neuroinflammatory and neurodegenerative diseases.

<h4>Introduction</h4>We leveraged consanguinity and population endogamy in a large four generation Turkish family with ET. Examination of clinical features and genetic analysis identified a homozygous <i>PRPF40B</i> missense variant, in a large region of homozygosity, segregating with ET in the family.<h4>Methods</h4>The ET family is of Turkish origin. The proband and relatives were evaluated at Ankara University Medical School and Bilkent University (Ankara, Turkey). A second evaluation of the clinical and videotape data was performed at Yale University. A total of 6 individuals from the family were clinically assessed. Whole genome sequencing and autozygosity mapping was performed in affected and unaffected family members. Transient transfection of HEK293T cells was performed to assess pathogenicity of the <i>PRPF40B</i> missense variant.<h4>Results</h4>A total of 4 family members were diagnosed with ET; two individuals also had a diagnosis of Parkinson's Disease (PD). We identified a missense variant in <i>PRPF40B</i>, p.Ser166Leu, located within a 7.7 Mb region of homozygosity (ROH) (Chr12: 46595701-54,330,441), which was observed in all analyzed affected family members. Unlike wildtype <i>PRPF40B</i> which localizes to nuclear speckles in the nuclear compartment, when mutant <i>PRPF40B</i> was expressed in HEK293T cells we observed enrichment and localization to the nuclear membrane.<h4>Discussion</h4>The <i>PRPF40B</i> p.Ser166Leu variant is located within a conserved region of the WW protein domain. <i>In Silico</i> pathogenicity and functional studies predict that the missense variant alters the function of <i>PRPF40B</i>. <i>PRPF40B</i> has previously been implicated in the pathogenesis of neurological disorders, including Huntington's disease and Rett syndrome.

<h4>Introduction</h4>Microglia, the resident immune cells of the central nervous system, play a critical role in maintaining neural homeostasis and regulating inflammatory responses in the brain. Increasing evidence suggests that microglial dysfunction contributes to the progression of neurodegenerative diseases, including Alzheimer's disease (AD). However, the molecular mechanisms underlying these alterations remain incompletely understood. This study aimed to characterize disease-associated molecular changes in microglia derived from induced pluripotent stem cells (iPSCs) of sporadic AD patients and healthy donors.<h4>Methods</h4>iPSC-derived microglia from sporadic AD patients and healthy controls were analyzed using integrated multi-omics approaches, including total RNA sequencing, proteomics, and small non-coding RNA (sncRNA) sequencing. Gene Ontology (GO) analysis was performed to identify dysregulated biological pathways from transcriptomic and proteomic datasets. In addition, a modified T4 polynucleotide kinase (T4 PNK)-based sncRNA sequencing method was used to profile disease-associated sncRNAs and identify previously uncharacterized RNA species.<h4>Results</h4>Comparative analyses revealed significant AD-associated alterations in mRNA, protein, and sncRNA expression profiles in iPSC-derived microglia. GO analysis demonstrated dysregulation of pathways related to extracellular communication, intracellular transport, cytoskeletal organization, and protein-protein interactions. Furthermore, the modified T4 PNK-sncRNA sequencing approach identified multiple disease-associated sncRNAs, including several novel and previously uncharacterized RNA species potentially linked to AD pathology.<h4>Discussion</h4>These findings demonstrate that iPSC-derived microglia provide a valuable model for studying molecular mechanisms associated with sporadic AD. The identified transcriptomic, proteomic, and sncRNA alterations highlight key pathways potentially involved in microglial dysfunction and neurodegeneration. In particular, the discovery of novel disease-associated sncRNAs may provide new insights into AD pathogenesis and reveal potential therapeutic targets for future investigation.

<h4>Introduction</h4>GABAergic interneurons (IN) are critical for the precise timing and flow of information in cortical circuits. Loss of GABAergic IN function has been suggested as a potential translationally relevant mechanism of neuropathology in Fragile X Syndrome (FXS). Indeed, in rodent models of FXS, some IN populations may display reduced number, while genes associated with other IN type upregulated. However, it remains unknown how these cell populations, and their cell-type specific gene expression patterns are regulated in early development across other mammalian models of FXS.<h4>Methods</h4>Here we utilise an outbred rat model of FXS, in which we have performed single-nucleus RNA sequencing analysis in neonatal development of the somatosensory cortex. We then use immunohistochemistry to measure the number and distribution of neurochemically identified GABAergic IN subtypes from early development until adolescence in the somatosensory cortex.<h4>Results</h4>We find that GABAergic INs in a rat model of FXS display clear evidence of transcriptomic alteration compared to wild-type littermates in early brain development. These effects are most profound in putative parvalbumin INs, but with modest changes in other cell types. From immunohistochemistry, we find that parvalbumin INs appear largely unaffected in density in distribution, but we observe a large upregulation in the number of somatostatin-expressing INs.<h4>Conclusion</h4>GABAergic INs may display cell-type specific transcriptomic regulation in response to the loss of FMRP. Our data suggests minimal alteration of parvalbumin IN density or distribution, but upregulated somatostatin cell numbers. These data partially agree with previous observations in mouse models of FXS.

Inflammation is an essential host defense mechanism but, when excessive or chronic, can cause severe tissue damage and organ dysfunction. Exosomes, a key subtype of extracellular vesicles, have emerged as critical mediators of intercellular communication by transferring bioactive cargo and modulating inflammatory signaling. This review summarizes the roles of exosomes in inflammation-associated tissue injury, focusing on their regulation of classical inflammatory pathways and programmed cell death. We further discuss the dual functions of exosomes as pathogenic mediators and therapeutic agents, highlighting their diagnostic and therapeutic potential.

Oral squamous cell carcinoma, the most prevalent malignancy of the head and neck, presents ongoing challenges regarding its molecular mechanisms and clinical management. Current research largely focuses on isolated signaling pathways or specific immune responses, often overlooking the potential contributing role of the microbiota in tumorigenesis. This review proposes the "microbiome-inflammation-immune axis" as an interpretive working hypothesis to elucidate how dysbiosis and microbial interactions may activate host inflammatory responses, trigger pattern recognition receptors and signaling pathways, and remodel the immune microenvironment, thereby potentially facilitating oral cancer progression. Concurrently, this paper emphasizes clinical translation by critically compiling and evaluating relevant clinical analytical indicators-such as peripheral blood inflammatory markers and salivary microbial markers-from the fields of inflammation and microbiology. This provides multidimensional reference points for future disease diagnosis and treatment.

Diabetic kidney disease (DKD) is a leading cause of end-stage renal disease, with renal fibrosis as its core pathological hallmark. A central driver of this fibrosis is epithelial-mesenchymal transition (EMT), during which renal tubular epithelial cells transform into matrix-producing myofibroblasts. Endothelial-mesenchymal transition (EndMT) has also emerged as a critical contributor, and together with EMT, accounts for the progressive accumulation of myofibroblasts and extracellular matrix. A major clinical challenge in halting DKD progression is "metabolic memory", a phenomenon whereby renal injury persists and EMT/EndMT remain activated even after glycemic control is achieved. The molecular basis underlying this sustained activation remains incompletely understood. Emerging evidence indicates that metabolic memory is largely mediated by epigenetic mechanisms, including histone modifications, DNA methylation, and non-coding RNA dysregulation. These stable epigenetic imprints maintain the persistent activation of key pro-fibrotic signaling pathways, especially TGF-β, thereby continuously driving EMT, EndMT, and excessive extracellular matrix deposition. Although targeting epigenetic regulators has shown promising anti-fibrotic effects, a systematic review that integrates how metabolic memory orchestrates both EMT and EndMT through a multi-layered epigenetic network remains lacking. This review comprehensively summarizes the epigenetic mechanisms by which metabolic memory sustains EMT and EndMT in DKD, highlights key therapeutic targets, and discusses their translational and clinical implications.

The Fontan procedure is a palliative operation for patients with univentricular physiology. Liver complications post-surgery are inevitable and include liver cirrhosis and hepatocellular carcinoma (HCC). Routine surveillance includes transient elastography (TE) and clinical assessment, however longitudinal data on Fontan-associated liver disease (FALD) is scarce. This study aimed to describe the evolution of FALD and assess the correlation between liver stiffness measurement (LSM) and hepatic decompensation. This retrospective case series comprised all adult post-Fontan patients presenting to a tertiary hospital between January 2015 and December 2024. Demographic and clinical information, serum tests, TE and echocardiograms were extracted from medical records. Endpoints assessed were decompensated cirrhosis and findings that warrant HCC multi-disciplinary team review. A derived surrogate outcome, clinically significant portal hypertension (CSPH) was assessed using the Baveno VII criteria. There were a total of 65 patients with 111 Fibroscans™. There was no significant change in LSM by TE when comparing two consecutive decade years post Fontan surgery. On echocardiography, only inferior vena-cava diameter and degree of ventricular function correlated with LSM scores (p = 0.032, p < 0.001). Twelve patients had CSPH, correlating with increased gamma-glutamyltransferase, bilirubin, international normalised ratio and aspartate aminotransferase levels (p = 0.047, 0.034, 0.012, 0.049 respectively). Four patients exhibited features of decompensated cirrhosis, eight warranted multi-disciplinary team discussion however these did not correlate with LSM or steatosis results. LSM and echocardiogram monitoring alone is insufficient for identifying FALD patients at risk of hepatic decompensation. Regular testing and multicenter research is required to develop predictive models that accurately identifies these at-risk patients.

Left ventricular hypertrophy (LVH) is a not uncommon finding on cardiac imaging, seen in 10-20% of adults. LVH can represent a response to a physiological load on the heart (either by athletic training, hypertension, or structural heart issues) or it can represent a primary pathology (e.g. infiltrative disorders, including cardiac amyloidosis and sarcoidosis) or familial/genetic conditions (e.g. hypertrophic cardiomyopathy). LVH is an independent risk factor for adverse cardiovascular events, regardless of its etiology. In this review, we survey the etiologies, clinical approach, and use of multimodality imaging in those with LVH.

<i>Background and Objectives</i>: MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression and have emerged as potential biomarkers in type 2 diabetes mellitus and its complications. This pilot exploratory study aimed to identify circulating miRNAs with differential expression in plasma from patients with newly diagnosed type 2 diabetes mellitus compared to age- and sex-matched healthy controls. <i>Materials and Methods</i>: Peripheral venous blood samples were collected from diabetic patients (<i>n</i> = 24) and controls (<i>n</i> = 12). Due to the exploratory nature of the study and limited sample material, samples were pooled within each group prior to plasma separation. Total RNA, including miRNAs, was extracted from plasma and analyzed using a high-throughput qPCR panel. Two normalization methods were applied to assess miRNA expression, and overlapping results were used for downstream analysis. Fold regulation was calculated using the 2^(-ΔCt) method. <i>Results</i>: A total of 33 and 42 miRNAs were identified as differentially expressed using the first and second normalization methods, respectively. Fourteen miRNAs were consistently downregulated across both methods. Several of these miRNAs, including hsa-miR-26a-5p, hsa-miR-146a-5p, hsa-miR-186-5p, hsa-miR-19a-3p, and hsa-miR-652-3p, have been previously associated with glucose metabolism, inflammation, and diabetic complications, such as retinopathy, neuropathy, and endothelial dysfunction. The pooling strategy enabled an efficient exploratory assessment of miRNA expression patterns while reducing inter-individual variability. <i>Conclusions</i>: This exploratory pilot study identifies a panel of circulating miRNAs with altered expression in pooled plasma samples from patients with newly diagnosed type 2 diabetes mellitus. These findings provide preliminary insights that warrant further validation in larger, individual-level studies to assess their diagnostic and prognostic potential.

<h4>Purpose</h4>Despite improvements in and access to clinical genetic testing, many patients with classic phenotypes remain without a molecular diagnosis. Such cases may reflect variants in noncoding regions or complex alleles made up of disparate sequence variants. In this article, we report a patient with autosomal dominant polycystic kidney disease caused by a deep intronic de novo single-nucleotide substitution forming a dinucleotide variant, with an adjacent common single-nucleotide variant (formerly single-nucleotide polymorphism) inherited from an unaffected parent. Although neither variant alone was predicted to be pathogenic, together as a dinucleotide variant, they created a novel splice donor site that activated a pseudoexon within intron 16 of the <i>PKD1</i> gene.<h4>Methods</h4>Genome sequencing was paired with transcriptome sequencing to evaluate the dinucleotide variant within intron 16 of <i>PKD1</i>.<h4>Results</h4>Evaluation of RNA from patient cells confirmed the inclusion of a 114-bp pseudoexon and an in-frame premature termination codon into the <i>PKD1</i> messenger RNA transcript. This case identified a rare scenario in which the combination of 2 independently predicted benign nucleotide substitutions immediately adjacent to one another generated a pathogenic allele.<h4>Conclusion</h4>This case highlights the diagnostic utility of genome sequencing coupled with RNA sequencing to clarify the effects of a deep intronic variant, confirming a molecular diagnosis for a patient with a clear clinical diagnosis who had undergone years of inconclusive testing. Furthermore, this case cautions that similar events may underlie unsolved cases across other genes and disorders.

No abstract available.

Developmental and epileptic encephalopathies (DEEs) are a group of neurological disorders primarily affecting young children and are characterized by severe seizures. DEEs are challenging to manage, with some patients experiencing severe side effects or not responding to frontline therapies. This is partly because of the many underlying mechanisms involved in DEE pathology and the relatively limited mechanism-specific action of current treatments. The CACNA1E gene, which encodes the voltage-gated calcium channel Cav2.3 (R-type), has recently been associated with DEEs. More than fifteen different mutations in CACNA1E have been identified in patients with DEEs; however, the mechanisms by which these mutations affect channel function and, thus, their relationship to DEEs, remain largely unknown. Previous research has begun to characterize the functional effects of R-type channel mutations on channel biophysics, but only a handful of mutations have been studied functionally to date. Here, we transiently expressed Cav2.3 channels and used whole-cell patch-clamp to examine the biophysics of one specific disease-associated R-type channel mutant in which leucine 228 is substituted with a proline (L228P). Compared to wild-type, the L228P mutant did not alter peak current density, inactivation kinetics, or recovery from inactivation, but showed a significant shift towards hyperpolarized voltages in both voltage-dependent activation and steady-state inactivation. This resulted in a broader window current shifted towards more hyperpolarized potentials, which predicts increased channel availability and activity at subthreshold voltages relative to wild-type channels. Our results contribute to the ongoing characterization of R-type mutants, with the long-term goal of informing mechanism-specific therapies for DEEs.

<h4>Background</h4>Adolescent idiopathic scoliosis (AIS) is a complex spinal deformity with evidence of contributions from multiple genetic factors, but the molecular basis of gene-gene interactions in its development remains unclear. Filamin B (FLNB) and tetratricopeptide repeat domain 26 (TTC26) have both been implicated in the regulation of primary cilia and extracellular matrix homeostasis. However, whether combined variation in these genes contributes to spinal instability has not been established.<h4>Results</h4>We generated mice carrying heterozygous mutations in both Flnb and Ttc26 and find that these double heterozygous mice develop progressive spinal curvature during puberty, whereas single gene mutants show only mild susceptibility. We observed reduced production of collagen type II and aggrecan in the intervertebral disc, accompanied by increased expression of matrix degrading enzymes. Transcriptomic and cellular analyses show that simultaneous loss of Flnb and Ttc26 suppresses Sonic hedgehog (Shh) signaling in the nucleus pulposus, due to impaired methylation and nuclear accumulation of the transcription factor GLI2. We find that TTC26 is required for the localization of protein arginine methyltransferase 7 (PRMT7) to the primary cilium, enabling methylation of GLI2, while FLNB binds methylated GLI2 to promote its nuclear import. Restoring Shh signaling in vivo improves extracellular matrix composition in mutant discs.<h4>Conclusions</h4>These findings show that FLNB and TTC26 act together to regulate Shh-GLI2 signaling and maintain extracellular matrix homeostasis in the intervertebral disc. Their combined disruption compromises spine stability and may contribute to the pathogenesis of adolescent idiopathic scoliosis.

<h4>Background</h4>Working memory deficits are common in mood disorders and severely affect everyday functioning. Serotonin (5-HT) signalling has been implicated in depression and is also involved in cognitive functioning. However, its relevance for working memory remains largely unexplored.<h4>Aims</h4>Using positron emission tomography (PET) brain imaging, we investigated the link between working memory and multiple 5-HT brain features in both healthy individuals and patients with mood disorders in a cross-sectional analysis of pooled data-sets.<h4>Method</h4>We used multiple linear regression to test the associations between working memory performance and 5-HT 1B receptor (5-HT_1BR) (healthy controls: 28), 5-HT 2A receptor (5-HT_2AR) (healthy controls: 116), 5-HT 4 receptor (5-HT₄R) (healthy controls: 97, patients: 98) and 5-HT transporter (5-HTT) (healthy controls: 137, patients: 12) PET binding in the frontal cortex. The frontal cortex was chosen as region of interest as it is critical for working memory functions.<h4>Results</h4>There was no association between working memory and 5-HT_1BR (<i>p</i> = 0.14), 5-HT_2AR (<i>p</i> = 0.99) or 5-HTT (<i>p</i> = 0.80) frontal cortex binding in healthy controls. For the 5-HT₄R, we observed a significant interaction effect of group status (<i>p</i> = 0.01), with patients showing a positive association (<i>β</i> = 6.51, <i>p</i> = 0.02) and healthy individuals showing no significant association (<i>p</i> = 0.16).<h4>Conclusions</h4>We found no evidence that key 5-HT receptor systems are associated with working memory performance in healthy individuals, but did observe a positive association for 5-HT₄R in patients with mood disorder. We speculate that although 5-HT neurotransmission markers may map onto working memory performance in the healthy state, pathologically altered 5-HT signalling may contribute to working memory deficits in mood disorders, possibly through downstream signalling and/or interactions with other neurotransmitter systems.

The co-occurrence of skeletal and neuropsychiatric disorders suggests underlying bone-brain associations, yet their characterization remains limited by the anatomical and biological complexity of both systems. Here, we systematically examine these associations across diverse anatomical locations using structural imaging and genetic data from approximately 45,000 UK Biobank participants. We identify widespread structural associations between bone and brain, with 34.9% of examined pairs showing significant relationships. These associations display pronounced regional heterogeneity, with effect size variation aligning with gradients of cortical development and evolutionary expansion, as well as spatial distributions of GABA and 5HT-1b receptors and gene expression profiles of oligodendrocyte precursor cells and astrocytes. Genetic analyses spanning genome-wide, pathway, locus, and single-variant levels further reveal shared genetic architecture between skeletal and brain structures, as well as with multiple neuropsychiatric and neurological traits. Enrichment within the Wnt signaling pathway highlights a potential mechanistic axis underlying bone-brain coupling, while distinct skeletal regions exhibit differential genetic links to specific brain-related health outcomes. Together, these findings demonstrate that bone-brain associations are pervasive yet exhibit substantial anatomical and genetic heterogeneity, providing a systems-level framework for understanding the shared biology of skeletal and brain health.

The dorsal striatum is important for highly specialized functions including movement, learning, and habit formation. However, it is not known if species-specialized behaviors are associated with cellular specializations in the striatum. Here, we compared single-nucleus RNA sequencing (snRNA-seq) data from human, chimpanzee, rhesus macaque, common marmoset, and pale spear-nosed bat caudate (CN) and putamen (Pu) separately as well as mouse caudoputamen (C-Pu), which represents divergence among species spanning approximately 94 million years of evolution. We observed a lower neuron-to-glia ratio in primate striata compared to non-primates, reflecting the allometric scaling of neuron density and relative glia density invariance in larger brains. Among neurons, eccentric spiny projection neurons (eSPNs) - an SPN of unknown function - showed significantly lower proportions in non-primate striata for both CN and Pu. Focusing on the heterogeneity within interneurons, we identified two bat striatal interneuron cell types that are nearly absent in other species: which express LMO3, and co-express FOXP2 and TSHZ2. Other striatal interneurons also exhibited differential abundance between primates and non-primates. In summary, we provide a comprehensive snRNA-seq dataset of dorsal striatum, identify two distinct, previously uncharacterized populations of bat interneurons, and uncover fundamental cellular composition differences between primate and non-primate striata.

Excited-state intramolecular proton transfer (ESIPT) plays a critical role in many chemical processes, and it is essential to develop diverse regulating mechanisms of ESIPT molecules. Herein we report control of ESIPT luminogens with the combination of competing dynamic covalent bonds and non-covalent hydrogen bonds. The incorporation of amino-containing ESIPT fluorophores into 2-formylbenzenesulfonamide offers a versatile platform for dictating reactivity and fluorescence. The competition between intramolecular hydrogen bonding of sulfonamide NH and dynamic ring-fusion of NH with nearby aldehyde site in ring-chain tautomers enables diverse emission of cyclic hemiaminal, ketoimine upon ESIPT, and anionic intramolecular charge transfer (ICT) band in solution. Dynamic covalent reactions with amines give luminogen-dependent ketoimine or ICT emission, with hydrogen bonding from imine nitrogen affecting proton transfer of sulfonamide. In the solid state of one luminogen the engagement of adjacent aldehyde for intramolecular hydrogen bonding affords ICT emission while higher energy excitation induced ketoimine emission. Mechanical grinding and acid fuming treatment allows structural conversion and cyclic hemiaminal/ketoimine emission, further achieving switch of multistate emission colors. Finally, dynamic covalent reactions with amino acids provide a facile way for tuning distinct emission in the solution and solid state. The results should find broad utility in molecular switches, chemical sensing, and intelligent materials.

Vitamin D receptor (VDR) agonists promote quiescence of cancer-associated fibroblasts and improve efficacy of chemotherapy in preclinical models of pancreatic cancer. We conducted a run-in phase trial with primary endpoint of safety when the VDR agonist paricalcitol is given with first-line gemcitabine and albumin-bound paclitaxel (GA) in patients with metastatic pancreatic cancer. Secondary endpoints included pharmacodynamic analyses. Thirty-six patients were randomized to GA plus placebo, GA plus intravenous paricalcitol or GA plus oral paricalcitol with pretreatment and on-treatment tumor biopsies. Paricalcitol was safely administered with GA, although five patients (42%) receiving oral paricalcitol had grade 2-4 hypercalcemia and required dose reduction. Nuclear VDR protein expression was heterogeneous across patients, and VDR was expressed in tumor, immune and stromal cells. Compared to pretreatment specimens, on-treatment biopsies had decreased proportion of αSMA⁺ fibroblasts, altered fibroblast VDR activation signature and greater density and spatial colocalization of CD8⁺ T cells with tumor cells in the GA-plus-paricalcitol arms. VDR expression was predictive of tumor response in the GA-plus-paricalcitol arms. Paricalcitol can be safely administered with chemotherapy to patients with metastatic pancreatic cancer, and on-treatment biopsies indicated favorable modulation of the tumor microenvironment by paricalcitol as predicted by preclinical models. ClinicalTrials.gov identifier: NCT03520790 .

Crohn's disease (CD) is highly heterogeneous in presentation and progression with no cure. Molecular phenotyping has been used to elucidate cellular and tissue-based alterations to characterize drivers and effects of disease. One currently understudied class of functional molecules is long non-coding RNAs (lncRNAs). Studying the full lncRNA landscape in CD is challenging due in part to an incomplete lncRNA annotation and a lack of their functional characterization in tissues of interest. We used a genome-guided alignment strategy to assemble predicted lncRNA transcripts using short RNA-sequencing data from colon tissue of adult patient samples. When combining our predicted lncRNAs with previous lncRNA annotations, we determined 98 that were differentially expressed, recapitulating many from previous IBD studies while also uncovering new ones. We built gene co-expression networks to cluster lncRNAs with functionally characterized protein-coding genes. Clusters containing differential lncRNAs were correlated to disease status and associated with pathways related to the humoral immune response, metabolism, and tissue regeneration. We uncovered multiple differential lncRNAs whose expression significantly correlated with nearby differential protein-coding genes that have also been differentially expressed in other IBD datasets, such as PITX2. We focused on a predicted lncRNA that is antisense to the PITX2-adjacent lncRNA PANCR, which we called PANCR-AS1, and provide multiple lines of evidence that support PANCR-AS1 functioning as an enhancer of PITX2 expression. Overall, we determined lncRNAs that are potential contributors to CD pathogenesis. We developed a robust pipeline for identifying lncRNAs in diseased and non-diseased tissue that are absent from reference annotations. We also outlined a framework to pinpoint significant disease-associated lncRNAs with potential functional activity related to their nearby protein-coding genes.

<h4>Background</h4>Global olive cultivation is often hindered by the physiological phenomenon known as alternate bearing, which results in inconsistent yields from year to year, thereby diminishing economic returns and challenging effective orchard management. While foliar nutrition is acknowledged as a viable approach to boost reproductive performance in olive trees, there is limited research exploring the combined use of NPK and calcium boron nano fertilizers (NPK-NF and CaB-NF) during the 'off-year' season to concurrently enhance flowering, fruit set, oil composition, and reduce the severity of alternate bearing.<h4>Results</h4>Field experiments conducted during the 2023 and 2024 seasons on fifteen-year-old 'Picual' olive trees demonstrated that foliar application of NPK-NF and CaB-NF significantly enhanced reproductive performance and oil quality while reducing alternate bearing severity. Trees receiving NPK-NF (4 mL/L) + CaB-NF (2 mL/L) exhibited maximum flowering parameters, including 9.31 inflorescences per shoot and 68.51% perfect flowers, alongside the highest initial fruit set (37.37%) and final fruit set (26.95%). This treatment produced the greatest 'off-year' yield (0.79 tonnes per acre) while reducing fruit drop to 27.83% compared to 51.77% in controls. Leaf and fruit mineral analysis revealed substantially enhanced nitrogen, phosphorus, potassium, and calcium concentrations in treated trees. Virgin olive oil maintained extra virgin classification with improved oleic acid proportions, reaching 67.85%, and increased pigment concentrations. Notably, the combination of NPK-NF (4 mL/L) + CaB-NF (3 mL/L) reduced alternate bearing severity to 31.06% compared to 88.08% in control trees.<h4>Conclusion</h4>These findings suggest that the deliberate foliar application of NPK-NF and CaB-NF during the 'off-year' season can significantly boost flowering, fruit set, and oil quality, while also alleviating the severity of alternate bearing in 'Picual' olive cultivar. This approach offers a viable nutritional strategy to enhance both yield stability and productivity in commercial olive orchards.

<h4>Background</h4>This study aimed to evaluate the surgical outcomes of parathyroidectomy (PTX) for secondary hyperparathyroidism (SHPT) in hemodialysis patients, comparing the use of three operative techniques: high-frequency electrosurgery (HFE), ultrasonic scalpel (US), and bipolar electrocoagulation (BPE).<h4>Methods</h4>The patients were divided into three groups based on the type of surgical instruments used. Outcomes assessed included surgical efficiency, postoperative complications, recurrence rate, hospitalization costs, biochemical and nutritional parameters over 1 year, and overall survival through March 2025.<h4>Results</h4>A total of 171 patients were retrospectively categorized into three groups: HFE (n = 50), US (n = 60), and BPE (n = 61). Compared to group HFE, both the US and BPE groups exhibited significantly shorter durations for drainage tube removal, operative times, and postoperative hospital stays (all p < 0.05). The BPE group also had significantly lower intraoperative blood loss compared to the US and HFE groups (p < 0.05) and incurred the lowest hospitalization costs. The incidence of postoperative complications and recurrence rates did not differ significantly among groups. All groups showed marked reductions in serum calcium, phosphorus, and intact parathyroid hormone levels postoperatively, which stabilized over time with no significant differences between groups. At the 1-year follow-up, patients exhibited significant improvements in anemia-related and nutritional markers compared to baseline. With follow-up extending to March 2025, overall survival did not differ significantly among the three surgical groups (p = 0.987).<h4>Conclusions</h4>PTX effectively corrected SHPT, improved anemia, and enhanced nutritional status in hemodialysis patients. All three surgical techniques, HFE, US, and BPE, were safe and effective, with comparable long-term survival. However, US and BPE offered superior operative efficiency, and BPE represented the most cost-effective option.

Gestational diabetes mellitus (GDM) is one of the most common complications during pregnancy and can severely impair fetal neurodevelopment, leading to abnormalities in emotional, cognitive, and memory functions. However, the precise mechanisms by which GDM affects fetal neural development remain incompletely understood, which is closely associated with the lack of animal models that accurately replicate the key clinical and pathological features of human GDM. Therefore, this review summarizes and evaluates current methodologies for establishing GDM animal models, elucidates the impact of GDM on offspring neurodevelopment and its potential underlying mechanisms, and provides a theoretical foundation for developing and assessing appropriate GDM animal models, clarifying the pathophysiological mechanisms of GDM, and exploring innovative therapeutic strategies.

<h4>Background</h4>Traditionally viewed as a localized "wear-and-tear" pathology, osteoarthritis (OA) is now increasingly recognized as a complex systemic disorder driven by metabolic and inflammatory dysregulation. This review synthesizes emerging evidence to redefine the pathogenesis of OA from a "whole-joint" to a "whole-body" perspective.<h4>Main body</h4>We first examine local degradation mechanisms, identifying synovial macrophage polarization, mitochondrial dysfunction, and autophagy defects as critical drivers of a pro-inflammatory milieu. Furthermore, we elucidate the mechanism of inflammatory "spillover," wherein intra-articular cytokines (e.g. IL-1β, TNF-α) and extracellular vesicles (EVs) enter the circulation, contributing to a state of low-grade systemic inflammation. This systemic inflammatory burden is closely associated with a cascade of comorbidities, including endothelial dysfunction and atherosclerosis potentially mediated by shared mechanisms such as the "bone-vascular axis," sarcopenia through the pain-disuse cycle, and central sensitization coupled with HPA axis dysregulation. Conversely, systemic metabolic disorders, particularly obesity-induced "metaflammation" and insulin resistance, exacerbate joint degeneration through adipokines (e.g. leptin, resistin), forming a vicious bidirectional cycle.<h4>Conclusions</h4>We conclude by discussing how this systemic paradigm necessitates a shift in therapeutic strategies, moving from symptomatic management to holistic interventions. These include targeting metabolic pathways (e.g. metformin), clearing senescent cells (senolytics), and adopting a multidisciplinary precision medicine approach based on inflammatory and metabolic phenotyping.

<h4>Background</h4>Average daily gain (ADG), age at 115 kg (AGE), backfat thickness (BF), and loin eye area (LEA) are key growth and carcass traits in pigs. However, breed-specific molecular regulatory mechanisms underlying these traits remain largely unknown. Here, we identified multiple candidate genes associated with these 4 traits from Duroc and Yorkshire pigs by three Genome-wide association study(GWAS) methods (MLM, FarmCPU, and BLINK) and differential genes expression analysis, verified them based on RNA sequencing(RNA-seq), Assay for Transposase-Accessible Chromatin using sequencing (ATAC-seq), and Chromatin Immunoprecipitation sequencing (ChIP-seq) data from Duroc and Yorkshire pigs, and explored the regulatory mechanisms underlying these traits.<h4>Result</h4>GWAS across Duroc, Landrace, and Yorkshire identified 44 high-confidence SNPs. A total of 24, 13, and 19 candidate genes were identified respectively from Duroc, Landrace, and Yorkshire by GWAS and differential gene expression analysis (in three breeds versus Meishan pigs). The candidate gene ARL8A was found to be highly conserved between Duroc and Yorkshire. GeneMANIA networks of Landrace exhibited tightly connected core modules of candidate genes. Integrated RNA-seq, ATAC-seq, and ChIP-seq analyses of muscle and fat tissue data consistently suggested that ARL8A exhibits relatively high transcriptional activity and chromatin accessibility, supporting its potential role in muscle and fat development.<h4>Conclusions</h4>The integration of three GWAS methods with differential gene expression analysis revealed that core candidate genes involved were mainly involved in regulating fat deposition, energy metabolism, and cell growth in constructed breed-specific regulatory networks. Among these candidate genes, ARL8A was a highly conserved gene modulating growth-related traits (BF and LEA), as evidenced by the multi-omics data. Our findings provide an insight into genetic architecture and regulation mechanisms underlying pig growth and development, offering important resources for porcine molecular breeding and genetic improvement.

<h4>Background</h4>Lipid metabolic reprogramming is increasingly recognized as a critical feature of prostate cancer progression, but the lipid metabolism-related genes that remain continuously dysregulated from normal tissue to primary tumor and metastatic disease have not been systematically characterized, and their biological and prognostic relevance remains incompletely understood.<h4>Objective</h4>To identify lipid metabolism-related genes associated with continuous prostate cancer progression and develop a prognostic signature for survival stratification.<h4>Methods</h4>Clinical prostate cancer specimens and a high-fat diet (HFD)-driven RM-1 tumor model were first used to evaluate lipid metabolic alterations in vivo. GSE6919 transcriptomic data were used to identify genes shared between the Normal-Primary and Primary-Metastatic transitions. These genes were intersected with a curated lipid metabolism-related gene set, followed by GO and KEGG enrichment analyses. TCGA prostate adenocarcinoma expression and clinical data were used for LASSO regression to construct a prognostic model. The four core genes were further evaluated by clinicopathological correlation analysis, protein- and transcript-level validation in clinical tissues and prostate cancer cell lines, and functional assays under oleic acid-induced lipid stress. Immune infiltration analysis, ssGSEA, and nomogram analysis were performed to assess the biological and clinical relevance of the model.<h4>Results</h4>Clinical tissues showed increased PLIN3 expression, and HFD feeding promoted tumor growth and reinforced lipid metabolic alterations in vivo. A total of 44 lipid metabolism-related genes were identified as continuously dysregulated during prostate cancer progression. These genes were mainly enriched in fatty acid metabolism, lipid catabolism, peroxisome, lipid droplet, glycolysis/gluconeogenesis, arachidonic acid metabolism, and PPAR signaling. Eight genes were significantly associated with overall survival in TCGA, and a four-gene signature comprising ALDH3A2, ENO2, PPP1CB, and PTGIS was established. This model effectively stratified patients into high- and low-risk groups with significantly different survival outcomes. The risk score was positively associated with clinical T stage and Gleason score. The four core genes were also associated with lipid metabolic enzymes, immune infiltration patterns, and multiple metabolism-related pathways. Protein- and transcript-level validation in clinical tissues and prostate cancer cell lines supported the biological relevance of the signature, although PTGIS showed a more context-dependent pattern. Functionally, silencing ENO2 reduced oleic acid-induced lipid peroxidation, whereas silencing PPP1CB enhanced it, while ALDH3A2 showed a more context-dependent effect. A nomogram integrating the risk score with clinical variables improved individualized survival prediction.<h4>Conclusion</h4>We identified lipid metabolism-related genes continuously dysregulated during prostate cancer progression and established a four-gene prognostic signature with potential value for survival prediction and risk assessment. These findings highlight lipid metabolic rewiring as an important component of prostate cancer evolution and provide candidate biomarkers for future mechanistic and translational studies.

The ketogenic diet (KD) is increasingly recognized as a promising therapeutic strategy for neurodegenerative disorders because of its multifaceted impacts on key pathophysiological mechanisms. This review explores the molecular pathways through which KD may protect against neurodegeneration, including the use of ketone bodies as alternative energy substrates, reduction of oxidative stress and inflammation, modulation of autophagy and protein aggregation, and impact on the gut microbiome. The potential benefits of KD are explored across neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, and multiple sclerosis, based on both preclinical and clinical evidence that supports its feasibility. However, challenges in long-term safety, patient adherence, and clinical practicality limit its widespread adoption. This review underscores the potential of KD for treating neurodegeneration on the basis of current scientific evidence while highlighting the need for further research to optimize its application and address existing gaps.

Male infertility is a complex and heterogeneous disorder. It has a significant genetic component, although some cases remain idiopathic. Next-generation sequencing, particularly whole-exome (WES) and whole-genome sequencing (WGS) has become a powerful tool for uncovering new genetic causes. This systematic review aimed to identify and synthesize genes linked to male infertility reported in WES or WGS studies. The review was registered on PROSPERO (CRD42024597301). A bibliographic search was conducted on PubMed, Scopus, and Web of Science (2014-2024). We included human studies that used WES or WGS to investigate the genetics of male infertility. Two reviewers independently performed screening, data extraction, and risk-of-bias assessment with JBI checklists. Out of 8018 identified records, 23 studies met the inclusion criteria. In total, 169 unique genes were reported; after removing duplicates, 143 genes remained. The most frequently implicated phenotypes were multiple morphological abnormalities of the flagella (MMAF) and non-obstructive azoospermia (NOA). Genes were stratified by recurrence across independent cohorts and by functional validation status. Of the 143 genes, five were replicated with functional validation, 22 demonstrated either replication or functional validation, and 116 were reported in single studies with only in silico support. In studies applying the criteria of American College of Medical Genetics and Genomics (ACMG), the diagnostic yield was 48% for MMAF and 12-23% for NOA, with an average variant-of-uncertain-significance (VUS) burden of 47%. ACMG classification was inconsistently applied across studies. MMAF-associated genes were predominantly autosomal recessive (95%), whereas NOA-associated genes exhibited greater diversity: 60% autosomal recessive, 25% X-linked, and 15% autosomal dominant. Only 34% of genes had undergone functional validation. In summary, among WES and WGS studies of predominantly sporadic cohorts, most identified genes were reported in single studies without functional validation, and standardized variant classification was implemented in only a minority of studies.

No abstract available.

Colorectal liver metastasis (CRLM) remains the primary cause of mortality in patients with colorectal cancer (CRC), yet effective predictive tools and reliable biomarkers are still lacking. DeepMetabio-mCRC Screener, an integrated multi-omics framework combining large-scale transcriptomic profiles with serum metabolomics, was developed to address this gap. In a cohort of 1,077 CRC samples, 620 metabolism-related genes were used to train a convolutional neural network, yielding an area under the receiver operating characteristic curve of 0.92 in the validation cohort and 0.97 in the independent testing cohort, outperforming the performance of the 10 established machine learning models. Model-derived transcriptomic risk scores revealed 22 core metabolic features associated with metastatic progression and CRLM occurrence, particularly retinol and tryptophan metabolism. Cross-omics integration revealed aminocarboxymuconate-semialdehyde decarboxylase (ACMSD) as a promising biomarker associated with impaired nicotinamide adenine dinucleotide biosynthesis. Clinical validation in 100 CRC patients confirmed elevated ACMSD levels in patients with CRLM, which correlated with advanced stage, recurrence risk, an immune-inflamed tumor microenvironment, and heightened sensitivity to epidermal growth factor receptor/vascular endothelial growth factor receptor-targeted therapies. In vitro, ACMSD knockdown was associated not only with suppressed CRC cell migration caused by inhibition of the transforming growth factor-β/epithelial-to-mesenchymal transition pathway but also with decreased proinflammatory and immune-responsive pathways and reduced immune cell infiltration. These findings collectively validate the DeepMetabio-mCRC Screener as a substantial early risk prediction tool and underscore ACMSD, identified through this framework, as a multifunctional biomarker for diagnosis, prognosis, molecular characterization, and therapeutic decision-making in patients with CRLM.

Triptolide is a bioactive diterpenoid lactone from traditional Chinese medicine with notable anti-tumor, immunosuppressive, and anti-inflammatory properties. However, its clinical utility is hampered by poor solubility, a narrow therapeutic index, and multi-organ toxicity. In response, extensive efforts have been made to improve its efficacy and safety profile. This review critically examines recent progress in Triptolide research, covering its mechanisms of action, derivative design, drug delivery systems, and cancer-specific applications. It also evaluates its clinical potential against pancreatic, breast, and ovarian cancers, addresses ongoing challenges, and suggests future research paths to accelerate clinical translation. This article presents a narrative review summarizing the research progress on the pharmacological activities, structural modifications, and nanodelivery systems of Triptolide. Literature searches were conducted in PubMed and Web of Science using keywords including "Triptolide", "pharmacological activity", "structural modification", and "nanodelivery", combined with Boolean operators. The search timeframe was from database inception to March 2026. Inclusion criteria were: original research articles related to the pharmacological activities, structural modifications or nanodelivery systems of Triptolide, peer-reviewed journal articles published in English or Chinese. Exclusion criteria were: duplicate publications, conference abstracts, dissertations, and preprints, articles irrelevant to the topic. Study selection was performed independently by two reviewers, with disagreements resolved through discussion or consultation with a third reviewer. A total of 158 articles were included in this review.

Lactylation (Kla) is a novel post-translation protein modification regulation mechanism realized by the covalent coupling of lactate generated by glycolysis and lysine residues. Lactylation primarily includes histone Kla and non-histone Kla. Kla participates in biological processes in cells by regulating gene transcription and protein expression, as well as subcellular localization. Multiple organs are exposed to lactate metabolism disorder due to aging. Recent studies have demonstrated that Kla mediated by lactate metabolism disorder is involved in the occurrence and development of various age-related diseases. This article summarizes the functions of Kla in age-related diseases and the related mechanisms.

<h4>Purpose of review</h4>Modern oncologic therapies, including immune checkpoint inhibitors (ICIs), targeted kinase inhibitors (TKIs), antibody-drug conjugates (ADCs), bispecific antibodies, and adoptive cellular therapies, have transformed cancer care while introducing diverse cutaneous adverse events (cAEs). This review summarizes recent advances in the mechanistic understanding, clinical patterns, and management of dermatologic toxicities associated with contemporary cancer therapeutics.<h4>Recent findings</h4>Emerging evidence indicates that therapy-associated cAEs often reflect distinct biological mechanisms rather than nonspecific drug reactions. These toxicities can be conceptually organized into four major mechanistic paradigms: immune disinhibition, epithelial signaling inhibition, cytotoxic epithelial injury, and cytokine-driven immune activation. While immune checkpoint inhibitors remain the most extensively characterized model, newer therapeutic platforms, including ADCs and immune-engaging cellular therapies, have introduced additional toxicity patterns that are only beginning to be systematically characterized. Recent clinicopathologic studies have clarified cytotoxic epithelial injury patterns associated with ADCs, while early clinical series suggest cytokine-mediated inflammatory eruptions may occur during cellular immunotherapies. At the same time, advances in immunopathologic profiling have supported the development of mechanism-directed management strategies, including targeted cytokine blockade for steroid-refractory immune-mediated dermatoses.<h4>Summary</h4>Cutaneous adverse events associated with modern cancer therapies increasingly represent mechanism-based toxicities linked to therapy-specific biological pathways. Integrating clinical morphology with treatment class and immunologic mechanisms provides a practical framework for diagnosis and management. Mechanism-directed dermatologic care, including early recognition, targeted immunomodulation, and multidisciplinary collaboration, may improve toxicity control while preserving oncologic efficacy.

Tremor syndromes arise from diverse combinations of neuronal oscillation, resonance, and entrainment within interconnected cerebellar, thalamocortical, basal ganglia, and sensorimotor loops. This review synthesizes anatomical, physiological, pathological, clinical, and computational evidence to identify the most likely network mechanisms of common and uncommon tremor syndromes. Mechanical-reflex tremor reflects underdamped musculoskeletal resonance and reflex loop dynamics, whereas central neurogenic tremors arise from intrinsic membrane conductances, electrical coupling, and recurrent inhibitory and excitatory loops that support oscillation, frequency-selective resonance, and entrainment. Acute cerebellar lesions produce intention tremor through impaired dentate-mediated motor planning and interpositus-dependent predictive motor control, causing hypermetria and abnormal transcortical mechanical-reflex oscillation. Delayed tremor after a cerebellothalamic pathway injury reflects maladaptive plasticity, likely within the thalamocortical loop, and similar delayed maladaptive change plus abnormal basal ganglia oscillation produces Holmes tremor. Essential tremor probably originates from oscillatory cerebellar cortical circuits with thalamocortical amplification, shaped by heterogeneous genetic vulnerabilities. Parkinson tremor is produced by increased pallidal and subthalamic oscillation combined with thalamocortical resonance in response to a loss of dopaminergic modulation. Dystonic tremor arises from abnormal oscillation within interacting basal ganglia and cerebellar circuits. Cortical tremor stems from cortical hyperexcitability and reverberation. The properties of orthostatic tremor are most consistent with oscillation originating in the fastigium and associated cerebellar cortex with resonant amplification primarily in the pontobulbar reticular formation and secondarily in the thalamocortical loop. Neuropathic tremors appear to begin as disturbances of physiological mechanical-reflex and central neurogenic pathways but may evolve into disabling oscillation in the cerebellothalamocortical pathway. Tremor in sarcomeric myopathies probably emerges from unexplored central mechanisms of tremorogenesis. Future progress in tremor pathophysiology will require additional animal models and computational network models that can experimentally test proposed oscillatory mechanisms, distinguish causal oscillators from entrained amplifiers, and guide mechanism-based therapies.

<h4>Background</h4>Lynch Syndrome (LS) is an autosomal dominant disease characterized by germline heterozygous mutations in DNA mismatch repair (MMR) genes. High-risk LS patients may proceed to colorectal cancer (CRC). However, the drivers or biomarkers of LS benign colon tissue approaching malignant CRC are not completely understood. This study aimed to understand the molecular and cellular changes during malignant transition in LS.<h4>Methods</h4>Single-cell RNA sequencing (scRNA-seq) was used to analyze paired fresh biopsy samples from 3 LS patients (carcinoma vs. para-carcinoma, labeled as LS-CA vs. LS-paraCA). Single-nuclear RNA sequencing (snRNA-seq) was used to analyze a frozen biopsy sample of a LS patient. Datasets of Healthy controls and patients diagnosed with sporadic CRC (without LS-related germline or somatic mutations; labeled as nonLS-CRC) were downloaded from the open source. Integrative computational analysis was performed to conclude potential drivers of the malignant transition. Immuno-histo-fluorescence staining (IHF) were also performed for validating the proposed three key markers.<h4>Results</h4>In the single-cell atlas, we observed an increase of primitive cancer stem-cells with high expression of biomarkers <i>CEACAM5, BACE2</i>, <i>GPRC5A</i> and <i>OLFM4</i> in the epithelium of the LS. Both infiltration of immune cells and pathways related to DNA repair biological activity in LS are dramatically increased in carcinoma compared to para-carcinoma. The mutation burden in LS is fundamentally elevated compared to that in healthy controls. Furthermore, T cell and macrophage-related tumor immunity in LS is readily mobilized in carcinomas compared to para-carcinoma.<h4>Conclusions</h4>This study provides single-cell transcriptomic resource using affected tissues from patients with Lynch Syndrome and describes an integrative profile covering the alterations of cancer stem cell markers, mutation burden, and tumor immunity during the malignant transition from latency state to Lynch Syndrome and to colorectal cancer at the single-cell level.

Immunotherapy resistance in triple-negative breast cancer (TNBC) remains a critical clinical challenge. This review synthesizes and critically evaluates current evidence on two interconnected mechanisms that have emerged from recent research: the functional plasticity of mast cells (MCs) and metabolic reprogramming. Far from being mere allergic effectors, MCs exhibit high phenotypic diversity, with antigen-presenting MCs (apMCs) representing a unique subset identified in recent studies capable of priming anti-tumor T cell responses. A landmark phase 2 trial has demonstrated that modulation of apMCs can enhance responses to anti-PD-1 therapy in patients with ICI-refractory TNBC. However, within the TNBC TIME, metabolic dysregulation-characterized by glycolytic flux, lactate accumulation, and lipid alterations-skews MCs toward immunosuppressive phenotypes and suppresses apMC function. Published studies have documented that this bidirectional crosstalk forms a vicious cycle that sustains immune evasion and limits the efficacy of immune checkpoint inhibitors. To decipher this complexity, patient-derived organoid (PDO) models co-cultured with autologous immune cells have emerged as a validated platform that preserves tumor heterogeneity and enables real-time dissection of metabolism-immune circuits. This review systematically synthesizes current knowledge on the biological basis of MC plasticity, the metabolite-driven regulation of their function, and the utility of organoid-based systems for mechanistic discovery and drug screening. We critically evaluate emerging multidimensional therapeutic strategies, including pharmacological reprogramming of apMCs, metabolic normalization, and engineered cell therapies, and identify key knowledge gaps that must be addressed to translate these insights into clinical practice. By integrating advances in immuno-oncology, cancer metabolism, and bioengineering, this review provides a framework for translating current insights into clinically actionable strategies to overcome immunotherapy resistance in TNBC.

Aristolochic acid (AA), a naturally occurring compound found in Aristolochia plants, is a well-established nephrotoxin and Group 1 carcinogen. Emerging evidence suggests a potential link between AA exposure and hepatocellular carcinoma (HCC), one of the leading causes of cancer-related mortality worldwide. This review critically evaluates current knowledge on AA's hepatic metabolism, its formation of persistent DNA adducts, and the induction of inflammatory responses in the liver. Based on preclinical and indirect human evidence, we propose a working hypothesis that AA may contribute to hepatocarcinogenesis through a dual mechanism: genotoxic (primarily via H-ras and p53 mutations resulting from AA-DNA adducts) and non-genotoxic (via chronic inflammation involving IL-6, TNF-α, and NF-κB activation, as well as epithelial-mesenchymal transition). We note, however, that these mechanisms remain to be validated in human cohorts and do not yet establish causality. Recent studies have identified novel mechanisms, including PDK4-mediated mitochondrial dysfunction, ferroptosis inhibition via p53 hijacking, and ARID1A deficiency as a susceptibility factor. A recent meta-analysis quantified a significantly increased risk of liver cancer following AA exposure in epidemiological studies. While direct causal evidence in humans remains limited, the high mutational burden observed in AA-exposed liver tissues warrants caution. Nevertheless, the primary public health priority pertains to the prevention of AA exposure. Further epidemiological and mechanistic studies are urgently needed.

Self-healing materials have attracted increasing attention as a strategy to enhance durability, extend service life, and reduce maintenance in advanced material systems. Among these, cellulose-based self-healing materials represent a sophisticated intersection between sustainable macromolecular chemistry and adaptive materials science. This review provides a synthesis of recent advancements in the field, systematically categorizing materials derived from cellulose raw materials. We evaluate the fundamental chemical strategies employed to achieve autonomous repair, distinguishing between extrinsic mechanisms-utilizing cellulose-based micro/nano-capsules to sequester healing agents-and intrinsic mechanisms governed by dynamic covalent chemistry (Schiff-base, boronic ester, Diels-Alder) and supramolecular interactions (hydrogen bonding, metal-ligand coordination, and host-guest assemblies). The analysis highlights how cellulose's hierarchical structure and abundant surface functionality are leveraged to overcome the traditional trade-off between mechanical toughness and healing efficiency. Particular emphasis is placed on the transition from simple structural hydrogels to sophisticated multifunctional systems. These include ultra-stretchable strain and pressure sensors for e-skin applications, biocompatible and injectable matrices for chronic wound management and stem cell delivery, and advanced anti-freezing eutectogels for performance in extreme environments. Furthermore, we explore the integration of cellulose into traditional sectors, such as self-healing concrete utilizing microbe-induced calcification and smart, eco-friendly coatings for corrosion protection. Finally, we discuss critical challenges, including environmental stability, scalability, and the development of standardized evaluation protocols, providing a roadmap for the next generation of bio-derived, sustainable and intelligent materials.

<i>Hippophae gyantsensis</i> Lian is an important native tree species in the "One River, Two Streams" valley of Tibet, valued for its ecological restoration potential and nutrient-rich fruits. However, this species has several limitations, including a long fruiting cycle (3-5 years to flowering and 10-15 years to reach peak fruit production), small fruit size, and numerous branch thorns. These traits hinder large-scale cultivation and mechanized harvesting, creating an urgent need for improved varieties with larger fruit and higher yield. In this study, we established an efficient <i>Agrobacterium</i>-mediated genetic transformation system for <i>H. gyantsensis</i> using hypocotyls as explants. Under optimized conditions (OD₆₀₀ = 0.5, AS = 200 μmol/L, infection time = 15 min), the transformation efficiency reached 36.67% (calculated as the number of PCR-positive plants divided by the total number of explants initially inoculated with <i>Agrobacterium</i>). A rooting rate of 12.5% was achieved using 100 mg/L rooting powder (ABT1) for 40 min, resulting in an overall success rate of approximately 4-5%. Furthermore, we identified and cloned two fruit-size-related genes, <i>Hgfw2.2</i> and <i>Hgfw3.2</i>, from <i>H. gyantsensis</i>. Heterologous expression of <i>Hgfw2.2</i> and <i>Hgfw3.2</i> in tomato decreased and increased fruit size, respectively, consistent with their regulatory roles in fruit development. Given the positive regulatory effect of <i>Hgfw3.2</i>, this gene was further transformed into <i>H. gyantsensis</i>. This study represents the first report of a stable genetic transformation platform for <i>H. gyantsensis</i>, providing a robust technical foundation for future molecular breeding and the development of improved, large-fruited varieties.

Neurological disorders, including Alzheimer's disease, Parkinson's disease, and stroke, remain major causes of global disability and mortality, with limited neuroprotective therapies available. Traditional Chinese medicine (TCM) offers multi-target therapeutic potential, but its mechanistic complexity requires systematic investigation using appropriate model systems. Zebrafish (<i>Danio rerio</i>) has emerged as a valuable vertebrate platform for TCM neuroprotection research due to its genetic homology with humans, optical transparency, and high-throughput screening compatibility. This review summarizes the application of zebrafish models in studying TCM for Alzheimer's disease, Parkinson's disease, cerebral ischemia, epilepsy, insomnia, depression, and spinal cord injury. Key findings indicate that TCM metabolites exert neuroprotective effects through multiple mechanisms, including anti-oxidative stress, anti-neuroinflammation, anti-apoptosis, neurotransmitter modulation, neurogenesis promotion, and vascular protection. Zebrafish models have proven particularly useful for high-throughput screening of active metabolites, real-time <i>in vivo</i> imaging of neurovascular processes, and rapid safety assessment. However, limitations such as the absence of a layered neocortex, differences in drug metabolism, and the predominantly acute nature of current models must be acknowledged. Addressing these challenges through model standardization, multi-omics integration, and cross-species validation will further enhance the translational relevance of zebrafish-based TCM research. This review provides a practical framework for leveraging zebrafish models to advance the mechanistic understanding and clinical development of neuroprotective TCM therapies.

RNA interference (RNAi) regulates gene expression through small RNAs that act via Argonaute-containing RNA-induced silencing complexes (RISCs). We previously found that short RNAs with G-rich 6mer seeds (e.g., GGGGGC and G5C) can kill cells by targeting C-rich 3' UTR seed matches in essential survival genes (SGs), a mechanism termed death induced by survival gene elimination (DISE). To assess therapeutic potential, we systemically delivered two DISE-inducing sRNAs, sG5C and sCAG (based on CAG trinucleotide repeats), using lipopolyplexes (LPPs) composed of low-molecular-weight polyethyleneimines and lipids. In mouse ovarian and prostate cancer models and a rat hepatocellular carcinoma model, LPP-delivered small RNAs (sRNAs) markedly reduced or eliminated tumors without harming normal tissues. Predicted SG targets were engaged in tumors. Transcriptomic analyses across 10 major human cancers showed that many sG5C-targeted SGs are consistently upregulated in tumors and increase with stage, revealing a therapeutic window. These results support LPP-delivered DISE-inducing sRNAs as a promising pan-cancer therapy.

MicroRNAs (miRNAs) are small, non-coding RNAs that regulate gene expression post-transcriptionally and play crucial roles in cancer biology and immune function. Among them, miR-150-5p has emerged as a key regulator with complex, context-dependent roles in both tumorigenesis and immune cell differentiation. This review provides a comprehensive synthesis of current knowledge on miR-150-5p, highlighting its dual function as a tumor suppressor or oncogene depending on cancer type and cellular context. We examine its involvement in hematologic malignancies and solid tumors, detailing the molecular mechanisms through which it influences proliferation, apoptosis, and metastasis. Particular emphasis is placed on the role of extracellular vesicle (EV)-associated miR-150-5p as a modulator of the tumor microenvironment (TME), including its impact on angiogenesis, immune evasion, and intercellular communication. We further explore miR-150-5p's regulation of key immune cell subsets-such as macrophages, dendritic cells, T cells, and natural killer cells-and its implications for anti-tumor immunity. Finally, we discuss the therapeutic potential and challenges of targeting miR-150-5p, including delivery barriers, off-target effects, and opportunities for personalized medicine. By integrating recent findings, this review underscores miR-150-5p's value as both a biomarker and a therapeutic target in cancer immunology.

<h4>Objectives</h4>Human enteroviruses (EVs) are associated with a broad spectrum of diseases, including hand, foot, and mouth disease. This study aimed to understand the epidemiology and genotypes/subgenotypes of EVs circulated in Dak Lak, Vietnam between 2022 and 2024.<h4>Methods</h4>Samples were collected from patients admitted to the inpatient and outpatient departments from 2022 to 2024 in Dak Lak of Vietnam. EVs were identified in all samples through real-time reverse-transcription polymerase chain reaction using a commercially available kit.<h4>Results</h4>EVs were detected in 96 samples in the period from 2022-2024 in total of 167 suspected samples; the EV-positive rate was 57.5%. The highest positivity rate for EVs was observed in children aged <5 years, reaching 96.8%, while the rates of EV infections showed no significant differences between genders. The monthly positivity rate for EV infection ranged from 0% to 84.6%, with a peak in September. The most frequently observed EVs were EV A71 (28.1%), Coxsackievirus A6 (28.1%), followed by Coxsackievirus A16 (16.7%) and Coxsackievirus A10 (10.4%), while the proportion of unidentified EV serotypes were 10.4%. The phylogenetic analysis of the VP1 gene of EV A71 strains revealed that two genotypes, B5 (93.7%) and C4 (6.3%), were found and all were closely related to variants causing epidemics of hand, foot, and mouth disease in southern Vietnam that circulated in 2023. Meanwhile, all CVA6, CVA16, and CVA10 strains belonged to D3, B1a, and C3 subgenotypes, respectively.<h4>Conclusion</h4>EV infection is highly prevalent among young children aged <5 years in Dak Lak. Further monitoring through techniques capable of subtyping all EVs is necessary to enhance the observation of these infections and their causes.

<title>Abstract</title> <p> <bold>Purpose</bold> Serum ferritin is an imperfect surrogate of hepatic iron burden, particularly in metabolic and inflammatory liver disease. We evaluated the diagnostic performance of serum ferritin against MRI-derived liver iron concentration (MRI-LIC) and characterized the biological correlates of biochemical–imaging discordance. <bold>Methods</bold> In this single-center retrospective cohort study, 695 adults evaluated for suspected iron overload between 2003 and 2024 underwent serum ferritin, transferrin saturation (TSAT), and HFE genotyping. Biochemical data were available for 664/695 patients, histopathology for 673/695, and paired ferritin–MRI measurements for 77/695. Diagnostic performance of ferritin was assessed by receiver operating characteristic (ROC) analysis and agreement metrics; independent predictors of MRI-defined iron overload (LIC ≥ 2.0 mg/g) were identified using Firth penalized logistic regression. <bold>Results</bold> Among 77 patients with paired data, ferritin showed moderate correlation with MRI-LIC (Spearman ρ = 0.66, p < 0.001) and good discrimination for iron overload (AUC 0.86, 95% CI 0.77–0.95). At 1,000 ng/mL, ferritin showed high specificity (88%) and negative predictive value (90%) but lower sensitivity (54%) and positive predictive value (47%); overall agreement was 82% (κ = 0.39). In multivariable Firth regression, TSAT—but not ferritin—retained association with MRI-defined iron overload. MRI-LIC varied nonlinearly across fibrosis stages, peaking in intermediate fibrosis (F2; Kruskal–Wallis p = 0.006). <bold>Conclusion</bold> Serum ferritin offers high negative predictive value but limited specificity and positive predictive value for hepatic iron overload. MRI-LIC provides direct tissue iron quantification and should be used when biochemical findings are discordant or dysmetabolic hyperferritinemia is suspected. </p>

Mounting evidence has associated gut microbial composition and systemic metabolic profiles with Parkinson's disease (PD), yet causal pathways and molecular mechanisms remain unclear. We employed a systematic two-stage analytical framework integrating multiple Mendelian randomization (MR) approaches, transcriptome and epigenome-wide regulatory analyses, and colocalization analysis to genetically dissect the microbiota-host regulatory network implicated in PD susceptibility. Initially, two-sample MR was performed using gut microbiota data from the MiBioGen consortium, followed by an independent replication MR analysis using mbQTL data from the Dutch Microbiome Project (DMP). The two-step MR approach was employed to identify potential mediation of blood metabolites. Summary data-based MR (SMR) methods were also used to pinpoint key blood metabolic genes and regulatory regions associated with PD risk, with the combination of PD GWAS data, blood expression quantitative trait loci (eQTLs), and DNA methylation quantitative trait loci (mQTLs). Colocalization analyses were performed to explore interactions between host metabolic gene expression and the gut microbiota using blood expression quantitative trait loci (eQTLs) and fecal microbial quantitative trait loci (mbQTLs). Our multi-stage analyses identified five microbial taxa with nominal positive associations with Parkinson's disease (PD), including class Clostridia, order Bacillales, and genera Clostridium sensu stricto 1, Dorea, and Lachnospiraceae UCG001. By contrast, three additional microbial genera (Butyricimonas, Defluviitaleaceae UCG011 and Marvinbryantia) exhibited nominal protective correlations with PD. Cross-cohort analyses further indicated a shared correlation pattern along the Clostridia-Clostridiales-Clostridiaceae/Lachnospiraceae taxonomics potentially linking to PD risk across both discovery and validation cohorts. Mediation analyses indicated that betaine may mediate the Lachnospiraceae UCG001-PD associations, while androstenediol monosulfate may serve as a mediator linking Clostridium sensu stricto 1 to PD. SMR analysis highlights genes like SH2B1, JUP, as key host metabolic genes with potential causal relevance to PD, with colocalization indicating genetic overlaps supporting their potential involvement in gene-microbiota interactions. These findings deepen our understanding of gut-brain axis dysregulation in PD and may provide novel insights for future mechanistic validation and targeted preventive or therapeutic investigations.

This study aims to identify pathogenic genes involved in diabetic nephropathy (DN) through multi-omics analysis of circadian rhythm disruptions to uncover targets for precision medicine. We conducted summary-data-based Mendelian Randomization (SMR) and colocalization analyses using multi-omics quantitative trait loci (QTL) data (methylation quantitative trait loci [mQTL] , expression quantitative trait loci [eQTL], protein quantitative trait loci [pQTL]) to explore genetic associations between circadian rhythm-related genes and DN. Genome-wide association study (GWAS) summary statistics were sourced from major consortia (discovery) and FinnGen (replication). Key gene expressions were validated in GEO (GSE96804). Transcription factors were predicted, and potential drugs (<i>via</i> Enrichr) were assessed through molecular docking. This study identified 395 methylation loci, 29 genes, and 5 proteins linked to disease using blood-derived QTL data. Replication confirmed 62 methylation loci, 5 genes, and 1 protein were associated with DN. Seven methylation loci influenced four DN-related genes (<i>BICC1</i>, <i>ANKIB1</i>, <i>KIF11</i>, <i>GNAI2</i>). Kidney tissues from DN patients showed elevated <i>GNAI2</i> and <i>KIF11</i> but reduced <i>PEBP1</i>. IRF2 was identified as a potential key transcription regulator. Drug predictions suggested calycosin, nitrofural, and nobiletin could target PEBP1, GNAI2, and KIF11, respectively. This SMR study suggested the causal role of circadian rhythm dysfunction in DN. Future research should focus on identifying specific genetic drivers, and exploring therapeutic strategies targeting circadian pathways to mitigate DN progression.

Chronic kidney disease (CKD) encompasses multiple pathogenic mechanisms manifested by inflammation, fibrosis, oxidative stress and cell death. We previously showed that circulating protein CD5L (or AIM) ameliorates acute kidney injury, so we explored its effect in a mouse model of unilateral ureteral obstruction (UUO)-induced renal fibrosis. Here, we show a unique kidney-protective pathway mediated by CD5L. CD5L is endocytosed into renal epithelial cells, where it reduces oxidative stress, decreasing cell injury and death. This effect is supported by both a cysteine-dependent direct antioxidant activity of CD5L and enhancement of Nrf2-associated antioxidant responses. In addition, our data suggest that suppression of sphingomyelinase activity and reduction of cellular ceramide may contribute, at least in part, to CD5L-associated augmentation of Nrf2 nuclear transport. These effects depend on the reactive cysteine residue on the CD5L surface. Consistent with these findings, recombinant CD5L treatment in UUO mice reduces sphingomyelinase activity, activates Nrf2, and lowers oxidative stress, alleviating inflammation, fibrosis, and kidney injury. Our findings uncover a novel antioxidant pathway mediated by CD5L with potential implications for CKD-associated fibrotic mechanisms.

Ferroptosis, a unique form of regulated cell death driven by iron-dependent lipid peroxidation, and macrophage polarization, which reflects the high plasticity of the immune system, represent two prominent frontiers in contemporary life sciences. Emerging evidence suggests a profound bidirectional interplay between these processes, mediated by iron metabolism reprogramming, lipid signaling molecules, and complex molecular axes such as RAGE-STAT3. This review systematically summarizes the biological mechanisms of ferroptosis and macrophage polarization, delving into their interaction within the tumor microenvironment, inflammatory responses, and cardiovascular and neurodegenerative diseases (e.g., Alzheimer's and Parkinson's). We highlight how M1 macrophages induce ferroptosis through pro-inflammatory cytokines and reactive oxygen species, while M2 macrophages inhibit it by modulating iron homeostasis and antioxidant capacity. Finally, we discuss therapeutic strategies targeting the ferroptosis-macrophage polarization axis, providing a theoretical foundation and novel perspectives for developing precise medical interventions.

The emergence of resistance against first-generation non-nucleoside reverse transcriptase inhibitors (NNRTIs) necessitates the development of novel scaffolds with improved antiviral efficacy. In this study, a series of pyridinone derivatives were designed and synthesized as potential NNRTIs. The antiviral evaluation against wild-type HIV-1 in MT-4 cells revealed that most of the compounds displayed limited or no activity, often accompanied by cytotoxic effects. However, two derivatives, 4e and 4i, demonstrated remarkable inhibition of HIV-1 replication, with 4i exhibiting low-nanomolar potency against HIV-1 (EC₅₀ = 5.3 nM), having almost one half of the Rilpivirine activity. Molecular docking confirmed critical hydrogen bonding interactions with Lys103 and π-π stacking with Trp229 and Tyr181, while 300 ns molecular dynamics simulations verified the stability of the compound-enzyme complexes. Collectively, these findings highlight the pyridinone scaffold, particularly compound 4i, as a promising lead for further optimization toward next-generation NNRTIs.

<h4>Purpose</h4>Anti-vascular endothelial growth factor (anti-VEGF) therapy is an important treatment for type 1 retinopathy of prematurity (T1ROP). We previously found that regulation of VEGF signaling increased developmental vascularization into the temporal peripheral retina while reducing intravitreal neovascularization in T1ROP. To address our hypothesis that extension of retinal vascularization by early treatment with anti-VEGF could reduce T1ROP, we determined the effects of anti-VEGF therapy on retinal vascular extension in both retinal hemispheres.<h4>Design</h4>Retrospective, masked, nonrandomized, matched, comparative case series.<h4>Subjects</h4>Over 3 years, 34 of 382 screened infants developed bilateral T1ROP. Consent was obtained from parents of 21 infants for infants receiving bevacizumab 0.25 mg OU. Seven treated eyes (7 infants) had suitable imaging and were included in the treatment group. Seven untreated eyes (7 infants) without T1ROP were randomly selected after matching for birthweight and postmenstrual age from the 382 screened infants and assigned to the control group.<h4>Methods</h4>Infants had gradable 100° field-of-view fundus photographs of both retinal hemispheres at 2 time points: baseline before treatment, if administered, and follow-up, 12 to 35 days later. No eyes with vascularization to the ora at baseline or follow-up were included. Retinal vascular extent was measured in pixels from the optic disc margin to the temporal or nasal vascular borders and divided by the optic disc diameter in pixels to create disc diameter units (DD). Measurements were confirmed by a second masked analyst. The Mann-Whitney U test and Wilcoxon signed-rank test were used for statistical analysis.<h4>Main outcome measures</h4>Temporal and nasal vascular extension in DD.<h4>Results</h4>Relative to control eyes, treated eyes had greater nasal (1.27 ± 0.59 vs 0.55 ± 0.44 DD; P = .038) and temporal (1.69 ± 1.16 vs 0.62 ± 0.49 DD; P = .04) extensions. There was greater disparity in vascular extension between nasal and temporal hemispheres in the treated group than in the control group (0.98 ± 0.64 vs 0.10 ± 0.10 DD; P = .005). Nasal extension was greater than temporal extension in 3 of 7 treated vs 1 of 7 control eyes.<h4>Conclusions</h4>Treated eyes had significantly greater vascular extension in both hemispheres than control eyes, with greater nasal-temporal asymmetry. Although limited by sample size, our findings raise the question of uneven anti-VEGF diffusion in the infant vitreous. Further studies testing injection site, needle length, anti-VEGF agent, and molecular size differences and further follow-up periods are indicated.

Antiangiogenic therapy remains a challenging issue in the treatment of glioblastoma (GBM). Effective therapies are in urgent need to improve prognosis of GBM patients. The high heterogeneity of GBM is both the cause of its unavoidable therapeutic resistance and the result of extensive genomic dysregulation, of which abnormal transcription factor (TF) networks are recognized to be the culprit. Herein, based on the heterogeneity of GBM, we identified the key TF serum response factor (SRF), which is closely associated with the formation of the GBM vascular microenvironment, from a large-scale dataset. Mechanistically, the 133-240aa region of SRF binds to the transcriptional cofactor P54nrb and undergoes phase separation to form a transcription complex, which upregulates OLFML3 by binding to its enhancer and promoter regions. The secreted extracellular matrix (ECM) glycoprotein OLFML3 activates endothelial cells (ECs) by degrading and remodeling the ECM, releasing proangiogenic factors, promoting intercellular adhesion, and directly acting on ECs to promote angiogenesis. The hyperplasia vasculature, in turn, promotes the infiltration of M2-polarized macrophages, leading to the formation of an immunosuppressive microenvironment. This study comprehensively elucidates the pivotal role of SRF in GBM angiogenesis. Targeting the SRF/P54nrb/OLFML3 axis holds promise for developing novel antiangiogenic strategies to improve GBM treatment outcomes.

Gastric cancer (GC) remains a formidable global health challenge, characterized by pronounced molecular heterogeneity, late-stage diagnosis, and limited durable responses to existing therapies. This review synthesizes recent advances in GC research through an integrated, multidisciplinary lens, spanning tumor biology, microenvironmental dynamics, and therapeutic innovation. We first consolidate updated histopathological and molecular classification systems, highlighting oncogenic programs that underpin GC development, including Hippo-YAP signaling and emerging neural-stem cell interactions. We then examine the immunosuppressive tumor microenvironment, emphasizing the dynamic crosstalk among tumor-associated macrophages, regulatory T cells, tertiary lymphoid structures, and cancer-associated fibroblasts that collectively drive metastatic dissemination and therapeutic resistance. Emerging biomarker-guided strategies, including CLDN18.2-targeted therapies, dual immune checkpoint blockade, and engineered cellular therapies, are critically discussed alongside rational combination approaches designed to overcome resistance. Beyond canonical paradigms, we highlight transformative frontiers, such as cancer neuroscience, microbiome-driven immune modulation, and spatially resolved multiomics technologies, that enable high-resolution mapping of cellular interactions. Finally, we critically assess translational barriers, including organ-specific metastatic tropism and resistance evolution, and propose that the convergence of deep molecular profiling, neural-immune modulation, and AI-enabled computational oncology will be central to advancing precision medicine for GC. This integrated framework aims to accelerate the development of mechanism-based combination therapies.

Prostate cancer (PCa) is frequently accompanied by benign prostatic hyperplasia (BPH), highlighting the need to reassess their correlation in tumor risk, malignant progression, and immune status to improve the early diagnosis and treatment of PCa. In this study, single-cell RNA sequencing and spatial transcriptomics were used to analyze the potential association between normal, BPH, and PCa tissues. Our study revealed a continuous transformation map of luminal epithelial cells from hyperplasia to malignancy in BPH and PCa, accompanied by a persistently suppressive immune microenvironment. In the lesion nodules, T cells showed a high degree of infiltration yet showed activation retardation and functional exhaustion. Macrophages were also significantly infiltrated, exhibiting significant M2 polarization characteristics, and inflammatory signaling pathways related to immune escape were activated. Natural killer (NK) cells and B cells were partially activated, yet the low abundance of NK cells and B cells resulted in functional limitations. Our results revealed the dynamic changes of the immune landscape during the occurrence and progression of PCa. Our classification and prognostic models provide a theoretical basis for immunomodulatory and personalized therapies and provide new tools for the early diagnosis and intervention of PCa.

<b>Major</b> depressive disorder (MDD) and anxiety disorders are increasingly understood as conditions involving complex metabolic dysregulation across multiple biological domains. This review aimed to synthesize current clinical and translational evidence on amino acid metabolism, lipid metabolism and short-chain fatty acids (SCFAs) as potential biomarkers, and components of integrative metabolic profiling in these disorders. A structured narrative approach was applied, focusing on studies assessing metabolomic alterations, their clinical correlates and their potential role in patient stratification, and treatment response. The available evidence indicates that amino acid disturbances, particularly within the tryptophan-kynurenine pathway, represent the most consistent and clinically interpretable findings. Lipid-related alterations, especially involving long-chain polyunsaturated fatty acids, provide complementary insights into membrane function, inflammation and neuroplasticity. In contrast, SCFAs appear to function as context-dependent markers rather than robust standalone biomarkers, with their clinical relevance depending on biological matrix, metabolic context and host-microbiota interactions. Importantly, most studies assess individual metabolites rather than integrated metabolic profiles, limiting their interpretability within a metabolomic framework. Overall, current evidence supports a shift toward integrative biomarker models that combine metabolic data with selected molecular and clinical parameters. Future research should focus on standardized, reproducible profiling approaches to enable biologically informed stratification and personalized treatment strategies.

No abstract available.

Hemochromatosis, a disorder caused by excessive iron absorption, was first described in the mid-1800s as bronze diabetes and pigmentary cirrhosis. The term hemochromatosis was believed to have been coined by von Recklinghausen in 1889. A genetic origin of this metabolic problem was first suggested in the 1930s. In 1977, Simon et al reported the association between an HLA class I–like molecule and the presumed hemochromatosis gene on chromosome 6p, establishing the genetic basis of what is now referred to as hereditary hemochromatosis. Although one of the most common genetic disorders in the United States, affecting over 1 million people, hereditary hemochromatosis is often an incidental finding during routine laboratory iron measurements or the diagnostic workup of other conditions; however, increasing awareness of hereditary hemochromatosis has also contributed to early detection. Early diagnosis allows for intervention before tissue damage occurs due to excessive iron deposition. Iron can deposit in the liver, pancreas, heart, joints, and other endocrine organs if left untreated. Hereditary hemochromatosis is currently classified into 4 major types, encompassing 5 distinct molecular subtypes. These classifications are based on age of onset, underlying genetic mutation, and mode of inheritance. Type I hereditary hemochromatosis is considered the classic form of the disorder, and the onset of symptoms begins in adulthood. Loss-of-function mutations in the hereditary Fe [iron] (HFE) gene are present in approximately 70% of patients diagnosed with hereditary hemochromatosis. This form of hereditary hemochromatosis disproportionately affects males, although females can also be affected. Type 2 hereditary hemochromatosis is frequently called juvenile hemochromatosis, as symptoms begin in childhood, and the disease is typically more clinically severe. Unlike type I hereditary hemochromatosis, this form shows no sex preference. Type 2 hereditary hemochromatosis has 2 subtypes: 2a and 2b. Type 2a hereditary hemochromatosis is caused by mutations in the gene initially known as hemojuvelin (HJV) and is now referred to as HFE2. Type 2b hereditary hemochromatosis is due to hepcidin antimicrobial peptide (HAMP) gene mutations. Type 3 hereditary hemochromatosis typically has an onset at approximately 30 years of age and is caused by mutations in the transferrin receptor 2 (TFR2) gene. Type 4 hereditary hemochromatosis, also known as ferroportin disease, is caused by mutations in the ferroportin/solute carrier family 40 member 1 (SLC40A1) gene. Ferroportin is an iron transmembrane transport protein, and Type 4 hereditary hemochromatosis is the only known form of hemochromatosis that can be inherited in an autosomal dominant fashion. The onset of type 4 hereditary hemochromatosis typically occurs in mid-adulthood.

Genetic risk factors for Parkinson's disease (PD) remain under-characterized in East Asian populations. We assembled a Taiwanese case-control cohort (2245 PD; 2147 controls), genotyped participants using the Illumina NeuroBooster Array, and imputed 7.6 million variants with the Taiwan Biobank reference panel. Logistic-regression GWAS identified genome-wide significant associations at SNCA and MCCC1, with the lead SNCA signal located in intron 4; conditional and joint analyses suggested an additional 5' SNCA component. We observed suggestive associations at GCH1, PPARGC1A, and GALNT13. Locus-focused haplotype analyses refined the SNCA signal, delineating an East Asian-enriched risk haplotype, and confirmed risk effects of LRRK2 p.G2385R and p.R1628P, including evidence consistent with a gene-dosage effect. A European-derived PRS showed modest discrimination in our cohort (AUC 0.59), while incorporating East Asian and Taiwan-relevant variants improved performance (AUC 0.62). Together, these results define the genetic architecture of PD in Taiwan, highlight shared and population-enriched risk components, and support ancestry-aware PRS construction for improved risk stratification.

<h4>Objective</h4>The authors evaluated changes associated with a 3-week interdisciplinary intensive outpatient program that delivers integrated neuropsychiatric rehabilitation for veterans and first responders with persistent postconcussive and co-occurring symptoms after remote traumatic brain injury.<h4>Methods</h4>In this retrospective cohort study, 128 patients completed patient-reported measures, including the Neurobehavioral Symptom Inventory (NSI), PTSD Checklist for DSM-5 (PCL-5), and Life Satisfaction Questionnaire-11 (LiSat-11), and objective assessments, comprising Addenbrooke's Cognitive Examination-III (ACE-III), gait speed, computerized dynamic posturography (for conducting the Sensory Organization Test [SOT]), and Modified Clinical Test of Sensory Interaction in Balance (mCTSIB). Paired t tests were used to compare intake with discharge measures, and a linear mixed-effects model was used to examine NSI scores over time with covariate adjustment.<h4>Results</h4>The NSI total score decreased by 15.6 points (41%, Cohen's d=1.11); ACE-III scores improved by 6.8 points (Cohen's d=1.41); and forward and backward gait speeds, SOT composite scores, and mCTSIB condition 4 scores showed significant improvements (Cohen's d=0.68-0.87). The proportion of patients screening at or above a common PTSD threshold on the PCL-5 declined from 51% at intake to 20% at discharge, and higher baseline PCL-5 scores were associated with higher NSI scores across time points (p<0.001).<h4>Conclusions</h4>These findings are consistent with multidomain benefits of intensive, interdisciplinary neuropsychiatric rehabilitation in this population. These results support the potential of integrated care models to reduce symptom burden and improve cognitive, mobility, vestibular, and life satisfaction outcomes over 3 weeks, while underscoring the close linkage between posttraumatic stress severity and neurobehavioral symptom burden.

N6‑methyladenosine (m⁶A) modifications are key epigenetic regulatory mechanisms in mammals and serve key roles in both normal skeletal development and the pathogenesis of skeletal disorder. The dynamic and reversible regulation of m⁶A relies on three core factors: Methyltransferases (writers), demethylases (erasers) and m⁶A‑binding proteins (readers), which collectively ensure proper physiological functions. Despite this, the functions and regulatory mechanisms of numerous m⁶A‑associated factors in skeletal diseases remain insufficiently understood. m6A modification maintains bone homeostasis during skeletal development primarily by regulating the balance between osteoblasts and osteoclasts. Under pathological conditions, dysregulated m⁶A modification contributes to aberrant osteoclast proliferation and chondrocyte apoptosis, leading to bone loss and cartilage degeneration. These pathological changes are key contributors to common types of skeletal disorder, including osteoporosis, osteoarthritis, rheumatoid arthritis and intervertebral disc degeneration, imposing a burden on human health. Non‑coding RNAs are major targets of m⁶A modification and their interactions exert post‑transcriptional regulation in skeletal biology. The present review summarizes the roles and mechanisms of m⁶A modification in skeletal diseases and highlights its therapeutic potential, offering novel perspectives for disease prevention and treatment.

In p53-deficient cancers, targeting cholesterol metabolism has emerged as a promising therapeutic approach, given that p53 loss dysregulates sterol regulatory element-binding protein 2 pathways, thereby enhancing cholesterol biosynthesis. While cholesterol synthesis inhibitors such as statins have shown initial success, their efficacy is often compromised by the development of acquired resistance. Consequently, strategies are being explored to disrupt cholesterol homeostasis more comprehensively by inhibiting its synthesis and intracellular transport. In this study, we investigate a previously underexplored function of PI5P4Ks, which catalyzes the conversion of PI(5)P to PI(4,5)P₂ at intracellular membranes. Our findings reveal that PI5P4Ks play a key role in facilitating lysosomal cholesterol transport, regulating lysosome positioning, and sustaining growth signaling via the mechanistic target of rapamycin (mTOR) pathway. While PI5P4Ks have previously been implicated in mTOR signaling and tumor proliferation in p53-deficient contexts, this work elucidates an upstream mechanism that unifies these earlier observations.

Chronic hepatitis B (CHB), a major global public health burden, is characterized by the presence of high circulating levels of non-infectious subviral particles (SVPs). These SVPs are mainly composed of small hepatitis B surface antigen (SHBs) and are associated with a dysfunctional HBV-specific immune response. Previous studies exploring the cellular mechanisms and molecular pathways by which SVPs are secreted from hepatocytes have utilized either non-hepatic cells or whole-genome HBV-integrated hepatoma cell lines in which SHBs can also be secreted as a component of the virion envelope. In this study, we provide a detailed examination of SHBs trafficking using biochemical approaches coupled with high-resolution confocal microscopy and live-cell imaging via a novel fluorophore-tagged SHBs probe in both HepG2 and primary human hepatocytes stably expressing SHBs. Our results provide evidence that SHBs utilize a modified ER-Golgi secretory pathway with transit from the ER to the multivesicular body and lysosomal/late endosomal compartments. Importantly, we identified the autophagy adapters ATG5 and LC3B, and the ER-phagy protein atlastin 3 (ATL3), central to mediating SHBs secretion. These findings suggest that SHBs is trafficked through a non-canonical path for secretion and provides new potential therapeutic targets for the treatment of CHB.

Entacapone is a catechol-O-methyltransferase (COMT) inhibitor, widely used for the symptomatic treatment of Parkinson's disease. However, it presents setbacks associated with a short half-life and a poor blood-brain barrier permeability. In order to surpass these limitations, entacapone was encapsulated in lipid-polymer hybrid nanoparticles (LPHNPs) functionalized with vitamin E (TPGS), using different liquid lipids as olive oil and coconut oil, and the co-surfactant and penetration enhancer Transcutol® HP. Depending on the system chosen, the nanoformulations presented differences in the hydrodynamic sizes (D_DLS) after synthesis, and an increasing particle size in the following order: Transcutol® HP< olive oil < coconut oil. The nanoformulations presented interaction with mucins and serum proteins, which anticipates a good permeability for oral and intranasal administration. No cytotoxic effects were observed in human neuroblastoma (SH-SY5Y), human hepatocellular carcinoma (HepG2), or squamous cell carcinoma (RPMI 2650) cell lines for concentrations below 10 µM. Furthermore, olive oil and Transcutol® HP nanoformulations maintained the COMT inhibition effect in HepG2 cells and presented antioxidant and iron chelation properties in SH-SY5Y cells. An increase in entacapone permeability was observed for Transcutol® HP nanoformulations using in vitro RPMI cells. In vivo study using Caenorhabditis elegans as a model demonstrated survival percentages > 80% after acute exposure to the nanoformulations in concentrations up to 40 μM in both wild-type (N2) and parkinson's disease model (WLZ3) strain, and antioxidant activity for HTT@Ent (40 μM) formulation in wlz3 strain. Overall, this work shows that the encapsulation of entacapone could be an interesting alternative in solving the drug performance by improving its physicochemical properties and permeability.

Pediatric acute myeloid leukemia (AML) is a heterogeneous hematologic malignancy that accounts for about 15%-20% of childhood leukemias. Despite therapeutic advances, relapses remain common, and survival for high-risk patients is below 60%. Unlike adult AML, pediatric AML displays distinct genetic mutations, including FLT3-ITD, NPM1, KMT2A rearrangements, and core-binding factors (CBF) fusions, as well as extensive epigenetic dysregulation. Aberrant DNA methylation, histone modifications, and altered non-coding RNA expressions disrupt hematopoietic differentiation and activate oncogenic transcriptional networks. Recent advances in silico transcriptomic analysis have transformed the study of pediatric AML by integrating gene expression and epigenetic data to identify molecular drivers and regulatory networks. Computational RNA-seq pipelines and pathway analyses have highlighted key epigenetic regulators, including DNMT3A, TET2, and HDACs, as potential therapeutic targets. Multi-omics approaches combining transcriptomic, methylomic, and chromatin accessibility data are increasingly used to define biomarkers for diagnosis, prognosis, and therapeutic response. This review provides a comprehensive overview of the molecular and epigenetic landscape of pediatric AML, emphasizing the power of in silico transcriptome analysis to uncover disease mechanisms, refine patient stratification, and guide the development of precision-based epigenetic therapies aimed at improving long-term outcomes in children with AML.

γδT cells comprise 1-10% of circulating T cells, are further enriched in barrier tissues such as the gut and skin, and also form a critical component of lymphoid stress surveillance. γδ T cells have been gaining prominence as an alternative to αβ T cells for cancer immunotherapy. Such popularity may be attributed to multiple factors including the MHC-independent nature of γδ T activation, which differentiates them from αβ T cells and underlies their lack of alloreactivity. In this Review, we describe efforts to optimize the potential of γδ T cells as a cancer immunotherapeutic. We discuss the impact of disease burden on efficacy and the contexts in which unmodified γδ T cells have succeeded or failed to yield durable responses. Finally, we explore options for enhancing γδ T cell efficacy including combinations with other treatments and engineering strategies to capitalize on their potency.

Parafoil systems are increasingly deployed in military and civilian missions that require high-precision trajectory tracking. However, conventional controllers based on linear strategies and limited decision information often fail to cope with task complexity, and existing reinforcement learning studies rarely consider the systematic construction of training task sets, thereby limiting generalization. To address these issues, this paper proposes a Benchmark Task Set Generation (BTSG) algorithm that builds high-quality task sets with diverse and balanced complexity for robust controller training and comprehensive evaluation, formulates the trajectory tracking problem as a Markov decision process with a 46-dimensional observation space and a dense reward structure to enrich decision-support information and improve learning efficiency, and develops a BTSG-SAC framework that integrates BTSG with Soft Actor-Critic (SAC) for controller training. Simulation results show that BTSG-SAC markedly outperforms PID and PPO controllers on the generated benchmark task set, reducing distance deviation by 50.7% and 33.2% under noise-free conditions; under sensor noise, BTSG-SAC attains a 98% success rate, whereas PPO reaches 77% and PID fails to achieve successful tracking. Overall, this paper extends the application of reinforcement learning to parafoil control and offers generalizable methodologies for future autonomous control research.

Post-hepatectomy liver failure (PHLF) remains a serious complication following liver resection, yet early prediction is an unmet clinical need. This prospective study enrolled 151 patients at elevated risk of PHLF (major hepatectomy, thrombocytopenia, or hyperbilirubinemia) and evaluated serum antithrombin III (ATIII) activity, measured preoperatively and on postoperative days (PODs) 1, 2, 3, and 5, as an early predictive marker. PHLF, diagnosed according to the International Study Group of Liver Surgery criteria, occurred in 35 patients (23.2%). ATIII activity was evaluated as raw values and as the percentage change from the preoperative baseline. Raw ATIII activity was significantly lower in the PHLF group at all time points (P < 0.001). The POD 3 ATIII decrease was significantly greater in the PHLF group (36% vs. 29%, P = 0.041). Multivariable logistic regression identified ATIII change from baseline ≥ 30% at POD 3 (odds ratio 3.04, P = 0.021), ALBI grade B (OR 2.77, P = 0.031), and ICG R-15 ≥ 15% (OR 3.50, P = 0.034) as independent risk factors for PHLF. The multivariable model demonstrated acceptable discriminative performance (apparent AUC 0.750; bootstrap-corrected AUC 0.730). These findings suggest that early postoperative decline in ATIII activity could serve as an early biomarker for identifying patients at increased risk of PHLF, potentially enabling timely intervention.

SWI/SNF-deficient cancers characterized by dual SMARCA4/SMARCA2 loss, including subsets of lung adenocarcinoma and small cell carcinoma of the ovary, hypercalcemic type, represent a major therapeutic challenge. While SMARCA2 paralog-targeting strategies exist for single-subunit loss of SMARCA4, dual SMARCA4/SMARCA2-deficient tumors lack actionable targets. Here, we identify the CHD3/NuRD nucleosome remodeling complex as a critical synthetic lethal vulnerability in these malignancies. Through integrated genomic analyses, we demonstrate that in the absence of SWI/SNF, CHD3 acts as an essential "epigenetic brake" specifically at the enhancer of the cell polarity regulator PARD3B. Loss of CHD3 triggers aberrant chromatin hyper-accessibility and toxic derepression of PARD3B. We further elucidate that PARD3B accumulation is a critical mediator of cell death, which is strongly associated with the attenuation of MYC signaling signatures. This attenuation likely arises from the spatial perturbation of upstream signaling hubs or secondary cellular responses, reducing MYC transcriptional output without degrading the protein itself. Therapeutically, CHD3 depletion led to robust tumor regression in dual SMARCA4/SMARCA2-deficient xenografts, validating the in vivo mechanism of PARD3B upregulation. Collectively, our study defines a novel mode of synthetic lethality driven by "gain-of-toxicity" rather than the loss of survival signals, uncovering a fatal cross-complex dependency. We propose that targeting CHD3 to trigger toxic PARD3B derepression offers a promising therapeutic avenue for treatment-refractory dual SMARCA4/SMARCA2-deficient cancers.

<h4>Objectives</h4>To assess the potential role of glutamine metabolism-related genes (GMRGs) in the progression, prognosis, and immune microenvironment of clear cell renal cell carcinoma (ccRCC).<h4>Methods</h4>Publicly available single-cell RNA sequencing dataset was obtained, and differential gene expression analysis across tumors were performed. A cross scale disease driver gene validation chain and spatial transcriptome analysis with glutamine metabolism was further analyzed. 17 gene sets (M10295) correlated with glutamine metabolism were derived. The prognostic model of ccRCC constructed according to the differentially expressed GMRGs (DEGMRGs). GO and KEGG data were applied and the "GSVA" R package was used to analyze the characteristics of immune cells infiltration. In vitro validation study was performed in various RCC cell lines. Two shRNA plasmids were independently transfected into cells and examined by Transwell migration assay, proliferation assays and colony information tests. Glutamine levels and its related metabolism genes were also detected.<h4>Results</h4>ALDH18A1 may serve as a tumor-specific glutamine metabolic marker in epithelial-derived malignancies, and spatial transcriptome analysis reveals enhanced glutamine metabolic activity in the tumor boundary region. The DEGMRGs-based prognostic model tailored for ccRCC delineated a stark contrast in the prognosis of high-risk group and low-risk group. Patients exhibiting high ALDH18A1 TPM demonstrating poorer prognosis compared to those with low levels. The in vitro study validated the high expression of ALDH18A1 in RCC cell lines. ALDH18A1 could significantly inhibit the proliferation and migration of RCC cells, suppress the glutamine metabolism in RCC cells. Additionally, subcutaneous tumor experiments in mice further confirmed the oncogenic-promoting effects of ALDH18A1 on RCC with the crosstalk of M2 macrophages.<h4>Conclusions</h4>ALDH18A1 could serve as a crucial role in precise classification, treatment response, and prognosis of patients with ccRCC by suppressing the proliferation and glutamine metabolisms in tumor cells and constructing tumor microenvironment.

<h4>Background</h4>The etiology of gestational diabetes mellitus (GDM) is constituted by both type 2 diabetes (T2D)-dependent and T2D-independent mechanisms. However, existing studies have evaluated the impact of T2D-dependent loci only for GDM, which limits the power to assess to what extent genetic variants or biological pathways are specific to GDM.<h4>Methods</h4>We subtracted the genetic effects of a genome-wide association study (GWAS) data for T2D from a GWAS data for GDM to reveal loci linked with T2D-independent components of GDM using genomic structural equation model (SEM). In the discovery stage of GWAS-by-subtraction, we used GWAS summary statistics of GDM and T2D from the FinnGen study as input latent variables (16,802 GDM cases and 237,816 controls; 71,728 T2D cases and 369,007 controls). In the replication stage, we adopted the summary statistics from a previously reported GWAS for GDM and T2D as input (21,263 GDM cases and 301,918 controls; 50,409 T2D cases and 523,897 controls). We functionally annotated variants with genome-wide significance, and performed comprehensive analyses including transcriptome-wide and proteome-wide associations, summary-data-based mendelian randomization, linkage disequilibrium score regression, and Mendelian randomization, to explore genetic patterns for T2D-dependent and -independent components of GDM.<h4>Results</h4>We found 69 independent genome-wide significant loci associated with the T2D-independent components of GDM for discovery stage, of which 49 SNPs are not significant in the GWAS for GDM used as input. T2D-independent components of GDM showed only genetic correlation with birth weight dependent on maternal genetic effect (rg = 0.19; P = 0.01), whilst T2D-dependent components of GDM showed genetic correlation with birth weight dependent on fetal genetic effect (rg = -0.22; P = 4.76 × 10^- 7). Loci of T2D-independent components of GDM effects map to genes related to islets of Langerhans, neural stem cells, blood levels of mannose, abundance of Streptococcus thermophilus and Bacteroides vulgatus, glucose and carbohydrate homeostasis, and the MAPK cascade and adenylate cyclase-activating G protein-coupled receptor signaling pathway.<h4>Conclusions</h4>We identified independent genetic components, loci and genes of GDM that may have been previously masked by T2D, providing more accurate targets for early detection and management of GDM.

<h4>Purpose</h4>A history of coronary heart disease (CHD) increases the risk of Brain-Heart Syndrome (BHS) after acute stroke, partly through heightened inflammatory responses. Evolocumab, a PCSK9 inhibitor, has anti-inflammatory properties, but its transcriptomic effects in BHS patients with CHD remain unclear. This study aims to identify evolocumab-associated transcriptomic changes and inflammation-related biomarkers in this population.<h4>Patients and methods</h4>Blood samples from 24 BHS patients with CHD history (12 receiving rosuvastatin alone, 12 receiving rosuvastatin plus evolocumab) underwent transcriptomic sequencing. Candidate biomarkers were identified via differential expression and machine learning, with functional enrichment and immune infiltration analyses conducted.<h4>Results</h4>Four candidate biomarkers were identified: <i>WHRN (DFNB31)</i>, <i>IL12A</i>, and <i>ASB14</i> were upregulated, while <i>TMED7-TICAM2</i> was downregulated in the evolocumab combination group. These genes were enriched in pathways related to cell metabolism, signal transduction, and immune regulation. Immune infiltration analysis showed modest but detectable changes in B-cell subsets. External validation confirmed differential expression of these candidate biomarkers in CAD patients.<h4>Conclusion</h4>This pilot study provides preliminary insights into the molecular mechanisms of evolocumab in treating Brain-Heart Syndrome with a coronary heart disease history, identifying four inflammation-related biomarkers. These findings suggest potential targets for future investigation; however, given the exploratory nature and small sample size, further experimental and clinical validation is required before any therapeutic application.

Generalized pustular psoriasis (GPP) is a severe and potentially life-threatening form of psoriasis. Although spesolimab, an inhibitor of the interleukin (IL)-36 pathway, has been approved for the treatment of GPP, access to this agent remains limited. Vunakizumab, a humanized IgG1/κ monoclonal antibody that selectively neutralizes interleukin (IL)-17A inhibitors, is not yet approved for GPP. We herein report a 72-year-old male with a 40-year history of plaque psoriasis who developed GPP refractory to methotrexate combined with guselkumab and acitretin (20 mg/day). Following switching to vunakizumab (240 mg intravenously every 2 weeks) in combination with acitretin (40 mg/day), the patient achieved a GPPASI 75 response within 2 weeks and near-complete clearance (GPPASI ≈ 100) by week 12. Six induction doses of vunakizumab were administered, and acitretin was tapered to 20 mg/day for long-term maintenance. Throughout 36 weeks of follow-up, no disease relapse or drug-related adverse events were observed. This case, together with a review of the literature, provides support for IL-17A blockade combined with acitretin as a feasible and fast-acting regimen for refractory GPP in elderly patients.

<h4>Background</h4>The HAUS family proteins (HAUS1-HAUS8) are essential for mitotic spindle microtubule nucleation. Although HAUS dysregulation has been linked to tumor progression, whether these oncogenic functions are conserved or tissue-specific remains unclear. Therefore, a pan-cancer analysis is needed to identify universal HAUS drivers and context-dependent members for precision oncology.<h4>Methods</h4>We systematically evaluated the expression and prognostic significance of all eight HAUS genes across 33 cancer types using TCGA data. Focusing on liver hepatocellular carcinoma (LIHC), we identified molecular subtypes based on HAUS co-expression patterns and characterized their associations with clinicopathological features, the tumor microenvironment (TME), immune checkpoint expression, and therapeutic response. A HAUS-related prognostic signature was developed using LASSO and Cox regression analyses and validated in independent ICGC cohorts. Regulatory relationships among HAUS members and key signature genes were experimentally validated using immunohistochemistry and Western blotting.<h4>Results</h4>Most HAUS members were overexpressed across multiple cancers and correlated with poor clinical outcomes. In LIHC, two HAUS-based subtypes differed significantly in survival, clinicopathological profiles, immune features, mutational burden, stemness indices, and predicted therapeutic response. A prognostic signature comprising DTYMK and SPP1 effectively stratified LIHC patients into distinct risk groups, with the high-risk group showing significantly worse survival. A nomogram integrating the HAUS-related risk score with clinicopathological variables demonstrated strong predictive performance. Recombinant osteopontin induced HAUS1 and DTYMK expression in LIHC cell lines, supporting a functional SPP1-HAUS1/DTYMK axis.<h4>Conclusion</h4>This study establishes the broad oncogenic relevance of HAUS genes across cancers and demonstrates that a HAUS-based signature enables prognostic stratification in LIHC.

Digital droplet-based bioassays (ddAssays) have been applied in numerous fields, though many existing microfluidic platforms are tailored to specific assay classes and require complex workflows involving multiple processing steps. Here, we present a fully integrated microfluidic platform for generic digital bioassays that combines droplet generation and storage for on-chip incubation and optical readout on a single disposable device. This microfluidic platform enables the parallel execution of test and control experiments, each using more than 200 000 monodisperse droplets of 35 pL. CFD simulations of the step emulsification unit, together with experimental characterization, confirm the predicted device performance. The device is prepared by variothermal compression molding. A dual-layer photoresist strategy was used to create the master for replicating the channel systems with two different channel heights and step transitions between them. The core functionality and the applicability of various optical modalities for image-based readout are demonstrated by exemplary biochemical and microbial growth reporter assays. The platform is compatible with automated image-based analysis for high-throughput digital assay readout.

Transcriptional regulators reflect cellular heterogeneity and are key for prognostic modeling. Given the poor prognosis of stomach adenocarcinoma (STAD), regulator-derived signatures are vital for risk stratification. Using multi-stage scRNA-seq data, we delineated the transcriptional regulatory landscape of STAD and identified <i>Helicobacter pylori</i>-associated epithelial heterogeneity in intestinal metaplasia. We then developed a 23-regulator machine learning-based STAD prognostic signature (SPS) from malignant epithelial cells to predict overall survival (OS). Patients with high-SPS exhibited significantly worse OS than patients with low-SPS (hazard ratio [HR] = 1.50, 95% CI: 1.09-2.09, log rank <i>p</i> = 7.11 × 10^-3). Notably, SPS outperformed other established STAD prognosis models across various independent datasets and remained an independent prognostic factor after adjusting for clinical and pathologic factors. Moreover, integrating SPS with tumor stage and age showed superior accuracy to stage alone. Collectively, our study establishes a robust regulator-based prognostic signature, holding potential to facilitate precision prognostication in STAD.

<h4>Introduction</h4>Atrial fibrillation (AF) is the most common and clinically significant arrhythmia requiring treatment. Although an imbalance between the sympathetic and vagal branches of autonomic nervous system (ANS) is known to trigger and sustain AF, the underlying mechanisms remain incompletely understood. Furthermore, there are yet no clinically authorized antiarrhythmic drugs currently targeting the modulation of ANS.<h4>Methods</h4>Targeted LC-MS/MS profiling of plasma 39 neurotransmitters was conducted in a clinical cohort. To identify shared mechanisms, atrial transcriptomes and a sympathetic neuron activation dataset were integrated with DEGs, WGCNA, enrichment, immune infiltration, and machine-learning feature selection, and in silico docking was performed. For <i>in vivo</i> validation, an Ang II-infused mouse model with valsartan intervention was utilized, including transesophageal burst pacing, immunostaining, immunoblotting, and efferocytosis quantification.<h4>Results</h4>Plasma from AF patients exhibited an autonomic-imbalance signature (upregulated monoaminergic and downregulated cholinergic neurotransmitters). Shared ANS-AF bioinformatic analysis highlighted hub genes as CDKN2D, FYTTD1, LRR1, and POPDC3. Immune analyses pinpointed CD47-mediated efferocytosis as a crucial link. <i>In vivo</i>, Ang II induced autonomic remodeling, Ca²⁺-handling suppression, increased apoptosis with CD47/SIRPα upregulation, impaired efferocytosis, and heightened inflammatory signaling; valsartan partially reversed these changes.<h4>Discussion</h4>Multi-omics and <i>in vivo</i> evidence suggests an ANS-immune-atrial remodeling axis in which CD47-SIRPα-dependent efferocytosis blockade is highly associated with atrial inflammation and AF susceptibility, with valsartan acting as a clinically relevant modulator of this pathway.

In almost all aspects of biomedical science, molecular pathology has brought unprecedented value in the diagnosis and management of human disease. Numerous commentators cite haematological disease as the leading genetic cause of global mortality and morbidity, and of these, those of the red blood cells are the most frequent. This narrative review, with a historical perspective, will discuss the role of genetics in these conditions, the leading pathology of red blood cells being the haemoglobinopathies, principally sickle cell disease and thalassaemia, with their many variants, and with potential roles for non-coding RNAs. The impact of genetics into conditions of the red cell cytoplasm will consider the enzymopathies, led by glucose-6-phosphate dehydrogenase deficiency, and extend to those of the cell membrane, causing disease such as hereditary elliptocytosis. Mutations in genes coding almost all the coagulation factors, and several platelet abnormalities, are discussed, as are those linked to conditions of iron overload. Previous, current and evolving technologies for diagnostic testing and their link with potential targeted therapeutic options for patient management are considered.

<h4>Background</h4>The <i>ARFGEF2</i> gene encodes the brefeldin A (BFA)-inhibited GEF2 protein (BIG2), which is distributed in the trans-Golgi network and plays a crucial role in neuronal proliferation and migration during cortical development through its regulation of vesicle transport. Pathogenic mutations in the <i>ARFGEF2</i> gene are associated with autosomal recessive periventricular nodular heterotopia with microcephaly (ARPHM). To date, only slightly more than 20 cases have been reported worldwide. Herein, we presented a case of a patient with West syndrome who was ultimately diagnosed with ARPHM caused by a homozygous variant in the <i>ARFGEF2</i> gene.<h4>Methods</h4>To identify disease-causing mutations, we performed exome sequencing (ES) of a child with West syndrome, and subsequently employed a minigene splicing assay to evaluate the functional impact of the <i>ARFGEF2</i> gene splicing variant.<h4>Results</h4>The patient's clinical manifestations, examination results, treatment, and follow-up course were comprehensively documented. ES revealed a homozygous NM_006420.3: c.5181+1G>T variant in the <i>ARFGEF2</i> gene. Subsequent functional analysis using a minigene splicing assay confirmed that this variant disrupts normal mRNA splicing, causing complete exon 38 skipping and a 118-bp deletion. The translation of this aberrant transcript is predicted to induce a frameshift, resulting in a truncated protein (p.Val1689SerfsTer20).<h4>Conclusion</h4>A novel pathogenic variant was identified by ES, and a minigene splicing assay established its disruptive impact on <i>ARFGEF2</i> mRNA splicing. This study expands the genetic spectrum of <i>ARFGEF2</i> and provides laboratory evidence for clinical diagnosis.

<h4>Background</h4>Addiction is a chronic, relapsing neuropsychiatric disorder despite adverse consequences. Addiction involves neurobiological changes in the reward, motivation, and memory systems, particularly affecting the dopaminergic pathways due to a complex interaction between inherited traits and life experiences.<h4>Methods</h4>To explore the role of genes in addictive behavior, we compiled a list of 332 clinically relevant genes from literature sources and searched GeneAnalytics, STRING web-based programs, and integrated genetic and protein databases for interactions and molecular profiles of genes in tissues and cells, diseases, pathways, biological processes, cellular components, functions, phenotypes, and compounds.<h4>Results</h4>We identified 332 genes associated with addictive behavior and predominantly expressed in the brain associated with schizophrenia, nervous system disorders, and cancer. Nine of the top 10 related pathways showed GPCR protein interaction and signaling involving the <i>CREB1</i>, <i>MAPK1</i>, <i>MAPK3</i>, and <i>AKT1</i> genes. The <i>BDNF</i> gene was recognized in eight of the top 10 high-scoring phenotypes involving the neurotransmitters dopamine and glutamate. We chose to study <i>AKT1</i> as the most identified gene when searching web-based programs and integrated genetic and protein databases. This gene encodes protein kinases that are required for protein phosphorylation, fundamental for most cellular functions, stabilization, regulation of targeted proteins including cell cycle control and signal transduction, neuron formation, and neurological function including the hippocampus. We identified the important PI3/AKT/mTOR signaling pathway that integrates extracellular growth signals, promotes proliferation, and inhibits apoptosis, which is important for brain function. These genes also overlap with other genes associated with ATPase activity, metabolism, and chaperone protein functions that play a role in addiction. In addition, we identified 29 genes that are shared with alcoholism/alcohol use disorder and overlap with interactive molecular functions and processes between alcoholism and addiction.<h4>Conclusion</h4>Our study identified 332 addiction genes from literature sources and used <i>in silico</i> integrated genetic approaches with two searchable web-based programs to provide insights into the complex molecular and genetic architecture of addiction. We identified cell cycle control factors, extracellular growth, signal transduction, and metabolism. The pathways involved selective protein phosphorylation and transduction of proteins as well as neurotrophic factors with mechanisms impacting nervous system development, plasticity and function when disturbed, leading to addictive behavior. Several genes associated with addiction overlapped with those associated with alcohol use disorder.

Lactic acidosis is a strong predictor of mortality in cirrhosis, reflecting both impaired hepatic clearance and systemic tissue hypoxia. We describe a 38-year-old man with decompensated alcohol-associated cirrhosis who developed severe lactic acidosis despite stable hemodynamics and initially nondiagnostic vascular imaging. Repeat computed tomography demonstrated rectal wall thickening with mesenteric fat stranding, concerning for ischemic injury in the setting of a low-flow state. The patient subsequently experienced pulseless electrical activity arrest and died. This case highlights the prognostic significance of rising lactate in cirrhosis and underscores that severe hyperlactatemia should prompt urgent evaluation for occult hypoperfusion even when early imaging is nondiagnostic.

Fasting induces conserved metabolic and redox adaptations that promote stress resistance and longevity. However, the molecular mechanisms linking transient redox changes and altered metabolism to downstream signalling events remain incompletely understood. Using Caenorhabditis elegans, the roles of peroxiredoxins in coordinating redox-dependent responses to fasting and refeeding were determined. A 4-hr fasting protocol over 5 days extended lifespan, improved late-life physiological activity, reduced age-related lipofuscin and lipid accumulation. The fasting protocol generated a transient increase in mitochondrial ROS, promoted mitochondrial turnover, and attenuated age-related mitochondrial fragmentation. These adaptive responses required the activation and nuclear localisation of the stress-responsive transcription factors DAF-16/FOXO and SKN-1/Nrf2. However, these adaptive responses were abolished in prdx-2 and prdx-6 mutant strains, which exhibited persistent redox imbalance, mitochondrial fragmentation, altered stress resistance, and disrupted DAF-16 and SKN-1 signalling. Mechanistically, loss of 2-Cys PRDX-2 impaired activation of the p38 MAPK PMK-1 pathway, resulting in defective SKN-1 activation. In contrast, loss of 1-Cys PRDX-6 disrupted lipid metabolic signalling, preventing induction of NHR-80 and downstream fatty acid desaturases required for metabolic adaptations. Despite distinct initial signalling pathways, both peroxiredoxins converged on the regulation of DAF-16 and SKN-1. Together, these findings identify PRDX-2 and PRDX-6 as redox sensors that translate a fasting-induced transient ROS signature into mitochondrial and lipid remodelling pathways to promote healthy ageing.

Multiple myeloma (MM) develops through asymptomatic precursor stages characterized by progressive remodeling of the bone marrow (BM) immune microenvironment and disruption of bone homeostasis. To delineate changes in natural killer (NK) cell states during disease evolution, we investigated coordinated immune-tumor remodeling by integrating NK cell functional states with plasma cell-intrinsic susceptibility programs derived from CRISPR-based screens across healthy donors (HD), monoclonal gammopathy of undetermined significance (MGUS), smoldering MM (SMM), and newly diagnosed MM patients. The integration of NK cell state-associated gene signatures with plasma cell transcriptional programs revealed stage-specific co-variation between immune and tumor compartments. Public single-cell RNA sequencing datasets were interrogated to resolve NK cell heterogeneity, identifying cytotoxic CD56^dim and regulatory CD56^bright subsets. NK cell dynamics displayed stage-dependent changes, with early expansion followed by the contraction of CD56^dim cells in BM, whereas CD56^bright cells showed predominantly compositional remodeling. Within the CD56^bright subset, transcriptional changes included an increased expression of <i>KLRC1</i> (encoding NKG2A), subsequently validated by multiparametric flow cytometry. In parallel, plasma cell programs associated with NK sensitivity progressively decreased along disease stages, supporting tumor adaptation to immune pressure. The NKG2A ligand HLA-E displayed selective expression within CD16⁺ monocytes and followed a distinct variable pattern across disease stages, highlighting a microenvironmental contribution to NK cell regulation. Collectively, these findings indicate a coordinated process of immune-tumor co-evolution, characterized by dynamic remodeling of NK cell states and plasma cell susceptibility, with the NKG2A-HLA-E axis emerging as a central interface during MM progression.

Three-dimensional (3D) bioprinting has rapidly evolved into a controlling platform for the fabrication of patient-specific biomedical implants, with growing importance in advanced drug delivery systems. Beyond structural tissue engineering, bioprinted constructs now function as programmable therapeutic depots capable of localized, sustained, and stimuli-responsive drug release. This review focuses on recent biomaterial design strategies that enable precise control over drug encapsulation, retention, and release kinetics within 3D bioprinted architectures. The physicochemical and mechanical properties of bioinks, including crosslinking density, porosity, degradation behavior, viscoelasticity, and swelling characteristics, directly influence drug loading efficiency and release dynamics under physiological conditions. The rational tuning of these parameters allows the development of constructs that provide spatially controlled and temporally regulated therapeutic delivery. Recent advances in predictive modeling, such as finite element modeling (FEM), data-driven machine learning approaches, and ML, have significantly improved the ability to correlate material composition, printing parameters, and structural geometry with drug diffusion and degradation-mediated release mechanisms. These tools facilitate the optimization of printing variables including extrusion pressure, nozzle diameter, and layer resolution to ensure structural fidelity while maintaining therapeutic functionality. Emerging strategies incorporating multi-material printing, gradient architectures, and stimuli-responsive biomaterials have expanded the potential of 3D bioprinting for combination therapies and personalized medicine. This review discusses key challenges in translating bioprinted drug delivery systems into clinical applications, including the standardization of drug release characterization methods, and long-term stability assessment.

<b>Background/Objectives</b>: Atrial fibrillation (AF) is a complex polygenic disorder; its genetic architecture remains challenging to fully elucidate. <b>Methods</b>: In this study, we leveraged the extensive genetic overlap between AF and a spectrum of cardiometabolic and behavioral factors-collectively defined by Life's Essential 8 (LE8)-to advance our understanding of its etiology. <b>Results</b>: We first estimated significant genetic correlations between AF and all LE8 components (r_g: -0.11 to 0.19) using LD score regression. We then applied conditional false discovery rate analysis and detected 970 pleiotropic loci associated with AF and at least one LE8 trait. Subsequent colocalization analysis identified 179 loci harboring shared causal variants between AF and one or more LE8 components, which were further refined into 137 distinct colocalized regions. Through region-based annotation and functional predictors, we finally prioritized 164 candidate genes from these colocalized loci, including 40 novel genes. These candidate genes were enriched in pathways related to heart development and regulation of cardiac contraction, and were also enriched among molecular targets of otological agents. Among all LE8 components, blood pressure demonstrated the most extensive shared genetic architecture with AF, supported by the strongest genetic correlation, highest pleiotropic enrichment, and the greatest number of colocalized loci with AF. Polygenic risk scores constructed from these colocalized loci demonstrated significant associations not only for AF but also for arrhythmia and heart failure. <b>Conclusions</b>: Our findings establish a genetic pleiotropy-informed framework that enhances the discovery of novel risk loci of AF and advances our understanding of the shared genetic architecture and potential biological mechanisms between AF and LE8 components.

<h4>Background</h4>The rupture and demyelination of nerve fibers after spinal cord injury (SCI) are primary contributors to neurological dysfunction. The axon attraction signal pathway mediated by netrin-1 is crucial for promoting the effective regeneration and repair of nerve axons. Previous studies have proved that electroacupuncture (EA) can improve nerve function and promote nerve repair and regeneration in rats with SCI, yet its underlying mechanism remains to be elucidated. This study aims to evaluate the effects of EA on neural repair and to investigate its regulatory role in the axon guidance signaling pathway, thereby clarifying the mechanisms by which EA promotes nerve repair following SCI.<h4>Methods</h4>Male Sprague Dawley rats were randomly assigned to the Normal group, Sham group, SCI group, and EA group, with each group further divided into subgroups based on intervention duration: 7, 14, and 28 days. Allen's method was employed to establish the SCI model. Dazhui (GV14) and Mingmen (GV4) acupoints were selected for EA intervention. Basso-Beattie-Bresnahan (BBB) score and the inclined plate test were used to evaluate the motor function of rats in each group. Morphological and structural changes in the injured spinal cord were assessed through magnetic resonance imaging (MRI). The fractional anisotropy (FA), radial diffusivity (RD), and mean diffusivity (MD) values in the injured area were measured via diffusion tensor imaging (DTI), and the morphological changes of nerve fiber bundles were analyzed via diffusion tensor tractography (DTT). The expressions of netrin-1, deleted in colorectal cancer (DCC), ras-related C3 botulinum toxin substrate 1 (Rac1) and F-actin were quantified using immunofluorescence (IF) staining, Western blot (WB) and real-time quantitative polymerase chain reaction (RT-PCR).<h4>Results</h4>Compared with the Sham group, the SCI group exhibited significant deficits in motor function (P<0.01), disorganized spinal cord tissue structure, a markedly increased lesion area (P<0.01), impaired integrity of nerve fiber bundles, a significant decrease in FA value (P<0.01), and significantly elevated MD and RD values (P<0.01). Moreover, the expression levels of netrin-1, DCC, Rac1, and F-actin were significantly reduced (P<0.01). EA improved the behavioral performance of rats with SCI (P<0.01). Following EA intervention, the extent of spinal cord structural damage was alleviated, with increased FA values, decreased MD and RD values (P<0.01), alongside evident repair and reconstruction of damaged fiber bundles. Additionally, EA upregulated the expression levels of netrin-1, DCC, Rac1, and F-actin (P<0.01).<h4>Conclusions</h4>EA can significantly improve the motor function of rats with SCI, regulate the nerve guidance factor netrin-1 and its receptor DCC, as well as the molecular switch Rac1, thereby promoting axonal cytoskeletal remodeling and facilitating neural repair. The nerve repair effect of EA may be achieved by regulating the axon attraction signal pathway mediated by netrin-1.

Identifying the genetic changes that shaped recent human adaptation depends on our ability to detect selection from genomic data. Summary statistics from haplotype scans have been widely used for that purpose, aggregating genetic signal over windows, though resolution is limited by linkage and their power may diminish as sweeps approach fixation, as in the case of the integrated haplotype score (iHS). Ancient DNA based scans recover signal by analysing time-series trajectories, but the majority of human populations fall outside the geographic range of any existing ancient DNA dataset. Pairwise coalescence times provide a way to complement statistics and can be applied to any modern cohort, yet computing them densely enough at cohort scale poses a computational challenge due to the quadratic growth in the number of haplotype pairs. We introduce gamma_smc_cu , a GPU implementation of the Gamma-SMC algorithm (Schweiger and Durbin, 2023) for pairwise time-to-the-most-recent-common-ancestor (TMRCA) inference. Applied to the 1000 Genomes Project (3,202 phased samples, corresponding to 6,404 haplotypes; 829,638 within-population pairs across 26 populations and five different continental ancestries; ∼10 12 per-site posterior evaluations), it yields a gene-level TMRCA landscape of 17,823 autosomal protein-coding genes after masking for segmental duplications. The scan recovers well-known sweeps ( LCT, SLC24A5, EDAR, FADS1, HERC2, ABCC11 ) and, combined with a depleted-to-enriched variant-class profile, resolves haplotype-block signals down to the gene level. Of seven case studies, two are developed in the main text — GRK2 / ADRBK1 (chr11q13.2; SAS+EUR) and TREML1 / TREM2 (chr6p21.1) — and the remaining five ( IFIH1 chr2q24/IBS, CCDC92 chr12q24/CDX, SLC6A15 chr12q21/CHS, BPIFA2 chr20q11/GIH, CLEC6A chr12p13/CDX) are presented in the Supplementary Information (SI). Notably, TREML1 / TREM2 is a shared out-of-Africa signal — ranked below the within-population 1% tail in 16 of 19 non-African 1000 Genomes panels that PopHumanScan and five landmark haplotype-based scans miss. A previous 10 kb-windowed-mean iHS scan dilutes the cluster of extreme sites packed inside the ∼5 kb gene bodies, while our own gene-level iHS independently recovers the locus in three South Asian panels (BEB, STU, ITU; top 0.4% genome-wide). We cross-validate the seven cases against the 9.7 million per-variant selection posteriors from a recent West-Eurasian ancient DNA scan. BPIFA2 is detected concordantly ( s ≈ 1.8% per generation). GRK2 and CCDC92 reach detection threshold in flanking variants but not within their own gene bodies, while the TREML1 / TREM2 cluster falls below it. To calibrate novelty, we review the candidate landscape against an expanded eight-catalog set spanning curated haplotype scans, the largest current West-Eurasian ancient-DNA leads, and a recent 26-population iHS refinement; the vast majority of our loci overlap at least one prior entry, and only a handful — including TREML1 / TREM2 — remain unflagged. The contributions of this work are gene-level resolution, systematic ancient DNA cross-validation, and a reusable TMRCA landscape that complements aDNA panels.

<h4>Background</h4>Although inhibitors of mRNA translation are being evaluated as anti-cancer agents, the dynamics of protein synthesis throughout tumour progression are still poorly understood. Here we assess how alterations in mRNA translation during early tumorigenesis affect tumour development in KRAS-driven lung adenocarcinoma (LuAd).<h4>Methods</h4>We deployed autochthonous mouse models of LuAd driven by oncogenic KRAS^G12D combined with moderate overexpression of MYC and simultaneously manipulated mRNA translation by deleting the mRNA helicases eIF4A1 and eIF4A2 or by administering pharmacological inhibitors of protein synthesis, such as rapamycin. This permits synchronous assessment of LuAd initiation and progression in vivo and is amenable to parallel ex vivo culture of tumour-derived cells for detailed analysis of protein synthesis (using ribosome footprinting) and metabolic landscapes. These approaches also allowed us to perform multiplex imaging and spatial transcriptomics to characterise tumour formation in altered mRNA translation conditions and to compare results obtained in mice against the Lattice-A cohort of non-small cell lung cancer (NSCLC) patients.<h4>Results</h4>Deletion of the mRNA-translation repressor, eIF4A2 in KRAS-driven LuAd leads to a dysregulated protein synthesis landscape characterised by a strongly upregulated secretome, enlarged secretory compartments, increased oxidative metabolism and acquisition of senescence-like characteristics. Paradoxically, this overdriven secretory protein synthesis landscape delays tumorigenesis and leads to the appearance of clusters of non-proliferative, p21-positive KRAS^G12D-expressing cells in the lung. Consistently, reduction of mRNA translation with rapamycin in Eif4a2-deleted tumours suppresses senescence and restores tumorigenesis. Importantly, some Eif4a2 knockout cells overcome senescence to form tumours that exhibit enhanced MAP-kinase signalling and, in contrast to eIF4A2^+/+ lesions, these were eradicated by administration of a MEK inhibitor. Consistently, MAP-kinase signalling was significantly increased in human NSCLC expressing low levels of eIF4A2.<h4>Conclusions</h4>Our study highlights that restraint of mRNA translation by eIF4A2 is critical in the early-stages of KRAS-driven LuAd to allow bypass of oncogene-induced senescence and tumour progression. Importantly, because tumours with dysregulated mRNA translation rely heavily on MAP-kinase signalling they are exquisitely sensitive to MEK inhibition, and this indicates the possibility that low expression of eIF4A2 could be used to identify potential responders to MEK inhibitors in clinical trials.

<h4>Aim of the study</h4>To evaluate the diagnostic accuracy of the Warsaw Cognitive Assessment Scale (Warszawska Skala Oceny Poznawczej, WSOP) for detecting mild cognitive impairment (MCI) and dementia, and to compare its performance with the Addenbrooke's Cognitive Examination III (ACE-III) and the Mini-Mental State Examination (MMSE).<h4>Clinical rationale for the study</h4>Early detection of cognitive impairment remains challenging in clinical practice. Existing screening tools are limited by administration time, sensitivity to early deficits, and reliance on auditory input. The WSOP was developed as a brief, multidomain screening instrument tailored to the Polish population, aiming to balance diagnostic accuracy with clinical efficiency.<h4>Material and methods</h4>A total of 370 participants (111 Controls, 138 with MCI, 121 with mild dementia) were assessed using WSOP, ACE-III, and MMSE. Group differences were analyzed using non-parametric tests, and convergent validity was evaluated with Spearman's correlations. Diagnostic accuracy was assessed using receiver operating characteristic (ROC) analysis, including area under the curve (AUC), sensitivity, specificity, and optimal cut-off values. Additional analyses controlled for age, education, and sex.<h4>Results</h4>WSOP demonstrated high diagnostic accuracy comparable to ACE-III. For distinguishing Controls from MCI, AUC values were 0.945 (WSOP) and 0.951 (ACE-III), both exceeding MMSE (0.80). For MCI versus dementia, AUC values were 0.943 and 0.964, respectively. The optimal WSOP cut-offs were 67.5 for MCI and 53.5 for dementia. Diagnostic performance remained stable after adjusting for demographic variables. WSOP scores showed moderate to strong correlations with ACE-III, providing initial evidence of convergent validity.<h4>Conclusions and clinical implications</h4>WSOP is a brief, reliable multidomain cognitive screening tool with diagnostic accuracy comparable to ACE-III. Its balance of comprehensiveness and efficiency supports its potential use in clinical screening settings, pending further validation in broader populations.

The immunosuppressive tumor microenvironment, characterized by impaired antigen presentation, "don't eat me" signaling, and T-cell exhaustion, challenges effective cancer immunotherapy. Conventional T-cell-redirecting bispecific engagers (BiTEs) show limited efficacy in solid tumors because of poor T-cell infiltration, rapid clearance, and dose-limiting cytokine release associated with the CD3 agonism. Therefore, we aimed to enhance macrophage-mediated tumor clearance and T-cell priming by developing a bispecific macrophage nanoengager (nanoBIME) that integrates dual immune checkpoint blockade (anti-SIRPα and anti-PD-L1) with STING-agonist delivery (SR717) in a single poly(lactic-<i>co</i>-glycolic acid) nanocarrier (BIS-PLGA-NP@SR717). BIS-PLGA-NP@SR717 exhibited uniform nanoscale features, efficient SR717 encapsulation, and stable dual-antibody conjugation, enabling systemic delivery with sustained intracellular release. Dual-ligand presentation markedly enhanced receptor binding and nanoparticle uptake in macrophages and PD-L1-expressing tumor cells. BIS-PLGA-NP@SR717 synergistically promoted macrophage phagocytosis, repolarized tumor-associated macrophages toward an M1 phenotype, enhanced dendritic cell maturation, and CD8⁺ T-cell activation; additionally, it elevated granzyme B, perforin, and IFN-γ production, resulting in robust T-cell-dependent tumor cell killing. In CT26 tumor-bearing mice, BIS-PLGA-NP@SR717 achieved superior tumor accumulation, deep intratumoral penetration, and dose-dependent tumor regression with minimal systemic toxicity. Transcriptomic profiling further revealed the coordinated activation of phagosome-related pathways, antigen presentation, STING/IFN pathways, chemokine signaling, and T-cell receptor engagement. Collectively, BIS-PLGA-NP@SR717 represents an integrated, multiaxis immunotherapeutic nanoplatform that synchronizes innate activation, macrophage remodeling, and T-cell effector functions. Its modular architecture provides a versatile framework for next-generation immunotherapies for solid tumors.

Mitochondrial pyruvate carrier (MPC) inhibition was found protective in models of neurodegenerative diseases, such as Alzheimer's and Parkinson's. However, little is known about MPC as a potential therapeutic target in Huntington's disease (HD), a neurodegenerative disorder with dysregulation of the pro-survival pathway integrated stress response (ISR). Here, we investigate if MPC inhibition modulates the ISR and mitigates mutant huntingtin (mut-Htt) proteotoxicity in a cellular HD model. We treated cells expressing N-terminal fragments of wild-type- (wt-) or mut-Htt with two MPC inhibitors (mitoglitazone and UK5099) or solvent control. Metabolism was assessed analysing resazurin reduction, oxygen consumption, extracellular acidification, and ATP levels. ISR activation and huntingtin proteostasis were assessed using western-blot and filter-trap assays. Mut-Htt-expressing cells showed decreased resazurin reduction and ATP levels, and increased eIF2α phosphorylation, indicating metabolic stress and ISR activation. MPC inhibitors (100 µM) increased resazurin reduction and decreased respiration. The latter was rescued by the membrane-permeant methyl pyruvate, which bypasses MPC inhibition. In wt-Htt-expressing cells, MPC inhibitors increased levels of ATP and ISR markers, suggesting metabolic adaptation and ISR activation. In mut-Htt-expressing cells, MPC inhibitors preserved ATP levels and attenuated mut-Htt-induced eIF2α phosphorylation but without changing soluble or aggregated mut-Htt levels. This work showed that MPC inhibition differentially modulates the ISR: it activates ISR in control cells and attenuates overactive ISR in mut-Htt-expressing cells. However, MPC inhibition did not impact the proteostasis of N-terminal fragment mut-Htt. Further studies are essential to explore MPC inhibition in less severe full-length mut-Htt-expressing models to better understand its therapeutic potential in HD.

<h4>Background</h4>Neurodegenerative and psychiatric disorders, including Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), and schizophrenia (SZ), are characterized by progressive neuronal loss and synaptic dysfunction. Despite their severity, effective disease-modifying treatments remain unavailable, largely due to the elusive nature of their underlying molecular mechanisms.<h4>Methods</h4>To elucidate these mechanisms, we conducted an integrative systems biology analysis incorporating transcriptomic datasets, in silico proteomic networks, and inferred metabolomic profiles. Machine learning (ML) and deep learning (DL) models were employed to identify regulatory networks associated with oxidative stress, immune response, and synaptic signaling. Furthermore, network pharmacology approaches were applied to explore multi-target intervention strategies using bioactive compounds from traditional Chinese medicine (TCM).<h4>Results</h4>Our integrative analysis revealed extensive overlap in dysregulated biological processes across all four disorders, particularly involving oxidative stress and immune activation. We identified TP53, NFE2L2, and PPP3CA as central regulatory hubs driving these pathologies. Notably, computational predictions highlighted that TCM-derived compounds, specifically stigmasterol and dodecanoic acid, exhibit promising multi-target effects for modulating these oxidative and inflammatory responses. Subsequent in vivo experimental validation was performed exclusively to corroborate the disease-associated pathways and core gene dysregulation in an Aβ₄₂-induced AD model. These findings demonstrated molecular and behavioral phenotypes consistent with our multi-dimensional computational predictions, establishing a robust mechanistic rationale that merits future in vivo pharmacological validation for the predicted bioactive compounds.<h4>Conclusion</h4>This study highlights the utility of a multi-disease, multi-dimensional framework in uncovering shared pathogenic signatures. By integrating computational analytics with pharmacological modeling and experimental validation, we identified key regulatory genes and natural compounds with therapeutic potential. These findings provide a theoretical foundation for the development of multi-target, personalized treatment strategies against neurodegeneration.

Cancers reflect aberrant growth and differentiation of normal cell populations. Biological understanding of small intestine neuroendocrine tumors (SI-NETs) is hampered because their closest normal counterparts, enteroendocrine cells (EECs), constitute tiny fractions of intestinal epithelium. Recent characterization of adult human EEC ontogeny from intestinal stem cells can help overcome that limitation. Transient expression of the transcription factor gene ASCL1 normally ensures proper timing and fidelity of well-differentiated EECs, which express NEUROD1. Here, we report that SI-NETs resembled mature enterochromaffin cells; however, individual tumor cells coexpressed stem/progenitor genes, harboring each differentiation state along the EEC trajectory except ASCL1+ precursors. We found that enhancers normally active, and others inactive, during EEC differentiation underlie aberrant SI-NET gene activity. SI-NETs uniformly expressed NEUROD1 but lacked ASCL1, owing to inaccessible chromatin and repressive H3K27me3 marking at the ASCL1 locus. Multiple cyclin-dependent kinase inhibitor (CDKi) genes were similarly silenced, other than CDKN1B, the only gene recurrently mutated in SI-NETs. Deletion of CDKN1B altered cell cycle kinetics during human EEC differentiation, and deletions of ASCL1 or CDKN1B activated certain genes that are expressed in SI-NETs but not in the normal EEC trajectory. We propose that a limited CDKi repertoire and absence of ASCL1-dependent constraints on EEC maturation together explain unique SI-NET characteristics.

Intensive care units (ICU) patients are highly vulnerable to inaccurate drug dosing. Pharmacogenomics (PGx) studies the role of inherited genetic variation in drug metabolism and dose efficacy. To assess the prevalence of PGx variants that may influence therapeutic effect in the ICU, we carried out whole genome sequencing (WGS) of 210 Qataris in ICU care at Hamad Medical Corporation (HMC), Doha, Qatar and assessed the WGS for predicted deleterious variants of genes that metabolize 30 drugs commonly prescribed in the ICU. PGx variation was evaluated using two complementary approaches. First, variants with established functional interpretation were assessed using CPIC guidelines and star-allele haplotypes inferred by PharmCAT to estimate the prevalence of alleles associated with abnormal drug metabolism. Second, a broader exploratory analysis examined computationally predicted deleterious single-nucleotide variants in pharmacogenes that currently lack CPIC guidelines or defined star alleles, with these findings interpreted as descriptive of genomic variation rather than clinical metabolizer phenotypes.Of the ICU patients that received the 5 most commonly prescribed drugs (warfarin, phenytoin, midazolam, vancomycin, levetiracetam), 93% had deleterious metabolism-related variants. Ninety-one % of ICU patients carried at least one variant in a gene with known PGx relevance that could potentially impact the metabolism or activity of at least one medication they received. Most patients had ≥14 deleterious variants of genes that affect the metabolism of administered drugs. Comparison of the deleterious variants related to metabolism of ICU drugs with African/African American and European populations revealed significant population specificity in ICU related PGx variants. Together, these data suggest that population specific, PGx based on the individual's genome likely plays a significant role in effective, safe dosing in the ICU setting.

This study focused on the synthesis of a novel series of (4-bromothiophen-2-yl)methyl 1-naphthoate derivatives (5a-5h) <i>via</i> the Suzuki-Miyaura cross-coupling with moderate to excellent yields. Acute pentylenetetrazol (PTZ) and 6 Hz psychomotor seizure models were used to investigate <i>in vivo</i> anticonvulsant effects. The 5b, 5d, and 5h showed significant outcomes concerning mortality, protection, and seizure severity. In addition, the goal of the current study was to use a multifaceted strategy that integrates molecular docking (AutoDock Vina), <i>in silico</i> ADMET (SwissADME, Molsoft), and network pharmacology to explore their potential as anti-epileptic drugs against important targets (GABAA receptor (8G5G), SV2A (3O7P, 1PW4)). To find common targets, we built PPI networks and carried out functional enrichment analysis. As compared to standard medications (levetiracetam, diazepam, and PTZ), all compounds (5a-5h) showed greater binding affinities (-7.6 to -9.7 kcal mol^-1). ADMET profiles revealed drug-like characteristics with good bioavailability scores (0.55), but they also highlighted P-gp substrate liabilities and low solubility as important improvement considerations. Molsoft web server highlighted their good BBB permeation. By focusing on hub genes (SRC, AKT1, MAPK1/3, STAT3, EGFR) implicated in key signaling pathways such as PI3K-Akt, MAPK, JAK-STAT, and GABAergic synapse signaling, network analysis showed that these drugs had 15-25 targets in common with epilepsy.

<b>Background/Objectives</b>: Metabolic dysfunction-associated steatohepatitis (MASH) is frequently accompanied by disturbances in coagulation and metabolic homeostasis, partly related to impaired handling of vitamin K-dependent pathways. Although vitamin K is often administered to correct abnormal coagulation tests, its biochemical impact in hospitalized patients with MASH remains insufficiently characterized. This study aimed to evaluate changes in coagulation and metabolic parameters in hospitalized patients with MASH receiving vitamin K supplementation. <b>Methods</b>: We conducted a retrospective study of 84 hospitalized MASH patients who received vitamin K supplementation. Biochemical parameters were recorded at admission and discharge to assess short-term changes during hospitalization. <b>Results</b>: Vitamin K supplementation was associated with modest changes in coagulation parameters, including reductions in PT, INR, and aPTT (e.g., PT decreased from 13.00 s to 11.00 s). Small numerical changes in transaminases, fasting glucose, and total cholesterol were observed during hospitalization, with limited clinical relevance. These patterns were comparable across fibrosis stages, with no significant differences between groups. <b>Discussion</b>: The observed biochemical findings are likely in-hospital factors rather than a direct metabolic effect of vitamin K. <b>Conclusions</b>: Vitamin K supplementation was associated with modest changes in coagulation parameters and small, clinically negligible variations in selected metabolic markers in patients with MASH, irrespective of fibrosis stage. These findings suggest a supportive biochemical effect in selected contexts; further prospective studies are needed to clarify their clinical relevance.

Differentiation therapy holds significant potential for the treatment of multiple myeloma (MM). We previously identified that the aminopeptidase N (APN) inhibitor Bestatin promotes MM cell differentiation. Herein, we elucidate the underlying molecular mechanisms of this process. Utilizing MM1.S, U266, and RPMI-8226 cell lines, a combination of CCK-8 assays, flow cytometry, Wright-Giemsa staining, Western blotting, qRT-PCR, ELISA, APN enzymatic activity analysis, SA-β-gal staining, and bioinformatic analyses revealed elevated APN expression across all cell types. Bestatin treatment induced MM cell differentiation in a concentration-dependent manner, which was accompanied by the upregulation of the differentiation marker CD49e, increased immunoglobulin light chain secretion, elevated cellular senescence, and a concomitant suppression of cell proliferation and APN enzymatic activity. Mechanistically, Bestatin exerts its effects by downregulating the CD79B/BTK signaling pathway, thereby activating the downstream transcription factor STAT3. Consistent with this axis, direct inhibition of CD79B/BTK alone was sufficient to induce differentiation, while blockade of STAT3 completely abrogated the differentiation-promoting effect of Bestatin. The APN-neutralizing antibody (WM15) yielded consistent results with Bestatin, further validating this regulatory axis. Furthermore, both the CD79B/BTK inhibitor Ibrutinib and the STAT3 agonist GCDA potentiated the cytotoxicity of the clinical MM drug Ixazomib. Bestatin itself synergized with Ixazomib and enhanced the anti-proliferative effect of IL-6. In summary, our findings establish that the APN inhibitor Bestatin induces MM cell differentiation via the CD79B/BTK-STAT3 signaling axis. Targeting this pathway represents a promising strategy to enhance the efficacy of Ixazomib, providing a compelling rationale for novel combination therapies in MM.

<b>Background/Objectives</b>: Childhood interstitial lung disease (chILD) represents a heterogeneous group of rare pulmonary disorders. Practical diagnostic approaches tested for feasibility and impact in comprehensive cohorts are lacking. We aimed to assess a simple etiologically focused classification approach, clarify the role of genetic testing and quantify the impact of non-pulmonary organ manifestations. <b>Methods</b>: We hypothesized that chILD can be classified in a clinically meaningful and versatile way by answering three questions: Which children have an etiological chILD diagnosis due to (1) identified (exposure-related) cause/lung injury, or (2) systemic disease? (3) In how many children without an etiological diagnosis can a genetic cause be identified? We also calculated the predictive value of non-pulmonary organ involvement for underlying systemic conditions. <b>Results</b>: Among 1693 patients, 24.7% were grouped as ILD related to exposure, 22.7% as ILD with systemic condition, 16.6% as ILD with genetic diagnosis of systemic disease, 10.0% as ILD with genetic diagnosis affecting the lungs only, and 25.8% as ILD without genetic diagnosis. The average genetic diagnostic yield was 50.8%, with higher rates in interstitial pneumonia (61.4%) or pulmonary alveolar proteinosis (87.1%). The presence of ≥two non-pulmonary organ manifestations increased the likelihood of an underlying systemic disease by three to five-fold. <b>Conclusions</b>: An etiological diagnostic strategy effectively classifies chILD and guides genetic testing. Exome or genome sequencing should be considered if ≥two non-pulmonary organs are involved or if the initial diagnosis becomes uncertain due to an unusual disease course or signs of a second underlying condition.

<h4>Background</h4>The precise identification of cancer subtypes through the integration of multi-omics data has emerged as a key research direction in bioinformatics. Among existing multi-omics integration methods, similarity graph-based clustering algorithms have attracted widespread interest owing to their capacity to effectively characterize the association patterns between samples. However, the majority of existing methods primarily focus on first-order relationships among samples while ignoring the prevalent high-order neighborhood relationships, and fail to fully exploit the complementary information from different omics.<h4>Methods</h4>To address these limitations, we propose an innovative multi-omics integration framework termed MHSGTR, which integrates multi-omics data by combining Motif high-order similarity graphs and tensor regularization to identify cancer subtypes. Specifically, MHSGTR introduces Motif theory to construct a high-order similarity graph and designs a high-order graph learning term to obtain a hybrid similarity that integrates both first-order and high-order information, thereby capturing the latent high-order structural information among samples. For multi-omics data integration, we employ third-order tensor regularization constraints to explore complementary information across multi-omics data, coupled with an attention module to adaptively learn omics-specific weights for constructing a consensus similarity graph. Final clusters are derived via spectral clustering.<h4>Results</h4>Comprehensive experiments on eight TCGA cancer datasets and a case study on adrenocortical carcinoma (ACC) demonstrate that MHSGTR achieves superior clustering performance and identifies cancer subtypes with significant biological differences, showcasing its effectiveness in robust multi-omics integration.

The pancreas is an organ with two functions: endocrine and exocrine. The proper functioning of the pancreas depends on many factors. One of these is trace elements-precise control of trace element homeostasis is important for both the endocrine and exocrine parts. This review provides a comprehensive summary of current knowledge regarding the role of trace elements: iron (Fe), copper (Cu), cobalt (Co), iodine (I), manganese (Mn), zinc (Zn), silver (Ag), cadmium (Cd), mercury (Hg), lead (Pb), and selenium (Se) in pancreatic physiology and their influence on the pathogenesis of key diseases of this organ, such as diabetes (DM), acute (AP) and chronic pancreatitis (CP), autoimmune pancreatitis (AIP), and pancreatic cancer (PC). Trace elements, including Fe, Cu, Zn, Se, and Mn, play a fundamental role in maintaining endocrine and exocrine homeostasis, participating in insulin synthesis, stabilizing digestive enzymes, and the functioning of antioxidant systems. It has been demonstrated that disturbances in their concentrations lead to the activation of pathological molecular pathways, including oxidative stress, chronic inflammation, and beta-cell apoptosis. In the context of diabetes, excess Fe promotes ferroptosis, whilst exposure to heavy metals such as Cd, Pb, and Hg induces insulin resistance and pancreatic islet dysfunction. In the course of pancreatitis, elements such as Zn and Se exhibit protective potential by stabilizing tissue barriers, whereas toxic metals impair ion transport, exacerbating fibrotic processes. Furthermore, analysis of available data indicates a significant association between heavy metal accumulation and pancreatic carcinogenesis, driven by DNA damage and oncogene modulation. Understanding pancreatic metallomics opens new prospects for early diagnosis, environmental prevention, and the development of targeted therapeutic strategies that restore the body's micronutrient balance.

Prostate-specific membrane antigen (PSMA) ligands are currently one of the popular platforms for creating drugs targeted at prostate cancer tumor cells. In this review, we have collected and systematized current approaches to the synthesis of PSMA ligands based on N-[N-[(<i>S</i>)-1,3-dicarboxypropyl]carbamoyl]-(<i>S</i>)-L-lysine (DCL), the most widely used scaffold in the design of such ligands. The main approaches to each stage of the synthesis of PSMA inhibitors with various structures are considered in detail, and the existing synthetic techniques are compiled and analyzed. We also review examples of using different synthetic pathways for diagnostic and therapeutic drugs based on PSMA ligands. We propose an algorithm for selecting a synthetic route based on the structure of the target ligand.

Hyaluronic acid (HA) hydrogels have attracted increasing interest for biomedical applications due to their tunable properties and biocompatibility. <b>Methods:</b> In this study, hyaluronic acid HA-based hydrogels were developed using two distinct crosslinking strategies: physical crosslinking through poly(vinyl alcohol) (PVA) incorporation and covalent crosslinking via DCC/NHS-mediated reactions. Piroxicam (Px) was included as a model drug to evaluate the drug delivery potential of the resulting systems. The hydrogels were characterized in terms of morphology, swelling behaviour, adhesion, enzymatic degradation, drug release, and in vitro cytocompatibility. <b>Results:</b> The results indicate that formulation parameters significantly influence the overall performance of the systems. PVA-containing hydrogels exhibited higher swelling capacity and improved adhesive properties, while covalently crosslinked networks showed reduced swelling and enhanced structural stability and resistance to enzymatic degradation. Drug release profiles were dependent on network structure, with more compact systems displaying slower release behaviour. In vitro assays suggested that the developed hydrogels are cytocompatible and that drug incorporation influences both release kinetics and cellular response. However, it should be noted that the biological evaluation was performed under simplified in vitro conditions, which primarily reflect specific aspects such as cell viability and migration. <b>Conclusions</b>: This study provides a comparative analysis of physical and covalent crosslinking strategies within a HA platform and highlights how formulation variables influence key physicochemical and biological properties. These findings contribute to the rational design of HA-based hydrogels, although further studies are required to establish their performance in more complex biological environments.

Periventricular nodular heterotopia (PVNH) is a genetically heterogeneous malformation of cortical development with variable neurological outcomes. Among recessive forms, <i>ARFGEF2</i>-related disorder is uniquely characterised by the association of diffuse PVNH and progressive microcephaly. We describe a two-year-old boy born to consanguineous parents who presented with severe developmental delay, hypotonia, progressive microcephaly, and infantile-onset epileptic spasms with developmental regression. Brain MRI showed extensive bilateral PVNH associated with callosal hypoplasia and ventriculomegaly. EEG revealed dysmature background activity with multifocal epileptiform discharges and runs of asynchronous fast activity during sleep. Genetic testing identified a novel homozygous nonsense variant in <i>ARFGEF2</i>. The clinical course was characterised by drug-resistant epilepsy and multisystemic involvement, including feeding difficulties and recurrent respiratory infections. To contextualise this case, we performed a comprehensive review of previously reported patients, further delineating the clinical, neuroradiological, and electroclinical spectrum of <i>ARFGEF2</i>-related disorder. This case highlights progressive microcephaly as a key distinguishing feature of <i>ARFGEF2</i>-related PVNH and underscores the importance of early genetic diagnosis to guide targeted surveillance for extra-CNS complications and multidisciplinary care.

Neurodegenerative diseases (NDDs) lack effective disease-modifying therapies. The FOXO transcription factors serve as integrative hubs of cellular stress responses, operating through cell-autonomous homeostasis, intercellular coordination, intracellular quality control, and cell fate decisions four hierarchical tiers. This review systematically examines isoform-specific functions: FOXO1 governs metabolic reprogramming and mitochondrial biogenesis; FOXO3 acts as the principal oxidative stress sensor with context-dependent neuroprotective or pro-apoptotic outputs; FOXO4 regulates cellular senescence; and FOXO6 maintains synaptic metabolic support. These functions vary across cell types and disease stages, with post-translational modifications determining functional transitions. FOXO proteins participate in complex interactions with disease-specific pathological proteins, either promoting clearance and repair or exacerbating neurodegeneration depending on stress intensity and chronicity. Therapeutic strategies targeting FOXO remain in preclinical and early clinical stages. Key challenges include disease stage-dependent dosing, cell-type-specific delivery, blood-brain barrier penetration, and metabolic side effects. Future directions emphasize biomarker-guided patient stratification and precision interventions aligned with the spatiotemporal dynamics of FOXO signaling. Unlike prior reviews focusing on single pathways or diseases, this work integrates isoform-specific, stage-dependent, and cell-type-resolved FOXO functions into a unifying hierarchical framework.

Diabetes mellitus compromises bone regeneration and increases implant failure rates, posing persistent challenges in orthopedic and dental therapies. Although macrophage-centered immunomodulation has been widely investigated, the role of neutrophils, particularly under diabetic conditions, remains insufficiently defined. Here, we identify neutrophil dysfunction, characterized by impaired chemotaxis and elevated oxidative stress, as a critical upstream driver of impaired osseointegration in diabetes. We demonstrate that dietary flavonoid quercetin restores neutrophil redox homoeostasis in association with reactivation of Nrf2/HO-1 signaling, attenuates pathological neutrophil extracellular trap formation, and promotes MerTK-associated efferocytosis by macrophages. Efferocytosis of quercetin-treated neutrophils subsequently reprograms macrophage toward an anti-inflammatory, pro-regenerative M2-like phenotype, therefore establishing a microenvironment favorable for angiogenesis and osteogenesis. Based on these insights, we engineer a biodegradable tri-layered coating on polyetheretherketone (PEEK) implant, comprising an osteoconductive nano-hydroxyapatite base layer, a quercetin-rich antioxidant intermediate layer, and an outermost surface functionalized with the chemotactic peptide formyl-methionyl-leucyl-phenylalanine (fMLP) to actively recruit neutrophils. In a diabetic rat model, this rationally designed interface enables temporal coordination of innate immune responses, leading to substantially improved vascularized bone regeneration and implant osseointegration. These findings highlight neutrophils as key therapeutic targets in diabetic bone repair and propose an immunomodulatory biomaterials strategy that addresses upstream immune dysregulation in compromised metabolic conditions.

Molecular glue degraders (MGDs) are an emerging class of small molecules that promote selective protein degradation by inducing neomorphic interactions between E3 ubiquitin ligases and non-native substrates, referred to as neosubstrates. Clinically validated examples include thalidomide analogs that recruit cereblon (CRBN) to degrade IKAROS family zinc finger 1/3 in multiple myeloma, and arylsulfonamide-based MGDs that promote the degradation of RNA-binding protein 39 in acute myeloid leukemia and solid tumors. These molecules highlight the therapeutic potential of this modality in oncology. These findings underscore the promise of MGDs for eliminating oncogenic proteins previously considered undruggable and overcoming resistance to conventional inhibitors. Despite these successes, the current MGD landscape relies heavily on a limited set of E3 ligases-mainly CRBN which constrains substrate diversity, tissue selectivity, and durability of clinical response. Expanding the therapeutic utility of MGDs requires the systematic identification of novel ligases and their neosubstrates, accompanied by a deeper understanding of the mechanistic basis of ligase-substrate recognition. Recent technological advances, including chemoproteomics, ubiquitin-remnant profiling, degron mapping, clustered regularly interspaced short palindromic repeats-based functional genomics, and artificial intelligence-driven structural modeling, are advancing the discovery of new ligase-substrate pairs and enabling the rational design of degraders. Parallel progress in next-generation CRBN E3 ligase modulators, noncanonical MGDs, and structure-guided engineering further illustrates the expanding therapeutic versatility of this approach. By integrating multidisciplinary discovery strategies with translational oncology, the field is moving toward the development of next-generation MGDs with enhanced specificity, broader substrate scope, and improved resistance profiles. This study aims to elucidate how these innovations expand the degradable proteome and establish MGDs as a cornerstone of precision cancer therapy, thereby redefining the boundaries of drug discovery and providing customizable degraders tailored to diverse cancer contexts.

Despite the placement of millions of dental implants annually, one-quarter to one-half of cases require concurrent guided bone regeneration (GBR), where a recent meta-analysis of 100 studies reported an approximately 26% complication rate that exposes the structural and biological limits of current barrier membranes. Conventional collagen and PTFE-based membranes function predominantly as passive occlusive barriers, lacking the bioactivity, controllable degradation, and immunomodulatory capacity demanded by the dynamic alveolar microenvironment, in which up to 50% of ridge width can be lost within 12 months post-extraction. Addressing this gap requires reframing GBR membranes as programmable interfaces. This review presents a three-lens framework - alveolar-bone-specific osteoimmune biology, metabolically active and stimuli-responsive material platforms, and translational bottlenecks - to systematically chart the field. We synthesize recent advances across four next-generation material families (polymer composites, biodegradable Mg/Zn alloys, MXene-based systems, and citrate-based polymers), four hierarchical structural strategies (bilayer, Janus, gradient, and 4D-printed architectures), and complementary functionalization strategies that include surface chemistry tailoring, bioactive ion release, and stimuli-responsive triggers, explicitly mapping how each modulates mechanical retention, degradation kinetics, antibacterial activity, and macrophage M1-to-M2 polarization. We further evaluate the clinical performance of these material platforms in alveolar ridge augmentation and identify platform-specific post-market follow-up endpoints required for regulatory translation. Finally, we articulate six open mechanistic questions and a near-term agenda integrating artificial intelligence/machine learning-driven design, microfluidic oral-microenvironment models, and standardized large-animal alveolar protocols. This framework repositions GBR membranes toward programmable, osteoimmune-active regenerative platforms, charting an actionable path from bench to clinic.

<h4>Background</h4>Di (2-ethylhexyl) phthalate (DEHP), the most widely used phthalate plasticizer, has been implicated in skin cancer. However, its key targets and multi-pathway mechanisms regulating skin cancer onset and progression remain unclear. Therefore, elucidating DEHP's molecular mechanisms in skin cancer development is crucial for prevention and intervention strategies.<h4>Methods</h4>This study integrates network toxicology, molecular docking, and experimental validation to systematically elucidate the mechanism by which DEHP induces skin cancer. Specifically, we predicted potential DEHP targets and skin cancer-associated targets (melanoma, squamous cell carcinoma, basal cell carcinoma) using multiple databases. Core targets were identified through CytoNCA topological analysis, MCODE module mining, and CytoHubba multi-algorithm integration. Performed GO/KEGG enrichment analysis using the DAVID database; validated the binding potential between DEHP and core targets via molecular docking with AutoDock Vina; and finally verified the abnormal expression profiles of core targets through TCGA/GTEx transcriptomic data, HPA proteomic data, and RT-qPCR experiments in A-375 (human malignant melanoma cells)/Hacat cells.<h4>Results</h4>Initially, this study identified 11 key DEHP-induced skin cancer targets: CTNNB1, ESR1, HIF1A, IL6, MTOR, MYC, STAT3, AKT1, BCL2, CASP3, and CCND1. These targets exhibited specific regulation across different skin cancer subtypes. Subsequently, a four-tier regulatory network linking "DEHP-core targets-pathways-skin cancer" was constructed. Molecular docking confirmed stable binding conformations between DEHP and all 11 key targets, while enrichment analysis revealed their associations with cellular proliferation, apoptosis, inflammatory responses, and core pathways including Jak-STAT and PI3K-Akt/mTOR. Finally, transcriptomic, proteomic, and <i>in vitro</i> cellular experiments validated the significant dysregulation of these targets in skin cancer tissues and tumor cells.<h4>Conclusion</h4>Collectively, this study systematically elucidates the toxicological mechanism by which DEHP promotes skin cancer development through subtype-specific pathways regulated by 11 key targets, clarifying its direct binding patterns with core targets and downstream pathway disruption characteristics. This not only fills a research gap in the molecular mechanisms of DEHP-induced skin carcinogenesis but also provides novel biomarkers for environmental exposure prevention and targeted interventions against skin cancer.

Empty sella syndrome (ESS) is a radiological condition characterized by herniation of cerebrospinal fluid into the sella turcica, resulting in compression of the pituitary gland. Although often incidental, ESS may be associated with selective pituitary hormone deficiencies, including gonadotropin deficiency, leading to hypogonadotropic hypogonadism (HH) and infertility. We report the case of a 34-year-old woman with 14 years of primary infertility and a history of primary amenorrhea. Hormonal assessment revealed profound gonadotropin deficiency with a follicle-stimulating hormone (FSH) of 0.39 IU/L, luteinizing hormone (LH) less than 0.10 IU/L, and estradiol below 5 pg/mL, with preserved thyroid and prolactin function. Anti-Müllerian hormone (AMH) level was 0.8 ng/mL. Pituitary magnetic resonance imaging (MRI) demonstrated a partial empty sella without adenoma. Hysterosalpingography and semen analysis were normal. Ovulation induction with human menopausal gonadotropin resulted in pregnancy after one stimulation cycle. Cesarean section was performed at 38+3 weeks of gestation, delivering a healthy male newborn weighing 3180 g with an Apgar score of 10/10. Partial empty sella syndrome may be associated with isolated central hypogonadism leading to infertility, and individualized gonadotropin therapy can restore ovulation and achieve a successful pregnancy outcome.

<title>Abstract</title> <p>Purpose Clinically useful glioblastoma (GBM)-specific stemness biomarker programs remain incompletely defined. We aimed to prioritize a GBM-focused stemness program and candidate markers by integrating single-cell, single-nucleus, and bulk transcriptomic data. Methods We integrated scRNA-seq data from 55 GBM samples and snRNA-seq data from 10 GBM samples. Malignant-cell stemness was quantified with CytoTRACE. Stem.Sig was derived from six public GBM single-cell datasets by intersecting genes positively associated with CytoTRACE with genes upregulated in malignant cells. Bulk cohorts were examined for immune associations, tumor-type summary trends with intratumoral heterogeneity (ITH) and tumor mutational burden (TMB), and small-cohort anti-PD-1 outcome modeling. Weighted gene co-expression network analysis (WGCNA) was used to prioritize hub genes, followed by preliminary qPCR and western blot expression support. Results Malignant compartments associated with non-response showed higher CytoTRACE scores than treatment-naive compartments in both cellular and nuclear datasets (both P < 0.001). Stem.Sig was associated with lower immune-related gene expression and lower inferred immune-cell abundance. At the tumor-type summary level, higher median Stem.Sig tended to align with higher ITH and TMB; these comparisons were interpreted descriptively. A Naive Bayes exploratory model showed modest discrimination in pretreatment anti-PD-1 GBM cohorts (validation and test AUCs both 0.65). WGCNA overlap prioritized nine hub genes; SOX6, BCHE, and LINC00511 showed preliminary qPCR support, and SOX6 and BCHE were also supported at the protein level. Conclusion This integrative framework prioritizes a GBM stemness program linked to immune-cold biology, heterogeneity, and adverse outcome, and nominates SOX6, BCHE, and LINC00511 as candidate markers for mechanistic follow-up.</p>

While the majority of rodent resuscitation studies prioritize neurologic outcomes, research into the long-term effects of global ischemia and reperfusion on cardiac function is scarce, generally limited to morphologic assessments. Extracorporeal cardiopulmonary resuscitation (ECPR) is a promising strategy for highly selected patients with refractory cardiac arrest (CA). We aimed to investigate the impact of ventricular fibrillation CA (VFCA) and subsequent ECPR on cardiac recovery in rats. Adult male Sprague-Dawley rats were subjected to either 6-8 min VFCA, followed by ECPR and compared to sham animals. The primary outcome parameter was cardiac function assessment at 14 days after CA. The hearts of rats surviving for 14 days were isolated and mounted onto an erythrocyte-perfused, isolated working heart (WH) system. Cardiac output (CO), left ventricular systolic pressure (LVSP), and coronary flow were measured. To assess the heart adaptation to hemodynamic stress, the afterload was gradually increased in 10 mmHg increments while CO and LVSP were monitored. Additionally, the hearts from all animals surviving at least 36 h were assessed histologically. Of 15 rats that achieved ROSC after 6 min of CA, 7 could be evaluated in the WH setup 14 days after CA. In the 8 min CA group, 15 animals achieved ROSC, of which 2 were investigated in WH at 14 days after CA. Compared to the hearts of 7 sham animals, no significant differences in cardiac hemodynamics were observed at a set afterload (60 mm Hg; baseline) in the 6 min CA group. However, the two investigated 8 min CA animals exhibited a trend towards reduced CO and LVSP levels. Notably, both CA groups showed impaired hemodynamic performance to hemodynamic stress. Survivors at 14 days consistently showed significant myocardial pathology, with both 6 min and 8 min CA groups exhibiting fibrosis, inflammation, and edema most pronounced in the interventricular septum and right ventricle of the 8 min CA group. Animals that died prematurely displayed time-dependent acute changes, progressing from hypereosinophilic degeneration (36 h survivors) to myocardial necrosis, calcification, and the formation of cell-rich granulation tissue (48-108 h survivors). VFCA led to impaired left ventricular hemodynamic function in 8 min CA rats resuscitated with ECPR at rest and with increasing afterload. The isolated WH system may offer a valuable tool for assessing long-term cardiac function and performance after resuscitation.

Binaphthoquinone (BiNQ), a dimeric compound composed of juglone (5-hydroxy-1,4-naphthoquinone) and naphthazarin (5,8-dihydroxy-1,4-naphthoquinone), was originally isolated from the sea urchin <i>Spatangus purpureus</i> O.F. Müller. In this study, we examined the anti-fibrotic effects of BiNQ in a diabetic context, focusing on its potential to enhance mitochondrial function. Human cardiac fibroblasts (NHCF-V) were cultured under control 5 mM glucose or exposed to 33 mM glucose plus palmitic acid (100 μM). Fibroblast-to-myofibroblast transition was evaluated by measuring the expression level of the myofibroblast marker α-smooth muscle actin (α-SMA), and functional changes were further assessed using a contraction assay. BiNQ was applied at two concentrations (10 nM and 1 μM) and was found to reduce α-SMA expression, indicating inhibition of myofibroblast differentiation. Concurrently, BiNQ enhanced the expression of mitochondrial respiratory chain complexes, resulting in increased ATP production. To elucidate the underlying molecular mechanisms, targeted proteomic analysis was performed using LC-MS/MS, enabling identification and quantification of proteins involved in mitochondrial bioenergetics and fibrotic regulation. Collectively, these findings suggest that BiNQ attenuates fibroblast activation in a diabetic context by improving mitochondrial function and modulating key pro-fibrotic pathways.

Mechanochemical methodologies are reshaping synthetic organic chemistry by enhancing practicality and reducing environmental impact. This review presents a comprehensive account of mechanochemical methods for amide bond formation, arguably the most developed and industrially relevant area within mechanochemical organic synthesis. Covering literature from early contributions to the present (September 2025), the review organization follows key substrate classes and methodological strategies: amide bond formation via coupling of carboxylic acids or their activated derivatives with amines (Section 2), followed by unconventional approaches (Section 3) employing carboxylic acid and amine surrogates, redox chemistry, rearrangements, and transition metal-mediated reactions leading to amide products through alternative bond-forming pathways. Mechanoenzymatic transformations are treated separately (Section 4), with stereochemistry-related issues, such as the preservation of enantiomeric purity, discussed in Section 5. Section 6 highlights the use of amide bond formation as a model system for probing mechanochemical driving forces. Special attention is given to scalability and successful scale-up examples, alignment with green chemistry principles, limitations, unexplored areas, and challenges requiring further development. This review is intended as an accessible and thorough resource for synthetic chemists in both academia and industry, including those newly exploring the field of mechanochemistry, and it provides practical guidance for process optimization and scale-up.

<h4>Background</h4>Colorectal cancer (CRC), a lethal tumor of the intestine, has lipotoxicity, which exerts a significant influence on the onset and development of intestinal disorders. This study provides an in-depth evaluation of lipotoxicity-related genes (LRGs) in the CRC treatment setting.<h4>Methods</h4>Differentially expressed genes (DEGs) within The Cancer Genome Atlas-CRC dataset and LRGs were intersected to obtain the LRG-DEGs. Subsequently, univariate Cox regression and a random survival forest model were conducted to select prognostic LRGs and construct a risk model for patients with CRC. Then, prognosis was further investigated via independent prognostic analysis, nomogram construction, enrichment analysis, immune microenvironment analysis, and drug sensitivity testing. Finally, reverse transcription quantitative PCR (RT-qPCR) was used to verify the expression levels of prognostic LRGs.<h4>Results</h4>The constructed risk model with five prognostic LRGs (PPARGC1A, CPT2, CXCL1, FABP4, and OFCC1) showed high prognostic effectiveness. The nomogram subsequently built based on risk score, age, T stage, and M stage showed strong prognostic power. Moreover, the enrichment of diverse pathways was observed across distinct risk groups, exemplified by the PPAR signaling pathway and complement and coagulation cascades. Analysis of the immune microenvironment revealed the strongest positive association between FABP4 and natural killer cells. Drug sensitivity testing identified efficacious drugs for patients with CRC, such as midostaurin and lenalidomide. Notably, RT-qPCR confirmed elevated expression levels for CXCL1 and OFCC1 and reduced levels for FABP4, PPARGC1A, and CPT2 in patients with CRC.<h4>Conclusion</h4>Five prognostic LRGs were determined for CRC, and a new risk model was developed and validated, revealing the critical role of LRGs in CRC and improving our understanding of clinical interventions for this cancer type.

The dysregulation of proteostasis is a hallmark of Alzheimer's disease (AD), characterized by the accumulation of misfolded and aggregated proteins. Dysfunction of the ubiquitin-proteasome pathway is a major contributing factor to proteostasis imbalance. The E3 ubiquitin ligase, F-box and WD repeat domain-containing 7 (FBXW7), a key hub factor in AD, is significantly downregulated in AD patients. FBXW7 mediates the proteasomal degradation of tau and regulates the development of tau pathology. However, the effect of FBXW7 on β-amyloid pathology and the underlying mechanisms remain unclear. This study demonstrated that FBXW7α, the dominant FBXW7 isoform, was localized in both the cytoplasm and nucleus of neurons. Aging led to a decline in FBXW7α protein levels in the brain tissues of both wild-type and 5×FAD mice. Notably, the level of FBXW7 in the brain tissue of 5×FAD mice is significantly lower than that in wild-type mice after 6 months of age. FBXW7α interacted with BACE1 via the conserved phosphodegron motif and targeted BACE1 for degradation. FBXW7 knockdown diminished the ubiquitination of BACE1, impaired its proteasome-mediated degradation, and increased the accumulation of BACE1 in Golgi fractions. Additionally, restoration of FBXW7α in the hippocampus improved cognitive function and ameliorated amyloid pathology in 5×FAD mice. Our findings suggest that FBXW7α acts as a key regulator of amyloid pathology, and highlight FBXW7α as a promising potential therapeutic target for AD intervention.

Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterized by cognitive decline and synaptic dysfunction. Among the earliest regions affected is the retrosplenial cortex (RSC), where parvalbumin-expressing (PV + ) interneurons are particularly susceptible to AD-related pathology. To understand the molecular alterations within these vulnerable neurons we employed a dual-platform spatial transcriptomics approach, integrating GeoMx Digital Spatial Profiler (DSP) and Xenium In Situ. We analyzed the transcriptomic profiles of PV+ and NeuN+ neurons in the RSC of female 5xFAD mice. We leveraged the individual strengths of each platform to generate a robust and comprehensive dataset. Using non-negative matrix factorization and k-means clustering, we identified disease-associated metagenes and examined their spatial distribution. Our analysis revealed distinct transcriptional subpopulations within PV+ interneurons, with specific metagenes differentially expressed in RSC. Dner, Gad1, and Pvalb exhibited significant down-regulation in TG mice, suggesting impairments in PV+ interneuron function and GABAergic signalling. Cross-validation between GeoMx DSP and Xenium In Situ as well as RNAscope and immunohistochemistry confirmed the reproducibility and robustness of these findings. This study provides insights into the heterogeneity and molecular vulnerabilities of PV+ interneurons in AD and demonstrates the power of integrating spatial transcriptomic platforms to uncover disease-associated neuronal subtypes and molecular markers.

Interactions between the brain and immune system play a key role in the aetiology of brain disorders, with inflammation emerging as a potential causal factor in subsets of major depressive disorder, particularly those resistant to treatment. The mechanisms through which immune activation might drive depressive symptoms remain elusive, limiting the ability to develop new targeted therapies. Using a mouse model of neuroinflammation, involving a TLR7/8 agonist, we combined whole-brain and spatial transcriptomics approaches to determine whether TLR7/8-driven neuroinflammation resulted in consistent immune and neurobiological transcriptional changes throughout the brain. We found evidence of strong immune activation throughout the brain,. and this global inflammatory signal led to regionally specific changes in gene expression. In particular, we found reduced expression of genes associated with synaptic function in brain areas underlying mood and anxiety, such as ventral striatum and amygdala.