Gene Literature Dashboard

Cellular heterogeneity presents a significant challenge to cancer treatment. Antibody therapies targeting individual tumor-associated antigens can be extremely effective but are not suited for all patients and often fail against tumors with heterogeneous expression as tumor cells with low or no antigen expression escape targeting and develop resistance. Simultaneously targeting multiple tumor-specific proteins with multiple antibodies has the potential to overcome this barrier and improve efficacy, but relatively few widely expressed cancer-specific antigens are known. In contrast, neoepitopes, which arise from mutations unique to tumor cells, are considerably more abundant. However, since neoepitopes are not commonly shared between individuals, a patient-customized approach is necessary and motivates efforts to develop an efficient means to identify suitable target mutations and isolate neoepitope-specific monoclonal antibodies. Here, focusing on the latter goal, we use directed evolution in yeast and phage display systems to engineer antibodies from nonimmune, human antibody fragment libraries that are specific for neoepitopes previously reported in the B16F10 melanoma model. We demonstrate proof-of-concept for a pipeline that supports rapid isolation and functional enhancement of multiple neoepitope peptide-targeted monoclonal antibodies and demonstrate their robust binding to B16F10 cells and potent effector functions in vitro. These antibodies were combined and evaluated in vivo for anticancer activity in tumor-bearing mice, where they suppressed B16F10 tumor growth and prolonged survival. These findings emphasize the potential for clinical application of patient-customized antibody cocktails in the treatment of the many cancers poorly addressed by current therapies.

Lycopene is a carotenoid with strong antioxidant properties. Treatment with tripropylamine (TPA) greatly increases lycopene production by Blakeslea trispora, but the mechanism of this effect remains unclear. In this study, a reference-free transcriptome was used to analyze changes in gene expression in B. trispora after treatment with TPA. Downregulated genes were mostly involved in primary and secondary metabolic pathways, whereas upregulated genes were mostly related to autophagy and nucleotide excision repair. In addition, after TPA treatment, the mycelium swelling and the number of spores decreased. Our study suggests that TPA causes DNA damage, which upregulates the expression of nucleotide excision repair genes Xpb, Xpg, Csb, Polr2, and genome stability regulator gene Rmi1, as well as downregulation of Lsm2 and Dars2 and upregulation of G10, which are related to mRNA spliceosomes, leaving the cell in a state of starvation and triggering lysosomal-, peroxisomal-, and vacuole-autophagy. The autophagosome recycles material from spores, reduces spore production, and induces mycelial swelling, which, in turn, increases lycopene production and storage.

Polyglutamine (polyQ) diseases are monogenic fatal neurodegenerative disorders caused by a CAG repeat expansion that is translated into a toxic polyQ tract. There are nine polyQ diseases: Huntington's disease (HD), spinocerebellar ataxias 1, 2, 3, 6, 7 and 17 (SCA1, 2, 3, 6, 7, 17), dentatorubral-pallidoluysian atrophy (DRPLA) and spinal and bulbar muscular atrophy (SBMA). Although no disease-modifying therapies are available, lowering levels of the causative mutant polyQ protein is a promising potential treatment. Preclinically, the efficacy of polyQ protein-lowering compounds is often assessed using time-consuming Western blots (WB), which can produce variable results. Therefore, to improve throughput and accuracy of polyQ protein level quantification, Simple Western (SW) capillary immunoassays were developed. A panel of antibodies was screened for reactivity to the polyQ proteins on SW. The most promising antibodies were selected for further assay development. This resulted in optimised SW immunoassays for huntingtin (HTT), ataxin 1, 2 and 3 (ATXN1, 2, 3), atrophin 1 (ATN1) and androgen receptor (AR). Additionally, size-separation of the wild-type and polyQ-expanded mutant protein isoforms on SW was shown for ATXN1, ATXN3 and ATN1, allowing for their separate quantification. To facilitate size-separation of the larger HTT protein (≥ 348 kDa), a novel caspase 3-based assay was developed to generate N-terminal wild-type and mutant HTT fragments that could be separately quantified on SW in contrast to full-length HTT. In conclusion, SW capillary immunoassays were developed for polyQ proteins to improve preclinical research and aid the development of polyQ-lowering therapies for polyQ diseases.

Lung squamous cell carcinoma (LUSC) and lung adenocarcinoma (LUAD) are two major pathological types of non-small cell lung cancer (NSCLC), characterized by distinct patterns of lipid metabolism. However, the molecular mechanisms underlying lipid metabolism reprogramming specific to LUSC remain poorly understood. This study aims to fill this gap by identifying and characterizing EHHADH (enoyl-CoA, hydratase/3-hydroxyacyl CoA dehydrogenase) as a key regulator of medium-chain fatty acid metabolism in LUSC. The peroxisomal L-bifunctional enzyme is one of the important elements to control the peroxisomal fatty acid beta-oxidation pathway. Through high-expression genes related to lipid metabolism were identified by data mining, the expression and regulatory effects of EHHADH in different cell lines were investigated. EHHADH was highly expressed in LUSC cells and exhibited different expression patterns from those in LUAD cells. Knockdown of EHHADH in LUSC cell lines led to a marked reduction in cell proliferation. RNA sequencing following EHHADH silencing demonstrated significant changes in the expression of lipid metabolism-related genes in different cell lines, such as AZGP1, CAV1, CYP3A4, NR2F2, NR3C2, and RARG. Lipidomics analysis further demonstrated that EHHADH plays a crucial role in regulating intracellular and extracellular lipid profiles. EHHADH knockdown resulted in increased levels of long-chain fatty acids and storage lipids, while decreased levels of medium-chain fatty acids. Conversely, overexpression of EHHADH reduced long-chain fatty acids and storage lipids, while increasing specific medium-chain fatty acids. These metabolic alterations were consistent with changes in lipid metabolism-related protein expression, supporting the molecular mechanistic role of EHHADH in lipid regulation. In conclusion, EHHADH functions as an important regulator of lipid metabolism in LUSC and plays a key role in the occurrence, progression, and treatment of lung cancer. The important impact of EHHADH in lipid metabolism disorders suggests potential utility as a biomarker for diagnosis and a target for personalized treatment strategies in lung cancer.

<h4>Background</h4>Herpes Simplex Virus 1 (HSV-1) is a neurotropic virus causing encephalitis and post-infectious complications. Infections can induce a range of acute, subacute, and progressing brain disease, and in recent years it has emerged that immune responses are involved in the pathogenesis of these diseases.<h4>Methods</h4>Mice were infected with HSV-1 through corneal infection, and the brain stem was analyzed using single-cell and GeoMx spatial transcriptomics. Through these technologies we profiled temporal transcriptomic changes in cell populations, pathways, and cell-cell communication associated with antiviral activity and inflammation-induced disturbance of physiological brain structures and activities.<h4>Results</h4>We found that microglia proportions increased early after HSV-1 infection, followed by monocyte influx and later by T cells. The blood-brain barrier was disrupted, and transcriptomic profiles associated with homeostatic brain transcriptional activities were altered. Early transcriptional responses were dominated by antiviral and inflammatory activities. A microglia subpopulation with high type I interferon and chemokine expression localized to infection sites, likely mediating antiviral defense and immune recruitment. Monocyte subpopulations displayed a broader activation profile than microglia and was a central mediator of crosstalk between immune cells. Cytokines from microglia, monocytes, and T cells reprogrammed brain cells, notably endothelial cells and oligodendrocytes, disrupting brain functions. Comparing datasets from various brain diseases revealed the identified microglia subpopulation as specific to viral infections.<h4>Conclusions</h4>This study identifies a unique population of virus-activated microglia with antiviral and proinflammatory properties and reveals monocytes to be a key driver of interactions driving pathology in the virus-infected brain.

<h4>Background</h4>Intestinal regenerative capacity declines with age due to the declining intestinal stem cell (ISC) function. Dietary restriction (DR), the most robust anti-aging intervention to date, has been shown to improve ISC function in young mice.<h4>Objectives</h4>The goal of this study is to evaluate the effect of short-term DR (stDR) on ISC function when implemented late in life.<h4>Methods</h4>Two- and twenty-one-month-old male C57BL/6 mice, fed on National Institutes of Health-31 (NIH-31), an open-source, unpurified, natural ingredient diet, were subjected to ad libitum feeding or stDR (40% DR for 4-mo) with 6 mice/age/treatment. Effects of age and stDR on ISC (leucine-rich repeat-containing G-protein coupled receptor 5) function was evaluated using in vitro enteroid-forming assay followed by ribonucleic acid sequencing (RNA-seq) analysis (using strand next-generation sequencing analysis software) and oxidative whole-genome bisulfite sequencing for deoxyribonucleic acid (DNA) methylation (using the R package methyl kit). Statistical analysis was conducted, where appropriate, using a 1-way analysis of variance with multiple testing corrections or Student's t-test.<h4>Results</h4>Our data shows that stDR increases enteroid formation when implemented late in life by ≥10-fold (P ≤ 0.05). RNA-Seq analysis identified 38 genes not commonly linked to ISC function but showed a 2-fold change (P ≤ 0.05) with age, which was prevented by stDR. Finally, evaluation of the methylation status of selected genes from RNA-seq (Slc28a3, Ly9, Cd2, Platr4) showed changes in methylation at Cytosine-Guanine (CG) sites in the cis-acting regions of the genes with age which were also prevented by stDR (P ≤ 0.05).<h4>Conclusions</h4>Our study shows for the first time that late-life stDR can improve the function of leucine-rich repeat-containing G-protein coupled receptor 5 ISCs from old mice. These data suggest that late-life nutritional strategies might be effective in improving gut function that declines with age and reduce the risk of age-related intestinal disorders. Our study also provides the first data showing that DNA methylation may play a role in the ability of DR to improve ISC function.

Recently, research on the competing endogenous RNAs (ceRNAs) in cancer has been in full swing, emphasizing their importance as critical RNAs in cancer progression. Enhancer of zeste 2 polycomb repressive complex 2 subunit (<i>EZH2</i>) is a ceRNA that has been introduced as a potential therapeutic target in many cancers. Due to EZH2's dual role as an oncogene and tumor suppressor in cancer, a more thorough exploration of its ceRNA functions may enhance clinical cancer treatment approaches. In the current scoping review, we searched several online databases to identify experimentally validated ceRNA axes, including <i>EZH2</i> in human cancers. We identified 66 unique axes consisting of 30 microRNAs (miRNAs), 32 long non-coding RNAs (lncRNAs), 9 messenger RNAs (mRNAs), and 14 circular RNAs (circRNAs). Notably, <i>SPRY4-IT1 -</i> miR-101-3p - <i>EZH2</i> and <i>XIST -</i> miR-101-3p - <i>EZH2</i> were recurrent axes observed in multiple cancer types. Among the most frequent miRNAs were miR-101-3p, miR-144-3p and miR-124-3p, and ceRNAs including <i>SPRY4-IT1</i>, <i>XIST</i>, <i>SNHG6</i>, <i>HOXA11-AS</i>, <i>MALAT1</i>, and <i>TUG1</i> emerged as frequent competitors of <i>EZH2</i> for miRNA binding. This scoping review highlights the diversity of <i>EZH2</i>-containing ceRNA axes in cancer, suggesting their potential as therapeutic targets. Further studies are needed to clarify their roles and clinical utility.

Runs of homozygosity (ROHs) are caused by identical haplotypes inherited from ancestors. ROHs provide useful information regarding the inbreeding rate, demographics, and selection history. The Yeonsan Ogye (YO) breed is an indigenous chicken in Korea that is characterized by a completely black body. In this study, we investigated ROH in the YO genome to determine ROH-based inbreeding coefficients and their correlations with other inbreeding estimators, then analyzed their genetic characteristics. Using 600K single nucleotide polymorphism (SNP) chip information for 189 chickens, we found 20,339 ROHs in the YO population. The average number of ROHs was 107, the total average ROH length was 165 Mb, and the average ROH length was 1.542 Mb. Most ROHs were short (< 8 Mb), suggesting a past population bottleneck. The average inbreeding coefficient (<i>F_ROH</i> ) calculated based on ROHs was 0.184 and this was correlated with other inbreeding coefficients estimated using allele frequencies. 17 ROH islands were detected and these regions exceeded the threshold of the top 1% of SNPs among SNPs present in ROHs. In the ROH islands, 152 genes were annotated, some of which were genes associated with meat production traits and hyperpigmentation in chickens. A comparison of overlapping regions between ROH islands and quantitative trait loci (QTLs) indicated that most QTLs were related to color traits. These results will help to optimize conservation strategies for the YO breed.

Testicular orphan receptors TR2 and TR4 serve as central regulators of erythropoiesis, orchestrating the entire continuum of erythroid progenitor cell proliferation, differentiation, and maturation. As core components of the direct repeat erythroid determinant (DRED) complex, they activate erythroid-specific transcriptional programs to dynamically control the spatiotemporal expression of globin genes. These nuclear receptors not only engage in functional interactions with key erythroid transcription factors GATA1 and KLF1 to coregulate erythroid differentiation and maturation but also recruit epigenetic modifier complexes such as DNMT1 and LSD1 to modulate chromatin states dynamically. Research has established that dysfunctions in TR2/TR4 are implicated in β-thalassemia and sickle cell disease (SCD): β-thalassemia is associated with the defective silencing of γ-globin genes, while in SCD, TR2/TR4 antagonizes BCL11A to reactivate fetal hemoglobin (HbF) expression. This review systematically dissects the molecular regulatory networks of TR2/TR4 in erythroid cells, interprets their dual regulatory properties across different stages of erythroid differentiation, and explores the therapeutic potential of targeting TR2/TR4 for treating erythroid-related disorders such as β-thalassemia and SCD, thereby providing novel directions for hematological disorder therapy.

<b>Background:</b> The relationship between oral contraceptive (OC) use and pancreatic cancer (PC) risk remains controversial, with inconsistent findings reported in observational studies. To clarify this relationship and better identify potential risk factors for PC prevention, more unbiased and robust approaches are needed. <b>Methods:</b> We investigated the potential causal relationship between OC use and PC risk using a two-sample Mendelian randomization (MR) analysis, with blood protein quantitative trait loci (pQTLs) as instrumental variables. To ensure the robustness of our findings, we performed a series of sensitivity analyses, colocalization analyses, and reverse MR. The causal effects of protein-coding genes on PC risk, as well as their expression patterns across different single-cell types, were subsequently investigated. To elucidate the potential pathogenic pathways, we conducted pathway enrichment analysis, protein-protein interaction (PPI) network analysis, and causal inference. <b>Results:</b> Our MR analysis identified five drug-targeted proteins significantly associated with PC risk. Higher levels of COMT, AGT, FN1, and UGT1A1, as well as lower levels of SERPINC1, were associated with an increased risk of PC. Among these, AGT, FN1, and COMT demonstrated consistent associations across sensitivity analyses and downstream analyses, providing robust evidence supporting their involvement in PC risk. <b>Conclusions:</b> This study provides genetic evidence suggesting, in European groups, a potential causal link between OC use and increased PC risk, possibly mediated through drug-targeted proteins such as AGT and FN1. These results highlight the importance for further research to elucidate the underlying mechanisms and assess the implications of OC use on PC risk.

Calcium pyrophosphate dihydrate deposition (CPPD) disease, also known as pseudogout, is the most common cause of calcification of ligamentum flavum (CLF) compared to other degenerative conditions. Symptomatic CLF is a rare cervical spine disorder that leads to spinal cord compression, resulting in myelopathic symptoms. We report two rare cases of pseudogout-induced cervical myelopathy, along with a review of the literature. Both patients were seen at Al-Azhar University Hospital in Cairo, Egypt, presenting with severe neck pain, gait disturbances, and hand clumsiness. The first patient had these symptoms for two months, while the second patient experienced them for three months. The second patient also had restricted motion in extension and rotation, as well as shooting pain in both upper extremities. Cervical spine MRI revealed posterior cord compression and myelomalacia at the C3-C6 levels in the first patient and at the C4-C5 levels in the second patient. Pseudogout (CPPD) disease was suspected as the cause of spinal cord compression, and histopathological analysis of the deposits found at the site of compression during the decompression procedure confirmed the diagnosis. The deposits were characterized by rhomboid blue calcium crystals that were mildly birefringent, distinguishing them from the needle-shaped crystals seen in gout. At the four-week postoperative follow-up, both patients showed significant improvement in clinical and functional outcomes, as measured by the Japanese Orthopaedic Association score. In summary, CLF due to pseudogout (CPPD deposition) is a rare but clinically significant cause of cervical myelopathy. Histopathological examination is crucial for a definitive diagnosis. Early posterior cervical decompression with instrumentation appears to be an effective treatment, though larger studies and long-term follow-up are necessary to confirm these findings and optimize management strategies.

<h4>ABSTRACT</h4> A major challenge in melanoma treatment is the emergence of aggressive metastases from previously dormant disseminated cancer cells (DCCs). Here we identify Midkine (MDK) as a key factor that disrupts melanoma DCC dormancy by triggering an autocrine signal that suppresses the dormancy inducer NR2F1 and alters the p-p38/p-ERK ratio, via ALK signaling. Dormant melanoma DCCs in lymph nodes and visceral tissues exhibit an MDK LOW /NR2F1 HIGH phenotype. Transient NR2F1 activation for one month with a potent agonist counteracts MDK-driven mTORC1 activity and suppresses lung metastasis in mice bearing human melanoma DCCs. Dual targeting of MDK (genetic blockade) and NR2F1 (activation) markedly limits metastatic outgrowth and extends survival for 6 months in mouse models independent of BRAF status. These findings support the MDK-NR2F1 axis as a promising therapeutic target to sustain dormancy and prevent melanoma recurrence. <h4>Statement of Significance</h4> This work identifies Midkine as a key disruptor of tumor dormancy in melanoma, driving metastatic awakening via NR2F1 suppression. Dual targeting of Midkine (inhibition) and NR2F1 (activation) prolongs survival in preclinical models.

The Hippo signaling pathway, so named for its massive overgrowth mutant phenotypes, has become one of the most exciting signaling pathways to emerge in the field of reproductive biology. While disruption of Hippo is associated with tumorigenesis in many organs and tissues, relatively less is understood about the normal roles of Hippo signaling in the reproductive organs. Here, we highlight the recent literature illuminating the roles of Hippo pathway members in mouse and human reproduction. We place special emphasis on the inputs and outputs of Hippo signaling during preimplantation development, where Hippo signaling has been extensively studied in both mouse and human. We note a common emerging theme is the critical and highly conserved role of Hippo signaling in epithelia of the reproductive organs. We also discuss human reproductive disorders, whose etiology may be related to dysregulation of Hippo signaling, and possible therapies that have been proposed to correct this dysregulation. Finally, we describe the edge of our knowledge, which currently limits our understanding of Hippo signaling in reproductive health and disease.

<h4>Abstract</h4>Hydrogen sulfide (H 2 S), an important gaseous signaling molecule, plays a critical role in maintaining vascular homeostasis. H 2 S participates in numerous biological functions, including redox regulation, interactions with other signaling molecules, and post-translational modifications of proteins through sulfhydration. Additionally, H 2 S influences key pathological processes such as inflammation, oxidative stress, and cell apoptosis. Dysregulation of endogenous H 2 S metabolism has been closely linked to the development of various vascular diseases, including aortic aneurysms, aortic dissection, atherosclerosis, and thrombotic conditions. Various endogenous and exogenous H 2 S donors have been developed, and these donors have demonstrated promising effects in preclinical models of vascular diseases such as atherosclerosis, pulmonary hypertension, and thrombosis by modulating oxidative stress, inflammatory pathways, and vascular remodeling. This review consolidates the current knowledge on the effects of H 2 S on vascular function and offers a comprehensive summary of recent advancements in the development and application of H 2 S donors in vascular disease research.

Lung injury in preterm infants leads to structural and functional respiratory deficits, with a risk for bronchopulmonary dysplasia (BPD) that in its most severe form is accompanied by pulmonary hypertension (PH). To identify potential cellular and molecular drivers of BPD in humans, we performed single-cell RNA sequencing of preterm infant lungs with evolving BPD and BPD + PH compared to term infants. Examination of endothelial cells reveals a unique, aberrant capillary cell-state in BPD + PH defined by ANKRD1 expression. Within the alveolar parenchyma in infants with BPD/BPD + PH, predictive signaling analysis identifies surprising deficits in the semaphorin guidance-cue pathway, with decreased expression of pro-angiogenic transcription factor FOXF1. Loss of semaphorin signaling is replicated in a murine BPD model and in humans with causal FOXF1 mutations for alveolar capillary dysplasia (ACDMPV), suggesting a mechanistic link between developmental programs underlying BPD and ACDMPV and uncovering a critical role for semaphorin signaling in normal lung development.

The MYC oncoprotein regulates numerous genes involved in processes such as cell cycle control and mitochondrial and ribosomal structure and function. This requires heterodimerization with its partner, MAX, and binding to specific promoter and enhancer elements. Here, it is shown that MYC and MAX also bind near transcriptional end sites (TESs) of over one-sixth of all annotated genes. These interactions are dose-dependent, evolutionarily conserved, stabilize the normally short-lived MYC protein and regulate expression both in concert with and independent of MYC's binding elsewhere. MYC's TES-associated binding, occurring in coordination with other transcription factors, alters the chromatin landscape, increases nuclease susceptibility and alters transcriptional read-through, particularly in response to certain stresses. MYC-bound TESs can directly contact promoters and appear to fine-tune gene expression in response to both physiologic and pathologic stimuli. Collectively, these findings support a previously unrecognized role for MYC in regulating transcription and its read-through via direct intragenic contacts between TESs and promoters.

Osteoarthritis (OA) associated with obesity is increasingly recognized as a distinct phenotype, driven by lipid metabolic imbalance and related inflammation. A particularly troublesome issue is that even after successfully correcting obesity, OA progression and lipid metabolic imbalance persist within the joint microenvironment, suggesting local lipid metabolism regulation as a potential treatment option. G-protein-coupled receptor 120 (GPR120), a primary receptor for long-chain fatty acids (including docosahexaenoic acid, DHA), has recently been found to play a pivotal role in regulating lipid homeostasis and suppressing inflammation. Here, we present ChD-FL/sgGPR, enabling dual endogenous-exogenous GPR120 activation. ChD-FL/sgGPR is a chondrocyte-biomimetic, fluorinated phenylboronic acid (FPBA)-modified ionizable liposome that codelivers DHA and a CRISPRa system comprising GPR120-specific sgRNA (single guide RNA) and dCas9-VPR mRNA (dead Cas9 fused to VP64-p65-Rta activator domain). Specifically, FPBA modification of liposomes enhances lysosomal escape and nuclear entry of RNA, while coextrusion with chondrocyte membranes facilitates cartilage-targeted delivery. In the coculture system of adipocytes and OA chondrocytes, ChD-FL/sgGPR significantly boosts chondrocytes GPR120 expression, facilitates lipid clearance via PPARγ signaling, and diminishes inflammatory mediators. In obese rat OA models, intra-articular injection of ChD-FL/sgGPR prolongs local retention, inhibits cartilage catabolism, and mitigates subchondral bone deterioration, collectively decelerating OA progression. By integrating CRISPR-mediated gene upregulation with DHA-induced receptor stimulation, this platform rebalances lipid metabolism in OA cartilage, offering a promising, mechanism-driven therapy for obesity-associated OA.

<h4>Objective</h4>To examine the expression of non-SMC condensin II complex subunit D3 (NCAPD3) in papillary thyroid carcinoma (PTC) tissues, assess its impact on the growth and metastatic potential of PTC cells, and investigate its role in regulating glycolysis to uncover the underlying mechanisms involved.<h4>Methods</h4>NCAPD3 levels in PTC tissues were detected using immunohistochemistry. siRNA transfection was used to silence NCAPD3 in K1 and TPC-1 cells. Cell proliferation rates were detected using the Cell Counting Kit-8 assay, migration rates were evaluated using wound healing and Transwell cell migration assays, and invasion rates were assessed using the Transwell-Matrigel cell invasion assay. Moreover, the aerobic glycolysis-related factors lactate, lactate dehydrogenase A (LDHA), and pyruvate kinase M2 (PKM2) were detected using kits.<h4>Results</h4>NCAPD3 was highly expressed in PTC tissues. Its expression showed no significant association with patient age, gender, or lymphocytic thyroiditis but was significantly correlated with larger tumor size and lymphovascular invasion. NCAPD3 expression significantly decreased in K1 and TPC-1 cells after transfection with siRNA. Low NCAPD3 expression reduced the proliferation rate of K1 and TPC-1 cells and inhibited cell migration and invasion. Moreover, NCAPD3 silencing decreased LDHA, PKM2, and lactate levels.<h4>Conclusions</h4>NCAPD3 was highly expressed in PTC tissues, and correlated with aggressive features (tumor size and lymphovascular invasion). NCAPD3 silencing inhibited proliferation, migration, invasion, and aerobic glycolysis of PTC cells. Therefore, NCAPD3 may serve as a potential therapeutic target for PTC.

<h4>Abstract</h4>Polymicrogyria (PMG) represents a complex disorder involving malformation of the cortex. There have not been any comprehensive genetic studies of PMG from India. Here, we have done a whole-exome sequencing (WES) study of the PMG patients in a South Indian population to identify the genetic causes of PMG. The study design was descriptive. Twenty patients with PMG were recruited for the study. WES was done for all the participants. Our study identified a few potential candidate genes associated with PMG, including ROS1, PIK3R2, SUSD2, NPIPB15, RBMX, DENND4B, KRT18, PUS1 , and TTC28 . Notably, some of these genes have been identified as having a substantial role in neurodevelopment. Some novel candidate genes of PMG were also identified in this study. The PMG-associated genes were enriched in the biological processes involved in cell adhesion, cytoskeleton organization, and nervous system development.

Identifying genetic mutations contributing to solid tumors by altering the biological pathways related to tumor formation and development is essential for the development of targeted therapies. This study aimed to identify commonly mutated genes and altered pathways in canine solid tumors. Four dogs with different types of naturally occurring neoplasias (urothelial carcinoma, adenocarcinoma, rhabdomyosarcoma, and chondrosarcoma) were randomly selected and classified into carcinoma and sarcoma groups based on histopathological findings. Tumor tissues were analyzed using whole-genome sequencing, and significant variants shared within each tumor group were identified. Gene set enrichment analyses were conducted to compare the biological and functional pathways altered by the mutations in each carcinoma and sarcoma group. Forty-three and fifty-eight genes were identified in the carcinoma and sarcoma groups, respectively. Distinctions between the two tumor groups were noted for mutations related to tumor metastatic function. Mutations were identified in genes encoding cell adhesion molecules in the carcinoma group, whereas significant variations in extracellular matrix-related molecules were evident in the sarcoma group. This study revealed mutations and modified pathways associated with immune and tumor metastatic functions in canine carcinoma and sarcoma, indicating their significant relevance to the development and progression of each tumor group. Additionally, the distinctions indicated that different therapeutic approaches were required for each tumor group.

Cervical cancer remains one of the leading causes of cancer-related mortality among women worldwide, particularly in low- and middle-income countries, highlighting the need for improved strategies in treatment and management. This study aimed to investigate the anti-cervical cancer potential and molecular mechanisms of Solanecio mannii (S. mannii) aqueous extract using a "multi-compound, multi-target, multi-pathway" approach, integrating both computational and experimental methods. The metabolomics profile of the extract was analysed, and its selective cytotoxicity was assessed against human cervical cancer cell lines (HeLa cells) using the CCK8 assay. A network pharmacology approach identified potential molecular targets and pathways, which was complemented by molecular docking and dynamic simulation. The expression levels of key targets were validated experimentally using quantitative real-time polymerase chain reaction. Additionally, the extract's effects on apoptosis, autophagy, and cell cycle progression were studied experimentally. The aqueous roots extract exhibited selective cytotoxicity against HeLa cells with an IC50 of 12.53 ± 4.983 μg/ml. The network pharmacology analysis identified 25 drug-like compounds targeting 493 unique cervical cancer-associated proteins, forming a protein-protein interaction network of 465 nodes and 2230 edges, and implicated in 178 enriched KEGG pathways. Key targets, including NFΚB1, PIK3CA, HIF1A, STAT3, HSP90AA1, HSP90AB1, PPARG, and ESR1 were experimentally downregulated. Furthermore, S. mannii aqueous roots extract triggered apoptosis through endoplasmic reticulum stress, DNA damage, and activation of the non-transcriptional, P53-mediated mitochondrial apoptotic pathway. Additionally, the extract inhibited hypoxia and autophagy, and induced cell cycle arrest at the G2/M phase, even in the presence of oncogenic HPV proteins (E6 and E7). In conclusion, Solanecio mannii aqueous roots extract demonstrates a "multi-compound, multi-target, multi-pathway" molecular mechanism against cervical cancer.

CD4⁺ T cells are indispensable for optimal immunity to <i>Mycobacterium tuberculosis</i> (<i>M.tb</i>), a pathogen that triggers tuberculosis (TB) in humans. <i>M.tb</i>-specific human CD4⁺ T cells are known to polarize toward an interferon-γ (IFN-γ)-producing, CCR4^-CCR6⁺CXCR3⁺T-bet⁺RORγT⁺ T helper 1* cell (T_H1*cell) memory phenotype. We report that autosomal recessive deficiency of the human lymphocytic surface receptor LY9 (SLAMF3 and CD229), which is found in less than 10^-5 individuals in the general population, underlies TB in three unrelated patients due to selective impairment in IFN-γ production by T_H1* cells. T_H1* cells express higher levels of LY9 than other CD4⁺ T cells. Mechanistically, LY9 polarizes naïve CD4⁺ T cells toward memory T_H1* cells by inducing T-bet via signaling lymphocytic activation molecule (SLAM)-associated protein (SAP) and RORγT (thymus-specific retinoid-related orphan receptor γ) without SAP. LY9 costimulation enhances TCR-driven IFN-γ production of memory T_H1*, but not T_H1, cells in a T cell-intrinsic manner via NFAT1 (nuclear factor of activated T cells 1) and RORγT. LY9 is likely to govern an optimal T_H1* cell- and IFN-γ-dependent protective immunity to <i>M.tb</i> in humans.

Glycosaminoglycans (GAGs) are linear, highly acidic polysaccharides that serve as essential extracellular matrix components. There has been increasing evidence that GAGs can release gadolinium ions from complexes of magnetic resonance imaging contrast agents. This unintended release of gadolinium might be an initial step leading to gadolinium deposition disease, as observed in some patients after intravenous injection of such contrast agents. However, the molecular details of the release remain poorly understood. In this work, we provide direct evidence for gadolinium binding by GAGs using synthetic model substance Fondaparinux (FPX), a heparin mimetic. We observed FPX-gadolinium complexes in mass spectrometry experiments and electron paramagnetic resonance spectroscopy (EPR) and characterized the binding by EPR, isothermal titration calorimetry, and gas-phase infrared (IR) spectroscopy. Finally, we were able to follow the transchelation process on a molecular level by utilizing collision-induced dissociation experiments.

Infective endocarditis, a life-threatening condition, poses challenges for early diagnosis and personalized treatment due to insufficient biomarkers and limited understanding of its pathophysiology. Here, we performed proteomic profiling of plasma and vegetation samples from 238 patients with infective endocarditis and 100 controls, with validation in two external plasma cohorts (n = 328). We developed machine learning-based diagnostic and prognostic models for infective endocarditis, with area under the curve values of 0.98 and 0.87, respectively. Leucine-rich alpha-2-glycoprotein 1 and NADH:ubiquinone oxidoreductase subunit B4 are potential biomarkers associated with infection severity. Pathologically, protein networks characterized by glycometabolism, amino acid metabolism, and adhesion are linked to adverse events. Liver dysfunction may exacerbate the condition in patients with severe heart failure. Neutrophil extracellular traps emerge as promising therapeutic targets in Streptococcus or Staphylococcus aureus infections. Our findings provide insights into biomarker discovery and pathophysiological mechanisms in infective endocarditis, advancing early diagnosis and personalized medicine.

The prevalence of recessive disorder carriers among Vietnamese women is still indistinct. This study aims to assess the prevalence of carriers for common autosomal recessive and X-linked conditions among Vietnamese pregnant women and to identify common mutations within these genes. A cross-sectional study was conducted with 8,464 Vietnamese pregnant women with indications for carrier screening tests for recessive disorders from November 2022 to August 2023 at the Institute of DNA Technology and Genetic Analysis. The survey includes demographic information, and the genetic screening was conducted using next-generation sequencing (NGS) techniques, focusing on 13 specific recessive conditions. 8,464 Vietnamese pregnant women's records were involved in this study. 1,928 of them carried at least one genetic recessive condition, representing the frequency of a recessive disorder was 22.8%. The highest recessive disorders rate among pregnant women was found for the G6PD gene mutation (G6PD deficiency) at a rate of about 1 in 20 individuals, followed by the HBA1 and HBA2 gene mutations (Alpha Thalassemia) at a rate of about 1 in 25. Other common recessive carrier genes included SRD5A2 (5-alpha reductase deficiency) at a rate of about 1 in 27, HBB (Beta Thalassemia) at a rate of about 1 in 28, ATP7B (Wilson's disease) at a rate of about 1 in 40, PAH (Phenylketonuria) at a rate of about 1 in 40, and SLC25A13 (Citrin deficiency) at a rate of about 1 in 45. The prevalence of recessive carriers among Vietnamese pregnant women is high, and at least 1 in 5 pregnant women carries one recessive gene. It is essential to encourage Vietnamese pregnant women to conduct recessive carrier screening tests to reduce mortality rates among children and to implement effective pregnancy planning and childbirth.

GULP PTB domain containing engulfment adaptor 1 (GULP1) protein is involved in regulating biological processes such as endocytosis and apoptosis. The function of GULP1 in cancer, however, has been the subject of fewer studies; its importance as a potential prognostic factor in pancreatic cancer is still uncertain. Thus, the purpose of this work was to investigate GULP1's immunologic and oncogenic activities in a range of malignancies, as well as any potential relevance to pancreatic cancer. Using multiple bioinformatic databases, GULP1 expression, prognostic significance, mutation status, methylation and phosphorylation levels, biological functions, immune cell infiltration and immunotherapeutic responses and drug sensitivity were comprehensively assessed in pan-cancer and functionally validated in pancreatic cancer. The results revealed that GULP1 was differentially expressed in most tumours and correlated with poor prognostic indicators in most tumours, which may be related to the fact that GULP1 is involved in the regulation of apoptotic pathways. In addition, the differential expression of GULP1 was linked to immune cell infiltration levels, immunotherapy response, and chemotherapy resistance. GULP1 could hinder the body's ability to fight tumors and respond to immunotherapy by promoting the accumulation of immune cells and suppressing the activity of cytotoxic T lymphocytes. In pancreatic cancer, down-regulation of GULP1 expression inhibits proliferation, invasion and migration of pancreatic cancer cells. These phenotypic changes may be achieved by regulating HIPPO, mTOR, and RTK signaling pathways. Taken together, it makes sense to think that GULP1 could be a biomarker for immunotherapy and prognosis in pan- and pancreatic cancer.

Sarcopenia is defined as a progressive decrease in skeletal muscle mass and strength due to ageing, and is associated with adverse outcomes such as falls, fractures, and physical disability, leading to increased healthcare expenditures and mortality risks. Although sarcopenia has recently become a major challenge, there are currently no effective interventions or drugs marketed for sarcopenia. Therefore, new biomarkers for early diagnosis and targeted drugs for the treatment of sarcopenia are urgently needed. In the present research, transcriptomics and proteomics analyses were combined and experiments at the transcriptional and protein levels were conducted to identify key genes and molecular mechanisms underlying sarcopenia in senescence-accelerated mouse prone 8 (SAM-P8) mice, followed by molecular docking to predict targeted drugs. The combined omics analysis identified 8 key genes, while the experiments verified that only CD9 significantly decreased in sarcopenia. The gene set enrichment analysis (GSEA) results suggested that CD9 involved in ATP biosynthesis, mitochondrial biogenesis, and oxidative phosphorylation. Besides, dapoxetine, levomilnacipran, and milnacipran were predicted to target CD9 through molecular docking. Our study reported for the first time that CD9 is a novel potential biomarker of sarcopenia, and targeting CD9 may be a new idea for the development of therapeutic drugs for sarcopenia.

<h4>Background</h4>Cholangiocarcinoma (CHOL) is the second largest malignant tumor in the bile duct, only after primary liver cancer. Their high invasiveness can lead to poor prognosis. Over the past decade, research on CHOL has been ongoing, but there have been no breakthrough advancements. According to the literature, NCAPG2 is associated with the progression of various tumors, but its relevant role and value in CHOL have not been extensively studied.<h4>Methods</h4>We studied the expression of NCAPG2 in CHOL and its clinical and pathological diagnostic value based on TCGA and GEO datasets. We evaluated the association between NCAPG2 and immune-related cells in CHOL using the TISIDB database.<h4>Results</h4>We found that NCAPG2 has a higher expression level in CHOL and diagnostic significance in patients with CHOL. Additionally, we observed a correlation between immune cell infiltration and NCAPG2 levels in CHOL. Furthermore, NCAPG2 and its co-expressed genes were predominantly concentrated in ATP-dependent activity, DNA-related functions, and DNA helicase activity. The expression level of NCAPG2 is intricately linked to the pathological classification of CHOL.<h4>Conclusion</h4>Current research indicates that NCAPG2 may provide effective assistance for the early diagnosis and pathological classification of CHOL, potentially regulating the immune microenvironment and thereby influencing the occurrence and progress of CHOL.

<h4>Background</h4>Extracorporeal membrane oxygenation (ECMO) is increasingly used in patients after cardiac surgery; however, anticoagulation management has consistently been challenging. This study aimed to explore the feasibility of a low-anticoagulation strategy for post-cardiotomy ECMO (PC-ECMO).<h4>Methods</h4>A retrospective comparison was performed between two anticoagulation targets in adult patients undergoing veno-arterial ECMO after cardiac surgery at the Beijing Anzhen Hospital (Beijing, China) between January 2018 and November 2023. The low-anticoagulation (LAC) strategy group consisted of patients with an activated partial thromboplastin time (APTT) ratio of 1-1.5, whereas the conventional anticoagulation (CAC) strategy group included those with an APTT ratio of 1.5-2.5. The primary outcome was thrombotic complications associated with ECMO. Secondary outcomes included bleeding events, pulmonary infection, need for renal replacement therapy, in-hospital mortality rate, ECMO support duration, hours of mechanical ventilation, anticoagulation fraction, length of hospitalization, and transfusion volume(s).<h4>Results</h4>The study included data from 203 patients, who were divided into two groups: LAC (n = 108 [53.2%]) and CAC (n = 95 [46.8%]). Propensity score matching was used to balance confounding variables. A total of 43 patient pairs were successfully matched, and no significant difference was observed in thrombotic complications between the LAC and CAC groups (30.2% versus [vs.] 25.3%, respectively; p = 0.810). Meanwhile, no significant differences were observed in secondary outcomes and subgroups within the matched cohort, except for ECMO support time, which was shorter in the LAC group (119.6 h vs. 146.0 h; p = 0.015).<h4>Conclusion</h4>The low-anticoagulation strategy was feasible for PC-ECMO support.

<h4>Background</h4>Sirolimus, the therapy choice for lymphangioleiomyomatosis (LAM), displayed cytostatic but not cytocidal action, with disease recurrence after withdrawal. The aim of this study is to identify novel potential biomarkers and therapeutic strategies for LAM patients.<h4>Methods</h4>TMT-labeling proteomics was utilized for screening the differentially expressed proteins (DEPs) in the plasma of 10 LAM patients and 6 controls. Plasma levels of transferrin (TRF), ferritin (FRT) and beta2-microglobulin (B2M) were validated in a cohort of 30 LAM patients and 20 controls. The diagnostic efficacy of TRF with/without VEGF-D was assessed using ROC curve analysis. The therapeutic effects of a ferroptosis inducer artesunate (ART) were evaluated both in vitro Tsc2 - / - MEFs cells and in xenograft LAM models.<h4>Results</h4>Proteomics analysis revealed 132 DEPs between LAM patients and controls, which primarily enriched in the regulation of iron ion transport. LAM patients had decreased TRF, elevated FRT and B2M levels compared with controls in the confirmation cohort (p = 0.0386, p = 0.0327 and p = 0.0155, respectively) which independent with VEGF-D level or rapamycin therapy. TRF positively correlated with both FEV₁% predicted (r = 0.4486, p = 0.0251) and DLCO% predicted (r = 0.4018, p = 0.0516) of LAM patients. The combination of TRF and VEGF-D showed superior diagnostic value compared to individual indicator. ART induced the ferroptosis and inhibited the growth in Tsc2 - / - MEFs cells. In LAM animal models, ART exerted anti-tumor effects without obvious adverse effect.<h4>Conclusions</h4>LAM patients exhibit abnormal iron metabolism independent of VEGF-D level. Ferroptosis inducer ART holds promise as a therapeutic novel approach for treating LAM.

Chronic stress, a common risk factor for psychiatric disorders, is also implicated in the pathogenesis of Alzheimer's disease (AD). However, its underlying mechanisms remain elusive. Here, we provide evidence for chronic restraint stress (CRS), a widely used stress model in rodents, to regulate AD pathology. CRS not only induces prolonged depressive-like behaviors and cognitive deficits in young adult wild type (WT) mice, but also exacerbates a series of AD-related phenotypes in APP/PS1 mice, including impaired spatial learning and memory, increased β-amyloid plaques, promoted glial cells (astrocyte and microglial cell) activation and decreased dendritic spines in CA1 neurons. Single-nucleus RNA-sequencing analysis in hippocampus shows remarkable transcriptional changes in many cell type(s), and identifies oxidative phosphorylation pathway, a major source for adenosine triphosphate (ATP) production, is significantly downregulated in CA1 neurons by CRS stimuli. Furthermore, dysfunctional mitochondria and reduced ATP levels are also observed in CA1 neurons of CRS exposed WT and APP/PS1 mice. Interestingly, infusion of ATP in CA1 region abolishes the deficits in cognition, dendritic spines and glial activation in CRS exposed APP/PS1 mice. Taken together, these results uncover an unrecognized function of CA1 neurons in regulating CRS induced AD pathologies, and suggest ATP as a promising therapeutic strategy to improve brain health under stress condition.

<h4>Background</h4>Cardiac procedures, particularly those requiring cardiopulmonary bypass (CPB), are associated with the development of cardiac surgery-associated acute kidney injury (CSA-AKI). Development of CSA-AKI occurs as a result of inflammation, uncontrolled complement activation, and kidney cell damage. In patients with preoperative renal impairment, such as those with chronic kidney disease (CKD), there is an increased risk of both CSA-AKI and poorer clinical outcomes. Currently, there are limited effective, targeted pharmacological interventions for the prevention or treatment of CSA-AKI, although emerging therapies are being investigated, particularly in patients with existing CKD. The ARTEMIS (RAvulizumab to PRotect PaTients with Chronic Kidney DisEase froM CSA-AKI and Subsequent Major Adverse Kidney Events) trial will assess the efficacy and safety of ravulizumab (a complement C5 inhibitor) in reducing the risk of major adverse kidney events (MAKE) in patients with preoperative CKD undergoing non-emergent cardiac surgery with CPB.<h4>Methods</h4>This trial is currently recruiting patients with CKD who have planned cardiac surgery requiring CPB including coronary artery bypass grafting, valve replacement or repair, or combined procedures. This is a phase 3, randomized, double-blind, placebo-controlled, global study assessing the efficacy and safety of a single preoperative dose of ravulizumab. These outcomes will be assessed using the occurrence of MAKE and its components, as well as the occurrence and severity of CSA-AKI throughout the study period.<h4>Discussion</h4>Complement activation is known to occur during and after cardiac procedures as a result of CPB and ischemia-reperfusion injury, leading to a cycle of cell damage and death. Therefore, it is hypothesized that preoperative administration of ravulizumab will provide immediate and complete complement inhibition, which will be sustained throughout the surgical period, preventing the uncontrolled complement activation associated with the development of CSA-AKI, thus minimizing poor outcomes for patients.<h4>Trial registration</h4>ClinicalTrials.gov NCT05746559. Registered on February 27, 2023.

<h4>Objective</h4>The presence of overexpressed folate-receptor and high concentration of glutathione (GSH) in liver cancer cells has been exploited and we have synthesized Folic acid-Dextran-Cystamine-Stearic acid (FDCS) for efficient and multifunctional delivery of the drug sorafenib (SAF) to enhance the anticancer effects.<h4>Methods</h4>The characteristics of micelles such as physicochemical properties and in vitro release were investigated. The pharmacokinetic characteristics of the micelle and SAF groups in rats were investigated. In vitro and in vivo anti-tumor experiments were performed with HepG2 cells.<h4>Results</h4>SAF-FDCS was successfully prepared and characterized. The cellular experiments showed that SAF-FDCS significantly enhanced the toxicity and inhibitory effects on HepG2 cells compared with free drug and the other micelles without smart response, and the uptake capacity of cellular HepG2 for SAF-FDCS was significantly higher than the groups without folic acid. Pharmacokinetic results showed that SAF-FDCS revealed a longer circulation time than free SAF. In addition, the tumor inhibition rate of SAF-FDCS in the subcutaneous graft tumor model of HepG2 cells was 84.6%, significantly higher than in other groups.<h4>Conclusions</h4>These results demonstrated the feasibility of SAF-FDCS micelles in inhibiting tumor growth and their superiority in anti-cancer effects compared to free drugs and normal micelles.

Resective surgery is an effective approach for long-term seizure control in drug-resistant focal epilepsy when the epileptic focus (EF) can be accurately delineated and removed. However, intraoperative mapping of EF with electrocorticography is laborious, time-consuming, and highly vulnerable to the effects of anesthesia. Here, we demonstrated that activated microglia can be reliable biomarkers for EF localization. Leveraging a newly developed ratiometric Raman nanosensor, ultraHOCls, we successfully visualize proinflammatory microglia in live epileptic mice, allowing for precise EF delineation without the interference of anesthesia. Compared to electrocorticography-guided surgery, ultraHOCl-guided surgery results in a substantial 61% reduction in total seizure burden in epileptic mouse models. Notably, ultraHOCls sprayed on freshly excised human brain tissues can effectively discriminate epileptic regions from non-epileptic tissues with high sensitivity (94.89%) and specificity (93.3%). This work provides an alternative strategy for delineating the EF intraoperatively, potentially revolutionizing surgery outcomes in epilepsy patients.

T helper 1 (Th1) cell initiation pathways are well characterized; however, those regulating their contraction are less understood. Here, we define a CD4⁺ T cell-autonomous pathway in which complement C5 orchestrated a shift from prostaglandin E2 (PGE2) dominance to enhanced prostacyclin (PGI2) production via activation of C5a receptor 2 (C5aR2). This pivot in lipid mediators induced autocrine signaling through the PGI2 receptor and expression of the interleukin-1 (IL-1) decoy IL-1 receptor type 2 (IL-1R2), which sequestered Th1 cell-driving intrinsic IL-1β, facilitating Th1 cell contraction. Disruption of this C5aR2-PGI2-R axis was a hallmark of pathologically persistent Th1 cell activity in inflammatory conditions, including cryopyrin-associated periodic syndromes (CAPS), Crohn's disease, and rheumatoid arthritis. Rebalancing this axis through selective PGE2 synthase inhibition rectified the hyperactive Th1 cell phenotype in vitro in T cells from individuals with CAPS. Therefore, complement is a key controller of prostanoid metabolism, and the latter is an intrinsic-and potentially druggable-checkpoint for the cessation of Th1 cell effector responses.

Due to exploitation of the mineral resource, the Wumeng Mountain region has become one of the most severely polluted areas. This environmental toxicant accumulations threatens the ecosystem, which is vital for ruminant survival, and poses potential health risks to humans through circulation of contaminated product. The present study examined two pastures. Compared to the CON pasture, sulfur (S) and molybdenum (Mo) concentrations were significantly higher in both the soil and forage of the SMO pasture (p < 0.05). Specifically, the S content in SMO forage reached 28.533 g/kg and the Mo content reached 6.18 mg/kg, both of which far exceeded the recommended levels for small ruminants (1-2 g/kg for S and 0.05-0.1 mg/kg for Mo) as outlined by the NRC (2005). Therefore, the SMO pasture was under environmental S and Mo stress. Forty 4-5-month-old male Guizhou black goats were selected (20 from each pasture) for 150 grazing days. Environmental S and Mo stress in SMO goats led to secondary copper (Cu) deficiency (p < 0.05) and lower FW and ADG (p < 0.05). Hematological analysis revealed anemia, with decreased HGB, HCT, MCV, and MCH and increased RDW-CV and RDW-SD (p < 0.05). Serum analysis revealed liver function impairment, with elevated ALT, AST, TBIL, ALP, and CREA (p < 0.05), and inflammatory responses, with increased levels of IL-1β, IL-6, IFN-γ and TNF-α in the serum and increased liver mRNA expression of these cytokines, as well as decreased levels of IL-10 in the serum and decreased liver mRNA expression of IL-10 (p < 0.05). TMT-based proteomics identified 173 differentially expressed proteins (84 upregulated and 89 downregulated). KEGG analysis revealed disruptions in the complement, coagulation, oxidative phosphorylation, and cytochrome P450 pathways. Potential biomarkers include SERPINC1, F2, APOH, APOB, PLG, and AMBP. In conclusion, environmental S and Mo stress impairs the mineral balance, physiological functions, immune system, and growth of goats.

Pediatric acute lymphoblastic leukemia (ALL) is among the most prevalent hematological malignancies in children. Despite an overall cure rate approaching 90%, a subset of patients still experiences relapse, even with advanced therapeutic interventions. Research into the molecular characteristics and prognostic markers of fusion gene-negative (FG-negative) pediatric ALL remains limited. To address this gap, we performed whole-exome sequencing (WES) and whole-transcriptome sequencing (RNA-seq) on 54 FG-negative ALL cases from our center. Our results indicated that neither specific mutations nor tumor mutational burden significantly influenced relapse risk. Notably, we identified a significant downregulation of CADPS in FG-negative pediatric ALL patients who relapsed. The expression levels and prognostic significance of CADPS were further validated using data from the Therapeutically Applicable Research to Generate Effective Treatments (TARGET) cohort, where lower CADPS expression was associated with reduced event-free survival (EFS) and overall survival (OS) (P < 0.001 for both). Cox regression analyses were subsequently employed to identify OS-related factors and to construct a prognostic prediction model. Notably, this model demonstrated a significant correlation with therapeutic targets. In conclusion, our findings support the potential of CADPS expression as a novel biomarker for prognostic stratification in FG-negative pediatric ALL patients.

<h4>Background</h4>Androgenetic alopecia (AGA) is the most common type of androgen-associated hair loss. Emerging evidence highlights inflammation as a critical mediator in follicular miniaturization and disease progression. This investigation systematically explores inflammatory mechanisms in AGA through comprehensive analysis of hair follicles transcriptional profiles combined with cellular heterogeneity.<h4>Methods</h4>Matched follicular specimens were procured from AGA patients: occipital non-balding units (controls) versus frontal alopecic zones (experimental). Bulk RNA-sequencing was conducted on Norwood-Hamilton grade 3-5 AGA scalp tissues to delineate inflammatory signatures. Subsequent single-nucleus RNA sequencing (snRNA-seq) of grade 5 specimens resolved cellular heterogeneity. Immune subsets (NK/CD8⁺ T cells), vascular endothelia (BECs), keratinocytes, and fibroblasts were transcriptionally characterized. Findings were validated through immunofluorescence cytochemistry (IFC) and reverse transcription quantitative PCR (RT-qPCR).<h4>Results</h4>Bulk RNA-sequencing of AGA hair follicles revealed heightened inflammatory signatures in grade 5 patients compared to grade 3-4 counterparts. To dissect cellular heterogeneity, we systematically investigated the dynamic changes of immune cells in hair follicles of AGA patients using snRNA-seq technology for the first time. The result showed that grade 5 AGA hair follicles, identifying significant enrichment of natural killer (NK) and CD8⁺ T cells in balding hair follicles. Concurrently, blood endothelial cells (BECs) in balding follicles exhibited downregulation of angiogenesis-related genes. Notably, IL-15-a cytokine critical for NK/CD8⁺ T cell proliferation-was overexpressed in BECs, keratinocytes, and fibroblasts, suggesting a microenvironmental cue for immune cell expansion.<h4>Conclusion</h4>These findings collectively implicate NK and CD8⁺ T cell infiltration as drivers of inflammatory exacerbation in AGA. By blocking IL-15 signaling-mediated immune activation may be an innovative therapeutic approach to promote hair regeneration in AGA patients.

Male fertility is strongly influenced by environmental exposures, lifestyle, and advancing age. While advanced paternal age (APA) has been linked with a progressive decline in male fertility, poor reproductive outcomes, and decreased offspring health, the molecular mechanisms underlying these alterations remain unclear. In this work, we investigated the impact of men's age on human sperm protein expression and phosphorylation to identify molecular alterations possibly responsible for the age-associated decline in male fertility. Semen samples from volunteers attending fertility consultations at the Hospital of Aveiro were collected, analyzed according to WHO's guidelines, and processed by the density gradient technique. The proteome and phosphoproteome of 19 normozoospermic human sperm samples divided into four age groups were evaluated by mass spectrometry: ≤30 years old; 31-35 years old; 36-40 years old; and >40 years old. Proteomic analysis revealed 46 differentially expressed proteins (DEPs) between groups, some of them associated with infertility-related phenotypes. Gene ontology (GO) analysis, performed using the DAVID database, revealed that DEPs in older men were enriched in pathways related to stress response, metabolism, and embryo implantation. Additionally, 94 differentially phosphorylated sites corresponding to 76 differentially expressed phosphorylated proteins between the groups were identified, related to key reproductive processes such as sperm motility, spermatogenesis, and sperm binding to zona pellucida, and involved in metabolic and stress response pathways, like HSF1 activation. The set of proteins and phosphorylated residues altered in the sperm fraction usually used in assisted reproductive technology (ART) highlights the need to consider the age of the male partner during fertility assessment and treatment planning. These markers can also be used to explain cases of idiopathic infertility, failure in ART, or repeated abortion associated with APA, overcoming the subjectivity of the conventional semen analysis.

Cataract formation remains a significant cause of global visual impairment. Increasing attention has been directed toward antioxidant-based interventions as potential non-surgical strategies to delay or prevent cataractogenesis, particularly in the age-related and diabetic contexts. This review summarizes recent preclinical evidence on nutritional antioxidants for the prevention of age-related and diabetic cataracts. Agents such as trimetazidine, Moringa oleifera stem extract, ginsenoside Rg1, lanosterol nanoparticles, β-casomorphin-7, and cerium oxide-based nanotherapies have been shown to mitigate oxidative damage, modulate redox signaling pathways, and preserve lens clarity. Advances in drug delivery, including topical formulations, nanoparticle carriers, and intravitreal injections, have been proposed to overcome the anatomical and pharmacokinetic barriers associated with the avascular lens. The new data support ongoing translational research to maximize the clinical use of antioxidants and highlight their therapeutic potential in the prevention of age-related and diabetic cataracts.

<b><b>Background/Objectives:</b></b> Preeclampsia is a multifactorial disorder with a possible genetic component. While numerous studies have explored genetic susceptibility, validation remains inconsistent. The aim was to assess the association between hypertension-related polymorphisms and preeclampsia risk. <b>Methods</b>: A case-control study was conducted in Karaganda, Kazakhstan (n = 95). Sixty SNPs were genotyped using the QuantStudio™ 12K Flex system. Genotype-phenotype associations were evaluated using five inheritance models and statistical analysis in R. <b>Results</b>: Significant associations were found for rs2516839 (C/T: OR = 5.28; 95% CI: 1.53-18.15), rs17672135 (T/T: OR = 3.48; CI: 1.05-11.5), and rs10757278 (A/G: OR = 0.3; CI: 0.11-0.83). However, wide confidence intervals suggest potential limitations in sample size and generalizability. <b>Conclusions</b>: While these polymorphisms show promise as genetic markers of preeclampsia risk, their clinical application requires further validation in larger, multi-ethnic cohorts.

Iron accumulation in the brain is widespread in Alzheimer's disease (AD), the most common cause of dementia. According to numerous studies, too much iron triggers the development of neurofibrillary tangles (NFTs) and amyloid-β (Aβ) plaques, both of which accelerate the onset of AD. Iron sequestration and storage were disrupted by high iron, and the pattern of interaction between iron regulatory proteins (IRPs) and iron-responsive elements (IREs) was altered. The 5'-untranslated regions (5'-UTRs) of their APP mRNA transcripts have an IRE stem-loop, which is where iron influx enhances the translation of the amyloid precursor protein (APP). Iron regulated APP expression via the release of the repressor interaction of APP mRNA with IRP1 by a pathway similar to the iron control translation of the ferritin mRNA by the IREs in their 5'-UTRs. This leads to an uncontrolled buildup of redox active Fe²⁺, which exacerbates neurotoxic oxidative stress and neuronal death. Fe²⁺ overload upregulates the APP expression and increases the cleavage of APP and the accumulation of Aβ in the brain. The level of APP and Aβ, and protein aggregates, can be downregulated by IRPs, but are upregulated in the presence of iron overload. Therefore, the inhibition of the IRE-modulated expression of APP or Fe²⁺ chelation offers therapeutic significance to AD. In this article, I discuss the structural and functional features of IRE in the 5'-UTR of APP mRNA in relation to the cellular Fe²⁺ level, and the link between iron and AD through the amyloid translational mechanism. Although there are currently no treatments for AD, a progressive neurodegenerative disease, there are a number of promising RNA inhibitor and Fe²⁺ chelating agent therapeutic candidates that have been discovered and are being validated in April 2025 clinical trials. Future studies are expected to further show the therapeutic efficacy of iron-chelating medications, which target the APP 5'-UTR and have the ability to lower APP translation and, consequently, Aβ levels. As a result, these molecules have a great deal of promise for the development of small-molecule RNA inhibitors for the treatment of AD.

<h4>Background</h4>The proper development of embryonic stomach in human is essential for the functionality of the adult stomach. However, the key genes, biological processes, and signaling pathways that influence stomach development in human embryogenesis are not yet fully understood.<h4>Methods</h4>In this study, stomach samples were obtained from human embryos at developmental stages ranging from two to seven months. Through transcriptomic sequencing, we identified the differentially expressed genes and enrichment processes in the stomach at various developmental phases.<h4>Results</h4>The results of this study indicate that genes associated with embryonic organ morphogenesis, digestive tract development, and gastric acid secretion displayed elevated expression during the early developmental stages. Additionally, a number of genes linked to cilium assembly and organization, peptide and hormone secretion and transportation, and immune response, showed increased expression during stomach maturation. Our findings elucidate that both the morphological and functional aspects of the stomach develop during the early stages of embryonic development. As gastric development, the stomach progressively acquires additional functions. This research provides insights into the intricate regulatory networks among the genes involved in embryonic digestive tract development, digestion and embryonic organ morphogenesis. Therefore, the formation of human embryonic stomach necessitates the synergistic regulation of a plethora of genes. Notably, this study not only identified traditionally recognized genes but also revealed many previously uncharacterized genes that play potential roles in stomach development and its functions.<h4>Conclusion</h4>These findings establish a crucial basis for future studies on stomach development and the disorders arising from fetal stomach abnormalities.

Adult mammalian brains generate new neurons throughout life in two main niches, the dentate gyrus of the hippocampus and the subventricular zone, starting from neural stem cells (NSCs). Adult hippocampal neurogenesis is crucial for learning and memory and decreases during aging. As defined in mouse models, NSCs, which are prevalently quiescent, develop into proliferating progenitor cells, neuroblasts, and immature and mature neurons. Two visions for NSC self-renewal in the dentate gyrus have been proposed, one postulating persistent self-renewal, with cycles of rest and reactivation even in old age, and the other proposing a short-lived NSC model. Single-cell RNA sequencing and clonal studies, discussed in this review, have shed light on the developmental steps of neurogenic cells and the modality of self-renewal, revealing the presence in the adult dentate gyrus of NSC heterogeneous populations, one long-lived and another rapidly depleted at an early age. Another relevant question is whether adult neurogenesis occurs in humans. A few single-cell RNA-seq studies show that new neurons, with prolonged neuronal maturation, are continuously generated at low frequency from stem/progenitor cells, which results in the accumulation of immature granule cell neurons. This suggests an important role of these cells in human neurogenesis and hence interspecies differences in the neurogenic process dynamics. This review is focused on transcriptomic studies that have faced these and other NSC issues by analyzing developmental trajectories of neural cells and NSCs gene expression profiles in specific experimental settings of hippocampal neurogenesis, and also in mouse models with deletion or overexpression of specific genes to reproduce neural pathologies.

<h4>Background</h4>Dynamic changes in the mammary gland during lactation and the dry period involve proliferation, secretion, apoptosis, and remodeling of mammary epithelial cells. MicroRNAs (miRNAs) are recognized as critical regulators of mammary gland development and lactation. However, their expression patterns and regulatory mechanisms at different lactation stages-particularly during mammary involution and remodeling-remain poorly understood in dairy goats.<h4>Methods</h4>In this study, high-throughput sequencing was employed to analyze miRNA expression profiles in goat mammary tissues at five key stages: late gestation (LG), early lactation (EL), peak lactation (PL), late lactation (LL), and the dry period (DP). Differential expression analysis, miRNA clustering, Gene Ontology (GO) annotation, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment were performed to predict the functions of target genes. A miRNA-mRNA regulatory network associated with mammary gland development was constructed, and functional validation experiments were conducted to confirm key regulatory relationships.<h4>Results</h4>A total of 1,120 miRNAs were identified, including 408 known and 712 newly predicted miRNAs. Among them, 383 were significantly differentially expressed, with the largest number observed between the dry period and late gestation. Six expression-specific miRNA clusters were identified. Functional enrichment analysis indicated that these miRNAs may regulate epithelial cell proliferation, apoptosis, and tissue remodeling by targeting pathways such as energy metabolism, cell adhesion, and the PI3K/Akt signaling pathway. IGF1R was identified as a key regulatory gene in the miRNA-mRNA network related to mammary gland development. Experimental validation showed that chi-miR-423-3p inhibited mammary epithelial cell proliferation, induced G1/S cell cycle arrest, and promoted apoptosis by targeting IGF1R and suppressing the PI3K/Akt pathway.<h4>Conclusion</h4>This study highlights the dynamic regulatory roles of miRNAs in the goat mammary gland across lactation stages. Notably, the miR-423-3p/IGF1R axis is a key regulator of mammary remodeling during the dry period, offering new insights into the molecular basis of mammary gland functional transitions.

In the past few years, UAV application in agriculture has increased significantly due to higher efficiency and safety, simple operation, reduced labor requirements, and saving chemicals as compared to conventional sprayers. The UAVs are widely used in agriculture, providing flexibility and more profit to farmers. In recent years, research has been conducted on various operational parameters of UAV, and there has been no experiment or study on the effect of operational parameters and drift characteristics of UAV and control of thrips in pigeon pea crop. Therefore, this study evaluated the effect of the operational parameters of a UAV-based spraying system on the performance of agrochemical application in pigeon pea crop to control thrips. A field study was conducted to determine the performance parameters in terms of droplet deposition density, droplet size, coverage, spray deposition, and relative span at different flight heights (1.5, 2, and 2.5 m above the crop canopy) and flight speeds (2, 2.5, and 3 m/s). Water-sensitive papers (WSPs) were placed at three canopy zones (bottom, middle, and top zones) of the pigeon pea plant. The maximum droplet density, droplet size, coverage, spray deposition, and relative span factor at the top, middle, and bottom canopy zones were 54.00, 50.17, and 46.33 droplets/cm²; 244.80, 239.88, and 235.37 µm; 10.53%, 10.09%, and 9.78%; 0.764, 0.714, and 0.672 µl/cm²; and 0.98, 0.96, and 0.93, respectively, at a flight height of 1.5 m and a flying speed of 2 m/s. Spray deposition was negligible in off-target zones. The field capacity, field efficiency, and application rate of the UAV were found to be 2.62 ha/h, 60.64%, and 77.86 L/ha, respectively. The maximum control efficacy of thrips on the top, middle, and bottom pigeon pea canopy was 92.45%, 90.12%, and 88.11% after 10 days of spraying experiment. This study provides recommendations for optimal operating parameters (height: 1.5 m and speed: 2 m/s) for efficient agrochemical application, benefiting manufacturers, farmers, and UAV operators for more effective and efficient spraying on pigeon pea crops.

Immune checkpoint inhibitors (ICIs) have revolutionized the treatment for different types of cancers, providing significant clinical benefits. However, these therapies are associated with various immune-related adverse events (irAEs), including hepatic manifestations such as hepatitis, sinusoidal obstruction syndrome (SOS), and nodular regenerative hyperplasia. Among these, regenerative hepatic pseudotumors (RHPs) are exceptionally rare and poorly described in literature. Here, we report the case of a 66-year-old man with metastatic non-small-cell lung cancer (NSCLC) who developed a hepatic pseudotumor during routine imaging following treatment with the anti-programmed cell death 1 (PD-1) therapy, tislelizumab. Despite the presence of a hepatic lesion on imaging, the patient exhibited no clinical symptoms or biochemical evidence of severe immune-mediated hepatitis. Following cessation of anti-PD-1 therapy and initiation of systemic steroid therapy, the hepatic pseudotumors stabilized without further growth. The findings suggest that ICI therapy may be associated with the development of regenerative hepatic pseudotumor (RHP). Given the nonspecific and potentially misleading imaging features of RHP, biopsy is essential for accurate diagnosis and differentiation from malignant lesions such as hepatic metastases. Early histological evaluation through biopsy can prevent unnecessary interventions and guide appropriate management in patients presenting with liver lesions during or after ICI therapy. This case suggests a possible association between the development of RHP and tislelizumab treatment. The effect of ICI-induced hepatic pseudotumors on NSCLC progression is unclear and requires further investigation.

Colorectal cancer is a common malignant tumor in the gastrointestinal tract, and the mechanisms of its occurrence, development, and metastasis have always been the focus of the medical community's attention. The study of CRC genetic mechanisms began with the identification of oncogenes or tumor suppressor genes and their key pathways. With further research, researchers gradually realized that single genes or pathways alone could not explain the occurrence, development, and metastasis of CRC. The development of bulk sequencing technology has helped us to analyze the occurrence, development, and metastasis mechanisms of CRC from a multi-gene, multi-pathway, and multi-dimensional perspective, but it has not brought significant benefits to the clinical treatment of tumors. The main reason for this is that bulk sequencing technology relies on homogeneous cell grouping and cannot capture the heterogeneity between cells within the tumor and the interactions within the tumor microenvironment. The development of single-cell technology has made it possible to study the mechanisms of heterogeneity between cells within CRC and the interaction within the tumor microenvironment. This review discusses the mechanisms of CRC occurrence and development in three stages: traditional molecular biology level of single gene, bulk sequencing, and single-cell sequencing. These results show that the occurrence of CRC is the result of complex interactions between genetic and non-genetic factors in somatic cell evolution, where the heterogeneity between cells within the tumor and the tumor microenvironment are crucial for CRC progression.

Defined as the semi-autonomous orientation to either a moving or rapidly appearing stimulus, reflexive attention (RA) is a crucial process for humans. While there are multiple outcomes used to assess RA, their relationships have not been tested. Disruptions in RA often relate to neurodevelopmental disorders, such as attention-deficit/hyperactivity disorder or autism spectrum disorder, so RA might be moderated by age. Additionally, the current academic consensus is that multiple genes influence RA, but this has not been assessed in a meta-analysis. A better understanding of RA using previously collected data will allow us to improve the design of future research studies.<h4>Methods</h4>In accordance with PRISMA, we conducted a broad search for potentially relevant articles pertaining to genes associated with RA. Selected studies included those (1) published in English, (2) involving human participants, and (3) referencing specific genetic markers in association with a measure of RA. For subgroup comparisons, we analyzed 14 studies assessing children and 23 assessing adults. We also compared 18 dopamine-related to 19 non-dopamine related studies.<h4>Results</h4>The main analysis produced a non-significant overall effect size; however, our most interesting finding was that results varied by age group. We explore this as well as difference by outcome type and the relation of the gene studied to dopamine.<h4>Conclusion</h4>Our findings vary by age group. However, due to heterogeneity we recommend more studies to answer some questions about a broader range of neurotransmitters, to include younger age groups, and to clarify difference by outcome type. We discuss issues of relevance to researchers to guide future meta-analyses.<h4>Systematic review registration</h4>Prospero: International prospective register of systematic reviews. Available from: https://www.crd.york.ac.uk/PROSPERO/view/CRD42018090220.

Transcription factor (TF) proteins play a critical role in the regulation of eukaryote gene expression via sequence-specific binding to genomic locations known as TF binding sites. We studied sites of genomic variation between modern human and Neanderthal promoters. We detected significant differences in the binding affinities of 110 TFs to the promoters of 75 target genes. The TFs were enriched for terms related to vision, motor neurons, homeobox, and brain, whereas the target genes and their direct interactors were enriched in terms related to autism, brain, connective tissue, trachea, prostate, skull morphology, and vision. Secondary analysis of single-cell data revealed that a subset of the identified TFs (CUX1, CUX2, ESRRG, FOXP1, FOXP2, MEF2C, POU6F2, PRRX1 and RORA) co-occur as marker genes in L4 glutamatergic neurons. The majority of these genes have known roles in autism and/or schizophrenia and are associated with human accelerated regions (elevated divergence in humans vs. other primates). Analysis of a single-nucleus dataset of cortical tissue showed that 15 of these TFs and 16 of their target genes are differentially expressed in autism vs. control, most commonly upregulated in developing neurons. Down regulation of these genes occurred in SV2C- and somatostatin-expressing interneurons, oligodendrocytes, and oligodendrocyte precursor cells. These results support the value of gene regulation studies for the evolution of human cognitive abilities and the neuropsychiatric disorders that accompany it.

A protocol for the preparation of fatty acid-functionalized nanohydroxyapatite was developed. These nanoparticles were characterized and used as building blocks for the preparation of Pickering water-in-oil emulsions, which in turn were used as a template for colloidosomes. It is possible to encapsulate proteins in these structures, and their low cellular toxicity was also demonstrated. These structures show structural fragility to pH and temperature changes.

With the gradual advancement of research methods and technologies, various biological processes have been identified as playing roles in the pathogenesis of neurodegenerative diseases. However, current descriptions of these biological processes do not fully explain the onset, progression, and development of these conditions. Therefore, exploration of the pathogenesis of neurodegenerative diseases remains a valuable area of research. This review summarizes the potential common pathogeneses of Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, Huntington's disease, frontotemporal lobar dementia, and Lewy body disease. Research findings have indicated that several common biological processes, including aging, genetic factors, progressive neuronal dysfunction, neuronal death and apoptosis, protein misfolding and aggregation, neuroinflammation, mitochondrial dysfunction, axonal transport defects, and gut microbiota dysbiosis, are involved in the pathogenesis of these six neurodegenerative diseases. Based on current information derived from diverse areas of research, these biological processes may form complex pathogenic networks that lead to distinctive types of neuronal death in neurodegenerative diseases. Furthermore, promoting the regeneration of damaged neurons may be achievable through the repair of affected neural cells if the underlying pathogenesis can be prevented or reversed. Hence, these potential common biological processes may represent only very small, limited elements within numerous intricate pathogenic networks associated with neurodegenerative diseases. In clinical treatment, interfering with any single biological process has proven insufficient to completely halt the progression of neurodegenerative diseases. Therefore, future research on the pathogenesis of neurodegenerative diseases should focus on uncovering the complex pathogenic networks, rather than isolating individual biological processes. Based on this, therapies that aim to block or reverse various targets involved in the potential pathogenic mechanisms of neurodegenerative diseases may be promising directions, as current treatment methods that focus on halting a single pathogenic factor have not achieved satisfactory efficacy.

Colorectal cancer (CRC) remains a leading cause of cancer-related mortality, driven by complex interactions between inflammatory pathways, gut microbiota dysbiosis, and tumor microenvironment remodeling. Conventional therapies, particularly single-target oral chemotherapeutics, are hindered by poor bioavailability, systemic toxicity, and drug resistance. To address these limitations, we engineered KGM-PTX/CSM microspheres, a dual-responsive drug delivery system leveraging the elevated reactive oxygen species (ROS) in CRC and β-mannanase overexpression in the colorectum. The system comprises ROS-sensitive prodrug micelles (PSM) encapsulated within konjac glucomannan (KGM). PSM micelles were synthesized by conjugating hydrophilic chitosan oligosaccharides (COS) with the hydrophobic anti-inflammatory agent mesalazine (MSL) via ROS-labile thioether bonds, followed by paclitaxel (PTX) encapsulation. Upon oral administration, KGM undergoes β-mannanase-triggered degradation in the colon, releasing PSM micelles that subsequently disintegrate in the ROS-rich tumor microenvironment, enabling spatiotemporally controlled drug release. <i>In vitro</i> studies demonstrated ROS-responsive drug liberation (91.2 % cumulative release within 48 h) and enhanced cytotoxicity against PTX-resistant SW480/PTX cells (IC₅₀: 9.33 μg/mL vs. 45.68 μg/mL for free PTX). Mechanistic investigations revealed synergistic interactions among the system's components: PTX stabilized microtubules to induce apoptosis, while MSL counteracted COX-2/P-gp-mediated drug resistance and alleviated PTX-associated intestinal inflammation. In the AOM/DSS-induced orthotopic CRC model, KGM-PTX/CSM significantly inhibited colorectal tumor growth, improved survival rates, and suppressed inflammatory cytokine expression (TNF-α, IL-1β, IL-6, and IL-10) in serum and colorectal tissues. Immunomodulatory effects included enhanced CD8⁺ T-cell activity, suppression of Treg-mediated immune evasion, and macrophage polarization toward the tumor-suppressive M1 phenotype. Gut microbiota analysis demonstrated restored operational taxonomic unit (OTU) counts, increased beneficial bacterial populations, elevated alpha and beta diversity, reduced pro-inflammatory bacteria, and increased short-chain fatty acid (acetate, propionate, and butyrate) concentrations, collectively improving intestinal microecology and inhibiting tumor progression. This study synergistically enhanced the anti-CRC effect through multiple mechanisms of action such as chemotherapy, reversal of chemotherapy resistance, regulation of intestinal flora, anti-inflammation, activation of immune cells, etc., which will provide a certain reference for the research of synergistic drug therapy for CRC.

Allogeneic hematopoietic cell transplantation (alloHCT) is the sole curative therapy for myelodysplastic syndrome (MDS). While alloHCT clearly confers a significant survival advantage in high-risk MDS, it is less clear how the disease burden and impact of conditioning intensity impact survival. This review addresses critical issues surrounding this topic, emphasizing the unique cell biology of MDS and the evolving concepts of conditioning intensity compared to other diseases, including acute myeloid leukemia (AML). The review is structured around three interconnected themes. First, it clarifies the varying interpretations of conditioning intensity. Second, it examines the interplay between disease burden at transplant and conditioning intensity in determining outcomes, including a comparative analysis with acute myeloid leukemia (AML) to highlight similarities and differences. Third, it explores the relationship between conditioning regimen intensity and immune reconstitution, particularly focusing on the graft-versus-tumor (GvT) effect and its potential modulation by conditioning intensity. Understanding the stem cell target of conditioning regimens is emphasized, as the persistence of the underlying MDS stem cell necessitates a thorough understanding of this concept for improved therapeutic strategies.

<b>Objectives:</b> This study aimed to compare in vitro the protective effect of two enamel remineralizing agents, a varnish containing β-tricalcium phosphate with sodium fluoride (β-TCP-F) and a paste containing casein phosphopeptide-amorphous calcium phosphate with sodium fluoride (CPP-ACP-F), on artificially demineralized human enamel. <b>Methods:</b> A total of 120 human third molar enamel specimens were randomly assigned to four groups (n = 30 each): Group I (healthy enamel, control), Group II (initially demineralized, lesioned enamel), Group III (demineralized enamel and treated with β-TCP-F), and Group IV (demineralized enamel and treated with CPP-ACP-F). Groups II-IV underwent, for 15 days, a daily pH cycling regimen consisting of 21 h of demineralization under pH 4.4, followed by 3 h of remineralization under pH 7. Groups III and IV were treated with either β-TCP-F or CPP-ACP-F, prior to each 24 h demineralization-remineralization cycle. Fluoride ion release was measured after each pH cycle. Surface hardness, roughness, wettability, and <i>Streptococcus mutans</i> biofilm formation were assessed on days 5, 10, and 15 after a daily pH cycle. <b>Results:</b> CPP-ACP-F treatment showed a larger improvement in surface hardness (515.2 ± 10.7) compared to β-TCP-F (473.6 ± 12.8). Surface roughness decreased for both treatments compared to initially lesioned enamel; however, the decrease in roughness in the β-TCP-F group only reached a value of 1.193 μm after 15 days of treatment, a significantly larger value in comparison to healthy enamel. On the other hand, the decrease in roughness in the CPP-ACP-F treatment group reached a value of 0.76 μm, similar to that of healthy enamel. Contact angle measurements indicated that wettability increased in both treatment groups (β-TCP-F: 71.01°, CPP-ACP-F: 65.24°) compared to initially lesioned samples in Group II, reaching WCA values similar to or smaller than those of healthy enamel surfaces. <b>Conclusions</b>: Both treatments, β-TCP-F and CPP-ACP-F, demonstrated protective effects against enamel demineralization, with CPP-ACP-F showing superior enhancement of surface hardness and smoother enamel texture under in vitro pH cycling conditions. β-TCP-F varnish and CPP-ACP-F paste treatments counteracted surface modifications produced on human healthy enamel by in vitro demineralization.

<b>Background/Objectives:</b> Cancer and bacterial cases are increasing. Hence, new drugs to treat these diseases are paramount. Ferrocene-based hybrid compounds were synthesizedas potential cancer and bacteria therapeutics. <b>Methods:</b> The synthesized compounds were characterized via FTIR, NMR, and LC-MS and evaluated against different cancer cells and bacterial strains. Moreover, computational studies of these compounds were conducted using several silico tools. <b>Results:</b> Among the synthesized compounds, hybrid <b>10</b> was the most promising compound, displaying promising anticancer activity with IC₅₀ values between 42.42 and 45.37 and 50.64 and 73.37 µg/mL against HeLa and CHO cancer cells, respectively, with a selective index greater than one on HeLa cancer cells. Compounds <b>22</b>-<b>26</b> displayed promising antibacterial activity with a MIC value of 7.8125 µg/mL against most bacterial strains in vitro. The in silico results revealed that this compound has strong binding affinities for 4qtb, 3eqm, and 2w3l cervical cancer proteins, exhibiting binding energies of -7.3, -8.7, and 7.4 kcal/mol, respectively. Furthermore, hybrid <b>10</b> showed promising pharmacokinetics and drug-like properties, including high GI absorption, moderate water solubility, favoring the oral administration route, nontoxicity, and is a P-gp substrate. <b>Conclusions:</b> The findings obtained in this study illustrate that hybrid compounds are potential therapeutics that need to be explored. The compounds also contained functionalities relevant for incorporating into nanocarriers to improve their biological activities further. Therefore, further studies are recommended for the most effective compounds to reinforce these findings.

<b>Background:</b> Huntington's Disease (HD) is a neurodegenerative disorder caused by an abnormal extension of a CAG repeat stretch located in the exon 1 of the <i>HTT</i> (IT15) gene, leading to production of a mutated and misfolded Huntingtin protein (muHTT) with an abnormally elongated polyglutamine (polyQ) region. This mutation causes muHTT to oligomerize and aggregate in the brain, particularly in the striatum and cortex, causing alterations in intracellular trafficking, caspase activation, and ganglioside metabolism, ultimately leading to neuronal damage and death and causing signs and symptoms such as chorea and cognitive dysfunction. Currently, there is no available cure for HD patients; hence, there is a strong need to look for effective therapies. <b>Methods:</b> This study aims to investigate the efficacy of siRNA-containing nano-engineered lipopolymers in selectively silencing the <i>HTT</i> expression in a neuronal model expressing a chimeric protein formed by the human mutated exon 1 of the <i>HTT</i> gene, tagged with GFP. Toxicity of lipopolymers was assessed using MTT assay, while efficacy of silencing was monitored using qRT-PCR, as well as Western blotting/flow cytometry. Changes in muHTT-GFP aggregation were observed using fluorescence microscopy and image analyses. <b>Results:</b> Here, we show that engineered lipopolymers can be used as delivery vehicles for specific siRNAs, decreasing the transcription of the mutated gene, as well as the muHTT protein production and aggregation, with Leu-Fect C being the most effective candidate amongst the assessed lipopolymers. <b>Conclusions:</b> Our findings have profound implications for genetic disorder therapies, highlighting the potential of nano-engineered materials for silencing mutant genes and facilitating molecular transfection across cellular barriers. This successful in vitro study paves the way for future in vivo investigations with preclinical models, offering hope for previously considered incurable diseases such as HD.

The residues of fluoroquinolones in foods of animal origin posed many threats to the consumer's health. The aim of this study was to developed a magnetic molecularly imprinted microsphere and aggregation induced emission luminogen based semi-homogeneous method for determination of fluoroquinolones in milk. Results showed the limits of detection for determination of 20 fluoroquinolones in milk were in the range of 0.4-11 pg/mL. The sensitivities were increased for up to 4329 folds in comparison with fluorescein isothiocyanate labeled tracer. Due to the ultra-high sensitivity, the milk sample could be analyzed directly after a 100-fold dilution, and the recoveries from fluoroquinolones fortified blank milk samples were in the range of 69.68 %-97.19 %. The method performances were better than the previous fluorescent (immune)assays for fluoroquinolones. This is the first paper reporting the use of magnetic molecularly imprinted microsphere and aggregation induced emission luminogen for determination of veterinary drug in food sample.

Introduction Metabolic dysfunction-associated fatty liver disease (MAFLD) has become the most prevalent chronic liver disease worldwide, particularly in South Asian countries like Pakistan. Gallstone disease (GSD), often associated with metabolic abnormalities, shares several risk factors with MAFLD, such as obesity, diabetes, and dyslipidemia. Despite international evidence supporting an association between MAFLD and GSD, there is a hdlscarcity of local data in this regard. Therefore, this study aimed to determine the independent predictors of GSD in MAFLD patients in Pakistan. Study methodology This cross-sectional study was conducted at the Hepatogastroenterology Department of Sindh Institute of Urology and Transplantation (SIUT) from January to December 2024. A total of 350 adults with the presence of fatty liver on ultrasound were enrolled. Patients with significant alcohol intake, those with known liver diseases, prior cholecystectomy, pregnancy, and use of lipid-altering medications were excluded from the study. Demographic, clinical, and laboratory parameters were recorded. Gallbladder stones were identified using abdominal ultrasound. Univariate analysis followed by multivariate logistic regression was performed to identify the independent predictors of GSD. Results Among 350 patients with MAFLD, 102 (29.1%) had gallstones. Patients with gallstones were significantly older (mean age 52.1 vs. 47.1 years, p<0.001), with a higher prevalence of female gender (63.7% vs. 35.4%, p<0.001) and diabetes mellitus (68.6% vs. 48.3%, p = 0.002). Laboratory markers including alkaline phosphatase (ALP), gamma-glutamyl transferase (GGT), total cholesterol, triglycerides, and fasting blood sugar (FBS) were significantly elevated in the gallstone group, whereas high-density lipoprotein (HDL) was significantly lower. Multivariate logistic regression identified increasing age (OR 1.04), female gender (OR 2.73), diabetes mellitus (OR 1.89), elevated ALP (OR 1.02), low HDL (OR 1.76), and elevated FBS (OR 1.31) as independent predictors of gallstones in MAFLD. Conclusion In this study, 29.1% of MAFLD patients had GSD. Older age, female gender, diabetes, elevated ALP, low HDL, and high FBS were the factors independently associated with the presence of GSD in MAFLD. Further multi-centered prospective studies incorporating dietary, hormonal, and genetic factors are required for the validation of these results.

<h4>Backgruound</h4>Type 2 diabetes mellitus (T2DM) is a significant risk factor for hepatocellular carcinoma (HCC) in patients with nonalcoholic fatty liver disease; however, surveillance strategies for patients with T2DM, especially without cirrhosis, are inadequate. This study examined whether the fatty liver index (FLI) and its dynamic changes can effectively identify patients with T2DM at increased risk for HCC.<h4>Methods</h4>Data from 92,761 individuals with T2DM aged 40 to 79 who underwent two health screenings (2012 to 2015) were analyzed. The FLI, calculated using waist circumference, body mass index, triglycerides, and gamma-glutamyl transferase, was used to stratify patients by baseline FLI and FLI changes between screenings. HCC cases were identified via International Classification of Diseases codes and reimbursement records (2016 to 2020).<h4>Results</h4>Patients with baseline FLI of 30 to 59.9 had a 1.90-fold higher risk (P<0.01) and those with FLI ≥60 had a 2.94-fold higher risk (P<0.01) of developing HCC compared to those with FLI <30. An increase in FLI from <30 to ≥30 resulted in a 2.10-fold higher risk of HCC (P<0.01), while a reduction in FLI from ≥30 to <30 led to a 0.64-fold lower risk (P=0.03). Protective benefits of FLI reduction took approximately 3 years to manifest.<h4>Conclusion</h4>Baseline and dynamic monitoring of FLI effectively identified HCC risk in T2DM patients with non-cirrhotic livers, supporting early detection and intervention.

<h4>Introduction</h4>Lung adenocarcinoma (LUAD) is the most prevalent form of lung cancer worldwide. Long non-coding RNA (lncRNAs) are non-protein coding RNAs that are involved in lung cancer. This study aimed to develop a lncRNA-based risk assessment model based on RNA methylation to evaluate the prognosis of patients with LUAD.<h4>Method</h4>The TCGA-LUAD dataset consisted of 524 primary tumor samples and 59 normal samples, and the validation set (GSE3121011), which included 246 patients with LUAD, was used for this analysis.Pearson's correlation analysis was used to identify lncRNAs associated with RNA methylation in LUAD. Univariate, least absolute shrinkage and selection operators, and multivariate Cox analyses were used to construct the prognostic model. Gene oncology (GO) functional annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were used to identify enriched biological processes. Gene set enrichment analysis (GSEA) and gene set variation analysis (GSVA) were used to visualize the dataset and gene set variation analyses. Kaplan-Meier and decision curve analyses were used to assess the accuracy. qRT-PCR was used to verify the expression of lncRNAs. Invasion and cell scratch assays were conducted to evaluate migration capacity, and colony formation experiments were performed to assess proliferation ability.<h4>Result</h4>Ten RNA methylation-associated lncRNAs were identified to construct risk features. According to the risk model, the patients were categorized into low- and high-risk groups, with the latter exhibiting a less favorable prognosis. The expression levels of the lncRNAs exceeded those in lung epithelial cells. After siRNA transfection, the proliferation and migration abilities of the tumor cells were significantly reduced. The risk-scoring model may be a potential indicator for predicting the sensitivity of patients with LUAD to immunotherapy.<h4>Conclusion</h4>The model constructed in this study can accurately predict the prognosis of patients with LUAD, and holds promise for future immunotherapies.

Vascular smooth muscle cells (VSMCs) exhibit substantial heterogeneity and plasticity, enabling them to switch between contractile and synthetic states, which is crucial for vascular remodeling. Nexilin (NEXN) has been identified as a high-confidence gene associated with dilated cardiomyopathy. Existing evidence indicates NEXN is involved in phenotypic switching of VSMCs. However, a comprehensive understanding of the cell-specific roles and precise mechanisms of NEXN in vascular remodeling remains elusive. Using integrative transcriptomics analysis and smooth muscle-specific lineage-tracing mice, we demonstrated NEXN was highly expressed in VSMCs, and the expression of NEXN was significantly reduced during the phenotypic transformation of VSMCs and intimal hyperplasia induced by vascular injury. VSMC-specific NEXN deficiency promoted the phenotypic transition of VSMCs and exacerbated neointimal hyperplasia in mice following vascular injury. Mechanistically, we found NEXN primarily mediated VSMC proliferation and phenotypic transition through endoplasmic reticulum (ER) stress and Krüppel-like factor 4 signaling. Inhibiting ER stress ameliorated VSMC phenotypic transition by reducing cell cycle activity and proliferation caused by NEXN deficiency. These findings indicate targeting NEXN could be explored as a promising therapeutic approach for proliferative arterial diseases.

Small extracellular vesicles (small EV) are a conserved means of communication across the domains of life and lately gained more interest in mammalian non-cancerous work as non-cellular, biological therapeutic with encouraging results in recent studies of chronic degenerative diseases. The nucleus pulposus (NP) is the avascular and aneural center of an intervertebral disc (IVD), home to unique niche conditions and affected in IVD degeneration. We investigated autologous and mesenchymal stem cell (MSC) small EVs for their potential to contribute to cell and tissue homeostasis in the NP niche via mass spectrometric proteome and functional enrichment analysis using adult and fetal donors. We compared these findings to published small EV databases and MSC small EV data. We propose several mechanisms associated with NP small EVs: Membrane receptor trafficking to modify signal responses promoting niche homeostasis; Redox and energy homeostasis via metabolic enzymes delivery; Cell homeostasis via proteasome delivery and immunomodulation beyond an association with a serum protein corona. The proteome signature of small EVs generated by NP parent cells is similar to previously published small EV data, yet with a focus on supplementing anaerobic metabolism and redox balance while contributing to the maintenance of an aneural and avascular microniche.

<h4>Purpose</h4>Chromosomal rearrangements drive tumorigenesis through fusion transcripts. Clinical whole-transcriptome sequencing (WTS) enables comprehensive fusion detection, overcoming limitations of targeted methods, but its clinical application has been hindered by technical and cost barriers.<h4>Methods</h4>We validated a clinical bait-capture WTS assay using 78 solid tumor samples across diverse tissue types, including 59 with known fusions or oncogenic splice variants. Sensitivity was assessed against clinically reported fusions from targeted next-generation sequencing or fluorescence in situ hybridization. Analytical performance was evaluated for sensitivity, specificity, limit of detection, and reproducibility across varying RNA inputs and sequencing conditions. Clinical implementation data were also analyzed.<h4>Results</h4>WTS demonstrated a sensitivity of 97.1% and a specificity of 100% for detecting clinically relevant fusions. Detection was robust across tumor types, with RNA input as low as 1 ng. Optimized sequencing thresholds of 60 million reads and 130-bp read length improved performance. The assay was highly reproducible and resistant to common contaminants. Over 6 months of clinical use (410 specimens), WTS identified 29 fusions (7% detection rate), including six clinically relevant fusions undetectable by our institution's previous targeted panel, increasing the fusion yield by 26%.<h4>Conclusion</h4>WTS provides high sensitivity, specificity, and expanded fusion detection, supporting its routine clinical adoption. Future work will explore its broader role in integrated genomic profiling and therapeutic guidance.

<h4>Background and objectives</h4>Variants in the homeostatic iron regulator (<i>HFE</i>) gene are prevalent among individuals of European ancestry and have been linked to an increased risk of dementia. This study aimed to evaluate the effects of <i>HFE</i> p.Cys282Tyr and p.His63Asp variants on serum ferritin levels and the incidence of dementia in a cohort of initially healthy older adults.<h4>Method</h4>This prospective longitudinal study used data from the Aspirin in Reducing Events in the Elderly trial. Participants had no history of cardiovascular disease, dementia, or cognitive decline at enrollment. Genotyping for <i>HFE</i> p.Cys282Tyr and p.His63Asp variants was conducted using microarrays, and baseline serum ferritin concentrations were measured in peripheral blood samples. Dementia diagnoses were confirmed by an adjudication committee over a median follow-up of 6.4 years. Associations were evaluated using Cox proportional hazards models adjusted for related covariates.<h4>Results</h4>The study included 12,174 unrelated, healthy participants of European ancestry aged 70 years or older, comprising 5,583 men (45.9%) and 6,591 women (54.1%). The median age was 73.7 years (interquartile range [IQR]: 71.6-76.9) for men and 73.9 years (IQR: 71.7-77.5) for women. Compared with the wild-type group, men with p.Cys282Tyr+/+ (<i>p</i> = 0.048) and p.Cys282Tyr+/p.His63Asp + genotypes (<i>p</i> < 0.001) had significantly higher baseline ferritin levels. Women with p.His63Asp+/+ (<i>p</i> = 0.015) and p.Cys282Tyr+/p.His63Asp+ (<i>p</i> < 0.001) genotypes also exhibited elevated ferritin levels. No significant association was observed between baseline serum ferritin levels and dementia risk. However, men with p.His63Asp+/+ genotype had a significantly higher risk of incident dementia (adjusted hazard ratio = 2.39, 95% CI 1.25-4.57, <i>p</i> = 0.009) compared with those without <i>HFE</i> variations. This association was not observed in women.<h4>Discussion</h4>Among initially healthy older adults, <i>HFE</i> p.His63Asp homozygosity was associated with a higher risk of incident dementia in men but not women. These findings highlight a potential sex-specific genetic risk factor for dementia and warrant further research into the underlying mechanisms linking p.His63Asp and dementia.

<h4>Background</h4>Recent research underscores a potential correlation between chronic obstructive pulmonary disease (COPD) and frailty, suggesting a shared genetic foundation. However, specific genetic factors and mechanisms underlying this association remain unclear. This study aimed to explore genetic connections between COPD and frailty using genome-wide association studies to enhance our understanding and improve clinical management and prevention strategies for these conditions.<h4>Method</h4>We utilised summary statistics for genome-wide association studies to examine the genetic correlations between COPD and frailty using linkage disequilibrium score regression. Local genetic correlations were evaluated using the ρ-heritability estimates from summary statistics method. Using the established two-sample Mendelian randomization approach, causal relationships have been identified. Shared genetic variants were quantified using a bivariate causal mixture model. Shared loci and single nucleotide polymorphisms were identified by conjoint false discovery rate (conjFDR). Gene enrichment and transcriptome-wide association studies (TWAS) were conducted to explore potential transcriptomic associations across tissues.<h4>Results</h4>We observed a significant genetic correlation between COPD and frailty (Rg =  0.4324, P =  6.09 ×  10 - 26). MiXeR estimated 3,200-shared causal variants. Additionally, we discovered 16 shared loci linked to 91 genes, offering novel insights into gene expression across diverse tissues. The TWAS revealed 25 shared genes, representing a significant advance in understanding the genetic overlap between COPD and frailty. Furthermore, out of the 25 SNPs identified through TWAS, 4 overlapped with the lead SNPs, specifically [HLA-DRB1, PBX3, SLC22A5/OCTN2, SLMAP].<h4>Conclusions</h4>Our study shows a common genetic foundation for COPD and frailty, identifying multiple shared loci and offering insights into their underlying causal connections. These findings enhance our understanding of the biological mechanisms linking these conditions and may guide future research and treatment strategies for related diseases.

Despite extensive research highlighting the pivotal role of NOX4 in the development of various malignancies, no systematic pan-cancer analysis has been conducted to evaluate its comprehensive prognostic value and potential immunological functions. In this study, we explored the prognostic significance of NOX4 and its role in immune modulation across different cancer types. We performed a pan-cancer analysis of NOX4 expression and its prognostic implications using multiple databases, including TCGA, GEPIA, GTEx, UALCAN, TISCH, GDSC, PDB, TIMER, and cBioPortal. This analysis provided a detailed assessment of NOX4 expression profiles, clinical correlations, genetic variants, tumor microenvironment interactions, immune relevance, therapeutic potential, and related gene functions through various multi-omics approaches. To further investigate these findings, we collected clinical samples from selected cancer types and validated NOX4 expression through immunohistochemistry, H&E staining, and RT-PCR. Our results revealed that NOX4 was overexpressed in most tumors, although its expression was significantly reduced in certain renal cancers. Moreover, we observed distinct correlations between NOX4 expression and patient prognosis. Notably, NOX4 expression was significantly associated with tumor infiltration, indicating its potential as a target for immunotherapy. Additionally, NOX4 expression showed significant correlations with immune checkpoint proteins (ICP), tumor mutation burden (TMB), microsatellite instability (MSI), RNA stemness scores (RNAss), DNA stemness scores (DNAss), RNA methylation, and DNA methylation.

A variety of techniques for DNA sequencing, such as specific gene sequencing, whole genome sequencing, or exome sequencing, are currently used to detect single nucleotide variations (SNVs). Although RNA-seq can be used to identify SNVs, studies that employ this approach are uncommon, and those that do often rely on outdated mapping methods or methods that are more suitable for genomic and exomic alignment. In this work, our aim is to apply modern RNA-seq specific alignment method in order to identify SNV in a cohort of HCMP patients, and characterize those SNV to gain insight into possible mechanisms of HCMP pathogenesis. The algorithm of identification of SNV based on transcriptomic sequencing data has been developed and evaluated. The algorithm was evaluated and the optimal quality threshold was determined based on allelic discrimination for the rs397516037 mutation (MYBPC3 c.3697 C > T) among patients. A total of 42,809 SNVs with a quality of 75 or higher were identified in 48 transcriptomes of hypertrophic cardiomyopathy (HCMP) myocardial tissue. Verification of missense and nonsense variants in key HCMP genes using Sanger sequencing confirmed the accuracy of the pipeline results. To identify variants potentially associated with HCMP pathogenesis, a filtration process was conducted based on minor allele frequency, substitution prediction score and ClinVar outcome. 214 missense mutations and 6 nonsense mutations were selected. Together with nonsense mutations, 19 mutations meeting the strictest SIFT and PolypPhen criteria were identified as potential factors influencing HCMP pathogenesis. We have developed and validated a method for identifying SNVs based on transcriptomic data, which can be used to identify putative pathogenic variants. We identified mutations in key HCMP genes MYBPC3 and MYH7 in a cohort of patients. We also found potentially pathologic mutations in genes ANXA6 and FEM1 A and obtained data supporting the role of NEBL in myocardial diseases. This method would be useful in analyzing transcriptomic data available in the Gene Expression Omnibus, but should be used with caution as we have tested it on a specific disease.

This study investigates the effect of amine functional groups in ZIF-8 metal-organic frameworks on the loading and release of 5-fluorouracil (5-FU). The facile and cost-effective solvent-assisted linker exchange (SALE) method was used to exchange 2-methylimidazole (2-MIM) linkers with 3-amino-1,2,4-triazole (Atz) in the ZIF-8 structure, which resulted in a synthesis of ZIF-8A with 22, 53, and 74% Atz exchange, respectively. The prepared nanoparticles were characterized by ¹H-NMR, XRD, FT-IR, FE-SEM, UV-Vis spectroscopy, and zeta potential analysis. Drug encapsulation efficiency results showed 12% for 5-FU@ZIF-8 which increased to 48% for 5-FU@ZIF-8A(53%). Also, the results of in-vitro experiments exhibited the pH-responsive behavior of nanocarriers and slower release for 5-FU@ZIF-8A(53%) compared to 5-FU@ZIF-8. The increase in drug encapsulation efficiency and slower release is due to the presence of the amine functional group in the structure, which improves the host-guest interactions between drug molecules and linkers. Moreover, the MTT assay was performed on MCF-7 and HFF-2 cell lines which revealed that 5-FU@ZIF-8A(53%) exhibited more significant cytotoxicity toward cancer cells while less toxicity toward normal cells compared to 5-FU@ZIF-8. These findings highlight the capability of amine-functionalized ZIF-8 as an effective drug delivery system for 5-FU and demonstrate the potential of the facial and low-cost SALE approach as a promising technique in nanocarrier development.

It is an important cause of death in old age to rupture an abdominal aortic aneurysm. The pathogenesis of AAA has not been fully elucidated, and m⁶A RNA methylation regulators have never been implicated in AAA development. This study aimed to explore the expression profile, potential functions and regulated mechanism of m⁶A RNA methylation in the abdominal aortic aneurysm mice model. A successful AAA mouse model was established using Ang II. M⁶A- methylated RNA Immunoprecipitation (MeRIP) sequencing and RNA sequencing were performed to identify the m⁶A sites in the abdominal aorta walls samples. The expression of m⁶A methylation regulators was analyzed in the datasets and MeRIP-qPCR was performed to verify the results of MeRIP-sequencing. Bioinformatics analysis was used to evaluate the m⁶A patterns and indicate the potential signaling pathway. There were 2039 differentially methylated m⁶A peaks involving 1865 mRNAs in the AAA group relative to the control, of which 1610 peaks in 1466 mRNAs were hypermethylated, and 429 peaks in 410 mRNAs were hypomethylated. The hypermethylated mRNAs in AAA group were primarily enriched in transcription regulation and intercellular signaling, especially the Wnt signaling-associated processes. Hypomethylated m⁶A sites were mainly enriched in G protein-coupled receptor activity and ion channel activity. MeRIP-qPCR suggested that the sequencing data were reliable and accurate. The mRNA expression of 24 m⁶A regulators showed no obvious difference between AAA and the control group, but the m6A methylation levels of three components of methyltransferases complex and one 'readers' were significantly increased. Our study suggested an original viewpoint that the m⁶A modification might be regulated by several unidentified regulation modes or genes in the Ang II-induced AAA mice model, and be closely relevant to the combined effect of m⁶A methylation modification in the Wnt pathway, G protein-coupled receptor, and ion channel-associated genes, which were worthy of further investigation.

Glioblastoma (GBM) lethality stems from uncontrolled growth and infiltration. Using an immunocompetent murine model, we mapped GBM invasion and tumor-associated microglia and macrophage (TAM) interactions. We show that microglia are mobilized ahead of invasion, transforming morphologically and functionally-first forming glial nets around tumor infiltrates and then organizing into 'oncostreams' guiding collective migration. Single-cell RNA sequencing revealed three distinct states for tumor cells and microglia, corresponding to invasive niches versus tumor bulk. The invasive patterns and niche-specific gene signatures of tumor cells and TAMs were validated in human GBMs. We further identified a critical role of plexin-B2 in TAMs for resolving cell collision, aligning GBM cells and restructuring the extracellular matrix. Plexin-B2 ablation in TAMs disrupted invasion tracks, shifting GBM growth from infiltrative to bulk expansion. Understanding niche-specific TAM mobilization and anatomical-functional invasion units opens new strategies to target GBM invasion.

The contribution of non-cancer tumoral microenvironment to cachexia is vastly unclear. Despite advances in understanding the signals involved in cancer cachexia progression, the exact time point of cachexia onset remains unpredictable. The transgenic Kras^LSL-G12D/+;Trp53^flox/flox;Pdx1-Cre (KP₂C) GEMM is a clinically relevant model, with the timing of cancer cachexia progression from the pre-cachectic, early-onset, to severe cachexia showed that the onset of cachexia was associated with differences in muscle wasting. The exact cell-of-origin in different types of non-cancer cells in the tumoral microenvironment and the circulating blood, which drives cachexia, remains unclear. Production of potent pro-cachectic substances that induce skeletal muscle wasting also requires mechanistic analysis. This study analyzed the PBMC and the mouse-derived syngeneic transplants (MDSTs) of KP₂C GEMM in recipient mice and pinpoints the cell-type changes with the timing of cachexia (>10% weight loss) by conducting single-cell expression analysis of cell-type-specific gene expression determinants of cachexia. Single-cell RNA sequencing analysis identified signals in high-quality, specific cell types of PBMC (29,615 cells) and MDST (23,151 cells). The scRNA-seq data identified differentially expressed chitinase 3 like 1 (CHI3L1 encoded by mouse Chi3l1) and chitinase-like 3 (CHI3L3, encoded by Chil3) and that macrophages are significant mediators of early-onset muscle wasting in tumor-bearing mice. C2C12 myoblasts treated with the CHI3L1 recombinant protein suppressed myotube formation and upregulated mRNA expression of Hdac3, Tlr9, Irf3, Tbk1, and Nfkb1. Skeletal muscle-specific conditional Hdac3 knockout in tumor-bearing mice decreased muscle wasting via CHI3L1-HDAC3 signaling. An anti-CHI3L1 monoclonal antibody was administered to target these macrophage populations, and the treatment resulted in suppressed tumor growth, metastatic progression, and protected body weight. Our results support the role of pancreatic tumor-associated macrophages in mediating skeletal muscle wasting and provide a clinically relevant mechanism of progression from the pre-cachectic state to the cachexia onset.

The manchette is a transient skirt-like structure consisting of microtubules (MTs) and filamentous actin (F-actin) surrounding the elongating sperm head during spermiogenesis. It is pivotal in sperm head shaping controlled by the acrosome-acroplaxome-manchette complex, acrosome formation, and flagellar assembly by microtubular-based protein delivery. Defects in the manchette frequently lead to teratozoospermia concomitant with oligozoospermia and asthenozoospermia, but the pathogenic mechanism underlying manchette function and its role in male infertility remain poorly understood. In this review, we systematically described the assembly and disassembly of the manchette, intra-manchette transport (IMT) and its regulatory model, the function and mechanism of manchette and IMT in regulating sperm head shaping and flagellar assembly during spermatogenesis; summarized the research progress of manchette-related genes related to male infertility; and listed the manchette-related proteins in knockout mouse models and clinical cases, which provide the theoretical basis for an in-depth understanding of the molecular mechanism of manchette involved in spermatogenesis and male fertility for understanding the potentially developing treatments for infertility and reproductive disorders.

Paediatric bone sarcomas (e.g. Ewing sarcoma, osteosarcoma) comprise significant biological and clinical heterogeneity. This extreme heterogeneity affects response to systemic therapy, facilitates inherent and acquired drug resistance and possibly underpins the origins of metastatic disease, a key component implicit in cancer related death. Across all cancers, metastatic models have offered competing accounts on when dissemination occurs, either early or late during tumorigenesis, whether metastases at different foci arise independently and directly from the primary tumour or give rise to each other, i.e. metastases-to-metastases dissemination, and whether cell exchange occurs between synchronously growing lesions. Although it is probable that all the above mechanisms can lead to metastatic disease, clinical observations indicate that distinct modes of metastasis might predominate in different cancers. Around 70% of patients with bone sarcoma experience metastasis during their disease course but the fundamental molecular and cell mechanisms underlying spread are equivocal. Newer therapies such as tyrosine kinase inhibitors have shown promise in reducing metastatic relapse in trials, nonetheless, not all patients respond and 5-year overall survival remains at ~ 50%. Better understanding of potential bone sarcoma biological subgroups, the role of the tumour immune microenvironment, factors that promote metastasis and clinical biomarkers of prognosis and drug response are required to make progress. In this review, we provide a comprehensive overview of the approaches to manage paediatric patients with metastatic Ewing sarcoma and osteosarcoma. We describe the molecular basis of the tumour immune microenvironment, cell plasticity, circulating tumour cells and the development of the pre-metastatic niche, all required for successful distant colonisation. Finally, we discuss ongoing and upcoming patient clinical trials, biomarkers and gene regulatory networks amenable to the development of anti-metastasis medicines.

<h4>Background</h4>Endocannabinoids are small lipid molecules that have critical roles in cellular proliferation and function. They are produced locally with their concentrations controlled via the endocannabinoid system (ECS). The important cellular functions of endocannabinoids have made them and the proteins that modulate their expression targets of potential interest for treatment in many different diseases including gynaecological conditions. There is significant evidence of heredity differences in the response to both exogenous and endogenous cannabinoids that hampers the identification of effective targets. Whether compounds targeting endocannabinoids will be effective therefore may rely on personal differences mediated through genetic architecture. To investigate the source of individual differences, we investigated the effects of genetic variants on the expression of the endocannabinoid system genes at both a systemic and individual tissue level with a particular focus on the female reproductive system and the endometrium.<h4>Methods</h4>We performed this analysis using publicly available datasets, including the 31,684 participants from the eQTLGen database and 838 donors to the GTEx database which includes 49 different sources of tissue, as well as an in-house database of 206 endometrial samples. Analysis of the eQTLGen data identified 22,020 eQTLs that influenced 43 of the selected 70 ECS genes.<h4>Results</h4>A comparison across 49 different tissues that included at least 70 different individuals in the GTEx dataset identified eQTL for 69 of the 70 different genes, confirming a tissue-specific influence. Comparisons among 11 different physiological system indicated that the female reproductive system was associated with a fewer number of eQTLs. Finally, in the endometrium, we detected Bonferroni significant genetic effects on one individual gene fatty acid binding protein 3 (FABP3), an intracellular transporter that delivers endocannabinoids to the enzyme responsible for its inactivation, with a further 14 independent FDR significant eQTL for 13 ECS genes.<h4>Conclusions</h4>This is the first study to investigate the effects of genetic variants on the ECS gene transcription and indicates genetic variants have significant influence that are unique to each tissue. Our results highlight the effect of individual variation and the impact endocannabinoid based therapies may have on different tissue and physiological systems.

<h4>Background</h4>Thermoregulation is critical for preventing hyperthermia during physical activity. In Huntington's disease (HD), research on possible thermoregulatory alterations has received little attention.<h4>Objectives</h4>We aimed to investigate and compare skin temperature differences before and after exercise between HD patients and healthy controls. Additionally, we aimed to evaluate potential correlations between thermoregulation, body composition, and HD severity.<h4>Methods</h4>This was a cross-sectional study. HD severity was assessed using the Total Functional Capacity and Unified HD rating scales. Skin temperature was assessed using thermographic infrared imaging of standardized 13 anatomical locations on the trunk and upper and lower extremities at baseline and after a treadmill test. Resting oxygen consumption was measured by indirect calorimetry. Differences between groups and correlation analyses between alterations in skin temperature and body composition, and HD severity were performed.<h4>Results</h4>We included 14 patients with mild-moderate HD, mean age 60.5 (32-68) years, and 10 healthy-matched controls, mean age 56 (29-65) years. Although, similar thermoregulation capacity was observed between HD patients and controls, uniquely in HD, reduced temperature recovery after exercise was inversely correlated with body mass index (particularly fat mass), resting oxygen consumption, and CAG repeats, and positively correlated with age and quality of life (mental health domain).<h4>Conclusion</h4>Although thermoregulation appears to remain largely intact in patients with mild to moderate HD, it tends to decline with aging, CAG repeat expansion, and lower BMI, which may negatively impact overall well-being in HD patients. However, further research with larger sample sizes is essential to confirm these results.

Transposable elements (TEs) constitute a significant part of plant genomes and shape their genomic landscape. While some TEs are ubiquitously dispersed, other elements specifically occupy discrete genomic loci. The evolutionary forces behind the chromosomal localization of TEs are poorly understood. Therefore, we first review specific chromosomal niches where TEs are often localized including (i) centromeres, (ii) (sub)telomeres, (iii) genes, and (iv) sex chromosomes. In the second part of this review, we focus on the processes standing behind non-equal distribution of various TEs in genomes including (i) purifying selection, (ii) insertion site preference or targeting of TEs, (iii) post-insertion ectopic recombination between TEs, and (iv) spatiotemporal regulation of TE jumping. Using the combination of the above processes, we explain the distribution of TEs on sex chromosomes. We also describe the phenomena of mutual nesting of TEs, epigenetic mark silencing in TEs, and TE interactions in the 3D interphase nucleus concerning TE localization. We summarize the functional consequences of TE distribution and relate them to cell functioning and genome evolution.

A long-standing question in biology is the extent to which cells function autonomously as opposed to requiring interactions with other cells or environmental factors. Here, we develop a framework to use interspecies chimeras to precisely decompose evolutionary divergence in any cellular trait into cell-intrinsic and cell-extrinsic components. Applying this framework to thousands of gene expression levels in reciprocal rat-mouse chimeras, we found that most divergence is cell intrinsic, though extrinsic factors also play an integral role. For example, cell-extrinsic regulation of a transcription factor can propagate to its target genes, leading to cell-type-specific extrinsic regulation of both their mRNA and their protein levels. We also show that imprinted genes are dramatically misexpressed in chimeras, suggesting a mismatch between rapidly evolving intrinsic and extrinsic imprinting mechanisms. Overall, our conceptual framework opens up new avenues to investigate the mechanistic basis of the evolution, development, and regulation of myriad cellular traits in any multicellular organism.

Neuronal loss is the ultimate driver of neural system dysfunction in Alzheimer's disease (AD). We used single-nucleus RNA sequencing and neuropathological phenotyping to elucidate mechanisms of neurodegeneration in AD by identifying vulnerable neuronal populations and probing for their differentially expressed genes. Evidenced by transcriptomic analyses and quantitative tau immunoassays of human AD and non-AD brain tissue, we identified a neuronal population especially vulnerable to tau pathology. Multiplexed immunohistochemistry and in situ hybridization (CBLN2 and LINC00507) validated the presence of the tau-vulnerable neuronal population and revealed a propensity of this population to bear tau pathology. Differentially expressed genes associated with phospho-tau pathology in these neurons revealed genes involved in apoptosis, cell-component dissociation (e.g., autophagosome maturation and actin filament depolymerization), and regulation of vesicle-mediated transport.

<h4>Objectives</h4>Triphenylphosphine (TPP) and Doxorubicin (DOX) were conjugated to obtain Triphenylphosphine-Doxorubicin (TPP-DOX), which was applied in tumor cells for enhancement of DOX in mitochondria targeting. The study focused on investigating the anti-tumor effect of TPP-DOX in combination with radiotherapy throughout <i>in vitro</i> and <i>in vivo</i> studies.<h4>Methods</h4>TPP-DOX was synthesized using the carbodiimide method. <i>In vitro</i> experiments were conducted with 4T1 cells (mouse breast cancer cell line) to assess apoptosis induction, mitochondrial targeting, reactive oxygen species (ROS) production, and mitochondrial membrane potential. The research evaluates the effects of TPP-DOX, DOX, and their combinations with radiotherapy. A nude mouse tumor heterograft model was established to investigate the synergistic effect of TPP-DOX and radiotherapy.<h4>Results</h4>TPP-DOX was successfully synthesized and scrupulously verified. <i>In vitro</i> experiments showed that compared to DOX, TPP-DOX exhibited enhanced tumor cytotoxicity, improved cellular uptake in 4T1 cells, and increased apoptosis induction. Combined with radiotherapy, TPP-DOX promoted mitochondrial ROS production, reduced mitochondrial membrane potential, and amplified its anti-tumor effect. <i>In vivo</i> experiment confirmed that TPP-DOX combined with radiotherapy exhibited superior anti-tumor activity, promoted tumor tissue apoptosis, inhibited tumor angiogenesis, and showed a favorable <i>in vivo</i> safety profile.<h4>Conclusion</h4>The study confirmed that when combined with radiotherapy, TPP-DOX promoted tumor cell apoptosis, and effectively enhanced the anti-tumor effect. In sensitive cells, TPP-DOX demonstrates comparable efficacy to DOX when combined with radiotherapy. TPP-DOX holds significant potential for a broader spectrum of applications and emerges as a valuable candidate for clinical application. These findings provide a promising and efficient therapeutic strategy for tumor treatment with improved efficacy and safety.

Approximately 70% of people will experience a traumatic event in their lifetime, but post-traumatic stress disorder (PTSD) will only develop in 3.9% and complex post-traumatic stress disorder (CPTSD) in 1-8% of the population worldwide, although in some countries (e.g., Poland and Northern Ireland) it will develop in a much higher percentage. Stress-related disorders have a complex pathogenesis involving neurophysiological, genetic, epigenetic, neuroendocrine and environmental factors. This article reviews the current state of knowledge on the molecular aspects of selected PTSD symptoms: hypervigilance, re-experiencing, emotion dysregulation and dissociation, i.e., the symptoms with strong neurobiological components. Among analysed susceptibility factors are specific gene polymorphisms (e.g., <i>FKBP5</i>, <i>COMT</i>, <i>CHRNA5</i>, <i>CRHR1</i>, <i>5-HTTLPR</i>, <i>ADCY8</i> and <i>DRD2</i>) and their interactions with the environment, changes in the HPA axis, adrenergic hyperactivity and disturbances in the activity of selected anatomical structures (including the amygdala, prefrontal cortex, corpus callosum, anterior cingulate gyrus and hippocampus). It is worth noting that therapeutic methods with proven effectiveness in PTSD (TF-CBT and EMDR) have a substantial neurobiological rationale. Molecular aspects seem crucial when searching for effective screening/diagnostic methods and new potential therapeutic options.

Depression and breast cancer (BC) demonstrate significant clinical comorbidity, yet their shared molecular mechanisms remain unclear, particularly regarding immune pathway regulation. This study systematically analyzed Depression-associated gene expression profiles (Gene Expression Omnibus (GEO) database) and BC transcriptomic data (The Cancer Genome Atlas (TCGA) database), identifying overlapping differentially expressed genes (DEGs). Functional enrichment (Gene Ontology (GO)/Kyoto Encyclopedia of Genes and Genomes (KEGG)) and protein-protein interaction (PPI) network analyses (STRING/Cytoscape) were employed to elucidate biological processes, followed by least absolute shrinkage and selection operator (LASSO) regression and receiver operating characteristic (ROC) curve validation to prioritize key genes. Immune infiltration patterns were assessed via the xCell algorithm, with Spearman correlation linking genes to immune subsets, and single-gene Gene Set Enrichment Analysis (GSEA) evaluating pathway activity. In total, 93 overlapping genes were identified, with predominant involvement in immune-related pathways being revealed by functional enrichment analysis. <i>BHLHE41</i>, <i>EpCAM</i>, and <i>GSTM2</i> were prioritized as mechanism-associated genes through integrated LASSO regression and ROC analyses. Significant correlations were observed between these genes and specific immune cell populations. GSEA further linked these genes to immune response pathways, suggesting their regulatory roles. These findings highlight immune dysregulation as a shared mechanism underlying Depression-BC comorbidity, providing a foundation for developing early diagnostic strategies and therapeutic strategies targeting both conditions.

Moyamoya disease (MMD) is a rare yet clinically significant cerebrovascular disorder characterized by progressive stenosis of the distal internal carotid artery and/or its principal branches, accompanied by the development of characteristic collateral vessel networks. This disease demonstrates a complex multifactorial etiology with strong genetic determinants, as evidenced by its distinct geographical distribution patterns and familial clustering. Recent genetic researches have identified multiple pathogenic mutations contributing to MMD development through three principal mechanisms: progressive vascular stenosis, abnormal angiogenesis, and dysregulated inflammatory responses. Furthermore, moyamoya syndrome frequently occurs as a secondary vascular complication in various monogenic disorders. This review provides a comprehensive analysis of recent genetic advances in MMD in view of diverse pathogenic pathways, offering valuable perspectives on the molecular mechanisms underlying disease development and potential therapeutic targets.

Atrial fibrillation (AF) is the most prevalent sustained arrhythmia, associated with significant morbidity, mortality, and healthcare burdens. Despite therapeutic advances, recurrence rates remain high, particularly in persistent AF, underscoring the need for deeper mechanistic insight. Epigenetic regulation-comprising DNA methylation, histone modifications, chromatin remodeling, RNA methylation, and non-coding RNAs-has emerged as a key contributor to the structural, electrical, and inflammatory remodeling underlying AF. These mechanisms operate at the interface of genetic susceptibility and environmental exposure, offering a dynamic framework for understanding disease progression. Systemic stressors such as aging, obesity, diabetes, hypertension, hypoxia, and alcohol have been shown to induce epigenetic reprogramming in atrial tissue, further promoting atrial cardiomyopathy and arrhythmogenesis. Additionally, circulating epigenetic markers, particularly microRNAs, are being investigated for their potential in AF diagnosis, risk stratification, and therapeutic monitoring. Therapeutic strategies targeting epigenetic pathways-ranging from histone deacetylase inhibitors and miRNA-based therapeutics to CRISPR/dCas9-mediated epigenome editing-are under investigation. Additionally, sodium-glucose cotransporter 2 inhibitors may indirectly influence epigenetic programs and miRNA expression relevant to atrial remodeling. While promising, these approaches require further validation in terms of safety, delivery specificity, and long-term efficacy. High-resolution epigenomic mapping and integrative multi-omic approaches may enhance understanding of AF heterogeneity and enable personalized treatment strategies. This review provides an integrated appraisal of epigenetic mechanisms in AF and outlines their emerging diagnostic and therapeutic relevance.

Yunong Black (YN) pigs and Yunong Black × Landrace (YL) hybrid pigs exhibit significant differences in meat quality characteristics. Studies have suggested that extrachromosomal circular DNA (eccDNA) may play a regulatory role in muscle development. In order to study the differences in eccDNA between two groups with different meat quality traits and their potential biological significance, this study used the Circle-seq method to detect eccDNA in the longest dorsal muscle (LDM) of Yunong Black pigs (YN) (<i>n</i> = 3) and Yunong Black × Landrace hybrid pigs (YL) (<i>n</i> = 3). EccDNA-related differentially expressed genes (eccDEGs) were then analyzed in combination with RNA-seq to explore the mechanisms by which eccDNA affects meat quality. The results showed that 1325 and 1304 differentially expressed eccDNAs were identified in the YN and YL groups, varying in size and distributed across multiple genomic functional regions. These eccDNAs were also annotated according to several protein-coding genes. Combined analysis with RNA-seq results revealed 19 and 27 eccDEGs in the YN and YL groups. The Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) analysis enriched many lipid-related pathways, such as chemokine signals and ADP metabolic processes. By constructing a regulatory network, several potential regulatory networks that might be related to pork quality, for example, ecc_sus_8665/ssc-miR-212/ADAMTS16, were identified. In summary, we identified several potential eccDNAs that may regulate pig muscle, offering insights into the regulation of pig muscle traits for breeding.

Triple-negative breast cancer (TNBC) is a subtype of breast cancer associated with poor prognosis and limited targeted treatment options. Lipid metabolism plays a pivotal role in the initiation, progression, and metastasis of TNBC by supporting cancer cell energy production, facilitating membrane biosynthesis, and regulating signal transduction. Dysregulation of lipid metabolism promotes tumor cell proliferation and contributes to processes such as epithelial-mesenchymal transition (EMT), angiogenesis, and immune evasion. Targeting lipid metabolism-such as inhibiting fatty acid synthase (FASN) and lipid metabolic byproducts-has emerged as a promising therapeutic strategy. The integration of multi-omics approaches and advanced imaging technologies can further elucidate the interactions between lipid metabolism and the tumor microenvironment, thereby supporting precision oncology. Future research should explore the role of lipid metabolism in distinct TNBC subtypes, optimize therapeutic strategies, and improve patient outcomes, particularly for those who are unresponsive to conventional treatments.

Copper, an essential trace element for the human body, plays a key role in energy metabolism, mitochondrial respiration, redox reactions, and neural signal transmission. The recently proposed concept of "cuproptosis" has further revealed the unique status of copper in cellular regulation: when copper abnormally accumulates within cells, it can directly bind to the lipoylated proteins of the mitochondrial TCA cycle, triggering protein aggregation and metabolic disorders, ultimately leading to cell death. This form of cell death plays an important role in various neurodegenerative diseases of the central nervous system, such as Alzheimer's disease (AD), Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS), Huntington's disease (HD), and stroke. This review summarizes recent research on the mechanisms of cuproptosis, providing new perspectives and a theoretical basis for understanding the pathogenesis of these neurodegenerative diseases.

This article analyzes the progress of animal experiments on the analgesic mechanism of electroacupuncture (EA) at the central level for neuropathic pain (NP) in the past 10 years, and summarizes the analgesic mechanism of EA at the central level for NP. EA, as a safe and reliable treatment, can treat NP by regulating the release of nociceptive neurotransmitters and receptors, upregulating the expression of non-coding RNA (ncRNA), inhibiting the activation of microglia, Ca²⁺/calmodulin kinase II (CaMKII) phosphorylation, dendritic spine remodeling, endoplasmic reticulum stress (ERS), and glucose metabolism. NP is a type of pain caused by various diseases. Pain caused by stroke, spinal cord injury, postherpetic neuralgia (PHN), diabetes, and chemotherapy-induced neuropathy all fall into the category of NP, which makes the treatment of NP very challenging. At present, EA research on the treatment of NP is more focused on the mechanism of the dorsal horn of the spinal cord, and there are relatively few animal experiments at the level of the central brain region. There is also a lack of clinical trials using human subjects and relevant biochemical indicators. In the future, electrophysiology, neuron tracing, and multi-omics techniques combined with emerging technologies such as artificial intelligence should be used to further improve the analgesic mechanism of EA on the central level for NP, making EA the best treatment for NP.

<h4>Objective</h4>To identify cell senescence-related genes and immune cell infiltration characteristics in intervertebral disc degeneration (IVDD) using bioinformatics and investigate their biological functions and signaling pathways.<h4>Methods</h4>GSE56081 and GSE23130 datasets were downloaded from the Gene Expression Omnibus (GEO) database. The former was used for the analysis of differentially expressed genes (DEGs) and the latter was used as the validation set. Intersection analysis of DEGs and cell senescence-related genes was performed to screen for senescence-related differentially expressed genes (SRDEGs). SRDEGs were analyzed using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). A protein-protein interaction (PPI) network was also drawn, and the hub SRDEGs were obtained using 11 different algorithms. Immune infiltration analysis was then performed. Receiver operating characteristic (ROC) curve and quantitative polymerase chain reaction (qPCR) were used to evaluate the diagnostic value of the hub SRDEGs.<h4>Results</h4>Four hub SRDEGs, SP1, FOXO1, ESR1, and MAPK1, were identified. SP1 was downregulated in IVDD, while the other three hub genes were upregulated. ROC curve verification showed that the AUC for SP1, FOXO1, ESR1, and MAPK1 were 0.483, 0.683, 0.683, and 0.725, respectively. RT-qPCR confirmed that MAPK1 expression was higher in the degeneration group (t = 3.229, P <0.001). Immune infiltration analysis demonstrated elevated proportions of CD8 T cells and activated NK cells in IVDD samples, with MAPK1 showing positive correlations with CD8 T cells, activated NK cells, and neutrophils but negative correlations with naive CD4 T cells, B memory cells, and resting NK cells.<h4>Conclusion</h4>These findings highlight MAPK1 as a pivotal regulator of cellular senescence and immune cell infiltration in IVDD pathogenesis, offering novel therapeutic targets for intervention.

<h4>Introduction</h4>The differentiation of hematopoietic stem cells (HSCs) into diverse blood and immune cells is a complex, highly hierarchical process characterized by a series of tightly regulated steps. It involves a sequence of intermediate oligo-potent progenitors making successive binary decisions. This process gradually narrows down lineage possibilities until a final fate is reached. This step-wise process is tightly controlled by transcription factors (TFs) and their associated regulome, ultimately resulting in the differentiation of both lymphoid and myeloid compartments.<h4>Methods</h4>We unravel the lineage-specific gene regulatory circuitry controlling the development of B cells, T cells, innate lymphoid cells (ILCs), and dendritic cells (DCs). We employ weighted gene co-expression network analysis to characterize gene modules associated with the lymphoid or myeloid cell fate, enabling the identification of lineage-restricted TFs based on their expression patterns.<h4>Results</h4>By identifying TFs whose expression is subset-restricted or those with a broader expression in the hematopoietic compartment, we construct a regulatory logic that potentially controls the development of these key immune cells. Our results point to conserved regulatory elements between ILCs, natural killer cells, and DCs. This analysis unravels an intricate relationship between each cell type and how the expression of key TFs dictates lineage specificity. We particularly dissect the elements associated with conventional DCs and plasmacytoid DCs.<h4>Discussion</h4>In conclusion, our findings shed new light on regulatory mechanisms controlling blood cell development and offer a blueprint that can be leveraged to better understand the molecular mechanisms underpinning blood cell development.

The depletion of ovarian reserve is a major factor contributing to the decline in female fertility. It is characterized by a simultaneous reduction in the quantity and quality of oocytes and the follicular pools. The cyclic recruitment of primordial follicles and the preservation of oocyte quality involve complex and tightly regulated biological processes. Granulosa cells, which surround the oocytes, play a pivotal role in follicular development and the determination of follicular fate. Programmed cell death (PCD), a genetically regulated process of cell elimination, is a key factor in the regulation of ovarian reserve dynamics. Emerging evidence suggests that natural products derived from medicinal plants, dietary components, animals, and microorganisms may modulate PCD in granulosa cells through various molecular mechanisms and signaling pathways. These natural products have demonstrated preliminary effects in delaying ovarian aging and preserving ovarian reserve in preclinical models. This review discusses the roles and underlying mechanisms of various forms of PCD in diminished ovarian reserve, while summarizing the current findings on natural products that influence granulosa cells PCD to protect ovarian function. These insights may contribute to the future development of novel, targeted strategies aimed at preserving female reproductive potential.

<h4>Objective</h4>Considering the high prevalence of metabolic dysfunction-associated steatotic liver disease (MASLD) among patients with type 2 diabetes and its associated complications, this study aimed to investigate the relationship between dietary acid load (DAL) and the risk of MASLD in patients with diabetes.<h4>Methods</h4>This cross sectional study was conducted on 200 patients aged 18 to 70 with type 2 diabetes. Of whom, 133 participants were diagnosed with MASLD based on transit elastography (Fibroscan). For biochemical evaluation of liver enzymes, lipid profile, and fasting blood sugar, venous blood samples were collected after 10-12 h of fasting. Dietary acid load was determined using a 147-item food frequency questionnaire based on PRAL (potential renal acid load) and NEAP (net endogenous acid production).<h4>Results</h4>A total of 108 women and 92 men with an average age of 52.2 years and an average body mass index of 28.8 kg/m² participated in the study. After adjusting for confounders, the risk of MASLD in the third tertile of PRAL was 3.1 times higher than the first tertile (OR = 3.1, 95% CI = 1.2-7.7). After adjusting for all confounding factors, participants in the highest tertile of NEAP had nearly seven times the chance of developing MASLD compared to those in the lowest tertile, which was statistically significant (OR = 7.3, 95% CI = 2.6-20.3). Overall, the data analysis revealed a significant direct relationship between both PRAL (P trend = 0.016) and NEAP (P trend < 0.001) with the risk of MASLD.<h4>Conclusion</h4>Our analysis revealed that a higher dietary acid load is associated with an increased risk of MASLD and liver steatosis in patients with type 2 diabetes.

The onset of obesity is characterized by early physiological and molecular changes, including leptin resistance and hypothalamic dysfunction, preceding significant weight gain and metabolic complications. Extracellular vesicles (EVs) are key mediators of intercellular communication, reflecting early pathological shifts in metabolic disorders. This study investigates the role of hypothalamic EVs (hEVs) in early obesogenic insult and their potential implications for obesity-related comorbidities. Using a hamster model fed a high fat diet for 30 days, next-generation proteomics revealed altered hEV protein compositions linked to cellular metabolism, neuroinflammation, and metabolic dysfunction, mirroring early obesity-related dysfunction. These findings highlight the adaptive molecular profiles of hEVs during early obesogenic insult and their potential as biomarkers and molecular mediators in obesity progression and its comorbidities. In conclusion, this study provides new insights into the molecular mechanisms underlying the onset of obesity and highlights hEVs as promising targets for early detection and intervention.

Glyphosate (N-phosphonomethylglycine) is a widely used organophosphorus herbicide that inhibits the shikimate pathway, a crucial metabolic route responsible for the synthesis of aromatic amino acids in plants and certain microorganisms. Due to its broad-spectrum activity, glyphosate serves as the main active ingredient in various commercial herbicide formulations, including Roundup and Kalach 360 SL (KL). It poses a health hazard to animals and humans due to its persistence in soil, water erosion, and crops. The aim of our study was to continue the previous research to explore the impact of KL on bone using physico-chemical parameters and in silico studies after exposing female <i>wistar</i> rats for 60 days. The in silico study concerned the assessment of binding affinity and molecular interactions using computational modeling approach. The rats were allocated into three experimental groups: group 1 (<i>n</i> = 6) served as controls, while groups 2 and 3 received low and high doses (Dose 1: 126 mg/Kg and Dose 2: 315 mg/Kg) of KL dissolved in water, respectively. All rats were sacrificed after 60 days of exposure. XRD and FTIR spectrum analysis of bone tissues in female rats showed significant histoarchitectural changes associated with bone mineralization disruption. Our results have demonstrated that sub-chronic exposure of adult female rats to KL causes bone rarefaction, as confirmed by a previous histological study. This physico-chemical study has further confirmed the harmful impact of KL on the crystalline fraction of bone tissue, composed of hydroxyapatite crystals. In addition, the computational analyses showed that glyphosate binds to 3 Glu form of osteocalcin (3 Glu-OCN) (4MZZ) and decarboxylated osteocalcin (8I75) with good affinities and strong molecular interactions, which justified and supported the in vivo findings. In conclusion, KL may interfere with hydroxyapatite and osteocalcin and, therefore, impair bone remodeling and metabolism.

Resveratrol is a naturally occurring phytoalexin found in red grapes, cocoa berries, and red grape wine. This compound exhibits potent antioxidant, anti-inflammatory, and anticancer properties. However, its clinical application is significantly hindered by poor aqueous solubility and rapid degradation at physiological pH, resulting in extremely low systemic bioavailability. This review explores three key aspects: green extraction methods for the efficient and sustainable isolation of resveratrol; structure-activity relationship studies of resveratrol derivatives to identify compounds with improved bioavailability and therapeutic efficacy; and advanced drug delivery systems to enhance resveratrol solubility, stability, and achieve targeted tissue delivery. All of these solutions collectively aim to increase resveratrol bioavailability, enabling the development of effective pharmaceutical formulations and maximizing the clinical potential of this promising compound. The aim of this review is to summarize the key studies published in the last five years, highlighting innovative advancements in sustainable extraction, structural modifications, and delivery strategies for resveratrol.

The IgLON family of tumor suppressor genes (TSG) impact a variety of cellular processes involved in cancer and non-cancer biology. OPCML is a member of this family and its inactivation is an important control point in oncogenesis and tumor growth. Here, we analyze RNA-Seq expression ratios in ovarian cancers from The Cancer Genome Atlas (TCGA) (189 subjects at Stage III) to identify genes that exhibit a cooperative survival impact (via Kaplan-Meier survival curves) with OPCML expression. Using enrichment analyses, we reconstruct functional pathway impacts revealing interactions of OPCML, and then validate these in independent cohorts of ovarian cancer. These results emphasize the role of OPCML's regulation of receptor tyrosine kinase (RTK) signaling pathways (PI3K/AKT and MEK/ERK) while identifying three new potential RTK transcriptomic linkages to KIT, TEK, and ROS1 in ovarian cancer. We show that other known extracellular signaling receptor ligands are also transcriptionally linked to OPCML. Several key genes were validated in GEO datasets, including KIT and TEK. Considering the range of OPCML impacts evident in our analyses on both external membrane interactions and cytosolic signal transduction, we expand the understanding of OPCML's broad cellular influences, demonstrating a multi-functional, pleiotropic, tumor suppressor, in keeping with prior published studies of OPCML function.

<h4>Introduction</h4>Several studies identified genetic factors and key cellular signaling associated with developing immunoglobulin A nephropathy (IgAN). However, there is still a lack of understanding regarding the relationship between hub-transcription factors (TFs) encoding genes and changes in immunogenic activity. Our objective was to identify hub-TF encoding genes associated with immune cell infiltrations, immunogenic pathway activity, and potential drug candidates in IgAN through bioinformatics techniques.<h4>Methods</h4>We utilized GSE104948, GSE93798, GSE115857, GSE37460, and GSE35487 to identify key significant DEGs and validation in IgAN relative to the control. Next, we employed various bioinformatics approaches to investigate the key genes and their relationship with immunity in IgAN. Finally, we identitified enriched drugs and elcudate their molecular interactions with TFs via molecular docking approaches.<h4>Results</h4>We identified 1,123 differentially expressed genes (DEGs) between IgAN and control samples, comprising 342 upregulated genes and 780 downregulated genes. The upregulated genes are linked to immune-related biological processes and KEGG pathways, while the downregulated genes are associated with metabolic processes. Five significant clusters were identified, enriched in several KEGG pathways. We explored 26 hub-TF encoding genes, including GATA2, HDAC1, TSC22D3, SOX9, RARA, RORA, KLF5, KMT2A, FOSB, and FOSL1, which were consistently dysregulated in IgAN patients. Immunogenic analysis revealed increased levels of Th1 cells, pDCs, monocytes, M2 macrophages, fibroblasts, endothelial cells, and activated dendritic cells in IgAN. The activity of various immunological pathways was also elevated. The expression of hub-TFs like GATA2, HDAC1, TSC22D3, SOX9, RARA, RORA, KLF5, KMT2A, FOSB, and FOSL1 correlated with immune signatures and pathways in IgAN. Additionally, these hub-TFs were linked to diagnostic efficacy and drug interactions. Molecular docking identified key drug candidates for inhibiting HDAC1 and modulating RARA, suggesting their potential for IgAN treatment.<h4>Conclusions</h4>We identified key hub-TFs and their association with immune infiltration and immune pathways linked to IgAN initiation and progression. These findings provide important insights into the immunological mechanisms driving IgAN and propose potential treatment approaches. Molecular docking further revealed key drug candidates for inhibiting and modulating these targets, highlighting their therapeutic potential for IgAN.

<h4>Summary</h4> Cells must coordinate DNA segregation with cytokinesis to ensure that each daughter cell inherits a complete genome. Here, we explore how DNA segregation and division are mechanistically coupled in archaeal relatives of eukaryotes, which lack CDK/Cyclins. Using live cell imaging we first describe the series of sequential changes in DNA organisation that accompany cell division in Sulfolobus , which computational modelling shows likely aid genome segregation. Through a perturbation analysis we identify a regulatory checkpoint which ensures that the compaction of the genome into two spatially segregated nucleoids only occurs once cells have assembly a division ring - which also defines the axis of DNA segregation. Finally, we show that DNA compaction and segregation depends, in part, on a ParA homologue, SegA, and its partner SegB, whose absence leads to bridging DNA. Taken together, these data show how regulatory checkpoints like those operating in eukaryotes aid high-fidelity division in an archaeon.

We previously identified a signature of 16 serum proteins that highlighted a role of the e2 allele of APOE in lipid regulation via apolipoprotein B (APOB) and apolipoprotein E (APOE), and in inflammation. The serum proteins were profiled using the aptamer-based Somalogic technology. Here, we validate and expand the serum protein signature of APOE using a combination of mass-spectrometry, ELISA, Luminex, antibody-based Olink proteomics, and blood transcriptomics. We replicate the association between APOB and the e2 allele of APOE, we correct the pattern of association between APOE genotypes and serum level of APOE, and we detect new associations between APOE genotypes and the complex of apolipoproteins APOC1, APOC4, APOC2, APOC3, APOE, APOF and APOL1. In addition, we discover 13 new proteins that correlate with APOE genotypes. This extended signature includes granule proteins CAMP, CTSG, DEFA3, and MPO secreted from neutrophils and points to olfactomedin 4 (OLFM4) as a new target for the prevention of Alzheimer’s disease.

To address the challenges of recycling and high-value utilization of waste fluororubbers, an effective method is reported for producing novel photocurable vinyl-terminated liquid fluororubbers (VTLF) with elevated fluorine content (63.1%), superior temperature resistance (T_10% = 335 °C) from commercial waste fluororubbers. The approach employs a streamlined, multifaceted system (oxidative degradation/condensation reaction) integrating microwave, mechanical, and steady-state temperature fields. This system facilitates both efficient recycling and high-value transformation of waste fluororubbers. Initially, waste fluororubbers undergo controlled/oxidative degradation induced by alkali and hydrogen peroxide to yield carboxyl-terminated liquid fluororubbers (CTLF). Subsequently, condensation reaction system efficiently converts carboxyl groups into photoreactive vinyl groups. Ultimately, environmentally friendly and efficient photocuring of VTLF is achieved. The nonthermal effects of microwave fields reduce the total process time to just 1 h. The resulting photocured VTLF exhibits not only the comprehensive properties of conventional fluororubbers but also excellent chemical stability and unique light transmittance (94.21%). This study proposes a green, straightforward upcycling strategy within the circular economy framework to mitigate environmental issues associated with rubber's covalent crosslinking. Furthermore, it opens avenues for designing and synthesizing novel fluoropolymers for diverse applications.

Cell cycle structures vary significantly across cell types, which exhibit distinct phase compositions. Asynchronous DNA replication and dynamic cellular characteristics during the cell cycle result in considerable heterogeneity in DNA dosage, chromatin accessibility, methylation, and expression. Nonetheless, the consequences of cell cycle disruption in the interpretation of multi-omics data remain unclear. Here, we systematically assessed the influence of distinct cell phase structures on the interpretation of omics features in proliferating cells, and proposed solutions for each omics dataset. For copy number variation (CNV) calling, asynchronous replication timing (RT) interference induces false CNVs in cells with high S-phase ratio (SPR), which are significantly decreased following replication timing domain (RTD) correction. Similar noise is observed in the chromatin accessibility data. Moreover, for DNA methylation and transcriptomic analyses, cell cycle-sorted data outperformed direct comparison in elucidating the biological features of compared cells. Additionally, we established an integrated pipeline to identify differentially expressed genes (DEGs) after cell cycle phasing. Consequently, our study demonstrated extensive cell-cycle heterogeneity, warranting consideration in future studies involving cells with diverse cell-cycle structures. RTD correction or phase-specific comparison could reduce the influence of cell cycle composition on the analysis of the differences observed between stem and differentiated cells.

The efficient establishment of in vitro neural models that accurately mimic the structural and functional connectivity of neural networks is critical in neuroscience research. 3D bioprinting shows great potential for constructing sophisticated in vitro models with high freedom of design. However, mature neurons are delicate and susceptible to manipulation. Here, extrusion-based 3D bioprinting is employed to fabricate gelatin methacryloyl (GelMA)-based constructs containing embryonic day 18 (E18) rat cortical neurons, referred to as 3D neuMatrix. 3D neuMatrix displays favorable neuronal viability, with the progressive formation of a 3D brain-like neural network with local and long-range functional axon connections. Compared with 2D cultured neurons, 3D neuMatrix is more similar to the E18 cortex according to the bulk transcriptomic profile, with a recreation of cellular components in the cerebral cortex. The 3D neuMatrix is employed to establish a disease model of ischemic stroke, with a faithful recapitulation of the viability, function, and transcriptomic features of rats with middle cerebral artery occlusion/reperfusion (MCAO/R). These findings demonstrate the formation of multiscale neural circuits within 3D neuMatrix and its valuable potential in the study of neurodevelopment, disease modeling with drug screening, and in vitro intelligence.

The eight-segmented RNA genome of influenza A virus (IAV) is transcribed and spliced into 10 major viral mRNAs in the nucleus of infected cells. Both transcription and splicing are facilitated by the host RNA polymerase II (Pol II) machinery via interactions between the viral ribonucleoprotein (vRNP) complex and various host factors. In this study, we demonstrate that IAV vRNPs recruit species-specific heterogeneous nuclear ribonucleoprotein M (hnRNPM) to support their replication in human and avian cells through distinct mechanisms. In A549 cells, human hnRNPM specifically facilitates the efficient transcription of HA, NA, M, and NS segments of WSN virus in a gene coding sequence-dependent manner. In contrast, in DF-1 cells, chicken hnRNPM restricts excessive splicing of M segment mRNA to ensure proper M2 protein production. Notably, human hnRNPM, with 34 additional amino acids compared with its chicken counterpart, fails to inhibit the M2 expression in DF-1 cells, whereas both human and chicken hnRNPM regulate WSN virus replication similarly in A549 cells. These findings highlight the host-specific roles of M2 levels in IAV replication and reveal how IAV co-opts host factors through virus genome sequence-dependent and host species-specific mechanisms, underscoring its high flexibility and adaptability during cross-species transmission.IMPORTANCEThe transcription and splicing of IAV genome in the nucleus of infected cells are precisely regulated to produce optimal amounts of viral proteins, ensuring efficient virus replication. In this study, we discovered that human hnRNPM regulates the IAV segment-specific differential transcription in a coding sequence-dependent manner in human cells. In contrast, chicken hnRNPM specifically inhibits M2 mRNA splicing to maintain proper M2 protein levels in avian cells. These species-specific regulatory mechanisms highlight the distinct replication strategies employed by IAV in human versus avian cells and underscore the complexity of cross-species transmission.

The influence of genetic ancestry on genomics in T-cell Acute Lymphoblastic Leukemia (T-ALL) has not been fully explored. We examined the impact of genetic ancestry on multi-omic alterations, survival outcomes, and risk stratification. Among 1309 children and young adults with T-ALL treated on the Children's Oncology Group trial AALL0434, the prognostic value of five commonly altered T-ALL genes varied by ancestry-including NOTCH1, which was associated with superior overall survival for patients of European ancestry but non-prognostic among patients of African ancestry. Integrating genetic ancestry with published T-ALL risk classifiers, we identified that a X01 Penalized Cox Regression classifier stratified patients regardless of ancestry, whereas a European multi-gene classifier misclassified patients of certain ancestries. Overall, 80% of patients harbored a genomic alteration in at least one gene with differential prognostic impact in an ancestry-specific manner. These data demonstrate the importance of incorporating genetic ancestry into genomic risk classification.

Bone morphogenetic protein (BMP) antagonists have been increasingly linked to the development of colorectal cancer (CRC). BMP signalling operates in opposition to the WNT signalling pathway, which sustains stem-cell maintenance and self-renewal of the normal intestinal epithelium. Reduced BMP and elevated WNT signalling lead to expansion of the stem-cell compartment and the hyperproliferation of epithelial cells, a defining characteristic of CRC. Chordin-like-2 (CHRDL2) is a secreted BMP antagonist, with overexpression linked to poor prognosis and variants in the gene shown to be associated with an elevated CRC risk. However, the detailed mechanism by which CHRDL2 contributes to CRC is unknown. In this study, we explored the impact of CHRDL2 overexpression on CRC cells to investigate whether CHRDL2's inhibition of BMP signalling intensifies WNT signalling and enhances the cancer stem-cell phenotype and response to treatment. Our research approach combines 2D cancer cell lines engineered to inducibly overexpress CHRDL2 and 3D organoid models treated with extrinsic CHRDL2, complemented by RNA sequencing analysis. CHRDL2 was found to enhance the survival of organoids and CRC cells during chemotherapy and irradiation treatment due to activation of DNA damage response pathways. Organoids treated with secreted CHRDL2 exhibited elevated levels of stem-cell markers and reduced differentiation, as evidenced by diminished villi budding. RNA-seq analysis revealed that CHRDL2 increased the expression of stem-cell markers, WNT signalling and other well-established cancer-associated pathways through BMP inhibition. These findings collectively suggest that CHRDL2 overexpression could affect response to CRC therapy by enhancing DNA repair and the stem-cell potential of cancer cells, and its role as a biomarker should be further explored.

Heparanase (HPSE) is the sole mammalian endoglycosidase that degrades heparan sulfate (HS) proteoglycans, disrupting the extracellular matrix (ECM) and promoting cancer invasion and metastasis. Although HPSE overexpression is linked to tumor progression, no clinically approved HPSE inhibitors exist. We developed aminoglycoside-based HS mimetics with defined sulfation and hydrophobic modifications to target HPSE's lipophilic pockets, a novel approach distinct from traditional HS glycans. Computational modeling showed that these mimetics engage HPSE through hydrophobic and π-π stacking interactions, enhancing affinity. The most potent compounds inhibited HPSE-driven ECM degradation, tumor cell proliferation, and invasion. <i>In vivo</i>, the lead candidate significantly reduced metastatic burden in B16 melanoma and MPC-11 myeloma models, showing tumor growth inhibition (TGI = 83.1%) <i>versus</i> SST0001 (TGI = 58.6%) and matching bortezomib. Importantly, the compound was well-tolerated with no notable toxicity. These results support HPSE as a cancer target and highlight aminoglycoside-based HS mimetics as promising therapeutics for metastatic cancer.

The intestinal mucosa undergoes a dynamic process of continual proliferation, differentiation, and apoptosis. Delineating the mechanisms involved in intestinal epithelial cell (IEC) differentiation is crucial to our understanding of not only normal gut adaptation but also aberrant intestinal growth. Bone morphogenetic protein (BMP) signaling is a pivotal regulator of intestinal proliferation and differentiation. However, the molecular underpinnings of the BMP pathway in this context are not entirely known. Here, we show a key role for the BMP4/microRNA (miR)-181/glycolysis signaling pathway in the maintenance of intestinal epithelial cell proliferation and differentiation. Treatment with BMP4 increased the expression of enterocyte markers and decreased proliferation of IECs, and importantly, decreased the expression of miR-181a-5p in mouse and human intestinal organoids. miR-181a-5p is a member of the miR-181 family with the highest expression in IECs. Treatment with locked nucleic acid (LNA) miR-181a-5p inhibitor significantly increased enterocyte differentiation as noted by increased expression of enterocyte markers in human and mouse intestinal organoids. In addition, LNA miR-181a-5p inhibitor repressed intestinal stem cell self-renewal as noted by the decreased organoid forming efficiency and expression of Ki67, cyclin D1, OLFM4 in human and mouse intestinal organoids. Moreover, in vivo administration of LNA miR-181a-5p inhibitor enhanced increased intestinal enterocyte differentiation and repressed intestinal cell proliferation. In contrast, overexpression of miR-181a-5p mimic decreased basal and BMP4-induced expression of enterocyte markers. Moreover, BMP4 treatment or inhibition of miR-181a-5p repressed hexokinase (HK) 1 expression and inhibited glycolysis. Consistently, knockdown of HK1 or inhibition of glycolysis using 2-deoxyglucose (2-DG) promoted enterocyte maturation and inhibited proliferation of IECs. Together, we provide evidence showing that miR-181a-5p inhibits intestinal enterocyte differentiation and promotes IEC proliferation through HK1-dependent glycolysis. Importantly, our findings identify miR-181a-5p as downstream in mediating BMP4 induction of enterocyte differentiation and inhibition of proliferation in IECs.

The choroid plexus (ChP) regulates cerebrospinal fluid (CSF) composition, providing essential molecular cues for brain development; yet, embryonic ChP secretory mechanisms remain poorly defined. Here we identify apocrine secretion by embryonic ChP epithelial cells as a key regulator of the CSF proteome and neurodevelopment in male and female mice. We demonstrate that the activation of serotonergic 5-HT_2C receptors (by WAY-161503) triggers sustained Ca²⁺ signaling, driving high-volume apocrine secretion in mouse and human ChP. This secretion alters the CSF proteome, stimulating neural progenitors lining the brain's ventricles and shifting their developmental trajectory. Inducing ChP secretion in utero in mice disrupts neural progenitor dynamics, cerebral cortical architecture and offspring behavior. Additionally, illness or lysergic acid diethylamide exposure during pregnancy provokes coordinated ChP secretion in the mouse embryo. Our findings reveal a fundamental secretory pathway in the ChP that shapes brain development, highlighting how its disruption can have lasting consequences for brain health.

C-reactive protein (CRP) reflects inflammation status and is linked to poor sleep, metabolic and cardiovascular health. Methylation (MRS) and polygenic risk scores (PRS) reflect long-term systemic inflammation, and genetically-determined CRP, respectively. To refine understanding of inflammation-linked sleep and health outcomes, we construct PRS-CRPs using GWAS summary statistics and a previously-developed MRS-CRP in the Hispanic Community Health Study/Study of Latinos. Via survey-weighted linear regression, we estimate associations between blood-, PRS-, and MRS-CRP, with multiple sleep and health outcomes (n = 2217). MRS-CRP and PRS-CRPs are associated with increasing blood-CRP level by 43% and 23% per standard deviation. MRS-CRP is associated with obstructive sleep apnea (OSA) traits, long sleep duration, diabetes and hypertension, while PRS-CRPs were not. Blood-CRP level is associated with sleep duration and diabetes. Adjusting for MRS-CRP weakens OSA-diabetes/hypertension associations. Consequently, MRS-CRP is a stronger marker than blood-CRP and PRS-CRP to systemic inflammation associated with poor sleep and related comorbidities.

Phase-separated p62 bodies activate NRF2, a key transcription factor for antioxidant response, by sequestering KEAP1, which targets NRF2 for degradation. Although p62 bodies containing KEAP1 are degraded by autophagy, they accumulate in various liver disorders. Their precise disease role remains unclear. We show that excessive KEAP1 retention in p62 bodies and NRF2 activation are major causes of liver damage when autophagy is impaired. In mice with weakened or blocked p62-KEAP1 interactions, KEAP1 retention and NRF2 activation under autophagy-deficient conditions were suppressed. Transcriptome and proteome analyses reveal that p62 mutants unable to bind KEAP1 normalize the expression of NRF2 targets induced by defective autophagy. Autophagy deficiency causes organelle accumulation, especially of the ER, regardless of p62 mutation. Liver damage and hepatomegaly resulting from autophagy suppression markedly improved in mice carrying p62 mutants, particularly those with blocked KEAP1 binding. These findings highlight excessive KEAP1 retention in p62 bodies and defective organelle turnover as key drivers of liver pathology, underscoring the significance of phase separation in vivo.

<h4>Background</h4>Metals in particulate matter (PM), like iron (Fe), were associated with lung injury. Inter-alpha-trypsin inhibitor heavy chain H4 (ITIH4) was suggested to inhibit lung inflammation. However, the effect of metals in PM, particularly Fe, on lung inflammation involving ITIH4 remained unclear.<h4>Methods</h4>We investigated the effects of recombinant ITIH4 (rITIH4) against acute lung injury in C57BL/6JNarl and B6.Sftpc-CreER^T2;Ai14(RCL-tdT)-D mice exposed to Fe-containing PM. Mice were exposed to diesel exhaust particles (DEP) or soluble iron (FeCl₃) via intratracheal instillation, while rITIH4 treatment was administered intranasally after exposure. Lung function, Fe levels (both bulk and single-cell by inductively-coupled plasma mass spectrometry (ICP-MS) and single-cell ICP-MS, respectively), inflammatory cell infiltration, and Hippo pathway regulation in type II alveolar epithelial cells (AECII) were assessed.<h4>Results</h4>We observed correlation between lung function changes and Fe levels, both in bulk and single-cell Fe in peripheral blood mononuclear cells. Single-cell RNA sequencing of the control group identified AECII-related cells characterized by high Sftpc, Sftpa1, Mzb1, B3 gnt5, Cacna1e, and Agbl1 expression. rITIH4 treatment in DEP-exposed mice restored Hippo pathway Cdh1, Itih4, Pdpn, Wwtr1, and Yap1 in AECII. rITIH4 reversed DEP- and Fe-induced increases in neutrophil infiltration, neutrophil-to-lymphocyte ratio, and monocyte depletion in bronchoalveolar lavage fluid (BALF). rITIH4 reduced BALF CXCL1/KC levels by DEP and serum 8-isoprostane levels by Fe. rITIH4 also reduced DEP-induced lung damage, increased ⍺-catenin and p-YAP in Fe-exposed mice, and pTAZ/TAZ ratio in both DEP- and Fe-exposed mice. rITIH4 increased pYAP/YAP ratio in DEP-exposed mice while decreasing LC3BII/I ratio in Fe-exposed mice.<h4>Conclusion</h4>ITIH4 attenuated acute lung injury in mice exposed to PM, specifically Fe, by modulating the Hippo pathway in AECII.

Stem cells differ from other somatic cells in that they possess self-renewal and differentiation potential, which endows them with unique characteristics, and have great therapeutic potential. Studies have shown that the self-renewal and differentiation potential of stem cells is regulated by ribosomes during protein synthesis. In this review, we discuss the translation regulation mechanisms and ribosome biogenesis in stem cells. Protein translation levels and ribosome biogenesis change dynamically during the development and differentiation of stem cells, and hierarchical translational regulation promotes stem cell differentiation. We also demonstrate that mitochondrial protein translation plays an important role in the regulation of stem cell fate. Ribosomes not only mediate the self-renewal and differentiation of stem cells through protein synthesis. They are also a key target for stem cell therapy. Understanding the mechanism of ribosome regulation in stem cells will allow better control of stem cells for their application.

<h4>Background</h4>Human papillomavirus (HPV) adversely affects human reproduction. We aimed to evaluate the prevalence of HPV infection in men and its correlation with semen parameters and reproductive outcomes.<h4>Methods</h4>In this prospective observational cohort study, 384 semen samples were collected from 237 male partners of infertile couples. The presence of HPV DNA and genotyping were analyzed in semen by quantitative PCR. A total of 186 intrauterine inseminations (IUI) in 101 couples and 186 assisted reproduction techniques (ART) cycles in 155 couples were performed. Associations between HPV positivity and semen parameters and fertility outcomes were evaluated using a generalized linear mixed model.<h4>Results</h4>The prevalence of HPV was 22.7%. Twenty-three HPV types were detected and 69.5% of positive samples presented at least one high risk (HR)-HPV genotype. HPV-18 (14%), HPV-53 (10%), and HPV-56 (10%) were the most prevalent HR-HPV genotypes followed by HPV-16, HPV-31, and HPV-51 (8%). HPV-42 was the most prevalent low risk (LR)-HPV genotype (25%). More than one HPV type was detected in 41% of HPV + samples. After capacitation, 30% of HPV + samples remained positive. We found no relationship between HPV infection and sperm volume, sperm concentration, and progressive motility both before and after semen capacitation. We observed a not significant different clinical pregnancy per cycle in the HPV - (6.8%) and HPV + (5.0%) IUI. We did not find any significant difference in fertilization, cleavage, quality of developed embryos, blastocyst formation nor in embryo utilization of ART cycles. Slightly lower cumulative pregnancy (33% vs 39%) and live-birth (25% vs 30%) rates and higher miscarriage rate (53% and 29%) were observed in HPV + with respect to HPV - cycles. Fifty-five neonatal outcomes from HPV - (n = 45) and HPV + (n = 10) cycles were available. No stillbirths as well as no malformations were recorded.<h4>Conclusions</h4>This study confirmed previous findings that HPV DNA is present in semen of one quarter of infertile couples. No significant association of seminal HPV presence with semen parameters was found. We observed a trend of worst clinical outcomes in the HPV + group that is worth further investigation in a large population to draw definitive conclusions.

To preserve functionality, bone is an active tissue that can constantly reconstruct itself through modeling and remodeling. It plays critical roles in the body, including maintaining mineral homeostasis, serving as the adult human body's core site of hematopoiesis, and supporting the structures of the body's soft tissues. It possesses the natural regeneration capacity, but large and complex lesions often require surgical intervention. Multiple omics integrate proteomics, metabolomics, genomics, and transcriptomics to provide a comprehensive understanding of biological processes like bone tissue injury and healing in bone tissue regeneration and engineering. Recently, bone tissue engineering and regenerative medicines have offered promising tools for bone regeneration using a multi-omics approach. Thus, this article will highlight the role of multiple omics in understanding bone tissue injury and healing. It will discuss the role of bone tissue engineering in developing bone substitutes that can replace translational medicine. Lastly, new developments in bone tissue engineering and regenerative medicine, along with multi-omics approaches, offer promising tools for bone regeneration.

Astragalus polysaccharide (APS) is the crucial active ingredient of Astragalus membranaceus, which has antioxidant, immunomodulatory and anti-inflammatory properties. However, the therapeutic effects and biological mechanisms of APS on chemotherapeutic intestinal mucositis (CIM) have not been clarified yet. Here, the protective mechanism and functional components of APS against CIM was investigated in both Drosophila melanogaster (fruit fly) and mice models. Administration of APS could remarkably attenuate the overall physiological impairments caused by CPT-11 in flies, including increased the survival rate, improved motility, restored the size of ovary and reproduction. APS supplementation could significantly alleviate CPT-11-induced intestinal damage, which involved in restoration of intestinal length, reduction of crop size and excretion, improvement of intestinal homeostatic imbalance, and restoration of intestinal shortened villi. Furthermore, the integration of transcriptomics and microbiomics demonstrated that APS exerted its protective effect mainly by mitigating oxidative stress associated with FoxO signaling, over-activated innate immunity and dysbiosis of intestinal flora. Subsequently, three molecular weight components (APS-I, APS-II and APS-III) were extracted from APS. Among the studied substances, APS-III as the lowest molecular weight demonstrated the highest efficacy in reducing intestinal mucositis compared to both APS-I and APS-II. Collectively, these results support that APS is intended to be constructed as an effective medication for addressing intestinal diseases.

An intervertebral disc is a pliable, cushioning structure present between the vertebrae in the spine. It functions as a shock absorber, minimizing friction and facilitating movement between the spinal bones. Degenerative changes are viewed as responses to injuries, whether mechanical or metabolic, rather than as distinct diseases. Degenerative disc disease is a frequent etiology of low back pain. Magnetic resonance imaging (MRI) commonly demonstrates disc desiccation, degeneration, herniation or protrusion, osteophyte formation, and facet joint arthropathy. In this case report we have specifically addressed a component of degenerative disc disease which is seen as an uncommon MRI finding of focal intradiscal fat. Although the direct impact of focal intradiscal fat on patient outcomes is not fully understood, it typically indicates advanced disc degeneration. Recognizing this finding can assist clinicians in diagnosing the extent of disc degeneration and developing appropriate treatment plans for patients with low back pain or related symptoms.

<h4>Background</h4>Although anaphylaxis is classically mediated by IgE, mast cells (MCs), and FcεRI, mouse experiments and clinical observations demonstrate an alternative pathway mediated by IgG, IgGFc receptors, and several cell types.<h4>Objective</h4>We sought to determine whether increased serum ferritin distinguishes IgG-mediated anaphylaxis from IgE-mediated anaphylaxis and to identify the mechanism for its increase.<h4>Methods</h4>Passive and active systemic anaphylaxis was induced, respectively, by injecting mice with an IgE or IgG anti-trinitrophenyl mAb and challenged with trinitrophenyl-ovalbumin, or by injecting mice with goat anti-mouse IgD antiserum and challenged with goat IgG. Readouts were hypothermia and hypomobility. Inhibitors were used to investigate cell types and molecules involved. Serum ferritin was quantitated by ELISA.<h4>Results</h4>IgG-mediated, but not IgE-mediated, anaphylaxis induced a severalfold increase in serum ferritin level beginning less than 0.5 hours after antigen challenge and lasting 24 to 36 hours. Polymorphonuclear leukocytes, MCs, and histamine contributed to the ferritin response, but histamine and platelet-activating factor were insufficient to induce the response, which was oxidation-dependent but shock-independent. Small ferritin responses were generated by mouse and human blood cells cultured with immune complexes and/or histamine.<h4>Conclusions</h4>IgG-mediated, but not IgE-mediated, anaphylaxis in mice is associated with a large, rapid, relatively long-lasting increase in serum ferritin that depends on neutrophils, MCs, oxidation, and histamine. Initial data suggest that immune complexes also increase ferritin in humans. Quantitation of serum ferritin should provide a way to detect IgG-mediated anaphylaxis and could help identify the optimal way to detect and treat anaphylaxis.

Cinnamic acid derivatives represent a significant class of biologically active compounds exhibiting a broad spectrum of activities, such as antifungal, antidengue, antimetastatic, antimicrobial, antibacterial, and anticancer properties. Their preparation has attracted considerable attention due to their versatile applications across the pharmaceutical, food, and chemical sectors. This review elucidates the functional groups of cinnamic acid that are instrumental in the rational design of biologically active derivatives. A comprehensive representative of recent advancements in synthetic methodologies over the past five years is presented, particularly emphasizing the active scaffolds of bioactive cinnamic acid derivatives. The review provides a strategic overview of alternative synthetic routes and highlights the latest innovations, including more efficient, highly selective, and environmentally sustainable approaches. Given the widespread incorporation of the cinnamic acid framework in various therapeutic agents, this review delivers critical insights into a molecular design for hit-to-lead optimization, offering detailed synthetic strategies for diverse functional modifications. By critically examining these methodologies, the paper underscores their role in expanding the utility of cinnamic acid derivatives and addressing prevailing challenges.

<h4>Objectives</h4>The genetic basis of calcium pyrophosphate deposition (CPPD) disease is unknown. This limits the development of therapeutic strategies. We aimed to analyse a genome-wide association study (GWAS) on a large administrative database to identify new candidate causal genes for CPPD disease.<h4>Methods</h4>We used publicly available GWAS summary statistics for Phecode-defined chondrocalcinosis and crystal arthropathy from the Veterans Affairs Million Veteran Program in people of African (AFR) and European (EUR) ancestry. Included were 3004 (536 AFR and 2468 EUR) cases for chondrocalcinosis and 3766 (700 AFR and 3066 EUR) cases for crystal arthropathy (operationally interpreted as calcium crystal arthropathy). Our primary analysis was in chondrocalcinosis, with secondary analysis in crystal arthropathy. We tested for colocalisation of chondrocalcinosis genetic association signals with genetic control of gene expression.<h4>Results</h4>There were 2 genome-wide significant loci for chondrocalcinosis in both AFR and EUR cases, both on chromosome 6 (signals within the ENPP1 and RNF144B genes). Findings were supported by analysis of the crystal arthropathy cohort. Colocalisation analysis of chondrocalcinosis genetic association signals with genetic control of gene expression and alternative splicing further supported ENPP1 and RNF144B as candidate casual genes. At ENPP1, the allele that increases the risk for chondrocalcinosis was associated with increased ENPP1 expression.<h4>Conclusions</h4>ENPP1 encodes ectonucleotide pyrophosphatase/phosphodiesterase family member 1 (NPP1) that produces adenosine monophosphate (AMP) and inorganic pyrophosphate which, together with calcium ions, leads to the formation of calcium pyrophosphate crystals. Selective NPP1 inhibitors developed for infectious disease and cancer could be tested as treatment for CPPD disease.

Alveolar rhabdomyosarcoma (ARMS) is a highly aggressive pediatric soft-tissue sarcoma driven by <i>PAX3/7-FOXO1</i> fusion proteins. Despite intensive multimodal therapy, outcomes remain poor for patients with fusion-positive ARMS. This review integrates recent advances in the molecular pathogenesis of ARMS, highlighting key diagnostic and therapeutic targets. We discuss the central role of fusion proteins in transcriptional reprogramming, impaired myogenic differentiation, and super-enhancer activation. Emerging biomarkers (<i>YAP</i>, <i>TFAP2B</i>, P-cadherin) and oncogenic kinases (Aurora A, <i>CDK4</i>, PLK1) are evaluated alongside receptor tyrosine kinases (<i>FGFR</i>, <i>MET</i>) and transcription factors involved in metabolic rewiring (<i>FOXF1</i>, <i>ETS1</i>). Additionally, we examine immunotherapeutic strategies, epigenetic modifiers, and noncoding RNAs as potential therapeutic avenues. Together, these insights provide a comprehensive framework for developing biomarker-guided, multi-targeted therapies to improve outcomes in ARMS.

Congenital hypogonadotropic hypogonadism (CHH) is a rare and heterogeneous genetic disorder with variable penetrance caused by GnRH deficiency, leading to delayed puberty and infertility. In 50-60% of cases, CHH is associated with non-reproductive abnormalities, most commonly anosmia/hyposmia (Kallmann syndrome, KS). Over 60 genes have been implicated in CHH pathogenesis. We aimed to perform genetic screening in a cohort of 14 patients (10 males, 4 females; mean age 22 ± 7.72 years) with suspected or diagnosed HH/KS. Genetic analysis was conducted using next-generation sequencing (NGS) with a custom panel of 46 candidate genes. Variant interpretation followed ACMG standards and guidelines. Multiple tools were used to predict the structural effects of variants on tertiary protein structure, assessing their pathogenicity. Novel variants were functionally characterized by qRT-PCR on mRNA extracted from peripheral leukocytes. NGS identified nine rare variants and four novel variants in genes previously associated with normosmic isolated HH (nHH) and/or KS (<i>FGFR1</i>, <i>PROK2</i>, <i>TAC3R</i>, <i>DCC</i>, <i>WDR11</i>, <i>IL17RD</i>, <i>DUSP6</i>, <i>KAL1</i>, <i>FGF8</i>, <i>IL17RD</i> and <i>DCC</i>). The variant in <i>TAC3R</i> (p.Trp275Ter) was pathogenic; variants in <i>ANOS1</i> (c.541+1G>A), <i>IL17RD</i> (c.1303_1304dup, p.Lys436ThrfsTer58), and <i>TAC3R</i> (p.Lys361Ter) were likely pathogenic. Nine variants were classified as variants of uncertain significance (VUS). Our study identified a possible genetic cause in 71% of the CHH/KS cohort, emphasizing the importance of genetic screening and functional characterization of genetic variants in patients with a phenotypically and genetically heterogeneous disorder like CHH.

Acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL) are genetically heterogeneous malignancies of hematopoietic stem cells, characterized by complex mutations and a high risk of drug resistance and relapse. Patient-derived xenograft (PDX) models are dynamic entities transplanted with leukemia stem cells (LSCs), retaining patients' biological and genetic characteristics. By elucidating LSCs, clonal dynamics, and microenvironment interaction, PDXs facilitate the preclinical evaluation of therapy sensitivity, including immunotherapies, epigenetic therapies, and other agents targeting mutated proteins or apoptosis. The application of PDXs has provided translational evidence for various studies with reliable clinical relevance. Additionally, conventional PDXs remain a robust tool in identifying drug resistance compared with other models, and their potential is further unleashed when examined in large cohorts or combined with novel technologies, which not only enhances our understanding of acute leukemia biology but also enables the discovery and identification of novel biomarkers. In this review, we present the application of PDX models for acute leukemia resistance, including mechanism investigation, therapy evaluation, and associated challenges.

<h4>Background</h4>Esophageal cancer (ESCA) is a highly aggressive malignancy characterized by poor prognosis, primarily due to late diagnosis and limited treatment efficacy. Immunotherapies, such as vaccines, necessitate a more comprehensive understanding of the tumor immune microenvironment and tumor-specific antigens. The immune heterogeneity of ESCA, which is shaped by immune cell infiltration and antigen presentation, remains largely unexplored, particularly in terms of its interactions with autoimmune mechanisms. This study employs multi-omics analysis to profile the immunophenotype of ESCA, aiming to identify immune evasion mechanisms, tumor antigens, and autoimmune-related pathways. By elucidating these features, we seek to uncover potential targets for vaccine development and personalized immunotherapies, thereby improving therapeutic outcomes.<h4>Methods</h4>To screen for potential antigen genes, we examined the overexpressed and mutated genes specific to ESCA. Additionally, we employed Kaplan-Meier survival and Cox analysis to evaluate the prognostic relevance of these potential tumor antigens. To achieve data aggregation and construct a consistency matrix, we used consistency clustering. Subsequently, a graph learning-based dimensionality reduction method was implemented to clarify the immune subtypes. Furthermore, weighted gene coexpression network analysis (WGCNA) was used to cluster potential antigen genes and identify hub genes.<h4>Results</h4>Our analysis identified six overexpressed and mutated tumor antigens that were strongly associated with poor prognosis and antigen-presenting cell (APC) infiltration in ESCA. Analysis of The Cancer Genome Atlas (TCGA) data consistently identified three immune subtypes. These findings allowed for the construction of the immune landscape of TCGA samples based on the respective immune subtypes. The integration of immunogenomics analysis further allowed for the characterization of the immune microenvironment for each immune subtype. WGCNA successfully screened for three prognostic factors.<h4>Conclusions</h4>In our analysis, <i>BTN2A1</i>, <i>MICA</i>, and <i>HHLA2</i> displayed significant potential as antigens for the development of anti-ESCA messenger RNA (mRNA) vaccines. The identification of three stable and reproducible immune subtypes specific to ESCA may prove essential in predicting the outcome of mRNA vaccines.

Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by inflammation and destruction of the joints due to the involvement of biologic, environmental, and genetic factors. Due to its pathogenesis being multifactorial in origin, the underlying molecular mechanisms contributing to the development of RA remain unclear. Therefore, understanding the factors driving RA is crucial for developing targeted therapies and improving patient outcomes. With various genetic variants contributing to RA, this article explores the role of differential gene expression in patients with RA and in different ethnic populations and how the genes contribute to RA susceptibility. Key takeaways from this review demonstrate how HLA shared epitope alleles and non-HLA genes have a strong association with RA and play an important role in immune regulation, autoantibody production, cytokine production, and development of extra-articular manifestations observed in RA. Additionally, gene expression in RA can vary across different sexes and ethnic populations, emphasizing the importance of developing personalized therapeutic interventions. These findings provide insight into the role of differential gene expression in improving diagnostic and therapeutic strategies and highlights potential therapeutic targets for RA management. Future research is needed to determine the clinical relevance of differential gene expression in developing interventions for RA treatment.

In this study, a graphene oxide (GO)/zinc oxide (ZnO)/hydroxyapatite (nHAp) composite coating was constructed on a pure titanium surface by microarc oxidation (MAO) pretreatment combined with hydrothermal technology (HT), thereby making it possible to explore the performance optimization of this coating for Ti-based implants. Scanning electron microscopy (SEM), an energy dispersion spectrometer (EDS), Fourier transform infrared spectroscopy (FTIR), Ramam spectroscopy (Ramam), etc., confirmed that the GO/ZnO/nHAp composites were successfully loaded onto the pure Ti surfaces. Through nanoindentation, differential thermal analysis (DiamondTG/DTA), and dynamic polarization potential detection, the GO/ZnO/nHAp composite coating imparts excellent nanohardness (2.7 + 1.0 GPa), elastic modulus (53.5 + 1.0 GPa), thermal stability, and corrosion resistance to pure Ti implants; hemolysis rate analysis, CCK-8, alkaline phosphatase (ALP) detection, alizarin red staining, and other experiments further show that the coating improves the hemocompatibility, biocompatibility, and bone guidance of the Ti implant surface. Studies have shown that GO/ZnO/nHAp composite coatings can effectively optimize the mechanical properties, corrosion resistance, biocompatibility, and bone guidance of pure Ti implants, so that they can obtain an elastic modulus that matches human bone.

CRISPR represent a groundbreaking genome-editing technology that has revolutionized genetic modification. This innovative tool offers an unparalleled revolution in the future treatment of genetic disorders, neurological diseases, infectious diseases and cancer. Despite the rapid expansion of CRISPR applications, its clinical use in humans is still relatively limited, with only 69 active clinical trials and 6 completed studies reported so far. This review examined current clinical trials and their processes in addressing various diseases via the CRISPR/Cas system. While earlier literatures have focused mainly on delivery methods and materials for CRISPR/Cas9, our review emphasized innovative targeting conditions and approaches for novel and functional therapeutic designs. In addition, we reviewed recent research to increase the efficiency of CRISPR editing in the management of genetic disorders and cancer, while exploring their future challenges and potential. This review provided a unique perspective on the advancement of CRISPR technology. By addressing these aspects, we aim to contribute to ongoing efforts to improve CRISPR-based therapies and expand their clinical applications, ultimately striving to transform the future of medical treatment.

Avian influenza H9N2 viruses cause significant economic losses to the poultry industry and pose a public health risk due to their potential to reassort with other avian influenza viruses, generating strains with zoonotic and pandemic potential. Two major H9N2 lineages dominate globally: the G1 lineage (genotype G1-B), prevalent in the Middle East, Africa and the Indian subcontinent, and the BJ/94 lineage (predominantly genotype G57), dominant in China, Vietnam, South Korea, Indonesia, and the Far East. We investigated replication, transmission, and pathogenicity of representatives of these two lineages, linking genotype to phenotype. The G57 strain A/Ck/Vietnam/H7F-14-BN4-315/2014 (Vietnam/315) was more lethal to chicken embryos than the G1-B strain A/chicken/Pakistan/UDL-01/2008 (Pakistan/UDL-01). Vietnam/315 exhibited higher replication in both directly infected and contact chickens, with increased virus shedding from the oropharynx and cloaca. In contrast, Pakistan/UDL-01 virus was primarily shed from the oropharynx, highlighting differences in replication, tissue tropism and transmission. Gene analysis showed the M gene of Vietnam/315 enhanced replication in primary chicken kidney cells, whereas the PB2, HA, NA, and M genes promoted increased replication in Madin-Darby Canine Kidney cells. Both viruses showed preferential binding to avian-like receptors over human-like receptors. However, Vietnam/315, however, exhibited higher neuraminidase activity and a more acid-stable HA (pH fusion 5.2) than Pakistan/UDL-01 virus. These findings suggest G57 genotype viruses possess greater replication and transmission fitness than G1-B viruses in vivo , ex vivo and in vitro . Reassortment events involving G1-B strains acquiring G57 genes may enhance replication and virulence, potentially increasing the risk of animal and human infection. <h4>Importance</h4> H9N2 avian influenza viruses are widespread in poultry, resulting in significant economic losses and occasional human infections. Different genetic variants dominate in various regions, but their ability to cause disease and spread in poultry remains unclear. These viruses can exchange genes with other avian influenza strains, altering their infectivity and transmission. This study compared two major H9N2 genotypes: G1-B (common in the Indian subcontinent and the Middle East) and G57 (dominant in China and Vietnam). The G57 virus showed higher replication in laboratory tests and infected chickens, shedding more virus through the orofecal route. It also exhibited a stronger attachment to bird cells and had a more acid-stable HA, suggesting an increased potential for infection and spread. Our findings indicate that G57 genotype viruses are more infectious and adaptable in poultry than G1 viruses. Gene exchange with other avian influenza strains may generate more virulent viruses with increased transmission potential. This study supports the risk assessment of emerging strains and enhances disease mitigation strategies.

Parkinson’s disease (PD) is a complex neurodegenerative disorder characterised by selective neuronal loss. We integrate deep full-length single-nuclei sequencing of the human substantia nigra with novel genome-wide association studies (GWAS) identifying genetic and cellular drivers of PD. Genetic risk converges on AGTR1+ dopaminergic neurons and perineuronal oligodendrocytes (pODCs), both reduced in PD, as well as oligodendrocyte precursor cells, enriched among disease-disrupted intercellular interactions. AGTR1+ neurons represent a metabolically stressed state, characterised by renin-angiotensin system (RAS) and MAPK activation, oxidative stress, and mitochondrial dysfunction, rather than a distinct subtype. AGTR1+ neurons and pODCs link PD risk to metabolic traits; in pODCs, this association reflects insulin resistance with downregulated PI3K–AKT signalling. GWAS of comorbid PD and type 2 diabetes (T2D) identifies loci in AGTR1 and TCF7L2, while AGTR1+ neurons specifically upregulate RAS and T2D drug targets. Familial PD genes associated to comorbid PD/T2D associate with non-Lewy body PD, stratifying disease mechanisms.

While progressive striatal gene expression changes and epigenetic alterations are a prominent feature of Huntington's disease (HD), the mechanistic basis remains poorly understood. Using chromatin immunoprecipitation and sequencing (ChIP-seq), we show that the huntingtin protein (HTT) reproducibly occupies specific locations in the mouse genome. Striatal HTT ChIP-seq peaks were enriched in coding regions of spiny projection neuron identity genes that were found to have reduced expression in HD patients and mouse models, and had reduced occupancy in expanded polyglutamine HTT knock-in mice (HttQ111/Q111). By contrast, HTT occupancy was depleted near genes that are upregulated in HD. ChIP-seq of striatal histone modifications revealed genotype-specific colocalization of HTT with active chromatin marks and enhancer of zeste homolog 2 (EZH2), a key enzymatic component of the PRC2 complex. In the vicinity of genes that are differentially regulated in HD, greater HTT occupancy in HttQ111/Q111 vs wild-type mice was associated with increased EZH2 occupancy, increased H3K4me3 levels and decreased H3K27me3 levels. Our study suggests that HTT-chromatin interactions may play a role in organizing chromatin and promoting cell type-specific gene expression, with HTT occupancy predicting transcriptional dysregulation in HD.

<h4>Background</h4>While neuropsychiatric symptoms are common in Huntington's Disease (HD), there is a dearth of evidence about the effectiveness of psychotropic medication for treating behavioral and cognitive symptoms. This article systematically reviews and aggregates the evidence of the effects of antidepressants on individuals with early HD.<h4>Methods</h4>A systematic review and random-effects meta-analysis of RCTs comparing antidepressants to placebo in individuals with HD was performed, with a focus on outcomes of executive functioning, functional capacity, mood, motor function, and adverse events.<h4>Results</h4>A total of 4 studies with 123 of patients with early HD, of whom a total of 63 (51.2%) received an antidepressant, were identified in our search. In our pooled analysis, a modest but statistically significant improvement in mood resulted from antidepressant treatment in HD (OR -1.22; 95% CI -2.16,-0.27; <i>p</i> = 0.02). No significant differences in the other outcomes of interest were found following antidepressant exposure.<h4>Conclusions</h4>Our investigation suggests that antidepressant use may modestly improve mood, but not ameliorate functional, cognitive, or movement-related symptoms in mild HD. Further studies involving novel agents, larger samples, and longer follow-up times are needed to better characterize the effect of antidepressants on neuropsychiatric symptoms in HD.

<h4>Background</h4>Asymmetry is a key feature of numerous developmental disorders and in phenotypic screens is often used as a readout for environmental or genetic perturbations. A better understanding of the genetic basis of asymmetry and its relationship to disease susceptibility will help unravel the complex genetic and environmental factors and their interactions that increase risk in a range of developmental disorders. Large-scale imaging datasets offer opportunities to work with sample sizes necessary to detect and quantify differences in morphology beyond severe deformities but also pose challenges to manual phenotyping protocols.<h4>Results</h4>We introduce a tool for quantifying asymmetry in 3D images and apply it to explore the role of genes contributing to abnormal asymmetry by deep phenotyping 3D fetal microCT images from knockout strains acquired as part of the Knockout Mouse Phenotyping Program. Four knockout strains: Ccdc186, Acvr2a, Nhlh1, and Fam20c were identified with highly significant asymmetry in craniofacial regions, making them good candidates for further analysis.<h4>Conclusion</h4>In this work, we demonstrate an open-source, semi-automated tool to quantify the asymmetry of craniofacial structures that integrates expert anatomical knowledge. This tool can detect abnormally asymmetric phenotypes in fetal mice to explore the relationship between facial asymmetry, perturbed development, and developmental instability.

Adolescence is a key period of development when major cognitive and neurobiological changes occur. Results from our lab showed that 5 days of social isolation in adolescent rats led to molecular changes in the Wnt/β-catenin pathway and to higher cocaine responses during adulthood. We assessed whether 5 days of social isolation (SI) during adolescence would impact on β-catenin levels in the prefrontal cortex (PFC) and nucleus accumbens (NAcc) as well as on anxiety-like behaviors in a sex- and time-dependent manner. We also investigated the role of dopaminergic neurotransmission on that impact, by using repeated administration of a D2 antagonist. Male and female Wistar rats were socially isolated between postnatal day (PND)30 to 35 or kept in their home cages (non-isolated), while they were treated with sulpiride (100 mg/kg, ip) or vehicle. Anxiety-like behaviors and exploratory activity were estimated by the open field test at 24 h (PND36) or 9 days (PND44) after isolation. Then, they were euthanized at PND36 or PND45, and β-catenin levels were analyzed by Western blot in PFC and NAcc. Our findings show that a brief SI during adolescence leads to a long-term impact on both β-catenin levels (10 days, PND45) and anxiety-like behaviors (9 days, PND44) with a significant increase and decrease, respectively, in female rats. In contrast, male rats show a rapid decrease in β-catenin levels in the PFC with no changes in anxiety-like behaviors (24 h, PND36). These suggest that adolescent SI induces mostly long-term changes in female while short term changes in male rats. Moreover, these changes seem to be modulated by dopaminergic neurotransmission since a sulpiride treatment during isolation prevented them.

Hepatocellular carcinoma (HCC) remains one of the most prevalent and lethal malignancies worldwide, with survival rates still falling short of expectations. Emerging evidence highlights the pivotal roles of both m6A methylation and ferroptosis-related genes (FRGs) in HCC progression. However, the prognostic significance of m6A-modulated FRGs remains largely unexplored. In this study, we developed a novel prognostic signature based on m6A-regulated FRGs, identifying six key genes (VEGFA, FANCD2, ZFP69B, EIF2S1, SLC7A11, and SRXN1) through multivariate and LASSO Cox regression analyses. A high m6A-FRGs score was strongly associated with poor prognosis, and multivariate analysis confirmed it as an independent prognostic factor. Notably, the high-risk group exhibited increased expression of immune checkpoint genes and a higher frequency of gene mutations. Functional assays further demonstrated that silencing ZFP69B significantly suppressed liver cancer cell proliferation, migration, and invasion. Clinical validation in 144 HCC samples revealed that elevated ZFP69B expression correlated with worse patient outcomes. Moreover, qPCR analysis confirmed CLSPN and HNRNPR as downstream targets of ZFP69B. Collectively, our findings establish the m6A-FRGs signature as a powerful prognostic tool for HCC and identify ZFP69B as a promising therapeutic target, warranting further investigation.

Upon injury, epithelial-derived IL-18 is released and induces an inflammatory response in underlying IL18R1⁺ lamina propria cells. Notably, <i>Il18r1</i> is also predicted to be expressed and functional in intestinal epithelial cells (IECs), since epithelial IL18R1 deficiency contributes to worsened outcomes upon inflammatory challenge. However, the nature of <i>Il18r1⁺</i> IECs, and their subsequent role in epithelial-intrinsic IL-18 signaling is poorly characterized. Here, we show that, in the murine small intestine, the IL-18 receptor is expressed by rare IECs that we identified to be a subset of enterochromaffin cells (ECC). While these cells are the major producers of serotonin in the intestine, we found no evidence that IL-18 regulated serotonin metabolism or release. Rather, upon radiation-induced injury, <i>Il18r1⁺</i> cells appeared in the crypt base and took on a revival stem cell (revSC) program, marked by mixed expression of YAP/TAZ and enteroendocrine genes signatures. Functionally, irradiated <i>Il18^-/-</i> mice display reduced epithelial proliferation and altered differentiation in the small intestine, characterized by increased Paneth cells (PC) and elevated <i>Wnt3</i> levels, which was partially recapitulated in <i>Il18^-/-</i> ileal organoids. In sum, we identified an <i>Il18r1</i>⁺ population in the epithelium and revealed a role for IEC-intrinsic IL-18 signaling during injury.

Obesity contributes to pulmonary dysfunction through poorly understood biochemical mechanisms. Chronic inflammation and altered cellular metabolism have emerged as pathological changes across organ systems in obesity, but whether similar changes occur in lungs with obesity is unknown. We collected bronchoalveolar lavage fluid (BALF) from right upper lobe and lingula pulmonary subsegments of 14 adults (7 males/7 females) with body mass indexes (BMIs) ranging from 24.3 to 50.9 kg/m² without lung disease. Proteomes were measured using sequential window acquisition of all theoretical fragment ion spectra (SWATH) mass spectrometry. Proteomic composition and pathway enrichments were examined for the cohort and as a function of BMI. BALF proteomic compositions were consistent with earlier studies and had improved protein identification. We found minimal differences in BALF proteomes between lavage regions. Five proteins were strongly correlated with BMI (False Detection Rate/FDR-adjusted <i>P</i> values < 0.05) and 11 had weaker correlation (FDR-adjusted <i>P</i> values 0.05-0.1). These proteins included acute phase reactants and complement factors. Few proteomic differences between biological sexes were detected, but some of them coincided with BMI-related proteins. Pathway enrichments impacted by BMI included innate immunity, antifibrinolysis, oxidative stress, and lipid metabolism. The bronchoalveolar microenvironment is altered by obesity in humans without lung disease. Pathway alterations associated with BMI included coagulation and fibrinolysis, redox and oxidative stress, energy metabolism, and humoral immune function. Our data support the theory that conserved biochemical and cellular changes in obesity may be fundamental mechanisms of dysfunction in multiple tissues but the specific impact on pulmonary function or disease is not yet known.<b>NEW & NOTEWORTHY</b> Obesity is thought to cause deleterious changes in lung biochemistry, but data in humans are lacking. We measured the alveolar proteome in bronchoalveolar lavages from subjects with a wide range of body mass index and no lung disease. We found changes in proteins and pathways associated with increasing body mass index that are similar to pathological changes observed in other tissues and may constitute mechanisms of pulmonary dysfunction in obesity.

This study used whole-genome sequence data on 406 beef cattle (203 Hanwoo and 203 Angus) to detect signatures of selection using four different methods; integrated haplotype score (iHS), Rsb, XP-EHH, and runs of homozygosity (ROH). Based on Rsb and XP-EHH analysis, 36 and 21 genomic regions differed significantly between Angus and Hanwoo breeds. Within breeds, we identified 108 regions (76 in Hanwoo and 32 in Angus) with the ROH analysis and 331 regions with the iHS method (298 in Hanwoo and 33 in Angus). The candidate genes related to meat quality, such as HSPA9 and LPL, were found within Hanwoo, while genes associated with growth and meat quantity traits, including ACTC1 and TMEM68, were identified within Angus. This study can assist in understanding the selection history of these breeds and identifying the genomic regions associated with the traits selected for in the breeding programs for these cattle breeds.

The skeletal muscle is highly susceptible to injury in daily life. Severe skeletal muscle injuries often result in incomplete regeneration, leading to functional impairment. In clinical practice, understanding the extent of skeletal muscle injury in limb trauma patients is crucial for selecting treatment modalities and assessing prognosis. Currently, there is a lack of specific indicators for evaluating the severity of mechanical skeletal muscle injury. Therefore, the aim of this study is to develop biomarkers for the early evaluation of different degrees of skeletal muscle injury. A rat model of skeletal muscle mechanical compression injury was established with varying degrees of injury severity, one control group, and two compression groups (Mild Injury and Severe Injury Group). LC-MS/MS-4D-DIA quantitative proteomics technology was used to detect the plasma proteome profile of rats in different injury groups at 3 hours post-injury, followed by bioinformatics analysis for data decoding. Rats in the mild and severe injury groups exhibited completely different degrees of injury and prognosis. The proteomic results of the plasma revealed that the relative quantification of 37 proteins increased along with the increase in injury, while 2 proteins decreased. These differentially expressed proteins (DEPs) included not only muscle-specific structural proteins but also metabolic-related proteins that might play crucial roles in tissue injury control, repair, and regeneration. Overall, the study has identified several potential protein biomarkers that can distinguish different degrees of skeletal muscle injury at an early stage. These protein biomarkers may be further developed to help clinicians identify patients with varying degrees of skeletal muscle injury, paving the way for personalized treatments.

<h4>Introduction</h4>Lipedema is a chronic and progressive disease that predominantly affects women, characterized by a disproportionate increase in subcutaneous adipose tissue (AT), particularly in the lower limbs. It is associated with significant physical disability, chronic pain, thromboembolism, and psychosocial distress. Despite its profound impact on women's health and quality of life, lipedema remains underrecognized and insufficiently studied, with an estimated prevalence of approximately 10% among women worldwide. Although the exact etiology of lipedema remains unclear, emerging evidence suggests a multifactorial origin involving genetic predisposition, hormonal influences, and vascular dysfunction-all contributing to its development and progression. Current therapeutic options provide only partial symptom relief and remain noncurative, highlighting the urgent need for expanded research and improved management strategies.<h4>Methods</h4>A systematic review was conducted to assess the current understanding of lipedema pathophysiology and current treatment options. Research articles were sourced from PubMed, Web of Science, ScienceDirect, and Scopus databases. Over 100 studies were incorporated.<h4>Results</h4>This review provides a comprehensive overview of lipedema, encompassing its clinical features, pathophysiological mechanisms, diagnostic challenges, and current treatment modalities. Additionally, the review discusses whether the molecular and metabolic differences between abdominal and femoral AT depots mirror those observed in classical obesity.<h4>Conclusions</h4>Multidisciplinary, research-informed care is essential for managing lipedema, combining conservative therapies, tailored exercise, and liposuction for advanced cases. More research to better understand the underlying pathophysiology is critical to developing targeted treatments, improving diagnostic accuracy, and informing standardized, evidence-based care.

Osteoarthritis is the most prevalent age-related degenerative joint disease and is closely linked to obesity. However, the underlying mechanisms remain unclear. Here we show that altered lipid metabolism in chondrocytes, particularly enhanced fatty acid oxidation (FAO), contributes to osteoarthritis progression. Excessive FAO causes acetyl-CoA accumulation, thereby altering protein-acetylation profiles, where the core FAO enzyme HADHA is hyperacetylated and activated, reciprocally boosting FAO activity and exacerbating OA progression. Mechanistically, elevated FAO reduces AMPK activity, impairs SOX9 phosphorylation, and ultimately promotes its ubiquitination-mediated degradation. Additionally, acetyl-CoA orchestrates epigenetic modulation, affecting multiple cellular processes critical for osteoarthritis pathogenesis, including the transcriptional activation of MMP13 and ADAMTS7. Cartilage-targeted delivery of trimetazidine, an FAO inhibitor and AMPK activator, demonstrates superior efficacy in a mouse model of metabolism-associated post-traumatic osteoarthritis. These findings suggest that targeting chondrocyte-lipid metabolism may offer new therapeutic strategies for osteoarthritis.

Severe type I antithrombin (AT) deficiency is considered to cause embryonic lethality. Although several pathological analyses using mice or zebrafish have been attempted, the previous studies did not unveil the detailed mechanism leading to lethality in the early developmental stage. In order to solve this problem, we established type I AT deficient zebrafish by the CRISPR/Cas9 system into Tg(gata1:dsRed) and Tg(fli1a:GFP) lines, so that we could conduct real-time imaging of thrombosis and angiogenesis using fluorescence stereo zoom microscopy. The established zebrafish AT (zAT) mutants harbored frameshift mutations which resulted to be type I AT deficient, unable to secrete zAT protein into blood. Both heterozygous (zAT^+/-) and homozygous (zAT^-/-) mutants showed reduced survival rate and diverse thrombosis up to 9 days post fertilization. In addition, blood vessel formation was delayed at 30 hpf in zAT^-/-, which was recovered normally by 5 dpf and had little effect on survival. Notably, we analyzed the differences in gene expression profiles under AT-depleted conditions by real-time quantitative PCR, and zAT^-/- juvenile zebrafish showed increased PLG gene expression and decreased F2 gene expression. Our in vivo study revealed the effects of AT deficiency on embryos during development from the aspects of coagulation and vascular formation.

S-methyl thioesters have proven to be useful intermediates in many biosynthetic reactions and their occurrence in bioactive molecules further enhance their appeal. Shortcomings in several reported synthetic protocols have led us to disclose herewith a mild, room temperature ex situ protocol that provides access to a large number of thioesters (aryl, heteroaryl, alkyl, amino acids). Late-stage functionalization strategies were also performed on commercial pharmaceutical drugs (containing carboxylic acid functionality) while further in situ conversion to valuable compounds has also been achieved.

The study of rare pediatric disorders is fundamentally limited by small patient numbers, making it challenging to draw meaningful biological conclusions. To address this, we developed a framework integrating clinical ontologies with proteomic profiling, enabling the systematic analysis of rare conditions in aggregate. We applied this approach to urine and plasma samples from 1140 children and adolescents, encompassing 394 distinct disease conditions and healthy controls. Using advanced mass spectrometry workflows, we quantified over 5000 proteins in urine, 900 in undepleted (neat) plasma, and 1900 in perchloric acid-depleted plasma. Embedding SNOMED CT clinical terminology in a network structure allowed us to group rare conditions based on their clinical relationships, enabling statistical analysis even for diseases with as few as two patients. This approach revealed molecular signatures across developmental stages and disease clusters while accounting for age- and sex-specific variation. Our framework provides a generalizable solution for studying heterogeneous patient populations where traditional case-control studies are impractical, bridging the gap between clinical classification and molecular profiling of rare diseases.

<h4>Background</h4>Successful restorative procedures, particularly for patients undergoing head and neck radiotherapy, rely on a robust adhesive interface. Since radiotherapy may alter dental tissues and compromise bonding, this study evaluated how the timing of radiation exposure affects the microtensile bond strength (µTBS) of a universal adhesive to dentin with different application modes.<h4>Methods</h4>Forty-two human third molars were used in this study. Mid-coronal dentin was prepared using a low-speed saw and polished with silicon carbide paper. The samples were randomly divided into 3 groups (n = 6) according to the timing of radiation exposure (no radiation, control; radiation after restoration, F-RT; and radiation before restoration, RT-F). A single radiation dose of 70 Gy was administered to the samples after restoration (F-RT) and before restoration (RT-F). The samples were further divided into 2 subgroups according to the application mode of a universal adhesive (Single Bond Universal adhesive, 3 M Oral Care, St. Paul, MN, USA): etch-and-rinse (ER) or self-etch (SE) mode. After adhesive application, the resin composite was constructed and stored in distilled water at 37 °C for 24 h. Six resin-bonded teeth per group were processed for the µTBS test. The data were analyzed by two-way ANOVA followed by Duncan's test (p < 0.05). Fractured surfaces were observed under a scanning electron microscope (SEM). Additional teeth were prepared for resin‒dentin interface observation (n = 1) and observed using an SEM.<h4>Results</h4>The application mode of the universal adhesive had no influence on the µTBS of dentin, regardless of irradiation (p = 0.670). The µTBS values of the control groups were not significantly different from those of the F-RT group but were significantly higher than those of the RT-F group for both application modes. The resin‒dentin interfaces of the samples were similar among the control, F-RT and RT-F groups. More abundant and longer resin tags were observed when the universal adhesive was bonded in ER mode.<h4>Conclusion</h4>Compared with radiation after restoration, radiation before restoration resulted in an inferior bond strength. The application mode of a universal adhesive had no effect on the bond strength.<h4>Clinical significance</h4>Restoration prior to the radiation procedure is an advisable strategy.

<h4>Background</h4>Eosinophilic endocarditis or myocarditis is a major complication of hypereosinophilic syndrome, characterized by eosinophilic infiltration leading to endocardial or myocardial necrosis, thrombosis formation, and fibrosis. For its rare morbility and various heterogenicity, eosinophilic endocarditis or myocarditis is prone to misdiagnosis and missed diagnosis. Neither large case series nor clinical trials on this specific endocarditis or myocarditis have been reported.<h4>Case presentation</h4>Four middle-aged male patients had increased eosinophilia and elevated levels of troponin or lactate dehydrogenase. Cardiac ultrasound showed ventricular wall thickening with or without reduced cardiac systolic function, apical thrombosis or restrictive cardiomyopathy.one of these patients showed myocardial enhancement by CMR, and one of these patients showed endocardial enhancement by CMR. The coronary angiography results were negative. Three patients were diagnosed with eosinophilic endocarditis, and one was diagnosed with eosinophilic myocarditis. After the application of steroid treatment, eosinophil levels decreased rapidly, myocardial thickening was relieved, and cardiac function was gradually recovered.<h4>Conclusion</h4>This case series embodies the high heterogeneity in the clinical manifestation of the eosinophilic myocarditis or endocarditis, and the important role of multi-module imaging. Early detection and early treatment is crucial for the prognosis of eosinophilic endocarditis or myocarditis.

<h4>Background</h4>Despite efforts to standardise practice using evidence-based guidelines, fresh frozen plasma (FFP) remains the blood component most frequently prescribed inappropriately. This study assessed the appropriateness of FFP transfusion in two tertiary teaching hospitals and analysed the characteristics of appropriate and inappropriate transfusions.<h4>Methods</h4>Patients who had undergone FFP transfusion between October and December 2022 at two tertiary teaching hospitals were retrospectively analysed. Only the initial FFP transfusion data were analysed for each patient. Patient characteristics; laboratory test results, including prothrombin time, international normalised ratio, and activated partial thromboplastin time; and the association of FFP transfusion with various factors were examined. Sub-therapeutic dosing was defined as the transfusion of ≤2 units of FFP. FFP transfusions were classified into eight groups based on a classification algorithm to determine their appropriateness.<h4>Results</h4>In total, 584 FFP transfusions (2301 units) were analysed, with 42.1% involving subtherapeutic dosing. FFP transfusions were performed in the intensive care unit (ICU; 30.5%), general ward (24.8%), operating room (21.1%), and emergency room (22.9%). Overall, 51.5% of FFP transfusions were deemed appropriate, with significant variations being observed between the hospitals (Hospital B vs. Hospital A: 73.2% vs. 35.3%). Inappropriate FFP transfusions were associated with a higher INR, with 73.4% of them being associated with severe bleeding and/or surgery.<h4>Conclusions</h4>In conclusion, 40.6% of FFP transfusions were deemed inappropriate in the present study owing to failure to meet laboratory criteria. The present study provides valuable insights into the optimisation of plasma transfusion practices and emphasises the requirement for institution-specific management.

Porphyria cutanea tarda (PCT) is caused by inherited or acquired defects of uroporphyrinogen decarboxylase (UROD) in the heme biosynthetic pathway. Altered iron homeostasis via hemochromatosis gene (HFE) mutations is one of many susceptibility factors associated with the sporadic form of PCT. Though sarcoidosis is not commonly associated with PCT, prior reports of hepatic sarcoidosis postulated that hepatic granulomas affect UROD activity by direct interference or immunological mechanisms. Here we describe a case of acquired PCT in the setting of (HFE):c.845G>A (p.Cys282Tyr) and (HFE):c.187C>G (p.His63Asp) compound heterozygosity, hepatic steatosis, and cutaneous sarcoidosis.

<h4>Introduction</h4>The incidence of recurrent implantation failure (RIF) can reach up to 10% among patients undergoing in vitro fertilisation and embryo transfer worldwide. However, the clinical efficacy of low-molecular-weight heparin (LMWH) in RIF remains a subject of controversy. Currently, there is a lack of high-quality clinical research validating the effectiveness of LMWH in treating patients with RIF, particularly during frozen embryo transfer (FET) cycles. Therefore, this randomised controlled trial aimed to investigate the impact of LMWH on pregnancy outcomes in women with RIF undergoing FET.<h4>Methods and analysis</h4>This prospective, single-centre, double-blind randomised, placebo-controlled clinical trial will be conducted in the Second Hospital of Shandong University, China. A total of 414 women with RIF, aged ≤40 years, who are undergoing FET cycles will be recruited and randomly assigned to the study group (LMWH) or the control group (placebo). Only one blastocyst which is from day 5 or day 6 and has a Gardner morphology score ≥4 BC will be transferred. LMWH 4000-6000 IU per day or placebo will be administered by subcutaneous injection from the day of transplantation. The primary outcome is the live birth rate. The secondary outcomes include the clinical pregnancy rate, biochemical pregnancy rate, embryo implantation rate, early miscarriage rate, ongoing pregnancy rate, ectopic pregnancy rate, pregnancy-related complications, perinatal complications, fetal birth weight, congenital malformations and other adverse reactions.<h4>Ethics and dissemination</h4>The protocol received approval from the Ethics Committee of the Second Hospital, Cheeloo College of Medicine, Shandong University (KYLL-2023-442). The findings will be disseminated in peer-reviewed publications.<h4>Trial registration number</h4>Chinese Clinical Trial Registry, ChiCTR2400083577.

Cells undergoing epithelial-to-mesenchymal transition (EMT) exhibit significant plasticity, making them more tumorigenic, invasive, and stem-like. PLCG2 has been identified as being linked to EMT. Specifically, the PLCG2-high subpopulation of tumor cells shows strong correlations with metastasis. However, it remains unclear whether PLCG2 serves as a direct driver of EMT. In this study, we employ an <i>in vivo</i> photostimulation method using tightly focused femtosecond-laser scanning to activate intracellular Ca²⁺ signaling and induce PLCG2 upregulation. By constructing a subcutaneous tumor model with prostate cancer PC3 cells and single-cell RNA sequencing, we identify distinct cell populations, including cancer stem cells, epithelial tumor cells, proliferating cells, and EMT cells. Upon photostimulation, EMT cells are notably expanded among the primary tumor cells, while epithelial tumor cells decrease in number. During the tumor progression, treatment with a specific PLCG2 inhibitor effectively suppresses the growth of the primary tumor but has no significant impact on metastatic cells. These findings offer valuable insights into the role of PLCG2 in regulating EMT and tumor development.

This study aimed to identify dysregulated proteins in Schirmer strip samples (ScS) from Sjögren's syndrome dry eye (SSDE) patients to uncover key biological processes using untargeted proteomics. The focus then shifted to two dysregulated proteins, PRDX6 and CXCL17, due to their roles in oxidative stress and mucosal immunity, with the goal of exploring their potential as therapeutic targets. Their involvement was further investigated in vitro and validated in a larger SSDE cohort, assessing their relationship with clinical signs. ScS from 12 SSDE patients and 6 healthy controls underwent untargeted proteomic analysis. PRDX6 and CXCL17 levels in ScS from 39 SSDE patients were quantified using ELISA. In vitro, human corneal epithelial cells (HCEc) were exposed to hyperosmolarity or IFN-γ, and PRDX6 and CXCL17 expression was assessed by RT-qPCR for gene expression and by ELISA and immunocytochemistry for protein expression. Untargeted proteomics identified 111 dysregulated proteins in SSDE, highlighting alterations in oxidative stress, cell metabolism, cytoskeleton organization, and programmed cell death. Targeted proteomics showed positive correlations between PRDX6 levels and TBUT/Schirmer tests, and negative correlations with OSDI/Oxford scores. CXCL17 levels negatively correlated with the Oxford score. In vitro, PRDX6 and CXCL17 expression increased under hyperosmotic or inflammatory stress, displaying inverse trends compared to ScS from SSDE patients. This study elucidates the biological processes driving epithelial cell alterations in SSDE, focusing on oxidative stress and mucosal homeostasis. It underscores the significant roles of PRDX6 and CXCL17 in these processes, suggesting their potential as biomarkers or therapeutic targets for SSDE.

Neurodevelopment is an intricate process with highly regulated, overlapping stages including neuronal differentiation and axon guidance. Aberrations during these and other stages are tied to the etiology of neurodevelopmental disorders like Autism Spectrum Disorder, Angelman Syndrome, and X-linked Intellectual Disability. Ubiquitination is a dynamic and highly reversible post-translational modification conferred by E3 ubiquitin ligases. Recent discoveries have advanced the understanding of how substrate ubiquitination can guide protein localization, drive protein degradation, and alter protein post translational modifications. In this review, we highlight members of the RING and HECT E3 ligase families to discuss their novel roles in the molecular mechanisms regulating neurodevelopment. These findings are both instrumental for informing the future directions of neurodevelopmental research, and in expanding knowledge of intracellular mechanisms of protein trafficking. In addition, a deeper understanding of the molecular mechanisms of E3 ligase function in development promises to offer new insights into the pathogenesis of neurodevelopmental disorders.

<h4>Background</h4>The most frequent malignant tumor in women is breast cancer (BRCA). It has been discovered that T-cell exhaustion and macrophages play significant roles in BRCA. It was necessary to explore prognostic genes associated with T-cell exhaustion and macrophage polarization in BRCA.<h4>Methods</h4>The following data were included: 35 macrophage polarization-related genes (MPRGs), 683 T-cell exhaustion-related genes (TEXRGs), GSE20685, as well as TCGA-BRCA. Initially, candidate genes were identified through crossing differentially expressed genes (DEGs) obtained by differential expression analysis, key module genes associated with MPRGs, as well as TEXRGs. Next, 101 combinations of 10 machine learning algorithms and univariate Cox analysis were utilized to screen for prognostic genes. Concurrently, a risk model was built for validation in TCGA-BRCA and GSE20685. Next, we conducted immune infiltration, immunotherapy, mutation analysis, molecular regulatory network, as well as drug sensitivity between the two risk groups. Ultimately, we did the reverse transcription-quantitative polymerase chain reaction (RT-qPCR).<h4>Results</h4>According to random survival forest (RSF) algorithm (the best combination with the greatest C-index of 0.799), 7 prognostic genes were selected, which are PGK1, BTG2, TANK, CFB, EIF4E3, TNFRSF18, and BATF. After that, we created a risk model, and in the low-risk samples, there was a relatively high survival rate. Next, between two risk parts, the 7 differential immune cells were found. There was a significant difference in 25 immunological checkpoint (ICI) genes between the two risk parts. Next, a lncRNAs-miRNA-mRNA network with 65 nodes and 70 edges was built. Additionally, 84 medications were shown to differ significantly between the two risk groups. Finally, the expression of BTG2, TANK, and EIF4E3 was verified by RT-PCR, which was consistent with the bioinformatics analysis.<h4>Conclusion</h4>The 7 prognostic genes (PGK1, BTG2, TANK, CFB, EIF4E3, TNFRSF18, and BATF) were screened, providing new insights into potential treatments for BRCA.

<h4>Introduction</h4>Lung adenocarcinoma (LUAD) is among the most prevalent and lethal forms of cancer worldwide, largely due to the lack of early symptoms and frequent late-stage diagnosis. G protein γ subunit 7 (GNG7) has been implicated in the regulation of cell proliferation, apoptosis, and migration across various cancers. However, its immunological role in LUAD progression remains poorly understood.<h4>Methods</h4>We analyzed The Cancer Genome Atlas (TCGA) database to assess the relationship between GNG7 expression and clinical outcomes in LUAD. Immune cell infiltration and immune-related gene expression were evaluated in association with GNG7 levels. In vitro functional assays, including proliferation, migration, invasion, and apoptosis assays, were performed following GNG7 overexpression. A prognostic model was constructed based on immune-related genes regulated by GNG7 and validated using the GSE31210 and IMvigor210 cohorts.<h4>Results</h4>Low GNG7 expression was associated with enhanced tumor growth and poor prognosis in LUAD patients. GNG7 expression correlated significantly with immune cell infiltration and key immune regulatory markers. In vitro, GNG7 overexpression suppressed LUAD cell proliferation, migration, and invasion, while promoting apoptosis. The developed GNG7-related immune gene prognostic model effectively predicted both patient prognosis and immunotherapy response.<h4>Discussion</h4>Our findings highlight the critical role of GNG7 in LUAD progression and its modulation of the tumor immune microenvironment. GNG7 shows promise as a prognostic biomarker and potential therapeutic target for immune-based LUAD treatment strategies.

It is unusual for 2 distinct thyroid tumors to coexist, and even rarer to observe more than 2 thyroid pathologies alongside a malignant lymph node process. This report describes a highly rare presentation of multiple thyroid diseases coexisting with an extra-thyroidal malignancy. A 32-year-old female presented with suspicious thyroid nodules and bilateral pathological lymphadenopathy on ultrasound. Fine-needle aspiration (FNA) suggested both papillary thyroid carcinoma (PTC) and Hodgkin lymphoma (HL). Histopathology confirmed multifocal papillary thyroid microcarcinomas (PTMC), hyalinizing trabecular tumor (HTT), Hashimoto's thyroiditis (HT), and HL in a cervical lymph node. Managing such cases is clinically challenging due to their rarity and variable presentations. Further research is needed to guide evaluation and treatment strategies.

Meat quality traits, particularly WHC and tenderness, are pivotal for consumer satisfaction and economic value in the sheep industry. However, their genetic regulatory mechanisms remain unclear. We used RNA-Seq and WGCNA to identify genes regulating WHC and tenderness. Sixty longissimus thoracis samples were classified into high/low WHC (HWHC vs. LWHC) and high/low tenderness (HTN vs. LTN) groups. Comparative transcriptomics identified 270 differentially expressed genes (DEGs) linked to WHC, enriched in pathways like the regulation of the ATP metabolic process and the inhibition of canonical Wnt signaling. Key DEGs (e.g., <i>SORBS1</i>, <i>FOXO1</i>, <i>PDE4B</i>, <i>CDH1</i>) correlated significantly with WHC-associated traits. For tenderness, 165 DEGs were identified, including <i>LEP</i>, <i>FABP4</i>, <i>PLIN1</i>, and <i>GLP1R</i>, enriched in PPAR signaling, fat cell differentiation, and cAMP signaling pathways. WGCNA revealed modules associated with WHC and tenderness, with hub genes (<i>ATP2C1</i>, <i>GSKIP</i>, <i>PATL1</i>, <i>PPARA</i>, <i>CYLD</i>) involved in ATP metabolism, lipid biosynthesis, and myofibril assembly. Tissue-specific gene integration prioritized muscle-enriched candidates (<i>METTL21C</i> and <i>ACTC1</i>) with strong trait correlations. Our findings unveil interconnected gene networks governing WHC and tenderness, highlighting some candidate genes as potential biomarkers for precision breeding. This study provides novel insights into the molecular determinants of meat quality, offering actionable targets to enhance mutton production sustainability and consumer appeal.

Novel derivatives of valproic acid with biologically active moieties, such as thiomorpholine, 4-aminopyridine, serine methyl ester, trolox and the cinnamic acid derivative [(<i>E</i>)-3-(3,5-di-<i>tert</i>-butyl-4-hydroxyphenyl)acrylic acid], were synthesized at satisfactory yields. The conjugation of these moieties was based on the rationale of design and evaluation of compounds with selected structural characteristics, aiming at derivatives with multiple targets. These compounds reduced acute inflammation considerably and, in most cases, more than several highly used, well-known, non-steroidal anti-inflammatory drugs. They also offered the inhibition of soybean lipoxygenase, and some of them (compounds <b>5</b> and <b>6</b>) possessed radical scavenging and lipid peroxidation attenuating effects. Their antioxidant capacity was several times higher than that of the established antioxidant trolox. All the tested compounds decreased plasma lipid markers in tyloxapol-induced hyperlipidemia in rats. Compound <b>2</b> resulted in 71.1%, 52.8% and 79.1% decrease in total cholesterol, triglycerides and LDL-cholesterol, respectively, at 150 μmol/kg (<i>i.p</i>.). The effect on total and LDL cholesterol is comparable or equal to that of simvastatin, a hypocholesterolemic 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA) inhibitor, however, with additionally great triglyceride-decreasing effect compared to simvastatin. Thus, the synthesized compounds may be a valuable addition to multi-functional agents acting against various degenerative disorders that implicate inflammation and lipid derangement.

In this study, twenty-four new furoxan and seco-coumarin hybrids were synthesized, and their antiproliferative activities against four breast cancer cells (MCF-7/ADR, MCF-7, MDA-MB-231, and MDA-MB-468) were evaluated. Among them, compound <b>9e</b> exhibited significant toxicity against MCF-7/ADR cells compared to MCF-7 cells, with a 1401-fold increase, indicating its high collateral sensitivity. Meanwhile, <b>9e</b> exhibited relatively lower toxicity to normal cell lines and improved solubility compared to the previous active compound, <b>4A93</b>, which features a coumarin integrity core. Preliminary pharmacological studies revealed that <b>9e</b> might be a potential P-glycoprotein substrate, which enters the lysosomes of MCF-7/ADR to release effective concentrations of nitric oxide, producing reactive oxygen species and inducing apoptosis. Moreover, laser confocal microscopy and Western Blot experiments showed that <b>9e</b> could induce autophagy in MCF-7/ADR cells. Additionally, the anti-tumor activity of compound <b>9e</b> could be inhibited by the ferroptosis inhibitor Fer-1. These results suggest that the remarkable antiproliferative potency of these hybrids in MCF-7/ADR may be related to multiple anticancer mechanisms. As a novel nitric oxide donor, compound <b>9e</b> was used to explore the potential development of an anti-tumor candidate with special pharmacological mechanisms to overcome multidrug resistance in breast cancer.

Osteoarthritis (OA) is a degenerative disease with a series of metabolic changes accompanied by chondrocyte apoptosis. Chondrocytes express multiple receptors for neurotrophin, however, the role of neurotrophin receptor in chondrocyte metabolism remains unelucidated. Here, we first clarify the role of neurotrophin 3 (NT3) and its receptor tropomyosin receptor kinase C (TrkC) of chondrocytes in OA pathogenesis, using inducible TrkC-deficient mice (TrkC^fl/fl; Col2a1-CreER^T2 mice). Our findings show that TrkC levels are decreased in the chondrocytes and cartilage of patients with OA and OA-model mice. Chondrocyte-specific TrkC deficiency aggravates cartilage destruction during OA development. However, intra-articular TrkC-overexpressing adeno-associated virus (AAV) injection delays experimental OA progression. TrkC deficiency leads to decreased anabolic and increased catabolic activities in chondrocytes and stimulates chondrocyte apoptosis, thereby accelerating OA progression. Whereas TrkC overexpression rescues the imbalance between extracellular matrix synthesis and degradation and chondrocyte apoptosis through PI3K/Akt signaling. NT3, a multifunctional protein with high affinity for TrkC, effectively protects against cartilage degeneration in OA models <i>in vitro</i> and <i>in vivo</i> and relieves pain sensitivity in mice with OA. Our results indicate that TrkC is crucial for maintaining cartilage homeostasis and OA progression. Targeting TrkC with NT3 could be a novel strategy for OA treatment.

Structural variants (SVs) contribute to the genetic architecture of autism spectrum disorder (ASD), but their comprehensive characterization is limited by technological challenges in their detection. Optical genome mapping (OGM) offers a promising alternative, enabling the identification of large-scale SVs that might be overlooked by traditional sequencing methods. This study aimed to use OGM to identify SVs associated with ASD. We generated in-house OGM data from 26 participants diagnosed with ASD, leading to the discovery of 1593 novel SVs. Among them, 114 novel SVs were identified in at least two non-sibling participants, with 57 of them putatively overlapping known gene regions. To validate our findings, two novel SVs were confirmed by Sanger sequencing. The dataset generated in this study can serve as a novel and valuable resource for future research and facilitate the exploration of SVs related to ASD. Our work also underscores the importance of large-scale genomic rearrangements in neurodevelopmental disorders and provides insights into SVs as potential molecular diagnostic and therapeutic targets for ASD.

Featured milk generally refers to less common dairy products with unique sources or specific nutritional components and functional characteristics. We used omics methods to study the lipids, metabolites, and proteins in donkey milk, mare milk, and camel milk and compared them with human milk and pig milk. The phospholipid content in camel milk is relatively high. Donkey milk contains metabolites that have potential therapeutic effects on mental disorders and chronic inflammatory diseases. Camel milk exhibits potential neuroprotective effects. Compared with other species, human milk contains more receptor protein-tyrosine kinase and galectin-3-binding protein; camel milk contains more monocyte differentiation antigen CD14 and fibrinogen beta chain; while pig milk contains more aminopeptidase. This research provides a theoretical basis for the application of featured milk in the field of functional foods and other food sectors.

Owing to its exceptional physicochemical and biological properties, graphene oxide (GO), the oxidized form of graphene, has attracted considerable interest in bone regenerative engineering. The oxygen-functional groups on the backbone of GO enable biomolecule adherence, protein adsorption, cell adhesion, proliferation, differentiation, calcium ion adsorption and bone matrix mineralization. These oxygen functional groups enhance GO's interaction with biological fluids, facilitating its hydrolytic biodegradation. Recent preclinical studies have indicated that GO effectively improves mechanical strength, immunomodulation, and osteoinduction when utilized within diverse matrix structures including natural and synthetic polymers and ceramics to induce osteogenesis. Advanced bone regenerative applications of GO, such as implant coating and delivery of bioactive compounds, have demonstrated enhanced osseointegration, antibacterial efficacy, and pro-healing microenvironments. However, there are still challenges regarding the high-quality large-scale synthesis and long-term biocompatibility of GO. Additionally, the variability in the characteristics of GO resulting from different synthesis methods demonstrates further challenges for therapeutic translation. This study provides a comprehensive review of the recent preclinical research on the translational potential of GO, discussing the convergence of its exceptional properties for use in bone regenerative engineering along with its current challenges and future perspectives.

Strontium-added bioactive glass (SBG) has been widely used in bone tissue engineering. SBG can be prepared by conventional high-temperature melt-quenching or calcining sol-gelled powder at 700 °C or above. In the present study, the effects of calcination temperature (400-650 °C) and the amount of strontium addition (0-7 mol.%) were investigated simultaneously. The sol-gel process and post-calcination were used to prepare the Sr-added 58S bioactive glass (SBG) powders. The bioactivity of the SBG powder was assessed by immersing it in simulated body fluid, while biocompatibility and cytotoxicity were evaluated using L929 and MG63 cells, and a zebrafish animal model. The calcination temperatures were determined by thermogravimetric analysis based on the weight loss at various stages. X-ray diffraction was used to reveal the crystalline structure of calcined or SBF-immersed SBG powders. Meanwhile, the texture characteristics of SBG powders were examined by the BET method. Fourier-transformed infrared spectroscopy and scanning electron microscopy were used to investigate the absorption bands and powder morphology of SBG powders before and after SBF immersion. The experimental results showed that all SBG powders were mesoporous with a high specific surface area larger than 200 m²/g. SBG powder calcined at 650 °C with 5% Sr addition possessed a major Ca_14.92(PO₄)_2.35(SiO₄)_5.65 phase, the smallest pore size of 5.86 nm, and the largest specific surface area of 233 m²/g. It was noncytotoxic and exhibited good bioactivity and biocompatibility.

Methyldopa is a centrally acting antihypertensive agent commonly used in the management of hypertension. It has been associated with rare but serious hepatotoxic effects. Methyldopa-induced hepatitis is an uncommon but potentially devastating adverse effect that can mimic autoimmune hepatitis (AIH) both clinically and histologically. This report presents the case of a 31-year-old woman with a history of hypertension who developed acute hepatitis following the initiation of methyldopa. The patient presented with fatigue, jaundice, and elevated liver enzymes approximately six weeks after starting therapy. Viral, autoimmune, and metabolic causes of hepatitis were excluded through comprehensive testing. Liver function normalized following the discontinuation of methyldopa, and the patient recovered fully, supporting a diagnosis of drug-induced liver injury (DILI). Causality was assessed using the Roussel Uclaf Causality Assessment Method (RUCAM), yielding a score of 9, indicating a 'highly probable' link between methyldopa and the observed hepatitis.

We previously identified sex-specific genetic loci associated with memory performance, a strong Alzheimer’s disease (AD) endophenotype. Here, we expand on this work by conducting sex-specific, cross-ancestral, genome-wide meta-analyses of three cognitive domains (memory, executive functioning, and language) in 33,918 older adults (57% female; 41% cognitively impaired; mean age=73 years) from 10 aging and AD cohorts. All three domains were comparably heritable across sexes. Genome-wide meta-analyses identified three novel loci: a female-specific language decline-associated locus, VRK2 (rs13387871), which is a published candidate for neuropsychiatric traits involving language ability; a male-specific memory decline-associated locus among cognitive impaired, DCHS2 (rs12501200), which is a published candidate gene for AD age-at-onset; and a sex-interaction with baseline executive functioning, AGA (rs1380012), among cognitive impaired. We additionally provide evidence for shared genetic architecture between lifetime estrogen exposure and AD-related cognitive decline. Overall, we identified sex-specific variants, genes, and pathways relating to three cognitive domains among older adults.

The abundance and composition of transposable elements (TEs) varies widely across species, yet the evolutionary forces shaping this diversity remain poorly understood. Using 285 recently published genomes from drosophilid species, we investigated the evolutionary origins of the ≈130 TE families present in D. melanogaster and found that 79 were exchanged via horizontal transposon transfer (HTT) with other drosophilids. Most HTT events involved closely related species such as D. simulans, D. mauritiana , and D. teissieri , although transfers from more distantly related taxa were also observed. Notably, D. melanogaster appears to be a net recipient of HTTs, acquiring about three times as many TEs as it donated. Geographic patterns reveal that most HTTs involved Afrotropical species, reflecting D. melanogaster ‘s ancestral range, with fewer involving species from the Neotropics, a region which D. melanogaster invaded only ≈200 years ago. Despite colonizing the Nearctic, Australasian, and Palearctic regions between 200–2000 years ago, we found no evidence of HTT with species from those areas. Nonetheless, an analysis of drosophilids from each biogeographic realm shows that HTT is widespread in each realm, with 3–55% of the genome in each species derived from HTT. Strikingly, a considerable portion of the genome is shared among all species inhabiting the same realm —regardless of phylogenetic distance—indicating that geographic overlap, rather than shared ancestry, is a primary driver of TE composition. These findings highlight biogeography as a major force shaping the TE landscape and underscore the importance of ecological interactions in genome evolution.

Parkinson’s disease (PD) is associated with extensive structural brain changes. Recent work has proposed that the spatial pattern of disease pathology is shaped by both network spread and local vulnerability. However, only few studies assessed these biological frameworks in large patient samples across disease stages. Analyzing the largest imaging cohort in PD to date (N = 3,096 patients), we investigated the roles of network architecture and local brain features by relating regional abnormality maps to normative profiles of connectivity, intrinsic networks, cytoarchitectonics, neurotransmitter receptor densities, and gene expression. We found widespread cortical and subcortical atrophy in PD to be associated with advancing disease stage, longer time since diagnosis, and poorer global cognition. Structural brain connectivity best explained cortical atrophy patterns in PD and across disease stages. These patterns were robust among individual patients. The precuneus, lateral temporal cortex, and amygdala were identified as likely network-based epicentres, with high convergence across disease stages. Individual epicentres varied significantly among patients, yet they consistently localized to the default mode and limbic networks. Furthermore, we showed that regional overexpression of genes implicated in synaptic structure and signalling conferred increased susceptibility to brain atrophy in PD. In summary, this study demonstrates in a well-powered sample that structural brain abnormalities in PD across disease stages and within individual patients are influenced by both network spread and local vulnerability.

<h4>Purpose</h4>The tumor microenvironment (TME) may provide a useful framework for understanding the heterogeneous behavior of growth hormone (GH) secreting pituitary adenomas. Although the interest in TME in somatotropinomas has increased exponentially over the last few decades, there is limited elucidation of its mechanisms, particularly in relation to genes expression involved in its regulation.<h4>Methods</h4>A retrospective, observational, single-center study was conducted on 85 subjects: 46 patients diagnosed with acromegaly and 39 controls. After DNA extraction, clinical exome sequencing was performed and genomic alterations were detected, classified, and filtered using a dedicated bioinformatics pipeline.<h4>Results</h4>5759 unique genetic variants were found in patients with acromegaly. 33 patients (72%) showed the presence of at least one pathogenic variant in at least one of the following genes: FANCD2, SPTA1, TYRO3, and ZNF335. The enrichment pathway analysis of mutated genes was performed and showed that these genes were included in the same genetic pathway called "regulation of lymphocyte activation" (GO:0051249). Inflammatory infiltrate was analyzed in histological samples in 26 patients. A significantly higher number of CD68 + macrophages (P-value = 0.008), a lower number of CD8 + T lymphocytes (P-value = 0.037) and a higher CD68 + macrophages/ CD8 + T-lymphocytes ratio (P-value = 0.004) were observed in patients with pathogenic variants of genes of "regulation of lymphocyte activation" pathway.<h4>Conclusion</h4>This study provides new insights into the genetic basis of the TME in somatotropinomas and suggests that genetics may influence immune cells infiltration in acromegaly.

Chemically modified short interfering RNAs (siRNAs) unequivocally represent a groundbreaking class of drugs. The deliberate chemical modification of the natural structure has been pivotal to their resounding success. Specific modifications at certain positions bolster their potency, safety, stability, and specificity. In clinical research, 2'-<i>O</i>-methyl and 2'-fluoro are the most used modifications. The effects of a wide range of chemical changes in fully modified siRNAs have not been thoroughly evaluated for tolerability. In this study, we utilized two sequences in a fully modified siRNA to systematically assess the tolerability of single nucleotide backbone and sugar modifications, including deoxyribonucleic acid, 2'-<i>O</i>-(2-methoxyethyl), locked nucleic acid, unlocked nucleic acid, mismatches, butane diol substitution, and butane diol insertion. We synthesized 522 siRNA variants and evaluated their efficacy <i>in vitro</i>. Our findings demonstrate that individual tolerability is significantly influenced by the modification's sequence, pattern, and position, with limited universal principles identifiable from this dataset. The efficacy results are probably driven by the thermodynamic balance defined by a combination of parameters. The framework presented here will serve as a reference dataset to facilitate the expansion of chemical diversity in therapeutic siRNAs.

Acute and chronic neurodegenerative diseases (NDs), including multiple sclerosis (MS), Alzheimer's disease (AD), and Parkinson's disease (PD), are characterized by neurodegeneration, which is the gradual malfunction and damage of neurons and axons in the central nervous system. Improved clinical diagnostic workups and the development and tracking of successful disease-modifying treatments are made possible by detecting appropriate neurodegenerative disease (ND) biomarkers. Important biomarkers, such as Tau proteins, amyloid-β, and α-synucleins, are essential for precise identification but are often evaluated using time-consuming, expensive, and traditional techniques like polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA). Due to its exceptional selectivity and sensitivity, electrochemical biosensing has become a popular, low-cost substitute for more conventional diagnostic methods. Nanoparticles in biosensors are particularly noteworthy because they improve electron transport and aid in immobilizing biorecognition components. Conducting polymers have shown great potential in the field of electrochemical sensing. Conducting polymers have shown great potential in electrochemical sensing. Additionally, research has shown that polypyrrole, polyaniline, poly(3,4-ethylenedioxythiophene), and poly(thiophene) are often regarded as among the best conducting polymers for fabricating electrochemical sensors. Moreover, a hydrogel biosensor allows for the detection of many parameters simultaneously with real-time monitoring, allowing for more accurate and timely tracking of multiple indicators of a patient. Hydrogel nano(bio)composite sensors that use electrochemical transduction methods to detect analytes are also available. Hydrogel-based polymer sensors for early-stage neurodegenerative diagnosis are examined in this review in a novel way. Afterward, we reviewed electrochemical sensors developed for detecting biomarkers related to diseases, including multiple sclerosis, Alzheimer's, Parkinson's, and Huntington's. There have also been developments devised to enhance efficacy of electrochemical diagnostic tools to address their limitations. In this respect, we have also reviewed many polymers used in electrochemical diagnosis of neurological disorders. Finally, we have also evaluated the limits and prospects of clinical trials involving these electrochemical means of diagnoses.

The phenomenon of inflammaging, characterized by an increase in low-grade chronic inflammation, is closely associated with diseases related to liver dysfunction. This study investigated daily plasma exchange between 5-week-old and 24-month-old Sprague Dawley rats for 30 days, focusing on protein secondary structures, NLRP3 inflammasome, and necroptosis. Conformation changes in protein secondary structures were identified by infrared spectroscopy-based pattern recognition analysis. Liver biopsies with histochemical and immunohistochemical staining were used to assess molecules associated with inflammation, necroptosis and NLRP3 inflammasome complex. Expression levels of NLRP3 components were determined by qPCR. Enhanced random coils, 3₁₀ helices, β-turns, and loop structures were identified in old rats and young rats with old plasma. Young rats and old rats with young plasma displayed higher α-helices and β-sheet structures. Young rats with old plasma showed increased NLRP3, ASC, caspase-1, IL-1β, and IL-18 mRNA levels, indicating an inflammatory response. Whereas old rats with young plasma exhibited lower inflammation levels. Histological evaluations revealed that young rats receiving aged plasma showed significantly increased levels of NLRP3, ASC, caspase-1, IL-1β, TNF-α, VEGFR2, RIPK1, and MLKL immunoreactivity, whereas decreased immunoreactivity in aged rats receiving young plasma. These findings suggest that young plasma reduces NLRP3 inflammasome activation and necroptosis in aged rats.

In this experiment, the adsorption characteristics and release risk of phosphorus from three types of soils-namely no-tillage with straw off the field (CK), no-tillage with straw crushed and mulched (FS), and no-tillage with straw high-stubble mulched (GH), were investigated using a batch experimental method. The data showed that the adsorption of phosphorus was divided into three stages and reached equilibrium at 480 min. Compared with CK, FS and GH treatments reduced phosphorus adsorption by 14.68% and 24.63%, respectively. The highest adsorption occurred at pH 7, and the adsorption of phosphorus by the soil gradually decreased with increasing nitrogen concentration. Correlation analysis of soil physicochemical properties and adsorption characteristics showed that the maximum adsorption of phosphorus (Qmax) was significantly negatively correlated with the available nitrogen (AN) and positively correlated with the pH value. The environmental risk index (ERI) followed the order to GH > FS > CK, with physicochemical interactions contributing 87.13% to phosphorus adsorption. Comprehensive analysis concluded that straw return decreased the soil's phosphorus adsorption capacity, enhanced phosphorus availability, but increased the risk of phosphorus release. Appropriately increasing soil pH and reducing nitrogen fertilizer application can enhance phosphorus adsorption.

Trinucleotide repeat (TNR) diseases are neurological disorders caused by expanded genomic TNRs that become unstable in a length-dependent manner. The CAG•CTG sequence is found in approximately one-third of pathogenic TNR loci, including the HTT gene that causes Huntington's disease. Friedreich's ataxia, the most prevalent hereditary ataxia, results from GAA repeat expansion at the FXN gene. Here we used cytosine and adenine base editing to reduce the repetitiveness of TNRs in patient cells and in mice. Base editors introduced G•C>A•T and A•T>G•C interruptions at CAG and GAA repeats, mimicking stable, nonpathogenic alleles that naturally occur in people. AAV9 delivery of optimized base editors in Htt.Q111 Huntington's disease and YG8s Friedreich's ataxia mice resulted in efficient editing in transduced tissues, and significantly reduced repeat expansion in the central nervous system. These findings demonstrate that introducing interruptions in pathogenic TNRs can mitigate a key neurological feature of TNR diseases in vivo.

A growing part of the human population is affected by circadian misalignment caused by deregulated sleep, increased nighttime light exposure and erratic eating patterns. Thus, circadian rhythms are a key research area, with compelling links to cancer. Here, we review the circadian regulation of critical cellular processes, including immunity, metabolism, cell cycle control and DNA repair, under physiological homeostasis and in cancer. We discuss the divergent evidence indicating tissue-specific roles of the circadian clock in different cancer types and the potential link between circadian misalignment and early-onset cancers. Finally, we outline how understanding the circadian clock can improve cancer prevention and chronomedicine-based therapies.

<h4>Background</h4>Toxoplasma gondii can cause severe damage to immunodeficient hosts, and also compromise brain structure and function in immunocompetent hosts during latent infection. In China, the two different isolates, Chinese I (ToxoDB#9) and Chinese III are dominant epidemic strains widely spreading in humans and domestic animals and can lead to latent infection in host brain tissues, but the comparison of their manipulation patterns and mechanisms remains unclear.<h4>Methods</h4>Tachyzoites of the TgWh6 (Wh6) strain and the TgCtLHG (LHG) strain were used for establishing in vitro infection models within mouse microglia BV2 cells, and the differences in their invasion and proliferation patterns were observed. C57BL/6 J mice were used to establish in vivo latent infection models. After behavioral tests, the differential expressed transcripts (DETs) of the infected and control animals' cerebral cortex were sequenced by Nanopore RNA-seq. Functional differences of DETs were analyzed by Gene Ontology enrichment analysis (GO), Kyoto Encyclopedia of Genes and Genomes enrichment analysis (KEGG), and protein-protein interaction (PPI) and cluster analysis. Expression of the key candidates were verified by quantitative polymerase chain reaction (qPCR).<h4>Results</h4>In our infection models, we found that Wh6 had more vigorous invasion and proliferation abilities in vitro, while LHG had a greater ability to form cysts in vivo. In the latent infection phase, behavioral changes, including spatial working memory, cognitive and motor abilities, and anxiety, were observed in both Wh6 and LHG infected mice; however, the LHG group showed more serious anxiety. Among DETs, genes related to major histocompatibility complex (MHC) class II molecules were significantly upregulated in the infected mice, while genes related to synaptic transmission and neurodegenerative diseases were downregulated in the infected groups. The downregulated DETs of Sept4, Kcng4, Unc13c, and Prkcg in the WH6 group, which are related to synaptic transmission, and Ndrg2 and Arc in the LHG group, which are related to neurodegenerative diseases, were selected to be the key candidates in the latent infection phase.<h4>Conclusions</h4>Compared with WH6, although LHG has a milder invasion ability, it can cause increased behavioral disorders in hosts. Genes related to synaptic transmission and neurodegenerative diseases may be the main causes of host mental and behavioral disorders.

The diverse perspectives offered by multi-omics data analysis can aid in identifying the most relevant molecular pathways involved in disease processes, and findings in one layer can substantiate findings in other layers of information. Integrating data from multiple omics sources is becoming increasingly important to improve disease diagnosis and treatment, especially for conditions with complex and poorly understood underlying pathomechanisms. Methylmalonic aciduria (MMA), an inherited metabolic disorder, serves as an illustrative example of such a disease with poorly understood pathogenesis for which published multi-omics data are readily available. Reusing these FAIR data, obtained from the multi-omics digitization of 230 individuals (210 patients with MMA and 20 controls), we pursued advanced data integration and analysis strategies to integrate different levels of biological information, combining genomic, transcriptomic, proteomic, and metabolomic profiling with biochemical and clinical data, with the aim of elucidating molecular perturbations in individuals affected by MMA. The analysis of protein-quantitative trait loci highlighted the importance of glutathione metabolism in the pathogenesis of MMA. This finding was supported by correlation network analyses that integrated proteomics and metabolomics data, alongside gene set enrichment and transcription factor analyses based on disease severity from transcriptomic data. The correlation network analysis also revealed that lysosomal function is compromised in patients with MMA, which is critical for maintaining metabolic balance. Our research introduces a comprehensive data analysis framework that effectively addresses the challenge of prioritizing disruptions in molecular pathways by accumulating evidence from multiple omics levels.

<h4>Background</h4>Plasma-based assays or multigene panel testing can be used for the diagnosis of inherited venous thromboembolism (VTE). Our recent multigene panel data showed strictly concordant results between genetic and phenotypic laboratory testing in only half of the patients.<h4>Objectives</h4>We aimed to evaluate in detail the correlation between genotype and laboratory phenotype for defects in natural anticoagulants.<h4>Methods</h4>Gene panel test results were compared with standard thrombophilia laboratory assay data, including protein C, protein S, and antithrombin plasma activity or antigen levels. We performed additional functional protein C and protein S clot-based assays and mass spectrometry for the detection of antithrombin molecular proteoforms.<h4>Results</h4>We detected PROC, PROS1, or SERPINC1 variants in 110 of 317 patients with VTE of which 61% were (likely)pathogenic variants. Oligogenic inheritance was present in 33% of all patients. Correlation studies showed that not all variants were associated with reduced plasma activity levels, while 14%, 20%, and 5% of our patients with VTE but without a genetic variant had reduced levels for protein C, protein S, and antithrombin, respectively. Additional clot-based assays could diagnose additional patients but not all. Mass spectrometry-based antithrombin assay was able to detect SERPINC1 missense variants in plasmas that were associated with normal antithrombin activity levels.<h4>Conclusions</h4>Thrombophilia screening using plasma-based assays can lead to potential diagnostic misclassification. In contrast, multigene panel testing is more sensitive but still associated with the detection of variants of uncertain significance. Additional studies are required to clarify the role of panel testing for inherited VTE and the impact of oligogenic inheritance.

<h4>Background</h4>Cervical cancer, the fourth leading cause of female cancer mortality globally, faces treatment limitations due to drug resistance and few therapeutic options. This study seeks to identify novel therapeutic targets to address this urgent clinical need.<h4>Methods</h4>Our team identified differentially expressed genes (DEGs) in cervical cancer using gene expression omnibus (GEO) datasets. Subsequently, Mendelian randomization (MR) analysis identified causal gene-cancer relationships, followed by enrichment analysis and The Cancer Genome Atlas (TCGA) validation. Finally, we further validated the functions of the selected target genes in cervical cancer cells and analyzed their Gene Set Enrichment Analysis (GSEA) results, drug sensitivity, and prognostic value.<h4>Results</h4>We identified 2,801 upregulated and 1,646 downregulated DEGs. MR analysis identified 21 key cervical cancer-associated genes (14 upregulated, 7 downregulated), with TCGA validation confirming significant differential expression patterns. Among them, few studies have examined these core genes, particularly MERTK and SERPINF1, in cervical cancer. Experiments showed that MERTK and SERPINF1 play a role in cervical cancer. These genes help cancer cells grow, spread, and invade surrounding tissue. Mechanistically, MERTK regulates immune infiltration, whereas SERPINF1 modulates chromosomal activity. Clinically, SERPINF1 enhances overall survival (OS), disease-specific survival (DSS), and progression-free interval (PFI) in individuals with cervical cancer. Moreover, we discovered that several commonly used drugs for cervical cancer treatment, like paclitaxel, showed high efficacy against MERTK and SERPINF1.<h4>Conclusion</h4>Our study uncovers MERTK and SERPINF1 as critical regulators of cervical cancer progression and survival, offering mechanistic insights into their roles in tumor behavior and the immune microenvironment. These findings provide a foundation for precision therapies, with SERPINF1 restoration and MERTK inhibition as promising strategies. Clinical translation of these targets could address current treatment limitations.

<h4>Objective</h4>This study aims to elucidate the role of aryl hydrocarbon receptor nuclear transporter 2 (ARNT2) in cervical cancer (CC) and explore the potential mechanism by which ARNT2 promotes the progression of CC through the protein phosphatase 2A (PP2A)/Akt signaling pathway.<h4>Methods</h4>Bioinformatics tools were used to analyze the expression level of ARNT2 in cancer and its correlation with cancer prognosis. Western Blot and immunohistochemistry staining were used to detect the expression of ARNT2 protein in CC tissues and cells. ARNT2 was knocked down in SiHa and HeLa cells, respectively. Cell Counting Kit-8 assay and colony formation assay were used to detect changes in cell proliferation. Transwell assay and plate scratch assay were used to detect changes in cell migration and invasion. Western Blot assay was used to detect changes in the expression of PP2A/Akt signaling pathway after ARNT2 expression was downregulated. Finally, a CC xenograft tumor model was constructed to evaluate the effect of ARNT2 on SiHa cell tumorigenesis in vivo.<h4>Results</h4>ARNT2 is highly expressed in tumor tissues and cell lines. ARNT2 knockdown can significantly inhibit the proliferation, invasion and migration of SiHa and HeLa cells in vitro and in xenograft models. Further studies have shown that ARNT2 may promote tumor formation by regulating the PP2A/Akt pathway.<h4>Conclusion</h4>ARNT2 promotes the malignant biological behavior of CC cells through the PP2A/Akt signaling pathway, confirming its potential as a prognostic marker for CC.

<h4>Introduction</h4>Cardioplexol™ was recently proven effective and non-inferior to Buckberg's solution in a pivotal Phase-3 clinical trial. We hypothesized here that a standardized training program for surgeons without prior experience of Cardioplexol™ could increase its administration reliability and participate to its overall benefit.<h4>Methods</h4>Open label, single group, observational study involving 29 surgeons from 7 centers in 3 countries. The training program included a theoretical part, and two surgical procedures performed under trainer supervision. In a subsequent evaluation part, surgeons operated on 4 additional patients. The number of major deviations from the pre-defined administration protocol (incorrect volume of initial/second/third/fourth dose, incorrect duration of injection of initial dose, incorrect timing of application of initial/second/third/fourth dose) was set as primary endpoint.<h4>Results</h4>A total of 171 patients were screened of which 157 were operated on (57 in the training part and 100 in the evaluation part). No major deviations were observed. Other outcomes, including postoperative TnT and CK-MB profiles, cumulative inotropic support provided during the first 24 h after myocardial reperfusion, cardiac conversion rate, ICU length of stay, were all similar to or better than the results observed in the previous pivotal study.<h4>Conclusion</h4>Cardiac surgeons not familiar to Cardioplexol™ benefit from a structured and supervised training. This kind of training contributes to improve the efficiency and safety of a new cardioplegic solution such as Cardioplexol™.<h4>Trial registration</h4>[ClinicalTrials.gov]: identifier [NCT03823521, and EudraCT No: 2018-002311-10].

<h4>Background</h4>Colorectal cancer (CRC) ranks among the most prevalent malignancies of the gastrointestinal tract and remains a leading cause of cancer-related mortality worldwide. Although telomere biology has been increasingly implicated in immune modulation and tumor progression, its clinical significance in CRC remains poorly understood.<h4>Methods</h4>We developed a telomere score, termed TELscore, by integrating transcriptomic and intratumoral microbiome profiles from publicly available colorectal cancer (CRC) cohorts. To comprehensively characterize TELscore subgroups, we performed pathway enrichment analysis, tumor immune microenvironment (TIME) profiling, and microbiome niche assessment. Whole-slide histopathological images (WSIs) and immunohistochemical (IHC) staining were utilized to visualize immune features, including tertiary lymphoid structures (TLSs), across subgroups. Patients were stratified into high and low TELscore categories, and the predictive robustness was validated across multiple independent training and validation cohorts. Chemotherapeutic drug sensitivity was evaluated using pharmacogenomic data from the Genomics of Drug Sensitivity in Cancer (GDSC) database. Furthermore, the predictive capacity of TELscore for immunotherapy response was independently assessed in an external cohort. Finally, single-cell RNA sequencing (scRNA-seq) analysis was conducted to further dissect the cellular landscape and immunological heterogeneity within the TME.<h4>Results</h4>TELscore stratified patients into two biologically and clinically distinct subgroups. The high TELscore group, which exhibited significantly shorter DFS, showed marked enrichment of tumorigenic pathways such as EMT, along with a distinctly immunosuppressive TME. This was reflected by elevated ESTIMATE/TIDE scores and corroborated by CIBERSORT, which revealed increased infiltration of M0 macrophages and upregulation of immunosuppressive signatures. In contrast, the low TELscore group was enriched for cell cycle related pathways, including E2F targets and the G2/M checkpoint, and demonstrated higher infiltration of pro-inflammatory M1 macrophages. 16S rRNA sequencing further revealed a divergent intratumoral microbiome between subgroups, the high TELscore group harbored significantly greater relative abundance of Selenomonas and Lachnoclostridium, two pathogenic genera previously associated with colorectal tumorigenesis. Complementary histopathological assessment via WSI demonstrated a marked absence of intraTLSs in high TELscore tumors. From a therapeutic standpoint, high TELscore tumors exhibited reduced sensitivity to standard chemotherapeutic agents-including Fluorouracil, Irinotecan, Oxaliplatin, and Docetaxel-as reflected by elevated IC50 values. Conversely, these tumors demonstrated increased susceptibility to MAPK pathway inhibitors, such as Selumetinib and Trametinib. Notably, TELscore also served as a robust predictor of immunotherapy response, which was validated in the IMvigor210 cohort. Finally, scRNA analysis highlighted profound cellular and functional divergence between TELscore subgroups. We identified intensified intercellular communication between inflammatory macrophages and fibroblasts, reinforcing the presence of an immunosuppressive niche.<h4>Conclusion</h4>TELscore is a robust stratification tool that captures the interplay between tumor biology, immune characteristics, and microbial ecology in colorectal cancer. By identifying clinically relevant subtypes with distinct therapeutic vulnerabilities, TELscore offers a powerful framework to advance personalized treatment and precision oncology.

Cardiovascular diseases (CVDs) remain the leading cause of death worldwide, both in developed and developing countries. Despite the implementation of various measures in clinical practice that have shown certain curative effects, poor prognosis and irreversible pathological cardiac remodeling continue to limit the therapeutic effect of CVDs. There are still many new mechanisms worth exploring for the regulation of CVDs. Previous studies have highlighted the potential applicability of exosomes in CVDs, and significant research has been conducted in this area. In this review, we summarize the physiological mechanisms of exosomes and the basic research achievements in regulating CVDs via exosomal non-coding RNAs. We also discuss the limitations and prospects of exosome application in CVD treatment.

<h4>Background</h4>Huntington's disease (HD) is an inherited, neurodegenerative disorder caused by the expansion of an unstable CAG repeat sequence in the Huntingtin (HTT) gene. The prevalence of HD, allelic diversity, rate of novel expansions and the clinical correlates vary across populations.<h4>Objective</h4>We analyzed the diversity of alleles and their clinical correlates and examined the inheritance patterns and the pattern of instability of CAG repeats in a few families.<h4>Methods</h4>Clinical history and pedigree structure were collected from records or through interviews between 2016 and 2019. Genetic testing at the HD locus was done on clinical suspicion, or relatedness, after counseling. Descriptive statistics and correlation analysis were used.<h4>Results</h4>Expanded repeats were detected in 239 individuals, including 232 who were symptomatic and 7 presymptomatic relatives. The number of CAG repeats (mean = 45.6) and age at onset (mean = 39.2 years) showed a strong inverse correlation (<i>r</i> = -0.67). We found atypical alleles such as 8 intermediate alleles (IA), 12 reduced penetrance alleles and 14 large (>60) expansion alleles corresponding to juvenile HD. Three individuals carried biallelic expansions. Paternal inheritance was more common, and the mean increase in repeats in the available parent-child pairs was 14. Thirty-seven individuals had no family history of HD, with <i>de novo</i> expansion confirmed in three cases.<h4>Conclusions</h4>Novel mutations at the HTT locus may not be rare in India. A lack of family history should not exclude appropriate testing. The prevalence of IA and incidence of <i>de novo</i> expansions suggest that there may be a reservoir of alleles prone to expansion.

In this review, we discuss how mutations in glioma are associated with prognosis and treatment efficacy. A fascinating characteristic of glioma and all cancers is that while common growth and developmental pathways are altered, the characteristic mutations are distinct depending on the specific type of tumor with concomitant prognoses. Next-generation sequencing, precision medicine, and artificial intelligence are boosting the employment of molecular biomarkers in cancer diagnosis and treatment. Understanding the biological underpinnings of distinct mutations on critical signaling pathways is crucial for developing novel therapies for glioma.

Increases in flow elicit dilations in the basilar artery (BA) supplied by the posterior cerebral circulation (PCC), and ensuring efficient blood supply to the circle of Willis in which blood flow and pressure can distribute and equalize, and thus provide the appropriate supply for the daughter branches to reach certain brain areas. In contrast, increases in flow elicit constrictions in the middle cerebral artery (MCA), supplied by the anterior cerebral circulation (ACC) and regulating the blood pressure and flow in distal cerebral circulation. Mediators of flow-dependent responses include arachidonic acid (AA) metabolites and nitric oxide (NO). We hypothesized that mediators of flow-dependent responses are differentially expressed in cerebral arteries of the PCC (CA_PCC) and ACC (CA_ACC). The expressions of key enzymes of the AA pathway-cyclooxygenases (COX1/COX2), cytochrome P450 hydroxylases (Cyp450), thromboxane synthase (TXAS), thromboxane A2 (TP) receptor, prostacyclin synthase (PGIS), prostacyclin (IP) receptor (IP); neuronal nitric oxide synthase (nNOS), and endothelial nitric oxide synthase (eNOS)-in the BA and MCA from rats (n = 20) were determined by western blotting. Transcriptome analysis in CA_PCC and CA_ACC from rats (n = 25) was assessed by RNA sequencing. In BA compared to MCA, COX1/2 and Cyp450 protein expressions were lower, PGIS was higher, TXAS and nNOS/eNOS were similar, TP receptors were lower, and IP receptors were higher. Gene expressions of vasodilator canonical pathways were higher in CA_PCC; vasoconstriction canonical pathways were higher in CA_ACC. Mediators of flow-dependent vasomotor signaling are differentially expressed in cerebral arteries of the posterior and anterior circulation, corresponding to their vasomotor function.

Hereditary hemochromatosis is a genetic disorder caused by mutations in genes regulating hepcidin, leading to increased iron absorption and progressive accumulation in the body. It is commonly encountered in the Western population, but prevalence in the Indian population, especially of the <i>H63D</i> homozygous mutation variant, is on the lower end. Herein is a case of a young male with a paternal hereditary trait of polydactyly, on regular follow-up for anti-hypertensives, who presented with fatigue and multiple large joint pains. Unexplained fatigue, along with a significant family history, warranted further evaluation. Serum iron levels, transferrin saturation, and ferritin levels were elevated. Hence, the patient was referred to the hematology department, for which <i>HFE </i>genetic mutation analysis was done, and<i> a</i> <i>H63D</i> homozygous mutant was noted. Target end-organ damage was assessed. The patient was monitored and advised to reduce iron intake. Genetic counseling was also done, along with screening of first- and second-degree relatives.

Background Iron deficiency anemia (IDA) is a highly prevalent disorder among the pediatric population and has a significant impact on growth and health. Ferrous sulfate is commonly prescribed for the treatment of IDA; however, its efficacy in the pediatric population requires further research. Objective This study aimed to evaluate the efficacy of ferrous sulfate in improving hemoglobin (Hb) and ferritin levels in children diagnosed with IDA. Methods A quasi-experimental study was conducted involving 108 participants aged 1-8 years who were diagnosed with IDA. Participants received ferrous sulfate supplements for three months. Hemoglobin and serum ferritin levels were measured both at baseline and after three months of treatment.  Results Ferrous sulfate administration resulted in a significant increase in hemoglobin levels by an average of 3.11 mg/dl and a net increase in ferritin levels by 5.9 ng/ml after 3 months of treatment. Additionally, a positive association was found between low Hb levels and underweight children<i>.</i> Conclusion Ferrous sulfate is an effective therapeutic agent for managing IDA in children, leading to significant improvements in hemoglobin and ferritin levels. Further research involving RCTs with longer follow-up periods is recommended.

<i>Background and Objectives</i>: Chromobox 1 (CBX1), a key epigenetic regulator involved in chromatin remodeling, has been implicated in various cancers; however, its role in liver hepatocellular carcinoma (LIHC) remains underexplored. This study aimed to investigate the expression patterns, epigenetic regulation, and non-coding RNA (ncRNA) networks involving CBX1 in LIHC, assess their potential as diagnostic and prognostic biomarkers, and explore their relevance as a putative therapeutic target. <i>Materials and Methods</i>: A multi-omics bioinformatics approach was employed using datasets from GEPIA2, OncoDB, UALCAN, Human Protein Atlas, KM Plotter, MethSurv, miRNet, and ENCORI. These databases were used to analyze mRNA and protein expression, DNA methylation, prognosis, and interaction networks involving CBX1 and ncRNAs. <i>Results</i>: CBX1 was significantly upregulated in both the mRNA and protein expression in LIHC. Upregulated CBX1 expression was associated with poor prognosis. DNA methylation analysis revealed that both hypermethylated and hypomethylated probes were significantly associated with CBX1 expression and poor prognosis. hsa-miR-212-3p and hsa-miR-132-3p were significantly upregulated in LIHC and were positively correlated with CBX1 expression and poor prognosis. The ncRNA network was identified, including long ncRNAs, circular RNAs, and pseudogenes, many of which were linked to tumor progression and poor prognosis, and competing endogenous RNAs were associated with tumor progression and poor prognosis in LIHC. <i>Conclusions</i>: CBX1 was significantly overexpressed in LIHC and was regulated by both DNA methylation and ncRNA interactions. Its expression is closely associated with a poor prognosis. The CBX1-micro-RNA-long ncRNA/circular RNA axis is a promising avenue for the development of novel diagnostic and therapeutic strategies. This study provides system-level insights into the regulatory landscape of CBX1 in LIHC and supports its potential role in precision medicine.

Huntington&#039;s Disease (HD) originates from the expansion of a polyglutamine (PolyQ) tract in the Huntingtin protein (Htt), which can assume a Coiled-coil fold (Cc). We previously found that Cc structures mediate the aggregation and toxicity of polyQ Htt. Since polyQ Htt aggregates were previously found to be internalized by cells, here we hypothesize that Cc structures might be implicated in the intercellular propagation of Htt aggregates. To test this hypothesis, we performed experiments using human cell lines expressing Htt proteins with different probabilities to acquire a Cc fold. We found that Htt with reduced Cc structures were released significantly less compared to Htt with intact Cc structures. We also found that Cc structures mediate the internalization of Htt proteins in recipient cells. Together, these results underlie the importance of the Cc structure in the process of intercellular propagation of Htt polyQ aggregates and suggest that interfering with Cc formation might be a therapeutic strategy for HD.

<h4>Introduction</h4>Over the last decade, immune checkpoint inhibitors (ICIs) like PD-1/PD-L1 or CTLA-4, which reinvigorate T cells for tumor control have become standard-of-care treatment options. In response to the increasingly recognized mechanisms of resistance to T cell activation in immunologically cold tumors, immuno-oncology drug development has started to shift beyond T cell approaches. These include tumor-associated macrophages (TAMs), a major pro-tumor immune cell population in the tumor microenvironment known to silence immune responses.<h4>Areas covered</h4>Here we outline anti-TAM therapies in current development, either as monotherapy or in combination with other treatment modalities. We describe emerging drugs targeting TAMs under investigation in phase II and III testing with a focus on their distinguishing mechanism of action which include (1) reprogramming of TAMs toward anti-tumor function and immune surveillance, (2) blockade of recruitment, and (3) reduction and ablation of TAMs.<h4>Expert opinion</h4>Several new immuno-oncology agents are under investigation to harness anti-tumor functions of TAMs. While robust anti-tumor efficacy of anti-TAM therapies across advanced solid organ cancers remains elusive to-date, TAM reprogramming therapies have yielded benefits in select cancers. The inherent heterogeneity of the diverse TAM population will require enhanced investments into biomarker-driven approaches to fully leverage its therapeutic potential.

Cervical cancer (CC) remains a significant global health concern, particularly affecting women in low-income countries. Despite advancements in screening programs, CC continues to pose a substantial mortality risk, highlighting the need to explore diagnostic and treatment modalities. This review focuses on the role of deregulated microRNAs (miRNAs) in CC development, emphasizing their potential as biomarkers for early detection and prognosis in body fluids. miRNAs have emerged as critical regulators of key cellular processes, including proliferation, migration, invasion, and apoptosis, and their dysregulation is closely linked to CC progression. Upregulated miRNAs such as miR-146b-3p, miR-1908, and miR-21 promote CC progression by targeting tumor suppressor genes, while downregulated miRNAs like miR-23-3p and miR-4262 are associated with reduced tumor aggressiveness. miRNAs also hold significant promise as non-invasive prognostic biomarkers. Their expression levels correlate with clinical outcomes, including tumor stage, metastasis, and overall survival, making them valuable tools for risk stratification and personalized treatment strategies. Liquid biopsies, which detect circulating miRNAs in bodily fluids, offer a minimally invasive approach to monitor tumor dynamics and predict patient outcomes. Furthermore, exosomal miRNAs are emerging as promising diagnostic and prognostic tools for CC. Advanced diagnostic technologies and bioinformatics tools are anticipated to enhance the identification of evident miRNA biomarkers in the clinical settings. Standardized protocols for sample collection and analysis will improve the reproducibility of miRNA studies, while a deeper understanding of miRNA biology may unlock their potential as therapeutic targets. In conclusion, this review consolidates current research on deregulated miRNAs in CC, highlighting their diagnostic and prognostic significance. The findings underscore the potential of miRNAs to revolutionize CC management through innovative diagnostic and therapeutic strategies.

Amoxicillin is one of the most widely used antibacterial drugs. It is known to be the first reason of adverse effects caused by medications in human and animals. Antibacterial drugs, combined with a delivery agent, could provide desirable therapeutic effects with decreased toxicity and reduce the emergence of antimicrobial resistant bacteria. The aim of this study was to compare the toxicity of amoxicillin, polyphosphate ester amoxicillin complex and phosphate ester on mice. Synthesis of the polyphosphate ester (P4) was performed via polycondensation technique, using PEG-400. Animal studies were performed in accordance with the European Convention for the Protection of Vertebrate Animals (Strasbourg, 1986). BALB/c mice were treated by intramuscular injection with saline 0.2 mL (control), amoxicillin 0.2 mL (15 mg/kg); polyphosphate ester complex with amoxicillin 0.2 mL (with amoxicillin content 15 mg/kg) and polyphosphate ester 0.2 mL. Blood biochemical analysis and histology of liver, spleen and kidney were used to assess toxicity. Blood biochemical analysis indicates that P4 did not induce changes in liver and kidneys. Specifically, blood biochemical indicis that represent functional state and cell structure of these organs were within normal physiological values: ALT (56 ± 15.96 U/L), AST (265 ± 37.50U/L), urea (4.4 ± 1.45 mmol/L), creatinine 62.8 ± 5.17 mmol/L, cholesterol 3.5 ± 0.56 mmol/L, total protein 55.9 ± 4.60 g/L, glucose 8.1 ± 0.55 mmol/L. However, the analysis of organ to body weight ratio showed decreased liver ratio (p ≤ 0.05) in mice injected with polyphosphate ester (P4). Histological examination of the liver didn't show severe pathological changes. There were single places with mild portal vein inflammation in liver of mice receiving amoxicillin and amoxicillin complexed with polyphosphate ester. P4 separately in some places caused cell cytoplasm granulation in liver. No spleen alterations were observed. Overall, the results of this study showed that P4 polyphosphate ester alone and in complex with amoxicillin does not cause renal, hepatic and splenic toxicity in mice. Thus, polyphosphate ester P4 can serve as a safe drug carrier for antimicrobial drugs. It is planned to carry out more extensive studies on other animal species to study its biocompatibility and effectiveness of antimicrobial activity in a complex with antimicrobials.

This open-label, single-arm phase II study assessed the safety and efficacy of sequential hypofractionated radiotherapy (RT) followed by zimberelimab and R-GemOx (rituximab, gemcitabine, oxaliplatin) in patients with primary refractory diffuse large B-cell lymphoma (DLBCL). Fourteen patients were enrolled between June 2022 and December 2023, with 13 included in the analysis. RT doses of 36 and 24 Gy were delivered to the gross and target volumes in 12 fractions, followed by zimberelimab and R-GemOx. The overall response rate within the irradiated field was 92.3%, and a complete response (CR) was achieved by 61.5% of patients; however, 38.5% experienced disease progression. Treatment-related toxicities were manageable, primarily comprising mild leukocytopenia. Digital spatial profiling revealed 53 differentially expressed genes in CD20-rich lymphoma regions and 93 in CD3-rich T cell regions in non-CR patients. Reactome analysis identified key immune system pathways. T cell infiltration correlated with treatment efficacy, and multiplex immunohistochemistry validated immune pathways as potential therapeutic targets. This study demonstrated the promising role of RT combined with immunochemotherapy in refractory DLBCL and suggests immune pathways as critical targets to improve treatment outcomes.

Axon guidance, a fundamental process in neural circuit formation, is intricately regulated by Fibroblast Growth Factors (FGFs) and their receptors (FGFRs) through dynamic cytoskeletal remodeling. FGF signaling, mediated by heparan sulfate proteoglycans or Klotho co-factors, activates key downstream pathways: PI3K-Akt, JAK-STAT, PLCγ, and RAS-MAPK. These pathways orchestrate actin filament dynamics, microtubule stability, and the organization of intermediate filaments. These pathways converge on Rho GTPases, cofilin, profilin, and tau to balance the cytoskeletal assembly-disassembly cycles, enabling growth cone navigation. Unresolved questions, such as the mechanisms underlying FGF-mediated growth cone steering, highlight critical future research directions. This review integrates structural, molecular, and functional insights into how FGF-FGFR interactions regulate axon pathfinding, emphasizing the crosstalk between signaling cascades and cytoskeletal plasticity. Elucidating these mechanisms not only advances our understanding of neural development but also opens therapeutic avenues for neuro-developmental disorders, nerve injury, and neurodegenerative diseases by targeting FGF-driven cytoskeletal dynamics.

Boys' pubertal overweight associates with future offspring's asthma and low lung function. To identify how paternal overweight is associated with offspring's DNA methylation (DNAm), we conducted an epigenome-wide association study of father's body silhouette (FBS) at three timepoints (age 8, voice break and 30) and change in FBS between these times, with offspring DNAm, in the RHINESSA cohort (N = 339). We identified 2005 differentially methylated cytosine-phosphate-guanine (dmCpG) sites (FDR < 0.05), including dmCpGs associated with offspring asthma (119), lung function (178) and BMI (291). Voice break FBS associated with dmCpGs in loci including KCNJ10, FERMT1, NCK2 and WWP1. Change in FBS across sexual maturation associated with DNAm at loci including NOP10, TRRAP, EFHD1, MRPL17 and NORD59A;ATP5B and showed strong correlation in reduced gene expression in loci NAP1L5, ATP5B, ZNF695, ZNF600, VTRNA2-1, SOAT2 and AGPAT2. We identified 24 imprinted genes including: VTRNA2-1, BLCAP, WT1, NAP1L5 and PTPRN2. Identified pathways relate to lipid and glucose metabolism and adipogenesis. Father's overweight at puberty and during reproductive maturation was strongly associated with offspring DNA, suggesting a key role for epigenetic mechanisms in intergenerational transfer from father to offspring in humans. The results support an important vulnerability window in male puberty for future offspring health.

<h4>Background</h4>The interplay between diet and healthspan is a topic of great interest in biomedical research. Toward this end, consumption of marine omega-3 fatty acids is of particular significance, as reports suggest that diets focused on seafood can prolong the disease-free portion of the human lifespan. Fish consumption has also been linked to reduced biological aging as measured by epigenetic clocks, but there is no conclusive evidence of a causal relationship. Moreover, fish oils reduce triglycerides, and may affect other lipid profiles, as well as systemic inflammation. To investigate further, we used two-sample Mendelian randomization to investigate potential causality between fish intake and healthspan markers.<h4>Methods</h4>Bidirectional Mendelian randomization was performed in the two-sample setting with publicly available GWAS summary statistics. GWAS data from the UK Biobank for oily fish consumption (n = 460,443) and fish oil supplementation (n = 461,384) were used as the primary exposures. First-generation epigenetic clocks Hannum age and intrinsic epigenetic age acceleration (IEAA), as well as second-generation clocks GrimAge and PhenoAge were collected from an independent dataset of individuals of European ancestry (n = [34,449-34,667]). Finally, data from the Integrative Epidemiology Unit database was used for serum proxies of lipidemia and systemic inflammation (n = [61,308-78,700]). Additional sensitivity analyses, such as reverse causation testing and the Cochran's Q test were performed for exposure-outcome pairs where the inverse variance weighted (IVW) method was significant (p-value < 0.05), and where the MR Egger method indicated an effect in the same direction as the IVW result.<h4>Results</h4>We report that oily fish consumption appears to decrease PhenoAge acceleration (p < 0.0086), whereas fish oil supplementation appears to decrease GrimAge (p = 0.037). Both omega-3 exposures modify the epigenetic clocks in the expected negative, or age-decelerating, direction. For the serum biomarkers, we find evidence that fish oil consumption leads to a reduction in triglycerides (p = 0.004), although HDL and LDL were not significantly modified. Finally, we also detected a suggestive inverse relationship between oily fish consumption and hsCRP (p = 0.064).<h4>Conclusions</h4>Our analysis shows that consuming fish oil, whether through whole food or as a supplement, can have a rejuvenating impact as measured by PhenoAge and GrimAge acceleration. We have also provided evidence further linking fish oil intake and lower triglyceride levels. These results, based on robust MR-based analyses, emphasize the effectiveness of dietary choices in modifying emerging measures of healthspan.

Castrate-resistant prostate cancer (CRPC) is a likely outcome of hormone treatment for advanced prostate cancer. Although no longer dependent on androgen levels, CRPC remains driven by the androgen receptor (AR). One proposed progression mechanism is altered repertoires of coregulator proteins possessing the ability to alter AR activity. Increased expression of tripartite motif-containing 24 (TRIM24) and TRIM28-two members of a distinct bromodomain-containing subfamily of Tripartite motif (TRIM) coregulators-occurs in CRPC. Endogenous TRIM24 and TRIM28 interact with each other and AR, bind to chromatin and regulate genes such as the angiogenic factor vascular endothelial growth factor A (VEGFA) and oncogene MYC. Silencing of TRIM24 and TRIM28 simultaneously, but not either alone, sensitised CRPC model cell lines to the antiandrogen enzalutamide and bicalutamide. This re-sensitisation to antiandrogen therapeutics could then be reversed by addition of VEGF. Furthermore, both TRIM24 and TRIM28 expression associated with angiogenesis signatures in tumour samples, and conditioned media from TRIM24 and TRIM28-silenced cancer cells inhibited endothelial cell proliferation and formation of vascular tube structures. Our data suggest that TRIM24 and TRIM28 proteins interact, in gene-specific manners, to regulate AR activity, increase VEGF signalling and angiogenesis, and that targeting these coregulators may increase the effectiveness of antiandrogen therapy.

Numerous long noncoding RNAs (lncRNAs) are generated in response to external stimuli, but the scope and functions of such activity are not known. Here, this study provides insight into how the transcription of lncRNAs is connected to DNA damage response by identifying the lncRNA ZFAS1, which is required for cell cycle arrest, transcription regulation, and DNA repair. Mechanistically, ZFAS1 facilitates dynamic changes in hyperphosphorylated forms of the large subunit of RNA polymerase II (RNAPII) around transcription initiation sites by directly targeting the regulated genes. It is shown that extensive transcription shutdown and concomitant stimulated engagement of RNAPII-Ser2P are crucial for repair and cell survival upon genotoxic stress. Finally, ZFAS1 knockout in mice dampened nucleotide excision repair (NER) and led to kidney dysplasia. Overall, the findings extend the understanding of lncRNAs in DNA damage repair (DDR) and imply a protective role of lncRNA against DDR-deficient developmental disorders.

The complexity and precision of the human nervous system have posed significant challenges for researchers seeking suitable models to elucidate refractory neural disorders. Traditional approaches, including monolayer cell cultures and animal models, often fail to replicate the intricacies of human neural tissue. The advent of organoid technology derived from stem cells has addressed many of these limitations, providing highly representative platforms for studying the structure and function of the human embryonic brain and spinal cord. Researchers have induced neural organoids with regional characteristics by mimicking morphogen gradients in neural development. Recent advancements have demonstrated the utility of neural organoids in disease modeling, offering insights into the pathophysiology of various neural disorders, as well as in the field of neural regeneration. Developmental defects in neural organoids due to the lack of microglia or vascular systems are addressed. In addition to induction methods, microfluidics is used to simulate the dynamic physiological environment; bio-manufacturing technologies are employed to regulate physical signaling and shape the structure of complex organs. These technologies further expand the construction strategies and application scope of neural organoids. With the emergence of new material paradigms and advances in AI, new possibilities in the realm of neural organoids are witnessed.

PIWI-interacting RNAs (piRNAs), a novel category of small non-coding RNAs, are widely expressed in eukaryotes and deregulated in several pathologies, including cancer. Little is known about their function and mechanism in renal cell carcinoma (RCC) progression. Herein, a down-regulated piRNA in RCC, termed piR-hsa-28489 (designated as piR-RCC), is identified to impede RCC progression both in vivo and in vitro. Mechanistically, piR-RCC directly interacts with Y-box binding protein 1 (YBX-1), thus impeding p-AKT-mediated YBX-1 phosphorylation and its subsequent nuclear translocation. Moreover, YBX-1 coordinates the transcription of ETS homologous factor (EHF) as a repressor factor. Consequently, piR-RCC enhances EHF expression, leading to the inhibition of RCC proliferation and metastasis. Based on these, a biomimetic nanoparticle platform is constructed to achieve RCC-specific targeted delivery of piR-RCC. The nanoparticles are fabricated using a cell membrane coating derived from cancer cells and used to encapsulate and deliver piR-RCC plasmids to renal orthotopic implantation in mice, hindering RCC progression. This study illustrates piR-RCC/YBX-1/EHF signaling axis in RCC, offering a promising therapeutic avenue for RCC.

Small alterations during the early stages of the innate immune response to an implant can drive large changes in adaptive immunity. Biomaterials for regenerative purposes can be engineered to modulate this immune response in beneficial ways. This study presents an innovative patch designed and functionalized to target the innate immunity at the implant site. Mannose moieties are incorporated into collagen patches, resulting in a technology called Local Immunotuning Patch (LIP), designed to directly interact with antigen presenting cells through their mannose receptor. In vitro, LIP shows anti-inflammatory effects on bone marrow-derived macrophages and inhibitory properties even on methicillin-resistant bacterial strains. Subcutaneous implantation in mice reveals that LIP modulates multiple pathways related to innate and adaptive immunity, underscoring its role in shaping an immune-engineered environment around the implant. These findings highlight the potential of this strategy to control the foreign body reaction at the implant site, making it applicable for various uses, including wound healing and surgical infection control in reconstructive procedures.

<h4>Purpose of review</h4>Genome-wide association studies (GWAS) have significantly advanced osteoporosis research by identifying genetic loci associated with bone mineral density (BMD) and fracture risk. However, disparities persist due to the underrepresentation of non-European populations, limiting the applicability of polygenic risk scores (PRS). This review examines recent advancements in osteoporosis genetics, highlights existing disparities, and explores strategies for more inclusive research.<h4>Recent findings</h4>European-focused GWAS have identified key loci for osteoporosis, including WNT signaling (SOST, LRP5) and RUNX2 transcriptional regulation. However, fewer than 40% of these variants can be replicated in Asian and African populations. Emerging studies in non-European groups reveal population-specific loci, sex-specific associations, and gene-environment interactions. Advances in machine learning (ML)-assisted GWAS and multi-omics integration are improving genetic discovery. Expanding GWAS in diverse populations, integrating multi-omics data, refining ML-based risk models, and standardizing biobank data are essential for equitable osteoporosis research. Future efforts must prioritize clinical translation to enhance personalized osteoporosis prevention and treatment.

<h4>Background</h4>Pancreatic cancer is a highly aggressive cancer characterized by low survival rate. Enhanced ribosome biogenesis may be associated with tumor drug resistance and malignant phenotypes, representing a potential therapeutic target in pancreatic cancer. Therefore, exploring the molecular mechanisms of ribosome biogenesis in pancreatic cancer may uncover new biomarkers and potential therapeutic targets, facilitating the development of personalized treatment strategies.<h4>Methods</h4>Ribosome biogenesis-related gene signatures were acquired from TCGA and Gene Cards databases. Prognostic gene sets were screened using machine learning algorithms to construct a risk model, which was externally validated via GEO database. Single-cell RNA sequencing analysis (GSE155698 dataset) was performed to assess gene expression patterns and module scores.<h4>Results</h4>Sixty ribosome biogenesis-related prognostic genes were identified in pancreatic cancer. Cox regression and machine learning algorithms selected nine pivotal biomarkers (ECT2; CKB; HMGA2; TPX2; ERBB3; SLC2A1; KRT13; PRSS3; CRABP2) with high diagnostic and prognostic specificity for PAAD. The machine learning-derived risk score correlated strongly with tumor proliferation pathways and immunosuppression, suggesting dual roles in tumor promotion and immunosuppressive microenvironment remodeling. Single-cell analysis highlighted predominant expression of CKB, SLC2A1, ERBB3, CRABP2, and PRSS3 in pancreatic ductal epithelial cells.<h4>Conclusions</h4>Our results shed light on the potential connections between ribosome biogenesis-related molecular characteristics and clinical features, the tumor microenvironment, and clinical drug responses. The research underscores the critical role of ribosome biogenesis in the progression and treatment resistance of pancreatic cancer, offering valuable new perspectives for prognostic evaluation and therapeutic response prediction in pancreatic cancer.

Childhood cancer survivors (CCS) often suffer from cardiac disease (CD) after treatment that included anthracycline and radiotherapy involving the heart. However, the variability in CD occurrence cannot be explained solely by these treatments, suggesting the existence of genetic predisposition. We conducted a systematic review searching on Medline-PubMed and Scopus, to identify studies reporting associations between genetic factors and CD in CCS. We included studies published up to 11 April 2023, with no lower limit, and assessed the quality of genetic associations by the Q-genie tool. As a result, 20 studies were included (15 case-control and five cohorts), revealing several genes and variants associated with cardiomyopathy, among which, SLC28A3-rs7853758, RARG-rs2229774, P2RX7-rs208294 and P2RX7-rs3751143 variants gave the most consistent findings. This review highlights the necessity to establish a set of clinically useful genes and variants to identify patients most at risk of developing cardiomyopathy, and to implement monitoring and prevention strategies.

<h4>Main conclusion</h4>This study showed that MYC2 transcriptionally regulates valuable metabolites in Taraxacum spp. through direct interaction with specific target gene promoters. The Russian dandelion (Taraxacum kok-saghyz) represents a promising alternative species, capable of producing several high-added-value compounds, including natural rubber. Nevertheless, further enhancements are required for its optimal utilization by the industry. Here, we explored the role of the bHLH transcription factor TksMYC2, homolog of AtMYC2, in the regulation of the biosynthesis of specialized metabolites and free fatty acids and in the control of natural rubber production. Metabolic analyses of Taraxacum kok-saghyz plants showed that the overexpression of TksMYC2 significantly affected the accumulation of metabolites in roots and leaves, such as sesquiterpene lactones, phenylpropanoids, and free fatty acids. Moreover, overexpressing plants presented a significant increase in natural rubber production in both Taraxacum kok-saghyz and its related species Taraxacum brevicorniculatum. The direct interaction of TksMYC2 with the regulatory regions of cis-prenyltransferase 2 (CPT2), small rubber particle proteins (SRPP1, SRPP3, and SRPP4), involved in the biosynthesis of natural rubber, and with the germacrene A oxidase (GAO), involved in the biosynthesis of sesquiterpenes, was demonstrated by chromatin immunoprecipitation coupled with quantitative PCR. Additionally, these genes were highly induced in the lines overexpressing TksMYC2. Our findings suggest that TksMYC2 and its downstream components may be valid targets for breeding programmes to increase the production of valuable metabolites, including natural rubber.

Cancer remains a significant global health burden due to its high morbidity and mortality. Oncogene-targeted therapy and immunotherapy have markedly improved the 5-year survival rate in the patients with advanced or metastatic tumors compared to outcomes in the era of chemotherapy/radiation. Nevertheless, the majority of patients remain incurable. Initial therapies eliminate the bulk of tumor cells, yet residual populations termed drug-tolerant persister cells (DTPs) survive, regenerate tumor and even drive distant metastases. Notably, DTPs frequently render tumor cross-resistance, a detrimental phenomenon observed in the patients with suboptimal responses to subsequent therapies. Analogous to species evolution, DTPs emerge as adaptative products at the cellular level, instigated by integrated intracellular stress responses to therapeutic pressures. These cells exhibit profound heterogeneity and adaptability shaped by the intricate feedforward loops among tumor cells, surrounding microenvironments and host ecology, which vary across tumor types and therapeutic regimens. In this review, we revisit the concept of DTPs, with a focus on their generation process upon targeted therapy or immunotherapy. We dissect the critical phenotypes and molecule mechanisms underlying DTPs to therapy from multiple aspects, including intracellular events, intercellular crosstalk and the distant ecologic pre-metastatic niches. We further spotlight therapeutic strategies to target DTP vulnerabilities, including synthetic lethality approaches, adaptive dosing regimens informed by mathematical modeling, and immune-mediated eradication. Additionally, we highlight synergistic interventions such as lifestyle modifications (e.g., exercise, stress reduction) to suppress pro-tumorigenic inflammation. By integrating mechanistic insights with translational perspectives, this work bridges the gap between DTP biology and clinical strategies, aiming for optimal efficacy and preventing relapse.

In recent years, metabolic reprogramming has emerged as a significant breakthrough in elucidating the onset and progression of gastrointestinal (GI) malignancies. As central regulatory hubs for lipid metabolism, sterol regulatory element binding proteins (SREBPs) integrate dietary metabolic signals and carcinogenic stimuli through subtype-specific mechanisms, thereby promoting malignant tumour phenotypes. In this review, we first present the molecular background, structural characteristics, and posttranscriptional regulatory networks associated with SREBPs. We subsequently describe a systematic analysis of the distinct activation patterns of SREBPs in liver, gastric, colorectal, and other gastrointestinal cancers. Furthermore, we explore targeted intervention strategies for different SREBP subtypes, including small molecule inhibitors (such as fatostatin, which inhibits SREBP cleavage), natural compounds (such as berberine, which modulates the AMPK/mTOR pathway), and statin-mediated inhibition of the mevalonic acid pathway. These strategies may enhance tumour cell sensitivity to chemotherapeutic agents (such as 5-FU, gezil, and tabine) and improve the response to synergistic chemoradiotherapy by reversing adaptive metabolic resistance driven by the tumour microenvironment. Through this review, we hope to provide new insights into precise interventions targeting various subtypes of the SREBP molecule.

Myocardial infarction (MI) is currently one of the most fatal cardiovascular diseases worldwide. The screening, treatment, and prognosis of MI are top priorities for cardiovascular centers globally due to its characteristic occult onset, high lethality, and poor prognosis. MI is caused by coronary artery occlusion induced by coronary atherosclerotic plaque blockage or other factors, leading to ischemic necrosis and apoptosis of cardiomyocytes. Although significant advancements have been made in the study of cardiomyocytes at the cellular and molecular levels, RNA-binding proteins (RBPs) have not been extensively explored in the context of MI. RBPs, as key regulators coordinating cell differentiation and tissue homeostasis, exhibit specific functions in gene transcription, RNA modification and processing, and post-transcriptional gene expression. By binding to their target RNA, RBPs coordinate various RNA dynamics, including cellular metabolism, subcellular localization, and translation efficiency, thereby controlling the expression of encoded proteins. Classical RBPs, including HuR, hnRNPs, and RBM family molecules, have been identified as critical regulators in myocardial hypoxia, oxidative stress, pro-inflammatory responses, and fibrotic repair. These RBPs exert their effects by modulating key pathophysiological pathways in MI, thereby influencing specific cardiac outcomes. Additionally, specific RBPs, such as QKI and fused in sarcoma (FUS), are implicated in the apoptotic pathways activated during MI. This apoptotic pathway represents a significant molecular phenotype in MI, offering novel perspectives and insights for mitigating cardiomyocyte apoptosis and attenuating the progression of MI. Therefore, this review systematically summarizes the role of RBPs in the main pathophysiological stages of MI and explores their potential therapeutic prospects.

<h4>Purpose</h4>Due to the efficient bone targeting of mesoporous silica nanoparticles (MSNs) and polyaspartic acid (PASP), ^99mTc- labeled polyaspartic acid coated mesoporous silica nanoparticles (PASP-mSiO₂-DTPA-^99mTc) are proposed as a potential probe for bone imaging.<h4>Methods</h4>Polyaspartic acid-conjugated silica nanoparticles (PASP-mSiO₂) were synthesized using aqueous carbodiimide chemistry and characterized by ATR-FTR, FE-SEM, EDX, TEM, TGA and XRD. Radiolabeling of the produced nanoassembly with ^99mTc was carried out via a simple DTPA chelation procedure. Aqueous dispersion of the radiolabeled nanoparticles was intravenously injected into normal mice and the bone targeting efficiency was evaluated.<h4>Results</h4>The PASP-mSiO₂ nanoassembly was efficiently synthesized and radiolabeled with ^99mTc with a high radiochemical yield (92 ± 0.5%) and sufficient in vitro stability in PBS and FBS for up to 24 h. In vivo biodistribution studies revealed a significant enhancement of radioactivity bone uptake after intravenous injection of PASP-mSiO₂-DTPA-^99mTc compared to radiolabeled uncoated MSNs (mSiO₂-DTPA-^99mTc), (13 ± 0.6% IA/gram and 5.4 ± 0.4, respectively).<h4>Conclusion</h4>PASP endowed MSNs with enhanced biocompatibility and highly selective bone targeting. Therefore, the proposed PASP-mSiO₂-DTPA-^99mTc nanoassembly has immense potential in the field of bone- imaging via single photon emitting computed tomography (SPECT).

<b>Background:</b> Ferroptosis, a regulated form of cell death characterized by iron-dependent lipid peroxidation, has been implicated in the pathogenesis of asthma. The ferroptosis markers PEBP1 and 15-LO-1 are increasingly recognized as potential biomarkers for asthma. This study investigates the association of these markers with asthma and its severity to evaluate their diagnostic potential. <b>Methods:</b> This cross-sectional study included 45 asthmatic patients and 45 healthy controls. Serum phosphatidylethanolamine-binding protein 1 (PEBP1) and 15-lipoxygenase-1 (15-LO-1) levels were measured using ELISA. Spirometric parameters (FEV₁, FEV₁/FVC ratio, and PEFR) were recorded. A multivariate regression assessed associations between ferroptosis markers and asthma severity. A generalized linear model (GLM) analyzed the relationship between biomarkers (PEBP1 and 15-LO-1) and lung function parameters. A receiver operating characteristic (ROC) analysis evaluated the discriminative capacity of PEBP1 and 15-LO-1. <b>Results:</b> PEBP1 and 15-LO-1 levels were significantly associated with asthma. The multivariate analysis revealed that low PEBP1 levels were strongly associated with asthma and severe asthma (<i>p</i> < 0.001). While elevated 15-LO-1 levels were associated with asthma (<i>p</i> < 0.001), they did not correlate with severity. The ROC analysis demonstrated excellent discriminative capacity for PEBP1 (AUC 0.962, cutoff 1509.8 pg/mL) and 15-LO-1 (AUC 0.895, cutoff 144.8 pg/mL). Lower PEBP1 and higher 15-LO-1 levels were associated with reduced lung function, and lower FEV₁, FEV₁/FVC, and PEF. Older age and female gender were associated with severe asthma. <b>Conclusions:</b> PEBP1 and 15-LO-1 are promising biomarkers for asthma, with PEBP1 showing strong correlations with asthma severity. These findings highlight the potential role of ferroptosis markers in asthma and underscore the need for further longitudinal studies to explore these markers' clinical utility in personalized asthma management.

Recently, there has been a gradual increase in the demand for chicken and eggs. The gut, as the vital place of nutrient digestion and absorption, is highly associated with the development of livestock and poultry and the quality of meat, eggs, and milk. Intestinal stem cells, as an important source of intestinal cell proliferation and renewal, exert a vital effect on repairing injured intestinal epithelial cells and keeping homeostasis. Intestinal stem cell-regulated intestinal epithelial balance is closely controlled and modulated by interlinked developmental loops that maintain cell proliferation and differentiation processes in balance. Some conservative signaling pathways, including the Wnt, Notch, hedgehog, and bone morphogenetic protein (BMP) loops, have been proved to modulate intestinal health in poultry. Meanwhile, studies have revealed the importance of the Hippo pathway in gastrointestinal tract physiology by regulating intestinal stem cells. Moreover, crosstalk between Hippo and other signaling pathways provides tight, yet versatile, regulation of tissue homeostasis. In this review, we summarize studies on the role of the Hippo pathway in the intestine in these physiological processes and the underlying mechanisms responsible via interacting with these signaling pathways and discuss future research directions and potential therapeutic strategies targeting Hippo signaling in intestinal disease. A comprehensive understanding of how these signaling pathways regulate stem cell proliferation, differentiation, and self-renewal will help to understand the regulation of intestinal homeostasis. In addition, it has the capacity for creative ways to govern intestinal damage, enteritis, and associated disorders induced by different factors.

<b>Objective:</b> Epilepsy, a common neurological disorder marked by recurrent seizures often starting in childhood, has a complex etiology. Advances in high-throughput sequencing now confirm that 70-80% of cases have a genetic basis. Accordingly, this study aims to evaluate the clinical relevance of genetic variations detected through epilepsy panels and whole exome sequencing (WES) in pediatric-onset epilepsy patients. <b>Methods:</b> For this study, we enrolled a cohort of pediatric patients involving 205 subjects with a preliminary diagnosis of epilepsy. Targeted next-generation sequencing panels for epilepsy and whole exome sequencing was performed using the NextSeq 500 platform. The results were analyzed with the QIAGEN Clinical Insight bioinformatic platform and were further confirmed and approved by the Human Genome Mutation Database and ClinVar databases. <b>Results:</b> In this study, an epilepsy panel was conducted in 138 patients, and whole exome sequencing was performed in 67 patients. No clinically relevant variants were identified in 29 (21.0%) patients who underwent the epilepsy panel and 27 (40.3%) patients who underwent WES. Variants were detected in 128 different genes in the epilepsy panel group and in 54 different genes in the WES group, with the frequency of these variants limited to one or two patients. <b>Significance:</b> In both the epilepsy panel and WES groups, variants in sodium channel proteins, specifically in the SCN1A, SCN8A, and SCN9A genes, were found to have a high frequency. Collectively, these findings suggest that sodium channel proteins may play an important role in epilepsy.

<h4>Background/objectives</h4>Whole-genome sequencing (WGS) data provide valuable information about the genetic architecture of local livestock but have not yet been applied to Russian native goats, in particular, the Orenburg and Karachay breeds. A preliminary search for selection signatures based on single nucleotide polymorphism (SNP) genotype data in these breeds was not informative. Therefore, in this study, we aimed to address runs of homozygosity (ROHs) patterns and find the respective signatures of selection overlapping candidate genes in Orenburg and Karachay goats using the WGS approach.<h4>Methods</h4>Paired-end libraries (150 bp reads) were constructed for each animal. Next-generation sequencing was performed using a NovaSeq 6000 sequencer (Illumina, Inc., San Diego, CA, USA), with ~20X genome coverage. ROHs were identified in sliding windows, and ROH segments shared by at least 50% of the samples were considered as ROH islands.<h4>Results</h4>ROH islands were identified on chromosomes CHI3, CHI5, CHI7, CHI12, CHI13, and CHI15 in Karachay goats; and CHI3, CHI11, CHI12, CHI15, and CHI16 in Orenburg goats. Shared ROH islands were found on CHI12 (containing the <i>PARP4</i> and <i>MPHOSPH8</i> candidate genes) and on CHI15 (harboring <i>STIM1</i> and <i>RRM1</i>). The Karachay breed had greater ROH length and higher ROH number compared to the Orenburg breed (134.13 Mb and 695 vs. 78.43 Mb and 438, respectively). The genomic inbreeding coefficient (<i>F</i>_ROH) varied from 0.032 in the Orenburg breed to 0.054 in the Karachay breed. Candidate genes associated with reproduction, milk production, immunity-related traits, embryogenesis, growth, and development were identified in ROH islands in the studied breeds.<h4>Conclusions</h4>Here, we present the first attempt of elucidating the ROH landscape and signatures of selection in Russian local goat breeds using WGS analysis. Our findings will pave the way for further insights into the genetic mechanisms underlying adaption and economically important traits in native goats.

Small interfering RNA (siRNA) therapeutics are a new class of drugs that is rapidly expanding to tackle various diseases. Extrahepatic delivery of siRNAs, especially to the parenchyma of solid tumors, is challenging with multiple strategies being explored such as lipid nanoparticle based delivery and ligand conjugation strategies. Here, we report that an albumin-binding dendritic siRNA (D-siRNA) boosts blood circulation time following systemic administration, leading to improved delivery and silencing activity in a melanoma tumor model, in comparison to non-albumin binding lipophilic siRNAs. D-siRNAs increased the tumor-to-liver delivery ratio, including both immune and non-immune cell types within the tumor parenchyma. Using D-siRNAs to target JAK1 expression as an adjuvant to immune checkpoint inhibitors, we found that D-siRNAs was able to enhance PD1 antibody treatment and slow tumor progression of melanoma. Thus, this work demonstrates the utility of D-siRNAs as a systemically administered tumor delivery strategy, enabling the use of siRNAs as chemotherapeutic agents. Further mechanistic studies into the role of JAK1 in melanoma pathology and progression may expand this into additional targets as potential treatments.

<h4>SUMMARY</h4> This study explores the inflammatory response observed in pancreata and pancreatic lymph node (pLN) samples obtained throughout the natural history of type 1 diabetes (T1D) including non-diabetic individuals and non-diabetic autoantibody positive individuals with high susceptibility using spatial transcriptomics (ST). Integration of ST with public single-cell RNA sequencing data enabled interrogation of transcriptional alterations in T1D pathogenesis across both tissues and cellular scales. In the T1D pancreas, we observed global upregulation of multiple inflammation-associated transcripts, including regenerating islet-derived ( REG ) family genes, complement factor 3 ( C3 ), SOD2 , and OLFM4 , and highlighted cellular candidates potentially contributing to these signatures. Within the T1D pLN, we observed spatially restricted upregulation of lymphotoxin-β ( LTB ) alongside follicular dendritic cell (FDC)-associated transcripts including FDCSP , CLU , and FCER2 . Collectively, these findings highlight distinct inflammation signatures in the pancreas and regional pLN which can help inform the development of future therapeutic interventions.

High-throughput massively parallel reporter assays (MPRAs) and phenotype-rich in vivo transgenic mouse assays are two potentially complementary ways to study the impact of noncoding variants associated with psychiatric diseases. Here, we investigate the utility of combining these assays. Specifically, we carry out an MPRA in induced human neurons on over 50,000 sequences derived from fetal neuronal ATAC-seq datasets and enhancers validated in mouse assays. We also test the impact of over 20,000 variants, including synthetic mutations and 167 common variants associated with psychiatric disorders. We find a strong and specific correlation between MPRA and mouse neuronal enhancer activity. Four out of five tested variants with significant MPRA effects affected neuronal enhancer activity in mouse embryos. Mouse assays also reveal pleiotropic variant effects that could not be observed in MPRA. Our work provides a catalog of functional neuronal enhancers and variant effects and highlights the effectiveness of combining MPRAs and mouse transgenic assays.

<h4>Background</h4>Head and neck squamous cell carcinomas (HNSCs) are among the most common tumors worldwide. Despite the availability of various diagnostic and therapeutic strategies, the incidence and mortality rates of HNSC remain high. Cancer-associated fibroblasts (CAFs), as a major component of the tumor microenvironment, exhibit diverse biological characteristics in terms of origin, genetics, and phenotype, and have been increasingly recognized for their roles in tumor progression.<h4>Methods</h4>To investigate the potential role of CAFs in HNSC, we performed a comprehensive bioinformatics analysis based on the TCGA HNSC cohort. We applied single-sample gene set enrichment analysis (ssGSEA), single-cell RNA sequencing (scRNA-seq) analysis, differential expression analysis, Cox regression, LASSO regression, and pathway enrichment analysis to identify CAF-related genes and assess their prognostic value.<h4>Results</h4>We successfully identified a set of CAF-related genes and stratified the HNSC patients into high- and low-CAF groups. Based on the expression of these genes, we constructed a prognostic prediction model using LASSO and multivariate Cox regression analyses. A nomogram integrating the risk score and clinical characteristics was developed to improve individualized survival prediction. Enrichment analysis revealed that the type I interferon signaling pathway, cellular response to type I interferon, defense response to symbiont, and extracellular matrix organization were significantly associated with CAFs in HNSC.<h4>Conclusion</h4>Our study provides a novel CAF-based prognostic model and nomogram for predicting patient outcomes in HNSC. These findings highlight the importance of CAFs in the tumor microenvironment and their potential as therapeutic and prognostic biomarkers.

<h4>Background</h4>How parental alcohol use disorder and liver disease-related mortality influence the risk and the outcomes of alcohol-associated hepatitis (AH) in the offspring is unknown.<h4>Methods</h4>We analyzed data from 2 prospective observational studies of AH cases and heavy drinking controls (HDCs). Family history of parental alcohol use disorder and liver disease mortality was assessed at the study entry. Logistic regression and Cox proportional hazard models were used to assess the influences of family history on AH development and outcome.<h4>Results</h4>Data from 1356 participants in two prospective cohorts (926 AH cases and 430 HDC) were combined and analyzed. Parental alcohol use disorder was found in 56.9% of AH cases and 61.1% of HDC; parental death due to liver disease was reported in 7.5% of AH cases and 5.7% of HDC. Multivariable logistic regression showed that parental liver disease-related mortality was associated with more than a doubled risk of AH development in the offspring after controlling for their demographic characteristics and drinking behavior (OR=2.26, 95% CI: [1.22, 4.20]). Moreover, among the AH cases, having a parent die of liver disease significantly increased the 90-day mortality of study participants after adjusting for the effects of other risk factors (HR=2.26, 95% CI: [1.05, 4.86]).<h4>Conclusions</h4>The study highlights the influences of parental death due to liver disease on AH development and mortality. Identifying patients at risk of AH through family history might help facilitate discussions on reducing alcohol consumption.

Action potentials are initiated and modulated at the axon initial segment (AIS) by highly clustered ion channels. Voltage-gated Kv1 potassium channels underlie most outward AIS K⁺ current. AIS Kv1 channels exist in a large protein complex including ADAM22, Caspr2, and LGI1. However, their clustering mechanisms remain unknown. Because Kv1 channels have a highly conserved PDZ-binding motif, we used CRISPR-based genome editing to screen 18 PDZ domain-containing proteins identified in our previous AIS proximity proteome for their AIS localization. Among these, we found that the scaffolding proteins SCRIB and PSD93 are highly enriched at the AIS. Using CRISPR-mediated knockout, cell surface clustering assays, and coimmunoprecipitation, we show that SCRIB and PSD93 bind to and are required for AIS Kv1 channel clustering, whereas SCRIB links the AIS Kv1 channel protein complex to the master AIS scaffolding protein AnkyrinG. These results define a hierarchy of scaffolding proteins that combine to cluster AIS Kv1 channels.

The self-renewal of intestinal stem cells (ISCs) is essential for maintaining intestinal homeostasis and ensuring regeneration of the intestinal epithelium. However, whether small nucleolar RNAs participate in the regulation of ISC self-renewal remains unclear. Here, we identified a small nucleolar RNA (<i>Snora54</i>) that was highly expressed in the nucleolus of ISCs. <i>Snora54</i> knockout enhanced the self-renewal capacity of ISCs and intestinal regeneration. Mechanistically, in a steady state, highly expressed <i>Snora54</i> anchored the nucleolar protein Lyar in the nucleolus of ISCs, preventing Lyar from translocation into the nucleoplasm. Thereby, Lyar failed to recruit on the <i>Notch2</i> promoter region in the nucleoplasm to promote <i>Notch2</i> transcription, leading to suppression of ISC self-renewal. By contrast, with deletion of <i>Snora54</i>, Lyar translocated to the nucleoplasm of ISCs where it enriched on the <i>Notch2</i> promoter to initiate its transcription resulting in the activation of Notch2 signaling pathway. Therefore, <i>Snora54</i> negatively regulates self-renewal of ISCs and gut regeneration via suppression of Notch2 signaling.

Type 2 inflammation is an essential defense mechanism of the innate and adaptive immune systems, but when dysregulated, it can cause chronic atopic diseases like allergic asthma and atopic dermatitis. Thymic stromal lymphopoietin (TSLP) helps drive type 2 inflammation by guiding T cells toward a type 2 helper cell (T_H2) subtype and stimulating B cells' antibody production. Fibronectin (FN) has recently been found at elevated levels in the plasma of children with atopic dermatitis and shown a potential proinflammatory role in bronchial epithelium tissue models. Both proteins' surface charges suggest potential interaction with charged molecules. Seeking new strategies against type 2 inflammation, we found that negatively charged polyglycerol sulfates strongly bind to TSLP and FN. We confirmed that these molecules inhibit inflammation by reducing the TSLP-mediated type 2 polarization of CD4⁺ T cells. We found that adding polyglycerol sulfate to FN-triggered inflamed bronchial epithelium models reduced TSLP expression and interleukin 6 secretion.

Despite the STING-type-I interferon pathway playing a key role in effective anti-tumor immunity, the therapeutic benefit of direct STING agonists appears limited. In this study, we use several artificial intelligence techniques and patient-based multi-omics data to show that Ectonucleotide Pyrophosphatase/Phosphodiesterase 1 (ENPP1), which hydrolyzes STING-activating cyclic GMP-AMP (cGAMP), is a safer and more effective STING-modulating target than direct STING agonism in multiple solid tumors. We then leverage our generative chemistry artificial intelligence-based drug design platform to facilitate the design of ISM5939, an orally bioavailable ENPP1-selective inhibitor capable of stabilizing extracellular cGAMP and activating bystander antigen-presenting cells without inducing either toxic inflammatory cytokine release or tumor-infiltrating T-cell death. In murine syngeneic models across cancer types, ISM5939 synergizes with targeting the PD-1/PD-L1 axis and chemotherapy in suppressing tumor growth with good tolerance. Our findings provide evidence supporting ENPP1 as an innate immune checkpoint across solid tumors and reports an AI design-aided ENPP1 inhibitor, ISM5939, as a cutting-edge STING modulator for cancer therapy, paving a path for immunotherapy advancements.

The tumorigenesis of small intestinal neuroendocrine tumors (siNETs) is not understood and comprehensive genomic and transcriptomic data sets are limited. Therefore, we performed whole genome and transcriptome analysis of 39 well differentiated siNET samples. Our genomic data revealed a lack of recurrent driver mutations and demonstrated that multifocal siNETs from individual patients can arise genetically independently. We detected germline mutations in Fanconi anemia DNA repair pathway (FANC) genes, involved in homologous recombination (HR) DNA repair, in 9% of patients and found mutational signatures of defective HR DNA repair in late-stage tumor evolution. Furthermore, transcriptomic analysis revealed low expression of the transcriptional repressor REST. Summarizing, we identify a novel common transcriptomic signature of siNETs and demonstrate that genomic alterations alone do not explain initial tumor formation, while impaired DNA repair likely contributes to tumor evolution and represents a potential pharmaceutical target in a subset of patients.