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.
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.
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.
Growing evidence highlights the gut, reproductive tract, and endometrial microbiota as important functional contributors in the pathogenesis of endometriosis (EM). Studies have revealed a characteristic microbial imbalance in patients with EM, marked by a reduced abundance of beneficial bacteria and an enrichment of opportunistic pathogens. These microbial communities are thought to influence disease progression primarily through metabolic activity, as demonstrated by metabolomic studies showing their capacity to modulate host immune and endocrine responses. This imbalance may contribute to several key metabolic disturbances, including decreased levels of short-chain fatty acids, particularly butyrate; a shift in tryptophan metabolism toward the kynurenine pathway; elevated β-glucuronidase activity; increased lipopolysaccharide production; and altered secondary bile acid profiles. Functionally, these metabolic alterations are thought to contribute to EM by disrupting immune homeostasis, enhancing estrogen signaling, and driving systemic inflammation, thereby creating a permissive microenvironment for ectopic lesion growth and invasion. Targeted interventions, such as probiotics, high-fiber dietary strategies, fecal microbiota transplantation, and selective modulation of microbial or host metabolic enzymes, are emerging as promising non-hormonal therapeutic approaches. Nonetheless, further studies incorporating longitudinal cohort designs and integrative multi-omics approaches are essential to establish causality and facilitate the development of precise diagnostic and personalized treatment strategies.
Bisphenol A (BPA), a widely encountered environmental endocrine-disrupting chemical, has been linked to an increased risk of congenital heart defects in epidemiological and experimental studies. To investigate its impact in a human-relevant system, we used a human cardiac organoid (hCO) model derived from human embryonic stem cells and exposed hCOs to BPA concentrations ranging from environmentally relevant to higher experimental levels (0.1-10 μmol/L) throughout differentiation. BPA exposure impaired cardiac contractile function and disrupted endothelial network formation, with more pronounced functional effects at higher concentrations. BPA exposure also induced apoptosis in hCOs. Subsequently, transcriptomic profiling revealed dysregulation of genes associated with cardiac rhythm and developmental signaling in BPA-treated hCOs. Among the concentrations tested, 0.5 μmol/L was the lowest concentration associated with a significant reduction in CD31 fluorescence intensity. This research provides in vitro evidence that BPA can perturb key processes in early human heart development, highlighting the necessity for further mechanistic and in vivo studies to clarify exposure relevance and its potential implications for fetal heart health.
Dear Editor, Age-related macular degeneration (AMD) is a leading cause of visual impairment and vision loss among older adults worldwide.
In China, gestational diabetes mellitus (GDM) is typically diagnosed using the International Association of Diabetes and Pregnancy Study Groups (IADPSG) criteria. This study retrospectively analyzed 19152 pregnant women who underwent an oral glucose tolerance test between 2015 and 2021, comparing the IADPSG and the National Institute for Health and Care Excellence (NICE) criteria. GDM prevalence was 20.39% (IADPSG) and 23.67% (NICE, P < 0.01), with a moderate diagnostic agreement (κ = 0.59; P < 0.01). Compared with women having normal glucose tolerance (NGT group), women diagnosed with GDM by either the IADPSG (GDM-I) or NICE (GDM-N) criteria had significantly higher risks of adverse outcomes (including gestational hypertension, preeclampsia, preterm delivery, macrosomia, low birth weight, and neonatal jaundice). However, those diagnosed by NICE but not by IADPSG (GDM-NI) showed no significant increase in adverse maternal or neonatal outcomes except for neonatal jaundice. Furthermore, the GDM-NI group had significantly lower rates of several adverse outcomes compared with the GDM-I group. These findings suggest that, under current clinical management where women diagnosed by IADPSG received treatment while those diagnosed only by NICE did not, the IADPSG criteria classified fewer women as having GDM but identified a group with a higher observed risk of several adverse pregnancy outcomes than did the NICE criteria.
Medulloblastoma(MB)is the most common malignant tumor of the cerebellum in children.The Sonic Hedgehog(SHH)subgroup of MB(SHH-MB)is driven by aberrant activation of the SHH pathway;however,mutations in genes related to this pathway are relatively rare,posing challenges for therapeutic development.Glypican-6(GPC6),a heparan sulfate proteoglycan,is highly expressed in SHH-MB.In this study,we demonstrate the synchronous expression of GPC6 with GLI family zinc finger 1(GLI1)in both the developing cerebellum and medulloblastoma.GPC6 promotes cell proliferation,migration,and invasion in SHH-MB cell lines(DAOY and ONS-76).Consistently,GPC6 enhances the SHH pathway activity by upregulating GLI1 expression,supports ciliogenesis essential for signal transduction,and facilitates SHH ligand expression via extracellular vesicles.These findings suggest that GPC6 acts as a key regulator of SHH signaling and represents a potential therapeutic target in SHH-MB.
The genetic landscape of male infertility is highly complex. It is estimated that at least 1000-2000 genes are involved in human and mouse infertility. Although functional analyses have been performed on hundreds of genes, many others still have unknown functions. Generating gene-editing mice is a powerful tool for exploring whether a given gene is essential for male reproduction in vivo. In the current study, we investigated the function of six genes, Efcab7, Tekt3, Mlf1, Rp1l1, Agbl2, and Tmsb15a, using the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system. Individual disruption of these six genes did not cause overt male sterility in mice under standard breeding conditions. Phenotypic analyses at 8-10 weeks of age generally showed that most reproductive parameters were comparable to those of wild-type controls; however, we observed significant sperm motility alterations in Tekt3-mutant mice (decreased motility) and Rp1l1-mutant mice (increased motility), as well as flagellar abnormalities in Tekt3 mutants. In summary, these findings suggest that complete or near-complete disruption of the tested murine alleles does not cause overt male infertility, providing useful functional evidence for interpretation of candidate variants while not excluding human-specific or allele-specific effects.
While infections have been implicated in endometriosis pathogenesis, the role of human papillomavirus (HPV) remains unclear. This study combined a meta-analysis of seven studies with a case-control study ( n = 432 surgically treated patients) to evaluate the association of HPV infection with endometriosis risk and fertility outcomes. The meta-analysis showed no significant association between HPV infection and endometriosis risk, either for any HPV subtype (pooled odds ratio [OR] = 2.60, 95% confidence interval [CI]: 0.28-23.87) or high-risk subtypes (OR = 1.68, 95% CI: 0.49-5.75). Notably, HPV prevalence was higher in patients with endometriosis (46% overall; 36% for high-risk subtypes) than in the general population. Evidence regarding the association between HPV infection and fertility outcomes remains limited, with only two eligible studies identified. Our case-control analysis revealed that HPV-positive patients had significantly reduced postoperative live birth rates compared with HPV-negative counterparts (10.6% vs. 21.0%, P < 0.05). These findings suggest that HPV infection is unlikely to be a major risk factor for endometriosis development, but it may be associated with poorer postsurgical fertility outcomes. The dual-method approach strengthens the evidence for HPV's clinical impact on reproductive prognosis in patients with existing endometriosis.
Ocular diseases,including corneal disease,glaucoma,age-related macular degeneration(AMD),diabetic retinopathy,and retinopathy of prematurity,can significantly impair vision and reduce quality of life.Because degenerated cells in these diseases are unable to regenerate,treatments for these conditions have limited efficacy.Stem cell therapies are revolutionizing the treatment of degenerative eye conditions,enabling structural and functional restoration through mechanisms such as cell replacement and paracrine signaling.This review examines advances in stem cell therapy for ocular diseases,from preclinical studies to early clinical trials,focusing on various types of stem cells,including embryonic stem cells,induced pluripotent stem cells(iPSCs),and mesenchymal stem cells.Significant progress has been made with iPSC-derived retinal pigment epithelial cell transplantation in AMD treatment,showing cell survival in trials,and with mesenchymal stem cells for corneal repair through anti-inflammatory effects.Challenges remain,such as controlling differentiation to prevent tumorigenesis,managing immune rejection,and ensuring manufacturing processes that comply with Good Manufacturing Practice standards.By integrating mechanistic insights with translational strategies,this review outlines pathways to optimize stem cell therapies for previously intractable ocular diseases.
Non-obstructive azoospermia (NOA), characterized by impaired spermatogenesis and the complete absence of sperm in the ejaculate, represents one of the most severe forms of male infertility. Current approaches for definitive histological characterization and sperm retrieval often require invasive testicular procedures, underscoring the need for reliable, non-invasive methods for preoperative stratification. In the present study, we performed untargeted metabolomic profiling of human seminal plasma to identify biomarker panels capable of stratifying azoospermia subtypes through a stepwise approach. We identified distinct metabolite biomarker panels comparing NOA vs. obstructive azoospermia (OA), Sertoli cell-only syndrome vs. other NOA subtypes, and hypospermatogenesis vs. maturation arrest. Additionally, cross-species comparative analysis with high-resolution metabolomic data from purified mouse testicular cell populations implicated these biomarkers in specific germ cell stages and metabolic transitions during spermatogenesis. These findings provide a preliminary molecular framework for the non-invasive classification of azoospermia subtypes and warrant validation in larger independent cohorts.
Dear Editor, In addition to the classic barriers and challenges encountered during the design,conduct,and reporting of research,researchers and scholars now face a growing dilemma:selecting the most appropriate journal in which to publish their work.
Dear Editor, Macrovascular and microvascular diseases are the most frequent complications of type 2 diabetes mellitus (T2DM),with coronary artery disease (CAD)being particularly noteworthy[1].
As the most prevalent cause of death worldwide, ischemic stroke urgently requires innovative therapeutic strategies. The present study demonstrated the therapeutic potential of human umbilical cord-derived mesenchymal stem cell-derived exosomes (hUMSC-Exos) in ameliorating hypoxia-induced cerebrovascular endothelial dysfunction through modulation of the AMPK/NLRP3 signaling pathway. Bioinformatics analysis of DisGeNET and exosomal cargo databases revealed 283 overlapping cerebral ischemia-related genes, implicating hUMSC-Exos in inflammatory regulation. In vitro experiments showed that hUMSC-Exos rescued oxygen-glucose deprivation (OGD)-induced endothelial dysfunction in bEnd.3 mouse brain endothelial cells, restoring viability, migration, and mitochondrial integrity. Mechanistically, hUMSC-Exos reversed OGD-induced AMPK inactivation while suppressing NLRP3 inflammasome activation, caspase-1 cleavage, and gasdermin D (GSDMD)-mediated pyroptosis. Molecular docking revealed that DL-3-n-butylphthalide acts as a dual-target ligand for AMPK/NLRP3, synergizing with hUMSC-Exos to enhance endothelial protection. In vivo, combined therapy in the transient middle cerebral artery occlusion mouse model reduced cerebral infarction and improved neurological outcomes, accompanied by NLRP3/GSDMD downregulation and hippocampal neuron preservation. These findings establish hUMSC-Exos as regulators of AMPK/NLRP3-mediated pyroptosis and propose a translatable combinatorial regimen for ischemic stroke therapy.
Dear Editor, Psoriasis is an inflammatory skin disease linked to many other medical conditions[1]. Research indicates that psoriasis patients face an increased risk of developing comorbid inflammatory diseases,such as arthritis and chronic obstructive pulmonary disease,which includes chronic bronchitis and emphysema[2].
Lung cancer in smokers (LCIS) and lung cancer in never-smokers (LCINS) are different entities with distinct molecular features. However, their cellular heterogeneity still requires further investigation. Through an integrated analysis of single-cell RNA sequencing and bulk sequencing data, we identified cell subpopulations associated with smoking and non-smoking patients. Subsequent transcriptomic analyses were performed to elucidate differences in cellular functions and the tumor microenvironment. We observed that smoking-associated cancer cells exhibited a higher degree of aggressiveness, which may correlate with an adverse prognosis in smoking patients. Additionally, immunosuppressive CXCL10 + macrophages may contribute to tumorigenesis in smokers, and the immunoregulatory LGALS9-HAVCR2 axis could be a potential immunotherapeutic target. In non-smokers, the inflammatory microenvironment may be involved in tumor development. Moreover, the reduced anti-tumor cytotoxicity may be associated with their suboptimal immunotherapeutic response. Our study uncovered differences in oncogenic and immune escape mechanisms between LCIS and LCINS patients and suggests potential immunotherapeutic strategies.
While a healthy lifestyle is known to reduce the risk of stroke, the extent to which blood pressure (BP) mediates this association remains unclear. The present study aimed to quantify the mediating role of BP in the association between combined lifestyle factors and stroke incidence. Using data from 51929 participants free of major cardiovascular diseases or cancer at baseline, we employed structural equation modeling to assess the mediating effects of systolic (SBP) and diastolic (DBP) blood pressure. During the follow-up, 2811 incident stroke cases were identified. A healthy lifestyle was significantly associated with a reduced risk of stroke, with SBP mediating 44.70% ( β = -0.0014, 95% confidence interval [CI]: -0.0016 to -0.0012) and DBP mediating 37.81% ( β = -0.0012, 95% CI: -0.0015 to -0.0009) of this association. The mediating effects were attenuated but remained significant for ischemic stroke (SBP: 33.21%; DBP: 27.24%). In conclusion, approximately two-fifths of the protective association between a healthy lifestyle and stroke may be mediated by BP. These findings suggest that BP control may serve as an important early indicator for evaluating the effectiveness of lifestyle interventions in reducing stroke risk.
PIWI-interacting RNAs(piRNAs)are a class of noncoding RNAs primarily found in germ cells.While piRNAs are known to be involved in various cancers,their specific roles in colorectal cancer(CRC)remain unclear.To elucidate the role of piRNAs in CRC,we first analyzed their expression characteristics by sequencing 10 pairs of tumor and adjacent normal tissues.Subsequently,differentially expressed piRNAs were identified through a two-stage reverse transcription-quantitative PCR(RT-qPCR)validation using 20 and 114 pairs of samples.Subcellular localization was assessed through nucleoplasmic separation and immunofluorescence staining assays.RNA pull-down mass spectrometry was employed to identify piRNA-interacting proteins.We identified piR-61298 as a piRNA significantly upregulated in CRC.Functional assays showed that piR-61298 promoted cell proliferation and migration,inhibited apoptosis,and promoted tumor growth.Mechanistically,piR-61298 bound to ubiquitin-specific peptidase 10(USP10)in the cytoplasm,impairing its deubiquitinating activity toward p53,thereby leading to p53 ubiquitination and degradation.These findings suggest that piR-61298 plays a critical role in CRC progression by disrupting the USP10-p53 axis.Collectively,the current study highlights piR-61298 as a potential therapeutic target,offering a novel approach for CRC treatment by targeting piRNA-mediated regulation.
Diabetic retinopathy (DR), a common complication of diabetes, is characterized by retinal angiogenesis and inflammation. The role of hepatoma-derived growth factor (HDGF) in mediating inflammation during DR remains unclear. We measured HDGF levels in the aqueous humor and found that HDGF was increased in DR but decreased after anti-angiogenesis treatment. Using public single-cell RNA sequencing datasets, we found that elevated HDGF in DR was mainly produced by Müller cells and targeted microglia. Additionally, integrin beta 2 ( Itgb2), a target gene of HDGF that induces microglial activation, was significantly upregulated in DR. To verify these results, we performed enzyme-linked immunosorbent assays, quantitative reverse transcription-PCR, Western blotting, and fluorescence immunostaining in cultured Müller and microglial cells treated with HDGF or anti-HDGF, as well as in DR mice receiving intravitreal injections of HDGF or its antibody. Exogenous HDGF further promoted microglial activation, migration, and secretion of pro-inflammatory cytokines, while neutralization of HDGF suppressed these effects caused by high glucose. Furthermore, the HDGF receptor nucleolin was overexpressed in microglia under high glucose stimulation. Therefore, blocking HDGF from Müller cells in DR reduced the excessive inflammatory response in microglia, highlighting HDGF as a potential therapeutic target.
