This special issue comprises one review and seven original articles covering diverse topics in reproductive medicine,including the microbiota-metabolite axis in endometriosis,BDE47-induced ferroptosis in spermatocytes,human papillomavirus (HPV) and fertility outcomes in endometriosis,metabolomic biomarkers for azoospermia stratification,gene function validation in male reproduction,endometrial preparation protocols and the vaginal microbiota,bisphenol A (BPA)-induced developmental cardiotoxicity,and diagnostic criteria for gestational diabetes.
This study aimed to evaluate the modifying effects and joint associations of handgrip strength (HGS) and bone mineral density (BMD) with all-cause mortality, with an exploratory analysis of cardiovascular disease (CVD) mortality. A primary analysis cohort ( n = 11150) and a replication cohort ( n = 5952) were included. Cox proportional hazards regression, restricted cubic spline analysis, and interaction analysis were used to examine the associations of BMD with mortality across relative HGS groups. Relative HGS was defined as absolute HGS divided by body weight. The joint effects of BMD-HGS combinations were assessed. During a median follow-up of 9.75 and 6.59 years, the primary analysis and replication cohorts recorded 1345 and 352 all-cause deaths, respectively. In the primary analysis cohort, low BMD was associated with an increased all-cause mortality risk in both the low and high relative HGS groups (hazard ratio [HR] = 1.26, 95% confidence interval [CI]: 1.04-1.52 and HR = 1.42, 95% CI: 1.16-1.74, respectively), with consistent findings in the replication cohort. No significant interaction was found between relative HGS and BMD ( P > 0.05). Participants with low BMD and low relative HGS had a 59%-62% higher risk of all-cause mortality than those with high BMD and high relative HGS in both cohorts, and the estimated PAR% for this combined exposure was 10.8%-13.1%. Results for CVD mortality were inconsistent across cohorts. In conclusion, low BMD with low handgrip strength is consistently associated with increased all-cause mortality, suggesting combined screening may benefit high-risk individuals.
Colorectal cancer (CRC) represents a leading global malignancy. This study investigated the role of differentiated embryonic chondrocyte expressed gene 1 (DEC1) in interleukin-6 (IL-6)-induced invasion and migration of CRC. In vitro, IL-6 upregulated DEC1 expression and modulated the levels of epithelial-to-mesenchymal transition (EMT)-associated proteins in CRC cells. DEC1 overexpression amplified the IL-6-induced invasion and migration phenotypes, increasing the expression of N-cadherin and vimentin but reducing E-cadherin expression. Conversely, DEC1 knockdown attenuated these phenotypes, decreasing the expression of N-cadherin and vimentin but increasing E-cadherin expression. These findings suggest that IL-6 induced EMT through increasing DEC1. Moreover, DEC1 overexpression increased the levels of basal and IL-6-stimulated phosphatidylinositol 3 kinase (PI3K) p110α, p-Akt, and p-p65, indicating activation of the PI3K/Akt/NF-κB signaling pathway. Conversely, DEC1 knockdown diminished these effects. Notably, treatment with the inhibitor LY294002 or the NF-κB inhibitor EVP4593 significantly reversed the IL-6-induced upregulation of DEC1, the increase in N-cadherin and vimentin, and the reduction in E-cadherin. These in vitro findings were further corroborated in vivo. Dec1 +/+ CRC mice exhibited elevated Il6 mRNA levels and enhanced tumor invasion compared with Dec1 -/- CRC mice. Dec1 deficiency increased E-cadherin and decreased N-cadherin and vimentin, accompanied by inhibition of the PI3K/Akt/NF-κB signaling pathway in mice. Taken together, our results demonstrate that DEC1 mediates IL-6-induced invasion and migration of CRC via activation of the PI3K/Akt/NF-κB signaling pathway.
Ischemic stroke stands as a principal driver of global mortality and permanent functional deficits. Notably, the clinical efficacy of current interventions is severely restricted by post-ischemic neuroinflammation. Cerebral ischemic injury prompts an inflammatory surge mediated by the cGAS-STING signaling cascade, a process initiated by the recognition of aberrantly localized cytosolic DNA. Three-prime repair exonuclease 1 (TREX1), a cytosolic DNA exonuclease, negatively regulates STING signaling; however, efficient delivery of TREX1 to the ischemic brain remains a major challenge. Here, we engineered mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) as carriers for TREX1 delivery. This was accomplished by using a palmitoylation signal-fused construct (PALM-TREX1), which enables efficient loading of the protein into extracellular vesicles. The C1C2 domains of lactadherin and the RGD-4C peptide were combined into a single recombinant fusion protein, enabling EV surface functionalization for enhanced ischemic targeting via phosphatidylserine interaction. In a mouse model of middle cerebral artery occlusion (MCAO), RGD-modified TREX1-loaded EVs (RGD-EV-TREX1) preferentially accumulated in ischemic regions, suppressed STING pathway activation, and reduced microglial activation and pro-inflammatory cytokine expression. The reduction in neuronal DNA damage and apoptosis ultimately facilitated improved neurological functional recovery, positioning RGD-EV-TREX1 as a promising cell-free therapeutic strategy for ischemic stroke.
Cardiac xenotransplantation (CXTx) has emerged as a potentially transformative solution to the global shortage of donor organs, driven by recent breakthroughs in genome-editing technologies. This review provides a comprehensive overview of the field, tracing its evolution from early experimental barriers to the current stage of early clinical translation. We summarize the significant strides made in the development of genetically engineered donor pigs, in which multi-gene modifications have effectively overcome the obstacle of hyperacute rejection. Despite these advances, long-term graft survival remains limited by complex immunological and physiological challenges, including acute humoral xenograft rejection, cellular immune responses, and coagulation dysregulation. Furthermore, this review critically analyzes data from recent milestones, including studies in brain-dead decedent models and the first genetically modified pig-to-human heart transplants. These clinical endeavors have exposed critical unresolved issues, particularly antibody-mediated rejection and the biosafety risks associated with porcine viruses. In conclusion, we discuss the key pathways for future progress, emphasizing the urgent need for optimized immunosuppressive regimens, rigorous viral surveillance, and standardized preclinical protocols to establish CXTx as a safe and durable clinical reality.
Chimeric antigen receptor T-cell (CAR-T) therapy represents a major advance in cellular immunotherapy and has demonstrated substantial clinical benefit in relapsed or refractory B-cell malignancies. However, autologous CAR-T therapy remains constrained by manufacturing complexity, high cost, variability in product quality, and treatment delays that may compromise outcomes in rapidly progressing disease. Allogeneic "off-the-shelf" CAR-T cell approaches have emerged as a potential strategy to address these limitations by enabling standardized manufacturing, rapid availability, and scalable production. Nevertheless, these theoretical advantages must be carefully balanced against significant challenges, including alloreactivity, immune rejection, complex genome engineering requirements, and regulatory constraints. This review provides a critical and balanced overview of the advantages and limitations of allogeneic CAR-T cell therapy, with a particular focus on applications in solid tumors. We discuss key biological barriers, including tumor microenvironment-mediated immunosuppression, and evaluate current engineering strategies aimed at enhancing efficacy, along with emerging clinical data. Collectively, while allogeneic CAR-T therapies hold considerable promise, substantial scientific, technical, and regulatory challenges must be addressed before their widespread clinical implementation.
Osteoarthritis (OA) is a degenerative joint disease that occurs frequently in middle-aged and elderly individuals, limiting joint function and causing disability with severe pain. However, available treatment options for inhibiting inflammation, preventing cartilage degradation, and relieving pain remain limited. Paeoniflorin, a monoterpene glycoside isolated from the Chinese medicine Paeonia lactiflora exhibits anti-inflammatory and immunomodulatory effects. In the present study, we investigated the efficacy and possible mechanisms of paeoniflorin in attenuating pain, inflammation, and cartilage degradation in a mouse model of OA. The analgesic effect of paeoniflorin was assessed by measuring mechanical allodynia with von Frey hairs. H&E staining was used to evaluate the structural integrity and inflammation of joint tissues. RAW264.7 cells were used to investigate the effects of paeoniflorin on related signaling pathways and on lipopolysaccharide (LPS)-induced inflammation by Western blotting. Paeoniflorin relieved mechanical allodynia and reduced cartilage degeneration in vivo. Moreover, paeoniflorin upregulated Gas6 expression, activated the Axl receptor, upregulated suppressor of cytokine signaling 3 (SOCS3) expression, inhibited MMP-9 expression, and decreased p38 phosphorylation in the joints. Cell experiments revealed that paeoniflorin regulated the Gas6-TAM pathway via the ERK signaling pathway and decreased MMP-9 expression and M1 polarization. Collectively, paeoniflorin may inhibit joint inflammation and relieve pain by upregulating the Gas6-TAM pathway and promoting macrophage M2 polarization, suggesting that it may serve as a potential therapeutic agent for osteoarthritis.
The infiltration of pro-inflammatory macrophages and the enzymatic degradation of the protective intra-islet heparan sulfate (HS) barrier are established pathological hallmarks of type 1 diabetes (T1D). While we previously identified myeloid-derived heparanase (HPSE) as the primary enzyme responsible for intra-islet HS cleavage, the transcriptional mechanisms driving its aberrant upregulation in macrophages remain unknown. By integrating single-cell RNA sequencing of T1D immune cells, we identified the transcription factor ETS proto-oncogene 1 (ETS1) as a key upstream regulator of Hpse in T1D-specific macrophages. Mechanistically, Cleavage Under Targets and Tagmentation (CUT&Tag) and luciferase reporter assays confirmed that ETS1 directly binds to the Hpse promoter and activates its expression in macrophages. In vivo, myeloid-specific Ets1 knockout ( Ets1-mKO) mice exhibited profound resistance to multiple low-dose streptozotocin (MLD-STZ)-induced T1D insulitis. This protection was driven by the marked suppression of myeloid HPSE expression, which preserved intra-islet HS levels, reduced inflammatory cell infiltration, and enhanced β-cell survival compared with that in wild-type littermates. In conclusion, our findings define a previously unrecognized ETS1-HPSE-HS signaling axis that is involved in macrophage-mediated islet damage in T1D. We demonstrate that ETS1 is a crucial driver of the enzymatic breakdown of the islet basement membrane and the subsequent progression of insulitis, suggesting that targeting the ETS1-mediated transcriptional activation of HPSE offers a novel therapeutic strategy to safeguard the islet microenvironment and slow the progression of T1D.
The four original articles in this special section explore diverse aspects of cancer biology,ranging from molecular mechanisms and tumor heterogeneity to behavioral risk prediction,thereby highlighting the power of integrating experimental biology with computational approaches.
Helicobacter pylori ( H. pylori) infection is a major risk factor for gastric cancer, though the immune microenvironmental factors driving preneoplastic transformation remain unclear. In this study, single-cell RNA sequencing was used to characterize macrophage subsets and identify key genes associated with H. pylori-induced gastric lesions. Molecular, cellular, and in vivo assays were conducted to determine the mechanisms regulating the expression of a disintegrin and metalloproteinase-like decysin 1 ( ADAMDEC1) and its downstream effects. M2 macrophages were significantly enriched in H. pylori-positive gastritis and metaplasia, and ADAMDEC1 showed progressive increase in expression during lesion progression. H. pylori promoted M2 polarization and induced ADAMDEC1 transcription via signal transducer and activator of transcription 3 (STAT3) binding to its promoter. Functionally, M2-derived ADAMDEC1 increased soluble epidermal growth factor levels, thereby activating the epidermal growth factor receptor (EGFR)-extracellular signal-regulated kinase 1/2 (ERK1/2) signaling pathway in epithelial cells, which led to upregulation of C-C motif chemokine ligand 20 (CCL20) expression, enhanced proliferation, and impaired genomic stability. The present study reveals a potential H. pylori-driven oncogenic pathway involving the M2 macrophage-ADAMDEC1-EGF/EGFR/ERK axis, establishing ADAMDEC1 as a potential target for early intervention in H. pylori-associated gastric cancer.
Obesity-related cardiomyopathy (OCM) is characterized by pathological cardiac remodeling and progressive functional decline, often accompanied by mitochondrial dysfunction, particularly aberrant mitophagy. The role of the core circadian gene brain and muscle ARNT-like protein 1 ( Bmal1) in OCM remains unclear. In this study, we employed a high-fat diet (HFD)-induced OCM mouse model, a cardiomyocyte-specific Bmal1 knockout ( Bmal1 CMKO) model, and a palmitic acid (PA)-induced H9c2 cardiomyocyte injury model to investigate the function of Bmal1. In vivo, BMAL1 expression was reduced in hearts of HFD mice; HFD- Bmal1 CMKO mice exhibited exacerbated myocardial hypertrophy, fibrosis, functional impairment, and apoptosis, accompanied by increased expression of the mitophagy-related proteins PINK1, Parkin, and LC3-II. In vitro, PA exposure decreased BMAL1 expression, disrupted mitochondrial membrane potential, increased reactive oxygen species generation, and induced excessive mitophagy; these effects were aggravated by Bmal1 silencing and attenuated by Bmal1 overexpression, which also improved cell viability. Collectively, these findings indicate that Bmal1 plays a protective role in OCM, and its downregulation may be a key contributor to obesity-induced cardiac remodeling and dysfunction. Mechanistically, BMAL1 downregulation was accompanied by activation of the PINK1/Parkin signaling and enhanced mitophagy under lipid stress. By restraining excessive mitophagy and preserving mitochondrial function and metabolic homeostasis, Bmal1 and its associated pathways may represent promising therapeutic targets for OCM.
This study evaluated a multi-parameter serological model for the auxiliary diagnosis of aortic aneurysm (AA). Clinical data and serological indicators were collected from 1397 AA patients and 2000 healthy controls recruited from the Second Affiliated Hospital of Harbin Medical University (June 2021-June 2025). A comprehensive diagnostic model was developed based on serum biomarkers. Significant differences were observed between AA patients and healthy controls in blood pressure, body mass index (BMI), inflammatory markers ( e.g., white blood cell count, and neutrophil count), hepatic and renal function indicators ( e.g., urea, total bilirubin, and alkaline phosphatase), and nutritional parameters ( e.g., albumin, total protein, and hemoglobin) ( P < 0.05). Multivariate analysis identified BMI, systolic blood pressure, urea, and neutrophil count as independent risk factors (OR > 1), while lymphocyte count, albumin, and serum potassium were protective factors (OR < 1). Twenty-three key variables were further selected by least absolute shrinkage and selection operator (LASSO) regression. The constructed comprehensive RiskScore showed a highly significant difference between the AA group and the control group (-16.27 ± 4.69 vs. -21.36 ± 1.46, P < 0.001), with the model demonstrating excellent discriminatory performance (concordance index [C-index] > 0.9). The results indicate that a multi-parameter serological diagnostic model distinguishes between AA patients and healthy controls, demonstrating strong discriminatory ability and potential uitility for auxiliary diagnosis of AA.
The longitudinal associations between emotional and behavioral problems and Internet addiction, including how transitions between problem patterns influence risk, remain underexplored. This study investigated these associations using longitudinal data from 5123 primary and secondary school students in Jiangsu Province, China. Latent class analysis and latent transition analysis were applied to identify patterns and transitions in emotional and behavioral problems, and generalized estimating equations were used to examine their associations with subsequent Internet addiction. Emotional symptoms, conduct problems, hyperactivity/inattention, peer problems, and prosocial behavior were all significantly associated with Internet addiction. Three latent classes were identified: high difficulties, social difficulties, and low symptoms. The high difficulties group exhibited the greatest risk of Internet addiction. Compared with those maintaining stable classes, participants transitioning from the low symptoms class to the social difficulties or high difficulties classes showed elevated risks of Internet addiction (odds ratio [OR] = 2.37, 95% confidence interval [CI] = 1.58-3.55; OR = 5.62, 95% CI = 3.23-9.78). Conversely, those moving from the high difficulties class to the social difficulties or low symptoms classes had reduced risks (OR = 0.58, 95% CI = 0.36-0.94; OR = 0.28, 95% CI = 0.13-0.60). These findings suggest that both emotional and behavioral problems and their dynamic transitions are closely linked to Internet addiction risk in children and adolescents. Early identification, continuous monitoring, and timely interventions to promote positive transitions are critical for effective prevention.
Hepatic accumulation of bile acids (BAs) contributes to cholestasis-induced liver injury and fibrosis. Our previous studies have shown that Lactobacillus murinus ( L. murinus) alleviates liver fibrosis in primary sclerosing cholangitis (PSC) by modulating bile acid metabolism. In this study, we found that L. murinus intervention alleviated hepatic taurocholic acid (TCA) accumulation, cholestasis, and liver fibrosis in PSC mice. TCA was shown to promote liver fibrosis in PSC mice by enhancing M2 macrophage-mediated secretion of transforming growth factor beta 1 (TGF-β1). Macrophage depletion in PSC mice reduced the proportion of M2 macrophages and TGF-β1 cytokine levels, alleviating liver fibrosis. L. murinus exhibited bile salt hydrolase (BSH) activity, preferentially hydrolyzing glycine-conjugated bile acids. Furthermore, L. murinus reduced TCA reabsorption into the portal circulation by suppressing farnesoid X receptor ( Fxr) gene expression in the ileum, thereby alleviating hepatic TCA accumulation and mitigating liver fibrosis in PSC mice. These findings suggest that L. murinus mitigates PSC-associated liver fibrosis, which is associated with reduced TCA levels and regulation of M2 macrophage polarization, offering a potential therapeutic strategy for PSC.
Heavy metal contamination has become an emerging global health concern, yet the promoting effects of mixed heavy metal exposure on colorectal cancer risk remains poorly understood. In this study, plasma concentrations of 25 heavy metals were obtained from an in-house elemental database comprising both colorectal cancer patients and healthy controls, and exposure concentrations were then defined based on these human plasma detection data and relevant public datasets. Machine learning algorithms and mixture exposure models were applied to identify key heavy metals associated with colorectal cancer risk. RNA-seq datasets were integrated to screen for genes exhibiting heavy metal-tumor-specific characteristics. A total of 270379 cells from 95 samples were analyzed to identify cell subsets sensitive to mixed heavy metal exposure. Molecular and cellular experiments were performed to validate the underlying mechanisms. We first identified stannum (Sn), antimony (Sb), tantalum (Ta), and thallium (Tl) as a mixture of heavy metal highly associated with colorectal cancer risk. ESM1, SLC7A5 and GRIA4 were further identified as key genes linked to colorectal tumors under conditions of mixed heavy metal exposure. Based on scRNA-seq analyses, endothelial tip cells were first identified as exhibiting heightened sensitivity to mixed heavy metal exposure. Mechanistically, acute mixed heavy metal exposure upregulated VEGFA expression in colorectal tumor epithelial cells. Co-culture experiments demonstrated enhanced epithelial-endothelial communication through the VEGFA-FLT1 axis, accompanied by increased expression of ESM1 and ANGPT2 in endothelial cells. Collectively, these findings suggest that mixed heavy metal exposure promotes angiogenesis by enhancing the interaction between colorectal tumor epithelial and endothelial cells, thereby increasing the risk of colorectal cancer. This study provides a scientific basis for improving strategies to control heavy metal contamination.
Cytochrome P450 CYP3A (CYP3A) is among the most abundant hepatic cytochrome P450 subfamilies and also mediates the metabolism and toxicity of xenobiotics. Previous studies have reported that CYP3A is also expressed in the testis; however, its role and molecular mechanism in mediating male reproductive damage remain unclear. In this study, through in vitro and in vivo experiments, we demonstrated the role of CYP3A in 2,2',4,4'-tetrabromodiphenyl ether (BDE47)-induced reproductive toxicity. The results showed that BDE47 induced CYP3A expression in mouse testes, leading to oxidative stress and ferroptosis through excessive reactive oxygen species (ROS) and ferrous iron (Fe 2+) overload, which was demonstrated by CYP3A overexpression or knockdown experiments in GC-2 cells. Mechanistically, in addition to direct ROS generation during metabolic processing, ferritinophagy contributed to intracellular Fe 2+ accumulation. Specifically, BDE47-induced ROS was associated with reduced N6-methyladenosine (m 6A) modification of Atg12 mRNA and increased autophagy-related (ATG12) expression, thereby promoting ferritin heavy chain 1 (FTH1) degradation and subsequent Fe 2+ overload. These results were further validated by experiments using hydrogen peroxide or antioxidants in GC-2 cells, as well as by Atg12 haploinsufficiency in mice. Our findings demonstrate that CYP3A plays a critical role in male reproductive toxicity induced by BDE47.
