Reproductive Biology and Endocrinology最新文献

Objective: This study aimed to investigate the microbiome profile in the cystic fluid of ovarian endometrioma and explore its association with the microbial communities present in the lower and upper reproductive tracts.

Design: A microbial analysis was conducted across multiple compartments of the reproductive tract in patients diagnosed with ovarian endometrioma.

Subjects: Sixteen female patients aged 25-43 years (mean age: 31.56 years) who underwent laparoscopic surgery for ovarian endometrioma at the First Hospital of Putian City between April 2023 and February 2024 were enrolled in this study.

Main outcome measures: 16S rDNA sequencing was employed to characterize the microbiome of ovarian endometrioma and assess its correlations with clinical symptoms, inflammatory markers, and serum CA125 levels RESULTS: Microbial communities were detected in the posterior vaginal fornix, endometrial fluid, peritoneal fluid, and cystic fluid, exhibiting distinct compositional profiles. Community diversity significantly increased along the anatomical gradient from the posterior vaginal fornix to endometrial fluid, peritoneal fluid, and cystic fluid, with the highest microbial diversity observed in the cystic fluid. Lactobacillus was the predominant genus in the posterior vaginal fornix, whereas Escherichia-Shigella was most abundant in endometrial fluid samples. Hydrogenophaga and Brevundimonas were the dominant taxa in both peritoneal and cystic fluids. Notably, the microbial composition of peritoneal fluid showed the greatest similarity to that of cystic fluid, and functional prediction analyses indicated largely overlapping biological functions between these two sites. Furthermore, Spearman correlation analysis revealed significant associations between specific microbial taxa and certain clinical manifestations or inflammatory factors.

Conclusion: This study demonstrates the presence of a unique and highly diverse microbiome within the cystic fluid of ovarian endometrioma. The site-specific microbial profiles and their correlations with clinical parameters suggest a potential role of microbiota in disease pathogenesis through inflammatory and metabolic mechanisms. These findings contribute novel insights that may inform future strategies for the prevention, diagnosis, and treatment of ovarian endometrioma.

Background: Non-steroidal anti-inflammatory drugs (NSAIDs) are commonly used to alleviate pain associated with endometriosis (EMS), yet the impact of their long-term use on disease progression remains unclear. This study investigates the dual role of NSAIDs in EMS pathogenesis using network toxicology and Mendelian randomization (MR).

Methods: The toxicity and ADMET profiles of nine NSAIDs were screened using ProTox 3.0 and ADMETlab 2.0. Potential drug targets were predicted using PharmMapper, STITCH, and SwissTargetPrediction, while EMS-related targets were retrieved from GeneCards, OMIM, and CTD databases. For the MR analysis, cis-eQTLs for whole blood tissue from GTEx v8 served as instrumental variables, based on the inflammatory nature of endometriosis. Outcome data were from an independent GWAS summary dataset (19,588 cases, 213,669 controls). Our analysis adhered to MR independence assumptions, ensuring no sample overlap between exposure and outcome data. Functional enrichment and molecular docking explored the underlying mechanisms.

Results: By integrating drug targets with disease genes, we first identified 463 overlapping targets. We revealed that EPHB4 is a core hub mediating the potential "risk-promoting" effects of nearly all NSAIDs, with its functions enriched in key pathological processes such as angiogenesis. Molecular docking confirmed that eight NSAIDs could stably bind to EPHB4. Crucially, we found that indomethacin exhibited a unique "dual-regulatory" pattern: it simultaneously targeted the protective target PTGER4 and the risk-associated target EPHB4. Molecular docking further substantiated, at the atomic level, that indomethacin possesses strong binding affinity for both targets, providing a structural biology explanation for its observed genetic effects.

Interpretation: This study provides, for the first time, robust causal evidence from both genetic and structural biology perspectives for the "double-edged sword" attribute of NSAIDs in EMS, and proposes a new paradigm of "target-oriented heterogeneous effects." We found that specific NSAIDs might inadvertently promote disease progression by activating the EPHB4 pathway, while indomethacin stands out as a key exception due to its unique dual-action mechanism. These findings not only offer a critical explanation for the current clinical controversy but also lay a solid scientific foundation for advancing EMS management from "empirical medication" towards "precision-based selection guided by molecular mechanisms."

Background: More than 500 patients have received cryopreserved ovarian tissue transplantation (OTT) worldwide, resulting in over 200 live births. Although guidelines recognize OTT as an effective fertility preservation method, there is no consensus on the optimal grafting site of OTT. This systematic review and meta-analysis aim to assess whether reproductive outcomes of OTT vary across different grafting sites.

Methods: A literature search was conducted through March, 2024, in PubMed, Embase and the Cochrane library, using the following terms: 'patient', 'fertility preservation', 'ovarian tissue transplantation' and 'live birth'. Studies including 5 or more subjects were included. Two reviewers independently selected the studies, collected the data and assessed the risk of bias. Heterogeneity and publication bias were evaluated using the I² statistic and Egger's test, respectively. A fixed-effect meta-analysis was performed only if I² was 0%, otherwise a random-effect model was applied. The primary outcome was the live birth rate (LBR). The secondary outcomes included the proportion of women who became pregnant, those who had at least one live birth, and those who underwent repeat OTT.

Results: Eighteen studies including 560 women were included. The estimated LBRs after transplantation to the remaining ovary and the pelvic peritoneum were 64% (20%-130%, I² = 74%) and 31% (15%-51%, I² = 21%), respectively. The LBRs after orthotopic transplantation (defined as transplantation to the remaining ovary, the pelvic peritoneum or both sites, which allowed spontaneous pregnancy), heterotopic transplantation and combined (orthotopic + heterotopic) transplantation were 44% (25%-69%, I² = 79%), 5% (0%-21%, I² = 0%) and 23% (4%-53%, I² = 19%), respectively. No publication bias was observed.

Conclusion: Reproductive outcomes of OTT vary across different grafting sites. The LBR of orthotopic transplantation is higher than that of heterotopic transplantation. Additionally, the LBR of OTT to the remaining ovary is higher than that to the pelvic peritoneum. The remaining ovary may therefore be a more suitable site for women undergoing OTT to preserve future fertility.

Registration number: CRD42023447618.

Non-obstructive azoospermia (NOA) is one of the most severe manifestations of male infertility, accounting for up to 70% of azoospermic cases and affecting approximately 1% of the male population. Advances in genomics and epigenetics have transformed our understanding of NOA from a primarily idiopathic condition into a biologically heterogeneous disorder driven by diverse molecular mechanisms. This review synthesizes the current knowledge of the genetic and epigenetic landscape of NOA, integrating chromosomal abnormalities, single-gene mutations, and non-coding RNA (ncRNA) dysregulation. First, we systematically examine classical and emerging chromosomal defects-including karyotype anomalies, Y-chromosome microdeletions, and structural rearrangements-that disrupt meiotic pairing and chromatin organization. Next, we explore syndromic and non-syndromic monogenic mutations affecting meiotic regulators, DNA repair factors, transcription regulators, and chromatin remodelers. Particular emphasis is placed on recently identified genes such as SYCP1, SYCE1 and HORMAD1, whose pathogenic variants are frequently linked to spermatogenic arrest. We then discuss the expanding role of ncRNAs-including microRNAs, PIWI-interacting RNAs, long non-coding RNAs, and circular RNAs-in regulating germ cell apoptosis, transposon silencing, and epigenetic reprogramming. Furthermore, we highlight the translational potential of these molecular insights (including gene variants, ncRNAs and protein) in clinical applications. Genotype-guided sperm retrieval, non-invasive biomarkers, and multi-omic approaches are discussed as promising tools to improve diagnosis and treatment. Moreover, we summarize current and emerging strategies for the treatment and fertility preservation of NOA. Finally, we identify persisting challenges, such as genotypic heterogeneity and incomplete functional validation, and emphasize the need to elucidate interactions between ncRNA and classical genetic pathways to uncover regulatory hierarchies underlying NOA. By integrating molecular genetics with testicular histopathology and clinical phenotypes, this review highlights emerging genetic and ncRNA biomarkers and underscores their potential applications in the clinical management of NOA. Ultimately, a comprehensive understanding of the genetic and epigenetic underpinnings of NOA will be essential for advancing precision diagnostics and improving reproductive outcomes in affected men.

Background: The posttranslational histone modification H3K9me3 is crucial for constitutive heterochromatin (cHC) and supports genome stability and gene regulation during development. This epigenetic mark persists in human sperm post histone-to-protamine transition and is transmitted to the embryo. Although H3K9me3 variability is linked to abnormal sperm parameters, its role in fertilization and embryo development remains unclear. Given its retention in sperm, aberrant H3K9me3 levels may underlie cases of unexplained male infertility.

Objective: Investigate the variability of H3K9me3 levels in sperm from normozoospermic men and assess its association with early embryo development and IVF outcomes.

Material and methods: H3K9me3 and histone H3 levels were quantified by Western blot in surplus sperm from 99 normozoospermic men undergoing IVF-treatment. Patients were stratified into quartiles based on the H3K9me3/H3 ratio. Pre-implantation embryo development was assessed by time-lapse imaging, focusing on nuclear precursor body (NPB) dynamics and morphokinetics. IVF outcomes were reported as cumulative biochemical and ongoing pregnancy rates per ovum pick-up and compared across H3K9me3/H3 quartiles.

Results: H3K9me3/H3 ratios exhibited substantial inter-individual variability among normozoospermic patients. Embryos from the third H3K9me3/H3 ratio quartile (Q3) demonstrated the highest proportion of zygotes with NPB clustering and faster, more consistent development through the first two cleavage divisions compared to other quartiles. A significant non-linear association was found between H3K9me3/H3 ratio and cumulative biochemical pregnancy rates: couples in the lowest quartile (Q1) had significantly reduced odds of biochemical pregnancy compared to Q3 (adjusted OR [95% CI]: 0.30 [0.09-0.97], p = 0.045). No significant association was found for ongoing pregnancy rates.

Discussion and conclusions: This study reveals that sperm H3K9me3 levels vary among normozoospermic men and correlate with early embryo development and biochemical pregnancy rates following IVF. However, no significant association was found with ongoing pregnancy, suggesting that additional mechanisms may determine long-term pregnancy viability. The non-linear relationship between H3K9me3/H3 ratio and embryo development suggests an optimal range for this epigenetic mark. These findings highlight the potential influence of paternal epigenetic variation, undetectable by standard semen analysis, on embryo quality and IVF outcomes. Further studies in larger cohorts are warranted to confirm these findings and clarify underlying mechanisms.

Background: Microdissection testicular sperm extraction (micro-TESE) is an effective method to retrieve sperm from non-obstructive azoospermia (NOA) patients. However, the predictive factors for sperm retrieval rate (SRR) remain confused. The goal of our study was to identify the role of testicular pathological morphometric parameters, including diameter of tubule (DT), height of spermatogenic epithelium (HSE), and thickness of basement-membrane (TBM) in NOA patients, and to develop a predictive model and nomogram to predict SRR based on these morphometric parameters.

Methods: This study involved two cohorts including 406 men with NOA. A retrospective cohort of 313 males with NOA who underwent micro-TESE at Northwest Women's and Children's Hospital (Xi'an, China) were included to build a prediction model of SRR. Then, another retrospective cohort of 93 males with NOA from Ren Ji Hospital (Shanghai, China) were recruited to validate the prediction model. The measurement of testicular morphometric parameters as well as the assessment of Johnsen score and pathological diagnostic types were performed by at least two pathologists. Testicular volumes as well as level of serum hormones including follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone (T) were also measured. Logistic regressions were used to test potential predictors of SRR. Area under curve (AUC) estimates was used to evaluate the predictive accuracy. The validation datasets were used to validate the prediction model by prediction accuracy.

Results: Our study demonstrated that DT and HSE were significantly longer in successful sperm retrieval group than in failed sperm retrieval group. In addition, DT and HSE were positively correlated with Johnsen score, testicular volume, and serum T, while, were negatively correlated with serum FSH and serum LH. On the contrary, TBM demonstrated exact opposite results. Moreover, univariate logistic analyses illustrated that longer DT and HSE was associated with a high SRR, respectively. Further multivariate logistic analyses constructed multi-variables models with better predictive abilities compared with single-variables models. A multi-variables model (predicting score = -0.612-0.018 × DT + 0.040 × HSE + 0.097 × Johnsen score-0.004 × serum FSH) was finally constructed with the best predictive ability (AUC = 0.839, sensitivity = 71.4% specificity = 77.5%, cut-off value = 0.489). A higher predicting score indicated a higher possibility of successful sperm retrieval. The predictive accuracy was 89.25% in the external validation dataset.

Conclusion: We report for the first time that DT and HSE have pretty ability to predict SRR in NOA patients.

Follitropin delta, a recombinant human follicle-stimulating hormone (r-hFSH), is administered using an individualized dosing algorithm based on serum anti-Müllerian hormone (AMH) levels and body weight and offers a different pharmacokinetic (PK) and pharmacodynamic (PD) profile compared to conventional gonadotropins. The retrospective analysis by Eggersmann et al. aimed to compare the cumulative live birth rate (CLBR) of r-hFSH delta versus r-hFSH alfa/beta using data from the German In-vitro Fertilization (IVF) registry (D.I.R.^®; Deutsches IVF-Register), encompassing 113,936 stimulation cycles from women aged 24-45 years between 2017 and 2022. The authors found no statistical differences in oocyte yield, pregnancy rate (PR), and live birth rate (LBR), and an increase in cumulative live birth rate (CLBR) in the group stimulated with r-hFSH delta. The study presents several methodological challenges, such as differences in dose exposure, insufficient adjustment for confounding variables, and the inclusion of diverse patient groups, which may affect the clarity of the comparative effectiveness of the treatments. Additionally, the exclusion of patients who did not undergo frozen embryo transfer may limit the interpretability and generalizability of the findings. While Eggersmann et al. added valuable and meaningful insights into the real-world effectiveness of r-hFSH delta, its conclusions warrant thoughtful consideration. Undertaking a comprehensive methodological assessment strengthens causal inferences drawn from observational data, especially when intended to inform clinical decision-making. This real-world study serves as an important piece of the broader evidence base, contributing to our understanding of assisted reproductive technology (ART) outcomes.