The role of hepatic insulin resistance (HIR) in the development of fatty liver, diabetes and cardiovascular diseases is well known, yet the molecular basis of HIR remains unclear, limiting targeted therapeutic strategies. Here we show that insulin signalling-inactivated phosphorylated GSK-3β (p-GSK-3β) is revitalized via reactive oxygen species-mediated sulfenylation, leading to glycogenesis termination and gluconeogenesis initiation, two hallmarks of HIR. Mechanistically, sulfenylated or 'oxidatively activated' p-GSK-3β regains the enzymatic activity to phosphorylate liver glycogen synthase, thereby blocking glucose storage. This activated p-GSK-3β can further phosphorylate insulin-suppressed Forkhead box O1, thus liberating its transcriptional activity to promote the expression of gluconeogenic enzymes. Notably, this dual-pathway mechanism is conserved in clinically relevant human liver samples and organoids. These findings elucidate the molecular mechanism by which HIR is formed and provide potential strategies against HIR by targeting sulfenylated or 'oxidatively activated' p-GSK-3β.

Vascular remodeling is a characteristic pathological feature of various vascular diseases, including atherosclerosis, restenosis following vascular injury, hypertension, and aneurysms. The phenotypic switching of vascular smooth muscle cells (VSMCs) acts as a key driver of vascular remodeling. Under specific pathological stimuli, VSMCs rapidly transition from a contractile to a dedifferentiated phenotype, characterized by enhanced proliferation, migration, and secretory activity. Chromatin remodeling, a core mechanism of epigenetic regulation, orchestrates dynamic changes in chromatin structure and function through ATP-dependent remodeling complexes, histone-modifying enzymes, and DNA methyltransferases. These components collectively translate mechanical stress, metabolic disturbances, and inflammatory signals into reversible epigenetic modifications, thereby precisely regulating VSMC phenotypic switching. As such, chromatin remodeling represents a critical node for therapeutic intervention in vascular remodeling-related diseases. In recent years, a growing body of research has focused on the role of chromatin remodelers in regulating VSMC phenotype. In this review, we focus on the roles of ATP-dependent chromatin-remodeling factors and chromatin-modifying enzymes in the control of gene expression of VSMC phenotype switching. Firstly, we summarize the latest insights into chromatin remodeling and VSMC phenotypic switching, and then discuss recent advances in the identification and functional characterization of chromatin remodeling molecules, emphasizing their implications for VSMC behavior. Finally, we highlight the translational potential of targeting chromatin remodelers in the development of clinical therapies for vascular remodeling diseases and outline future directions for research in this field.

Optimizing oocyte maturation in high ovarian responders undergoing in vitro fertilization is crucial for reproductive outcomes and minimizing ovarian hyperstimulation syndrome (OHSS). It remains uncertain whether a gonadotropin-releasing hormone agonist (GnRH-a) combined with low-dose hCG (dual trigger) provides advantages over GnRH-a. This meta-analysis evaluated the efficacy and safety of dual trigger versus GnRH-a trigger in high ovarian responders. A systematic search of PubMed, Embase, Web of Science, the Cochrane Library, and China National Knowledge Infrastructure was conducted to identify randomized controlled trials (RCTs) and cohort studies comparing dual trigger with GnRH-a trigger alone in high responders. The primary outcomes were metaphase II (MII) oocyte rate and moderate or severe OHSS. Secondary outcomes included the number of retrieved oocytes, embryos, and high-quality embryos; normal fertilization rate; clinical pregnancy rate (CPR); miscarriage rate; and live birth rate. The study analyzed binary and continuous variables using relative risk (RR) and weighted mean difference (WMD), with 95

Background: Recurrent implantation failure (RIF) remains a major challenge in in vitro fertilization-embryo transfer (IVF-ET), particularly in patients with unexplained infertility. Although immune dysregulation is increasingly recognized as a contributor to female infertility, evidence-based immunomodulatory strategies remain limited. This study evaluated the efficacy and safety of cyclosporine A (CsA) in RIF patients with immunological abnormalities. Methods: In a retrospective analysis, 223 women with repeated IVF-ET failures and positive autoantibody markers were included. Among them, 134 patients underwent 252 IVF-ET cycles receiving either conventional therapy (aspirin and/or corticosteroids) or conventional therapy combined with CsA (25-50 mg twice daily). Subsequently, a prospective cohort of 55 patients undergoing 60 embryo transfers received CsA treatment initiated 3 months before embryo transfer and continued until 12 weeks of gestation. The primary outcomes were implantation success and live birth rates, with maternal and neonatal safety assessed. Findings: In the retrospective cohort, CsA supplementation significantly improved IVF-ET outcomes, with a success rate of 66.7% compared with 0.5% in the conventional treatment group (P<0.001). These findings were confirmed in the prospective cohort, where 47 of 60 embryo transfers resulted in live births (78.3%). CsA treatment was associated with immunological improvements, including correction of CD4+T cell deficiency and normalization of natural killer cell proportions. No serious maternal adverse events, including hepatic or renal dysfunction, were observed during follow-up. Interpretation: CsA therapy improved reproductive outcomes in RIF patients with immune abnormalities while demonstrating favorable maternal-fetal safety. These findings support CsA as a potential immunomodulatory treatment strategy for RIF.

Gut microbiome plays a pivotal role in modulating immunotherapy responses in colorectal cancer (CRC) treatment. While individual enterobacteria have been identified as enhancers of anti-PD-1/anti-PD-L1 therapy, the synergistic effects of multiple probiotic strains remain insufficiently explored. In this study, we investigated the therapeutic potential of Tumor-Suppressing Multi-Enterobacteria (TSME), a consortium of nine beneficial intestinal probiotic strains, in enhancing anti-PD-1/anti-PD-L1 therapy for microsatellite stable (MSS) CRC. Using a tumor-bearing mouse and employing techniques including flow cytometry, immunohistochemistry, ELISA, and genomic sequencing, we found that TSME significantly improved the efficacy of immune checkpoint inhibitors (ICIs) by optimizing tumor immune and microbe microenvironment. Specifically, the addition of TSME increased CD8+ T cell infiltration and reshaped cytokine profiles, including reducing pro-inflammatory cytokines (IL-17, IL-1β, IL-6, and TNF-α) while elevating anti-inflammatory factors (IFN-γ). Moreover, TSME significantly up-regulated key immune pathways, including TNF signaling, cytokine-cytokine receptor interaction, and JAK-STAT signaling. In addition, TSME restructured the gut microbiome, increasing the abundance of beneficial bacteria such as Akkermansia and Alistipes. These findings highlight the synergistic effect of the multi-strain probiotics in enhancing ICI efficacy. Well-formulated probiotic consortia offer a promising strategy for enhancing immunotherapy outcomes in MSS CRC and advancing broader implementation of microbiome-assisted precision oncology.

Despite the pleiotropic capacities of cytokines in modulating cell behaviors, their therapeutic application in cancer remains challenging. Here, we show that the IFN-γ/IFN-β/TGF-β cocktail integrates these three signals with a cytosolic pore-forming protein, gasdermin E (GSDME), and synergistically drives its delivery into the lysosomes of pancreatic adenocarcinoma (PDAC) tumor-repopulating cells (TRCs), where GSDME is cleaved to mediate lysosomal pore formation. Mechanistically, IFN-γ signaling phosphorylates GSDME, enabling phosphorylated GSDME (p-GSDME) to bind the Golgi transmembrane protein TMED10 and subsequently traffic to lysosomes, where cathepsin D cleaves it into active N-GSDME, which induces lysosomal decomposition in TRCs. In parallel, IFN-β activates STAT1/STAT3 to upregulate cathepsin D expression, whereas TGF-β enhances GSDME phosphorylation by downregulating PPP1R3G, a regulatory subunit of protein phosphatase 1. Using lipid-hybrid nanoparticle-delivered mRNA technology, the tri-cytokine cocktail demonstrated therapeutic efficacy against orthotopic PDAC in mice and PDX models, highlighting its translational potential for PDAC patients.

OBJECTIVE:To investigate the effects of irisin on the Wnt/β-catenin signaling pathway in the endometrium of obese rats. METHODS:Seventy-two female Sprague Dawley rats of 5 weeks old were randomly divided into the normal group (n = 24) and obese group (n = 48). The obese group was fed a high-fat diet, while the normal group was fed a conventional diet. After successful modeling, obese rats were randomly divided into the model group (n = 24) and irisin group (n = 24). The irisin group was given an intraperitoneal injection of recombinant irisin every day, while the model group and the normal group were intraperitoneally injected with an equal volume of normal saline. Then, female rats in estrus were mated with male rats. Twelve rats were sacrificed in each group on Day 5 and day 10 of pregnancy. The pregnancy rate, average number of blastocysts, histological and morphological changes were observed; the expression of Leukemia Inhibitory Factor (LIF), integrin αvβ3, Wnt4, and β-catenin in the uterus of rats was detected. RESULTS:Compared with the normal group, levels of pregnancy rate, average number of blastocyst, LIF, integrin αvβ3, Wnt4, and β-catenin in the endometrium of the model group rats were significantly decreased. After irisin intervention, compared with the model group, levels of pregnancy rate, average number of blastocyst, LIF, integrin αvβ3, Wnt4, and β-catenin in the endometrium of the irisin group were significantly increased. CONCLUSION:Irisin may improve endometrial receptivity in obese rats, potentially involving the regulation of the Wnt/β-catenin signaling pathway.

Reactive oxygen species (ROS) dynamics exhibits rhythmic oscillations in cancer cells but how this rhythm influences tumorigenesis and therapeutic responses remains unclear. Here we found coexistence of ROS rhythmicity and rhythm loss in tumor samples. Under low-ROS conditions, indoleamine 2,3-dioxygenase 1 (IDO1), an immune-checkpoint molecule, binds to KEAP1 for proteasomal degradation in the nucleus. In contrast, elevated ROS levels drive IDO1 translocation into the cytosol, where it binds mitochondria-released heme to form an active holoenzyme. This holoenzyme catalyzes tryptophan to kynurenine that allosterically activates glucose-6-phosphate dehydrogenase, enhancing NADPH production and promoting ROS clearance. However, in hypoxic tumor microenvironments, ROS rhythmicity is lost. Compensating for this, hypoxic tumor cells mobilize the sulfenylated aryl hydrocarbon receptor (AhR)-mediated glycogenolysis pathway to manage disordered ROS accumulation, maintaining elevated ROS levels that favor tumor growth. Dual inhibition of IDO1 and AhR significantly prolongs survival of NSG mice, highlighting enforced disruption of ROS rhythm as a common therapeutic strategy.

At present, the known genetic causes of abnormal oocyte development can only account for a minority of female infertility. In our previous study, the mutation c.1101 C > G, p.Tyr367* in discs large-associated protein 5 (DLGAP5) was identified as a novel genetic cause of human oocyte maturation abnormality and female infertility. The present study aimed to validate the function of DLGAP5 in oocyte maturation and further explore the underlying mechanism by which DLGAP5 regulates oocyte meiosis. Cell experiments elucidated that DLGAP5 participates in cell division, and its depletion induced G2/M arrest. The depletion of DLGAP5 in human oocytes by microinjection of siRNAs resulted in abnormal spindle morphology and oocyte maturation defects, exhibiting reduced germinal vesicle breakdown and polar body 1 extrusion rate. In addition, a similar phenotype of abnormal oocyte development was observed in Dlgap5-deficient mouse oocytes, which could be rescued by DLGAP5 cRNA microinjection. Furthermore, single-cell RNA Sequencing showed Dlgap5 knockout altered expression of genes involving in meiosis process in oocytes and deactivated PI3K-AKT signaling pathway. And PI3K-AKT activators facilitated the oocyte maturation resumption in Dlgap5-deficient mice. DLGAP5 has been demonstrated to regulate the process of oocyte maturation via the activation of PI3K-AKT pathway. It reinforces the significant role of DLGAP5 function in oocyte maturation regulation and reveals the underlying mechanism. It provides crucial insights into clinical consultation, genetic diagnosis, and treatment strategies among infertile patients.

Recurrent implantation failure (RIF) remains a significant clinical challenge in assisted reproductive technology. Its etiology is often attributed to inadequate endometrial receptivity, yet the underlying molecular mechanisms are poorly characterized, leading to a lack of reliable diagnostic biomarkers and targeted treatments. We conducted RNA sequencing on endometrial biopsies obtained during the window of implantation from 9 RIF patients and 13 fertile controls. Differential gene expression analysis was performed, followed by comprehensive functional enrichment analysis (GO (Gene Ontology), KEGG (Kyoto Encyclopedia of Genes and Genomes), and GSEA (Gene Set Enrichment Analysis)). The expression of key candidate genes was validated using qRT-PCR, Western blotting, and immunohistochemistry. RNA sequencing revealed 103 differentially expressed genes in RIF endometrium. GO and KEGG analyses consistently highlighted significant enrichment in immune-related processes and pathways, particularly acute inflammatory response, leukocyte-mediated immunity, and complement activation. GSEA highlighted strong enrichment of immune pathways including natural killer cell-mediated cytotoxicity and interleukin signaling, which emphasized dysregulation of immune and inflammatory processes. Key upregulated molecules including EDNRB, AACT, REL, USP18, and ANG were validated experimentally, suggesting their potential as biomarkers for RIF. Our study delineates a distinct immune-dysregulated transcriptomic profile in the endometrium of RIF patients, providing crucial insights into the pathogenesis of this condition. The validated genes, EDNRB, AACT, REL, USP18, and ANG, represent promising biomarkers with significant potential for improving the diagnosis and future therapeutic strategies for RIF.

Sonic hedgehog subgroup medulloblastoma (SHH-MB), an aggressive pediatric brain tumor that originates from granule neuron precursors, faces the challenge of poor treatment owing to its unclear molecular mechanisms. Here, we show that sialic acid-binding immunoglobulin-like receptor 15 (Siglec-15), an immunosuppressive membrane protein, is upregulated and mediates SHH-MB growth through its translocation to the lysosomal membrane. We found that SHH-MB cells use the cation-independent mannose 6-phosphate receptor (CI-MPR) to transport Siglec-15 from the trans-Golgi network (TGN) to lysosomes, where Siglec-15 induces lysosomal Ca2+ release by interacting with mucolipin TRP cation channel 1 (TRPML1), leading to the nuclear translocation of the transcription factor EB (TFEB). Blockade of Siglec-15, TRPML1 or TFEB hinders SHH-MB growth in vitro and in vivo. Importantly, aryl hydrocarbon receptor (AhR), a cytoplasmic transcription factor, upregulates Siglec-15 expression. AhR inhibition by CH-223191 or StemRegenin 1 (SR1) achieved therapeutic efficacy against orthotopic SHH-MB xenografts in mice. These findings reveal an essential role for the AhR-siglec-15 axis in SHH-MB development, providing a potential strategy for SHH-MB treatment.

Coronary artery disease (CAD), one of the most prevalent cardiovascular diseases, is a critical health issue that affects millions of individuals worldwide. It has been reported that miR-146b-5p exhibited a strong correlation with inflammatory responses and atherosclerosis. However, its association with the incidence and severity of CAD has not been substantiated in a large cohort. In the study, we focus on the expression of miR-146b-5p in peripheral blood mononuclear cells (PBMCs) of patients with CAD and preliminarily investigate its function and underlying mechanism. The study encompassed a total of 452 participants, consisting 295 patients with CAD and 157 individuals without CAD. Quantitative reverse transcription–polymerase chain reaction (qRT–PCR) was performed to assess miR-146b-5p expression in PBMCs. We found that miR-146b-5p was significantly increased in PBMCs of patients with CAD compared with the control group. Binary logistic regression revealed that miR-146b-5p was associated with CAD. Receiver Operation Characteristic (ROC) analysis showed that the sensitivity and specificity of miR-146b-5p in discriminating CAD patients from non-CAD patients were meaningful. Subsequent subgroup analysis showed that miR-146b-5p was related to the severity of CAD. Furthermore, gain- and loss-of-function experiments in THP-1 cells showed that miR-146b-5p inhibited inflammation, cell proliferation, and migration. Mechanically, miR-146b-5p was involved in the classical NF-κB inflammatory pathway by directly targeting IKKβ. Our study revealed that miR-146b-5p was higher in the PBMCs of CAD patients than non-CAD individuals, and established a correlation between miR-146b-5p and occurrence and severity of CAD. In addition, the inflammatory role of miR-146b-5p is mediated by targeting IKKβ.

Downstream of T cell receptor (TCR) signaling, proliferation and differentiation programs are thought to be linked to maintain T cell homeostasis. This biology, however, also leads to the in vitro generation of suboptimal chimeric antigen receptor (CAR) T cells. Here, we show that proliferation and differentiation programs can be decoupled by fibrin matrix-based mechanical signaling, leading to abundant generation of undifferentiated stem cell-like CAR (stem-CAR) T cells. These stem-CAR T cells expressed NANOG, SOX2, and TCF1, exhibited persistent cytolysis in tumor cells in vitro, and achieved optimal efficacy in solid tumor models of breast, pancreatic, and brain cancer in vivo. Mechanistically, the fibrin matrix activated β2 integrin to recruit 14-3-3ζ, leading to Yes-associated protein (YAP) phosphorylation and inactivation. Consequently, YAP inactivation derepressed the transcription factor MafG. MafG then transactivated stemness genes, thereby generating stem-CAR T cells. These findings suggest a mechanical approach to manufacturing stem-CAR T cells, potentially improving CAR T cell therapeutic efficacy for cancer treatment.

OBJECTIVES:This study aimed to evaluate the efficiency of endometrial receptivity testing (ERT) in improving pregnancy outcomes for patients with recurrent implantation failure (RIF), and to investigate the incidence of implantation window displacement. METHODS:Conducted between April 2021 and August 2022, at a university-affiliated reproductive center, the study included 85 RIF patients who had failed to achieve pregnancy after three embryo transfers. As part of a multicenter prospective cohort study (ChiCTR2200059342), 45 patients underwent ERT-guided frozen single blastocyst transfer, while 40 received standard treatment without ERT. Endometrial preparation was performed using a hormone replacement therapy (HRT) protocol in the cycle preceding transfer. Endometrial sampling for ERT, including RNA sequencing, artificial intelligence, and discriminant analysis of endometrial receptivity, was conducted on day 5 after progesterone administration to determine the implantation window. The same HRT protocol was used in the transfer cycle, with embryo transfer timed according to the ERT-calculated window. RESULTS:Our data showed that 28.07% of patients exhibited a displaced implantation window, all characterized by pre-receptive endometrium. The ERT group had significantly higher clinical pregnancy rates (57.78% vs. 35.00%, p = 0.036) and live birth rates (53.33% vs. 30.00%, p = 0.030) compared with the non-ERT group. CONCLUSION:Our findings suggest that approximately one-third of RIF occurrences may be due to endometrial factors, and ERT-guided personalized embryo transfer significantly improves pregnancy outcomes, underscoring its value in reproductive medicine.

Glucose-6-phosphate (G6P) is a key metabolic molecule that regulates reactive oxygen species (ROS) homeostasis by initiating the pentose phosphate pathway (PPP) to generate nicotinamide adenine dinucleotide phosohate (NADPH) that converts hydrogen peroxide (H2O2) to water by providing hydrogen. While both glucose phosphorylation and glycogenolysis result in G6P production, here we show that G6P derived from glycogenolysis, rather than glucose phosphorylation, flows to PPP for ROS clearance in CD8+ memory T (Tm) cells and inflammatory macrophages. Mechanistically, glycogenolysis-produced glucose-1-phosphate (G1P) allosterically induces G6P dehydrogenase (G6PD) binding to glycogen, which together undergo liquid-liquid phase separation (LLPS) and recruit PPP enzymes, resulting in a compartmentalized reaction cascade. Based on mechanistic elucidation, we demonstrated that G1P can act as an antitumor immunotherapeutic agent by modulating memory fitness and maintenance of tumor-reactive CD8+ T cells in mice. These findings revealed an unusual function of glycogen metabolism, which is of paramount importance in the regulation of PPP and redox homeostasis in cells.

To develop and validate a CT radiomics model for predicting microsatellite instability (MSI) status in preoperative gastric cancer (GC) patients and to explore the underlying immune infiltration pattern of the radiomics model. This study used three retrospective datasets from Tongji Hospital (n = 304, training set), Xiangyang Central Hospital (n = 48, external testing set 1) and public datasets from The Cancer Imaging Archive (TCIA) (n = 43, external testing set 2). The preoperative contrast-enhanced CT images of GC were evaluated. Radiomics features were extracted and selected to construct the radiomics model in the training set, and further validated in the other two external testing sets. The outcome cohort, including 68 advanced unresectable GC patients receiving immunotherapy, was used to assess the predictive value of the radiomics model for treatment response and outcomes. We analyzed RNA-sequencing data from TCIA to investigate the underlying genomics characterization and immune infiltration spectrum of the radiomics model. Four radiomic features were ultimately selected to develop the radiomics model. The model demonstrated good predictive performance for MSI status, achieving AUCs of 0.952, 0.835, and 0.879 in the training set and the two external testing sets, respectively. Radiomics scores (Radscores) was an independent predictor for PFS in the outcome cohort (HR: 0.145; 95

The long-term association between total energy intake and clinical outcomes in patients with metabolic dysfunction-associated steatotic liver disease (MASLD) remains unclear. The present study aimed to explore the associations between total energy intake and mortality in MASLD patients and to evaluate whether the associations differ by sex. A total of 2,466 MASLD patients from the Third National Health and Nutrition Examination Survey was included. Total energy intake was assessed using 24-hour dietary recall. Mortality was ascertained by linkage to National Death Index records through 31 December 2019. Multivariable Cox proportional hazards model was used to estimate the association between total energy intake and mortality. In MASLD patients, both low and high total energy intake were significantly associated with elevated risk of all-cause mortality. Compared with moderate total energy intake (2,000-3,000 kcal/day for males and 1,600-2,400 kcal/day for females), the HRs (95% CIs) for low and high total energy intake were 1.27 (1.05-1.53; P = 0.01) and 1.40 (1.03-1.92; P = 0.03), respectively. A significant interaction was demonstrated between sex and total energy intake for all-cause mortality (P value for interaction = 0.03). In males, both low and high total energy intake were significantly associated with elevated risk of all-cause mortality. The HRs (95% CIs) for low and high total energy intake were 1.35 (1.02-1.80; P = 0.04) and 1.54 (1.05-2.28; P = 0.03), respectively. However, no significant association was observed in females. The HRs (95% CIs) for low and high total energy intake were 1.14 (0.86-1.52; P = 0.34) and 1.14 (0.81-1.61; P = 0.46), respectively. These findings provide novel evidence supporting sex-specific dietary guidelines for MASLD, with particular emphasis on maintaining moderate energy intake to mitigate mortality risk in male patients.

Fine particulate matter 2.5 (PM2.5) is a prevalent atmospheric pollutant that is closely associated with asthma. Elderly patients have a high incidence of asthma with a long course of illness. Our previous studies revealed that exposure to PM2.5 diminishes lung function and exacerbates lung damage in elderly rats. In the present study, we investigated whether PM2.5 exposure influences susceptibility to allergic asthma in elderly rats. Brown-Norway elderly rats were treated with ovalbumin (OVA) for different durations before and after PM2.5 exposure. The results from pulmonary function tests and histopathology indicated that early exposure to allergens prior to PM2.5 exposure increased susceptibility to airway hyperresponsiveness and led to severe lung injury in elderly asthmatic rats. Cytokine microarray analysis demonstrated that the majority of cytokines and chemokines were upregulated in OVA-treated rats before and after PM2.5 exposure. Cytological examination showed no change in eosinophil (EOS) counts, yet the amounts of neutrophils (NEU), white blood cells (WBC), lymphocytes (LYM), and monocytes (MON) in the lung lavage fluid of OVA-treated rats were significantly higher than those in control rats before and after PM2.5 exposure, suggesting that PM2.5 affects noneosinophilic asthma in elderly rats. ELISA results from the plasma and lung lavage fluid revealed that the levels of IgG1, IgE, IgG2a and IgG2b were significantly elevated in OVA-treated rats, whereas the level of IgG2b in the lung lavage fluid was significantly lower in rats treated with OVA prior to PM2.5 exposure compared to those treated afterward. A non-targeted metabolomic analysis of plasma identified 202 metabolites, among which 31 metabolites were differentially abundant. Ten metabolites and 11 metabolic pathways were uniquely detected in OVA-treated rats before PM2.5 exposure. Specifically, there were positive or negative correlations between the levels of Th2-associated cytokines (IL-4, IL-5, and IL-13) and six metabolites in the OVA-treated group before PM2.5 exposure, whereas the levels of IL-4 and IL-5 were negatively correlated with five metabolites in the OVA-treated group after PM2.5 exposure. Our findings suggest that PM2.5 exposure could influence the susceptibility of allergic asthma in response to allergens in elderly rats, potentially through changes in plasma metabolites.