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Elevated intratumoral immune inflammation prior to treatment is typically associated with better outcomes in hot tumors treated with immune checkpoint inhibitors (ICIs). However, we observed a paradox in pMMR/MSS locally advanced rectal cancer (LARC) patients, where a subset with elevated baseline immune inflammation exhibited worse outcomes after combined radiotherapy and ICI treatment compared with patients with minimal immune inflammation. To investigate this counterintuitive finding, we performed paired scRNA-seq and scTCR-seq on longitudinally collected samples, including tumor biopsies (pre-treatment, post-radiotherapy, and post-immunotherapy) and peripheral blood mononuclear cells (pre-treatment and post-immunotherapy), from 20 pMMR/MSS LARC patients treated with sequential radiotherapy and ICI therapy (NCT06493240). We propose the concept of clonal entrapment to explain this phenomenon. Specifically, our profiling results reveal that increased HLA-DQA2 expression in dendritic cells and upregulated GDF15 expression in treatment-resistant tumor cells correlate with the restricted expansion of novel tumor-reactive TCR clonotypes. Consequently, the immune response is limited primarily by pre-existing TCR clonotypes within the tumor, especially those partially expanded under chronic inflammation, leading to the expansion of TCR clonotypes derived mainly from pre-treatment CD8⁺ T cell pools following ICI therapy. By identifying this feature of the pMMR/MSS LARC microenvironment, our study provides a high-resolution framework for understanding resistance to sequential radiotherapy and ICI therapy.

Extrachromosomal DNA (ecDNA) drives extensive oncogene amplification and intrinsic tumor heterogeneity in human cancers. Recent studies have uncovered an unexpected convergence between viral oncogenesis and ecDNA biology: integration of human papillomavirus (HPV) can trigger the formation of hybrid viral-human ecDNA structures in approximately 30% of HPV-associated oropharyngeal cancers (HPVOPC), with analogous structures reported in cervical cancer models. These chimeric circular elements fuse viral oncogenes with captured human genomic sequences, co-amplifying viral and human genes, and generating <i>de novo</i> enhancer complexes absent from either parental structure. Hybrid ecDNA exhibits canonical ecDNA features, including disproportionately high gene expression, accessible chromatin landscape, and the capacity to act as mobile trans-activating elements that potentiate oncogenic programs. Emerging evidence further suggests that hybrid ecDNA operates through distinct regulatory mechanisms and may create specific therapeutic vulnerabilities that can be leveraged for personalized cancer therapy. Elucidation of the mechanisms governing hybrid ecDNA biogenesis, maintenance, and function may therefore uncover exploitable dependencies in hybrid ecDNA-associated cancers. In this review, we synthesize current insights into hybrid ecDNA formation, regulatory mechanisms, and therapeutic opportunities and discuss how these advances may guide the development of precision cancer therapies.

Tissue regeneration requires de novo patterning, which has been proposed to be facilitated by cellular heterogeneity. Yet how such heterogeneities are integrated with the mechanochemical state of the tissue and stabilized at the chromatin level into stable, spatially organized fates remains poorly understood. Using in vivo mouse intestinal regeneration models and organoids, we identify a critical density regime that produces a permissive window for heterogeneity in the mechanosensor Yes-associated protein 1 (YAP1). We show that YAP1 heterogeneity is coupled to lineage-biased chromatin accessibility and is decoded through FOXA1, which integrates the permissive chromatin state to Delta-Notch supracellular feedback and lineage commitment. This circuit generates fate bistability and preserves a memory of transient YAP1 activity, thereby maintaining spatial patterning as tissues return to homeostasis after injury. Together, our findings establish a multiscale framework in which tissue-scale mechanics tune single-cell competence and, through FOXA1-mediated bistability, convert transient heterogeneity into stable and self-organized tissue architecture.

<h4>Background</h4>Metabolic competition and nutrient restriction in the tumor microenvironment (TME) shape the immune infiltrate in tumors and subsequently tumor immunity. In this study we used the transgenic melanoma mouse model tg(Grm1)EPv, which spontaneously develops melanoma due to the ectopic expression of the metabotropic glutamate receptor 1 (<i>Grm1</i>) in melanocytes to investigate if aberrant glutamate metabolism drives tumor formation and affects immune cell function.<h4>Methods</h4>We performed liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based metabolomic analyses and RNA sequencing on tumor-free and tumor-bearing tg(Grm1)EPv tissues to characterize metabolic alterations associated with tumor progression. Flow cytometry was used to examine changes in immune cell subsets within the TME. To assess the functional relevance of glutamate metabolism, we inhibited glutathione metabolism using L-buthionine-(S,R)-sulfoximine (BSO), an inhibitor of glutamate-cysteine ligase that depletes cellular glutathione levels.<h4>Results</h4>LC-MS/MS-based metabolomic analyses and RNA sequencing revealed changes in glutamate and glutamine metabolism, a glycolytic shift (Warburg effect), and reduced ATP levels in advanced tumors compared with tumor-free tissue, suggesting respiratory chain dysfunction. These metabolic changes in the TME are advantageous for the tumor cells and unfavorable for immune cells, such as dendritic cells (DC). Indeed, flow cytometry analysis of myeloid subsets during tumor progression showed a decline in tumor-infiltrating conventional type 2 DC and macrophages, alongside an increase in neutrophil and monocyte populations in advanced lesions. Interference with glutamate metabolism using BSO induced immunogenic cell death, namely ferroptosis, in an tg(Grm1)EPv-derived cell line in vitro. Therefore, we evaluated the combination of this inhibitor with immunotherapy as a promising new approach for the treatment of tumors in the tg(Grm1)EPv mouse model. We observed that tumor growth could be delayed in vivo when BSO was combined with a therapy regimen boosting DC numbers and activation. This inhibition of tumor growth was supported by the infiltration of activated T cells.<h4>Conclusion</h4>Overall, our findings provide novel insights into the importance of combining metabolic intervention with immunotherapy for the treatment of patients with melanoma, particularly those bearing glutamate pathway-active or immunologically cold tumors. This knowledge can drive the design of novel therapeutic strategies for patients with cancer.

Life on Earth has evolved in a form suitable for the gravitational force. Although the pivotal role of gravity in gene expression has been suggested, the molecular details remain unclear. Here, we show that mitochondria utilize gravity to activate protein synthesis within the organelle. Genome-wide ribosome profiling reveals reduced mitochondrial translation in mammalian cells and Caenorhabditis elegans under microgravity. We found that attenuation of cell adhesion through laminin-integrin interactions caused the phenotype. Mitochondrial translation is activated by a signal relayed by FAK, RAC1, PAK1, BAD, and Bcl-2 family proteins in the cytosol, and the mitochondrial fatty acid synthesis (mtFAS) pathway in the matrix. Consumption of mitochondrial malonyl-CoA by mtFAS reduces the malonylation of the translational machinery and accelerates the rates of translational initiation and elongation. Physiologically, this system operates in mechano-response of skeletal muscles. Our work provides mechanistic insights into how cells convert gravitational and mechanical forces into translation in mitochondria.

We applied transcriptome tomography to create a whole-brain model of early-stage Huntington's disease (HD) in R6/2 mice, which ubiquitously express truncated human mutant HTT containing approximately 150 CAG repeats. Medium spiny neuron (MSN)-related genes showed abnormal expression in the HD brain, in terms of expression similarity to wild-type Htt. Bdnf was the most probable upstream regulator of these genes. Smarca4, the Bdnf-regulator, was similarly expressed to wild-type Htt in the control brain; however, this was not observed in HD, implying a possible involvement of Smarca4 in glutamate excitotoxicity in HD. Lhx6, a master gene for MSN-related pathway development ordinarily conserved postnatally and in adulthood, was lost in the HD co-expression network. And a network hub node, Fcho1, was lost in connection involving Lhx6 and mitochondrial gene clusters. Loss of Smarca4, Lhx6, and Fcho1 in co-expression may contribute to spatiotemporally specific neuronal loss in HD.

Three-dimensional genomics methods such as Hi-C and Micro-C have uncovered chromatin loops across the genome and linked these loops to gene regulation. However, these methods only measure three-dimensional interaction probabilities on a relative scale. Here we overcome this limitation by using live-imaging data to calibrate Micro-C in mouse embryonic stem cells, thus obtaining absolute looping probabilities for 65,929 Micro-C-identified chromatin loops. We find that the looped state is generally rare, with a mean pairwise looping probability of 1.2% and a maximum of 25% across the quantified loops. On average, CTCF-CTCF loops are stronger than cis-regulatory loops (2.2% versus <1%). Our findings can be extended to human cells with available Micro-C data under certain assumptions. Overall, we establish an approach for genome-wide absolute loop quantification and report that loops occur with low probabilities, generalizing recent live-imaging results to the whole genome.

National genome projects (NGPs) are increasingly shaping precision medicine by improving representation of population-specific genetic diversity. This review compiles findings from NGPs across Asia and Africa, regions that remain underrepresented in global genomic databases despite their extensive demographic and genetic diversity. A total of 53 studies from 24 countries were identified to understand (1) the genomic approach utilized, (2) novel findings that have emerged, and (3) strategies for improving research in these regions. The NGPs implement population-based variome databases (20 NGPs), linear reference genome assemblies (8 NGPs), and graph-based pangenome assemblies (1 NGP). Novel variants ranged between 0.28% (China) and 19.6% (Iran), whereas rare variants accounted for up to 88.9% of the detected variants in the Chinese population. Each NGP documents its country's evolutionary and migration history, which impacts disease frequency and pharmacogenomic variants. Clinically, NGPs revealed strong population stratification in disease-associated and pharmacogenomic variants. For example, the GJB2 rs72474224 hearing-loss variant ranged from 13% in Vietnam and 12% in Hong Kong to 0.0894% in Turkey, while the VKORC1 rs9923231 pharmacogenomic variant reached 89.2% in Taiwan but was 20%-25% in European-related Russian subpopulations. These findings demonstrate that clinically relevant allele frequencies, pathogenicity assessments, and drug-response markers differ substantially across ancestries. This review highlights ongoing efforts and strategies to enhance the representativeness of genomic data through NGPs in Asia and Africa. We also suggest future directions for national projects, including integrating family-based studies, multi-omic data, and standardized pipelines to accelerate discovery and support the equitable implementation of precision medicine.

Congenital factor XII (FXII) deficiency is a rare disorder resulting from F12 gene mutations and its precise contribution to thrombotic or haemorrhagic risk has yet to be established. We identified the same mutation of the F12 gene across five pedigrees, yet the clinical manifestations varied markedly, ranging from asymptomatic to thrombotic and haemorrhagic phenotypes. Haematological assessments and deoxyribonucleic acid sequencing were conducted in five pedigrees. All five probands displayed prolonged activated partial thromboplastin time, markedly reduced FXII activity and an identical frameshift mutation in F12 (c.303_304delCA, p.H101Qfs*36), leading to the loss of functional domains. In Pedigree A, characterized by thrombotic clustering, thrombotic events were documented in 4 of 6 mutation carriers and 4 of 13 non-carriers. In Pedigree B, the proband experienced bleeding events, and her father died of intracerebral haemorrhage. Probands C and D remained asymptomatic. Proband E was diagnosed with essential thrombocythaemia and thrombosis. Across all pedigrees, haemorrhagic or thrombotic events could be attributed to additional contributing factors. The identified apolipoprotein E and butyrophilin subfamily 2 member A1 variants were consistently associated with thrombotic events in Pedigree A. This F12 variant does not appear to independently drive pathological coagulation phenotypes but may potentiate susceptibility to thrombosis in the presence of additional genetic or acquired risk factors.

Influenced by the internet and social media, a growing number of individuals are taking hepatotoxic amounts of prescription and nonprescription drugs leading to drug-induced liver injuries (DILIs). We present a case of fenbendazole-associated DILI with concomitant use of tauroursodeoxycholic acid as an intended hepatoprotective agent. While fenbendazole-associated DILI has been increasingly reported, this case is unique due to associated tauroursodeoxycholic acid use, and, to our knowledge, represents the first published report of this exposure combination. This case builds on the growing literature of DILI due to nonprescription drugs and highlights the importance of patient-centered counseling.

<h4>Background</h4>Epithelial barrier dysfunction plays a key role in asthma pathogenesis. Palmatine has been shown to reinforce epithelial integrity in colitis and ameliorate urticaria-like skin inflammation, suggesting potential barrier-protective effects.<h4>Purpose</h4>To investigate the barrier-protective role of palmatine in eosinophilic asthma.<h4>Study design and methods</h4>An ovalbumin-induced eosinophilic asthma model was established. After palmatine administration, lung function, airway responsiveness, mucus secretion, collagen deposition, and epithelial barrier integrity were assessed. Proteomics, molecular docking, cellular experiments, and immunofluorescence were performed to elucidate underlying mechanisms.<h4>Results</h4>Palmatine markedly restored epithelial barrier integrity by upregulating E-cadherin, occludin, and ZO-1. Correspondingly, it improved both large- and small-airway obstruction, and reduced airway hyperresponsiveness. It also reduced IL-5, IL-13, and MMP9 levels, indicating attenuation of inflammation and collagen deposition. Proteomics and mechanistic studies revealed that palmatine suppressed Alox15-mediated ferroptosis in airway epithelial cells, reflected by reduced GSSG, MDA, and LPO and increased GSH levels in lung tissue. In erastin-treated BEAS-2B cells, palmatine inhibited Alox15 and PEBP1 expression accompanied by recovery of epithelial junction proteins E-cadherin, occludin, and ZO-1.<h4>Conclusion</h4>Palmatine exhibits therapeutic effects in eosinophilic asthma. By inhibiting Alox15-mediated ferroptosis, it restores airway epithelial barrier integrity and alleviates inflammation and airway hyperresponsiveness, highlighting its promise as a potential therapeutic agent for eosinophilic asthma.

<h4>Background</h4>Metabolic dysfunction-associated steatotic liver disease and its more severe manifestation, metabolic dysfunction-associated steatohepatitis (MASH), are intimately linked to genetic factors, gut microbiota and barrier alteration. Ceramidases and ceramides are associated with MASH, yet the role of intestinal neutral ceramidase in MASH development remains unclear.<h4>Methods</h4>Murine models with intestinal epithelial cell (IEC)-specific depletion of neutral ceramidase (<i>Asah2^ΔIEC</i> ) or aryl hydrocarbon receptor (<i>AhR</i> ^ΔIEC) were subjected to either a Western diet (WD) at 6 weeks old for 10-12 months to induce MASH, or a hydrogenated vegetable oil, sucrose, palmitate and cholesterol (HSPC) diet to accelerate MASH progression. Fecal microbiota transplantation was performed in germ-free mice.<h4>Results</h4>MASH is associated with the induction of neutral ceramidase, which reshapes the intestinal microbiota and metabolite profiles, leading to increased production of 2-hydroxyhippuric acid (2-HHA). We identified 2-HHA as an inhibitor of AhR signalling, a pathway that normally promotes intestinal fucosylation. Elevated 2-HHA suppresses AhR activity, reduces fucosylation and contributes to MASH and associated airway inflammation in mice fed WD or HSPC diet. Notably, IEC-specific deletion of neutral ceramidase decreases 2-HHA levels, restores AhR signalling, enhances fucosylation and protects against MASH. Consistently, intestinal AhR deficiency exacerbates MASH by reducing intestinal fucosylation, whereas supplementation with fucoidan increases fucosylation, improves barrier function and attenuates MASH.<h4>Conclusions</h4>These findings identify intestinal neutral ceramidase as a key driver of MASH through a microbiota-2-HHA-AhR axis that impairs intestinal fucosylation and barrier function, highlighting a potential therapeutic target.

<h4>Objective</h4>Chronic rhinosinusitis with nasal polyps (CRSwNP) is a heterogeneous chronic inflammatory disease with unclear pathogenesis and a high recurrence rate. Inflammatory response-related genes (IRGs) play a key role in disease progression but remain to be further explored in CRSwNP.<h4>Methods</h4>CRSwNP-related transcriptome datasets and single-cell RNA sequencing (scRNA-seq) data were obtained from the Gene Expression Omnibus (GEO) database. IRGs were derived from the Molecular Signatures Database (MSigDB). Core genes were screened using weighted gene co-expression network analysis (WGCNA) and protein-protein interaction (PPI) network analysis, and the expression of the core genes was experimentally validated. The CIBERSORT algorithm was applied to analyze the correlation between core genes and immune cell infiltration. scRNA-seq data were used to explore the expression patterns of core genes in different cell subsets. Finally, Connectivity Map (CMap) and molecular docking were combined to predict potential therapeutic drugs for CRSwNP.<h4>Results</h4>A total of 96 key genes were identified through differential expression analysis and WGCNA, which were mainly enriched in the IL-17 signaling pathway, chemokine-cytokine interaction, and cytokine-cytokine receptor interaction pathways. Among them, FOXP3, C5AR1 and LIF can serve as effective prognostic predictors for CRSwNP. CIBERSORT analysis showed that activated CD4⁺ memory T cells and M2-type macrophages were significantly enriched in CRSwNP tissues. CMap analysis predicted that bicuculline had potential therapeutic value for CRSwNP.<h4>Conclusion</h4>FOXP3, C5AR1 and LIF could serve as diagnostic biomarkers for CRSwNP and may participate in the pathogenesis of CRSwNP by regulating inflammatory responses and immune cell infiltration, which provides novel insights into clarifying the etiology and pathogenesis of CRSwNP.

<h4>Background</h4>Hypomethylating agents, particularly azacitidine and decitabine, are widely used for the treatment of myelodysplastic syndromes and acute myeloid leukemia. Gastrointestinal adverse events and metabolism- and nutrition-related disorders may compromise nutritional intake, metabolic homeostasis, treatment tolerance, and clinical outcomes. However, real-world evidence regarding these safety profiles remains limited.<h4>Objective</h4>This study aimed to characterize gastrointestinal and metabolism- and nutrition-related adverse event reporting signals associated with azacitidine and decitabine, examine cross-database signal consistency, assess exploratory time-to-onset patterns, and explore potential mechanistic clues.<h4>Methods</h4>FAERS reports from database inception through 2025 Q4 were analyzed for azacitidine and decitabine recorded as suspected drugs. Disproportionality was assessed at the preferred-term level using reporting odds ratio, proportional reporting ratio, Bayesian confidence propagation neural network, and multi-item gamma Poisson shrinker methods. Selected signals were examined in the Canada Vigilance database for cross-database signal consistency. Exploratory time-to-onset patterns were evaluated using Weibull modeling among reports with valid date information, and network toxicology was conducted as a hypothesis-generating analysis.<h4>Results</h4>Fourteen thousand two hundred sixteen azacitidine-related reports and 3,591 decitabine-related reports were included. Neutropenic colitis showed the strongest gastrointestinal disproportionality signal for azacitidine and decitabine, with reporting odds ratios of 23.65 and 26.05, respectively. Tumor lysis syndrome was the most prominent metabolism- and nutrition-related signal, with 165 azacitidine-related reports and 47 decitabine-related reports. Both signals showed consistent disproportional reporting patterns in the Canada Vigilance database. Additional signals included iron overload, hypoalbuminemia, electrolyte disturbances, cachexia, failure to thrive, and decreased appetite. Time-to-onset analysis suggested an early reporting tendency among reports with valid date information. Exploratory network toxicology identified 45 common targets, including <i>MMP9, PTGS2, CASP3, GSK3B</i>, and <i>ADAM17</i>, suggesting potential involvement of inflammation, apoptosis, matrix remodeling, nitrogen metabolism, folate biosynthesis, lipid metabolism, and insulin resistance pathways.<h4>Conclusion</h4>Azacitidine and decitabine showed clinically relevant gastrointestinal, metabolic, and nutrition-related disproportionality reporting signals in spontaneous reporting databases. Early monitoring of gastrointestinal symptoms, electrolyte balance, tumor lysis indicators, albumin levels, appetite, and iron metabolism may support nutritional risk management and individualized supportive care in patients receiving hypomethylating agents.

<h4>Background</h4>WD-repeat domain 1 (WDR1), a key regulator of actin cytoskeleton dynamics, has been implicated in tumor progression. However, its pan-cancer characteristics and immunological relevance remain unclear. In this study, we performed an integrative pan-cancer analysis to evaluate the expression patterns, prognostic value, molecular features, and immune associations of WDR1.<h4>Methods</h4>We systematically analyzed WDR1 across cancers using integrated datasets from The Cancer Genome Atlas (TCGA), Genotype-Tissue Expression (GTEx), and Gene Expression Omnibus (GEO). By integrating R software and online bioinformatics platforms (e.g., UALCAN, HPA, GEPIA2, cBioPortal, and TIMER2.0), we characterized WDR1 in terms of expression, prognostic and diagnostic value, epigenetic regulation, genetic variation, and immune cell infiltration. Functional enrichment and drug sensitivity analyses were further conducted, and <i>in vitro</i> experiments were performed for validation.<h4>Results</h4>WDR1 exhibited heterogeneous expression across cancers and showed tumor-type-dependent prognostic significance. In addition, WDR1 displayed differential DNA methylation patterns and was positively correlated with multiple RNA methylation-related regulators. Notably, WDR1 expression was closely associated with tumor immune microenvironment features, including increased infiltration of myeloid and stromal cells and broad correlations with immune checkpoint molecules and immune-related genes. Functional analyses indicated that WDR1-related genes were primarily involved in actin cytoskeleton organization. Drug sensitivity analysis revealed that WDR1 expression was associated with differential responses to multiple therapeutic agents. Importantly, experimental validation demonstrated that WDR1 exerted context-dependent biological effects, suppressing proliferation, migration, and invasion in renal cancer cells while promoting these phenotypes in gastric cancer cells.<h4>Conclusion</h4>Our findings demonstrated that WDR1 was associated with tumor progression and immune microenvironment remodeling in a context-dependent manner and may represent a potential biomarker for further investigation in cancer.

Camelliasaponins, bioactive constituents abundant in the by-products of <i>Camellia oleifera</i> oil production, exhibit diverse biological activities. However, their potential in regulating neuroprotective mitophagy remains largely unexplored. This study identifies camelliasaponin B1 (CSB1) as an abundant component in <i>C. oleifera</i> seeds and investigates its cytoprotective mechanisms against oxidative stress. Using an in vitro model of H₂O₂-induced oxidative damage in PC12 cells, we found that CSB1 pretreatment significantly alleviated oxidative stress, as evidenced by reduced reactive oxygen species (ROS) accumulation and enhanced antioxidant enzyme activities (SOD, CAT, GSH-Px). CSB1 also preserved mitochondrial function, restoring membrane potential (ΔΨm), ultrastructure, and respiratory capacity. Mechanistically, CSB1 reduces the expression of BNIP3/NIX-LC3B pathway-related proteins, suggesting a modulatory effect on mitophagy, as supported by transcriptomic analysis, Western blotting, and immunofluorescence. Molecular docking computationally predicted potential interactions between CSB1 and BNIP3/NIX proteins, which require experimental validation. Collectively, these findings suggest that CSB1 acts as a cytoprotective agent that enhances antioxidant defenses, safeguards mitochondrial integrity, and is associated with reduced BNIP3/NIX-LC3B expression and co-localization, offering a potential molecular basis for its development as a neuroprotective agent targeting oxidative stress-related mitochondrial dysfunction.

<h4>Background/objectives</h4>Hepatocellular carcinoma (HCC) arises through heterogeneous pathways involving chronic hepatitis B virus infection, hepatitis C virus infection, alcohol-related liver disease, metabolic dysfunction-associated steatotic liver disease, fibrosis, cirrhosis, and environmental exposures. Genome-wide association studies (GWASs) have identified host germline loci associated with HCC susceptibility, but interpretation is complicated by aetiology, ancestry, liver disease stage, and the definition of controls. This narrative review examines current GWAS evidence for HCC, with emphasis on aetiology-specific susceptibility, functional interpretation, cross-disorder genetic effects, and clinical translation.<h4>Methods</h4>Studies were identified through iterative searches of PubMed/PMC, publisher pages, academic search tools, and citation tracking, supplemented by targeted searches for major HCC-associated loci. Sources were chosen based on relevance to GWAS discovery, replication, meta-analysis, functional interpretation, polygenic risk modelling, or HCC risk stratification, rather than by a formal systematic review protocol.<h4>Results</h4>Viral HCC studies most often implicate immune regulation and antigen presentation, including <i>MICA</i>, <i>HLA-DQ</i>, <i>HLA-DQB1</i>, HLA class I, <i>HCP5</i>, <i>STAT4</i>, <i>DEPDC5</i>, and <i>FAM114A1</i>. Alcohol-related, metabolic, and non-viral HCC studies more often implicate hepatic lipid metabolism, telomere biology, iron metabolism, steatosis, and cirrhosis-related pathways, including <i>PNPLA3</i>, <i>TM6SF2</i>, <i>TERT</i>, <i>HSD17B13</i>, <i>APOE</i>, <i>HFE</i>, and <i>MTARC1</i>. Recent studies increasingly combine GWASs with fine-mapping, functional annotation, transcriptomic analyses, and risk modelling.<h4>Conclusions</h4>HCC genetic susceptibility is highly aetiology-specific and overlaps with other liver and metabolic disorders, but discoveries from genetic studies have not yet been translated into routine clinical practice. Future work should prioritise multi-ancestry cohorts, disease-stage-aware controls, functional validation, and prospectively tested genetic risk models.

<b>Background</b>: Primary ciliary dyskinesia (PCD) is a rare and genetically heterogeneous disorder that remains underdiagnosed in low- and middle-income countries, largely due to limited access to specialized diagnostic tests. Genetic analysis has become an essential component of PCD diagnosis, particularly where functional and ultrastructural evaluations are unavailable. <b>Methods</b>: We conducted an investigational study including children and adolescents with clinical suspicion of PCD followed at a Brazilian tertiary center. Clinical characterization included detailed phenotyping and calculation of the PICADAR score. Molecular investigation was performed using exome sequencing as a frontline diagnostic approach. <b>Results</b>: Among 27 individuals evaluated, 10 (37%) received a confirmed molecular diagnosis of PCD. An additional 6 (22%) individuals had inconclusive molecular findings, mainly due to variants of uncertain significance (VUS), and were classified as likely PCD based on combined clinical and molecular evidence. Higher PICADAR scores were more frequently observed among individuals with confirmed or likely molecular diagnosis, with 9 of 10 confirmed cases presenting a score above 5. Beyond PCD-associated findings, exome sequencing also enabled the identification of clinically relevant additional diagnoses, including cystic fibrosis, <i>FGFR3</i>-related hypochondroplasia, and ACMG-reportable secondary finding involving <i>BRCA2</i>. Some unresolved cases may also reflect inherent technical limitations of exome sequencing, including restricted sensitivity for copy-number variants, suboptimal coverage of highly homologous or GC-rich regions, and limited detection of deep intronic and other variants. Additional factors include challenges in variant interpretation and incomplete knowledge of disease-associated genes. <b>Conclusions</b>: Frontline exome sequencing is a valuable diagnostic tool for PCD, particularly when integrated with robust clinical phenotyping. Clinical scoring systems such as PICADAR may help prioritize individuals for genetic testing and optimize diagnostic yield in resource-limited settings.

<b>Background/Objectives</b>: Patient-derived cell lines retaining donor-specific age-related and genomic features are essential for modeling late-onset neurodegenerative disorders like Huntington's disease (HD). This study aims to establish and comprehensively characterize HDDF2, a novel dermal fibroblast line from an HD patient, to provide a relevant cellular model. <b>Methods</b>: Dermal fibroblasts were isolated and cultured from a 44-year-old male HD patient carrying 46 CAG repeats in the <i>HTT</i> gene. Cells were evaluated for senescence markers (p16, lamin B1, SA-β-Gal activity, proliferation rates) and polyglutamine (polyQ) aggregation. Direct reprogramming protocols were applied to convert these fibroblasts into induced neurons. <b>Results</b>: HDDF2 fibroblasts exhibited a pronounced senescence-associated phenotype, evidenced by increased p16 expression, reduced lamin B1 levels, elevated SA-β-Gal activity, and decreased proliferation. Notably, polyQ deposition was preferentially detected within the senescent subpopulation, displaying distinct localization patterns differentiating senescent from proliferating cells. Despite this, HDDF2 cells retained their capacity for direct reprogramming and were successfully converted into induced neurons. <b>Conclusions</b>: HDDF2 represents a well-characterized, patient-specific cellular model for HD. The observed co-occurrence of polyQ deposition and cellular senescence, combined with successful neuronal conversion, establishes this line as a valuable resource for investigating the relationship between cellular aging and HD pathogenesis.

Hepatocellular carcinoma (HCC) remains the third leading cause of cancer-related mortality worldwide, with limited treatment efficacy and poor prognosis, particularly in advanced-stage disease. Despite progress in diagnostic imaging and systemic therapies, early detection and effective targeted interventions remain major clinical challenges. Long non-coding RNAs (lncRNAs), a class of regulatory RNA molecules exceeding 200 nucleotides in length, have emerged as essential regulators of oncogenic signaling, metabolism, and tumor microenvironment in HCC. This review provides a comprehensive overview of the molecular mechanisms, biological functions, and clinical relevance of key lncRNAs involved in HCC progression. We summarize the dual roles of lncRNAs as oncogenic drivers and tumor suppressors, with particular emphasis on oncogenic lncRNAs including CYTOR, UCA1, MALAT1, SPRY4-IT1, uc001ncr, AF085935, HULC, and HOTAIR. Their regulatory functions are discussed within major cancer-associated signaling networks, including the PI3K/AKT/mTOR, Wnt/β-catenin, TGF-β/NF-κB, and EMT-associated pathways. Furthermore, each lncRNA is discussed in the context of its expression pattern, endogenous RNA (ceRNA) interactions, molecular partners, and functional contributions to cellular proliferation, invasion, metastasis, angiogenesis, immune modulation, and therapeutic resistance. In addition, we explore the emerging potential of lncRNAs as minimally invasive diagnostic and prognostic biomarkers, owing to their stability and detectability in tissue and circulating biofluids. We also discuss recent advances in RNA- targeted therapeutic strategies, including RNA interference, siRNA-mediated silencing, antisense oligonucleotides, and CRISPR-based genome editing approaches. Collectively, this review highlights the importance of integrating lncRNA expression signatures into personalized HCC management and the promising role of lncRNA-based diagnostics and therapeutics in transforming the future landscape of liver cancer detection, prognosis, and treatment.

The origin of the polybasic furin cleavage site (FCS) of SARS-CoV-2 remains a central question in debates on the emergence of COVID-19. One hypothesis proposes a genetic relationship between the SARS-CoV-2 S1/S2 motif RRAR and the RRVR sequence found in the mouse-adapted MERS-CoV strain MERS-MA30. Here, we combined large-scale bioinformatic analysis with experimental virology to evaluate this scenario. Analysis of over 17 million SARS-CoV-2 genomes revealed that the S:684V substitution corresponding to RRVR occurred repeatedly but only sporadically, never became phylogenetically basal, and showed limited geographic and temporal spread. Using reverse genetics, we generated SARS-CoV-2 variants encoding RRVR and demonstrated that S:684V consistently reduced viral entry efficiency and competitive fitness in multiple cell systems, including human respiratory epithelial cultures. RRVR variants did not evolve toward RRAR but instead accumulated alternative substitutions. These findings do not support an evolutionary relationship between MERS-MA30 and the SARS-CoV-2 FCS.

<h4>Background</h4>Natural mutations in the prothrombin gene, involving the substitution of Arg596 with polar/apolar amino acids, represent an important risk factor for thrombosis. Notably, Arg596 in prothrombin corresponds to position 221a in the Na⁺-binding loop of mature α-thrombin (αT). Patients carrying these mutations (Arg221a→Gln/Leu/Trp) share a common clinical phenotype, characterized by a moderately reduced procoagulant function in coagulation tests and resistance of αT variants to inactivation by antithrombin III (ATIII). However, the biochemical mechanisms linking these mutations to the increased thrombotic risk remain elusive.<h4>Objectives</h4>To provide quantitative data on the effect that the recombinant Arg221aTrp αT mutant (αT-PD2) has on the activation of procoagulant and anticoagulant pathways and elucidate the structural basis of the observed functional changes.<h4>Methods</h4>A multipronged approach was used, integrating recombinant DNA techniques, clotting/fibrinolytic and enzymatic assays, microscopy, proteomics, spectroscopic, mass spectrometry, and molecular modeling techniques.<h4>Results</h4>Arg221aTrp mutation abrogates physiological Na⁺ binding and reduces the proportion of the fully active αT conformation. αT-PD2 affects procoagulant pathways by less efficiently generating fibrin (7-fold) and activating platelets (10-fold). The mutation potently affects anticoagulant pathways, as αT-PD2 is less efficiently inhibited by ATIII (3-fold), binds heparin less tightly (10-fold), and much less efficiently (41-fold) activates protein C.<h4>Conclusion</h4>Our data indicate that the procoagulant phenotype, observed in Arg221aTrp mutation carriers, may result from opposing inhibitory effects on both procoagulant and anticoagulant pathways, with the impact on anticoagulant pathways being predominant and shifting the natural anticoagulant↔procoagulant equilibrium in vivo toward thrombosis.

While conventionally viewed as a defense against external pathogens, we posit that the adaptive immune system likely evolved primarily to preserve internal epithelial order. We propose that adaptive immunity, with combinatorial diversity emerging ~0.5 billion years ago during the Cambrian explosion, was based on extant and established complex cellular interactions, derived over the previous 3.5 billion years of life's history. These coordinate cellular processes originated to police epithelia against aberrant or rogue cells-inherent to complex multicellularity. Primordial surveillance relied on maintaining epithelial integrity and specialized immune sentinels. Early deposition of tissue-resident γδ T and natural killer cells detects abnormalities and emergent stress signals as damage-associated molecular pattern molecules (DAMPs), independently of classical major histocompatibility (MHC) presentation to αβ T cells. Trained immunity in tissue-resident macrophages, stroma, and epithelia reciprocally respond and are tailored by nascent and highly selected tissue-resident memory cells.

The switch from fetal to adult hemoglobin is tightly regulated during erythropoiesis, and its dysregulation can contribute to β-hemoglobinopathies. Although transcriptional repression of γ-globin by BCL11A is well established, the posttranscriptional mechanisms that sustain BCL11A expression in adult erythroid cells remain incompletely understood. Here, we report that the long noncoding RNA UCA1/miR-148b-mediated regulatory axis is critical for globin gene switching. We identified miR-148b as a direct posttranscriptional regulator of BCL11A. lncRNA UCA1, which is abundantly expressed in adult erythroid cells, functions as a molecular decoy for miR-148b, thereby attenuating miR-148b-mediated repression of BCL11A. Depletion of UCA1 increases miR-148b availability and reduces BCL11A expression, which can lead to robust induction of γ-globin. Conversely, the ectopic expression of UCA1 restores BCL11A levels by antagonizing miR-148b, thereby promoting γ-globin silencing in adult erythroid cells. Mechanistically, UCA1 orchestrates a posttranscriptional regulatory axis that reinforces γ-globin silencing by stabilizing BCL11A levels in adult erythroid cells. Taken together, our studies uncover a previously unrecognized lncRNA-mediated mechanism that integrates miRNA activity with transcriptional control to fine-tune hemoglobin switching during adult erythropoiesis.

The primary systemic treatment for advanced colorectal cancer (CRC) is chemotherapy, but inevitable drug resistance leads to poor prognosis. Cancer stem cells (CSCs) are recognized as significant contributors to chemotherapy resistance, yet the mechanisms underlying their ability to induce widespread resistance across tumor cell populations remain poorly understood. In this study, we demonstrate a novel mechanism by which colorectal CSC-like spheroid cells (CCSCs) drive chemotherapy resistance through the release of small extracellular vesicles (sEVs) enriched with highly phosphorylated insulin-like growth factor binding protein-3 (IGFBP3) into the tumor microenvironment (TME). While cell-free IGFBP3 or endogenous IGFBP3 directly induces apoptosis in tumor cells, IGFBP3 originating from quiescent CCSCs functions through an epidermal growth factor receptor (EGFR)-dependent pathway to promote chemotherapy resistance. Mechanistically, the protein kinase VRK2, which is highly enriched in CCSCs, leads to the phosphorylation of IGFBP3 at the S201 site. This phosphorylation enables IGFBP3 to bind to EGFR, facilitating its sorting and subsequent recruitment into sEVs. Both EGFR and phosphorylated IGFBP3 are co-delivered via sEVs to recipient tumor cells, where they inhibit chemotherapy-induced DNA damage and promote resistance. Importantly, siRNA targeting VRK2 or the inhibitor D4476 significantly reduced IGFBP3 phosphorylation and its incorporation into sEVs, thereby reversing chemotherapy resistance. These findings highlight the dual role of IGFBP3 in CRC and provide critical insights into the core mechanism by which CCSCs drive chemotherapy resistance through EGFR and sEVs-mediated signaling, offering a novel therapeutic approach for personalized treatment of CRC.

Huntington's disease research has focused on either loss of normal huntingtin function or toxic gain-of-function of the mutant huntingtin protein as a key driver of pathology. The role of mutant HTT mRNA in vivo has been only partially studied and remains largely unexplored. Recently, we discovered that full-length human HTT mRNA is retained, together with the alternatively processed HTT1a transcript, in RNA nuclear clusters in YAC128 mouse brains. Here, we demonstrate that these clusters were present at the prenatal stage, indicating early developmental effects. Moreover, these clusters were confined to neurons, implying a neuron-specific mechanism of accumulation, and were colocalised with core spliceosomal proteins, suggesting an impact on nuclear homeostasis. HTT nuclear RNA clusters showed remarkable dynamics, rapidly dissolving when ionic interactions were disrupted or transcription and splicing were inhibited. This malleability underscores the importance of HTT mRNA accessibility to therapeutic interventions and may guide future design of HTT-targeting therapies.

Low-grade serous carcinoma of the ovary (LGSOC) is relatively resistant to chemotherapy. Given its biological parallels to hormone receptor-positive breast cancer, including responsiveness to anti-estrogen therapies, we conducted a pilot phase II study to assess clinical benefit of neoadjuvant fulvestrant and abemaciclib in women with advanced, unresectable LGSOC (NCT03531645). Imaging assessments were performed every 8 weeks until resectable. The primary endpoint was clinical benefit rate (CBR). Exploratory objectives included evaluation of safety profile, accessing biomarkers related to clinical benefit, and description of tumor phenotypic changes. Fifteen patients were enrolled and evaluable for efficacy. CBR was 100%, with 1/15 patients (7%) achieving complete response (CR), 8/15 patients (53%) achieving partial response (PR), and 6/15 (40%) exhibiting stable disease (SD). Interval cytoreductive surgery (ICS) was performed in 9 patients (60%), with 7 (78%) achieving complete or optimal resection. Fourteen tumor samples underwent transcriptomic and proteomic profiling. Tumors from long-term survivors showed significantly higher baseline expression of cell-cycle-related programs and estrogen signaling-related genes, both suppressed upon treatment. Neoadjuvant fulvestrant and abemaciclib was well tolerated, achieved high response rates, and enabled optimal surgical resection in most patients. Tumor proliferative activity and estrogen signaling dependency may predict therapy response.

Heart failure (HF) remains a pressing health concern, with rising prevalence globally. Subjectivity and ambiguity in the definition of HF and its antecedent stages have limited research, global surveillance, and prevention programs. To address this, several cardiac societies and foundations convened to standardize the definition of HF in 2021 and designated stage B or pre-HF to identify individuals at risk of developing HF. In subsequent years, substantial progress and changes have been made in aspects of preventing HF, improving HF diagnosis and management, and recognizing the importance of the affected individual's voice. Global differences and disparities in HF are better understood, as are causes and comorbidities leading to differences in care, which are also influenced by access to care. This consensus document presents the Second Universal Definition of Heart Failure, aiming to standardize terminology and facilitate a uniform approach for clinicians, researchers, health systems, and policymakers. In this definition, the classification of HF phenotypes moves away from rigid left ventricular ejection fraction cutoffs, instead grouping HF into reduced, preserved, and improved ejection fraction categories to better reflect clinical realities. A universal classification of HF causes is also proposed. The document also addresses the dynamic trajectories of HF-improvement, remission, and recovery-and highlights the impact of social determinants and geographic variation on HF risk and outcomes. By providing a comprehensive, standardized framework for HF definition and classification, this document seeks to improve prevention, early detection, and management of HF worldwide, ultimately enhancing patient care and advancing global cardiovascular health.

Bone is a multifunctional organ essential for structural support, protection, hematopoiesis, and endocrine regulation. Emerging research increasingly highlights the pivotal role of the nervous system in bone metabolism, repair, and musculoskeletal disease progression, particularly the autonomic nervous system (ANS), which precisely regulates bone tissue cells via sympathetic and parasympathetic pathways. The ANS regulates the activity of mesenchymal stem cells (MSCs), osteoblasts, osteoclasts, chondrocytes, and nucleus pulposus cells (NPCs) via neurotransmitters such as norepinephrine (NE), acetylcholine (ACh), neuropeptide Y (NPY), and vasoactive intestinal peptide (VIP), thereby modulating core pathological processes in bone-related disorders including osteoporosis (OP), osteoarthritis (OA), intervertebral disc degeneration (IVDD), and traumatic fractures. Targeted neuromodulation is emerging as a promising therapeutic strategy for musculoskeletal disorders. This review aims to summarize the regulatory roles of the ANS in bone cell metabolism and bone-related pathologies and to evaluate current ANS-bone axis-based interventions to promote skeletal health and alleviate the burden of musculoskeletal diseases.

Ureteral stents are essential for relieving urinary obstruction and preserving renal function. However, long-term indwelling often induces friction-mediated urothelial irritation, bacterial colonization, and mineral encrustation, which impair urinary drainage and complicate stent removal. Here, we developed a friction-lowering urinary interface for defense against encrustation (FLUID) by integrating a silicone-based primer and lubricant layer into a fully microporous thermoplastic polyurethane framework. Unlike conventional slippery liquid-infused porous surfaces that retain lubricant primarily within surface microstructures, the fully microporous TPU framework serves as an internal lubricant reservoir throughout the stent wall, replenishing the lubricating interface under dynamic urinary conditions. Under sterile conditions the FLUID coating suppressed planktonic bacterial adhesion (<i>E. coli</i>, <i>B. cereus</i>, <i>S. aureus</i>; 72 h adhesion assay) by more than 94% relative to uncoated substrates. In a <i>P. mirabilis</i>-spiked 14-day mature biofilm model, biofilm-associated bacterial recovery from FLUID was modestly lower than from the commercialized comparators (Log R ≈ 0.10 and 0.14; approximately 21% and 28% reduction). In a porcine model, the coated stent reduced inflammatory activity and attenuated mineral accumulation after 4 weeks, with magnesium and calcium deposition decreased by 84.1% and 45.1%, respectively. After 8 weeks, scanning electron microscopy confirmed clean surfaces with patent side holes and minimal debris accumulation. Overall, the FLUID stent establishes a self-replenishing lubricant interface integrated throughout the microporous stent wall, enabling simultaneous mitigation of friction, early biofouling, and mineral deposition, and providing a clinically relevant proof-of-concept strategy for safer long-term urinary drainage that will require further preclinical validation prior to clinical adoption.

Cartilage has a limited intrinsic capacity for self-repair, causing injuries to progress into degenerative disorders. Current conservative and surgical treatments offer only partial benefits and are often hindered by poor tissue quality, limited durability, and procedural invasiveness. Functionalized microspheres have recently gained attention in cartilage repair due to their biocompatibility, injectability, structural versatility, and ability to achieve controlled therapeutic delivery. Evolving from passive carriers to smart, bioactive platforms, these microspheres can modulate the local regenerative microenvironment through programmable release, structural engineering, and responsive functionalities. This review summarizes the structure and degeneration of articular cartilage and the cartilaginous endplate, introduces major microsphere fabrication techniques and biomaterial choices, and highlights emerging strategies for functionalization. We further discuss structural innovations that enhance therapeutic outcomes and examine the translational potential of microspheres as precise and multifunctional smart microtissues for personalized cartilage regeneration.

This study developed an intelligent drug-loaded micelle system based on folic acid-conjugated polyglycidol (FAPG), which incorporates a reactive oxygen species (ROS) responsive fluorophore-drug conjugate (BTBP-PTX, BP). The system integrates folate receptor-α (FR-α) targeting, esterase/ROS dual-responsive drug release, and aggregation-induced emission (AIE)-based fluorescence activation for tumor-targeted delivery and imaging. Under simulated tumor microenvironment conditions (esterase-rich, high ROS levels), the system displayed dual-responsive release behavior, achieving over 80% paclitaxel (PTX) release within 24 h through esterase-catalyzed ester bond cleavage and ROS-triggered oxalate bond hydrolysis. Cellular assays demonstrated that BP@FAPG enhanced cellular uptake by approximately 2.5-fold in MDA-MB-231 cells compared to non-targeted controls BP@GAPG, along with significant cytotoxicity (IC₅₀ = 60 nM) and deep penetration into three-dimensional tumor spheroids-effects primarily attributed to FR-α-mediated endocytosis. Furthermore, the AIE fluorophore was activated during drug release, allowing real-time fluorescence tracking of micelle localization and release dynamics.

Human papillomavirus (HPV) represents one of the most clinically significant oncogenic viruses affecting human populations worldwide [...].

Pancreatic ductal adenocarcinoma is highly malignant with poor prognosis. Its dense tumor microenvironment severely limits the efficacy of conventional chemotherapy and causes severe side-effects. Herein, we adopt the established Schiff-base crosslinked thermoresponsive injectable self-healing poly(2-(2-methoxyethoxy)ethyl methacrylate-<i>co</i>-oligo(ethylene glycol) methyl ether methacrylate-<i>co</i>-aldehyde 2-hydroxyethyl methacrylate)/carboxymethyl chitosan (APMOH/CMCS) hydrogel as the delivery scaffold. By regulating monomer composition, the volume phase transition temperature (T_VPT) of the hydrogel was tuned to around 43 °C to match the therapeutic temperature requirement. Subsequently, copper-metal organic framework (Cu-MOF) nanoparticles co-loaded with 2,2'-azobis(2-methylimidazoline) dihydrochloride (AIPH) and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) cationic radicals (ABTS·⁺) (denoted as AB@Cu-MOF) were uniformly incorporated into the hydrogel network. Under near-infrared (NIR) irradiation, ABTS·⁺ acts as a photothermal agent to generate hyperthermia for tumor ablation; the elevated temperature further activates AIPH to produce alkyl radicals, which can oxidize inactivated ABTS back to ABTS·⁺ and construct a sustainable photothermal therapy-thermodynamic therapy (PTT-TDT) circulation. Meanwhile, Cu-MOF can consume intracellular glutathione (GSH) to protect active components from deactivation and initiate chemodynamic therapy (CDT) via Fenton-like reactions to produce toxic reactive oxygen species. Benefiting from the thermoresponsive characteristic, the hydrogel undergoes volume shrinkage upon heating, achieving NIR-triggered on-demand drug release with a cumulative release rate of 81.1%. In vitro and in vivo experiments verified that this integrated platform realizes remarkable triple synergistic efficacy of PTT, TDT, and CDT. The tumor volume of the treatment group was merely 13.3% of the control group, and the system also exhibited excellent biocompatibility. Collectively, it offers a feasible and promising intelligent platform for precise local treatment of pancreatic cancer.

<h4>Background/objectives</h4>Emerging evidence shows long non-coding RNAs (lncRNAs) as critical regulators of osteoarthritis (OA) progression, often acting in complex networks with microRNAs (miRNAs). In our study, we investigated the potential regulatory function of the lncRNA MEG3/miR-21-5p axis in the OA phenotype of chondrocytes.<h4>Methods</h4>Differential gene expression analysis in damaged vs. intact cartilage was performed, re-analyzing existing public RNA-seq data. MiRTarBase, LncRNADisease, and Open Targets databases were utilized to identify miR-21-5p target genes and OA-associated lncRNAs and genes. Functional enrichment analysis and protein-protein interaction (PPI) network construction were performed using the DAVID and STRING databases, respectively. MEG3, miR-21-5p, SOX5, COL2A1 and ACAN mRNA expressions were assessed by qRT-PCR. The role of the MEG3/miR-21-5p axis in OA chondrocytes was examined using transfection experiments.<h4>Results</h4>Eighty-one lncRNAs displayed significant differences in expression between damaged and intact cartilage, including MEG3. Bioinformatic analysis indicated that MEG3 interacts with miR-21-5p, while SOX5 was identified to be a putative target of miR-21-5p. MEG3 and SOX5 expression levels were significantly downregulated in OA chondrocytes, whereas miR-21-5p expression was upregulated. Silencing of MEG3 resulted in increased miR-21-5p levels in chondrocytes. Conversely, inhibition of miR-21-5p led to increased SOX5 expression and anabolic markers COL2A1 and ACAN. Notably, MEG3 silencing significantly reduced SOX5 expression, an effect that was reversed upon miR-21-5p inhibition.<h4>Conclusions</h4>Our findings highlight a potential regulatory role of the dysregulated MEG3/miR-21-5p axis in modulating the anabolic phenotype of chondrocytes through regulation of SOX5 expression. This novel lncRNA/miRNA/mRNA regulatory network may represent a candidate therapeutic axis for knee osteoarthritis.

<h4>Background</h4>Narcolepsy is a neurological sleep disorder associated with immune response. However, the autoimmune basis for narcolepsy remains unclear. This study aimed to evaluate the causal relationship between immune cells and narcolepsy type 1 (NT1).<h4>Methods</h4>We used a two-sample Mendelian randomization (MR) method to investigate the associations between 731 immune cell traits and NT1 based on a genome-wide association study (GWAS) database from the FinnGen consortium. The inverse-variance weighted (IVW) method was used as the primary method, followed by sensitivity analyses, including the MR-Egger intercept test, Cochran's Q test, and MR pleiotropy residual sum and outlier (MR-PRESSO). Additional mediation analysis was conducted to investigate the mediating effect of 91 cytokines on immune cells to facilitate the immune processes in NT1.<h4>Results</h4>Immune cell traits showed significant causal associations with NT1. Risk-associated traits mainly involved human leukocyte antigen-DR (HLA-DR)-related monocyte phenotypes, T-cell-related traits, natural killer (NK) cell-related traits, and natural killer T (NKT) cell-related traits, whereas protective traits mainly involved CD4+ T-cell-related, plasmacytoid dendritic cell-related, monocyte-related, and B-cell-related phenotypes. Overall, ten immune cell traits were associated with an increased risk of narcolepsy, whereas five were associated with a reduced risk. Mediation analysis further indicated that interleukin-6 (IL-6) mediates the immune-inflammatory pathway linking monocyte-related traits to NT1. These findings remained consistent in all sensitivity analyses.<h4>Conclusions</h4>Our study identified immunophenotypes that are related to the development of NT1, providing insight into the autoimmune pathogenesis of NT1 and subsequent immunotherapy.

Unexplained infertility (UI) accounts for an estimated 15-30% of infertility but its biological basis remains poorly defined. Plasma haemostatic abnormalities have been implicated in recurrent pregnancy loss; however, whether the circulating coagulation-fibrinolysis axis differs between women with UI and those with infertility attributable to other causes has not been characterised. The aim of this study was to compare 20 coagulation- and fibrinolysis-related proteins between women with UI and women with male-factor infertility (MFI) as control. In this exploratory case-control study, plasma from 11 women with UI and 14 women with MFI were analysed using SOMAscan proteomics. Group means were compared using Welch's two-sample t-test, Cohen's d with 95% confidence intervals, and a two-sided permutation test on the mean difference (20,000 random label permutations). False discovery rate (FDR) was used for multiple comparisons. Pairwise Pearson and Spearman correlations were performed. Three proteins differed between groups (<i>p</i> < 0.05), all lower in UI than in MFI: vitamin K-dependent protein S (Cohen's d = -1.11, 95% CI -1.95 to -0.26; Welch <i>p</i> = 0.016; permutation <i>p</i> = 0.015), antithrombin-III (d = -1.00, 95% CI -1.84 to -0.16; <i>p</i> = 0.024; permutation <i>p</i> = 0.020) and α2-antiplasmin (d = -0.82, 95% CI -1.64 to 0.00; <i>p</i> = 0.051; permutation <i>p</i> = 0.054). No protein survived FDR correction. Within UI, the three proteins co-varied positively, with significant pairwise correlations between protein S and antithrombin-III (r = 0.67, <i>p</i> = 0.024) and between antithrombin-III and α2-antiplasmin (r = 0.71, <i>p</i> = 0.015). Within MFI, antithrombin-III and α2-antiplasmin remained tightly correlated (r = 0.70, <i>p</i> = 0.006) but protein S decoupled from both (r = 0.20 and 0.10, both non-significant). In this hypothesis-generating study, women with UI showed reduction in three plasma regulators of anticoagulant and antifibrinolytic capacity, protein S, antithrombin-III and α2-antiplasmin relative to women with MFI, raising the possibility that UI may be in part a subtle immunothrombotic/endothelial implantation disorder.

<title>Abstract</title> <p>Background Patients with autoimmune diseases (ADs) have an elevated risk of venous thromboembolism (VTE), but the shared genetic basis remains systematically unexplored. This study aims to elucidate the shared genetic architecture and biological pathways linking ADs to VTE. Methods We conducted a cross-trait genetic analysis using GWAS summary statistics for VTE and 16 ADs. LDSC and HDL estimated genetic correlations, while PLACO and MAGMA identified shared pleiotropic loci and genes. Mendelian randomization (MR) evaluated causal links, complemented by colocalization and SMR to identify regulatory mediators. Drug-gene interaction analysis prioritized potential therapeutic targets. Results Significant positive genetic correlations were found between VTE and five ADs: rheumatoid arthritis, systemic lupus erythematosus (SLE), Sjögren's syndrome, psoriasis, and Hashimoto's thyroiditis. We identified 21 immunothrombotic shared loci and 274 pleiotropic genes, primarily enriched in fibrin clot formation and inflammatory signaling. Integrative analysis highlighted IL6R and PLCL1 as key tissue-specific mediators. MR analysis supported a causal effect of SLE on VTE risk (OR = 1.018, P = 0.0003). Drug prioritization identified TNF inhibitors and warfarin as candidates for drug repurposing in comorbid conditions. Conclusions This study provides a robust genetic foundation for the autoimmune-thrombotic axis. By characterizing "Immunothrombosis Gene Modules," our findings offer novel insights for risk stratification and targeted preventive strategies for high-risk patients with autoimmune diseases.</p>

Latent Epstein-Barr virus (EBV) infection is associated with multiple lymphoid and epithelial cancers in humans. Targeting EBV through lytic induction therapy represents a potential strategy for treating virus-associated malignancies, such as EBV-associated gastric cancer (EBVaGC). Despite its classification as a distinct gastric cancer subtype, precision therapeutic strategies for EBVaGC remain vastly underexplored. In a recent clinical study, the combination of an orally administered histone deacetylase (HDAC) inhibitor, nanatinostat (NSTAT) with valganciclovir (VGCV), showed promise as a lytic induction therapy for EBV-positive lymphoma. In this study, we evaluated the activity of NSTAT to induce EBV lytic reactivation and the therapeutic efficacy of NSTAT-based lytic induction therapy in two representative EBVaGC cell lines in vitro and in vivo. NSTAT efficiently induced the expression of EBV immediate-early, early and late genes in EBVaGC cells. In addition, NSTAT treatment also promoted global histone acetylation and suppressed c-MYC and BCL2 expression, leading to cell cycle arrest and cell death in the EBVaGC tumor cells. Importantly, this study demonstrated the potent antitumor efficacy and safety of combined NSTAT and ganciclovir (GCV) treatment in both in vitro and in vivo preclinical EBVaGC models.

<h4>Background</h4>Glucocorticoids(GCs) are widely used to treat erythropoietin-resistant anemias, yet the precise mechanisms underlying their erythropoiesis-promoting effects remain incompletely understood.<h4>Methods</h4>This study used single-cell RNA sequencing, ATAC-seq, ChIP-seq, RNA-seq,quantitative PCR (qPCR), enzyme-linked immunosorbent assay (ELISA) and flow cytometry in vivo models (AIHA patients, CD163^-/- mice, Gypa-eGFP-cremice, Epor-tdtomato-cre mice, and Epor-eGFP-cre rats) and in vitro human erythroblastic island(EBI) formation and EBI enrichment and cytospins, Giemsa and Prussian blue staining, quantification and co-culture systems to delineate CD163⁺ macrophages coordinating erythroblastic island formation and iron metabolism.<h4>Results</h4>GC promote erythropoiesis by regulating CD163-mediated EBI formation and modulating iron metabolism within EBI macrophages, a phenomenon conserved across humans, rats, and mice. We demonstrated that CD163⁺macrophages-but not their CD163^- counterparts-exhibit heightened iron metabolism in the bone marrow, and that GC-induced erythropoiesis is markedly attenuated in CD163-deficient mice due to disrupted EBI architecture and impaired iron handling. Importantly, GC therapy restores iron metabolism and mitigates inflammatory responses in BM CD163⁺macrophages, likely contributing to improved erythropoiesis in patients with autoimmune hemolytic anemia.<h4>Conclusions</h4>CD163⁺macrophages support GC-induced erythropoiesis by coordinating erythroblastic island formation and iron metabolism. These findings uncover a previously unrecognized GC-CD163-EBI axis that governs erythropoiesis and highlight the potential of targeting EBI macrophage function as a novel therapeutic strategy for anemia.

<b>Background/Objectives</b>: To evaluate the diagnostic performance of ultrasound-guided attenuation parameter (UGAP) for the assessment of hepatic steatosis in a population at risk for metabolic dysfunction-associated steatotic liver disease (MASLD), using MRI proton density fat fraction (PDFF) as the reference standard, and to also derive optimal population-specific diagnostic thresholds. <b>Methods</b>: In this single-center prospective study, 64 adults at risk for MASLD underwent UGAP measurement and MRI-PDFF. UGAP was performed according to standardized manufacturer-recommended protocols and standardized on the right hepatic lobe. Hepatic steatosis was staged using established MRI-PDFF thresholds. Diagnostic performance was evaluated using receiver operating characteristic (ROC) analysis. Cohort-UGAP cut-offs were derived using the Youden index. Associations between UGAP and clinical parameters were assessed using correlation and regression analyses. <b>Results</b>: UGAP correlated strongly with MRI-PDFF (ρ = 0.82, <i>p</i> < 0.001). The areas under the ROC curve (AUCs) for detecting mild, moderate, and severe steatosis were 0.86, 0.96, and 0.96, respectively. Right-lobe acquisitions outperformed left-lobe measurements, while four-region averaging yielded the highest diagnostic performance. UGAP values were associated with BMI, waist circumference, and liver enzymes. <b>Conclusions</b>: UGAP provides an accurate noninvasive assessment of hepatic steatosis, demonstrating high overall diagnostic agreement with MRI-PDFF. Right-lobe acquisition and multi-regional averaging further improve its performance. While cohort-specific threshold optimization may enhance clinical applicability, larger studies are needed to fully confirm its accuracy in advanced stages.

Plant-derived additives are increasingly explored as functional aquafeed ingredients for supporting fish performance and physiological condition. This study evaluated the effects of dietary <i>Wedelia chinensis</i> extract (WCE) on growth performance, feed utilization, antioxidant status, innate immune-associated biomarkers, and immune- and antioxidant-related gene expression in Nile tilapia (<i>Oreochromis niloticus</i>). Juvenile fish with an initial body weight of 14.31 ± 0.03 g were fed diets supplemented with WCE at 0, 5, 10, 15, or 20 g kg^-1 diet for eight weeks. Growth indices, somatic indices, serum and hepatic antioxidant biomarkers, mucosal and systemic innate immune-associated parameters, and liver and intestine gene expression were assessed. Dietary WCE improved final weight, weight gain rate, specific growth rate, and feed conversion ratio compared with the control, with WCE10 showing the most consistent growth-related response. Somatic indices were not significantly affected. WCE supplementation increased antioxidant enzyme activities and reduced malondialdehyde levels in serum and liver, particularly at moderate inclusion levels. It also increased lysozyme and peroxidase activities, alternative complement activity, phagocytic index, and respiratory burst activity, and modulated selected immune- and antioxidant-related genes in the liver and intestine. Taken together, dietary WCE, particularly at 10 g kg^-1 diet, improved growth and feed utilization while supporting antioxidant status and basal innate immune-associated biomarkers during the eight-week feeding trial, without detectable changes in somatic indices. These findings provide a basis for further evaluating WCE as a plant-derived functional additive for Nile tilapia aquafeeds under pathogen-challenge and practical farming conditions.

Protein-based biosensors require controlled and site-selective functionalization strategies to enable stable and oriented immobilization without compromising protein structure and signal transduction efficiency. We evaluated a chemoselective linchpin-directed modification (LDM) approach targeting Lys-His pairs as a tool for site-specific labeling of the model fluorescent biosensor green fluorescent protein (GFP). LDM molecules with variable spacer lengths were prepared, and a structure-guided computational workflow was implemented to map Lys-His distances on the protein and identify candidate modification sites. Experimental validation by UV-Vis spectroscopy and mass spectrometry demonstrated efficient conjugation and a final degree of labeling close to unity, consistent with single-site modification, with all LDM molecules selectively targeting the same histidine residue (His181), independently of spacer length. Structural analysis revealed that this residue is located within an accessible internal cavity that favors productive interactions with the reactive group. Importantly, the modification preserves GFP fluorescence and pH response, confirming retention of sensing functionality. These results demonstrate that LDM enables selective modification not only of surface residues, but also of structurally guided, non-surface residues. This approach provides the proof of concept of a new, promising strategy for the controlled functionalization and immobilization of protein-based biosensors.

Neurodegenerative diseases (NDs), such as Alzheimer's disease (AD), Parkinson's disease (PD), Amyotrophic lateral sclerosis (ALS), and Huntington's disease (HD), involve the gradual loss of structure or function of neurons in the nervous system and are an increasing threat to the aging population worldwide. Although these disorders have different clinical features which affect cognition, movement and other vital body functions, they share key underlying molecular and cellular processes. This starts with protein misfolding and aggregation, mitochondrial dysfunction, oxidative stress, dysregulated protein homeostasis, neuroinflammation, and disrupted cell death pathways. Recent findings have added disease-specific processes, like amyloid-β and tau aggregates in AD, α-synuclein aggregation and mitophagy failure in PD's, TDP-43-related impaired RNA metabolism in ALS, and mutant huntingtin causing transcription aberrations in HD. Protein interactome network analysis showed mechanistic crosstalk between pathogenic proteins of AD and PD. New evidence highlights how lysosomal dysfunction, endoplasmic reticulum stress, and microglial activation, act as a common axis in neurodegeneration. Advancements in genomics and epigenomics have found shared genetic risk loci and regulatory processes that affect how diseases develop and progress. Simultaneously, new biomarkers like circulating microRNAs, exosome-related pathological proteins, neurofilament light chain, inflammatory cytokines, and microglial activation markers are powering early diagnosis tools and disease variations. New imaging techniques also allow for the identification of protein aggregations before symptoms appear. Overall, these findings are accelerating targeted treatments and personalized medicine aimed at disease progression. This review highlights current insights into the molecular mechanisms of NDs and discusses new biomarkers and treatment targets that help future diagnostic and treatment strategies.

<h4>Objective</h4>We aimed to explore the association between thyroid indicators and pre-sarcopenic obesity, pre-sarcopenia, and visceral obesity in a community-based euthyroid cohort.<h4>Methods</h4>A total of 1280 euthyroid residents (42.3% men) aged over 40 years were recruited in 2013-2014 and re-examined in 2015-2016. Thyroid indicators were analyzed. Visceral fat area (VFA) was quantified using magnetic resonance imaging and skeletal muscle mass was calculated by bioelectrical impedance analyzer. Pre-sarcopenic obesity was defined as the coexistence of pre-sarcopenia and visceral obesity.<h4>Results</h4>During a mean follow-up of 2.1 years, 127 (14.8%), 156 (21.3%), and 120 (20.1%) participants developed new-onset pre-sarcopenic obesity, pre-sarcopenia, and visceral obesity, respectively. Compared with those with the lowest tertile of baseline serum fT3 levels, participants with the highest tertile had an elevated risk of pre-sarcopenic obesity (risk ratio [RR] 1.82, 95% confidence interval [CI] 1.20-2.76) and pre-sarcopenia (RR 1.49, 95%CI 1.04-2.15). Moreover, per 1 standard deviation increase in serum fT3 levels over 2.1 years was positively associated with incident pre-sarcopenic obesity (RR 1.40, 95%CI 1.28-1.53) and pre-sarcopenia (RR 1.29, 95%CI 1.18-1.40), respectively.<h4>Conclusion</h4>Elevated serum fT3 levels were associated with increased risks of pre-sarcopenic obesity and pre-sarcopenia in euthyroid Chinese participants.

High-resolution spatial transcriptomics requires computational methods to accurately assign transcripts to individual cells. We present SMURF (Segmentation and Manifold UnRolling Framework), a cross-platform soft-segmentation algorithm that uses deep learning to map mRNAs from capture spots to nearby nuclei. SMURF also unrolls complex tissue architectures by projecting cells onto Cartesian coordinates, enabling analyses of cell-type organization and gene expression gradients. We show that SMURF assigns mRNAs to single cells more accurately than existing approaches and robustly unrolls complex tissues to reveal zonated transcriptional programs and cell-type organization across multiple tissues and spatial transcriptomic technologies. To showcase the biological insights enabled by SMURF, we segment over 400,000 cells from the mouse ileum using Visium HD data. We identify zonated gene expression programs along the maturing intestinal villus and the transcription factors that regulate them. Importantly, we show that gene expression gradients along the proximal-distal axis of the intestine accumulate in the upper villus and that upper villus gene expression is reprogrammed by environmental signals in the lumen, suggesting that environmental inputs are major determinants of regional transcriptional identity. Together, these results establish SMURF as a powerful framework for analyzing gene expression of cells within native tissue contexts.

Mitochondria‑endoplasmic reticulum contact sites (MERCs) are dynamic, nanoscale membrane domains that serve as crucial signaling hubs for inter‑organellar communication. These specialized interfaces are maintained by a complex network composed of tethering, promoter, and disruptor proteins and coordinate a wide range of cellular processes, such as calcium and zinc ion homeostasis, lipid biosynthesis and transfer, redox signaling, mitochondrial dynamics (fission, fusion and mitophagy), autophagy, apoptosis, inflammation and cellular senescence. Accordingly, the structural and functional integrity of MERCs is vital for cellular adaptation and survival. Nevertheless, MERC plasticity is often impaired in various human pathologies. Alterations in MERC composition, abundance, or function are regarded as pathogenic mechanisms in neurodegenerative diseases, metabolic disorders, cardiovascular conditions, cancer and orthopedic diseases. Common manifestations of MERC dysfunction include disrupted ion signaling, bioenergetic failure, excessive oxidative stress, and impaired organelle quality control. Therefore, targeted modulation of MERCs represents a promising therapeutic avenue. However, translating this potential into clinical practice faces considerable challenges. This is because MERC function is dynamic, context‑dependent and dualistic; both excessive and deficient coupling can drive pathology. Future progress hinges on deciphering the precise regulatory codes that govern MERC assembly, developing tools for real‑time, high‑resolution in vivo analysis, and designing innovative, cell‑type‑specific interventions that normalize rather than simply inhibit or enhance MERC function. A multidisciplinary approach integrating spatial proteomics, super‑resolution imaging, and advanced disease modeling is warranted for unlocking the full diagnostic and therapeutic potential of these organelle contact sites.

Loss of SMARCB1/INI1 expression is a common feature of various types of tumors including malignant rhabdoid tumor, an aggressive cancer of children. Because SMARCB1/INI1 deficiency leads to genome-wide transcriptional dysregulation, identifying a specific therapeutic target that acts on a wide range of SMARCB1/INI1-deficient tumors has been challenging. This study aimed to establish if the personalized selection of effective drugs against SMARCB1/INI-deficient tumors is possible by in vitro drug sensitivity profiling. The in vitro drug sensitivity profiles of tumors from SMARCB1/INI1-deficient tumor cell line-derived mouse xenograft (CDX) models were evaluated by a short-term collagen gel-embedded three-dimensional culture-drug sensitivity test (3D-DST). With the 3D-DST, molecular targeting drugs, including inhibitors of aurora B kinase, mammalian target of rapamycin, mitogen-activated protein kinase, WNT/β-catenin, or mouse double minute 2 homolog (MDM2), were selected as therapeutic drug candidates in SMARCB1/INI1-deficient tumors. A CDX tumor, which was resistant to MDM2 inhibitors in the 3D-DST, expressed lower TP53 compared to an MDM2 inhibitor-sensitive tumor. In cultured cell lines, exposure to MI-773, a MDM2 inhibitor, disturbed the expression of a significant number of genes and induced p21 protein expression in MDM2 inhibitor-sensitive cells. However, such changes were not observed in resistant cells. These results suggest that 3D-DST can predict biologically relevant drug sensitivity profiles in SMARCB1/INI1-deficient tumors.

Inflammation is a finely tuned host defense mechanism whose perpetual activation is a driver, promoter, and supporter of carcinogenesis. Key mediators of chronic inflammatory processes, <i>viz.</i> NF-kB, JAK-STAT, inflammasomes, reactive oxygen species (ROS), and cytokine network, if left unanswered, foster the tumour-supportive environment. Within the tumour microenvironment (TME), inflammatory cells, in combination with stromal and cancerous cells, modulate these pathways and regulate critical tumour hallmarks. Therefore, efforts have been made to understand and tackle the interface between the inflammation-cancer axis, but therapeutic outcomes remain limited. In this context, integrating systems-level biological insights with precision-driven medicinal chemistry may pave the way towards next-generation anti-inflammatory chemotherapeutics. The current review underlines the critical involvement of inflammation in cancer development by providing a comprehensive overview of key molecular pathways. A special emphasis was placed on understanding the medicinal chemistry campaign over the last 5 years for the development of inflammation-targeting small-molecule therapeutics in cancer.

Age-related vestibular dysfunction (ARVD) is a prevalent and debilitating condition among the elderly, yet its etiology and underlying molecular mechanisms remain poorly understood. We focused on mechanosensitive hair cells (HCs), which are widely recognized as susceptible to aging. Using single-cell RNA-seq transcriptomic analysis of young and old mice, we show that old vestibular HCs exhibit conserved transcriptomic hallmarks of cellular aging, including cellular senescence, mitochondrial dysfunction, and impaired proteostasis, along with prominent cell type-specific changes linked to hair bundle architecture and the mechanotransduction machinery. Consistent with these transcriptomic findings, imaging and electrophysiological recordings from old vestibular sensory epithelia reveal hair bundle degeneration and reduced mechanotransduction activity. Importantly, this structural and functional deterioration precedes HC loss, underscoring impaired hair bundle function as a key driver of ARVD. Furthermore, our comparative analysis identifies both shared and distinct aging signatures in vestibular and cochlear HCs, providing broader insight into the mechanisms that may underlie their different rates of age-related degeneration.

Multidrug resistance (MDR) remains the core clinical barrier limiting the achievement of durable and effective treatment for various tumors. With the deepening of understanding, MDR has been redefined from the traditional, static "drug efflux" model to a systemically adaptive survival strategy driven by highly heterogeneous tumor populations under therapeutic pressure, characterized by dynamic evolution. This complex phenotype arises from the deep intertwining and synergistic action of multi-layered survival networks. Herein, this review systematically delineates the evolution and multidimensional remodeling of the underlying mechanisms of MDR. We comprehensively outline the cell-intrinsic, autonomous resistance mechanisms of tumor cells, including intracellular drug redistribution and evasion of non-apoptotic cell death pathways. Concurrently, we summarize the tumor microenvironment (TME)-mediated, non-autonomous resistance mechanisms, such as physical barriers formed by stromal cells, intercellular communication networks, and the establishment of a profoundly immunosuppressive microenvironment. Building on this foundation, the review critically assesses the current drug resistance dilemmas faced by various therapeutic modalities and refractory cancer types. It focuses on discussing novel, precision reversal strategies targeting MDR, including nanotechnology-based delivery systems, single-cell and spatial omics analysis, and artificial intelligence (AI) large model-driven predictive and interventional systems. Furthermore, this review explores the underlying reasons for the repeated clinical failures of traditional single-target interventions. It outlines a direction for future translational research: a paradigm shift from "singular target killing" to "multidimensional ecological remodeling," advancing towards a closed-loop, personalized precision therapy framework based on dynamic monitoring and multi-target synergistic intervention.

Spheroids, organoids, and further three-dimensional cellular models represent an intermediate stage of supra-cellular complexity between monolayered cell cultures and animal models. In the present review, we identified conditions in which spheroids and organoids could replace animal models in biomedical research, using long noncoding RNA (lncRNA) research in cancer as a study object. In tumor spheroids and patient-derived organoids, chemosensitivity can be enhanced through therapeutic overexpression or silencing of specific lncRNAs (depending on the lncRNA and context), which then decreases spheroid/organoid size or formation efficiency. These outcomes consistently mimic observations in xenograft mouse models, in which the same lncRNA intervention decreases tumor volume. Therefore, as a proxy of tumor growth assessment, animal models could be largely replaced with spheroids or organoids, taking advantage of their inexpensiveness and patient-specific features, respectively. The use of animal models could be restricted to bystander or pleiotropic effects, such as the assessment of parameters including metastasis and survival rates.

<h4>Background</h4>The rising global health crisis of childhood overweight and obesity is potentially influenced by caesarean delivery (CD), but it remains a subject of ongoing debate. The gut microbiome, which is affected by delivery mode and can impact body weight, might play a role in this issue. However, the complex relationship between them remains poorly understood.<h4>Methods</h4>We analysed data from a randomised, double-blind, placebo-controlled trial VDAART cohort, including BMI percentiles from 683 children aged 2-8 years and 1672 stool samples collected between 3 months and 5 years (all data in this study were collected between May 2010 and February 2018). To evaluate how CD relates to BMI trajectories, we conducted permutation testing and discussed the effect of confounding factors. We then used PERMANOVA, random forest classification, and Generalised Microbe Phenotype Triangulation (GMPT) to explore the role of the gut microbiota in mediating this relationship.<h4>Findings</h4>Compared with vaginal delivery, intrapartum CD (iCD) rather than antepartum CD (aCD) was associated with a higher BMI percentile trajectory (Δ = 31.8%; 95% CI, 16.25%-47.55%; P = 0.001, Permutation test), and this was observed only among female children. Moreover, delivery mode was significantly associated with early-life gut microbiota, with effects also limited to females (F = 2.15 and 2.47 at months 3-6 and at age 1; P = 0.035 and 0.007, PERMANOVA). Random Forest models using early microbiota data can predict later overweight/obesity, performing best among iCD-born females (AUROC = 0.88; 95% CI, 0.83-0.94 for age 2), indicating an optimal intervention window before age one. Finally, GMPT identified 24 early-life taxa potentially mediating iCD-related overweight/obesity risk (11 preventive; 13 permissive), including Bacteroides ovatus, Bifidobacterium bifidum, Clostridium leptum, Eggerthella lenta, etc. INTERPRETATION: Our results indicate that CD types and children's sex are key factors in this interaction, offering a possible explanation for the ongoing debate about whether CD is linked to childhood overweight/obesity, and providing valuable insights for future intervention strategies.<h4>Funding</h4>This work was supported by the National Institutes of Health.

Colorectal cancer (CRC) is a common global malignancy, and its advanced stage is closely linked to a gut microbiota-metabolism-immunity vicious cycle requiring early intervention. In this study, anti-CRC effects of PAMK in CT26 tumor-bearing mice were explored. Results showed that PAMK significantly inhibited tumor growth and improved the quality of life of tumor-bearing mice by enhancing antitumor immunity, including increased NK cell infiltration and NKG2D expression, elevated CD4⁺:CD8⁺ ratios, and higher serum IFN-γ levels. However, therapeutic effects of PAMK were not observed in antibiotic‑induced microbiota depletion (AIMD) mice. Notably, following fecal microbiota transplantation (FMT), therapeutic effects of PAMK were largely restored. PAMK alleviated tumor-induced gut microbiota dysbiosis characterized by enriched g_Alistipes, and remodeled fatty acid and steroid metabolism, which was closely associated with enhanced antitumor immunity and a potential microbiota-metabolism-immunity axis. Meanwhile, PAMK modulated multiple metabolic, circadian and immune pathways in the spleen as verified by transcriptomics and qPCR. Integrative multi-omics analysis indicated that the gut microbiota-metabolite-spleen gene axis may act synergistically to mediate anti-CRC effects of PAMK in tumor-bearing mice. This study highlights the potential of PAMK in CRC tumor immune adjuvants, providing experimental and theoretical support for its clinical translation and novel tumor immunotherapies.

Cardiovascular disease remains the leading cause of global mortality. Understanding its complexity requires dissecting the heart's cellular landscape. Here we present HeartMap, a comprehensive single-nucleus RNA sequencing atlas of the adult human heart. This resource integrates data from nine studies, encompassing over 2.4 million nuclei, 209 individuals, eight anatomical regions and seven disease and healthy states. After rigorous data harmonization and method comparison of batch correction methods, we characterized transcriptional diversity across 14 cell types. To demonstrate the utility of HeartMap, we identified robust disease-associated gene signatures in dilated cardiomyopathy by comparing multiple studies. Notably, we identified distinct activated fibroblast populations, enriched for COL22A1 or TNC, that display variable prevalence across cardiomyopathies. HeartMap provides a valuable tool for exploring cardiac disease at the single-cell level, facilitating both fundamental research and potential therapeutic development.

Huntington's disease (HD) is a hereditary movement disorder caused by a CAG repeat expansion in the huntingtin gene. HD is characterized by deposition of mutant huntingtin (mHTT) aggregates, and by severe neurodegeneration of the basal ganglia and neocortex. No cure is currently available, and new treatment options are urgently needed. Here, we show that the oligomer modifying molecule anle138b (INN: emrusolmin) improves multiple disease phenotypes in cell culture and in two mouse models of HD. Application of anle138b reduced mHTT aggregate formation and ameliorated neurotoxicity in primary neurons. Oral administration of anle138b delayed deposition of mHTT inclusions, reduced brain atrophy, mitigated neuroinflammation and transcriptional alterations, improved motor function and extended life span in HD mice. Downregulation of striatal markers and synapse loss in striatal spiny projection neurons were also partially rescued. No adverse effects of anle138b were observed in wildtype animals. Moreover, anle138b markedly decreased mHTT aggregation in human neural precursor cells differentiated from HD patient-derived induced pluripotent stem cells (iPSCs). Altogether these results illustrate the potential of anle138b as a disease-modifying treatment for HD.

African swine fever virus (ASFV) encodes numerous multigene family (MGF) proteins whose individual contributions to virulence and immune modulation remain poorly understood. Here, we characterize the role of MGF110-2L in shaping innate immunity and disease outcome <i>in vivo</i>. Deletion of MGF110-2L from the virulent Arm/07/CBM/c2 isolate markedly attenuated clinical disease and conferred complete protection against homologous challenge. In macrophages, the MGF110-2L-deficient recombinant induced enhanced type I interferon (IFN) responses, consistent with a role for MGF110-2L in antagonizing antiviral signaling. However, transcriptomic profiling of peripheral blood mononuclear cells revealed that the absence of MGF110-2L prevents the excessive IFN and inflammatory hyperactivation triggered by the parental virus <i>in vivo</i>, establishing a controlled antiviral program associated with reduced pathology. These findings identify MGF110-2L as a functional virulence factor that drives dysregulated innate immune responses during ASFV infection. Our work advances the molecular understanding of ASFV-host interactions and highlights the contribution of MGF110-2L protein to immune evasion and pathogenesis.IMPORTANCEFinding safe and effective vaccines is essential to impair the spread of African swine fever virus (ASFV), responsible for the largest animal pandemic. Identifying ASFV virulence factors and understanding the mechanisms of pathogenesis and protection are crucial. By using RNAseq from <i>in vivo</i> peripheral blood mononuclear cells, together with immunological studies, we showed that MGF110-2L regulates type I IFN production and cytokine storm in pigs. Deletion of the MGF110-2L gene results in attenuation and induces protection against the parental virulent strain. These results support the induction of an alert immune state together with dampened systemic inflammation but preserved cytotoxic readiness in vaccinated animals. Altogether, our data shed light on the mechanisms underlying protection against ASFV and contribute to the development of new protective tools.

SUMOylation is a post-translational modification in which a Small Ubiquitin-like Modifier (SUMO) protein is reversibly attached to a lysine residue on a target protein in an ATP-dependent process. This modification can affect the function of target proteins by enhancing their stability or changing cellular translocation, thereby making SUMOylation a critical regulator in the pathogenesis of multiple diseases. The functional consequences of SUMOylation, however, are highly context dependent. In Alzheimer's disease, SUMOylation stabilizes proteins that drive disease progression and enhances neurotoxicity, thereby exacerbating these conditions. Similarly, in Progressive Supranuclear Palsy, SUMO-1 conjugation stabilizes truncated tau and blocks its ubiquitination, whereas SUMO-2/3 conjugation promotes Tau clearance and recovery from neuroinflammation, illustrating how distinct SUMO paralogues can exert opposing effects within the same disease. Conversely, increased SUMOylation can be neuroprotective in cerebral ischemia and Parkinson's disease by promoting autophagic clearance of pathogenic proteins. Beyond alterations in protein stability, aberrant SUMOylation can also lead to mis-localization of target proteins, which has been identified as a pathogenic mechanism in disorders such as Huntington's disease and Amyotrophic Lateral Sclerosis that results in impaired clearance and pathogenic buildup, which results in neuronal death. From a therapeutic standpoint, the SUMO inhibitor TAK-981 has shown promise in both Multiple Sclerosis and in pre-clinical glioblastoma models, underscoring the translational potential of targeting of this pathway. This review examines the multifaceted role of SUMOylation across diverse neurological conditions, evaluates the therapeutic potential of SUMO inhibitors and activators, and highlights the opportunities and challenges of modulating this pathway in currently incurable neurological disorders.

Osteosarcoma (OS) suffers from stagnant survival rates due to high metastatic potential and genomic complexity. Here, we present a high-resolution single-cell transcriptomic atlas of the OS ecosystem using single-cell fixed RNA profiling (FLEX) of 40,401 cells across four clinical stages: treatment-naïve, post-chemotherapy, recurrent, and lung metastatic. We identified a metastasis-enriched malignant subpopulation, OB_3, characterized by aggressive transcriptional signatures and governed by <i>ADAMTS12,</i> which we validated as a prognostic marker for reduced overall survival. Functional <i>in vitro</i> experiments further demonstrated that <i>ADAMTS12</i> silencing significantly reduced the migratory capacity of HOS and 143B OS cells and suppressed AKT and ERK phosphorylation, thereby supporting its role as an active driver of metastatic progression. Within the microenvironment, metastatic progression was defined by the emergence of RNASE1+ M2-like tumor-associated macrophages, an expansion of highly immunosuppressive S100A4+ regulatory T cells, and the specialization of PLVAP<b>+</b> stalk endothelial cells driving tumor angiogenesis. Integrative interactome analysis revealed that these populations function as central coordinating hubs, utilizing MIF, SPP1, and Galectin signaling to orchestrate the metastatic niche. Our findings, validated across multiple external cohorts and tissue microarrays, delineate a coordinated multi-cellular network involving ADAMTS12+, RNASE1+, S100A4+, and PLVAP+ cells. This study clarifies how malignant and microenvironmental components co-evolve to facilitate systemic dissemination, providing a translational framework for precision risk stratification and the development of next-generation therapeutic strategies to counteract metastatic OS.

Xylosyltransferase-II (XT-II), encoded by <i>XYLT2</i>, catalyzes the rate-limiting initial step of proteoglycan biosynthesis. Pathogenic mutations in <i>XYLT2</i> cause spondylo-ocular syndrome (SOS), a rare disorder characterized by severe primary osteoporosis, skeletal dysplasia, and additional systemic manifestations. Although dysregulated macrophage polarization has been linked to bone remodeling disorders such as osteoporosis, which are associated with an imbalance toward pro-inflammatory and osteoclastogenic signaling, the role of XT-II in macrophage biology remains unexplored. In the present study, siRNA-mediated knockdown of <i>XYLT2</i> was performed in human primary macrophages across three polarization states (M0, M1, M2) to investigate the impact of XT-II deficiency on macrophage polarization and inflammatory signaling. At the phenotypic level, <i>XYLT2</i> deficiency destabilized M2 macrophage identity, as shown by significant reductions in M2-associated marker expression, including interleukin 1β, interleukin 6, CD163, and CD206, and impaired phagocytic capacity. A paradoxical reduction of pro-inflammatory markers alongside the induction of M2-associated features was observed in M1 macrophages, indicating a broader disruption of polarization boundary maintenance. The <i>XYLT2</i> deficiency also promoted pro-inflammatory activation in unpolarized M0 macrophages. Cytokine profiling revealed a predominantly pro-inflammatory secretory shift, with exceptional induction of CXCL10 across all polarization states. At the signaling level, <i>XYLT2</i> knockdown resulted in a reciprocal shift, characterized by suppression of NF-κB pathway components and nuclear p65 translocation alongside the constitutive activation of STAT1 and STAT3. Transcriptome-wide profiling by bulk mRNA sequencing confirmed a conserved interferon-associated gene expression program across all polarization states, with significant enrichment of innate immune sensing, JAK-STAT, and cytokine signaling pathways among upregulated genes. In summary, these findings demonstrate that XT-II fulfills an important role in maintaining macrophage polarization and that its deficiency induces a reprogramming of pro-inflammatory signaling, extracellular matrix remodeling, and cellular interactions in bone-relevant cell types. This is associated with transcriptional changes linked to osteoclast differentiation and altered bone-associated immune signaling, suggesting that <i>XYLT2</i> deficiency may contribute to impaired bone homeostasis and providing new insights into the pathomechanisms of SOS.

<h4>Background</h4>Lower educational attainment is associated with obesity and type 2 diabetes (T2D), but prospective evidence, external validation, mediator patterns, and genetic triangulation have rarely been integrated.<h4>Methods</h4>UK Biobank analyses included 501,932 participants at baseline and 474,659 participants without baseline diabetes prospectively. Logistic and Cox models estimated associations of higher educational attainment with prevalent and incident diabetes. T2D pathway analyses evaluated adiposity, health behaviors, and cardiometabolic biomarkers. NHANES 2011-2018 provided weighted cross-sectional validation. Public GWAS summary statistics were used for genetic correlation, Mendelian randomization (MR), tissue enrichment, candidate-gene analyses, shared-locus analyses, and S-PrediXcan transcriptome-wide association analyses (TWAS).<h4>Results</h4>In fully adjusted UK Biobank models, higher educational attainment was associated with lower odds of prevalent T2D (OR 0.76, 95% CI 0.72-0.79) and lower risk of incident T2D (HR 0.70, 95% CI 0.68-0.72). Detailed qualification categories showed lower prevalent T2D odds for college/university degree versus no qualifications (OR 0.69, 95% CI 0.65-0.72). In NHANES, college graduation or above was associated with lower prevalent T2D odds (OR 0.69, 95% CI 0.61-0.78; n=20,502). Adiposity, smoking, alcohol use, lipids, blood pressure, and C-reactive protein attenuated the education-T2D association. Genetically predicted educational attainment was inversely associated with BMI and T2D, and BMI/T2D brain TWAS identified 19 shared multi-SNP gene signals, including NPC1, HSD17B12, MAP2K5, DHX36, BHMT, and LEPROT.<h4>Conclusions</h4>Higher educational attainment was consistently associated with lower T2D risk across UK Biobank, NHANES, and genetic analyses. The results highlight modifiable metabolic and behavioral pathways relevant to T2D prevention.

Recessive dystrophic epidermolysis bullosa (RDEB) is a severe skin fragility disorder caused by mutations in <i>COL7A1</i> gene, leading to chronic injury, inflammation, and debilitating sensory symptoms, including pain and itch. While structural defects in the dermo-epidermal junction are well characterized, the mechanisms underlying impaired sensory reinnervation and neuropathic manifestations remain poorly understood. Here, we investigated whether defective reinnervation in RDEB is driven by intrinsic neuronal deficits or by alterations in the cutaneous microenvironment. Using a prospective cohort of RDEB patients with small fiber neuropathy (SFN), combined with high-resolution digital PCR and multiplex cytokine profiling, we analyzed the transcriptional and secretory responses of wounded skin and primary keratinocytes. RDEB tissue exhibited a markedly blunted transcriptional response to injury, with failure to induce key inflammatory, proteolytic, and axonal guidance genes, despite a sustained pro-inflammatory secretome characterized by elevated IL-6, TNF-α, IL-1β, CCL2, and MMP9. Functionally, RDEB blister fluid induced growth cone collapse and impaired neurite outgrowth in sensory neurons. These findings reveal a dissociation between transcriptional activation and extracellular signalling, resulting in a non-permissive niche for nerve regeneration. We propose that this altered microenvironment simultaneously impairs reinnervation and promotes maladaptive nociceptive signalling, providing a mechanistic link between chronic denervation and neuropathic pain in RDEB.

Acute myeloid leukemia (AML) is one of the most common types of hematological malignancies and a leading cause of cancer deaths. It is characterized by the rapid accumulation of typically immature myeloid cells that serve to disrupt the production of mature cells, leading to a range of clinical sequelae. The role of recurrent chromosomal aberrations has long been appreciated in this disease, but a myriad of gene mutations have been increasingly acknowledged as having important roles. This review provides a comprehensive overview of the mutational landscape of AML, discussing the various genetic lesions in terms of their function, classification, etiological role, prognostic value, therapeutic impact, detection, and monitoring, with a particular focus on gene mutations.

Surface functionalization of metallic implants is widely explored to enhance their performance and functionality. In this study, multifunctional hydrogel coatings based on poly(vinylpyrrolidone) and polyethylene glycol were developed and functionalized with a taxifolin (TAX) inclusion complex and collagen to introduce bioactive features. TAX, a naturally occurring flavonoid with antioxidant and anti-inflammatory properties, was incorporated using β-cyclodextrin to improve its stability and enable controlled release. The coatings were applied to titanium-hydroxyapatite composites and titanium sheet substrates to evaluate their applicability across surfaces with varying morphologies, ranging from porous to relatively smooth. The ceramic phase was modified with magnesium ions to enhance its bioactivity and better mimic the composition of natural bone tissue. FTIR and SEM analyses confirmed hydrogel formation and effective surface coverage. Degradation and incubation studies in simulated physiological environments demonstrated the material's stability, while UV-Vis analysis indicated TAX release, highlighting the system's potential as a carrier for flavonoid-based compounds. Indirect cytotoxicity studies using MC3T3-E1 preosteoblasts indicated low cytotoxicity and a favorable biological response of collagen- and taxifolin-modified systems. The developed coatings represent a versatile platform for surface modification of titanium-based biomaterials and demonstrate potential for application across substrates with diverse surface characteristics. Further studies are required to assess their biological potential.

Triptolide (TP) is an effective anti-inflammatory and immunosuppressive agent, yet its clinical application is constrained by significant male reproductive toxicity. Curcumin, a natural antioxidant, exhibits protective effects; however, whether it protects against TP-induced damage during mouse spermatogenesis and the underlying mechanisms remain incompletely understood.<h4>Methods</h4>Proteomic analysis was performed to investigate the protective mechanism of curcumin in mouse GC-1 cells, followed by multiple validation assays including CCK-8 assay, apoptosis detection, measurement of reactive oxygen species (ROS), glutathione (GSH), malondialdehyde (MDA), and Fe²⁺ levels, quantitative polymerase chain reaction (qPCR), Western blotting (WB), hematoxylin-eosin (HE) staining, and immunofluorescence.<h4>Results</h4>Proteomic analysis revealed that curcumin primarily ameliorated TP-induced damage in mouse spermatogonia by modulating ferroptosis-related pathways. Curcumin elevated GSH levels; reduced MDA, ROS, and Fe²⁺ levels; alleviated lipid peroxidation; and regulated ferroptosis-related pathways in both TP-induced GC-1 cells and testicular tissue. These effects were associated with upregulation of the mRNA and protein expression of <i>Nrf2</i>, <i>Gclc</i>, and <i>Map1lc3a</i> and downregulation of <i>Tfrc</i> and <i>Dmt1</i>. Collectively, these findings demonstrate the protective effect of curcumin against TP-induced spermatogonial damage.<h4>Conclusions</h4>Curcumin regulated ferroptosis-related pathways by modulating the expression of <i>Nrf2</i>, <i>Gclc</i>, <i>Map1lc3a</i>, <i>Tfrc</i>, and <i>Dmt1</i>, thereby significantly ameliorating TP-induced damage in mouse spermatogonia.

Progression to fibrosis is a major complication of chronic kidney disease (CKD) in obesity, type 2 diabetes, hypertension, and metabolic syndrome, yet effective antifibrotic therapies remain limited. Here, we review how disordered renal energy metabolism-ectopic lipid accumulation, impaired fatty acid oxidation (FAO), and a compensatory shift toward glycolysis-drives tubulointerstitial fibrosis in fatty kidney disease. Lipid overload in tubular, glomerular, and vascular cells arises from increased uptake via scavenger and lipoprotein receptors, enhanced lipogenesis, and reduced lipid catabolism and clearance. Spatial lipidomic studies further reveal nephron-segment-specific lipid signatures and obesity-associated oxidized phospholipids linked to glomerular inflammation. Lipotoxicity, mitochondrial damage, and associated innate-immune signaling, ferroptosis, cellular senescence, and adipose-derived mediators (including leptin, adiponectin, and a locally active renin-angiotensin system) converge on myofibroblast activation from pericytes, fibroblasts, and other resident cells. We discuss established and emerging therapies targeting this metabolic axis-peroxisome proliferator-activated receptor-α (PPARα) modulators, sodium-glucose cotransporter-2 (SGLT2) inhibitors, glucagon-like peptide-1 (GLP-1) receptor agonists, and the mineralocorticoid receptor antagonist finerenone-and propose that restoring metabolic flexibility, by rescuing FAO while limiting maladaptive glycolysis, offers a promising disease-modifying strategy for fatty kidney disease.

<b>Background/Objective</b>: Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder characterized by motor dysfunction, cognitive impairment, and psychiatric symptoms. As the disease progresses, weight loss, cachexia, and musculoskeletal atrophy are common, reducing quality of life, decreasing their autonomy in their activities of daily living (ADLs), and increasing morbidity and mortality risk. To describe and compare energy expenditure (EE) during ADLs and resting conditions in individuals with HD and healthy controls, and to examine its associations with quality of life, cognitive status, motor function, and functional capacity. <b>Methods:</b> A cross-sectional observational study was conducted with 16 people with manifest HD and 10 healthy controls. Participants completed five ADLs: resting, dressing, combing hair, feeding, and walking under laboratory conditions. EE during ADLs was measured using a portable indirect calorimetry system. <b>Results</b>: Statistically significant between-group differences in EE were found only during feeding, with individuals with HD showing higher EE than controls (<i>p</i> = 0.021). In the exploratory correlation analysis, cognitive status was significantly associated with EE during dressing (<i>p</i> = 0.033). <b>Conclusions</b>: This exploratory study contributes to the limited evidence on EE during ADLs in adults with HD. The findings suggest that individuals with HD may expend more energy than healthy controls during specific daily activities, particularly feeding. However, these results should be interpreted with caution due to the small sample size and preliminary nature of the study. Larger, multicenter, and longitudinal studies are needed to confirm these findings and determine their clinical relevance.

Mitochondrial homeostasis is maintained by multiple quality control pathways, including mitophagy, which targets dysfunctional mitochondria for degradation. During receptor-mediated mitophagy, the outer membrane proteins BNIP3 and NIX directly recruit autophagy machinery to the mitochondrial surface, though their precise regulation is still unclear. In recent years, new BNIP3- and NIX-interacting proteins have been identified that influence mitophagic flux. PPTC7 and FBXL4 target BNIP3 and NIX for proteasomal turnover to keep levels of the receptors low, whereas TMEM11 is proposed to spatially control mitophagy by interacting with receptors at active mitophagy sites. However, it is unclear how each of these interactions is controlled and how they interplay with each other. Here, we identify a repressor of mitophagy, ARMC1, which forms a complex with TMEM11, BNIP3, and NIX. During mitophagy activation, ARMC1 dissociates from the complex, freeing the receptors to initiate mitophagy. We find that TMEM11 then acts in an antagonistic relationship with PPTC7, protecting the receptors from proteasomal degradation. Our data are consistent with a two-stage model. At steady state, a population of sentinel receptors is repressed and primed to respond to mitochondrial dysfunction. Once mitophagy is activated, TMEM11 protects BNIP3 and NIX, ensuring a sustained mitophagic response. Our findings provide a framework for understanding how two key regulatory pathways intersect to modulate receptor-mediated mitophagy.

Huntington’s disease (HD), caused by a CAG repeat expansion in the huntingtin ( HTT ) gene, is characterized by progressive neurodegeneration and accumulation of DNA damage with multiple disease-modifier genes involved in DNA repair pathways. Previous studies have implicated ataxia telangiectasia mutated (ATM) signaling in the regulation of genomic stability and DNA damage repair (DDR) pathways in HD. ATM has also been linked to the WW domain-containing oxidoreductase (WWOX), a protein involved in DNA repair and maintenance of genomic stability, through the E3 ubiquitin ligase ITCH. However, whether this signaling pathway contributes to HD pathogenesis remains unknown. Here, we investigated the role of ATM-ITCH-WWOX signaling in HD. Our results revealed no significant alterations in total ATM, phosphorylated ATM (pATM-S1981), or ITCH in HD post-mortem prefrontal cortex (PFC) compared to controls. Although treatment of human neuroblastoma SH-SY5Y cells with HD PFC lysates did not alter pATM-S1981 levels, it increased histone H2AX phosphorylation at S139 (γ-H2AX), a marker of DNA double-strand breaks. This finding suggested the presence of persistent DNA damage signaling independent of canonical ATM activation. Conversely, WWOX levels were increased in both HD PFC and HD embryonic stem cell-derived cortical neurons. Additionally, treatment of SH-SY5Y cells with recombinant human WWOX protein or WWOX overexpression increased γ-H2AX levels, supporting a role for WWOX in promoting DNA damage. To determine whether WWOX contributed to DNA damage in HD, SH-SY5Y cells were treated with HD PFC lysates that were depleted of WWOX. Immuno-depletion of WWOX reduced the ability of HD PFC lysates to increase γ-H2AX, suggesting that WWOX contributes to DNA damage in HD. Finally, overexpression of WWOX in RPE1-AAVS1-CAG115 cells did not affect somatic CAG repeat instability, despite persistent increases in γ-H2AX levels. Collectively, our findings identify WWOX as a contributor to DNA damage in HD, acting independently of the ATM pathway.

The outflow tract (OFT) of the heart carries blood away from the heart into the great arteries. During embryogenesis, the OFT divides to form the aorta and pulmonary trunk, creating the double circulation present in mammals. Defects in this area account for one-third of all congenital heart defect cases. Here, we present comprehensive transcriptomic data on the developing OFT at two distinct time points (embryonic and fetal) and its adult derivatives, the aortic valves, and use spatial transcriptomics to define the distribution of cell populations. We uncover that distinctive embryonic signatures persist in adult cells and can be used as labels to retrospectively attribute relationships between cells separated by a large timescale. Single-cell regulatory network inference identifies GATA6, a transcription factor linked to common arterial trunk and bicuspid aortic valve, as a key regulator of valve precursor cells. Its downstream network reveals candidate drivers of human cardiac defects and illuminates the molecular mechanisms of both normal and pathological valve development. Our findings define the cellular and molecular signatures of the human OFT and its distinct cell lineages, which is critical for understanding congenital heart defects and developing cardiac tissue for regenerative medicine.

The DNA damage response (DDR) is critical for pancreatic ductal adenocarcinoma (PDAC) development and therapeutic responses, including to genotoxic agents. While epigenetic modulators have been shown to contribute to the DDR, how chromatin regulation dictates responses to DNA damage in PDAC remains incompletely understood. Here, we identify Class I histone deacetylases (HDACs) as critical regulators of the DDR. HDAC1/2 direct the genomic distribution of H3K27ac, ensuring sufficient BRD4 and RNA polymerase II (Pol II) occupancy at DDR gene promoters. HDAC inhibition by entinostat shifts the balance of H3K27 acetylation preferentially toward intergenic regions, diverting BRD4 and Pol II from promoters, thereby suppressing DDR gene expression. In line with this, HDAC inhibition heightens DNA damage and sensitizes PDAC to diverse DNA-damaging and DDR-targeting agents. Since the clinical development of HDAC inhibitors has been limited by systemic toxicity, we developed bottlebrush prodrug (BPD) nanoparticles for tumor-selective entinostat delivery. Entinostat-BPD achieved tumor-specific HDAC inhibition while displaying potent efficacy and reduced systemic toxicity. These findings reveal an HDAC-dependent DDR vulnerability and offer combinational and precision targeting strategies to facilitate clinical translation and improve PDAC patient outcomes.

Fibro-adipogenic progenitors (FAPs) are muscle-resident mesenchymal stromal cells essential for homeostasis and regeneration but can produce fibrosis or intramuscular fat in pathological conditions. Insulin-like growth factor-I (IGF-I) regulates regeneration through actions on muscle fibers, satellite cells, macrophages, and extracellular matrix (ECM) remodeling but has multiple sources. To assess the role of FAP-derived IGF-I, we generated inducible FAP-specific Igf1-deficient (FID) mice. Following BaCl₂ injury, FID muscles displayed impaired regeneration, with smaller fibers, fewer Pax7⁺ and MyoD⁺ cells, increased CD68⁺ macrophages, decreased collagen, and suppressed FAP proliferation. After glycerol-induced injury, FID muscles had reduced fat. Primary FID FAPs displayed blunted proliferation, upregulated immune-regulatory genes, and downregulated ECM and growth genes, with delayed fibrogenic and adipogenic differentiation. scRNA-seq of homeostatic muscle revealed reduced protein translation and ECM indices alongside increased senescence markers in FID samples. Taken together, FAP IGF-I is critical for FAP function, with direct and indirect impact on muscle regeneration.

Cancer-associated fibroblasts (CAFs) form a dynamic ecosystem that critically influences tumor progression and therapeutic response. Although recent advances in single-cell and spatial omics have uncovered profound stromal diversity, interpreting the mechanistic relevance of this complexity remains a challenge. Here, we propose a more unifying conceptual framework to bridge high-dimensional data with experimental biology. By categorizing CAFs into conserved molecular phenotypes and distinct spatial archetypes, this model illustrates how CAF identities are intimately linked to local tissue contexts. This refined framework brings the complexity of the tumor stroma into greater focus, underscoring the necessary transition from broad stromal targeting toward precision, context-specific modulation. Ultimately, we hope this integrated effort will aid in the collaborative development of next-generation therapies that selectively target pathogenic stroma in cancer to improve patient outcomes.

To understand the mechanisms underlying the remarkable resistance of Göttingen minipigs to metabolic dysfunction-associated steatotic liver disease (MASLD), a multilevel transcriptomic analysis comparing their liver with those of humans, mice, and MASLD susceptible pig breeds was performed. This analysis revealed 692 genes uniquely differentially expressed in livers of Göttingen minipigs, linked to multiple metabolic and signaling pathways (e.g., AMPK signaling). Among these, 11 transcription factors (TFs), 3 hepatokines (DPP4, ITIH1, and ITIH3), and 6 additional secreted proteins (including PCSK9 and APOM) exhibited particularly strong differential expression and emerged as candidate mediators of MASLD resistance. This finding was further supported by weighted gene coexpression network analysis, which identified a core regulatory network driven by the TFs HMBOX1 and PATZ1, presumably regulating 41% of its differentially expressed genes. Collectively, these results suggest that a distinct transcriptional program involving HMBOX1/PATZ1 and their downstream targets, including secreted factors, contributes to the MASLD-resistant phenotype of Göttingen minipigs and may provide promising targets for the prevention and treatment of fatty liver disease.

MCL-1 (myeloid cell leukemia-1) promotes survival and confers therapeutic resistance in acute myeloid leukemia (AML), particularly in high-risk subtypes harboring KMT2A rearrangements (KMT2A-r). Clinical trials involving patients with hematological malignancies treated with MCL-1 inhibitor monotherapy have been hampered by dose-limiting toxicity and poor response rates. Therefore, we sought to identify combinatorial treatment approaches to enhance the efficacy of MCL-1 inhibitors with the goal of improving response rates and limiting toxicities. Here, we report the inhibition of electron transport chain (ETC) complex I (CI) function as a synthetic lethal partner for MCL-1 inhibition. Co-targeting CI and MCL-1 synergistically reduces the viability in AML cell lines and patient-derived xenograft (PDX) samples in vitro, while significantly prolonging survival in mice bearing PDX AML, indicating the preclinical potential for this combinatorial therapy. These findings provide a mechanistic rationale and preclinical evidence for dual inhibition of MCL-1 and CI as a therapeutic strategy, offering a potential path to overcome resistance to single-agent MCL-1 inhibitors and improve outcomes for patients with high-risk AML. Mechanistically, we reveal that CI inhibition induces the activation of the integrated stress response, resulting in ATF4 activation downstream of the eIF2α kinase, HRI (Heme-regulated inhibitor). HRI activation via CI inhibition is dependent on the mitochondrial stress messenger, DELE1. Together, these results indicate that co-inhibition of MCL-1 and ETC CI function has the potential for improving responses in patients with KMT2A-r AML.

Mitophagy receptors are central regulators of mitochondrial quality control, integrating metabolic, stress-related, and developmental cues to maintain cellular homeostasis. Accumulating evidence indicates that their dysregulation contributes to a broad spectrum of human diseases through highly context-dependent mechanisms. In cardiovascular and neurological disorders, receptor-mediated mitophagy shapes cell survival, synaptic function, stress adaptation, and tissue integrity, with both insufficient and excessive activity proving detrimental. In cancer, mitophagy receptors display dual and stage-specific roles, acting as tumor suppressors in early disease while later supporting metabolic adaptation, stemness, and therapy resistance. Metabolic diseases highlight the tissue-specific complexity of mitophagy regulation, where precise control of mitochondrial turnover is essential for insulin sensitivity, calcium signaling, and energy homeostasis. In hematological, inflammatory, and autoimmune disorders, receptor-mediated mitophagy emerges as a fundamental determinant of lineage commitment, immune cell function, and inflammatory balance. Collectively, these findings position mitophagy receptors not as uniform stress responders, but as dynamic modulators of disease progression, whose precise and context-sensitive targeting may offer novel therapeutic opportunities across diverse pathological conditions.

Hepatocellular carcinoma (HCC) is the third leading cause of cancer death globally, often arising on a background of cirrhosis. Here, we aimed to establish genetic drivers of all-cause HCC across ancestries in a large meta-analysis. We included 15 cohorts comprising 17,697 HCC affected individuals and 2,715,683 control subjects in this meta-analysis. We found 15 genome-wide significant (p < 5 × 10^-8) germline loci, including in/near GCKR, MTTP, ADH5, 8q24.21 (nearest gene MYC), MAP3K9, and GABPB2, and a further two loci found on transcriptome- and regulome-wide association analyses. MAP3K9, TERT, and GABPB2 variants act independently of cirrhosis on both colocalization analysis and sensitivity analyses. There was significant ancestral heterogeneity in six loci including variants in the HLA locus that had divergent effects on HCC risk between East Asian and European ancestries. Fine mapping identified 11 potentially causal coding variants, including p.Leu446Pro (c.1337T>C) in GCKR and p.Asp423Glu (c.1269C>T) in MEN1. MEN1, 8q24.21 (nearest gene MYC), and TERT are all involved in the β-catenin pathway transactivation complex. Transcriptome-wide analysis identified enrichment of germline-encoded DHRS1 in HCC. Regulome-wide analysis replicated the germline signal for EPHA2 and found a chromatin-accessible region containing genes ZNF367 and HABP4. Finally, we demonstrated that population-level genetic architecture for HCC overlaps with steatotic and viral liver disease, and individuals with genetic risk for lower body mass index have higher risk of HCC. Genetic risk for HCC is determined by germline susceptibility to β-catenin pathway activation and cirrhosis. HCC is driven by both heterogeneous and homogeneous genetic factors across ancestries.

Although microRNA-22-3p (miR-22-3p) is abundantly expressed in the avian liver, its epigenetic role in lipid homeostasis remains largely uncharacterized. To elucidate its in vivo function, 14-day-old female Qingyuan Partridge chickens were intravenously injected with lentiviral vectors to establish miR-22-3p overexpression and knockdown models. Phenotypic analysis demonstrated that miR-22-3p knockdown significantly elevated hepatic triglyceride (TG) levels (p < 0.05) and drove marked steatosis, whereas its overexpression reduced TG content. Transcriptome sequencing (RNA-Seq) revealed profound metabolic remodeling, identifying 23 core lipid-associated genes (e.g., ELOVL6, FADS2, ACSBG2, and PTGIS) heavily enriched in steroid biosynthesis, fatty acid metabolism, and elongation pathways. In conclusion, miR-22-3p functions as a bidirectional epigenetic rheostat that negatively regulates hepatic lipid deposition by orchestrating a multilayered polygenic network, providing novel molecular targets for mitigating avian metabolic disorders and optimizing production traits in indigenous poultry breeds.

Osteoporosis (OP) persists as the principal contributor to the global disease burden. OP is defined by reduced bone mass and impaired bone microarchitecture, thereby weakening bone strength and elevating fracture risk. Imbalanced bone remodeling is the major pathological mechanism of OP, in which osteoclast-mediated bone resorption exceeds osteoblast-mediated bone formation. Iron serves as a vital micronutrient for numerous biochemical activities and is essential for many cellular processes. Ferroptosis is an iron-dependent form of modulated cell death with unique traits, including disrupted iron balance, compromised antioxidant defenses, and aberrant lipid peroxidation. Ferroptosis participates in various physiological and pathological processes, and its role in bone-related diseases, particularly OP, is increasingly being explored. Hence, a systematic examination of the mechanisms modulating ferroptosis in OP is imperative to pinpoint potential therapeutic targets and innovate novel therapeutic and/or preventive strategies. The agents currently used to treat OP have numerous side effects, prompting increased research on natural compounds for OP treatment. This review systematically summarizes the key features and modulation mechanisms of ferroptosis based on the latest research advances, and explores its pathogenic implications and therapeutic opportunities in OP. Additionally, we also discussed investigations of natural products <i>in vitro</i> and <i>in vivo</i> to prevent OP by interfering with ferroptosis.

The serine/threonine kinase glycogen synthase kinase-3 (GSK-3) was initially identified and studied in the regulation of glycogen synthesis. In some cases, suppression of GSK-3 activity by phosphorylation by Akt and other kinases has been associated with cancer progression. In these cases, GSK-3 has tumor suppressor functions. In other cases, GSK-3 has been associated with tumor progression by stabilizing components of the beta-catenin complex. In these situations, GSK-3 has oncogenic properties. The Raf kinase inhibitor protein (RKIP) has been reported to be under expressed in many cancers and plays a role in the regulation of tumor cells' survival, proliferation, invasion, and metastasis, hence, a tumor suppressor. RKIP also regulates tumor cell resistance to cytotoxic drugs/cells. Likewise, the tumor suppressor, phosphatase and tensin homolog (PTEN), which inhibits the phosphatidylinositol 3 kinase (PI3K)/protein kinase B (Akt) pathway, is either mutated, under expressed, or deleted in many cancers and shares with RKIP its anti-tumor properties and its regulation in resistance. Several pathways are regulated by RKIP, GSK-3, PTEN, and the transcriptional and post-transcriptional regulations of RKIP, GSK-3, and PTEN are significantly altered in cancers. In addition, RKIP, GSK-3 and PTEN play a key role in the regulation of tumor cells response to chemotherapy and immunotherapy. In this review, we will focus on the roles that GSK-3, PTEN, and RKIP play in various human cancers. We will also discuss how this pivotal kinase interacts with multiple signaling pathways such as: PI3K/PTEN/Akt/mechanistic target of rapamycin complex 1 (mTORC1), nuclear Factor kappa-B (NF-κB)/Snail family transcriptional repressor 1 (Snail)/Yin Yang 1 (YY1) loop, and rat sarcoma virus oncogene (Ras)/rapidly accelerated fibrosarcoma (Raf)/mitogen-activated protein kinase (MEK)/extracellular signal-regulated kinase (ERK).

Conditioned immune responses demonstrate that learned sensory cues can modulate peripheral immunity without re-exposure to the original immunological trigger. In 2026, this research area reaches the centennial of early Pavlovian immune-reflex experiments that already reported conditioned leukocyte shifts and enhancement-like resistance to infection. Although modern psychoneuroimmunology has been shaped largely by conditioned immunosuppression, conditioned immune enhancement remains comparatively less explored despite its relevance to anticipatory host defense, immune surveillance, tumor biology, and neuroimmune regulation. This mini review focuses on the efferent, or recall, pathways of conditioned immune enhancement and separates them from afferent acquisition signals and central cue-immune-state representations. Classical odor-conditioning paradigms using camphor and the viral mimic polyinosinic:polycytidylic acid identified interferon-<i>β</i> as an acquisition-related signal, whereas recall studies implicated β-endorphin, <i>μ</i>-opioid receptor, glutamatergic/NMDA, monoaminergic, catecholaminergic, cholinergic, serotonergic, ACTH-related, interferon-<i>α</i>-related, and endocrine mechanisms. Conditioned enhancement has been shown for natural killer cell activity, cytotoxic T-lymphocyte responses, neutrophil activity, antibody responses, and tumor-model readouts. We further integrate recent circuit-level studies of insular immune-state retrieval, brain-to-spleen humoral control, vagal cytokine coding, and descending sympathetic inflammatory pathways, together with transcriptomic data from a gene-agnostic multi-tissue pilot study assessing post-recall gene-expression dynamics. Together, these findings argue against a simple hypothalamic-pituitary-adrenal axis model and instead support a temporally organized, multi-channel efferent architecture. Dissecting and understanding this emerging architecture may provide the mechanistic basis for translating the efferent arm of conditioned immune enhancement into future therapeutic concepts with clinical impact.

<h4>Background</h4>Ferroptosis is a distinct form of regulated cell death characterized by iron-dependent lipid peroxidation, which plays crucial roles in tumor biology and therapeutic response. However, the prognostic significance of ferroptosis-related genes (FRGs) in lung adenocarcinoma (LUAD) remains incompletely understood. This study aimed to construct and validate a ferroptosis-related prognostic model for LUAD by integrating single-cell and bulk transcriptomic data.<h4>Methods</h4>Single-cell RNA sequencing (scRNA-seq) data from GSE127465 were integrated with bulk transcriptomic data from The Cancer Genome Atlas lung adenocarcinoma cohort (TCGA-LUAD). FRGs were identified through single-cell gene set scoring with the AUCell algorithm and differential expression analysis. A least absolute shrinkage and selection operator (LASSO)-Cox regression model was constructed and externally validated using the GSE72094 and GSE68465 cohorts. Associations between the model, genomic features, and the tumor immune microenvironment were systematically analyzed. The functional role of <i>SRSF9</i> was verified through <i>in vitro</i> experiments.<h4>Results</h4>A total of 93 candidate genes significantly associated with ferroptosis activity were identified, and an 8-gene prognostic signature (<i>PEBP1, TXN, TIMP1</i>, <i>SRSF9</i>, <i>CFL1, OTUB1, FURIN</i>, and PTPN6) was constructed. The signature demonstrated robust prognostic performance across validation cohorts, with time-dependent area under the curve (AUC) values ranging from 0.65 to 0.74. High-risk patients exhibited elevated tumor mutation burden (TMB) and distinct immune infiltration patterns. Consensus clustering analysis classified patients into two ferroptosis-related molecular subtypes with significantly different prognoses. Functional experiments demonstrated that <i>SRSF9</i> knockdown inhibited LUAD cell proliferation, migration, and invasion. Moreover, <i>SRSF9</i> knockdown downregulated glutathione peroxidase 4 (GPX4) and SLC7A11, increased lipid peroxidation and malondialdehyde (MDA) levels, and the resulting cell viability loss was rescued by ferrostatin-1, indicating that <i>SRSF9</i> knockdown induces ferroptosis in LUAD cells.<h4>Conclusions</h4>The ferroptosis-related prognostic signature established in this study may facilitate risk stratification in LUAD patients. <i>SRSF9</i> may regulate ferroptosis-related pathways through modulation of GPX4 and SLC7A11, and functional experiments further confirmed that <i>SRSF9</i> knockdown induces ferroptosis in LUAD cells, highlighting SRSF9 as a candidate therapeutic target that warrants further investigation.

Neurodegenerative diseases are characterized by the accumulation of misfolded and aggregation-prone proteins, reflecting a failure of proteostasis. The ubiquitin-proteasome system (UPS), a major pathway for selective intracellular protein degradation, is essential for maintaining neuronal protein homeostasis. Proteasome dysfunction has been implicated in several major neurodegenerative disorders, including Alzheimer's disease (AD), Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS), and Huntington's disease (HD), although its extent and mechanisms vary across disease contexts. In this review, we examine current evidence for proteasome dysfunction in neurodegeneration and discuss how disease-associated proteins impair proteasome function through direct inhibition, defective substrate processing, and sequestration into protein aggregates. We also address the contribution of oxidative stress, neuroinflammation, and aging to proteasome dysregulation. Finally, we highlight emerging therapeutic strategies aimed at restoring proteasome function, including pharmacological activation, modulation of proteasome assembly and stability, and targeted protein degradation approaches. Understanding the context-dependent nature of proteasome dysfunction will be important for developing effective proteostasis-based therapies.

Circulating cell-free DNA (cfDNA) enables minimally invasive assessment of chromatin organization and DNA modifications. Whether such information can be reliably recovered under conditions of limited plasma input (below 1 mL) and ultra-low sequencing depth remains unclear. We performed low-pass whole-genome Oxford Nanopore sequencing (down to 0.01× coverage) of plasma cfDNA from young and elderly donors and jointly analyzed fragment length distributions and base modifications (5mC, 5hmC, 6mA). In parallel, we analyzed an enzymatically fragmented model DNA system to assess whether controlled in vitro fragmentation can reproduce cfDNA-like nucleosomal profiles and associated modification patterns. Despite shallow coverage, cfDNA samples displayed reproducible mono-, di-, tri-, and tetra-nucleosomal peaks, indicating that major fragmentomic features can be retained under ultra-low-coverage conditions. Modification-aware basecalling enabled exploratory quantification of global modification fractions across nucleosomal size classes and nomination of candidate group-specific modification loci. Overall, these results support the feasibility of low-pass nanopore sequencing as an exploratory framework for simultaneous cfDNA fragmentomic and epigenomic profiling in low-input studies.

Treatment of inflammatory bowel disease (IBD) remains a major medical challenge due to the lack of safe and effective therapeutic agents. JAK1 has been validated as a key therapeutic target that modulates the pathological progression of IBD. In this study, using tofacitinib as the lead compound, we adopted a scaffold growth strategy to design and synthesize a series of pyrrolo [2,3-<i>b</i>] pyridine derivatives as novel JAK1 inhibitors for the treatment of IBD. Among them, compound <b>15</b> exerted potent inhibitory activity against JAK1 with an IC₅₀ value of 0.48 nM. Western blot results showed that compound <b>15</b> significantly inhibited LPS-induced STAT1/3 phosphorylation in RAW264.7 cells. In addition, <b>15</b> exhibited satisfactory metabolic stability and oral bioavailability. In the DSS-induced colitis model, <b>15</b> remarkably ameliorated inflammatory symptoms, promoted epithelial repair, and inhibited the production of pro-inflammatory cytokines such as TNF-α and IL-6. Therefore, compound <b>15</b> is regarded as a promising candidate for the treatment of IBD.

Taxanes represent a crucial class of chemotherapeutic agents clinically employed for the treatment of various malignancies, including breast cancer. Three commonly utilized taxanes-paclitaxel (Taxol), docetaxel (Taxotere), and cabazitaxel-are substantially limited by inherent drawbacks such as poor aqueous solubility, rapid clearance, and non-specific distribution. To enhance their anti-tumor efficacy against breast cancer, an mPEG-PLA-(LA)₄ nanomicelle carrier was designed and synthesized in this study for the loading of these three taxanes. These nanomicelles were designed to improve the aqueous compatibility and nanomicelle-mediated delivery of hydrophobic taxanes. In vivo animal experiments were conducted to evaluate the therapeutic effects of the three drug-loaded nanomicelles on subcutaneous human breast cancer MCF-7 xenografts in BALB/c nude mice. The results demonstrated that the injectable paclitaxel, docetaxel, and cabazitaxel micelles exhibited significant inhibitory effects on the MCF-7 xenograft tumors. These findings suggest that mPEG-PLA-(LA)₄ DS nanomicelles may serve as a structurally defined and versatile carrier platform for hydrophobic taxane delivery.

This study evaluates the chemical composition and bioactivities of essential oils extracted from the leaves and twigs of <i>Glycosmis lanceolata</i> growing in a natural forest in Vietnam. gas chromatography-mass spectrometry identified 42 and 43 constituents in the leaf and twig oils, respectively. The main compounds in the leaf oil were (<i>E</i>)-<i>β</i>-caryophyllene (10.2%), <i>β</i>-bisabolene (23.7%), and brevifolin (21.3%), while the twig oil was dominated by <i>β</i>-bisabolene (11.6%) and brevifolin (12.7%). Neither oil exhibited inhibitory effects against two beneficial bacterial strains, <i>Bacillus subtilis</i> and <i>Lactobacillus fermentum</i>. In contrast, both oils showed weak antimicrobial activity against four pathogenic bacteria-<i>Staphylococcus aureus</i>, <i>Salmonella enterica</i>, <i>Escherichia coli</i>, and <i>Pseudomonas aeruginosa</i>-and one yeast, <i>Candida albicans</i>, with IC₅₀ values ranging from 2012 ± 118 to 10,593 ± 557 µg/mL. Notably, the twig oil demonstrated pronounced anti-inflammatory activity via inhibition of nitric oxide production (IC₅₀ = 29.7 ± 2.58 µg/mL), whereas the leaf oil showed no detectable activity within the tested concentrations. Similarly, DPPH radical scavenging assays indicated stronger antioxidant activity for the twig oil compared to the leaf oil. These findings provide new insights into the phytochemistry and bioactivities of <i>G. lanceolata</i> essential oils and may support further investigations into their potential applications.

This study evaluated the association between Intimate Partner Violence (IPV) and mental health indicators, providing evidence to inform maternal health policy. This study used the first nationally representative cross-sectional data on mental health symptoms from the 2024 Zambia Demographic and Health Survey (ZDHS), including 13,951 women aged 15-49 years. Anxiety and depression were assessed using the GAD-7 and PHQ-9 scales, with a score of ≥10 indicating the presence of symptoms. Stepwise, survey-weighted multivariable logistic regression was employed to examine the associations of mental health indicators and various dimensions of IPV, reporting adjusted odds ratios (aORs) and 95% confidence intervals (CIs). Physical IPV was significantly associated with higher odds of Depression aOR= 3.89; 95% CI: 2.16-6.95; p < 0.001), while Emotional IPV was also associated with higher odds of Depression aOR = 3.50; 95% CI: 2.09-5.86; p < 0.001). Similarly, Any IPV was associated with substantial higher odds with Depression aOR = 2.90; 95% CI: 1.80-4.67, p < 0.001) and Anxiety aOR = 2.05; 95% CI: 1.20-3.50, p = 0.008). These findings provide evidence-based, actionable insights for policymakers to integrate mental health and IPV prevention in high-burden provinces to meet the targets of SDG 3 and SDG 5.

Glioblastoma (GBM) is the most aggressive primary brain tumor, and is characterized by rapid growth, therapeutic resistance, and a highly immunosuppressive tumor microenvironment (TME). Progranulin (GRN/PGRN), a secreted glycoprotein, promotes malignancy in various cancers, yet its role in GBM remains poorly defined. Bioinformatics analysis of GEO (Gene Expression Omnibus) and TCGA (The Cancer Genome Atlas) datasets revealed that GRN expression is significantly upregulated in GBM and is associated with shorter overall survival and progression-free survival. High GRN levels correlated strongly with increased M2 macrophage infiltration (marked by CD163 expression) and elevated immune and stromal scores, suggesting that GRN contributes to an immunosuppressive TME. In vitro, silencing GRN in GBM cells markedly inhibited cell proliferation, migration, invasion, and colony formation. Coculture assays demonstrated that GRN knockdown reduced GBM-induced M2 macrophage polarization (CD206⁺) while increasing M1 polarization (CD86⁺), accompanied by decreased secretion of the immunosuppressive cytokines IL-10 and TGF-β. Transwell experiments further showed that GRN promoted GBM invasion in a macrophage-dependent manner via the TGF-β pathway, an effect that was abrogated by the TGF-β receptor I inhibitor LY3200882. We developed SORT1-loaded hyaluronic acid methacrylate (HAMA) hydrogel microspheres to target GRN therapeutically. SORT1 competitively binds to GRN, reducing its concentration in the TME. Intratumoral injection of these microspheres significantly suppressed GBM xenograft growth in vivo. Collectively, our findings identify GRN as a key driver of GBM progression and TME immunosuppression through M2 macrophage polarization and TGF-β signaling. Targeting GRN with SORT1-loaded HAMA microspheres represents a promising adjuvant therapeutic strategy for GBM.

Hepcidin, a 25-amino-acid peptide hormone produced primarily by hepatocytes, is the master regulator of systemic iron homeostasis. By binding the cellular iron exporter ferroportin and inducing its internalization and lysosomal degradation, hepcidin restricts iron entry into plasma from enterocytes, macrophages, and hepatocytes. Its transcription is governed by an intricate molecular network that integrates iron status, erythropoietic demand, oxygen tension, and inflammation, with the BMP-HJV-ALK2/SMAD axis acting as the canonical activating pathway and erythroferrone (ERFE) and matriptase-2 (TMPRSS6) as physiological suppressors. Dysregulation of hepcidin underpins a wide spectrum of human diseases: insufficient hepcidin drives hereditary hemochromatosis and the iron overload of congenital and acquired ineffective erythropoiesis diseases and other ineffective erythropoiesis syndromes, whereas excessive or inappropriate hepcidin contributes to anemia of inflammation, anemia of chronic kidney disease, iron-restricted erythropoiesis in cancer, the iron-restrictive anemia of myelofibrosis, and pathogen-restrictive nutritional immunity. Within the myeloproliferative neoplasm spectrum, the divergent hepcidin patterns observed in polycythemia vera (suppressed) and myelofibrosis (inappropriately elevated through dual BMP/ACVR1/SMAD and IL-6/STAT3 hyperactivation) exemplify the clinical relevance of this axis and underpin two opposite pharmacologic strategies. Over the past decade, hepcidin pathway pharmacology has matured from proof-of-concept to regulatory milestones, shifting perspectives on several diseases and markedly improving clinical approaches.

Expansion of a disease-specific tandem repeat is responsible for >45 repeat expansion diseases (REDs). The expansion mutation in each of these diseases has different pathological consequences, and most are currently incurable. If the underlying mechanism of mutation is shared, a strategy that slows repeat expansion in one RED may be applicable to multiple REDs. However, the fact that PMS2, a component of the MutLα mismatch repair complex, promotes expansion in some models and protects against it in others suggests that the expansion mechanisms may differ. We show here using mouse models of two REDs caused by different repeats that PMS2 has similar effects in both models, with the loss of PMS2 resulting in an increase in expansions in some tissues and a loss of expansion in others. This is consistent with a protective effect of PMS2 in the first case and a role in promoting expansion in the second. Furthermore, we show in mouse embryonic stem cells that lower levels of PMS2 promote expansion while higher levels protect against it, with the ability to promote expansion depending on the PMS2 nuclease domain. Our findings lend support to the hypothesis that REDs share a common expansion mechanism and provide insights into the processes involved.

Growing evidence indicates that nuclear architecture is severely altered in many pathological contexts, primarily in cancer, with major implications for chromatin arrangement and, consequently, gene expression. Actin microfilaments located in the perinuclear region at the apical surface of cells, collectively known as the "perinuclear actin cap", integrate mechanical and biochemical cues from the cell membrane and translate them into compressive forces acting on the nuclear envelope, thereby modulating nuclear shape and size. In concert with well-established mechanotransduction paradigms, these highly dynamic and finely tuned stress fibers are emerging as key players in several biological processes - from cell migration to sensing of the surrounding microenvironment - with significant implications for development, genetic disorders and tumor progression. However, how classic pathogenetic mechanisms intersect with perinuclear actin remodeling remains mostly unknown. In this review, we will describe in detail the unique functional and structural features of perinuclear actin stress fibers and recapitulate current knowledge on their upstream regulation by membrane receptors signaling. Finally, we will explore how alterations of the perinuclear actin cap may contribute to different pathogenetic processes, with a particular focus on cancer progression and metastasis.

H5 highly pathogenic avian influenza viruses (HPAIVs) are prevalent in birds, causing infection in over 60 mammalian species worldwide. However, species-specific viral-host response at the single-cell level and within-host evolution upon H5 HPAIV infection are poorly understood. Here, by integrating spatial transcriptomics and single-cell genomics technologies, we present a comprehensive single-cell transcriptomic landscape of 1499 669 cells in the lungs from birds (chickens, ducks, and pigeons) and mammals (mice and pigs) upon H5 HPAIV infection. H5 HPAIV primarily infected avian endothelial cells and mammalian epithelial cells, respectively. Among the single-cell transcriptomic lung landscape, H5 HPAIV infection induced significant inflammation in fibroblasts and the shedding of alveolar epithelial cells of chickens upon infection, while the expression levels of cytokine- and inflammation-related genes slightly decreased in fibroblasts and endothelial cells of infected ducks and pigeons, respectively. However, mice only exhibited significantly antiviral response in macrophages and neutrophils, while antiviral response was elevated in most subtype cells of pigs upon infection. Notably, we identified 19 viral non-synonymous intrahost single nucleotide variants in the infected birds and mammals and assessed their molecular characterizations <i>in vitro</i>. Three of them were identified to increase polymerase activity and viral replication <i>in situ</i>, among which the PB2-E627K mutation was enriched in the alveolar structure of lungs, accompanied by elevated expression of interferon-stimulated genes. Collectively, we elucidated distinct viral replication, host response, and viral adaptation in the lungs across diverse birds and mammals upon H5 HPAIV infection.

Calcinosis cutis represents a debilitating complication of chronic cutaneous lupus erythematosus; however, its underlying molecular mechanisms remain poorly understood. We performed spatial transcriptomic profiling on skin biopsies from 4 patients with chronic cutaneous lupus erythematosus: 2 with calcinosis cutis and 2 without. Analysis of 87,730 cells across 6 major cell types revealed distinct molecular signatures associated with calcification. Fibroblasts in calcinosis cutis lesions exhibited significant upregulation of osteogenic and extracellular matrix genes, including periostin, collagen family members, and matrix metalloproteinase 2, accompanied by enrichment of pathways related to extracellular matrix organization and ossification. Endothelial cells demonstrated activation of hypoxia-responsive pathways, particularly hypoxia-inducible factor 2-alpha. Macrophages showed elevated chemokine receptor expression with corresponding ligand upregulation in endothelial cells, establishing a potential chemotactic recruitment axis. Spatial analysis revealed that these pathological features were concentrated in pericalcinosis regions, with reduced intercellular distances between functionally related cell populations. These findings demonstrate coordinated fibroblast osteogenic reprogramming, endothelial hypoxic responses, and immune cell activation within organized pericalcinosis niches. This spatial transcriptomic characterization of lupus-associated calcinosis cutis identifies potential therapeutic targets and reveals mechanisms resembling those in other calcifying disorders.

Transcription factors (TFs) are key drivers of tumorigenesis because of their crucial role in regulating aberrant gene expression. They contribute to tumor cell proliferation, invasion, and migration and play a pivotal role in enabling tumors to evade immune detection. In the tumor immune microenvironment (TIME), TFs reprogram tumor-infiltrating immune cells to exert pro-tumor and anti-tumor effects. In this review, we have proposed a novel, mechanism-driven classification of TFs, categorizing them into direct-acting, trans-cellular coordinated, and dual-role TFs. We have investigated the roles of direct-acting TFs in regulating CD8⁺ T cell exhaustion, maintaining CD8⁺ T cell effector functions, influencing regulatory T cell infiltration and epigenetic modifications, modulating the polarization and infiltration of tumor-associated macrophages, and promoting pro-tumor or anti-tumor properties of natural killer cells, dendritic cells and Myeloid-derived suppressor cells. In addition, we have emphasized the trans-cellular coordinated TFs that serve as bridges, facilitating cooperation among different immune cells to remodel the TIME. Finally, we have highlighted dual-role TFs that exhibit opposing functions dictated by distinct isoforms, splice variants, or post-translational modifications. Additionally, we highlight emerging pharmacological strategies targeting TFs, emphasizing their clinical potential to reverse TIME immunosuppression and synergize with immune checkpoint inhibitors. With an enhanced understanding of the molecular mechanisms underlying tumorimmune system interactions within the TIME, next-generation therapeutic strategies targeting TFs can be developed.

CAG repeat tracts occur in both non-coding and translated RNAs, have tended to lengthen throughout evolution, and are thought to enhance neuronal function. We identified over 600 human RNAs (including mRNAs, lncRNAs, and circRNAs) with at least 10 CAG repeats, originating from 58 genomic <i>loci</i>, which vary, e.g. in the rate of CAG length polymorphism. Several circRNAs originate from the <i>ATXN7</i> locus, where CAG expansion causes spinocerebellar ataxia type 7 (SCA7). For selected <i>circATXN7(3,4).1</i> (circ1), we demonstrated its cytoplasmic localization, as well as its presence in 40S, monosome and polysome fractions. We showed that circ1 is expressed in human fibroblasts, blood and cerebellum, and, importantly, we identified a mutation-containing circRNA with potential implications in SCA7.

<h4>Background</h4>HFE p.C282Y (rs1800562) homozygotes, including those with normal iron phenotypes, have increased risks of infection and death from infections, although serum IgA and IgM in cohorts of adults with p.C282Y homozygosity are not reported.<h4>Methods</h4>We compiled serum IgA and IgM levels at diagnosis of hemochromatosis in probands with p.C282Y homozygosity, investigated associations of IgA and IgM with clinical characteristics, blood count measures, and iron phenotypes, and compared mean IgA and IgM of probands with combined/weighted means of published adult European cohorts not selected for hemochromatosis.<h4>Results</h4>There were 73 probands (36 men, 37 women; mean age 51 ± 13 y). Fifty probands (68.5%) had human leukocyte antigen (HLA)-A*03. Mean IgA ± standard deviation [95% confidence interval] was 2.11 ± 1.06 g/L [1.87, 2.35]. Mean IgM was 1.11 ± 0.75 g/L [0.94, 1.28]. IgM was inversely associated with age (Pearson's r73 = -0.2733; p = 0.019). A multiple regression on IgA revealed no significant association with other characteristics. A regression on IgM revealed one positive association (daily alcohol intake; p = 0.036) and one negative association (age; p = 0.016). Mean IgA of male and female probands and corresponding mean IgA of Europeans in two cohorts (918 men, 458 women) did not differ significantly. Mean IgM of probands was lower than the mean IgM of Europeans in four cohorts (men 1.03 ± 0.84 g/L vs. 1.35 ± 0.55 g/L (n = 1084), respectively (p < 0.001); women 1.18 ± 0.67 g/L vs. 1.57 ± 0.68 g/L (n = 622), respectively (p < 0.001)).<h4>Conclusions</h4>There is no significant association of serum IgA in HFE p.C282Y homozygotes with the clinical and laboratory characteristics we studied. Serum IgM levels are positively associated with daily alcohol intake, are inversely associated with age, and are lower than those of Europeans not selected for hemochromatosis.

Adenomyosis is a common gynecological condition characterized by ectopic endometrial-like tissue deep within the myometrium, causing debilitating symptoms. Treatment options are limited to hormones that primarily focus on symptom management only or invasive hysterectomy. Despite its prevalence, the pathogenesis of adenomyosis remains poorly understood, considerably impeding the development of targeted, true disease modifying treatments. Here, through spatial transcriptomics analysis of full-thickness human uterine wall biopsies (<i>n</i> = 10), we demonstrate that adenomyosis lesions exhibit a distinct transcriptomic profile, intermediate between matched endometrium and myometrium, characterized by enhanced epithelial ciliation and altered biological pathways. We reveal that adenomyosis lesion transcriptome more closely resembles the endometrial basalis than the functionalis, marked by enriched olfactory signaling, dysregulated oxidative phosphorylation and adenosine triphosphate synthesis, and an altered immune microenvironment indicative of chronic inflammation. In silico drug screening identified candidate compounds capable of reversing the adenomyosis lesion transcriptomic signature. These findings offer previously unknown insights into the molecular landscape of adenomyosis lesions and lay the groundwork for development of targeted, lesion-specific therapies that may preserve the eutopic endometrium and offer alternatives to hysterectomy.

Domestic dogs exhibit substantial morphological diversity, making quantitative characterization of their phenotypes challenging. Traditional phenotyping methods often rely on manual measurements, which are limited in their ability to capture complex visual traits. Deep learning provides an opportunity to automatically extract informative and biologically meaningful features from images. In this study, we constructed a dataset of 13,254 dog images across multiple breeds and used ResNet and ViT models to automatically extract 256-dimensional image embeddings. After dimensionality reduction using UMAP (uniform manifold approximation and projection), we performed a GWAS (genome-wide association study) on the extracted features and breed-level genotype data. We identified 15 genes previously reported to be associated with dog traits such as hair length and body size, as well as previously unknown candidate genes related to body development and hair growth, including <i>EIF2S2</i>, <i>TRHR</i>, and <i>TCF25</i>, which harbor variants with potential functional relevance. This approach is validated by known genetic associations and can reveal previously unidentified genotype-phenotype links. Building on these capabilities, this approach provides a scalable framework for phenotype extraction that enables population genetic studies in domestic dogs and can facilitate breeding in other economically important species.

<h4>Background</h4>Major depressive disorder (MDD) exhibits a higher prevalence in women, yet the underlying cellular mechanisms for this clinical disparity remain elusive. To investigate sex-specific molecular mechanisms, we profiled sex-specific molecular responses to sub-chronic stress in the mouse hippocampus at single-nucleus resolution.<h4>Methods</h4>Male and female C57BL/6N mice were exposed to sub-chronic variable stress (SCVS). Anhedonia was assessed via sucrose preference and novelty-suppressed feeding. We employed single-nucleus RNA sequencing (snRNA-seq) to map sex-specific hippocampal responses to sub-chronic variable stress (SCVS) in mice.<h4>Results</h4>Female mice displayed significantly exacerbated anhedonia and selective depletion of plasma serotonin (5-HT) following SCVS exposure, contrasting with minimal behavioral changes in males. Single-nucleus RNA sequencing of 31,256 hippocampal cells revealed profound sex-dimorphic transcriptional reprogramming: females exhibited a 3-fold greater number of differentially expressed genes (DEGs) than males with minimal overlap. Key mechanistic findings included: (1) Females exhibited oligodendrocyte-specific upregulation of mitochondrial genes (mt-Atp6, mt-Co3), increasing oxidative stress susceptibility. Concurrently, stress depleted their baseline oligodendrocyte predominance, potentially disrupting hippocampal synchrony underlying anhedonia.; (2) Females exhibit broad Mef2c upregulation, while males show astrocyte/cholinergic neuron NRG-1 upregulation and astrocyte Slc6a11 downregulation; (3) Females display suppressed oxytocin signaling and coordinated dampening of glutamatergic signaling, astrocytic K⁺ buffering, and cell adhesion; males exhibit dysregulated kinase/phosphatase activity.<h4>Conclusion</h4>This study presents the single-nucleus atlas of hippocampal sexual dimorphism in stress response, identifying oligodendrocyte mitochondrial dysfunction, pan-cellular Mef2c upregulation, and oxytocin suppression as potential female-specific therapeutic targets for depression. Our findings reveal fundamental sex-divergent molecular adaptations to stress, providing a roadmap for novel, sex-stratified treatments for MDD.

Astrocytes coordinate neuronal activity with vascular development and metabolic homeostasis during postnatal brain maturation, yet the mechanisms enabling astrocytes to support angiogenesis remain elusive. VPS35, a core component of the retromer complex, regulates transmembrane protein trafficking and is implicated in multiple neurodegenerative diseases; however, its function in astrocytes is unknown. Here, we identify astrocytic VPS35 as a crucial regulator for astrocyte maturation and neurovascular development through its control of iron homeostasis and HIF1α-VEGFa signaling. Conditional deletion of Vps35 in astrocytes impaired astrocyte maturation and survival, as well as basal and activity-dependent angiogenesis, thus reducing cerebral blood flow, and inducing widespread vascular abnormalities in both cortical and meningeal vessels. Mechanistically, loss of astrocytic Vps35 resulted in intracellular iron accumulation, likely due to impaired ferroportin (FPN)-mediated iron export. Iron overload enhanced HIF1α hydroxylation and degradation, suppressing VEGFa expression and attenuating angiogenic responses. In parallel, excess iron induced astrocyte ferroptosis, characterized by increased lipid peroxidation, diminished antioxidant capacity, astrocyte loss, and impaired astrocyte maturation. Importantly, iron chelation reduced ferroptotic stress, restored HIF1α-VEGFa signaling, and rescued vascular density in mutant mice. Together, these findings establish astrocytic VPS35 as a critical integrator of endosomal protein trafficking, iron metabolism, and angiogenic signaling, and identify iron dysregulation as a central mechanism linking retromer dysfunction to impaired astrocyte and neurovascular development.

<h4>Background</h4>Primary biliary cholangitis (PBC) management remains limited by reliance on static biochemical markers, fragmented assessment of symptom burden, and inadequate noninvasive risk stratification for clinically significant portal hypertension (CSPH). Existing tools fail to integrate longitudinal laboratory trends, elastography, and patient-reported outcomes, and translation of risk assessment into guideline-concordant clinical action remains inconsistent.<h4>Objective</h4>This study aims to develop, validate, and pilot an explainable artificial intelligence (AI) framework-AIm-PBC-for continuous disease monitoring, early prediction of CSPH complications, and delivery of guideline-based clinical decision support in patients with PBC.<h4>Methods</h4>We will conduct a multiphase study combining retrospective cohort analysis, prospective observational data collection, and implementation evaluation. At least 600 adults with confirmed PBC will be enrolled across academic hepatology centers. The AI framework will integrate longitudinal biochemical markers, noninvasive elastography metrics, and high-frequency patient-reported outcomes to generate an interpretable disease activity index and predict CSPH-related complications using gradient-boosted models with Shapley additive explanations. Outputs will be deployed via a SMART-on-FHIR (Substitutable Medical Applications, Reusable Technologies on Fast Healthcare Interoperability Resources)-enabled electronic health record clinical decision support tool. Implementation will be evaluated using a randomized crossover simulation followed by a pragmatic pilot assessing usability, cognitive load, clinician adherence, and feasibility.<h4>Results</h4>The primary outcomes include calibration and responsiveness of the disease activity index; discriminatory performance of the CSPH prediction model compared with established criteria; and effectiveness of the decision support tool measured via improvements in guideline-concordant care, usability scores, and clinician cognitive workload. Secondary outcomes include fairness metrics and workflow efficiency.<h4>Conclusions</h4>This protocol outlines a scalable, explainable AI framework designed to bridge gaps between disease monitoring, risk prediction, and clinical action in PBC. If successful, AIm-PBC may enhance early complication detection, improve symptom management, and support equitable, evidence-based care delivery in routine hepatology practice.

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder characterized by progressive motor neuron loss. In addition to the different pathogenic mechanisms, in recent years, increasing attention has been directed toward the role of lipid metabolism in ALS pathogenesis, although the clinical relevance of lipid alterations in ALS may differ from their well-established role in cardiovascular disease. This review critically examines the multifactorial relationship between cholesterol and ALS through three perspectives: (1) as a risk factor for disease onset, (2) as a prognostic biomarker of disease progression, and (3) as a potential therapeutic target. Epidemiological and genetic studies suggest a complex and sometimes contradictory association between lipid profile and ALS risk. Elevated LDL-cholesterol and total cholesterol have been linked to increased disease susceptibility in some cohorts, with Mendelian randomization studies supporting a potential causal role. Conversely, evidence regarding HDL-cholesterol remains conflicting and may be influenced by sex-specific and metabolic factors. As a prognostic biomarker, hyperlipidemia has been variably associated with prolonged survival in ALS patients; however, these findings often lose significance after adjusting for body mass index and nutritional status, suggesting that lipid levels may reflect systemic metabolic reserve rather than directly modulating disease progression. Pharmacological modulation of cholesterol reveals further complexity. While statins are generally not associated with increased ALS risk in clinical studies, preclinical models show divergent effects: some statins accelerate disease progression, while others like lovastatin may be protective. Other lipid-lowering drugs, including fibrates and PCSK9 inhibitors, may also influence ALS-related pathways beyond cholesterol lowering, although their potential role remains to be clarified.

Rare neurological diseases (RND) represent a growing but underrecognized global health burden, particularly in aging populations in whom clinical manifestations appear later in life, resulting in substantial morbidity, reduced quality of life, and increased mortality. Advances in understanding and treating these diseases have been hindered by low prevalence, phenotypic heterogeneity, and complex molecular mechanisms. Importantly, shared genetic and mechanistic links between rare and common neurodegenerative disorders, such as <i>GBA1 (Glucosylceramidase Beta 1)</i> gene variant-associated Gaucher disease (GD) and Parkinson's disease (PD), highlight convergent biological pathways that may be leveraged for biomarker discovery and therapeutic innovation. Notably, individuals heterozygous for pathogenic <i>GBA1</i> variants, historically considered asymptomatic carriers, are now recognized to have an increased lifetime risk of developing PD and related synucleinopathies. This emerging evidence indicates that even single-allele variants can confer long-term neurological risk, reinforcing the continuum between rare monogenic disorders and more common neurodegenerative diseases. Extracellular Vesicles (EVs) have emerged as a promising, minimally invasive platform for advancing RND research, although translation remains limited by source specificity, vesicle heterogeneity, and incomplete clinical validation. By encapsulating proteins, lipids, and nucleic acids that reflect their cellular origin and disease state, EVs offer unique opportunities for early diagnosis, disease stratification, longitudinal monitoring, and assessment of treatment responses. In this review, we summarize fundamental aspects of EV biology and critically evaluate recent advances in EV-based biomarker discovery for RND, informed by translational insights from more prevalent neurodegenerative conditions. We further discuss the therapeutic potential of EVs, emphasizing their intrinsic biocompatibility, their reported capacity in selected preclinical settings to traverse biological barriers, including the blood-brain barrier, and their versatility as carriers for neuroprotective and gene-modifying cargo. Finally, we address key technical and translational challenges, including isolation, characterization, scalability, and regulatory considerations that currently limit clinical adoption. Collectively, this review highlights emerging opportunities and outlines an integrative translational perspective for EV-based diagnostic and therapeutic strategies into research and clinical paradigms for aging individuals affected by RND.

Post-COVID condition is a heterogeneous, multi-system sequela of SARS-CoV-2 infection that imposes substantial socioeconomic burden and currently needs validated diagnostic biomarkers or established therapeutic pathways. This brief communication synthesises blood-based proteomic and targeted biomarker evidence and organises it into three overlapping pathophysiological domains: persistent immune dysregulation (IL-6, IL-20, MCP-1 and TNF- α), endothelial dysfunction and disordered haemostasis (VEGF-A, P-selectin, vWF, ICAM-1 and D-dimer); and neurological injury (NFL, GFAP, NAAA, LXN, NBL1, HAGH). Across studies, no single protein provides adequate diagnostic performance; phenotype-linked multi-analyte panels show the greatest promise. Researching post-COVID conditions requires harmonised case definitions, cross-platform validation, and the integration of coagulation assays and extracellular vesicle profiling to improve tissue signal detection.

Skeletal diseases impose a heavy global burden, creating an urgent demand for safe and effective therapeutic strategies for bone regeneration. Quercetin, a natural flavonoid compound, has attracted considerable attention owing to its potential to promote bone formation and inhibit bone resorption. Nevertheless, its poor solubility and low bioavailability limit its clinical application. The integration of drug delivery systems with bone tissue engineering offers a promising approach to address these challenges. This review examines the latest advances in quercetin-based drug delivery systems and their synergistic mechanisms with bone tissue engineering. It systematically discusses various strategies, including hydrogels, hydroxyapatite, biofunctional scaffolds, phospholipid carriers, metal complexes, nanomaterials, and medicinal chemical modifications, and presents the core advantages of these approaches in achieving targeted delivery, controlled release, and enhanced bioactivity of quercetin. Furthermore, this review highlights the central role of these delivery systems in potentiating the pro-regenerative effects of quercetin. These advances not only provide practical solutions to overcome the inherent limitations of quercetin but also significantly broaden its application prospects in bone tissue engineering.

Diabetic kidney disease (DKD) is the leading cause of chronic kidney disease and end-stage kidney failure worldwide, yet the molecular mechanisms driving progressive renal injury remain incompletely understood. Chronic metabolic stress in diabetes disrupts endoplasmic reticulum (ER) homeostasis, leading to sustained activation of the unfolded protein response (UPR). While transient UPR signaling is adaptive and restores proteostasis, persistent ER stress in the diabetic milieu shifts UPR signaling toward maladaptive pathways that promote cellular dysfunction and death. This review summarizes the canonical UPR branches mediated by protein kinase RNA-like ER kinase (PERK), inositol-requiring enzyme 1α (IRE1α), and activating transcription factor 6 (ATF6), and discusses their dysregulation in the diabetic kidney. We highlight how chronic glucolipotoxicity, oxidative stress, and protein overload drive prolonged UPR activation in podocytes and tubular epithelial cells, leading to loss of proteostatic balance and progressive nephron injury. Importantly, emerging evidence indicates that UPR signaling interacts with multiple regulated cell death pathways, including apoptosis, autophagy dysfunction, ferroptosis, pyroptosis, and necroptosis, forming a pathological crosstalk network that determines renal cell fate. This maladaptive integration amplifies inflammation, oxidative stress, mitochondrial dysfunction, and fibrotic remodeling, ultimately contributing to podocyte depletion, tubular atrophy, and disease progression. Finally, we discuss therapeutic strategies to restore ER proteostasis and modulate UPR-mediated cell death signaling, highlighting their potential as disease-modifying approaches in DKD. A deeper understanding of UPR dysregulation and its interaction with cell death pathways may provide novel mechanistic insights and facilitate the development of targeted therapies for diabetic kidney disease.

The incidence of neurodegenerative diseases, including Alzheimer's disease (AD), continues to increase with the extension of human lifespan. However, their pathogenesis remains incompletely understood. Altered energy metabolism, particularly glucose metabolism involving glycolysis and oxidative phosphorylation, is widely recognized as an early pathological feature of neurodegenerative diseases. Astrocytes, the most numerous and widely distributed functional cells in the central nervous system (CNS), support neuronal energy demands through the astrocyte-neuronal lactate shuttle (ANLS). Glycolysis is a major pathway of astrocyte energy metabolism, and enhanced astrocytic glucose uptake and glycolytic flux may help attenuate the progression of neurodegenerative diseases such as AD. Zinc Finger and BTB Domain Containing 7A (ZBTB7A) is a POZ/BTB and Krüppel (POK) family transcription factor that has been implicated in the regulation of metabolic genes, including glycolysis-related genes, in several cellular contexts. However, its role in astrocyte glycolytic regulation under neurodegenerative conditions remains unclear. In this review, we summarize current knowledge of ZBTB7A biology, astrocyte glycolysis, and glial metabolic dysfunction in neurodegenerative diseases, and integrate published evidence with bioinformatics-based transcription factor binding prediction. Our analysis identified putative ZBTB7A-binding motifs in promoter regions of genes involved in glucose uptake, glycolytic flux, lactate production, and lactate transport. These findings suggest a potential association between ZBTB7A and the astrocytic glycolytic/lactate metabolic network. Therefore, this review provides a conceptual basis for future studies on ZBTB7A-associated transcriptional regulation in astrocyte metabolic remodeling and its potential relevance to neurodegenerative diseases.

European sea bass (<i>Dicentrarchus labrax</i>) is a seasonal marine teleost of high aquaculture value, yet a comprehensive, stage-resolved molecular description of its spermatogenic cycle has been lacking. Here, we generated an integrated histology-anchored RNA sequencing atlas covering the six canonical testicular stages (Stages I-VI) to reconstruct the temporal sequence of endocrine, proliferative, meiotic, spermiogenic and regression-related processes across the annual reproductive cycle. Early stages (Stages I-III) were characterised by strong activation of spermatogonial maintenance pathways, hormone-responsive programmes, and circadian regulators, along with robust enrichment of DNA replication, kinetochore assembly, and chromosomal segregation mechanisms that prepare germ cells for meiotic entry. Spermiogenesis (Stage IV) represented the major transcriptional shift, with the coordinated upregulation of axonemal motors, intraflagellar transport and BBSome components, outer dense fibres and microtubule regulators, accompanied by extensive chromatin remodelling involving histone variants, transcription factors, ubiquitin/SUMO pathway enzymes, and heterochromatin-associated proteins. Spermiation (Stage V) showed an enrichment in calcium-dependent signalling, cytokine and chemokine networks, and adhesion and extracellular matrix pathways, consistent with Sertoli-germ cell remodelling and sperm release. Regression (Stage VI) was dominated by immune activation, endocytosis, phagocytosis, and vascular remodelling signatures, indicating an active phase of clearance and tissue reorganisation that preserves the spermatogonial reservoir for the next cycle. From 9,327 differentially expressed genes, we identified 132 distinct temporal expression patterns and curated three translational resources: stage-informative markers, a fertility gene set, and a chromatin-associated compendium. Together, this atlas defines the sequential molecular architecture of spermatogenesis in a seasonal marine fish, conserved vertebrate and species-specific regulatory modules, and provides operational biomarkers for broodstock assessment and the mitigation of precocious maturation in aquaculture.

<h4>Objective</h4>The genetic architecture underlying susceptibility to neonatal hyperbilirubinemia in specific subtropical populations remains poorly defined. This study aimed to evaluate the association between polymorphisms in the uridine diphosphate glucuronosyltransferase 1A1(UGT1A1) and solute carrier organic anion transporter family member 1B1 (SLCO1B1) genes and the risk of neonatal hyperbilirubinemia within the Haikou population, and to explore potential cumulative genetic effects.<h4>Methods</h4>A retrospective case-control study was conducted involving 210 neonates diagnosed with hyperbilirubinemia (case group) and 195 healthy neonates (control group) admitted to the Haikou Maternal and Child Health Hospital between January 2024 and January 2025. Serum levels of total bilirubin (TBIL), indirect bilirubin (IBIL),and alkaline phosphatase (ALP) were quantified. Seven target single nucleotide polymorphisms (SNPs) were genotyped, including four UGT1A1 loci (rs4148323, rs3771341, rs34946978, rs35350960) and three SLCO1B1 loci (rs4149056, rs2306283, rs4149015). Genotype-phenotype correlations, disease risk associations,gene-gene interactions, cumulative risk allele analyses, and combined genotype analyses were statistically evaluated. Stratified and sensitivity analyses were performed to assess the robustness of the findings.<h4>Results</h4>The case group exhibited a significantly higher prevalence of preterm births compared to the control group (16.9% vs. 9.5%, <i>P</i> = 0.04). Allele frequencies for UGT1A1 rs4148323 (22.14% vs. 11.79%), SLCO1B1 rs4149056 (19.05% vs. 10.26%), and rs4149015 (21.43% vs. 11.79%) were significantly elevated in the case group (<i>P</i> < 0.05). Possession of the UGT1A1 rs4148323 GA genotype (OR = 2.370, 95% CI: 1.499-3.805), SLCO1B1 rs4149056 TC genotype (OR = 1.594, 95% CI: 1.006-2.545), and rs4149015 AG genotype (OR = 2.247, 95% CI: 1.429-3.576) was associated with a significantly increased risk of hyperbilirubinemia. In multivariate analysis adjusting for gestational age, birth weight, and feeding modality, these associations remained robust (adjusted ORs: 2.338, 1.481, and 2.233, respectively; all <i>P</i> < 0.05). No statistically significant multiplicative interaction was detected between UGT1A1 and SLCO1B1 variants (P-interaction > 0.05). However, a cumulative risk allele analysis demonstrated a significant dose-response relationship: neonates carrying 3-4 risk alleles across UGT1A1 rs4148323and SLCO1B1 rs4149056/rs4149015 exhibited a markedly elevated risk compared to those with 0-1 risk alleles (adjusted OR = 3.412, 95% CI: 1.892-6.154, <i>P</i> < 0.001). Combined genotype analysis further revealed that carriers of concurrent UGT1A1 rs4148323 variant(GA/AA) and SLCO1B1 rs4149056/rs4149015 variant (TC + CC/AG + AA) genotypes had significantly higher TBIL levels than carriers of single-gene variants (<i>P</i> < 0.01), consistent with an additive genetic burden. The primary stratified analysis restricted to term neonates (>=37 weeks) confirmed that the main genetic associations remained robust; late-preterm (35-36 weeks) subgroup analyses were underpowered and are presented in Supplementary Materials.<h4>Conclusion</h4>Neonatal hyperbilirubinemia is a multifactorial condition; our findings underscore that UGT1A1 and SLCO1B1 variants-specifically rs4148323, rs4149056, and rs4149015-are potent genetic biomarkers for predicting disease susceptibility in the Haikou population. The significant additive cumulative effect supports the clinical value of multi-locus genetic profiling. Future large-scale studies incorporating multi-gene panels and formal additive burden testing are warranted to elucidate complex gene-gene relationships and facilitate personalized neonatal care.

Inflammatory bowel diseases (IBD) are increasingly recognized as disorders in which epithelial dysfunction and maladaptive regeneration may be as important as immune dysregulation. Tumor necrosis factor (TNF), a key mediator of intestinal inflammation and a therapeutic target, plays a dual role in both immune activation and epithelial repair by regulating progenitor cell expansion, lineage plasticity, and chemokine signaling in the intestinal epithelium. During acute injury, TNF-associated responses are generally considered adaptive, supporting crypt repair, barrier restitution, and secretory remodeling pathways. However, in chronic disease, persistent TNF exposure, potentially reinforced by type I interferons (IFN-I), may contribute to the persistence of epithelial regenerative pathways. IFN-I signaling has been suggested in experimental and translational studies to reinforce chemokine networks and transcriptional imprinting. We propose that this potentially converts physiological repair into a sustained state of what we have termed "regenerative inflammation," in which epithelial-derived signals may perpetuate immune recruitment and tissue remodeling. Such TNF-IFN-imprinted epithelial states may contribute to sustained pathology in a subset of patients and could be associated with reduced responsiveness to anti-TNF therapy, although direct causal evidence in human disease remains limited. By integrating mechanistic, organoid-based, and clinical observational evidence, we propose that chronic TNF-IFN crosstalk may contribute to a self-sustaining regenerative inflammatory circuit, providing a conceptual framework for disease persistence in IBD and highlighting potential opportunities to target epithelial-immune interactions.

Targets of RNA-binding proteins (RBPs) are often investigated by implementing variants of cross-linking and immunoprecipitation methodology, which can yield several disadvantages in target detection. The RBP and <i>N</i>6-methyladenosine (m6A) reader insulin-like growth factor 2 mRNA binding protein 2 (IGF2BP2/IMP2) exerts an essential pathophysiological role as a metabolic regulator and tumor promoter, impacting the stability, localization, and translation of its targets. Here, we employed HyperTRIBE as a method to identify RBP targets in native cells <i>in vivo</i> and identified targets of IMP2 in murine hepatocytes. IMP2-associated adenosine-to-inosine editing sites were identified by hydrodynamic transfection of mouse livers using an IMP2-ADAR (adenosine deaminase acting on RNA) construct. Functional enrichment and motif analysis results suggest IMP2-facilitated target stabilization and confirm presence of m6A-binding motifs. In addition, the overlap with data of a TRIBE experiment employing murine embryonic fibroblasts and with those of differential gene expression was investigated. Comparative transcriptomics between IMP2, wild-type, and control samples (mCherry-ADAR) revealed an enrichment of IMP2-bound mRNAs associated with autophagy, which could be validated by RNA immunoprecipitation in a human liver cancer cell line. A functional knockdown of IMP2 demonstrated an increased autophagic flux, providing further evidence for the involvement of IMP2 in autophagy.

The development of biocompatible materials has gained traction due to the increasing clinical demands for customizable and functional medical devices. Chitosan, a deacetylated derivative of chitin, is a naturally occurring biopolymer with strong antimicrobial properties, immunocompatibility, and structural adaptability, making it a promising candidate for biomedical applications. Through mechanisms such as crosslinking, ionic bonding, gas formation, and UV radiation, the mechanical properties and stimulus responses of chitosan-based hydrogels can be tailored for drug delivery at specific sites or under specific pH, light, or electrical conditions. Beyond drug delivery, chitosan hydrogels have shown considerable potential for vascular tissue repair. The porous structure of chitosan allows patient specific vascular scaffolding to be created that promotes the recovery rate veins and stenting procedures. Thermally sensitive hydrogels can deliver drugs to target regions to further assist in vascular healing. Furthermore, recent developments with composite polymers and coatings engineered to self-assemble within veins provide scaffolds for vascular tissue growth. This manuscript reviews chitosan hydrogel fabrication methods and their corresponding materials properties, with particular emphasis on drug delivery to vascular tissues. Furthermore, relevant findings from clinical trials are summarized to support the potential of chitosan hydrogels for future clinical use. Challenges of chitosan hydrogels, such as insufficient mechanical strength, high degradation rates, and complex manufacturing, remain as areas for research break-through.

The most persistent biomedical challenges of the 21st century are neurodegenerative disorders (NDs), where molecular alterations lead to devastating clinical consequences and progressive neuronal loss. The prevalence of neurodegeneration is continuously rising and becoming the main contributor to chronic disability and mortality. Despite their clinical differences, many conditions share pathogenic processes, including oxidative stress, protein misfolding and aggregation, mitochondrial dysfunction, and neuroinflammation. Instead of functioning independently, these processes cooperate to form a self-reinforcing network that gradually weakens synapses and ultimately leads to neuronal death. This study redefines neurodegeneration as a disorder of system-level failure by emphasizing poor cellular stress integration. In addition to demonstrating how gut microbiome gene networks impact inflammation and amyloid production, new research highlights the relationships between mitochondrial-lysosomal interactions, endoplasmic reticulum stress responses, and transcriptionally driven synaptic vulnerability. A key molecular topic is the interaction and pathogenic convergence of the JAK/STAT, HIF-1α, and Notch signaling pathways. Under ongoing metabolic stress, prolonged stimulation of this triad increases inflammation, hinders the regenerative processes, and maintains pseudo-hypoxic conditions, explaining why single-target treatments have mostly been unsuccessful. This review also explores progress in fluid, digital, and imaging biomarkers that facilitate early diagnosis and patient stratification, and assesses new disease-modifying approaches such as antisense oligonucleotides, immunomodulators, gene therapies, and small-molecular agents. Artificial intelligence is emphasized as an essential tool for integrating multimodal data, drug discovery and predictive modeling.

<h4>Summary paragraph</h4> Highly heritable, polygenic, and easily measured, adult height has long been the model trait in human genetics 1,2 . While the landscape of height-associated common genetic variation has been studied extensively 2 , rare variation remains relatively unexplored 1 . Using rare protein-altering variants in a discovery set of 826,066 exomes, we identify 207 height-associated genes - 98% of which replicate in an additional 624,567 individuals. The rarest and most deleterious class of variation, singleton (frequency <0.0001%) putative loss-of-function (pLoF) variants implicated 17 genes with large effects on height ranging from -17 cm ( ACAN ) to +11 cm ( FBN1 ) per allele, 52× larger than the average effect of common height-associated variants and comparable to the 1% tails of a common variant polygenic score. Several genes (e.g., TET1 , DTL , IGF2BP2 ) have effect sizes at least as large as established Mendelian height genes but lack documented stature or skeletal growth syndromes. This is particularly true for genes in which rare variants associate with increased height. We performed the largest rare-variant study of height to date, directly implicate 207 genes that broadly overlap with both GWAS associations and Mendelian height syndromes, assess the impact of rare variants on heritability and prediction, provide evidence that height is an underappreciated clinical feature of Mendelian disorders, and demonstrate the utility of large population-scale sequencing studies for classifying individual variants and dissecting complex trait architecture.

Challenges in the precise diagnosis and treatment of nasopharyngeal carcinoma (NPC) remain, mainly due to the absence of a multi-omics-based molecular classification and effective targeted therapies. In this study, we performed proteomic and phosphoproteomic analyses of NPC and non-cancerous nasopharyngeal tissues to identify key dysregulated proteins and phosphorylation networks. Based on these profiles, we classified NPC into two distinct molecular subtypes: S1 and S2, which exhibit significant clinical heterogeneity. Notably, the S2 subtype displayed stronger immune suppressive characteristics. By leveraging proteomic data from both cancerous and non-cancerous tissues, as well as from the two molecular subtypes, we developed robust diagnostic and prognostic models. Through computational drug repurposing and experimental validation, we identified Panobinostat, a pan-histone deacetylase inhibitor, as a potent anti-tumor agent for NPC, demonstrating efficacy in both in vitro and in vivo models. Mechanistically, Panobinostat inhibits MYC expression, thereby suppressing the transcriptional activation of key components in the homologous recombination (HR) DNA repair pathway. This reduction in transcriptional activation impairs HR repair efficiency and leads to the accumulation of DNA double-strand breaks (DSBs). Furthermore, combination therapy with Panobinostat and radiotherapy produced a synergistic effect, significantly enhancing NPC suppression. Additionally, we predicted and validated potential drugs for targeting the S2 subtype of NPC. In conclusion, we identified molecular subtypes of NPC, constructed preliminary diagnostic and prognostic marker panels, and observed the therapeutic potential of Panobinostat, as a monotherapy and in combination with radiotherapy. These findings provide a solid foundation for precision diagnosis, prognostic stratification, and personalized treatment strategies for NPC.

Perivascular epithelioid cell neoplasms (PEComas) are ultra-rare mesenchymal tumors lacking a molecular classification to guide therapy. Here we perform comprehensive multi-omic profiling of an unselected PEComa cohort. We identify frequent MITF rearrangements involving actin gene partners (ACTA2, ACTG1 and ACTB). Anatomical stratification reveals cyclin-dependent kinase module mutations in gynecologic tumors, whereas soft tissue, gastrointestinal, and pelvic tumors lacked mTOR pathway alterations but are enriched for TFE3/MITF rearrangements. Transcriptomic analysis defines four subtypes with distinct lineage programs-melanocytic, mesenchymal, or adipogenic-as well as unique mutational patterns and clinical behaviors. Notably, an aggressive stem-like subtype enriched for TP53/RB1 mutations exhibits high proliferation, activation of embryonic and Hedgehog signaling, immune infiltration, and resistance to mTOR inhibitors, but potential responsiveness to immunotherapy. Single-nucleus RNA sequencing reveals intra-tumoral heterogeneity within this subtype, including divergent inflammatory states. Together, these findings establish a molecular classification framework and identify actionable vulnerabilities in PEComa.

Systemic lupus erythematosus (SLE) is a chronic autoimmune disease with diverse manifestations, including rash, arthritis, and nephritis. Although autoantibodies are a key feature of SLE, their levels often poorly reflect disease severity, suggesting the involvement of additional contributing factors. The tripartite motif-containing protein (TRIM) family, which regulates immunity, may play a role in SLE. Although TRIM21 is a known autoantigen, the roles of other TRIM family members remain unclear. The present study examined TRIM expression in SLE and evaluated the potential of TRIM proteins as biomarkers. TRIM expression in peripheral blood mononuclear cells was measured using reverse transcription quantitative polymerase chain reaction (PCR) and compared between patients with SLE (n = 33) and healthy controls (n = 20). Associations of TRIM expression with disease activity (active/inactive), renal involvement (nephritis/nonnephritis), clinical manifestations, and laboratory parameters were analyzed. The expression levels of several TRIM genes, specifically TRIM5, TRIM11, TRIM21, and TRIM72, were higher in patients with inactive SLE, nonnephritic disease, or low disease activity than in patients with active disease, nephritic disease, or relapsed SLE and healthy controls. The expression levels of these genes were considerably and negatively associated with clinical symptoms but not laboratory parameters. Hydroxychloroquine users exhibited higher levels of TRIM21 expression than did nonusers. Overall, specific TRIM proteins are inversely correlated with SLE disease activity and clinical phenotypes, which indicates their potential as biomarkers. Hydroxychloroquine may regulate TRIM21 expression. Further investigation into underlying mechanisms and continued TRIM monitoring are necessary to develop new SLE treatments and better assess disease progression.

Sepsis is associated with a pronounced but poorly understood endoplasmic reticulum stress (ERS) response. In this study, we found that death-associated protein kinase 2 (DAPK2), a calcium/calmodulin-regulated serine/threonine kinase, exhibits elevated expression in macrophages from patients with sepsis and from septic mice. Macrophage DAPK2 expression is transcriptionally upregulated through the activation of the Toll-like receptor 4 (TLR4)-myeloid differentiation primary response 88 (MyD88)-nuclear factor-κB (NF-κB) pathway. Macrophage-specific deletion of DAPK2 attenuated sepsis severity and mitigated the ERS response. Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we identified heat shock protein family A member 5 (HSPA5) as a binding partner for DAPK2. Because DAPK2 function had been previously associated with the kinase activity, we speculated that it might control ERS of macrophages through HSPA5 phosphorylation. Further investigation indeed revealed that DAPK2 phosphorylates HSPA5 at serine-588, which promotes the proteasomal degradation of HSPA5 and subsequently leads to the activation of inositol-requiring enzyme 1α (IRE1α). Inhibition of HSPA5 exacerbated sepsis in mice with macrophage-specific DAPK2 deficiency; however, this effect was abrogated by the deactivation of IRE1α. In conclusion, our findings demonstrate that DAPK2 propagates macrophage ERS through the HSPA5-IRE1α axis during systemic infection, suggesting this pathway as a potential therapeutic target in sepsis.

<h4>Objective</h4>To compare the effectiveness in US practice of normothermic regional perfusion (NRP) and normothermic machine perfusion (NMP) for donation after circulatory death (DCD) liver transplantation (LT).<h4>Background</h4>DCD livers historically conferred inferior outcomes to donation after brain death donors, primarily due to ischemic biliary injury. NRP and NMP have emerged as promising strategies to mitigate this risk.<h4>Methods</h4>Using UNOS data (2022-2024), we stratified DCD donors by procurement [super-rapid recovery (SRR) or NRP] and preservation [static cold storage (SCS) or NMP] method. NMP cases were further stratified into on-site versus back-to-base initiation of perfusion. On-site NMP was directly captured in the UNOS dataset. NRP and back-to-base NMP were identified with surrogate markers. We then compared graft (GS) and overall (OS) survival in propensity-matched cohorts.<h4>Results</h4>We identified 4632 DCD LTs. The most common procurement-preservation strategy was SRR-NMP (2637; 56.9%), followed by SRR-SCS (937; 20.2%), NRP-SCS (560; 12.1%), and NRP-NMP (498; 10.8%). SRR-NMP demonstrated superior GS (HR: 0.54, 95% CI: 0.32-0.92) versus SRR-SCS. NRP-SCS similarly reduced the risk of graft loss (HR: 0.42, 95% CI: 0.29-0.61) versus SRR-SCS. NRP-SCS and SRR-NMP had similar GS. Similarly, adding NMP to livers procured with NRP did not lead to improved outcomes compared with NRP followed by SCS.<h4>Conclusions</h4>This study provides the most comprehensive US comparison of modern procurement and preservation strategies in DCD LT. Both NRP and NMP were associated with improved GS versus the historical standard (SRR-SCS). No significant survival differences were observed between NRP and NMP, confirming that these strategies serve complementary roles in DCD LT. These findings support the adoption of both NRP and NMP as the new standard of care in DCD LT.

<h4>Objectives</h4>Ischemic stroke represents a major global cause of disability and death. Timely recanalization of occluded vessels to salvage the ischemic penumbra is the cornerstone of acute ischemic stroke (AIS) treatment. Nevertheless, the molecular mechanisms driving the transition from salvageable penumbra to infarct core in early AIS remain poorly understood. Recent evidence highlights the role of post-transcriptional methylation, especially N7-methylguanosine (m7G) modification, in stroke pathogenesis. This study aimed to systematically characterize transcriptome-wide m7G methylation changes in the ischemic penumbra during early AIS and to explore their potential functional relevance.<h4>Methods</h4>We performed middle cerebral artery occlusion in 8-week-old male BALB/c mice and applied MeRIP-seq to profile the transcriptome-wide m7G methylome in penumbra tissue. Bioinformatics analyses were conducted to elucidate the functional relevance of m7G-methylated transcripts.<h4>Results</h4>Compared with controls, AIS penumbra exhibited significant alterations in m7G-modified mRNAs. Specifically, upregulated m7G modifications were notably enriched in mRNAs associated with the mitogen-activated protein kinase (MAPK) signaling pathway, whereas downregulated modifications were significantly linked to axon guidance pathways.<h4>Conclusions</h4>This study provides the first systematic landscape of m7G methylation in AIS and m7G RNA methylation-related alterations in the MAPK signaling pathway may serve as potential therapeutic targets for stroke intervention.

Fasting enhances small intestinal regeneration after radiation, but the contribution of the gut microbiome to this process remains uncharacterized. We identify <i>Akkermansia muciniphila</i> (<i>AKK</i>) as a key mediator of this response. <i>AKK</i> was enriched in fasted mice and its antibiotic depletion abrogated radioprotection, whereas reintroduction restored both organismal survival and intestinal integrity. Fasting elevated propionic acid, consistent with <i>AKK</i>'s metabolic output. <i>AKK</i>-conditioned medium and propionate induced histone H3 acetylation in intestinal stem cell cultures while in vivo fasting induced <i>AKK</i>-dependent H3K27ac and H3K9ac, remodeling promoter-enhancer landscapes in crypt epithelial cells. Epigenetic profiling revealed a rewired core regulatory program enriched for pioneer transcription factors (Foxa, Gata, Klf), architectural organizers (Ctcf, Boris), and lineage-defining and metabolic regulators (Cdx2, Hnf4). This program supports expansion of a population of primed persister cells characterized by open chromatin accessibility at key stem and regenerative-associated loci including <i>Clu</i>, <i>Olfm4</i>, <i>Lgr5, Ascl2, Lrig1, Sox9, Rnf43, and Axin2.</i> These findings define a fasting-induced microbiome-metabolite-chromatin axis that epigenetically primes highly plastic persister cells for rapid regeneration of the intestinal epithelium following radiation-induced injury.

DNA offers exceptional information density and long-term stability, yet its practical deployment is limited by destructive readout and the absence of a reusable, physically addressable architecture that connects nanoscale molecular information with macroscale device-level data organization. Here, we present a regenerative Living Disk-Drive system based on thermo-responsive engineered living memory microspheroids (ELMMs), in which data-encoded bacteria are encapsulated as discrete, file-level living storage units. Each ELMM contains a clonal bacterial population carrying both an information plasmid, which encodes 26 × 26 pixel icon payloads and one- to three-color intracellular fluorescent retrieval indices, and a help plasmid that enables CRISPR-Cas12a/λ-Red rewriting of the data sequence and retrieval tag. A lyophilized ELMM database forms the Living Disk, which is coupled to an Optical Retriever and desktop-scale Living Drive for closed-loop retrieval, regeneration, and database replenishment. Released bacteria regrow for downstream readout or rewriting, while a fraction is re-encapsulated into new ELMMs. The tested system retains retrieval, regrowth, and sequence recovery after four months of ambient dry storage and 13 lyophilization-rehydration cycles. Model-based performance estimates are reported only as theoretical architecture-level bounds. These results establish an experimentally bounded yet extensible architecture for physically manageable and regenerative DNA memory.

<h4>Background and objectives</h4>Myelin oligodendrocyte glycoprotein antibody-associated disease (MOGAD) is an inflammatory demyelinating disorder that overlaps clinically with multiple sclerosis (MS) but immunopathologically distinct. Although often considered an acute inflammatory disease, recurrent attacks in MOGAD can lead to demyelination, axonal injury, and secondary neurodegeneration. Reliable biomarkers associated with relapse risk and disease subphenotypes, including optic neuritis, remain limited. Here, we aimed to define molecular and cellular signatures that distinguish MOGAD from MS as a prototypical neuroinflammatory disease and from Alzheimer disease (AD) as a proxy of neurodegeneration and to identify candidate immune-proteomic features associated with relapse frequency and clinical phenotype in MOGAD.<h4>Methods</h4>CSF, serum, and whole-blood samples from patients with MOGAD (n = 67), MS (n = 49), and AD (n = 36) were profiled using NULISAseq™ CSF proteomics, Olink Explore 3072 CSF and serum proteomics, and high-dimensional mass cytometry for immune cell characterization. In MOGAD, longitudinal clinical data, including total attack counts from the earliest documented attack through follow-up, were integrated with immune and proteomic profiles to assess associations with disease course and clinical phenotype.<h4>Results</h4>CSF and blood proteomic profiling revealed distinct inflammatory and cardiometabolic proteomic profiles in MOGAD, differentiating it from both MS and AD. Compared with MS, MOGAD showed relative reductions in lymphocyte populations with regulatory phenotypes. Within MOGAD, relapsing disease was associated with reduced frequencies of CD8⁺CCR7⁺CD31⁺CTLA4⁺ T cells and concurrent expansion of double-negative γδ T-cell subsets. IL-13 correlated positively with relapse frequency and inversely with circulating regulatory T cells, whereas IL-32 and CASP4 showed opposite associations, correlating negatively with relapse count and positively with Treg frequency. IL-13 was also inversely associated with CD31-expressing CD8⁺ T cells. Phenotype-stratified analyses suggested that these immune-proteomic relationships differed according to clinical presentation, including optic neuritis vs nonoptic neuritis phenotypes.<h4>Discussion</h4>This integrative immune-proteomic analysis identifies cellular and molecular features associated with relapsing vs monophasic MOGAD, suggesting a model of impaired peripheral immune regulation in relapsing disease. While exploratory, these findings generate a concrete hypothesis for future longitudinal and functional studies aimed at refining biomarker-based monitoring and informing individualized therapeutic strategies in MOGAD.

<h4>Background</h4>Parkinson's disease (PD) and cutaneous melanoma show epidemiological co-occurrence, but their shared genetic architecture remains unclear METHODS: We combined bidirectional time-dependent cox regression in the UK biobank (N ≈ 500,000) with a multi-layered genomic analysis using large-scale PD and melanoma GWAS summary statistics (N = 745,746). Genetic analyses included linkage disequilibrium score regression (LDSC), bidirectional Mendelian randomization (MR), PLACO, LAVA, and Bayesian colocalization RESULTS: Melanoma was associated with increased subsequent PD risk (HR = 1.62, 95% CI: 1.18-2.23), whereas the reverse PD-to-melanoma analysis was not statistically significant and was limited by low event counts. LDSC showed a modest positive genetic correlation (rg = 0.181, P = 0.04), while MR provided no evidence of bidirectional causality. PLACO identified 134 pleiotropic SNPs across 25 gene loci, including SOX6, the GDF5-GSS-EDEM2 cluster, and immune-related loci at 17q21.32. LAVA identified 17 locally correlated loci, with antagonistic effects predominating (11/17, 64.7%). Bayesian colocalization highlighted SOX6 as the strongest shared-signal locus (PP·H4 = 0.904; PP·H3 = 0.054) CONCLUSIONS: PD-melanoma co-occurrence is supported by shared but heterogeneous genetic architecture rather than bidirectional causality. Antagonistic local effects may partially offset genome-wide correlation, while SOX6 represents a prioritized colocalized candidate locus linking dopaminergic neuron vulnerability and melanocytic biology.

γ-Secretase is a pivotal membrane-embedded protease that cleaves more than 150 single-span membrane proteins within their transmembrane domains. While γ-secretase is involved in a wide range of physiological processes, it is best known for its critical role in Alzheimer's disease, where it cleaves a C-terminal fragment of the amyloid precursor protein into small aggregation-prone and neurotoxic peptides. However, how γ-secretase recognizes and recruits its substrates, how it binds and unfolds them, where drug-binding sites are located, and what the full range of its substrates and functions is, have all remained unknown. These long-standing questions have been at the forefront of research for the past decade and are now increasingly being solved. In this review, we outline how recent advances in structural biology, biochemistry, and computational biology have helped to elucidate these mysteries. We also highlight future research directions needed to achieve a comprehensive understanding of this fascinating enzyme, a major therapeutic target for which Alzheimer's disease drugs are now on the horizon.

Lymphovascular space invasion (LVSI) is a critical factor linked to metastasis and poor outcomes in endometrioid endometrial carcinoma (EEC), yet its multicellular mechanisms remain unclear. Using single-cell RNA sequencing of 3 LVSI-present (LVSI+) EECs, 2 LVSI-absent (LVSI-) EECs, and 2 normal endometrial samples, we delineated the cellular ecosystems underlying LVSI. LVSI+ EECs exhibited marked epithelial reprogramming, transitioning from differentiated ciliated epithelium to hyperproliferative and metabolically remodelled phenotypes, and contained TC4, a metastatic epithelial subset characterized by hypoxia, partial epithelial-mesenchymal transition, immunosuppression, and progesterone resistance. We also identified nine key genes in malignant epithelial cells associated with LVSI prognosis. The tumour microenvironment in LVSI+ EECs shifted from an inflammatory state dominated by epithelial cells to a collaborative network involving stromal and immune cells. This network was enriched in immunosuppressive ZNF683 + SOX4 + CD8+ T cells, Cycling_T cells, SPP1 + MMP9 + Mac, WNT5A_mCAF, Tip endothelial cells, and lymphatic endothelial cells. WNT5A_mCAF and SPP1 + MMP9 + Mac synergistically remodelled the extracellular matrix, promoted lymphangiogenesis and angiogenesis, and, together with endothelial cells, suppressed T-cell activity via LGALS9-HAVCR2/CD44 inhibitory signalling, thereby establishing a niche conducive to vascular invasion. Using spatial multiplex immunofluorescence, we confirmed hypoxic tumour epithelial cells at the invasive front coexisting with an immunosuppressive microenvironment, and revealed spatial colocalization of PD-L1+ tumour cells, PD-L1+ macrophages, and PD-1+ T cells within LVSI thrombi. Our comprehensive study provides deeper insights into LVSI as an actively coordinated multicellular process, potentially improving LVSI risk prediction, supporting treatment decision-making, and informing new therapies targeting the tumour microenvironment.

<h4>Aims/hypothesis</h4>G-protein-coupled receptors (GPCRs) play an important role in maintaining systemic glucose homeostasis by regulating insulin secretion, with protein kinase A (PKA) signalling serving as a key downstream effector. Our previous work identified specific expression of type IIB PKA in pancreatic beta cells. Based on these findings, we propose that type IIB PKA is involved in mediating the GPCR signalling in pancreatic beta cells.<h4>Methods</h4>The glucagon-like peptide-1 (GLP-1) analogue liraglutide was administered to mice 30 min before glucose injection during an IPGTT, whereas the glucose levels and insulin levels were measured in wild-type and RIIβ-knockout mice. The isolated islets were subjected to both perifusion assay and static batch incubations following stimulation with liraglutide, glucagon and follicle-stimulating hormone (FSH). RNA-seq analysis was performed to identify molecular changes in islets with RIIβ ablation. Both western blotting and quantitative PCR were employed to quantify the gene expression. Whole-cell patch-clamp recordings were conducted to measure K_ATP and Ca²⁺ currents. Insulin granule morphology and abundance were evaluated by electron microscopy and flow cytometry using EGFP-labelled Syncollin, respectively.<h4>Results</h4>RIIβ-knockout mice exhibited impaired glucose tolerance and attenuated insulin secretion in response to liraglutide. Islets isolated from RIIβ-knockout mice showed reduced insulin secretion following liraglutide stimulation. Similarly, RIIβ-ablated islets displayed decreased insulin secretion in response to both glucagon and FSH. Further mechanistic studies revealed that RIIβ deficiency impaired liraglutide-mediated PKA signalling activation. Specifically, RIIβ-ablated beta cells exhibited reduced basal K_ATP channel activity and lack of liraglutide-mediated channel inhibition. Multiple voltage-gated Ca²⁺ channel genes were downregulated in RIIβ-ablated islets, leading to a mild reduction in basal Ca²⁺ current and a significant decrease following liraglutide treatment. RIIβ-knockout beta cells also exhibited reduced insulin granule size, decreased total granule number and fewer granules docked at the plasma membrane.<h4>Conclusions/interpretation</h4>Our results highlight type IIB PKA as a primary mediator of Gs-coupled receptor-potentiated insulin secretion, providing a new molecular framework for metabolic regulation research.

Alzheimer's disease (AD) is characterized by progressive cognitive decline accompanied by synaptic dysfunction and neuronal loss. Dysregulation of specific microRNAs (miRNAs) has been increasingly implicated in AD pathogenesis, suggesting that miRNA-mediated regulatory pathways may represent important mechanisms underlying neuronal vulnerability. Here, we identified mmu-microRNA-204-5p (miR-204) as a critical regulator of neuronal survival that is markedly downregulated in the hippocampus of 5xFAD mice. Through integrated bioinformatics analysis and experimental validation, we established neuronal pentraxin-1 (NPTX1) as a direct post-transcriptional target of miR-204. In primary cultured neurons, exposure to Aβ₁₋₄₂ oligomer (oAβ) drove marked upregulation of NPTX1 and precipitated neuronal injury, manifested by aberrant reactive oxygen species (ROS) accumulation, collapse of mitochondrial membrane potential, diminished cell viability, and dendritic degeneration. Inhibition of miR-204 further exacerbated these pathological changes and engaged mitochondrial apoptotic signaling, as evidenced by Bax upregulation, cytochrome c release, and caspase-3 cleavage. Conversely, restoration of miR-204 expression or genetic knockdown of NPTX1 attenuated oxidative stress, preserved mitochondrial integrity, and restored neuronal survival. Collectively, our findings uncover a previously unrecognized miR-204-NPTX1 regulatory axis that governs mitochondrial integrity and apoptotic susceptibility in AD, highlighting miR-204 as a potential therapeutic target for AD and related oxidative stress-associated neurodegenerative disorders.

Tumor cells face constant environmental and therapeutic stresses, relying on interconnected survival mechanisms involving autophagy, metabolism, and cell death regulation. This review explores the triangular relationship between these processes and how their dynamic interplay supports tumor growth and drives resistance to various treatments. We discuss key molecular pathways regulating autophagy and metabolic reprogramming, and their impact on different cell death modalities. Finally, we address current challenges in translating this knowledge into clinical therapies, emphasizing the need for precise biomarkers and combination strategies to overcome resistance. Understanding this complex network offers promising avenues for developing more effective cancer treatments.

In the title mol-ecule, C₃₄H₃₇F₅O₅, the dihedral angles between the central carbonyl-phenyl and adjacent perfluoro-phen-oxy and (tetra-dec-yloxy)benzoate rings are 74.19 (2) and 67.86 (2)°, respectively and the tetra-decyl chain adopts an extended conformation. In the crystal, the mol-ecules are linked by C-H⋯O and C-H⋯F hydrogen bonds, forming <i>C</i>(7) and <i>C</i>(10) chains, respectively, both running infinitely along [010]. The Hirshfeld surface analysis reveals that the major contributions to the two dimensional fingerprint plots are from H⋯H (49.4%), F⋯H/H⋯F (16.7%) and O⋯H/H⋯O (9.0%) contacts. An inter-molecular inter-action energy calculation shows that dispersion energy contributes the most to the consolidation of the structure.

<h4>Background</h4>Intrahepatic Cholangiocarcinoma (ICC) is a highly malignant tumor originating from the intrahepatic biliary epithelium, characterized by strong invasiveness and poor prognosis. For patients with unresectable advanced ICC, the current first-line standard treatment is the gemcitabine plus cisplatin regimen, but its efficacy is limited. In recent years, with the exploration of immunotherapy, targeted therapy, and various combination treatment modalities, new directions have been provided for the treatment of advanced ICC.<h4>Case presentation</h4>This article reports a case of a patient with initially unresectable ICC who underwent conversion therapy with percutaneous portal vein embolization (PVE) combined with surufatinib (a multi-kinase inhibitor targeting VEGFR, FGFR, and CSF-1R), sintilimab (a PD-1 inhibitor), and the FOLFOX regimen (leucovorin, fluorouracil, and oxaliplatin). This approach successfully enabled radical liver resection and resulted in long-term disease-free survival. The patient was a 71-year-old male diagnosed via enhanced CT and MRI with a large liver mass accompanied by portal vein tumor thrombus. Pathological confirmation by needle biopsy confirmed intrahepatic cholangiocarcinoma. Due to extensive tumor invasion and insufficient future liver remnant (FLR), the tumor was assessed as unresectable. The treatment team adopted a multimodal conversion strategy: first performing PVE to promote compensatory hyperplasia of the FLR; subsequently administering 4 cycles of surufatinib, sintilimab combined with FOLFOX chemotherapy. Post-treatment imaging evaluation showed significant tumor shrinkage, along with an increase in FLR volume, meeting the criteria for safe radical surgery. Two weeks after stopping medication, the patient underwent right hemihepatectomy, caudate lobe resection, cholecystectomy, hepatoduodenal lymph node dissection and inferior vena cava repair. The patient recovered well postoperatively. At the last follow-up, he had achieved 28 months of disease-free survival with no evidence of local recurrence or distant metastasis.<h4>Conclusion</h4>For initially unresectable ICC, conversion therapy with PVE combined with surufatinib, sintilimab, and the FOLFOX regimen can significantly increase the tumor resection rate and achieve long-term survival. This strategy combines high anti-tumor efficacy with clinical feasibility, providing a new approach for the conversion therapy of advanced ICC. Further larger sample studies are needed in the future to verify its efficacy and safety.

The global rise in chronic inflammatory and autoimmune disorders has intensified research to understand cellular stress response pathways that drive immune dysregulation. Mitochondria have emerged not only as central hubs of cellular metabolism but also as active modulators of immunity and inflammation. Mitochondrial proteases are essential regulators of mitochondrial protein quality control, dynamics, and stress responses. By selectively degrading misfolded or damaged proteins, they maintain mitochondrial function and bioenergetic capacity. Beyond housekeeping roles, mitochondrial proteases also influence immune signaling by modulating mitochondrial stress pathways, reactive oxygen species production, and the release of mitochondrial-derived danger signals. Dysregulation of these proteases has been linked to chronic inflammation and contributes to the pathogenesis of inflammatory diseases. This review summarizes current knowledge on the role of mitochondrial proteases CLPXP, LONP1, i-AAA, m-AAA, as well as processing peptidase OMA1, in immune cells and inflammatory pathologies. We explore the molecular mechanisms by which these mitochondrial proteases regulate immune signaling, integrating the results from immune cells as well as other non-immune cell types, including those involved in cancer, neurodegeneration, renal injury, and other inflammatory pathologies. We explore mitochondrial proteases function as context-dependent regulators of immunometabolic signaling, with effects shaped by cell type, metabolic state, and stress conditions. Finally, we discuss emerging small molecules and drugs targeting mitochondrial proteases to highlight their potential therapeutic role in modulating inflammation. By situating mitochondrial proteases at the crossroads of immunometabolism and therapeutic intervention, this review underscores their untapped potential in the development of innovative anti-inflammatory strategies.

Adipose tissue remodeling is a dynamic process essential for metabolic homeostasis, enabling tissue expansion, extracellular matrix (ECM) turnover, angiogenesis, and coordinated immune adaptation. In obesity, however, maladaptive remodeling characterized by fibrosis, chronic low-grade inflammation, and hypoxia disrupts adipose plasticity and promotes systemic insulin resistance. Central to these processes is the tightly regulated homeostasis between proteases and their inhibitors, in which the serine protease inhibitor (serpin) superfamily represents an important yet underappreciated regulatory axis. Beyond their classical roles in coagulation and fibrinolysis, serpins regulate ECM remodeling, macrophage recruitment and polarization, cytokine signaling, angiogenic responses, adipokine activity, and insulin sensitivity, thereby orchestrating immune-metabolic crosstalk within adipose depots. Emerging evidence indicates that individual serpins exert distinct and context-dependent effects, with some promoting fibrosis, inflammation, and metabolic dysfunction, whereas others preserve adipose tissue homeostasis and metabolic function. This review synthesizes current knowledge on the structural and functional diversity of the serpin superfamily and examines their mechanistic roles in adipose tissue remodeling during obesity, with particular emphasis on how adipose-associated serpins regulate adipose tissue homeostasis, depot-specific remodeling, and immune-metabolic crosstalk. The review further discusses the experimental and translational applications of emerging single-cell and spatial transcriptomics, multi-omics, and computational approaches that may advance the understanding of serpin biology, improve the investigation of human adipose tissue, and accelerate the identification of clinically relevant serpin-related biomarkers and therapeutic targets for obesity and related metabolic disorders. By positioning serpins as key regulators of adipose tissue remodeling and immune-metabolic integration, this review highlights protease-antiprotease balance as a central determinant of metabolic health and identifies serpins as promising biomarkers and therapeutic targets for obesity and related metabolic disorders.

The uplift of the Qinghai-Tibet Plateau has shaped a unique extreme environment, fostering distinct endemic aquatic organisms. <i>Schizopygopsis pylzovi</i>, a vulnerable endemic fish in the upper Yellow River, is a key model for studying the biogeographic patterns of plateau fish. To assess its genetic characteristics and evolutionary dynamics, we comprehensively evaluated 11 geographic populations of <i>S. pylzovi</i> using two complementary mitochondrial markers, the conserved COI gene and the fast-evolving D-loop region. A total of 142 COI and 143 D-loop sequences were analyzed, and sequences alignment, haplotype network construction, AMOVA, and neutrality tests were performed. AMOVA revealed that genetic variation was mainly distributed within populations, indicating weak population differentiation. Neutrality tests and mismatch distribution analysis suggested historical and recent population expansion events. Our findings highlight the value of joint analysis using COI and D-loop markers in revealing the genetic structure of <i>S. pylzovi</i>, provide new insights into the impact of plateau uplift on fish evolution, and establish a scientific basis for the conservation of this vulnerable species.

Endothelial dysfunction is a central pathophysiological mechanism in the development of cardiovascular disease (CVD), the leading global cause of morbidity and mortality. Vascular homeostasis critically depends on the endothelial glycocalyx, which regulates vascular permeability, mechanotransduction, and anti-inflammatory signaling. Heparanase, the only known mammalian endoglycosidase capable of cleaving heparan sulfate, plays a pivotal role in glycocalyx degradation, a process exacerbated by oxidative stress, hyperglycemia, and hypertension. Excessive heparanase activity has been implicated in several cardiovascular pathologies, including heart failure, ischemia-reperfusion injury, and atherosclerosis, yet effective therapeutic strategies remain limited. Emerging evidence suggests that vitamin D may preserve endothelial integrity by maintaining the structure of the glycocalyx and suppressing heparanase expression, as demonstrated in podocytes. By attenuating heparanase-mediated degradation of the glycocalyx in the vasculature and heart, vitamin D could exert protective effects against endothelial dysfunction and thereby mitigate cardiovascular risk.

Prostate cancer (PCa) is among the highest incidence malignancies in men, with high rates of inevitable resistance development, relapse, and mortality. Castration-resistant prostate cancer (CRPC) continued to pose substantial therapeutic challenges, highlighting the urgent need for effective treatment options. This study assessed the marine cembranoid sarcophine activity against the progression and recurrence of the metastatic CRPC (mCRPC) in mouse xenograft models. Protein and phosphorylation levels were assessed by immunoblotting and mRNA expression by qPCR and RNA sequencing. The in vivo efficacy was evaluated through tumor progression over 3 weeks followed by primary tumor excision and recurrence monitoring over an 8-week course. Sarcophine significantly reduced the mCRPC CWR-R1ca tumor volume by 74.1% and suppressed the epigenetic regulators EZH2 and SMYD2; lineage plasticity factors ASCL1 and BRN2; Wnt/stemness signaling markers β-catenin and LGR6; AKT total expression and activation; and invasion-associated proteins TRPC4 and MMP2 in primary tumors. Sarcophine effectively prevented the mCRPC locoregional recurrence, as well as lung and spleen distant recurrences, and effectively reduced recurrence in other organs. Transcriptomics-RNA-Seq analysis of primary tumors identified 2697 downregulated and 3534 upregulated genes, indicating broad transcriptional reprogramming following sarcophine treatments. These findings demonstrate coordinated suppression of multi-oncogenic pathways and validate the therapeutic potential of sarcophine to control mCRPC.

Osteoarthritis (OA) and osteoporosis (OP) are prevalent conditions with a complex relationship, yet their shared epigenetic mechanisms remain poorly understood. While genes like <i>DKK1</i>, <i>RHOJ</i>, and <i>SOX6</i> have been implicated in both diseases, the specific role of individual CpG sites has not been fully characterized. We investigated CpG methylation in these genes using bisulfite pyrosequencing of peripheral blood DNA from n = 96 postmenopausal women: n = 24 with comorbid OA and OP, n = 34 with OA, and n = 38 healthy controls. Methylation differences were analyzed using statistical tests and logistic regression. Comorbid patients showed significant hypermethylation at two <i>DKK1</i> CpG sites compared to the OA-only group (p_adj = 0.0007 and p_adj = 0.042). Conversely, one <i>DKK1</i> site was hypomethylated in the OA-only group relative to controls (p_adj = 0.03). A regression model combining three <i>DKK1</i> sites and one <i>SOX6</i> site demonstrated predictive value for comorbid disease, with an AUC of 0.696. These findings identify site-specific methylation of <i>DKK1</i> and <i>SOX6</i> as a molecular signature associated with comorbid OA and OP, offering new insights into their shared etiology.

Voltage-gated ion channels (VGICs) are traditionally associated with electrically excitable cells; however, increasing evidence indicates that they are also expressed in non-excitable cells, including vascular endothelial cells. This review aims to summarize the current knowledge on the expression, regulation, and functional role of VGICs in the vascular endothelium, and to highlight their potential contribution to endothelial signaling. We examined the molecular structure, biophysical properties, and functional roles of voltage-gated Na⁺ (Na_V), Ca²⁺ (Ca_V), and K⁺ (K_V) channels in vascular endothelial cells. Particular attention was given to studies investigating VGIC activity in native endothelium and to emerging mechanisms regulating their activation. Endothelial cells express multiple VGIC subtypes at low densities, which are insufficient to generate action potentials but can modulate membrane potential (V_M) and Ca²⁺-dependent signaling. The dynamic regulation of the endothelial V_M, through the interplay between hyperpolarizing and depolarizing conductances, emerges as a key determinant of VGIC availability and activation. VGICs contribute to essential endothelial functions, including angiogenesis, vasomotor responses, blood-brain barrier permeability, and inflammation. Dysregulated VGIC expression and/or activity may be implicated in several pathological conditions, such as atherosclerosis, calcific aortic stenosis, and tumor vascularization. VGICs represent an unexpected but functionally relevant component of endothelial signaling. Elucidating their role in native vascular beds and disease contexts may uncover novel mechanisms of endothelial regulation and identify new therapeutic targets in cardiovascular and cancer biology.

Across all kingdoms of life, ribosomes are indispensable molecular machines that translate genetic information into the proteome of living cells. The fundamental catalytic centers of the ribosome, constructed primarily from ribosomal RNA (rRNA), exhibit remarkable conservation between the major domains of life. The ribosome's A-site deciphers the mRNA's triplet code, while the P-site synthesizes the growing protein chain and the E-site provides exit for deacylated tRNA; a distinct tunnel facilitates nascent polypeptide export. While the conservation of ribosomal proteins is less pronounced between bacteria and eukaryotes, striking homology exists from simple eukaryotes to humans. Ribosomal proteins were traditionally viewed mainly as scaffolding agents, steering rRNA folding during ribosome biogenesis and maintaining structural stability during translation. However, since the early 2000s, advances in structural and functional ribosome analysis have ushered in a more nuanced paradigm: ribosomes are no longer considered uniform machines. Instead, an array of rRNA and ribosomal protein modifications generates a spectrum of ribosome populations capable of specialized translation. RiboScreen^TM technology leverages this regulatory potential of individual ribosomal proteins, enabling deliberate modulation of target protein output and representing a promising tool for correcting dysregulated protein expression involved in rare and common diseases. This review will first introduce relevant aspects of ribosome biology and then showcase the tools of this new technology. Finally, we report examples for the delivery of small molecules to target ribosomal proteins for tailored restoration of protein production levels in rare and prevalent diseases.

<title>Abstract</title> <p> Colorectal cancer (CRC) tumours with high stromal content have a worse outcome, but the mechanisms governing this are unclear. Using high-throughput single-cell perturbation analysis of CRC patient-derived organoids (PDOs) and cancer-associated fibroblasts (CAFs) we find that epithelial cells with high stromal-communication potential are marked by the transcriptional co-repressor <italic>DACH1</italic> . To define the causal regulators of stromal-epithelial signalling, we developed a novel CRISPR screening platform to perturb the CAF secretome and measure epithelial stem cell responses at single-cell resolution. Intercellular CRISPR screening and full factorial ligand analysis revealed that stromal Prostaglandin E2 (PGE ₂ ) is a dominant regulator of CRC cell-fate plasticity. Stromal PGE ₂ converts <italic>DACH1</italic> ⁺ epithelia from a chemosensitive proliferative colonic stem cell (proCSC) fate into a chemorefractory and pro-metastatic revival colonic stem cell (revCSC) fate. PGE ₂ -driven epithelial transdetermination is rapid and reversible, providing an acute mechanism for stromal-driven plasticity in CRC tumours. Genetic and pharmacological inhibition of stromal COX2 inhibits epithelial plasticity, trapping CRC epithelia in an anti-metastatic and chemosensitive proCSC fate. <italic>PTGS2</italic> ⁺ CAFs support a spatially resolved revCSC to proCSC plasticity gradient in human CRC tumours marked by increasing <italic>DACH1</italic> expression. These results reveal that stromal prostaglandin is a dominant spatial regulator of poor-prognosis cell-fates and may explain the benefit of anti-COX therapies in both preventing and treating CRC. </p>

Excessive fructose consumption is increasingly linked to obesity, diabetes, non-alcoholic fatty liver disease (NAFLD), and cardiovascular disease, yet its impact on brown adipocytes remains poorly defined. Here, we investigated how fructose alters brown adipogenesis with a focus on thyroid hormone receptor (THR) signaling. Differentiation of mouse brown preadipocytes in fructose-containing medium resulted in markedly reduced lipid droplet size and number, impaired expression of adipogenic markers (PPARγ, FABP4), and a loss of the brown adipocyte identity protein UCP-1. Whereas triiodothyronine (T3) and the THRβ-selective agonist resmetirom dose-dependently enhanced UCP-1 expression in glucose medium, neither T3 nor resmetirom restored UCP-1 in fructose medium, revealing thyroid hormone resistance. Mechanistically, fructose specifically downregulated THR but not its heterodimer partner, retinoid X receptor (RXR), via ubiquitin-mediated degradation. Overexpression of THR rescued UCP-1 in glucose medium and promoted dynamic lipid turnover but failed to restore UCP-1 or lipid remodeling in fructose medium, indicating a defect in THR transcriptional regulation. Our secretome proteomics revealed that both the sugar source and THR overexpression strongly remodel the secreted protein landscape of brown adipocytes. The data indicate that fructose both reconfigures metabolic and signaling networks and blunts oxidative resilience, while excess THR activity in fructose conditions disrupts energy metabolism and increases the risk of protein misfolding. All these could contribute to THR dysfunction and impair adipogenesis. Collectively, these findings identify fructose as a potent inhibitor of brown adipogenesis, suppressing THR-dependent UCP-1 expression by inducing thyroid hormone resistance through a defect in THR-mediated transcriptional regulation. This mechanism links dietary fructose to impaired brown adipocytes and metabolic imbalance.

Axonal growth cones rely on local proteomic changes to interpret guidance cues while navigating towards their synaptic targets. Yet the developmental dynamics of their proteome remain poorly understood. By isolating growth cones from the whole mouse hippocampus and dentate gyrus at early postnatal days, we performed a systematic characterization of their proteome. We show that growth cones from both subregions share a proteomic signature at postnatal day (P) 1, dominated by energy metabolism and protein synthesis, before taking distinct developmental trajectories. In fact, hippocampal growth cones undergo a rapid transition, from RNA-related processes to synapse formation. Dentate gyrus growth cones, however, maintain a similar proteomic profile at P3 and P5, suggesting a prolonged exploratory behaviour, consistent with later ingrowth of entorhinal cortex fibres. At these early timepoints, hippocampal and dentate gyrus growth cones are characterized by several temporally regulated proteins, suggesting dynamic and different developmental trajectories. Our dataset provides a comprehensive proteomic resource for understanding growth cone function during hippocampal circuit formation and insights into the molecular mechanisms underlying region-specific developmental timelines.

Mitochondria-associated endoplasmic reticulum membrane (MAM), which serves as a signaling hub for interactions between the endoplasmic reticulum (ER) and mitochondria, dynamically coordinates innate immune processes by regulating calcium homeostasis, lipid metabolism, mitochondrial dynamics, mitochondrial protein modifications, and autophagy. MAM regulates calcium homeostasis to govern mitochondrial energy metabolism and inflammasome activation; maintains lipid metabolism for membrane integrity to support antiviral signaling pathways; controls mitochondrial fission and fusion dynamics, processes that are closely associated with mitochondrial DNA (mtDNA) release; regulates mitochondrial protein modifications to fine-tune the function of proteins localized at MAM; and facilitates the clearance of damaged mitochondria and leaked mtDNA through autophagy. Most critically, MAM dysfunction and innate immune dysregulation form a vicious cycle: immune activation disrupts MAM integrity, and MAM abnormalities exacerbate the release of mitochondrial damage-associated molecules, continuously driving overactivation of pathways such as inflammasomes and the cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) pathway, thereby promoting the development of autoimmune diseases. This review synthesizes current literature on the molecular mechanisms by which MAM regulates innate immunity. We summarize how disruptions in MAM-mediated mitochondrial homeostasis contribute to innate immune imbalance. By integrating these findings, we highlight potential intervention nodes. This underscores the clinical relevance of targeting MAM in immune-related pathological conditions.

<h4>Background</h4>Clear cell renal cell carcinoma (ccRCC) is the most common subtype of renal malignancy and remains a major cause of cancer-related mortality worldwide. Although advances in surgery, targeted therapy, and immunotherapy have improved outcomes for patients, reliable biomarkers for predicting prognosis remain limited. Therefore, robust gene-based prognostic models are urgently needed to improve risk stratification and guide individualized treatment strategies.<h4>Methods</h4>We developed a novel prognostic model integrating apoptosis and immune - related genes (AIRGs) to predict overall survival (OS) in patients with ccRCC.<h4>Result</h4>Using Gene Set Enrichment Analysis (GSEA) combined with least absolute shrinkage and selection operator (LASSO) Cox regression, we identified 7 key prognostic genes, namely, CCR4, TNFRSF10A, TEK, TGFA, CD14, IFITM1, and SEMA3G, that collectively demonstrated strong predictive performance in TCGA cohort with c-index = 0.711. Functional enrichment analyses revealed that apoptosis, immune regulation, and multiple oncogenic signaling pathways were significantly associated with the risk score, highlighting the critical role of the tumor microenvironment in ccRCC progression. Transcription factor binding analysis based on the JASPAR database suggested that RELA and STAT3 with scores of 0.829 and 0.951, respectively are potential upstream regulators within the prognostic network, particularly influencing TNFRSF10A expression. External validation using the International Cancer Genome Consortium (ICGC) dataset confirmed the robustness of the prognostic model with c-index = 0.612 Furthermore, in vitro experiments demonstrated that RELA and STAT3 regulate TNFRSF10A-mediated apoptotic signaling in ccRCC cells, providing mechanistic support for the bioinformatic findings.<h4>Conclusion</h4>This study establishes a biologically informed and clinically relevant prognostic framework for ccRCC. Our findings highlight the therapeutic potential of targeting the RELA/STAT3-TNFRSF10A axis and contribute to the advancement of precision medicine in ccRCC.

<h4>Background</h4>Sarcoidosis is a systemic granulomatous disease with heterogeneous clinical manifestations and unclear pathogenesis. Although genome-wide association studies (GWAS) have identified several immune-related loci, the functional interpretation of these signals remains limited. Integrative transcriptome-wide approaches may uncover novel susceptibility genes and provide mechanistic insights.<h4>Objectives</h4>The primary objective of this study was to identify novel susceptibility genes for sarcoidosis and provide mechanistic insights into its pathogenesis through a comprehensive, cross-tissue transcriptome-wide approach.<h4>Methods</h4>We conducted a cross-tissue transcriptome-wide association study (TWAS) using the UTMOST framework, followed by single-tissue TWAS via FUSION. Candidate genes identified in both analyses were further evaluated using Multi-marker Analysis of Genomic Annotation (MAGMA), Mendelian randomization (MR), and Bayesian colocalization. Functional characterization was explored through gene-chemical-disease associations from the Comparative Toxicogenomics Database (CTD) and phenome-wide association studies (PheWAS) in the UK Biobank.<h4>Results</h4>Cross-tissue TWAS identified 48 genes. Integrative analyses prioritized four novel susceptibility genes: RNF215, PLCL1, FAM117B, and RFTN2. MR and colocalization supported causal effects of RNF215 (risk-increasing), FAM117B and RFTN2 (protective), and tissue-dependent effects for PLCL1. CTD analyses revealed interactions of these genes with environmental chemicals including bisphenol A and tetrachlorodibenzodioxin, while PheWAS demonstrated pleiotropic associations with immune, respiratory, hematological, and cardiovascular traits.<h4>Conclusions</h4>This comprehensive integrative study identifies four biologically plausible susceptibility genes for sarcoidosis, expanding the genetic architecture of sarcoidosis and suggest potential targets for mechanistic and therapeutic investigation.

Serotonin (5-hydroxytryptamine; 5-HT) is a key neuromodulator involved in the regulation of mood, appetite, aggression, and impulse control. Dysregulation of central 5-HT function has been implicated in alcohol use disorder (AUD) and comorbid depression. This review summarizes clinical and preclinical evidence on the role of 5-HT in AUD development, heterogeneity, and treatment response. Alterations in 5-HT function may be shaped by stress and genetic variation in serotonergic genes, including TPH2 and SLC6A4, contributing to individual vulnerability to AUD. Reduced 5-HT activity increases the risk of developing AUD, particularly Cloninger's type II, characterized by early onset, violent, and antisocial behaviors. Consistently, Tph2-deficient mice, which lack central 5-HT, exhibit increased ethanol consumption and behavioral features resembling Cloninger's type II alcohol dependence. Similarly, alcohol-preferring rat lines show reduced 5-HT levels, decreased serotonergic projections to the cortex, and reduced prefrontal 5-HT_2A receptor binding, implicating raphe-prefrontal serotonergic projections as a critical pathway modulating vulnerability to AUD. Given the heterogeneity of AUD, the efficacy of selective 5-HT reuptake inhibitors (SSRIs) remains limited, with beneficial effects observed only in less severe, later-onset forms. Recently, serotonergic psychedelic-assisted therapies have attracted considerable interest as potential AUD treatments; mechanistically, their effects may be linked to activation of 5-HT_2A receptors in the prefrontal cortex - a brain region known to be dysfunctional in AUD. The reviewed literature highlights the need for improved stratification of AUD subtypes and validation of 5-HT-related biomarkers to guide personalized therapeutic approaches.

Cell-type deconvolution has been instrumental for the analysis of spatial transcriptomics (ST) data to reveal underlying tissue heterogeneity. Although reference-based methods have been widely explored, practical limitations, particularly the need for matched single-cell RNA-seq data sets, highlight the value of robust reference-free methods. Existing reference-free approaches, such as STdeconvolve, overlook spatial information, despite the well-established observation that spatially adjacent spots often share similar cellular compositions. Motivated by this, we propose SpatialCD, a spatially informed reference-free deconvolution method that extends Latent Dirichlet Allocation (LDA) with spatial regularization to encourage neighboring spots to exhibit similar cell-type structures. SpatialCD produces improved estimates of cell-type proportions and gene expression profiles. Across simulated and real data sets, including MERFISH-derived simulations, mouse olfactory bulb (MOB), 10× Visium, and DBiT-seq data, SpatialCD consistently improves performance over existing reference-free methods across evaluated data sets by recovering more accurate transcriptional patterns and revealing biologically coherent spatial organization across normal and diseased tissues, including subtle anatomical layers and region-specific tumor-associated cell populations. This work advances statistical tools for spatial transcriptomics and enriches the methodological toolkit for complex spatial gene expression analysis.

Cells release heterogeneous extracellular vesicles and particles (EVPs) into circulation, carrying RNA and proteins that reflect their origin. Recently, brain-derived EVs have gained significant attention as non-invasive biomarkers for Alzheimer's disease (AD). Here, we identified sub-50nm extracellular nanoparticles in human brain and blood that lack the hallmarks of small EVs, exosomes, exomeres, and supermeres but are enriched for brain-specific markers, hereafter termed small EPs or 'SECmeres'. We discovered that RNAs associated with SECmeres discriminated AD cases from controls with higher significance than small EVs, large EVs showed no differences. Discriminating RNAs were enriched in small EVs (Synaptotagmin, Alpha-synuclein, MAPT) or SECmeres (L1CAM, Syntaxin, Neurogranin), indicating distinct brain-derived signatures. Single-cell RNAseq deconvolution shows small EVs contain RNAs from diverse brain cells, whereas SECmeres enrich brain endothelial transcripts, lining cerebral blood vessels and forming the blood-brain barrier (BBB). These findings challenge the prevailing view that small EVs are the primary carriers of biomarkers. Collectively, our study shows that blood EVPs carry brain-specific information for liquid biopsy, pending validation in larger blinded clinical trials.

Brown adipose tissue (BAT) counteracts obesity-related metabolic dysfunction through both thermogenic and non-thermogenic means. However, substantial evidence indicates that obesity negatively affects BAT mitochondrial morphology and oxidative capacity, impairing systemic energy homeostasis. Motivated by this apparent contradiction, we investigate the relationship between obesity and mitochondrial dynamics, as the underlying mechanisms remain incompletely understood. Here, we identify E4BP4 as a transcriptional repressor that prevents obesity-induced mitochondrial fragmentation and oxidative dysfunction by inhibiting ceramide synthesis in brown fat. Specifically, E4BP4 interacts with PRDM16 to repress Cers6 mRNA expression and consequently reduces C16:0 ceramide levels by binding to a 65 kb upstream enhancer region of the Cers6 gene. Notably, the preservation of mitochondrial integrity in BAT by E4BP4 gain-of-function improves systemic glucose homeostasis, independent of weight loss. Collectively, our findings establish E4BP4 as a molecular safeguard against obesity-induced mitochondrial fragmentation and oxidative dysfunction, primarily by suppressing ceramide synthesis in brown fat.

Atherosclerotic plaque rupture is a major cause of cerebrovascular events, yet the molecular determinants underlying vulnerability-related plaque morphology, including fibrous-cap thickness, remain incompletely defined. Using histomorphology-guided spatial proteomics, here we delineate molecular programs associated with plaque cap phenotype across discrete plaque subregions. In 112 carotid endarterectomy specimens, differences between thin-cap and thick-cap plaques were predominantly localized to the necrotic core and fibrous cap. These differences were enriched for processes related to inflammation, lipid handling, extracellular matrix remodeling and ossification/calcification, and supported the presence of proteome-based plaque subtypes. PCSK9 was among the proteins most strongly associated with thin-cap plaques. Consistently, an in vitro model of necrotic core-like oxidative and inflammatory stress increased PCSK9 secretion in primary vascular smooth muscle cells. Together, these findings localize molecular programs associated with cap phenotype to plaque compartments and provide a framework for spatially informed biomarker discovery in advanced carotid atherosclerosis.

<h4>Introduction</h4>Colorectal cancer (CRC) is one of the most prevalent malignancies worldwide, with up to 50% of patients developing metastatic disease, predominantly involving the liver. Hepatectomy remains the gold standard for resectable hepatic metastases; however, recurrence rates remain high and reliable prognostic biomarkers are lacking. Phosphorylated ribosomal protein S6 (pS6), a surrogate marker of PI3K/AKT/mTORC1 pathway activation, has been implicated in tumor aggressiveness across multiple cancer types, but its prognostic role after hepatectomy for metastatic colorectal cancer (mCRC) has not been established.<h4>Objective</h4>To evaluate the prognostic impact of pS6 expression and other molecular markers (BRAF V600E, p53, CERBB2, MSH6, PMS2, FUS, and HSPA9) assessed by immunohistochemistry on tissue microarray (TMA) on the clinical outcomes of patients undergoing hepatectomy for metastatic colorectal cancer.<h4>Materials and methods</h4>A retrospective cohort study was conducted including 64 patients who underwent hepatectomy for mCRC between 2012 and 2022 at a high-complexity oncology center in Northeast Brazil. Molecular marker expression was evaluated by immunohistochemistry on TMA constructed from paraffin-embedded hepatic metastasis specimens. Disease-free survival (DFS) at 1, 3, and 5 years and overall survival (OS) were analyzed using Fisher's exact test, Kaplan-Meier curves, and Cox regression multivariate analysis.<h4>Results</h4>Median OS was 23 months (95% CI = 18.33-27.66). Among all molecular markers assessed, only pS6 positivity (20.7% of cases) showed a statistically significant association with worse DFS at 1 year (p = 0.032), 3 years (p = 0.038), and 5 years (p = 0.018). A significant association was also identified between pS6 positivity and FUS positivity (p = 0.014). In multivariate analysis for DFS, pS6 positivity was an independent predictor of recurrence at 3 years (HRa = 9.710; p = 0.028) and 5 years (HRa = 11.857; p = 0.018), as were postoperative complications at 3 years (HRa = 8.671; p = 0.014) and 5 years (HRa = 6.912; p = 0.029). For overall survival, three independent predictors of mortality were identified: adjuvant chemotherapy (HRa = 0.052; p < 0.001), absence of 1-year DFS (HRa = 3.641; p = 0.001), and pS6 positivity (HRa = 4.547; p = 0.001).<h4>Conclusion</h4>Median overall survival was 23 months, reflecting the high mortality burden of mCRC after hepatectomy. Among all molecular markers evaluated, pS6 was the only one independently associated with worse disease-free survival and higher mortality risk, consistent with its role as a surrogate marker of PI3K/AKT/mTORC1 pathway activation, though a direct causal relationship cannot be established from the present data. A significant association between pS6 and FUS positivity suggests a possible convergence of oncogenic pathways warranting further investigation. Postoperative complications were independent predictors of recurrence, and adjuvant chemotherapy was the strongest independent predictor of improved overall survival. Given the retrospective single-center design and limited sample size, these findings are hypothesis-generating and require prospective multicenter validation before clinical application.

Astrocytes, the most abundant glial cells in the central nervous system, play increasingly recognized roles in metabolic regulation beyond their classical supportive functions. This review summarizes current understanding of astrocyte morphological and molecular heterogeneity, with emphasis on regional metabolic properties across different brain areas. We discuss the primary forms and functional significance of astrocyte-neuron metabolic coupling, focusing on glucose, lipid, and glutamate metabolism, lactate shuttling, as well as the mitochondrial reactive oxygen species signaling. Furthermore, we integrate emerging evidence from animal and preclinical studies linking astrocyte-neuron metabolic coupling to ageing-related diseases. Collectively, these findings highlight how astrocyte-neuron interactions sustain physiological homeostasis and contribute to pathological manifestations, underscoring their significance in both health and ageing-related diseases.

Neurodegenerative diseases, including Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), and transmissible spongiform encephalopathies (TSEs), share fundamental mechanisms of protein misfolding, synaptic dysfunction, and progressive neuronal loss. The cellular prion protein (PrP^C), long viewed through the lens of prion biology, is now recognized as a pleiotropic modulator of neuronal physiology and a central interaction hub for amyloidogenic proteins. PrP^C undergoes tightly regulated proteolytic processing (α-, β-, γ-cleavage, and ectodomain shedding), generating soluble fragments and peptides with distinct and sometimes opposing biological activities. These derivatives modulate neurotrophic signaling, oxidative stress responses, and the uptake/toxicity of Alzheimer's disease-relevant soluble amyloid-β (Aβ), α‑synuclein, and tau. Recent work highlights PrP^C as a ligand for low-density lipoprotein receptor‑related protein‑1 (LRP1) and the NMDA receptor (NMDAR), with shed PrP^C and PrP‑derived peptides capable of initiating cell signaling and attenuating inflammatory responses, including when PrP^C is delivered via extracellular vesicles. In parallel, ultrasensitive seed amplification assays (SAA; RT‑QuIC/PMCA) have transformed prion diagnostics and underscore the biomarker potential of both full-length PrP^C and of its proteolytic fragments. Here, we synthesize current knowledge on PrP^C structure-function relationships, proteolytic processing, and crosstalk with disease‑ associated protein aggregates. We also outline emerging translational opportunities, ranging from PrP‑derived peptide therapeutics to fluid-based biomarkers that track disease onset and progression.

<h4>Background</h4>Hepatocellular carcinoma (HCC) is a genomically heterogeneous malignancy with substantial variability in prognosis and therapeutic response. Pseudouridine (<i>Ψ</i>) modification is an evolutionarily conserved RNA modification involved in RNA structure stabilization and translational regulation; however, the genomic and functional relevance of pseudouridine modification-related genes (PDGs) in HCC remains poorly defined.<h4>Methods</h4>Transcriptomic datasets and matched clinical information were collected from TCGA, GEO, and ICGC. Using feature-selection procedures and survival modeling, we established a pseudouridine-related risk signature. We next compared risk groups for genomic change, pathways, immune traits, and predicted drug response. Selected genes were tested in HCC cells by qPCR and CCK-8.<h4>Results</h4>The PDGs signature split HCC patients into high- and low-risk groups with different survival and remained prognostic after adjustment. The groups also differed in mutation, pathways, immunity, and chemotherapy response. DKC1 and PUS1 knockdown reduced proliferation.<h4>Conclusions</h4>This study identifies a PDGs signature linked with prognostic heterogeneity in HCC, supported by computational and experimental evidence.

The demineralization of tooth enamel is the primary consequence of dental caries, leading to cavities and finally tooth loss. Erosive tooth wear from acidic beverages and food is another factor that degrades enamel. In both cases, an acidic environment leads to etching and the final dissolution of tooth mineral, i.e., hydroxyapatite. Here, this process is discussed from a chemical perspective, taking into account the solubility of calcium phosphate and the presence of the pellicle (protein layer) and plaque (bacterial biofilms), which both affect the dissolution rate. While low pH is definitely decisive, calcium-binding ligands (e.g., acid anions, proteins) contribute to dissolution by removing calcium ions from the equilibrium. This is an important effect in the oral cavity where the concentration of biomolecules is high. The situation is complicated by the fact that the composition of saliva and the oral microbiome vary considerably between individuals. The state of current knowledge on the demineralization of enamel is summarized and discussed, also in the context of approaches to prevent dental caries and erosive tooth wear.

<b>Background:</b> Atrial fibrillation (AF) is the most common sustained cardiac arrhythmia and is strongly associated with atrial structural remodeling driven by activated cardiac fibroblasts. Autophagy has been implicated in AF-related atrial remodeling; however, the non-coding RNA mechanisms that govern autophagic activation in atrial fibroblasts under rapid electrical stress remain poorly understood. <b>Methods:</b> Human cardiac fibroblasts from adult atria (HCF-aa) were subjected to rapid electrical stimulation (RES) at 0.5 V/cm and 10 Hz. Expression levels of exosomal metastasis-associated lung adenocarcinoma transcript 1 (MALAT1), cytoplasmic miR-204-5p, and microtubule-associated protein light chain 3B (LC3B) were measured using quantitative real-time PCR and Western blot analyses. Luciferase reporter assays were performed to confirm direct molecular interactions. The functional roles of MALAT1 siRNA, miR-204-5p mimics/antagomirs, rapamycin, and 3-methyladenine (3-MA) on LC3B expression and autophagic activation were assessed by Western blot and immunofluorescence confocal microscopy for LC3B puncta formation. <b>Results:</b> RES significantly induced exosomal MALAT1 expression in a voltage- and time-dependent manner, peaking at 2 h post-stimulation, while cytoplasmic MALAT1 levels remained unchanged. Cytoplasmic miR-204-5p exhibited an initial transient rise followed by a significant decline at 2 h, inversely correlating with peak MALAT1 levels. LC3B mRNA and protein expression subsequently increased, peaking at 6 and 16 h, respectively. Luciferase reporter assays confirmed that miR-204-5p directly binds both the MALAT1 transcript and the 3'-UTR of LC3B mRNA. MALAT1 knockdown augmented miR-204-5p levels and suppressed LC3B expression, while miR-204-5p overexpression attenuated RES-induced LC3B upregulation and LC3B puncta accumulation. Conversely, miR-204-5p inhibition further enhanced autophagic activation, as evidenced by increased LC3B puncta density. <b>Conclusions:</b> In HCF-aa subjected to RES, MALAT1 functions intracellularly as a competing endogenous RNA to putatively sequester miR-204-5p, thereby de-repressing LC3B expression and promoting autophagic activation. Concurrent exosomal secretion of MALAT1 may additionally serve as a paracrine signal to neighboring cells, though this requires future conditioned-media transfer experiments to confirm.

Cardiac regeneration strategies increasingly leverage transient reprogramming to restore function in damaged myocardium. Here, we show that delivery of modRNA encoding STEMIN, a modified serum response factor, together with YAP5SA, a constitutively active YAP1 variant, induces coordinated chromatin remodeling and transcriptional reprogramming in cardiomyocytes. Using ATAC-seq and RNA-seq in rat and human cardiomyocytes, we identify the activation of cell cycle, DNA replication, and survival-associated pathways, alongside a robust induction of microRNAs linked to apoptosis resistance. Exosome profiling reveals selective packaging and release of these miRNAs, which may target key regulators of intrinsic and extrinsic apoptotic pathways, including caspases and TP53-associated signaling. Functional assays demonstrate reduced apoptosis <i>in vitro</i> and <i>in vivo</i>. These findings support a model in which reprogramming factor-induced exosomal signaling enhances cardiomyocyte survival by inhibiting apoptosis, and highlight a potential therapeutic framework that integrates modRNA delivery with paracrine mechanisms for cardiac repair.

β-Tricalcium phosphate (β-TCP, Ca₃(PO₄)₂) is a widely used bioresorbable ceramic in orthopedic applications, particularly as a bone graft material and as a coating on bioinert metallic implants. Its bioactivity, resorption kinetics, and biocompatibility account for its extensive clinical use, motivating efforts to further enhance its performance. The incorporation of various ions into the β-TCP lattice has been widely explored to tailor its structural, physicochemical, and biological properties. Among these, selenium (Se) is an essential trace element that enhances osteoblast activity and supports bone regeneration. To achieve this, microwave-assisted synthesis was employed for its advantages, including rapid reaction kinetics, homogeneous heating, limited grain growth, and efficient ion incorporation. In this study, submicron-sized selenite (SeO₃ ^2-)-doped β-TCP (Se-β-TCP) powders were successfully synthesized using this technique and characterized in detail. Structural analyses confirmed the incorporation of SeO₃ ^2- into the β-TCP lattice, as evidenced by a reduction in the c parameter from 37.372 Å to 37.242 Å and the emergence of a Se-O vibration band at 904-806 cm^-1, in the infrared spectra. In vitro results showed that MC3T3-E1 preosteoblasts cultured with Se-β-TCP proliferated for up to 5 days, while the powders inhibited colony growth of <i>Staphylococcus aureus</i>, <i>Staphylococcus epidermidis</i>, and <i>Escherichia coli</i>. These findings demonstrate that Se-β-TCP synthesized via microwave-assisted processing is both cytocompatible and antibacterial, highlighting its potential for orthopedic applications.

<h4>Objective</h4>Sepsis-associated acute kidney injury (SA-AKI) is a critical complication that substantially increases intensive care unit mortality. Early identification is paramount for timely intervention. This study aimed to develop and internally validate a prediction model relying exclusively on the first serum laboratory indicators after hospital admission to predict SA-AKI risk at the earliest available laboratory assessment.<h4>Methods</h4>Clinical data of 1,573 sepsis patients admitted to West China Hospital of Sichuan University (January 2024-December 2025) were retrospectively analyzed. Patients were divided into SA-AKI and non-SA-AKI groups per 2012 KDIGO criteria, and randomly split into training (70%, <i>n</i> = 1,102) and validation (30%, <i>n</i> = 471) cohorts. LASSO regression screened potential predictors exclusively from first serum laboratory indicators after admission, and multivariate logistic regression constructed the model. A nomogram was constructed to facilitate individualized risk estimation. Subgroup analyses were performed across age and sex strata. Model performance was evaluated via ROC curves, calibration curves, and decision curve analysis (DCA).<h4>Results</h4>LASSO regression selected 10 serum indicators, and multivariate logistic regression confirmed 8 independent risk factors: myoglobin (MYO), alanine aminotransferase (ALT), phosphorus (PO₄), sodium (Na), potassium (K), carbon dioxide combining power (CO₂-CP), platelet (PLT), and neutrophil (NEUT) (all <i>P</i> < 0.05). The model exhibited favorable discrimination (training cohort <i>AUC</i> = 0.839 [95% <i>CI</i>: 0.814-0.864]; validation cohort <i>AUC</i> = 0.832 [95% <i>CI</i>: 0.791-0.874]), acceptable graphical calibration, and significant clinical utility across a wide threshold probability range. Subgroup analyses showed stable discriminative performance across age and sex strata (all AUCs > 0.81).<h4>Conclusions</h4>This study developed and internally validated a promising predictive model for estimating SA-AKI risk in sepsis patients using solely first routine serum laboratory indicators after admission. A nomogram is provided for individualized bedside risk estimation. This tool may support early risk stratification of high-risk individuals. External validation in multi-center, diverse cohorts is warranted before broader clinical implementation.

<h4>Background</h4>Metabolic dysfunction-associated steatotic liver disease (MASLD) is increasingly recognized as a contributor to hepatocellular carcinoma. Whether repeated assessment of MASLD provides additional prognostic information for liver cancer risk remains unclear.<h4>Methods</h4>Using the UK Biobank, we included 12,111 participants who underwent both baseline and follow-up health screenings. MASLD was defined at each assessment as fatty liver index ≥60 plus at least one cardiometabolic risk criterion. Four trajectory groups were defined: no MASLD → no MASLD, no MASLD → MASLD, MASLD → no MASLD, and MASLD → MASLD. Cox proportional hazards models were used to estimate adjusted hazard ratios (aHR) and 95% confidence intervals (CI).<h4>Results</h4>At baseline, 4,008 participants (33.1%) had MASLD. MASLD at period 1 was associated with a higher risk of incident liver cancer compared with no MASLD (aHR, 2.81; 95% CI, 1.20-6.58). A similar association was observed for MASLD at period 2 (aHR, 2.61; 95% CI, 1.12-6.06). In trajectory analyses, MASLD → no MASLD (aHR, 4.00; 95% CI, 1.01-15.80) and MASLD → MASLD (aHR, 3.51; 95% CI, 1.31-9.44) were associated with higher liver cancer risk than no MASLD → no MASLD.<h4>Conclusions</h4>MASLD at either assessment was associated with higher incident liver cancer risk. Trajectory analyses suggest that persistent MASLD and prior MASLD exposure may both carry prognostic importance.

Network physiology addresses global physiological states that arise self-organized through the interactions among the individual components of the human organism and is therefore intimately linked to Synergetics with its central concepts of order parameters, enslavement, and circular causality. Explanations of self-organization phenomena invoking these concepts exhibit a distinctive explanatory power and scientific fruitfulness across disciplines, in part due to their appeal to downward causation. Downward causation is not only actively discussed in philosophy and philosophy of science, particularly in debates on reductionism and emergence, but is also attracting considerable interest in scientific research on complex systems. However, at least when understood in ontological terms, it remains a philosophically delicate and contested notion.

The Solute Carrier family 27 (SLC27), also known as the fatty acid transport proteins (FATPs) is a group of transmembrane integral proteins that facilitate the entry of long-chain and very long-chain fatty acids into cells. The human SLC27 family consists of six highly homologous members (SLC27A1 to A6), which are expressed in various tissues reliant on fatty acids for energy metabolism and biosynthesis, participating in multiple physiological processes. Recent years have seen extensive research on the role of SLC27 in lipid metabolism and more recently in the exploration of the family in the initiation and progression of cancer. Aberrant expression of SLC27s in cancers have been reported, suggesting their potential involvement in key processes such as tumour growth, invasion, metastasis, and immune evasion by promoting fatty acid uptake, remodeling cellular metabolic networks, activating lipid-related signaling pathways, and even modulating the tumour immune microenvironment. Given their significant role in tumor metabolic reprogramming, SLC27s are increasingly regarded as potential therapeutic targets for cancer, attracting widespread attention. The present article summarises the basic characteristics and regulatory mechanisms of the SLC27 family, as well as their cellular functions and roles in cancer, exploring the potential of these family members in tumour metabolism and therapy, thereby providing new directions for future clinical research.

Persistent transaminitis has a very broad differential diagnosis, and extrahepatic causes may often be initially overlooked. Anti-3-hydroxy-3-methylglutaryl-CoA reductase (anti-HMGCR) myopathy is a subtype of immune-mediated necrotizing myopathy (IMNM) characterized by the presence of anti-HMGCR antibodies and myofiber injury. Patients typically present with proximal extremity weakness, transaminitis, and a history of statin use. Unlike statin intolerance or statin myopathy, the weakness and biochemical abnormalities persist despite discontinuation of the drug. Here, we present a case of a patient with transaminitis and elevated creatine kinase (CK) levels, which persisted after cessation of his statin and were found to be associated with mildly elevated anti-HMGCR antibodies. Interestingly, he did not require immunosuppression for the resolution of his myopathy.

<h4>Background</h4>Ménière disease (MD) is a chronic inner ear disorder characterized by recurrent vertigo, fluctuating sensorineural hearing loss, tinnitus, and aural fullness. Diagnosis is clinical, and no validated peripheral biomarkers are available. Although endolymphatic hydrops is a hallmark, its poor correlation with symptom severity suggests broader systemic mechanisms.<h4>Methods</h4>We performed large-scale serum proteomic profiling using the Olink Explore HT platform, quantifying 5400 proteins in patients with definite MD and matched controls. After quality control, 1037 proteins were analyzed through differential expression, protein-protein interaction mapping, and systems biology approaches.<h4>Results</h4>MD exhibited a structured systemic proteomic signature characterized by 3 functionally integrated protein families in the upregulated proteome: (1) neuroendocrine and fluid-regulatory mediators (NPPB, POMC, CCK, NPY) converging on pathways controlling fluid homeostasis and HPA axis signaling; (2) morphogenetic and tissue-remodeling proteins (BMP6, SOST, FRZB, RSPO/SFRP) supporting extracellular matrix plasticity; and (3) cellular stress-adaptation proteins (FARSA, WARS1, BAG3) involved in translational control and proteostasis. The downregulated proteome revealed coordinated suppression of epithelial maintenance factors (EGF, SBDS) and neutrophil immune effectors (MMP8, OLFM4, RNASE3) alongside attenuation of NF-κB signaling, indicating dysregulation of tissue repair and immune surveillance rather than hyperinflammation. Tissue enrichment highlighted autonomic and vascular compartments in the upregulated signature, while neutrophil and hematopoietic lineages dominated the downregulated profile, supporting a neurovascular-immune interface.<h4>Conclusions</h4>MD emerges as a systemic neurovascular-immune disorder characterized by activation of compensatory neuroendocrine-vascular-proteostatic pathways coupled with dysregulation of tissue repair and immune effector mechanisms, rather than solely a hydropic condition. The identified serum proteomic signature provides a candidate biomarker framework for diagnostic support, patient stratification, and future longitudinal validation.

The Enroll-HD natural history platform provides rich longitudinal phenotypes enabling genome-wide analyses across diverse clinical domains. Psychiatric symptoms are a major source of morbidity in Huntington’s disease (HD), yet the genetic architecture underlying their onset is poorly understood. We analyzed ∼18,000 people with HD (PwHD) to define genetic determinants of ages at psychiatric, motor, and cognitive symptom onset, and HD diagnosis. GWAS meta-analysis recapitulated 11 established modifiers of motor onset and identified a novel locus spanning RAB3B / ZFYVE9 associated with age at violent/aggressive behavior onset. Exome-wide analyses in Enroll-HD participants implicated rare variants in FAN1 , PMS1 , POLD1 , and HTT . Several HD modifiers of motor and cognitive symptom onset ( MSH3 , FAN1 , HTT ) also influenced psychiatric symptom onset, whereas PMS1 and POLD1 showed significant association with motor symptom onset. Psychiatric polygenic scores predicted psychiatric symptom onset, revealing a hybrid architecture combining psychiatric liability in general population with HD- or repeat expansion disease (RED)-specific pathways.

<h4>ABSTRACT</h4> Huntington’s disease is a rare neurodegenerative disease whose primary risk factors are inherited expansions of a CAG repeat tract in the HTT gene. Somatic expansion of these tracts leads to neuronal toxicity, neuronal death and clinical disease progression. To identify genetic factors with a major impact on disease onset and progression, we genome sequenced 18,825 individuals for the ENROLL-HD study. Our results show rare inactivating mutations in three genes, all involved in DNA damage repair, are major determinants of age of onset for motor symptoms (n=10,610) and other clinical manifestations. Heterozygote carriers of predicted loss-of-function (pLoF) variants in POLD1 and PMS1 developed motor symptoms an average 20 years (n=3; P=1×10 −5 ) and 7 years (n=6; P=2×10 −3 ) later than non-carriers, respectively. Conversely, heterozygote carriers of pLoF variants in FAN1 (n=30) developed symptoms 10 years earlier (P=2×10 −10 ). Our findings highlight therapeutic strategies and help predict age of onset for at-risk individuals.

Expanded short tandem repeats contribute to a broad spectrum of neurodegenerative diseases, yet their roles in Parkinson’s disease (PD) and parkinsonism remain incompletely characterized, especially across diverse ancestries. We analyzed short-read whole-genome (WGS) and clinical exome sequencing (CES) data from 38,365 individuals (28,861 WGS; 9,504 CES), encompassing 23,242 patients with PD, 4,729 patients with atypical parkinsonism and 10,394 healthy controls from 11 genetic ancestries. To determine carrier frequencies and characterize repeat structures across diverse ancestries, we genotyped 12 established pathogenic loci where normal, intermediate, and pathogenic alleles can be reliably differentiated using short-read sequencing data. Additionally, we conducted threshold-based associations to determine the minimum threshold associated with increased PD risk in 15,995 individuals (8,591 PD, 7,404 controls) of European ancestry. Pathogenic repeat expansions were detected in 62 patients (56 PD and 6 atypical parkinsonism) and 5 controls across seven loci ( AR , ATXN1 , ATXN2 , ATXN3 , CACNA1A , HTT and THAP11 ), spanning seven ancestries. Among these, ATXN2 expansions were the most frequently observed in PD and were present in African, East Asian, European and Middle Eastern ancestries. Additionally, intermediate ATXN2 repeat expansions exhibited a strong, length-dependent association with PD risk in the European population, with individuals with ≥32 repeats having a more than four-fold increased risk (odds ratio 4.25, 95% confidence interval 1.80-12.05). Overall, >92% of expanded alleles harbor CAA interruptions within the CAG tract. Pathogenic expansions at other loci, such as ATXN3 and THAP11 , showed more ancestry-specific distributions. Clinically, individuals with pathogenic ATXN2 and ATXN3 expansions most often presented with typical PD features but frequently showed earlier disease onset and a strong family history of PD. This large-scale, multi-ancestry study comprehensively maps the genetic landscape of pathogenic and intermediate repeat expansions in PD. Our findings confirm a length- and structure-dependent risk association for ATXN2 with PD in the European population and highlight the pleiotropic effects of repeat expansions across the parkinsonian spectrum.

Alzheimer's disease (AD) is a progressive neurodegenerative disorder marked by extracellular amyloid-β (Aβ) plaques, intracellular neurofibrillary tangles of hyperphosphorylated tau, synaptic dysfunction, and chronic neuroinflammation. AD pathogenesis involves multiple central nervous system (CNS) cell types-including neurons, astrocytes, microglia, and oligodendrocytes, and, less prominently, neural stem cells (NSCs), ependymal cells, and endothelial cells-which undergo coordinated but cell-type-specific pathological changes. These include neuronal loss, reactive gliosis, impaired myelin maintenance, reduced neurogenesis, and blood-brain barrier (BBB) dysfunction. MicroRNAs (miRNAs), the small non-coding RNAs that regulate post-transcriptional gene expression, have emerged as key modulators of these cell-specific processes and are consistently dysregulated in AD. Across AD-vulnerable brain regions and CNS cell types, miRNAs influence amyloid and tau biology, synaptic resilience, glial activation states, myelin structure, neurogenic potential, and vascular homeostasis. Dysregulated miRNAs also act across cell types through extracellular vesicle (EV) transfer, amplifying or mitigating amyloidogenesis, tauopathy, neuroinflammation, and white-matter injury. This review provides a comprehensive, cell-type-specific analysis of miRNAs involved in AD, detailing their roles in neurons, astrocytes, microglia, oligodendrocytes, NSCs, ependymal cells, and endothelial cells. We highlight common miRNAs that function across multiple CNS cell types and examine the potential of circulating and cerebrospinal fluid (CSF) miRNAs as minimally invasive biomarkers. Finally, we discuss therapeutic strategies aimed at restoring protective miRNAs or inhibiting pathogenic miRNAs, emphasizing the need for targeted interventions. By integrating pathways of miRNA dysregulation across CNS cell types, this review underscores the central role of miRNA networks in AD pathogenesis and the promise of precise, cell-specific miRNA modulation.

Senescent cells accumulate in chronically diseased liver tissues and are known to actively contribute to disease pathology. To date, these studies have predominantly focussed on senescence in epithelial cells, such as hepatocytes and biliary epithelial cells, and senescence in liver endothelial cells remains largely understudied. Here, we utilise publicly available single-cell RNA-sequencing data, immunohistochemical and immunofluorescent staining to detect senescent endothelial cells within chronically diseased human liver tissues. Next, we develop a novel protocol for the induction of paracrine senescence in primary human liver endothelial cells and explore their functionality. We demonstrate that senescent liver endothelial cells exhibit a reduced scavenging capacity but are still able to support lymphocyte recruitment under physiological flow conditions in vitro. Mechanistically, we determine that inducible T cell costimulator ligand (ICOSL) is an important factor in the specific recruitment of CD4⁺ T cells, but antibody blockade, genetic knockdown and genetic overexpression of ICOSL in endothelial cells has no effect on CD8⁺ T cell recruitment. Finally, we show that ICOSL gene expression is upregulated in chronically diseased tissues, present in scar-associated endothelial cells and correlates to CD4⁺ T cell infiltration. This is the first study to demonstrate that senescent human liver endothelial cells can potentially shape the liver immune microenvironment in chronic liver disease. Targeting senescent endothelial cells could present new therapeutic opportunities to treat chronic liver diseases.

<h4>Background</h4>Transcript-level analyses allow for the precise characterization of gene expression and its functional role in cancer. However, most of these studies rely on reanalyses of next-generation sequencing data, whose incomplete or inaccurate assemblies limit the comprehensive and faithful characterization of transcripts. To systematically elucidate transcriptomic expression in tumors and define broadly applicable therapeutic strategies, we investigated cancer-specific RNA transcripts (cancer-SRTs) expressed across multiple cancer types based on long-read sequencing data.<h4>Methods</h4>We characterized the expression profiles of 44,405 cancer-SRTs across multiple cancer types using t-SNE and correlation analyses. Transcripts expressed in more than 10 cancer types were further investigated through enrichment, survival, and correlation analyses to elucidate their functions and clinical relevance. To explore the mechanisms driving cancer-SRT generation, we analyzed alternative splicing events within these transcripts and integrated copy number variation, DNA methylation, and ATAC-seq data from matched TCGA tumor samples. Using the expression of 131 transcripts strongly associated with tumor hallmarks, we developed a risk-score model to evaluate associations with patient survival, tumor stage, immune characteristics, and responses to immune checkpoint blockade. Finally, the in vitro anti-tumor effects of siRNAs targeting two cancer-SRTs were evaluated using CCK-8 assay, colony formation, and transwell assays.<h4>Results</h4>Cancer-SRTs exhibit substantial structural diversity and are enriched in malignancy-associated pathways. The expression of these transcripts is associated with multiple genomic and epigenetic processes. We identify 131 transcripts that are strongly associated with tumor hallmarks and develop a risk-score model for evaluating patient prognosis and tumor progression. The model also exhibited strong associations with features of immune evasion.<h4>Conclusions</h4>Cancer-SRTs are widely expressed yet highly heterogeneous across tumor types, and are subject to multiple regulatory mechanisms underlying their functional and clinical significance. These findings advance our understanding of tumor biology and lay the groundwork for developing diagnostic, prognostic, and therapeutic strategies based on these transcripts. Future studies investigating their underlying mechanisms and applications in immunotherapy will be critical for precision cancer treatment.

<h4>Background</h4>Dementia with Lewy bodies (DLB) is a multisystem neurodegenerative disorder often accompanied by autonomic and neuropsychiatric symptoms. Nutritional impairment is increasingly recognized yet under-characterized in DLB. DLB was chosen due to its early multisystem involvement, including brainstem-mediated autonomic and sleep disturbances, which may uniquely affect nutrition. This study evaluated nutritional status and its associations with cognitive, mood, autonomic, and sleep-related features.<h4>Methods</h4>Seventy patients with probable DLB (2017 McKeith criteria) were retrospectively included. Cases of Parkinson's disease dementia or other Lewy body-related conditions were excluded. Nutritional status was assessed with the Mini Nutritional Assessment-Short Form (MNA-SF). Cognitive function (Addenbrooke's Cognitive Examination, ACE-III), depressive symptoms (Geriatric Depression Scale, GDS), motor severity (Hoehn and Yahr staging), sleep disturbances (Rapid Eye Movement Sleep Behavior Disorder, RBD, insomnia), and autonomic symptoms (urinary incontinence, constipation, orthostatic hypotension) were extracted from clinical records. Multivariable logistic, ordinal regression, and hierarchical cluster analyses were performed.<h4>Results</h4>Normal nutrition was observed in 14.3% of patients, 60.0% were at risk, and 25.7% were malnourished. Malnutrition was significantly associated with lower ACE-III and higher GDS scores (p < 0.05). RBD and urinary incontinence differed across nutritional groups. Ordinal regression showed age and female sex independently associated with worse nutritional status. Cluster analysis identified a high-risk phenotype with greater cognitive, depressive, autonomic, and sleep-related burden linked to malnutrition.<h4>Conclusion</h4>Malnutrition is highly prevalent in DLB and correlates with cognitive, psychiatric, autonomic, and sleep-related features. These associations support that nutritional impairment reflects the multisystem disease burden. Early identification of patients with combined deficits may help target those at risk and guide comprehensive management.

Opioid use disorder (OUD) involves separable stages of exposure and dependence, but many genetic studies have treated these liabilities as a single phenotype. In this study, transcriptome-wide association study (TWAS) methods were applied to the Psychiatric Genomics Consortium Substance Use Disorders 2020 opioid meta-analysis summary statistics to examine ageing-related gene pathways across brain tissues. Genetic liability for opioid exposure was strongly associated with reduced predicted expression of complement components and synaptic-pruning genes, together with negative signals in astrocyte and senescence-related gene sets. By contrast, liability for dependence among opioid-exposed individuals showed positive enrichment in AMP-activated protein kinase-mechanistic target of rapamycin nutrient-sensing pathways, mitochondrial oxidative phosphorylation, especially DLD, and selected nicotinamide adenine dinucleotide/sirtuin pathways, accompanied by presynaptic and DNA repair-associated signals. Many ageing-related gene sets showed opposing directional profiles between exposure and dependence contrasts, indicating that these stages are associated with partially antagonistic transcriptomic profiles. Gene-level analyses identified influential drivers and directional sign-flips, including CASP7, TERF2, STX1A, and ADCY6, which further supported the stage-specific pattern. A dedicated post-hoc robustness inventory was performed, including threshold variation, family-wise false discovery rate pooling, tissue-specific decomposition, gene-set overlap analysis, major histocompatibility complex/complement flagging, and leave-one-out influence analysis. In this inventory, 131 of 4913 pooled statistical tests survived global Benjamini-Hochberg false discovery rate correction, and 100 of 102 evaluable pathway-profile entries satisfied an opposing exposure-versus-dependence criterion. These findings suggest that opioid exposure and opioid dependence are biologically distinguishable at the level of ageing-related brain gene expression. The results nominate stage-specific risk modules and biomarker hypotheses that may inform future precision approaches to OUD prevention and treatment while remaining hypothesis-generating until validated in functional and longitudinal studies.

<b>Background/Objectives</b>: The transcription factor carbohydrate response element-binding protein (ChREBP) is a key glucose-sensing regulator that governs glucose and lipid metabolic homeostasis. However, its specific functions in skeletal muscle remain insufficiently clarified. The present study aimed to investigate the roles of ChREBP in skeletal muscle exercise capacity, energy metabolism, and adaptive remodeling, as well as muscle regeneration. <b>Methods</b>: We generated a skeletal muscle-specific ChREBP knockout mouse model, and assessed their exercise performance, energy metabolism, skeletal muscle fiber composition, and injury repair capacity. Additionally, hypoxia and high-fructose diet models were established to analyze the function of ChREBP in skeletal muscle adaptive remodeling. C2C12 myoblasts and primary muscle satellite cells were used to explore its effects on myogenic differentiation. <b>Results</b>: Genetic deletion of ChREBP induced no detectable alterations in myofiber composition, overall metabolic status, or muscle adaptive remodeling triggered by hypoxia and high-fructose diet. In vitro assays demonstrated that ChREBP overexpression facilitates C2C12 myogenic differentiation. Adeno-associated virus-mediated ChREBP overexpression enhanced histological markers of regeneration, including desmin-positive regenerative area and the cross-sectional area of newly formed myofibers after cardiotoxin-induced injury. <b>Conclusions</b>: Collectively, our experimental data indicate that ChREBP is largely dispensable for maintaining basal skeletal muscle homeostasis and stress-induced adaptive remodeling. Meanwhile, this study identifies a previously unrecognized regulatory role of ChREBP in the processes of skeletal muscle damage repair and post-injury regeneration.

Self-healing biomaterials have attracted significant attention due to their ability to restore structural integrity, extend material lifetime, and reduce maintenance costs without external intervention. In this study, Polyvinyl Alcohol/Graphene Oxide/Eggshell Particle (PVA/GO/ESP) composite hydrogels were synthesized via a freeze-thawing method and characterized using XRD, SEM/EDS, and FTIR analyses. The effect of ESP incorporation on the self-healing and mechanical properties of the hydrogels was systematically investigated. Tensile test results demonstrated that incorporation of 1 wt% ESP improved the tensile strength up to 0.326 MPa while maintaining high strain capacity. Healing efficiency values calculated from recovered tensile strength showed approximately 69%, 47%, and 67% recovery for PVA/GO, PVA/GO/ESP (0.5%), and PVA/GO/ESP (1%) hydrogels, respectively. The developed hydrogels demonstrated rapid self-healing behavior at room temperature without external stimuli. These findings suggest that ESP-reinforced PVA/GO hydrogels may serve as promising candidates for future biomaterial and soft tissue engineering studies. The developed hydrogels demonstrated enhanced tensile strength, rapid self-healing behavior, and promising swelling properties, indicating their potential use in soft tissue engineering and biomaterial applications.

<h4>Background</h4>Renal fibrosis is a progressive injury contributing to renal function deterioration. Mounting evidence has underscored the profound impact of gut microbiota metabolites on host health and disease, yet their underlying mechanisms against renal fibrosis remain unclear. The aim of this study was to fully elucidate their therapeutic potential in the context of renal fibrosis.<h4>Methods</h4>The targets of gut microbiota metabolites were identified in gutMGene. The diseases targets were obtained from the OMIM, GeneCards and DisGeNet databases. The STRING and DAVID platform were employed to identify the core targets and pathways. Gut Microbiota-Targets-Pathway-Metabolites (G-T-P-M) network was constructed to screen the core metabolites. Molecular docking was used to assess the interactions between the targets and metabolites.<h4>Results</h4>A total of 47 overlapping targets related to gut microbiota metabolites and renal fibrosis were acquired. The bioinformatics analysis indicated that the targets were enriched in the regulation of TNF pathway and Toll-like receptor pathway. The PPI network (Protein-Protein Interaction) identified JUN, IL6, IL1B and AKT1 as the core targets. The G-T-P-M network revealed that Propionate, Butyrate and 3-Indolepropionic acid were identified as the core non-toxic and promising core metabolites. The core metabolites showed stable binding affinity with the core targets.<h4>Conclusion</h4>The findings highlight that gut microbiota metabolites represent a promising therapeutic option for combating renal fibrosis by modulating multiple targets and pathways, providing a theoretical foundation for the future studies exploring gut microbiota as targeted strategies in the prevention and treatment of renal fibrosis.

Raf Kinase Inhibitory Protein (RKIP), also known as PEBP1, is a multifunctional modulator of intracellular signaling with pivotal roles in cellular homeostasis, cancer suppression, cardiac physiology, and neurobiology. Initially characterized by its inhibition of the Raf-1/MEK/ERK cascade, RKIP has since emerged as a dynamic regulator of numerous pathways, including NF-κB, GRK2, GSK3β, and Aurora B kinase. RKIP activity is modulated through phosphorylation-dependent conformational shifts that dictate its binding partners and regulatory outcomes. In oncology, RKIP acts as a metastasis suppressor by promoting let-7 microRNA expression and inhibiting pro-metastatic genes such as HMGA2, BACH1, MMPs, and CXCR4. In the nervous system, RKIP influences synaptic signaling, pain perception, and neuroprotection, while in cardiomyocytes, it enhances β-adrenergic signaling and protects mitochondria under stress. Dysregulation of RKIP is implicated in cancer progression, heart failure, and neurodegenerative diseases. Ongoing research into pharmacological modulation of RKIP holds promise for novel therapeutic interventions across diverse pathologies.

<h4>Background</h4>Vascular cognitive impairment (VCI) is a devastating clinical endpoint of microvascular senescence. However, the mechanisms by which age-related focal vascular insults cause systemic brain network failure and molecular vulnerability remain unknown. To decode the multi-scale neurobiology of VCI, we conducted a systematic review and meta-analysis of whole-brain voxel-based morphometry studies comparing patients with VCI and healthy controls.<h4>Methods</h4>We used coordinate-based network mapping on a normative functional connectome to identify convergent structural atrophy networks. To decode multi-scale biological substrates, we checked the resulting macroscopic topography against the Allen Human Brain Atlas and 28 positron emission tomography-derived neurotransmitter maps.<h4>Results</h4>18 studies contributed to the analysis, including 682 VCI patients and 643 healthy controls. VCI-related atrophy, despite appearing disparate, functionally converges onto a robust macroscopic architecture that is anchored predominantly in the somatomotor and salience networks. Transcriptomic profiling further showed that this network colocalizes significantly with Layer 6 corticothalamic and subcortical projection neurons. These neuron populations feature exceptionally long axonal projections, a property that heightens their metabolic susceptibility to chronic hypoperfusion. At the neurochemical level, this structural degradation exhibited profound spatial coherence with the macroscopic distribution of dopamine transporter (DAT) and 5-hydroxytryptamine transporter (5‑HTT).<h4>Conclusion</h4>These findings suggest that VCI may represent a quintessential "disconnection syndrome" associated with the vulnerability of long-range projection pathways to vascular aging, providing a novel multi-scale neurobiological template to identify network-level targets for intervention.

Studies on the mechanosensitive ion channel Piezo2 largely focus on its role in the peripheral nervous system, particularly in touch and pain sensation. Here, we investigate Piezo2 function in the anterior visual pathway of the central nervous system with a focus on oligodendrocyte (OL) biology and myelin integrity. Using single-nucleus RNA sequencing, we identify Piezo2 expression in late differentiated OLs of the murine optic nerve, with minor expression in retinal ganglion cells. OL-specific Piezo2 deficiency results in age-dependent motor impairment and selective disruption of myelin compaction in small-caliber optic nerve axons, a fiber population known to be particularly vulnerable in demyelinating disease. Differential gene expression analysis further indicates that Piezo2 regulates myelin compaction and white matter integrity in mature OLs. Consistent with these findings, OL-encoded PIEZO2 expression is reduced in optic nerve lesion areas from multiple sclerosis patients, highlighting a convergent mechanism of small-caliber fiber vulnerability. Together, these data identify Piezo2 as an age-related regulator of OL function and myelin integrity, with potential relevance for preserving white matter structure in multiple sclerosis.

<h4>Background</h4>Emerging evidence shows that morning chronotype is associated with a reduced risk of breast cancer, yet biological mechanisms remain unclear. This study aimed to identify circulating proteins that mediate this association.<h4>Method</h4>We employed a two-step Mendelian randomization approach using European-ancestry genome-wide association studies. Exposure data on self-reported morning chronotype were from the UK Biobank and 23andMe (372,765 cases, 278,530 controls). Mediators included 4907 plasma proteins measured by SomaScan in 35,559 Icelandic individuals. Outcome data for overall breast cancer and subtypes were sourced from the Breast Cancer Association Consortium (133,384 cases, 113,789 controls) and FinnGen DF 11 (20,586 cases, 201,494 controls). We identified mediators through concordant step 1 and step 2 associations, followed by colocalization (posterior probability > 0.80) and mediation analyses. Single-cell RNA sequencing in human breast cancer tissues was used to assess biological relevance.<h4>Results</h4>Genetically predicted morning chronotype is associated with a lower risk of breast cancer (odds ratio, 0.93; 95% confidence interval, 0.89 to 0.98). Morning chronotype is associated with 895 plasma proteins, of which seven proteins show inverse associations with overall breast cancer (false discovery rate <0.05). Colocalization support five mediators sharing causal variants for protein levels and breast cancer susceptibility. Mediation and single-cell RNA sequencing analyses further confirm their biological roles.<h4>Discussion</h4>These findings uncover plausible biological pathways linking morning chronotype to reduced breast cancer risk, nominating ADAM15, BTN2A1, CASP8, PDCD6, and RSPO3 as potential therapeutic targets. This work bridges epidemiological observations with mechanistic insights, offering a translational roadmap for chronotype-based interventions.

<h4>Background</h4>Methyl jasmonate (MeJA) plays a key role in enhancing plant resistance to a wide range of diseases. However, the specific defense responses it induced and the underlying molecular mechanisms remain incompletely understood in many plant-pathogen systems. This study aimed to determine whether exogenous MeJA enhances resistance to leaf spot disease caused by Alternaria tenuissima in peony (Paeonia lactiflora) and to elucidate the associated molecular and biochemical mechanisms through integrated transcriptomic and biochemical analyses.<h4>Results</h4>Compared with the control (CK), pretreatment with 25-75 µmol L^- 1 MeJA significantly reduced disease severity, with 75 µmol L^- 1 identified as the optimal concentration. Peony leaves pretreated with this concentration exhibited a stronger hypersensitive response than the CK, with initial differentiation observed at 24 h after inoculation. Transcriptome sequencing analysis identified 1,017 differentially expressed genes between the MeJA-pretreated and CK groups, primarily associated with cell wall organization, phenylpropanoid biosynthesis, terpenoid backbone biosynthesis, and flavonoid biosynthesis pathways. MeJA pretreatment up-regulated the genes involved in phenylpropanoid and lignin biosynthesis, including peroxidase 27 (POD27) and caffeic acid 3-O-methyltransferase (COMT), as well as flavonoid-related genes such as chalcone isomerase (CHI). Genes regulating pectin methylesterification, including pectin methylesterase (PME) and PME inhibitor (PMEI), were also modulated. In addition, transcription factors such as basic leucine zipper (bZIP) and heat shock transcription factor (HSF) were activated. Biochemical analyses confirmed increased levels of major cell wall components (lignin, cellulose, hemicellulose, and pectin) and total flavonoids were significantly higher in MeJA-pretreated plants compared with CK plants. A putative model of MeJA-induced defense responses against A. tenuissima was proposed.<h4>Conclusions</h4>Exogenous MeJA pretreatment enhances resistance to leaf spot disease caused by A. tenuissima in peony by promoting cell wall reinforcement and flavonoid accumulation. These findings highlight the potential of MeJA as a strategy for managing leaf spot disease in peony cultivation.

Following injury, prostaglandin E₂ (PGE₂) drives intestinal epithelial repair by inducing revival stem cells (RSCs), which compensate for the loss of homeostatic Lgr5⁺ stem cells. Using intestinal organoid models, we demonstrate that melatonin potentiates the PGE₂- or damage-induced RSC emergence by rewiring cellular plasticity toward a fetal-like state and sustaining pro-regenerative YAP activity, thereby enhancing overall repair capacity. To translate this finding into a therapeutic application, we developed a biohybrid heterospheroid (Mel-HS) by combining melatonin-loaded poly(lactic-co-glycolic acid) microspheres with 3D-cultured mesenchymal stem cells (MSCs), which serve as a PGE₂ source. We confirmed that this biohybrid construct preserves the paracrine capacity of MSCs to secrete PGE₂. Notably, Mel-HS demonstrates superior <i>in vivo</i> retention compared with naive 3D-MSCs, underscoring the cytoprotective effect of encapsulated melatonin in enhancing MSC viability. Furthermore, Mel-HS promoted robust RSC induction while simultaneously providing protection against inflammatory- and oxidative insults <i>in vitro</i>. In a colitis model, Mel-HS accelerated mucosal healing through the dual mechanisms-immunomodulation and enhanced RSC-driven repair-resulting in marked clinical improvement. Collectively, our findings highlight the therapeutic potential of enhancing endogenous regeneration with melatonin and MSCs, establishing a promising framework for next-generation biohybrid cell therapeutics in inflammatory bowel disease management.

The vomeronasal organ (VNO) is a specialized chemosensory structure that detects chemical cues involved in predator avoidance, social interactions and reproductive behaviors. In mice, the VNO contains distinct vomeronasal sensory neurons (VSNs) that express vomeronasal receptors (VRs) and formyl peptide receptors. Apical VSNs express Meis2, V1Rs and Gαi2, whereas basal VSNs express Tfap2e (also known as AP-2ε), V2Rs and Gαo. Type 2 VRs (V2Rs) are classified into families A-E, and basal neurons co-express a family C receptor with a VR from another family. Single-cell RNA-sequencing identified ∼980 genes differentially expressed between V1R- and V2R-expressing neurons, many of which are linked to endoplasmic reticulum (ER) functions. Notably, canopy 1 (Cnpy1) is highly enriched in V2R-expressing neurons. The VNO of Cnpy1 knockout mice develops normally but undergoes progressive loss of V2R-expressing neurons, which is associated with increased ER stress gene expression, upregulation of family C V2R mRNAs and reduced V2R protein levels. V2R-expressing VSNs in Cnpy1 knockouts fail to respond to pheromones, show altered guidance and adhesion gene expression, and display disrupted connectivity with the accessory olfactory bulb. These findings emphasize the role of cell-specific ER protein repertoires in maintaining neuronal function.

<h4>Background</h4>The litter size of goats is a key factor affecting their reproductive performance. The ovary plays a central role in the reproductive process of goats. However, the relationship between the dynamic changes in the ovarian proteome and the prolific traits of goats remains unclear. The Jining Grey Goat, an excellent dual-purpose meat and skin breed in China, is famous for its distinctive coat color characteristics and high reproductive performance.<h4>Methods</h4>It is an ideal animal model for studying goat reproductive performance. We selected 8 Jining Grey Goats and divided them into a single-birth group (n = 4) and a multiple-birth group (n = 4) to collect ovarian tissue samples. The iTRAQ technology, bioinformatics analysis, and Western blot were employed to investigate protein expression differences in ovarian tissues of goats with varying litter sizes.<h4>Results</h4>A total of 468 differentially expressed proteins (DEPs) were identified, of which 255 were up-regulated and 213 were down-regulated. GO enrichment analysis showed that these DEPs were mainly involved in biological processes such as cellular processes and metabolic processes, as well as molecular functions such as actin filament binding and calcium ion binding. KEGG pathway enrichment analysis revealed that the focal adhesion, Oxytocin Signaling Pathway, and PPAR Signaling Pathway were significantly enriched. Protein interaction network analysis identified CAV1, FLNA, CNN1, TAGLN, and MYH11 as key candidate proteins, and Western blot verification results were consistent with the iTRAQ trends.<h4>Conclusion</h4>This study revealed the molecular regulatory network of prolific traits in Jining Grey Goats from a proteomics perspective, providing a new insight into analyzing the genetic basis of goat fertility and serving as a reference for studying reproductive traits in other economically significant animals.

<h4>Background</h4>The concept of ferroptosis debuted as a newly defined programmed cell death in 2012. Among programmed cell death mechanisms, ferroptosis stands out as being fundamentally dependent on iron. At the heart of this mechanism lies the progressive accumulation of lipid peroxides - a chain reaction propelled by available iron, terminating when intracellular levels become fatally toxic. Inhibition of cystine transporters within cells (notably induced by compounds like Erastin) initiates a chain reaction: when intracellular levels of glutathione (GSH) become depleted, downstream suppression of <i>glutathione peroxidase 4 (GPX4)</i> activity impedes lipid peroxide clearance, whose accumulation drives the cell toward death upon exceeding a critical concentration.<h4>Discussion</h4>Early-stage experimental models highlight ferroptosis's contribution to propelling high-impact gynecological disease progression, namely precancerous endometrial hyperplasia, endometrial cancer (EC), endometriosis (EMS), and ovarian cancer (OC). Hence, elucidating the intricate regulatory machinery behind ferroptosis in gynecological pathologies bears both theoretical importance and translational promise.<h4>Conclusion</h4>This review aimed to systematically synthesize current knowledge on ferroptosis in gynecological diseases and their associated regulatory mechanisms, offering insights relevant to both basic research and clinical application.

<h4>Background</h4>Hepatic stellate cells (HSCs) play a pivotal role in driving fibrosis during chronic liver injury. HSCs produce vast amounts of fibrotic proteins, causing endoplasmic reticulum (ER) stress and initiating the unfolded protein response (UPR). While the UPR is important for fibrogenesis, how signaling through UPR transducer Protein Kinase R-like ER Kinase (PERK) and its effectors impact HSC activation and fibrogenesis is unclear. Here, we sought to uncover the role of PERK and its effector GADD45A in liver fibrosis.<h4>Methods</h4>PERK-GADD45A signaling was assessed in primary and immortalized HSCs treated with TGFβ, and mouse models of fibrosis. Genetic and pharmacological disruption of PERK or GADD45A was used to assess the role of PERK or GADD45A in HSC activation, proliferation, and fibrogenesis. HSC-specific Gadd45a-null mice were utilized to investigate the role of GADD45A on CCl4-induced fibrosis.<h4>Results</h4>We found that TGFβ-induction of collagen I drives activation of PERK signaling in HSCs. Furthermore, loss or inhibition of PERK limits long-term HSC activation as illustrated by reduced levels of collagen I and fibronectin, impaired collagen I deposition, and reduced cell proliferation in vitro. Next, we show that PERK signaling induces expression of GADD45A during HSC activation, and loss of GADD45A disrupts HSC activation and expression of proliferation and cell-cycle-associated genes in immortalized and primary HSCs. Finally, HSC-specific GADD4Aa loss limits CCl4-driven fibrogenesis in vivo.<h4>Conclusion</h4>PERK signaling is critical for HSC activation, and loss of the PERK downstream effector GADD45A limits fibrosis progression. Disruption of HSC activation and proliferation, coupled with dysregulation of cell-cycle-associated genes upon PERK or GADD45A loss, suggests that PERK-GADD45A signaling impacts multiple facets of HSC pathophysiology.

<h4>Background</h4>Electronic consultations (e-consults) improve specialist access but burden providers. We developed LiVersa, a customized large language model (LLM) for liver diseases. We evaluated its performance in drafting hepatology e-consult responses and the equivalence between human and machine reviewers.<h4>Methods</h4>LiVersa-generated responses for hepatology e-consults answered at the University of California San Francisco (UCSF) from January to March 2025. Using a 12-item rubric, 3 independent hepatologists and "LLM-as-a-judge" (OpenAI-o1) evaluated drafts against original responses. We tested equivalence between human reviewers and "LLM-as-a-judge" using two one-sided tests (TOST).<h4>Results</h4>Among 61 e-consults, the most common categories were abnormal liver function tests (34%), hepatitis B (23%), and abnormal imaging (21%). LiVersa drafts demonstrated no differences from hepatologist responses in word count (284 vs. 264, p=0.47) and verbosity (24 vs. 25 words per sentence, p=0.44). Human reviewers rated 72% of drafts as reasonable starting points and 83% as providing appropriate case-specific recommendations; 10% contained misleading/incorrect information, and 3.4% posed a risk of severe harm. LiVersa performed better at avoiding misleading information and extraneous suggestions but scored lower on clinical equivalence, immediate usability, and comprehensiveness. LLM-based reviewers were more stringent than human reviewers, rating fewer drafts as clinically equivalent (27% vs. 48%) and more as potentially harmful (67% vs. 20%), with agreement on accuracy, precision, and comprehensiveness (mean difference 0.026-0.029; TOST p<0.05).<h4>Conclusions</h4>Customized LLMs like LiVersa show promise for e-consult drafting but require human oversight. LLM-as-a-judge was more conservative than humans, supporting its role in rapid quality assurance during model updates.

<h4>Purpose</h4>Controlling bleeding is one of the most fundamental challenges in cardiac surgery. Bleeding is controlled by neutralising heparin with protamine. Our study investigates exosomes as an alternative treatment option to protamine for use in haemostasis.<h4>Methods</h4>Rats were divided into 5 groups, with 4 rats in each group: Group 1 underwent puncture, Group 2 underwent heparinised puncture, Group 3 received exosomes prior to heparinised puncture, Group 4 received exosomes prior to heparinised puncture with heparin neutralised by protamine, and Group 5 received heparinised puncture with exosomes administered during puncture. Biochemical analysis was made, histopathological analysis showing tissue damage and Mast Cell Activity (MAST )in lung tissues. Western blot method for SERPINC1 expression.<h4>Results</h4>Despite high Activated Clotting Time levels, bleeding time was found to be shorter in the group treated with exosomes. In MAST analyses, Group 4 had the highest density, while Group 3 had a density close to that of the control group. A significant difference was detected between Groups 3 and 4. in terms of lung tissue damage (P < 0.001). Tissue damage was most pronounced in Group 4. The application of exosomes significantly reduced inflammation and oedema in Group 3.<h4>Conclusion</h4>Exosomes shorten bleeding time and reduce lung tissue damage when administered before puncture. Exosomes have the potential to replace protamine in the future.

Autism spectrum disorder (ASD) is a heterogeneous neurodevelopmental condition. Studies of postmortem ASD brain tissue have revealed convergent molecular changes across the cortex. Whether these features are reflected in cell-type-specific epigenetic signatures is unknown. Here, we present a single-cell analysis of DNA methylation (DNAm) coupled with transcriptomics in ASD. Using snmCT-seq, we profiled DNAm and transcript levels from over 60,000 nuclei derived from the prefrontal cortex of 49 donors. We identified over 30,000 differentially methylated regions (DMRs) in ASD that were enriched in promoters and cell-type-specific regulatory elements active across the lifespan. ASD-related methylation changes were uncorrelated with transcript levels and were small in magnitude compared with age-associated effects. Age-DMRs were concentrated in excitatory neurons and revealed distinct roles for CG and non-CG methylation. Age-varying methylation signatures of ASD identified neuron projection development as a key process perturbed in ASD, highlighting the heterogeneous impact of ASD across the lifespan.

KRAS mutation is one of the most prevalent oncogenic driver mutations in NSCLC. UNC5C, as a member of dependence receptors of netrin-1, is a conditional tumor suppressor in cancer progression and metastasis via inducing apoptosis. Despite UNC5C has been intensively reported to be downregulated in multiple types of malignancies, the mechanism of UNC5C loss or downregulation in cancer remains unclear. In this study, we identified UNC5C as a downstream effector of oncogenic KRAS signaling pathway. We found that oncogenic KRAS suppressed UNC5C and inhibition of oncogenic KRAS upregulated UNC5C in KRAS-mutant NSCLC. Mechanically, oncogenic KRAS-mediated downregulation of UNC5C was dependent on the activation of the RAF/MEK/ERK cascade rather than the PI3K/AKT/mTOR pathway. More specifically, ERK2, but not ERK1, was involved in the control of UNC5C expression. Critically, FOS, a downstream transcription factor of the ERK pathway, was responsible for the transcriptional repression of UNC5C in KRAS-mutant NSCLC. In addition, UNC5C, rather than other dependence receptors of netrin-1, was most strongly downregulated in NSCLC and functioned as a tumor suppressor. In conclusion, we reported oncogenic KRAS-mediated transcriptional suppression of UNC5C and deciphered the exact underlying molecular mechanism in NSCLC, thus providing novel insights into the interplay between oncogenes and tumor suppressor genes in KRAS-driven NSCLC.

<h4>Background</h4>Immune-related adverse events (irAEs) induced by immune checkpoint inhibitors (ICIs) share symptomatic and therapeutic similarities with chronic inflammatory diseases (CIDs). However, the molecular distinctions between irAEs and CIDs remain unclear. Herein, we systematically compared irAEs and CIDs across multiple tissues using large-scale multi-omics profiling.<h4>Methods</h4>We compiled a transcriptomic compendium of over 4.3 million cells from 1137 samples and 24 spatial transcriptomes across diverse inflammatory sites in irAEs and CIDs. In addition, we collected 526 pre-ICI and post-ICI treatment-matched blood transcriptomes coupled with germline exomes, alongside 75 serum proteomes with irAE information.<h4>Results</h4>Hallmark gene signatures and cell states that characterize the inflammatory milieu of various irAEs and CIDs were defined. Across tissues, irAEs involve a distinct inflammatory ecosystem enriched with myeloid-derived components, including <i>NLRP3</i> ⁺ mononuclear phagocytes, <i>LAMP3</i> ⁺ dendritic cells, and <i>CXCL8</i>/<i>10</i> cytokine signaling, alongside <i>TNFRSF18</i> ⁺ regulatory T cell, T/natural killer interferon, and <i>HAVCR2</i> ⁺ exhausted T cell. These cell states exhibit unique interaction patterns, forming spatially localized niches specific to irAEs but absent in CIDs. Severe irAEs are accompanied by elevated <i>CXCL10</i> cytokine signaling post-ICI treatment, with a higher prevalence among <i>HLA-B</i>*<i>46:01</i> carriers.<h4>Conclusions</h4>Our multimodal analyses highlight key differences between irAEs and CIDs, indicating that irAEs constitute a distinct inflammatory ecosystem that differentiates them from CIDs. Our study provides insights into potential biomarkers and therapeutic targets for improved irAE management.

The APOEε4 allele and oestrogen deficiency independently predispose females to an increased risk of vascular and metabolic impairments, but their cerebrovascular effects are less understood. The purpose of this study was to determine the interaction between APOE genotype and oestrogen on cerebrovascular endothelial and mitochondrial function. We studied young female homozygous APOEε3 and APOEε4 mice (n = 19-20/group; ∼6 months old) that were fed a high-fat diet and were ovariectomized (OVX), OVX and supplemented with 17β-oestradiol, or left intact. In APOEε3 mice, OVX was associated with impaired posterior cerebral artery endothelium-dependent dilatation, which was rescued by 17β-oestradiol. However, in APOEε4 mice, there was no effect of OVX or 17β-oestradiol on cerebral artery endothelial function. Carotid artery passive stiffness was greater with OVX and lower with 17β-oestradiol treatment in APOEε3 mice, but there was no impact of OVX or 17β-oestradiol in the APOEε4 mice. In cerebral arteries and arterioles, 17β-oestradiol led to higher mitochondrial complex I respiration in APOEε3 but not APOEε4 mice. These functional differences were concomitant with group differences in mitochondrial DNA copy number, antioxidant enzymes and pro-inflammatory factors. Overall these results indicate that the APOE genotype modulates the impact of OVX and oestradiol on the cerebral vasculature. We found that 17β-oestradiol enhances cerebrovascular endothelial and mitochondrial function in OVX APOEε3 mice but not in APOEε4 mice. This suggests that 17β-oestradiol supplementation may have more cerebrovascular benefits for APOEε4 non-carriers. KEY POINTS: Females have twice the risk of Alzheimer's disease than males, and the APOEε4 genetic variant has a greater risk for Alzheimer's disease than the APOEε3 variant. The risk for Alzheimer's disease increases after menopause in females, suggesting that the loss of female sex hormones may play a role. There are highly inconsistent results among past studies examining the interaction between APOE genotype and oestrogens on brain outcomes, and their impact on the vasculature has not been studied. We aimed to determine the impact of APOEε4 genotype on the cerebrovascular response to ovariectomy and oestradiol. We found that oestradiol improved cerebral artery endothelial function and mitochondrial respiration in ovariectomized APOEε3 mice following ovariectomy. In contrast APOEε4 mice were resistant to the beneficial effects of ovarian hormones on cerebrovascular and mitochondrial function. This research suggests that APOE genotype may be a consideration when weighing the risks and benefits of prescribing hormone replacement therapy to postmenopausal females.

<h4>Background</h4>Chorea is a hyperkinetic movement disorder with a broad differential diagnosis, ranging from acute symptomatic causes to slowly progressive neurogenetic diseases. While Huntington's disease (HD) remains the most prevalent hereditary form, numerous other genetic disorders may mimic its clinical presentation. A major diagnostic challenge arises in patients with a seemingly negative family history, which can obscure the suspicion of a genetic etiology. In patients with sporadic chorea, the potential contribution of genetic testing to the diagnostic process has not yet been systematically analyzed.<h4>Methods</h4>We conducted a retrospective analysis of 81 patients presenting with chorea as a prominent symptom at the movement disorders outpatient clinic between 2013 and 2024. Clinical data, family history, laboratory results, imaging, and genetic analyses were evaluated. Genetic testing included a chorea-related gene panel and, if unremarkable, whole-exome or whole-genome sequencing.<h4>Results</h4>Out of 81 patients, 44 presented with slowly progressive chorea and unremarkable family history of HD or chorea-related syndromes. After exclusion of secondary etiologies (n = 8), 36 patients remained, of whom 30 (83, 33%) received a confirmed genetic diagnosis. HD was the most frequent diagnosis (n = 20), followed by rare genetic disorders such as Spinocerebellar Ataxia Type 17 (n = 2), Wilson's Disease (n = 2), Ataxia with Oculomotor Apraxia Type 2 (n = 1), C9orf72-related Neurodegeneration (n = 1), Choreoacanthocytosis (n = 1), KMT2B-related Dystonia (n = 1), ERCC4-related Neurodegeneration (n = 1), and Glutaric Acidemia Type 1 (n = 1).<h4>Discussion</h4>These findings support the systematic use of genetic testing-even in apparently sporadic cases-and suggest that the prevalence of hereditary choreatic disorders, may be significantly underestimated.

Muscle fibrosis is a major driver of progression in diverse myopathies, yet the conserved molecular mediators of this process in humans remain poorly defined. Here, we identify collagen VI as a common regeneration-impairing extracellular matrix (ECM) component across three distinct human myopathies: Duchenne Muscular Dystrophy (DMD), Oculopharyngeal Muscular Dystrophy (OPMD), and Inclusion Body Myositis (IBM). Proteomic profiling of fibrotic biopsies reveals consistent upregulation of collagen VI and laminin γ1, alongside disease-specific alterations. Fibroadipogenic progenitors (FAPs) are the predominant source of these ECM components, including collagen VI and laminin γ1. Functionally, xenotransplantation of patient-derived FAPs into regenerating mouse muscle induces localized collagen deposition, myofiber atrophy, and depletion of Pax7⁺ muscle stem cells. Mechanistic assays demonstrate that FAP-derived collagen VI is sufficient to impair myogenic fusion, while silencing COL6 in patient FAPs restores fusion capacity, directly linking pathological collagen VI deposition to regeneration failure. Our findings uncover collagen VI as a conserved effector of fibrosis and stem cell niche disruption in human myopathies, positioning it as a potential therapeutic target across genetically and clinically distinct muscle diseases.

<h4>Background</h4>The heterogeneity of lung adenocarcinoma (LUAD) makes it challenging to fully comprehend its development. Basement Membrane (BM) promotes cell polarity and has a function in cell development.<h4>Methods</h4>A single-cell transcriptional profile of LUAD was established to investigate the heterogeneity of the tumour microenvironment(TME). Subsequently, the cell differentiation status and intercellular communication analysis in LUAD microenvironment were analysed. A total of 250 genes were associated with the BM and involved in cell trajectory differentiation. Next, the variation in immune checkpoint gene expression levels and immunotherapy response between groups was analysed using consensus clustering. Weighted gene co-expression network analysis (WGCNA) was applied to identify genes from the modules that showed a strong correlation with clinical characteristics. Moreover, based on our thorough bioinformatics study, we established the prognostic signature of Basement Membrane -related genes (BMRGs) using genes with strong prognostic importance.<h4>Results</h4>The MIF signalling pathway was found to be significantly activated in the TME. Cell differentiation trajectory analysis identified five subpopulations with distinct differentiation states. Every cluster has distinctive individual clinical characteristics and reflects a different immunological microenvironment. A prognosis model was created using six BMRGs with the most significant predictive power for survival.<h4>Conclusion</h4>The model of risk score acts as a stand-alone prognostic factor, and the model is able to forecast patient prognoses with accuracy, which helps investigate more effective immunotherapy strategies.It also has a significant impact in TME.

<h4>Background</h4>Major depressive disorder (MDD) has been associated with large-scale functional dysconnectivity, yet it remains unclear whether these abnormalities differ as a function of anatomical distance. We investigated distance-related functional connectivity strength (FCS) alterations in first-episode, drug-naïve MDD and examined whether their spatial distribution corresponds to atlas-based neurotransmitter receptors/transporters profiles.<h4>Methods</h4>Resting-state functional MRI data were acquired from 191 patients with first-episode, drug-naïve MDD and 130 healthy controls. Whole-brain global, long-range, and short-range FCS were quantified voxel-wise. Distance-constrained seed-based connectivity analyses were performed to characterize the circuit-level contributions underlying abnormal FCS findings. To provide a molecular context, regional maps of FCS abnormalities were compared with neurotransmitter receptors/transporters distributions derived from a publicly available positron emission tomography atlas.<h4>Results</h4>Compared with healthy controls, patients with MDD showed reduced long-range FCS in the left orbital superior frontal gyrus and left medial superior frontal gyrus, together with increased short-range FCS in the right inferior parietal lobule. Seed-based analyses indicated that reduced long-range FCS was mainly related to weaker long-range connectivity between the left orbital superior frontal gyrus and right inferior temporal gyrus, and between the left medial superior frontal gyrus and right hippocampus. Increased short-range FCS in the right inferior parietal lobule was associated with stronger short-range connectivity with the right inferior frontal operculum, right superior temporal gyrus, and left paracentral lobule. The spatial pattern of FCS abnormalities showed significant correspondence with atlas-based neurotransmitter receptors/transporters distributions; short-range abnormalities spatially covaried with an atlas-derived regional excitation-inhibition balance.<h4>Conclusions</h4>First-episode, drug-naïve MDD is characterized by a distance-related pattern of reduced long-range prefrontal connectivity and increased short-range parietal connectivity. Atlas-based molecular mapping provides a biologically informed, population-level spatial context for these macroscale functional abnormalities.

<h4>Background</h4>Early diagnosis and targeted treatment of inner blood-retinal barrier (iBRB) impairment in non-proliferative diabetic retinopathy (NPDR) present significant challenges. This study investigates the cellular heterogeneity and early lesions in the iBRB microenvironment.<h4>Methods</h4>We created a single-cell transcriptional atlas of NPDR using retinas from Zucker Diabetic Fatty rats, focusing on the expression of cells within the iBRB microenvironment, particularly microglia. We performed cell-cell gene interaction analyses to investigate intercellular communications among different cell types in the iBRB. Additionally, we conducted differentiation potential and trajectory analysis, transcription factor regulatory network characterization, and enrichment analysis of microglia. We also employed transmission electron microscopy, immunofluorescence, and histological analysis to validate the hypothesis.<h4>Results</h4>Based on retinal samples from NPDR rats which had retinal edema and kidney damage, and normal rats, we selected 36,821 cells for subsequent analysis using single-cell sequencing. We identified 669 cells in iBRB and microglia had the highest connectivity value by enrichment analysis. 980 differentially expressed genes related to microglia divide it into 4 subtypes. Among them, Spp1 highly expressed microglia subtype 2 had the highest differentiation potential and was significantly upregulated after NPDR lesions. By integrating histological and biochemical analysis results, the roles of the subtype in the differentiation of microglia in the iBRB microenvironment, such as oxidative stress and apoptosis, were well validated.<h4>Conclusion</h4>This study identified a novel microglia type in NPDR rats, offering valuable insight into targeted therapeutic strategies for NPDR patients and enhancing our understanding of microglial regulation within the iBRB.

<h4>Background</h4>Mitochondrial proteases are essential for mitochondrial protein import and constitute the core of the organelle's intrinsic protein quality control system. However, their physiological functions across tissues, as well as their influence on cytosolic proteostasis, remain incompletely understood.<h4>Results</h4>We generated loss- and gain-of-function alleles for 15 conserved mitochondrial proteases in Drosophila melanogaster to systematically dissect their in vivo functions. Disruption of specific proteases caused male sterility or organismal lethality, whereas tissue-specific knockouts in the eye, muscle, or fat body led to mitochondrial protein aggregates, structural defects, and age-dependent degeneration. Loss of UQCR-C1 or Afg3l2 robustly increased mitophagy, while overexpression of several proteases severely impaired muscle integrity. Loss of UQCR-C1, Mppa, or CG11771 promoted HTT72Q aggregation, and reducing UQCR-C1 or Afg3l2 markedly elevated cytosolic HTT72Q levels. Conversely, overexpressing Mppa-but with reduced efficacy in its disease-associated variants-suppressed HTT96Q aggregation and neuronal toxicity. Mppa forms a complex with UQCR-C1 to regulate mitochondrial pre-protein processing and import, indicating that enhancing mitochondrial protein import is sufficient to alleviate cytosolic proteotoxic stress caused by HTT polyglutamine (polyQ) proteins.<h4>Conclusions</h4>This work establishes a comprehensive in vivo resource for mitochondrial protease functions and their roles in shaping cytosolic proteostasis.

Smart DNA hydrogels have attracted extensive attention in bioanalysis and biomedicine due to their programmable sol-gel phase transition ability and specific recognition characteristics of biomolecules. Herein, we propose a spatially confined "double-key lock" smart DNA hydrogel with temperature and target as double keys, which was constructed using poly(<i>N</i>-isopropylacrylamide)-acrylamide (NIPAM-AM) hydrogel as the carrier and aggregation-induced emission (AIE) type 1,1,2,2-tetra(4-carboxybenzene) ethylene (TCPE) as the fluorescence (FL) probe. This hydrogel exhibited a synergistic multi-response mechanism that combined the thermal phase transition of NIPAM-AM hydrogel, and the target recognition effect of complementary DNA strands. Upon concurrent stimulation by temperature elevation and target cDNA binding, this hydrogel underwent enhanced crosslinking, forming a denser three-dimensional network that restricted TCPE mobility and amplified its FL through aggregation. Molecular dynamics simulations further confirmed that the response mechanism involved the synergistic effect of thermodynamic equilibrium regulation and molecular configuration rearrangement. The combination of the smart DNA hydrogel and the rolling circle amplification (RCA) strategy realized the FL nucleic acid analysis of microRNA-21 (miRNA-21) as an analyte, exhibiting a wide linear response in the range of 0.1 fM to 10 nM with a limit of detection of 33.3 aM. Moreover, this platform was successfully applied to the quantitative analysis of miRNA-21 in the serum of lung cancer and gastric cancer patients, and can <i>in situ</i> track the secretion behavior of tumor cell derived miRNA-21. This strategy provides new insights into the multi-scale design of temperature-biomolecular double-responsive smart nanomaterials and offers a promising strategy for advanced biosensing and bioanalytical applications.

Rapid technological advancement has led to a growing need for new materials with enhanced properties to meet the demands of emerging applications. Traditional single-component materials often fail to meet the required demands, pushing the exploration of new composite materials. In these contexts, polyhedral oligomeric silsesquioxanes (POSS) as hybrid nanomaterials exhibit excellent mechanical, thermal and chemical stability, making them ideal for advanced material design. In addition, their hydrophobic nature and ability to be functionalized enable antimicrobial properties with enhanced resistance to microbial adhesion. Compared to hybrid nanomaterials, single-component antiviral materials have limitations, including short-term durability and low surface stability. The rationale for developing the POSS-stearic acid hybrid composite is based on the ability to form a rigid nanocage framework of POSS, which provides structural robustness and enhanced stability. We show that by combining POSS with stearic acid, the hybrid material displays greater hydrophobicity than its individual components, making it an effective surface coating material that prevents the attachment of microbial pathogens, including viruses, such as human coronavirus-OC43 (HCoV-OC43) and Type A influenza virus (H1N1). We provide evidence that the present material not only serves as a barrier to viral attachment to the surface but also significantly reduces viral infectivity, possibly through direct neutralization, thereby offering a promising strategy for mitigating surface-mediated transmission of respiratory viruses.

Per- and polyfluoroalkyl substances (PFAS) are persistent, chemically stable compounds widely used in daily life. Perfluorooctanoic acid (PFOA), perfluorononanoic acid (PFNA), perfluorohexanesulfonic acid (PFHxS), and perfluorooctanesulfonic acid (PFOS) were identified as the most relevant PFAS due to their prevalence and toxicity. This study aimed to investigate the immunotoxic mechanisms of a mixture of these PFAS using an in silico approach. Comparative Toxicogenomic Database (CTD), GeneMANIA, CytoHubba (Cytoscape), ToppGene Suite, and Metascape were used for the analysis. A total of 65 immune-related genes were identified as common to all four PFAS, with <i>IFNG</i>, <i>TNF</i>, <i>IL1B</i>, <i>IL6</i>, <i>TYK2</i>, <i>CD3E</i>, <i>CASP8</i>, <i>VAV1</i>, <i>ARHGAP4</i>, and <i>CARD11</i> emerging as key hub genes. CTD phenotype analysis indicated immune dysregulation, with decreased humoral and adaptive immune responses in humans and tissue-specific modulation of B- and T-cell activity in mice, while no immune-related phenotypes were observed for PFNA. Network analysis identified functional modules associated with apoptotic and immune signaling, endothelial cell migration and angiogenesis, and shared inflammatory and viral response pathways. Disease enrichment analysis associated PFAS with autoimmune disorders (rheumatoid arthritis, asthma), metabolic conditions, and cardiovascular diseases (experimental diabetes, hypertensive disease). These results highlight PFAS involvement in immune modulation, cytokine signaling, and disease susceptibility.

Short cytosine-adenine-guanine (CAG) trinucleotide repeats, which encode polyglutamine (polyQ) tracts, are prevalent features of genes enriched in transcriptional and regulatory functions, including the androgen receptor (AR) and huntingtin (HTT). While expanded CAG repeats are well established in neurodegenerative disease pathogenesis, the functional significance of short, non-pathogenic repeat lengths remains underappreciated. This review connects evidence demonstrating that short CAG/polyQ tracts act as dynamic modulators of protein conformation, transcriptional activity, and protein-protein interactions. Variation within physiological repeat ranges influences receptor sensitivity, cellular signaling, and phenotypic diversity. The AR serves as a central model, where shorter repeat lengths enhance transactivation and androgen responsiveness and are associated with increased prostate cancer risk, while longer non-expanded repeats are linked to reduced receptor activity and modest reproductive and metabolic effects. Mechanistically, repeat length and sequence composition jointly influence repeat stability, RNA structure, and downstream regulatory processes. Beyond AR, short CAG variation contributes to neuropsychiatric phenotypes and broader regulatory networks. Collectively, short CAG repeats function as quantitative regulators of gene activity, shaping disease susceptibility, physiological variation, and evolutionary adaptation.

Sarcoids are the most common cutaneous tumors in horses, representing up to 90% (35%-90%) of skin neoplasms. Mostly caused by Bovine Papillomavirus (BPVs) infections, sarcoids are highly resistant to therapy and prone to recurring, posing a significant threat to equine health. The aim of this study is to explore molecular pathogenetic mechanisms underlying the development of equine sarcoids, by applying transcriptomic approach. After testing samples for viral DNA, both mRNA and small RNA expression was analyzed via high-throughput Illumina sequencing comparing 12 sarcoids and 12 healthy skin samples as controls. Differentially expressed genes (DEGs), DE miRNAs (sarcoids vs. controls) and miRNA-DEG couples with opposite expression trends, were retrieved and subjected to a functional analysis. Over 6K DEGs emerged, 3620 down-regulated and 2415 up-regulated along with 145 DE miRNAs, 56 downregulated and 89 upregulated. Among the enriched biological processes for DEGs, some were related to growth factors production and collagen binding, cell migration and proliferation, tissue morphogenesis and inflammatory response. Interestingly, "Pathways in cancer" and "Hippo signaling pathway" were enriched KEGG pathways for the miRNA-DEG couples. Our data identified a great transcription discrepancy between sarcoid lesions and healthy skin with an overall enrichment for processes related to cellular transformation. RNA-seq sequencing depth allowed the search for candidate chimeric transcripts associated with viral integration events. Chimeric RNAs can influence gene regulation and may contribute to tumor growth and immune modulation. Via computational analysis we identified six fusion loci in tumor samples and in two sarcoid margins, with the most frequent event involving WNT10B and FKBP11. This fusion, detected in 6/10 sarcoids, is of particular interest since WNT10B activates the WNT/β-catenin cascade, while FKBP11 has been implicated in osteosarcoma progression. Although functional validation is ongoing, this represents the first report of chimeric transcripts in equine sarcoids, opening new perspectives on BPV-driven oncogenesis.

<h4>Background</h4>Major depressive disorder (MDD) is recognized as a pressing global public health burden. However, its molecular mechanisms remain incompletely understood.<h4>Methods</h4>In this study, an integrative analysis of transcriptome datasets from the GEO database was conducted. GEO2R and the R programming language were used to identify differentially expressed genes (DEGs) related to oxidative stress and mitophagy. Key hub genes, such as <i>EEF2</i>, <i>CCT3</i>, <i>EIF3I</i>, and <i>RPS5</i>, were further identified through enrichment analysis and protein-protein interaction (PPI) network construction. Following validation using an independent human dataset, we established a corticosterone-induced C8-D1A cell model. Reactive oxygen species and mitochondrial membrane potential were measured via flow cytometry. The results demonstrated that this model reliably recapitulates key pathological features of elevated oxidative stress and mitochondrial dysfunction in MDD. Finally, using an <i>in vivo</i> mouse model, we assessed synapse-associated proteins and mitophagy markers using Western blotting and measured the mRNA expression levels of candidate genes by qPCR to comprehensively validate the associations between the expression of the aforementioned genes and oxidative stress, mitophagy, and synaptic damage.<h4>Results</h4>This study combined bioinformatics screening and multidimensional phenotypic validation to construct an MDD-specific molecular regulatory network focused on carbon metabolism, thereby elucidating the interplay between four genes and oxidative stress and mitophagy. Although <i>CCT3</i> and <i>RPS5</i> demonstrated modest diagnostic utility in the independent validation dataset (AUC ≈ 0.6, <i>Padj</i> < 0.05), subsequent <i>in vivo</i> experiments revealed that the mRNA expression levels of these genes were significantly downregulated in MDD models (<i>EEF2</i>: <i>P</i> < 0.05; <i>CCT3</i>: <i>P</i> < 0.005; <i>EIF3I</i>: <i>P</i> < 0.05). Furthermore, the expression levels of these genes were positively correlated with those of synaptic proteins and negatively correlated with those of mitophagy markers. The downregulation of these genes may impair protein synthesis and folding, which acts in synergy with oxidative stress and mitochondrial dysfunction to perpetuate the vicious cycle of bioenergetic crisis and proteostasis collapse in MDD.<h4>Conclusion</h4>Although this study did not experimentally validate the regulatory functions of the target genes or identify highly specific diagnostic biomarkers, it offers a novel molecular perspective for deciphering the complex pathology of MDD. Notably, this highlights the synergistic interaction between translational regulation and metabolic homeostasis. Further validation in larger independent cohorts is warranted to assess the viability of these genes as mechanistic therapeutic targets.

Myeloid sarcoma (MS) is a rare extramedullary tumor characterized by the proliferation of immature myeloid precursor cells. It most commonly occurs in association with acute myeloid leukemia, either at initial diagnosis or during disease progression. Breast involvement is exceedingly rare and represents a significant diagnostic challenge, as it can closely mimic more common breast malignancies, particularly carcinoma or lymphoma, frequently leading to misdiagnosis. We report the case of a 39-year-old woman who presented with two bilateral breast masses, initially suspected to be malignant based on clinical and radiological findings. Histopathological examination revealed a poorly differentiated malignant neoplasm composed of immature cells with eosinophilic cytoplasm and prominent nucleoli, diffusely infiltrating the mammary parenchyma. Immunohistochemical analysis demonstrated strong positivity for leukocyte common antigen (LCA), myeloperoxidase (MPO), and CD68, with focal expression of CD34 and CD117, supporting the diagnosis of primary bilateral breast MS; notably, aberrant expression of PAX5, a marker usually associated with B-cell lineage, constituted a potential diagnostic pitfall. Molecular analysis further identified a KMT2A::MLLT10 fusion, a rearrangement associated with monocytic differentiation and extramedullary involvement. Bone marrow biopsy showed no evidence of neoplastic infiltration. The patient was treated with combined chemotherapy and radiotherapy. This case highlights the diagnostic challenges posed by bilateral breast MS and underscores the critical role of histopathology, immunohistochemistry, and molecular studies in establishing an accurate diagnosis and guiding appropriate management.

Biocatalysis has proved to be of great importance to pursue a more sustainable production of fine chemicals, and enzyme immobilization is a crucial tool to achieve this goal. In this work, hydroxyapatite, an eco-friendly material, was used as a support for the immobilization of a γ-glutamyltransferase from <i>Escherichia coli</i>, an enzyme that catalyzes the synthesis of bioactive γ-glutamyl derivatives, exploiting the noncovalent interactions between the support and the enzyme. After screening the immobilization conditions, the storage (up to 90 days) and the thermal stability (50 °C) of the obtained immobilized biocatalyst were studied, as well as its reuse in up to 10 consecutive reactions of γ-glutamylation of <i>S</i>-allyl-l-cysteine, to give γ-l-glutamyl-<i>S</i>-allyl-l-cysteine (<b>3</b>), a flavor enhancer occurring in garlic extract. Finally, the immobilized enzyme was used to prepare a packed-bed reactor and <b>3</b> was synthesized under continuous-flow conditions improving both the productivity (from 19.4 to 35.1 μmol·h^-1) and the space-time yield (9.7 vs 381 μmol·h^-1·mL^-1) of the reaction.

<h4>Background</h4>Radiation-induced enteropathy represents a key limitation to recovery after accidental radiation exposure or radiotherapy, with few effective pharmacological treatments. We investigated the therapeutic potential of ginsenoside Rb1, a major saponin from <i>Panax ginseng</i>, in attenuating the radiation-induced intestinal injury.<h4>Methods</h4>C57BL/6 mice were exposed to abdominal irradiation at a dose of 13.5 Gy and treated with Rb1. The epithelial integrity, bacterial translocation, and immune modulation were assessed. <i>In vitro</i>, rat intestinal epithelial cell line, IEC-6 were examined for proliferation and Wnt/β-catenin activation in response to Rb1 and IL-10. The role of IL-10 was validated using neutralization experiments.<h4>Results</h4>Rb1 significantly alleviated radiation-induced crypt damage, villus shortening, bacterial translocation, and inflammation. It enhanced regulatory T (Treg) cell population and IL-10 secretion, which in turn activated Wnt/β-catenin signaling and promoted epithelial regeneration. <i>In vitro</i>, IL-10, but not Rb1 directly, promoted the proliferation of irradiated IEC-6 cells, whereas IL-10 neutralization abolished the mitigative effect of Rb1 in vivo.<h4>Conclusion</h4>Rb1 mitigated radiation-induced enteropathy by upregulating Treg abundance and IL-10 signaling, which in turn activated epithelial regeneration with Wnt/β-catenin pathway activation, supporting its potential as a therapeutic agent for radiation-induced intestinal injury.

Huntington’s disease (HD) is an autosomal dominant neurodegenerative disorder caused by a CAG-repeat expansion in the HTT gene. Progressive loss of striatal projection neurons leads to cognitive, psychiatric, and motor impairments that typically manifest in midlife, despite the presence of the expansion from conception. Increasing evidence supports a neurodevelopmental component to HD; however, authentic human developing HD striatal tissue has not previously been characterised. We analysed an HD positive human fetal striatal sample alongside an age- and sex-matched control. CAG-repeat length was determined, and single-cell RNA sequencing was used to investigate gene expression. We compared the fetal HD transcriptional signature with publicly available datasets from postmortem adult HD brain tissue. We identified 2,032 differentially expressed genes and defined nine cellular clusters, each exhibiting distinct transcriptional profiles. Gene enrichment analysis revealed disruption of key biological processes across the developing HD striatum, with pathway-level dysregulation varying between clusters. There was overlap in gene expression changes between fetal and adult HD striatal tissues. Together, these findings demonstrate that molecular features of HD pathology are present during early human striatal development, supporting the concept that disease mechanisms are established decades prior to clinical onset.

Antibiotic-associated diarrhea (AAD) remains a common complication of antibiotic therapy. While probiotics show therapeutic potential, the novel strain Bacillus subtilis THC1I has not been previously evaluated for AAD treatment. This study aimed to assess the effects of B. subtilis THC1I on intestinal barrier integrity and gut microbiota dysbiosis in an AAD mouse model. Fifty mice were randomized into five groups (n = 10): control, model, positive control (Enterogermina®, B. clausii), and a B. subtilis THC1I spore suspension at 0.82 × 10⁹ and 1.64 × 10⁹ CFU/kg/day. AAD was induced with gentamycin sulfate and cefradine for five consecutive days. Outcomes included clinical symptoms, hematological and biochemical parameters, colonic macroscopic and histopathological indices, inflammatory cytokines, and gut microbiota analyzed by 16S rRNA sequencing. B. subtilis THC1I significantly improved body weight, water intake, fecal scores, and fecal water content. Treatment restored electrolyte balance (sodium and potassium), reduced white blood cell counts, and decreased relative colon weight and inflammation scores. Histopathological analysis revealed restored epithelial architecture and increased goblet cell density. Pro-inflammatory cytokines (TNF-α, IL-1β) were significantly reduced. B. subtilis THC1I partially improved microbial diversity (Shannon index) and modulated microbiota composition at both phylum and genus levels, decreasing pathogenic bacteria (Proteobacteria phylum: Escherichia-Shigella, Klebsiella, Salmonella; Firmicutes phylum: Clostridioides), and modulating the dysbiotic overgrowth of the beneficial commensal Blautia, while promoting beneficial bacteria (Bacteroidota phylum: Bacteroides, Muribaculaceae; Firmicutes phylum: Lachnospiraceae, Lactobacillus). B. subtilis THC1I demonstrates restorative effects on intestinal barrier damage and gut microbiota dysbiosis in AAD mice, supporting its potential as a therapeutic candidate for clinical application.

Human γ9δ2 T cells detect microbial phosphoantigens (pAg) through butyrophilin (BTN) family proteins; however, how multiple BTN paralogs assemble into functional signaling complexes remains unresolved. Because γ9δ2 T cell activation requires coordinated action of several BTN members rather than a single receptor pair, defining their organization in living cells is essential. Here, we apply a live-cell NanoBRET saturation analysis using a novel HaloTag ligand to quantitatively resolve BTN oligomerization dynamics. We found that BTN3A1 exists as higher-order basal oligomers. Coexpression of BTN3A2 or BTN3A3 redistributes these assemblies into defined BTN3 dimers. The BTN3 dimers have a baseline interaction with BTN2A1 dimers that is altered by IgV domain interactions and by classical pAgs such as (<i>E</i>)-4-hydroxy-3-methyl-but-2-enyl diphosphate. Mutation of a BTN3A1 IgV epitope disrupts a restraint on BTN2A1 and triggers spontaneous γ9δ2 T cell activation, which is independent of pAg. In contrast, BTN3A3 forms a strong, constitutive tetramer with BTN2A1 in the absence of BTN3A1/BTN3A2. This complex potentiates detection of nonclassical pAgs such as mevalonate diphosphate, and disruption of its juxtamembrane/B30.2 intersection abolishes both complex formation and signaling. BTN3A2 suppresses both interaction and function of BTN3A3 complexes, in contrast to its stimulatory effect on BTN3A1 complexes. Together, these results reveal that γ9δ2 T cell activation is controlled by ligand-dependent activation of competing BTN complexes. More broadly, this work establishes quantitative NanoBRET analysis as a general framework for dissecting multiprotein membrane receptor organization in living cells.

GPR3, GPR6, and GPR12 form a subfamily of Class A orphan G protein-coupled receptors (oGPCRs), for which endogenous ligands have not been identified. Despite their high sequence similarity, each receptor exhibits unique expression profiles in human tissues. Their physiological roles and therapeutic potential are gradually being understood, indicating their critical involvement in various diseases, including central nervous system (CNS) disorders, metabolic diseases, and cancer. Notably, the GPR6 inverse agonist CVN424 is currently in Phase III clinical trials for the treatment of Parkinson's disease (PD). The recent determination of high-resolution structures of GPR3, GPR6, and GPR12 has significantly enhanced their attractiveness as emerging therapeutic targets for drug discovery. Herein, we summarize the current understanding of the structural and functional characteristics of GPR3, GPR6, and GPR12. We further highlight recent progress in relevant ligand discovery and discuss the key challenges and opportunities in developing potent and selective modulators targeting these orphan receptors.

Polycomb Repressive Complex 2 (PRC2), containing homologous EZH1 or EZH2 as the catalytic subunit, is a conserved methyltransferase complex that represses gene transcription by transferring methyl group from SAM to H3K27. Gain-of-function mutations of EZH2 and aberrant H3K27 methylation have been linked to human cancers. SAM-competitive EZH1 and EZH2 dual inhibitors have been approved by the FDA for treating B-cell lymphoma and other cancers. Here, we characterized a small molecule C36, which potently inhibits EZH2/PRC2, but not EZH1/PRC2, with a novel SAM non-competitive mechanism. Cryo-EM structures revealed that C36 binds to a pocket at the interface of SET-Activation-Loop (SAL), stimulation-responsive motif (SRM), and I-SET domain of EZH2, and WD40 domain of EED. C36 binding induces conformational changes and disrupts allosteric communication between EZH2 and ligand-bound EED. C36 efficiently inhibits H3K27 trimethylation and PRC2 target gene expression in tumor cells and xenograft tumors with low hematotoxicity. Multi-omics analyses employing C36 uncovered the direct regulation of IFNB1 by EZH2/PRC2. The combined treatment of syngeneic LLC lung cancer with C36 and a PD-1 antibody significantly enhances anti-tumor efficacy. Our study identifies a new allosteric mechanism of PRC2 inhibition and paves the way for the development of highly selective EZH2/PRC2 inhibitors for combination therapy.

Japanese encephalitis virus (JEV) is a neurotropic flavivirus that causes a substantial threat to human health and livestock; however, the epitranscriptomic mechanisms that support its replication remain poorly defined. Here, we identify a proviral host factor C₂H₂ zinc-finger protein ZNF33B that promotes JEV infection through coupling N⁶-methyladenosine (m⁶A) RNA modification to autophagy regulation. Mechanistically, ZNF33B recruits METTL14 to stabilize the METTL3-METTL14 methyltransferase complex, thereby increasing global m⁶A deposition. Multi-omics analyses reveal that ZNF33B selectively binds m⁶A-modified sites within the antiviral transcript Trim25 (c.1567 and c.1669 bp) to accelerate its decay. We further demonstrate that TRIM25 functions as an E3 ubiquitin ligase that catalyzes K48-linked ubiquitination of ATG7 at lysines 389 and 423, leading to its proteasomal degradation and ultimately suppressing autophagic flux. In contrast, ZNF33B-mediated Trim25 degradation counteracts its inhibitory effect on autophagy, creating a favorable environment for viral replication. In vivo, adeno-associated virus (AAV)-mediated ZNF33B delivery increases mouse brain m⁶A levels, decreases TRIM25 expression, elevates ATG7 abundance, exacerbates JEV-induced neuropathology, and accelerates mouse mortality. Together, these findings reveal a previously uncharacterized ZNF33B-m⁶A-TRIM25-autophagy axis that JEV hijacks to evade host antiviral responses, providing new insights into flaviviral pathogenesis and potential therapeutic targets.

Hyperuricemia (HUA) affects diverse biological processes and signaling pathways across multiple organ systems; however, its impact on the intestine remains poorly understood. Here, we show that HUA disrupts intestinal barrier function primarily by impairing intestinal stem cell (ISC) function, which is essential for epithelial renewal. Mechanistically, elevated uric acid (UA) directly binds to the mitochondrial protein CCDC90B, leading to excessive mitochondrial ROS accumulation, activation of the NLRP3 inflammasome, and subsequent initiation of downstream pyroptotic signaling. The resulting exhaustion of the intestinal stem cell pool impairs epithelial regeneration and further weakens intestinal barrier integrity. Collectively, these findings reveal a previously unrecognized mechanism linking UA to ISC dysfunction and highlight CCDC90B as a potential therapeutic target for HUA-associated intestinal dysfunction.

During the initial workup of a patient with chronic heart failure, increased left ventricular filling pressures at rest or during exercise, and a left ventricular ejection fraction ≥50%, physicians are expected to exclude the presence of established diseases that masquerade as heart failure with preserved ejection fraction (HFpEF), referred to as "HFpEF mimics." These diseases include (1) cardiac amyloidosis; (2) hypertrophic cardiomyopathy; (3) infiltrative, restrictive, and inflammatory cardiomyopathies (sarcoidosis, hemochromatosis, Fabry disease, and radiation- and chemotherapy-induced restrictive disorders); (4) hemodynamically significant valvular heart disease; (5) pericardial diseases (particularly constrictive pericarditis); (6) high-output heart failure; and (7) end-stage kidney disease. These diseases were recognized long before HFpEF was defined as a disorder, have specific targeted treatments, and have generally been excluded from randomized controlled trials of new drugs for HFpEF. Interestingly, after the exclusion of HFpEF mimics, patients demonstrate the typical clinical and pathophysiologic features of cardiometabolic HFpEF, with an exceptionally high prevalence of central adiposity, dysglycemia, hypertension, and systemic inflammation-even though large-scale trials did not require participants to have any feature of a cardiometabolic disorder. The homogeneity of the phenotypic presentations of these patients is consistent with the lack of observed heterogeneity in the responses to broad-based treatments (ie, sodium-glucose cotransporter 2 inhibitors, finerenone and incretins). Importantly, many patients with adiposity-related HFpEF were not permitted to participate in large-scale trials, which excluded patients with severe obesity, lower natriuretic peptides and without manifest echocardiographic diastolic filling abnormalities. In conclusion, the exclusion of HFpEF mimics from the broad population of patients with left-sided heart failure and an ejection fraction ≥50% yields a relatively homogeneous patient population with the features of adiposity-related HFpEF.

<h4>Background</h4>Melanoma shows one of the highest response rates to immune checkpoint inhibitors (ICIs), yet nearly half of patients experience primary or acquired resistance. While immune contexture strongly influences therapeutic efficacy, tumor cell-intrinsic features are increasingly recognized as key regulators of antitumor immunity. In particular, intratumoral heterogeneity driven by melanoma cell plasticity underlies diverse immune escape mechanisms. How this plasticity shapes ICI outcomes in patients remains poorly defined.<h4>Methods</h4>Tumor cell states and immune contexture were assessed in 57 primary cutaneous melanomas from stage III patients, collected prior to adjuvant anti-programmed cell death protein-1 (anti-PD-1) therapy and stratified according to 2-year relapse status (33 relapse-free, 24 relapsed). Whole slide multiplex immunofluorescence was combined with spatial transcriptomics (Visium, n=4) to investigate the spatial architecture of melanoma cell states, T cells, tumor-associated macrophages (TAMs), dendritic cell subsets, and tertiary lymphoid structures.<h4>Results</h4>Unsupervised clustering of melanoma cells identified distinct phenotypic states that formed spatially restricted homotypic patches. From these data, we defined a melanoma plasticity ratio (undifferentiated/differentiated tumor patches), which was significantly associated with reduced relapse-free survival. Integrated immune analyses recapitulated prognostically distinct immunotypes, with macrophage subsets displaying striking spatial compartmentalization. Antitumoral macrophages preferentially infiltrated differentiated melanoma regions, while protumoral macrophages localized to undifferentiated patches. Spatial transcriptomics confirmed that melanoma cell states tightly shape the neighboring immune microenvironment, with macrophages emerging as pivotal players. Their polarization was further influenced by tumor-derived signals in addition to microenvironmental cues (interferon-gamma, hypoxia). Entropy-based integration of melanoma cell states, TAMs, and T cell subsets uncovered two dominant spatial ecosystems with opposing associations to ICI efficacy. Ecosystems enriched in differentiated melanoma cells, programmed death-ligand 1 (PD-L1)⁺ TAMs, and PD-1⁺CD8⁺, and CD4⁺ T cells correlated with favorable outcomes, whereas ecosystems composed of undifferentiated melanoma cells with PD-L1^- protumoral TAMs and PD-1^-CD8⁺ T cells were associated with relapse.<h4>Conclusions</h4>Our study uncovers how cancer cell plasticity shapes spatially organized tumor-immune ecosystems that critically modulate adjuvant ICI efficacy in melanoma. These findings highlight melanoma cell plasticity as a key driver of immune evasion via macrophages reprogramming, through targetable interactions that may represent novel therapeutic avenues to enhance ICI efficacy.

Adaptation to chronic hypoxia (CH) enhances myocardial tolerance to ischemia/reperfusion injury and is closely associated with stabilization of hypoxia-inducible factor-1 alpha (HIF-1α), a central transcription factor in hypoxic response. Given the central role of mitochondria in cardiac pathophysiology, we investigated the contribution of HIF-1α to cellular mechanisms underlying CH-induced cardioprotection, with a focus on proteomic remodeling, antioxidant defense, and regulation of the mitochondrial permeability transition pore (mPTP). Adult male wild-type and heterozygous Hif1a knockout mice were exposed to intermittent CH (7000 m, 8 h/day, 4 weeks) or kept under normoxia. Isolated perfused hearts treated with cyclosporine A, an inhibitor of mPTP opening, were subjected to global ischemia/reperfusion insult for infarct size determination. Quantitative label-free proteomics was conducted to assess HIF-1α-dependent changes. We evaluated oxidative stress, measured levels of proteins associated with antioxidant defense and mPTP regulation. In parallel, a proof-of-concept study was performed in transfected AC16 cardiomyocytes exposed to H₂O₂-induced oxidative stress. CH induced HIF-1α-dependent cardioprotection by limiting infarct size through regulation of mPTP opening. Moreover, CH attenuated oxidative stress and promoted the protective translocation of hexokinase-2 to mitochondria in an HIF-1α-dependent manner. Consistently, HIF-1α overexpression enhanced cardiomyocyte survival under oxidative stress, whereas HIF-1α inhibition by acriflavine reduced cell viability. These findings identify HIF-1α as a key mediator of CH-induced adaptive response and cardioprotection during I/R injury in the mouse heart, acting through mPTP regulation.

The current study establishes the mechanism on how dietary flavonoid, catechin fractions of Osbeckia parvifolia Arn. could be used as a therapeutic treatment for ovarian cancer, using a combination of network pharmacology, molecular validation and nano-formulations. From network modelling analysis, 420 protein targets linked to ovarian cancer, including five that act as major regulatory hubs - AKT1, STAT3, IL6, ESR1 and CASP3 were found. Catechin had the strongest binding affinity to all of these five hub proteins, especially for ESR1 (-11.089 kcal/mol) and AKT1 (-9.221 kcal/mol). In molecular dynamics simulation studies over 100 ns, catechin bonded strongly with its target proteins. HPLC guided fractionation (RT: 3.887 min) LC-MS/MS (RT:6.51 min), HPTLC (Rf: 0.973) were performed, which resulted in the confirmation of the presence of both catechin and epicatechin and verified the biosynthetic pathway of both through the measurement of the expression of their respective biosynthetic genes LAR (0.3637 folds) and ANR (0.2746 folds) in O. parvifolia. The evaluation of the six different nano-formulations of phytoniosomes led to F5 being the most stable with uniform spherical architecture, having an optimal particle size (229.2 nm), a high positive zeta potential (+ 42.7mV), and having the highest cytotoxicity (93.17%) in SKOV3 cells, the mechanism of action being through apoptosis induction and cell-cycle arrest. Hence, this study supports the hypothesis that catechin extracted from O. parvifolia can serve as an effective therapeutic candidate for the targeted treatment of ovarian cancer.

<h4>Background</h4>Long-term proton pump inhibitor (PPI) use is associated with increased intestinal disease risk, but its damaging mechanisms remain unclear.<h4>Methods</h4>Mice were administered rabeprazole (Rab) for 4 weeks before dextran sulfate sodium (DSS) or ionizing radiation (IR) injury. We employed RNA sequencing, metabolomics, and metagenomics, evaluated intestinal stem cell (ISC) function, and used organoids for validation.<h4>Results</h4>Long-term Rab induced small intestinal mucosal injury and exacerbated DSS/IR-induced damage, manifesting as crypt/villus atrophy and reduced ISC numbers. Mechanistically, Rab downregulated the Wnt pathway and impaired mucosal defense and regeneration. Microbiota involvement was indicated by fecal transplantation. Integrated metagenomic and metabolomic analyses revealed that Rab induced intestinal dysbiosis and reduced ileal bile acids, particularly 7-ketolithocholic acid (7KLCA) and chenodeoxycholic acid (CDCA). Faecalibaculum rodentium supplementation restored ISC self-renewal by converting CDCA to 7KLCA. In vitro, 7KLCA activated Wnt signaling to rescue Rab-induced stem cell impairment. In vivo, both 7KLCA and Gly-β-MCA (intestinal FXR antagonists) suppressed the FXR-FGF15 axis, restored the expression of hepatic bile acid synthesis enzymes, and promoted epithelial repair, thereby mitigating DSS-induced injury.<h4>Conclusions</h4>Chronic PPI use impairs ISC self-renewal by disrupting the microbiota-7KLCA-Wnt axis. F. rodentium or 7KLCA supplementation ameliorates PPI-induced effects, highlighting a microbe-metabolite axis as a pivotal mechanism and potential therapies for PPI-associated intestinal damage.

<h4>Background</h4>Proteomic profiling offers thorough insights into protein structure and function, as well as it acts as an essential approach for analyzing molecular changes at the tissue level. However, because of the proteome's diversity and dynamic nature, biomarker discovery remains challenging. By combining proteomics with bioinformatics, the level of understanding in relation to molecular interactions and disease processes can be improved. Through an integrative approach, few limitations can be addressed, thereby promoting proteomic profiling for the discovery of new therapeutic targets and novel biomarkers for a variety of disorders.<h4>Aim</h4>To identify differentially expressed protein markers and their key molecular pathways associated with Oral Potentially Malignant Disorders.<h4>Methods</h4>Systematic Review was conducted following the PRISMA guidelines and the protocol registered in the International Prospective Register of Systematic Reviews (PROSPERO) with the registration ID number CRD42024557545. A comprehensive literature review was performed using electronic databases, yielding 12,797, studies from which 15 eligible articles were selected. The Newcastle-Ottawa Scale was used to assess the risk of bias. Vote counting was performed to identify proteins reported in more than one study. A bipartite network was constructed using Cytoscape to identify shared and disease-specific protein markers. Lesion-wise protein-protein interaction networks were generated using STRING and analysed in Cytoscape to identify highly interconnected hub proteins, and pathway enrichment analysis for these hubs was performed using Reactome.<h4>Results</h4>A total of fifteen studies (Leukoplakia (LK) - n = 1, Proliferative Verrucous Leukoplakia (PVL) - n = 2, Oral Submucous Fibrosis (OSMF) - n = 7, and Oral Lichen Planus (OLP) - n = 5) were included. The Newcastle-Ottawa Scale was used to evaluate methodological quality and the quality of studies included in this systematic review was high for 4 articles and moderate in the remaining 11. The most commonly employed technique was mass spectrometry. A total of 318 candidate proteins (LK - 14, PVL - 82, OSMF - 172, and OLP - 50) were identified across the oral potentially malignant disorders. Key markers identified through vote counting included ERO1A, NUCB1, RHOA, and IL36A for PVL; LUM, KRT1, KRT9, ALB, and VIM for OSMF; and ALB, LYZ, HP, HBB, and AMY1A for OLP. The bipartite network showed that OSMF and OLP shared the highest number of proteins, indicating the strongest overlap among lesions. Network analysis further highlighted distinct hub proteins for each lesion: for LK- AMY1A, AMY1B and APOA1; for PVL- CFL1, RHOA and CDC42; for OSMF- HSP90AA1, ENO1 and SERPINA1; and for OLP- HP, B2M, and ORM1. Lesion-specific pathway enrichment revealed that LK was associated with epithelial differentiation, PVL with oncogenic signaling, OSMF with stress-driven fibrosis, and OLP with immune-mediated inflammation.<h4>Conclusions</h4>Proteomic expression offers insights into disease pathogenesis by identifying important molecular changes across OPMDs. However, the majority of biomarkers are still in the exploratory stage due to the considerable variation in lesion types, sample sources, proteomic techniques, and reporting systems. In order to create reliable and clinically applicable biomarkers, future studies should concentrate on combining multi-omics techniques with large-scale, standardized cohorts.

Neuroinflammation, oxidative stress and metabolic dysfunction are co-dependent drivers of neuropathic pain, jointly sustaining reactive gliosis and spinal sensitization. The interplay among neuroinflammation, redox imbalance, and metabolic reprogramming in glial cells represents a crucial process in this context, but the integrative mechanisms involved remain elusive. Poly(ADP-ribose) polymerase 1 (PARP1), a nuclear enzyme activated by genotoxic stress, has emerged as a key regulator of neuroinflammatory-associated diseases. The aim of the present study was to investigate the role of PARP1 in the pathophysiology of neuropathic pain and to evaluate the neuroglia phenotype and metabolism. Using a combination of in vitro glial cell cultures and an in vivo model of peripheral nerve injury, we demonstrated that sustained PARP1 activation triggers parthanatos, apoptosis signalling, and reactive gliosis, promoting neuroinflammation and maladaptive redox-inflammatory coupling in the spinal cord. At the behavioural level, PARP1 inhibition attenuated mechanical allodynia and improved motor coordination in rats with chronic constriction injury (CCI) of the sciatic nerve. Multi-omics profiling revealed a broad restoration of injury-altered proteomic and metabolic signatures, converging on restoring redox imbalance and related metabolic pathways, including glutathione and amino acid metabolic processing. Cell-type-specific knockdown of PARP1 established a causal link between redox balance, mitochondrial metabolism, and intercellular crosstalk during microglia inflammatory priming. Taken together, these findings highlight PARP1 as a functional regulator of the glial cell reactive phenotype in neuropathic pain and support its inhibition as a strategy to modulate central sensitization mechanisms.

4-Chlorophenoxyacetic acid (4-CPA), a synthetic auxin analog, is employed in agriculture both as a plant growth regulator and as a constituent of herbicide formulations. Consequently, the establishment of simple and rapid detection methods is essential for effective environmental monitoring. This study reports the first development of a homogeneous fluorescence polarization immunoassay (FPIA) for the determination of 4-CPA. The monoclonal antibody (M1), raised against 4-CPA, was evaluated as a recognition element. Furthermore, two fluorescently labeled 4-CPA tracers-with ethylenediamine fluorescein thiocarbamate and aminohexylaminocarbonylfluorescein-were synthesized and purified, and their structures were unequivocally confirmed by high-performance liquid chromatography coupled with high-resolution mass spectrometric detection (HPLC-HRMS). Optimal concentrations of monoclonal antibodies and tracers were established, yielding a limit of detection of 1.2 ng/mL. The assay demonstrated a broad dynamic range of 2.3-300 ng/mL and a rapid analysis time of 15 min. Validation via the standard addition method in authentic open water samples resulted in recovery rates of 98-112%. To address the cross-reactivity with the prevalent herbicide 2,4-dichlorophenoxyacetic acid (2,4-D), two novel strategies were devised and successfully implemented. The first approach involves the concurrent execution of two separate FPIAs-one for 2,4-D and one for 4-CPA-followed by the mathematical resolution of two analyte concentrations from the two measured binding values. The second strategy entails the preliminary selective removal of 2,4-D from sample matrices using affinity chromatography columns with immobilized anti-2,4-D antibodies prior to FPIA for 4-CPA. These proposed methodologies appear highly promising for overcoming the inherent limitations of traditional immunoassays when faced with significant cross-reactivity among structurally analogous compounds.

The canonical transsulfuration (TSS) pathway enzymes cystathionine β-synthase (CBS) and cystathionine γ-lyase (CTH) are traditionally recognized for their roles in the sequential conversion of homocysteine to cysteine and in endogenous hydrogen sulfide (H₂S) production. Increasing evidence, however, suggests that these enzymes may also exhibit non-canonical ("moonlighting") functions that extend beyond metabolic regulation. In this review, we evaluate the hypothesis that CTH may participate in translational regulation, particularly in the control of hypoxia-inducible factor-1α (HIF-1α) expression in clear cell ovarian carcinoma (CCOC). We first highlight limitations of the prevailing H₂S- and cysteine-centric view of the TSS pathway, which may not fully explain emerging context-dependent functions of CTH in cancer biology. Current evidence suggests that CTH enhances HIF-1α protein expression through mechanisms independent of transcription, protein stability, or H₂S production, implicating a potential role in translational regulation, although direct mechanistic evidence remains limited. To critically evaluate this emerging hypothesis, we categorize evidence according to its level of experimental support, ranging from direct experimental evidence to indirect mechanistic observations and computational predictions. Within this framework, we examine three non-mutually exclusive models: (1) regulation through PI3K/AKT/mTOR-dependent translational signaling; (2) modulation of translational control through interaction with translation-associated proteins and RNA-binding proteins (RBPs) involved in <i>HIF1A</i> mRNA regulation; and (3) the more speculative possibility of direct interaction between CTH and <i>HIF1A</i> mRNA. Collectively, these observations support a model in which CTH contributes to selective translational regulation beyond its canonical metabolic functions, potentially linking sulfur metabolism to stress-adaptive gene expression in cancer.

<i>Background and Objectives</i>: Neurological complications of SARS-CoV-2 infection frequently impair patients' long-term quality of life. This study aimed to identify clinical and laboratory risk factors-including inflammatory markers and micronutrients-for the occurrence or worsening of neurocognitive disorders in long COVID patients. <i>Materials and Methods</i>: In this prospective observational study, patients presenting with long COVID neurological manifestations were stratified by baseline MoCA score into two groups (≥23 and <23). Clinical, laboratory (inflammatory markers, 25-hydroxy vitamin D, vitamin B12, folic acid), and neuroimaging assessments (global cortical atrophy scale, Fazekas score) were performed over 24 months. Propensity score matching (PSM) for age, gender, and neurological comorbidities yielded 54 patients per group. <i>Results</i>: In the MoCA ≥ 23 group, significant predictors of cognitive decline included severe COVID-19 (OR = 2.211, 95% CI = 1.819-5.973, <i>p</i> = 0.012), autoimmune comorbidities (OR = 1.676, 95% CI = 1.191-2.390, <i>p</i> = 0.043), and elevated neutrophil-to-lymphocyte ratio (NLR; OR = 1.586, 95% CI = 1.431-2.122, <i>p</i> = 0.011). In the MoCA < 23 group, independent predictors were diabetes mellitus (OR = 3.021, 95% CI = 2.65-14.004, <i>p</i> = 0.016), autoimmune comorbidities (OR = 4.987, 95% CI = 1.412-6.033, <i>p</i> = 0.021), and NLR (OR = 5.944, 95% CI = 2.353-19.321, <i>p</i> = 0.015). Serum vitamin D levels were significantly associated with MoCA scores in both groups. <i>Conclusions</i>: COVID-19 severity, autoimmune comorbidities, NLR, and serum vitamin D represent key risk factors for neurocognitive decline in long COVID, highlighting potential targets for early intervention.

Mitophagy is a selective autophagy that degrades dysfunctional mitochondria to maintain cellular homeostasis. Mitophagy is functionally coordinated with and regulated by mitochondrial biogenesis and mitochondrial dynamics, which include mitochondrial fusion, mitochondrial fission, and mitochondrial trafficking. Furthermore, researches have demonstrated that mitophagy plays a critical role in the occurrence and development of digestive cancer. Nonetheless, the mechanism of how mitophagy modulates digestive cancer and the mechanism of how mitochondrial biogenesis and dynamics influence mitophagy warrant more investigations. This review summarizes the current understanding of the regulatory mechanism of mitophagy and outlines recent advances from investigations that explore how mitochondrial biogenesis and dynamics coordinate with mitophagy. Additionally, this review provides a comprehensive view about how mitophagy could regulate the occurrence and development of digestive cancer. A deeper understanding about the role of mitophagy in regulation of digestive cancer benefits the development of more efficient therapeutic strategies for patients.

Diabetes mellitus (DM) and neurological disorders are rapidly converging global health burdens, driven by population ageing, the growing prevalence of metabolic syndrome, and limited early detection and disease-modifying therapies for many neurological syndromes. Beyond its established role in diabetes-related peripheral neuropathy, DM is increasingly implicated as a modifier of risk, phenotype, and prognosis across a wide range of central and peripheral nervous system diseases. In this narrative review, we synthesize current epidemiological, clinical, genetic, and mechanistic evidence examining the relationship between DM and 10 clinically important neurological disorders: Alzheimer's disease (AD), vascular dementia (VaD), Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), chronic inflammatory demyelinating polyradiculoneuropathy (CIDP), multiple sclerosis (MS), myasthenia gravis (MG), and neuromyelitis optica spectrum disorder (NMOSD). Across these conditions, DM acts as a context-dependent disease modifier, increasing risk in some disorders, appearing protective or delaying onset in others, and influencing disease phenotype, progression, and treatment response. We highlight potential areas of mechanistic convergence, such as insulin resistance, inflammation, disrupted energy homeostasis, and genetic predisposition, alongside important divergences shaped by disease-specific pathology. We also discuss the clinical and translational implications of this interface, including diagnostic challenges, opportunities for improved risk stratification, and growing interest in repurposing antidiabetic therapies, particularly metformin, glucagon-like peptide-1 receptor agonists, and sodium-glucose cotransporter-2 inhibitors, for neurological benefit. As the global burden of diabetes and neurological disease escalates, it is crucial to better understand the interplay between metabolic dysfunction, neurodegeneration, and neuro-immune pathways. The integration of insights across diseases may inform prevention strategies and support the development of therapeutic interventions at the metabolic-neurological interface.

Spinal motor neurons are essential for translating neural activity into coordinated muscle contraction, yet defining their functional subtypes across development remains a persistent challenge. While embryonic patterning establishes the initial positional and molecular framework of motor neuron identity, substantial refinement continues during early postnatal life as intrinsic electrophysiological properties, synaptic connectivity, and neuromuscular interactions mature. A major limitation in the field is the lack of temporally stable and functionally validated molecular markers that can reliably distinguish motor neuron subtypes across developmental stages, particularly during neonatal maturation when subtype-specific physiological features are emerging. In this review, we synthesize classical developmental studies with recent advances in single-cell transcriptomics, chromatin accessibility profiling, and multimodal approaches linking gene expression with electrophysiological and anatomical features. Focusing on lumbar spinal motor neurons that underlie locomotor behavior, we discuss how transcriptional programs, activity-dependent mechanisms, and non-cell-autonomous signals converge to shape subtype-specific maturation trajectories. We propose that motor neuron subtype identity is best understood as a dynamic molecular and physiological state shaped by developmental timing, circuit context, and activity-dependent mechanisms, rather than as a fixed category defined by a single marker. From this perspective, early postnatal life represents a sensitive window of identity consolidation during which molecular programs and functional properties become aligned. Establishing temporally robust subtype markers and integrating molecular and physiological datasets will be essential for resolving motor neuron diversity and for improving our understanding of subtype-selective vulnerability in neuromuscular diseases. While this review emphasizes embryonic and early postnatal development, understanding how molecular subtypes stabilize in the adult spinal cord, despite ongoing activity-dependent physiological plasticity, remains an essential reference point for defining temporally robust motor neuron identities.