CYP17A1 gene (Cytochrome P45017A1) plays a significant role in the steroidogenic pathway that synthesizes sex steroids and glucocorticoids. In Cytochrome P45017A1, the heme-binding domain region is essential for its catalytic function to synthesize steroids. Mutations in the CYP17A1 gene have been associated with several pathological conditions, including congenital adrenal hyperplasia, Polycystic Ovary Syndrome (PCOS), now referred to as Polyendocrine Metabolic Ovarian Syndrome (PMOS), and certain types of cancers. This study aimed to identify deleterious missense mutations in CYP17A1 using in silico tools, molecular dynamics simulations, and genotyping to detect both rare and common variants. In silico analyses identified 22 pathogenic mutations using sequence- and structure-based tools. Furthermore, the mutations in the heme-binding domain (F453S, R449C) were characterized using molecular dynamics simulations. Results of Molecular dynamics simulations show that CYP17A1 mutants exhibit significant structural changes, particularly in the heme and steroid-binding regions, which weaken heme coordination and alter heme affinity. Based on the high minor allele frequency (MAF) of 0.04
Advances in cancer therapy have markedly improved patient survival, thereby increasing the need for effective fertility preservation strategies. Chemotherapeutic agents commonly induce apoptosis in rapidly proliferating cells, including spermatogonia, resulting in long-term testicular damage. Y-27632, a Rho-associated coiled-coil–containing protein kinase (ROCK) inhibitor, has been shown to suppress apoptosis in various stem cell populations by modulating Rho GTPase–dependent signaling pathways. This study aimed to evaluate the protective effects of Y-27632 against busulfan-induced testicular injury in a murine model. Eighteen 4-week-old male C57BL/6 mice were randomly assigned to three groups: control (no treatment), busulfan (40 mg/kg), and busulfan plus Y-27632 (40 mg/kg busulfan + 5 mg/kg Y-27632). Seventy days after treatment, sperm parameters (count, motility, viability, and plasma membrane integrity) and testicular outcomes, including seminiferous tubule diameter, germinal epithelium thickness, tubular differentiation index, spermatogenesis index, Johnsen score, apoptotic gene expression, malondialdehyde levels, total antioxidant capacity, and TUNEL-positive cells, were assessed. Y-27632 administration significantly improved seminiferous tubule diameter and germinal epithelium thickness and increased tubular differentiation and spermatogenesis indices and Johnsen scores compared with the busulfan group. In addition, Y-27632 treatment was associated with reduced ERK gene expression and a lower number of TUNEL-positive cells. However, sperm parameters and testicular weight remained significantly reduced compared with controls, and no significant differences were observed in lipid peroxidation, total antioxidant capacity, or sperm DNA integrity among groups. In conclusion, Y-27632 partially attenuated busulfan-induced testicular damage by improving histological indices of spermatogenesis and reducing apoptosis, suggesting that ROCK inhibition may represent a potential adjunct strategy for fertility preservation following gonadotoxic cancer therapies.
Chronic endometritis (CE) is a persistent inflammatory condition of the endometrium characterized by stromal plasma cell infiltration and is increasingly recognized in women with infertility, recurrent implantation failure (RIF), and recurrent pregnancy loss (RPL). Although traditionally considered a localized infectious process, heterogeneous treatment responses and recurrence suggest that broader immunological mechanisms may contribute in selected patients. This structured narrative review synthesizes clinical and mechanistic evidence relevant to a possible gut–immune–endometrial axis. A structured PubMed/MEDLINE search was conducted from database inception to December 31, 2025, and reference lists of relevant reviews and key primary studies were screened. Evidence was synthesized thematically across clinical, immunological, and translational domains. CE is associated with infertility and RPL, whereas findings in RIF populations are heterogeneous and strongly dependent on diagnostic criteria. CE is characterized by plasma cell infiltration, pro-inflammatory signaling, and activation of pathways including toll-like receptor 4 (TLR4)–nuclear factor kappa B (NF-κB). Separately, intestinal dysbiosis and barrier dysfunction may permit systemic translocation of microbial products such as lipopolysaccharide, thereby promoting low-grade inflammatory signaling. However, no direct human study has yet linked gut dysbiosis or endotoxemia to histologically confirmed CE within the same cohort. The proposed axis should therefore be regarded as a hypothesis-generating framework rather than an established causal pathway. CE is biologically heterogeneous and may arise from infectious, local microbial, immune-mediated, or systemic-inflammatory mechanisms that can overlap. Reverse causality and shared confounding factors are also plausible. At present, the model does not justify routine microbiome testing or gut-directed treatment for CE, but it defines priorities for prospective studies integrating microbiome profiling, systemic immune markers, and standardized endometrial phenotyping.
Obesity induces male reproductive dysfunction. Mitochondrial dysfunction ranks among the mechanisms driving functional impairment. Notably, Mitochondrial NADH dehydrogenase subunit 4 (MTND4) exerts a pivotal role in sustaining the function of the mitochondrial electron transport chain and cellular energy metabolism. However, its molecular mechanism in obesity-associated seminal vesicle injury remains incompletely elucidated. In vivo and in vitro models of high-fat diet (HFD) were established, and the body weight changes of mice were recorded. Corresponding assay kits were applied to detect serum testosterone, total cholesterol (TC), and triglycerides (TG) levels, with changes in metabolic phenotypes analyzed. Hematoxylin-eosin staining, computer-aided sperm analysis system, qRT-PCR, Western blot, terminal deoxynucleotidyl transferase dUTP nick-end labeling, reactive oxygen species (ROS) detection, JC-1 staining, and superoxide dismutase/malondialdehyde assay kits were employed to evaluate seminal vesicle tissue structure, sperm function, cell viability, apoptosis, oxidative stress, and mitochondrial function, etc., respectively. The transcription factors upstream of MTND4 were forecast via the JASPAR database. The transcriptional regulation of MTND4 by STAT3 was verified via chromatin immunoprecipitation and dual-luciferase reporter assays. Finally, a STAT3-knockdown and MTND4-overexpression co-transfected HFD cell model was constructed to probe the effects of the STAT3/MTND4 on seminal vesicle injury and sperm motility. HFD induced obesity in mice, as evidenced by increased body weight, decreased serum testosterone level, and elevated levels of TC and TG. Concomitant manifestations included disorganized seminal vesicle tissue structure, epithelial exfoliation, impaired secretory function, together with marked reductions in sperm motility, progressive motility, and concentration. Meanwhile, the expression of MTND4 was markedly upregulated in mice with HFD. Overexpression of MTND4 promoted the accumulation of ROS, decreased mitochondrial membrane potential, and exacerbated oxidative stress, resulting in reduced motility and enhanced apoptosis of seminal vesicle cells. Furthermore, STAT3 transcriptionally activated MTND4; HFD induced a decrease in seminal vesicle cell viability via upregulating STAT3 to activate MTND4 transcription. HFD activates MTND4 transcription via upregulating STAT3, resulting in ROS production and mitochondrial dysfunction, which ultimately mediates obesity-related seminal vesicle injury and sperm dysfunction. The STAT3/MTND4 axis may be a novel target for regulating male reproductive health.
Accurate identification of ovulation remains challenging due to pronounced inter- and intra-individual variability of reproductive hormones. Volatile organic compounds (VOCs) in exhaled breath may vary across the menstrual cycle and offer a non-invasive complementary approach to menstrual cycle assessment. In this pilot study, exhaled breath VOCs were measured using proton transfer reaction time-of-flight mass spectrometry in five healthy women across follicular, ovulatory, and luteal phases of a natural menstrual cycle. The VOC profiles were compared between ovulatory and non-ovulatory measurements, and correlations with serum hormone concentrations of estradiol (E2), luteinizing hormone (LH), and progesterone (P) were explored. A significant difference between ovulatory and non-ovulatory measurements was observed for the ion m/z 41.04 (p = 0.021). Non-significant trends toward higher concentrations during ovulation were detected for isoprene-related signals. Breath VOCs exhibit phase-dependent alterations across the menstrual cycle. The observed increase in the m/z 41.04 signal during ovulation may reflect integrated metabolic adaptations associated with endocrine changes rather than direct surrogates of circulating hormone concentrations.
