Ge Song

Non-obstructive azoospermia (NOA) is a severe male infertility disorder. This study aims to identify immune-related biomarkers for explore immune-related molecular signatures associated with NOA. Here, we integrated transcriptomic data from four independent cohorts (GSE145467, GSE45885, GSE45887, GSE9210) using an integrative pipeline combining differential expression analysis, weighted gene co-expression network analysis (WGCNA), and consensus-based multi-machine learning to identify an immune-related gene signature associated with NOA. We identified 16 immune-related differentially expressed genes through WGCNA, which were refined into a 12-gene panel— Hub NOA.Sig ( CD14, CD19, CMTM2, CXCL16, HSPA1L, HSPA2, IGF1, IL13, POMC, RBP1, STAB2, TYROBP )—using five feature selection algorithms (Bagged Trees, Bayesian, Boruta, LASSO, LVQ). A Random Forest classifier based on Hub NOA.Sig yielded good classification performance with an AUC of 0.87. This signature offers a transcriptomic reference for characterizing NOA heterogeneity and exploring immune-related molecular features. Single-sample gene set enrichment analysis (ssGSEA) revealed significant alterations in six immune cell populations—including effector memory CD4 ⁺ /CD8 ⁺ T cells, regulatory T cells, and plasmacytoid dendritic cells—and demonstrated strong correlations between hub genes and specific immune infiltrates. Furthermore, drug-gene interaction screening and molecular docking predicted loperamide- POMC , acitretin- RBP1 , and retinol- RBP1 as candidate drug-gene pairs for subsequent experimental validation. This work provides transcriptomic clues and candidate molecular targets for future mechanistic investigations into NOA-related male infertility.

Background: Recurrent spontaneous abortion (RSA) significantly impacts women’s health, yet the underlying biological mechanisms remain poorly defined. Understanding the molecular contributors to RSA is crucial for developing targeted interventions.Objective: This study aims to investigate the causal relationships between plasma proteins and RSA, focusing on the identification of potential therapeutic targets through multi-omic approaches.Methods: We utilized two-sample Mendelian randomization (MR) analyses integrating genome-wide association study (GWAS) data for both plasma proteins and RSA. Proteomic data were sourced from the UK Biobank-Plasma Proteome Project and deCODE Health Study. We further validated our findings through both bulk and single-cell RNA sequencing of clinical specimens, alongside quantitative real-time polymerase chain reaction and immunohistochemistry. A phenome-wide association study was also conducted to assess the safety and broader implications of identified targets.Results: Our analyses identified the interleukin 6 receptor (IL6R) as a key candidate, with elevated plasma levels correlating with increased RSA risk. Furthermore, IL6R was found to be upregulated in RSA-related endometrial and decidual tissues. The phenome-wide association study provided insights into potential side effects and additional therapeutic indications for IL6R.Conclusion: IL6R upregulation is mechanistically implicated in the pathogenesis of RSA, establishing it as a validated causal biomarker and a potentially actionable therapeutic target. This study not only highlights the role of IL6R in RSA but also supports its development into a therapeutic strategy with a comprehensive safety profile.

Background: PATL2 deficiency is a significant cause of female infertility. Although multiple PATL2 missense variants have been reported in prior studies, a number of these variants remain classified as variants of uncertain significance (VUS).Methods: We present a patient of primary infertility characterized by oocyte maturation disorders and fertilization failure. Comprehensive genetic analysis was conducted through whole-exome sequencing (WES) to identify pathogenic variants, followed by Sanger sequencing for familial co-segregation analysis. Reverse transcription (RT-PCR), cDNA sequencing and quantitative RT-PCR were performed to validate the effect of the variant on pre-mRNA splicing.Results: We identified compound heterozygous variants in the PATL2 gene by WES: a pathogenic splice-site splicing variant (c.223-14_223-2del) and a missense variant (c.877G>T) initially classified as a VUS. Sanger sequencing confirmed that the proband carried biallelic variants, whereas her sisters with either wild-type genotypes or a single heterozygous variant exhibited normal fertility, supporting the co-segregation of the identified variants. Critically, RNA assays demonstrated that the missense variant c.877G>T disrupts canonical splicing of PATL2, resulting in exon 12 skipping.Conclusion: This study provides the first experimental evidence that a PATL2 missense variant (c.877G>T) can exert its pathogenicity through aberrant splicing, supporting its pathogenic reclassification and elucidating a genotype-phenotype correlation for PATL2 missense variants through functional assays.

Ovarian endometrioma (OE), also known as “chocolate cysts,” is a cystic mass that develops in the ovaries due to endometriosis and is a common gynecological condition characterized by the growth of endometrial tissue outside the uterus, leading to symptoms such as dysmenorrhea, pelvic pain, and infertility. However, the precise molecular and cellular mechanisms driving this pathophysiology remain largely unknown, posing challenges for diagnosis and treatment. Here, we employed integrated single-cell transcriptomic profiling of over 52,000 individual cells from endometrial tissues of OE patients and healthy donors and identified twelve major cell populations. We identified notable alterations in cell type-specific proportions and molecular signatures associated with OE. Notably, the activation of IGFBP5⁺ macrophages with pro-inflammatory properties, NK cell exhaustion, and aberrant proliferation of IQCG⁺ and KLF2⁺ epithelium are key features and may be the potential mechanisms underlying the pathogenesis of OE. Collectively, our data contribute to a better understanding of OE at the single cell level and may pave the way for the development of novel therapeutic strategies.

Polycystic ovary syndrome (PCOS) is a reproductive endocrine disease, and results to opsomenorrhea or amenorrhea, hairy, acne, acanthosis, infertility, abortion. In the long term, PCOS may also increase the risk of endometrial cancer, diabetes, hypertension, dyslipidemia and other diseases. Till now there is no specific drug for PCOS due to the unclearness of the cause and pathogenesis, as current treatments for PCOS only target certain symptoms. Quercetin (QUR) is a flavonoid drug widely found in Chinese herbal medicines, fruits, leaves, vegetables, seeds and plants roots. Studies on other diseases have found that QUR has anti-oxidant, anti-inflammatory, anti-insulin resistance, anti-cancer and other effects. Some studies have shown that serum testosterone (T), luteinizing hormone (LH), the LH/follicule-stimulating hormone (FSH) ratio, fasting glucose, fasting insulin, HOMA-IR and lipid levels are reduced in PCOS patients with QUR treatment. However, the mechanisms of QUR in PCOS patients have not been completely elucidated. In this review, we retrospect the basic characteristics of QUR, and in vitro studies, animal experiments and clinical trials of QUR and plant extracts containing QUR in the treatment of PCOS. We also summarized the effects and mechanism of QUR in ovarian cells in vitro and PCOS model rats, the changes in relevant parameters after QUR administration in PCOS patients, and its potentially therapeutic applications.

Despite that gonadotropin-releasing hormone (GnRH) agonist pretreatment has been widely used before programmed frozen-thawed transfer (FET), its impact on live birth rates in ovulatory women remains uncertain. In the present study, we aim to determine if GnRH agonists pretreatment before FET improves live birth rates in women undergoing in vitro fertilization with FET. Programmed FET cycles conducted in four infertility centers were retrospectively collected and reviewed for eligibility from January 2016 and December 2017. Patient’s demographics, ovarian stimulation parameters, and pregnancy outcomes were compared between those given GnRH agonist pretreatment versus no pretreatment in ovulatory women undergoing FET cycles. A total of 6397 programmed cycles were screened for eligibility, of which 5049 cycles were included in the study for analysis. Compared with the group of no GnRH agonist pretreatment (n = 4143), women in the GnRH agonist group (n = 906) were older (33.0 vs 34.0, P < .001), had a higher proportion of subjects with previous transfer attempts and had a higher number of embryos transferred. After controlling for confounders, the logistic regression results showed that GnRH agonist pretreatment did not increase the odds of both clinical pregnancy (OR 0.92, 95% CI [0.70–1.20]), ongoing pregnancy (OR 0.91, 95% CI [0.69–1.19]) and live birth rates (OR 0.84, 95% CI [0.64–1.10]). However, when restricted to women who had no previous transfer attempts, women in the GnRH pretreatment group had lower odds of achieving live birth (OR 0.49, 95% CI [0.30–0.79]). Sensitivity analysis performed in patients with male factor infertility causes showed GnRH agonist pretreated group had lower live birth rates compared to no GnRH agonist pretreatment group (OR = 0.65, 95% CI [0.43–0.97]). Our findings suggested that GnRH agonist pretreatment does not bring additional benefits in live birth rate improvements for ovulatory women undergoing FET cycles. Therefore, the pros of using GnRH agonist to reduce premature ovulation should be weighed against the cons of prolonged time to pregnancy, discomforts resulting from pituitary suppression, and increased medical costs associated with GnRH agonist use.

This study is to analyze the effect of C-type natriuretic peptide (CNP) on sperm motility of asthenozoospermia and explore the influence mechanism of CNP on the reproductive system and sperm motility. Our results showed that the concentration of CNP in asthenospermia patients’ semen was lower than in normal people’s. The motility of sperm could be improved markedly by CNP and 8-Br-cGMP, while the effect of CNP was inhibited by NPR-B antagonist and KT5823. In the asthenozoospermia mouse model induced by CTX, CNP injection could improve sperm motility in the epididymis, alleviate tissue damage in the testes and epididymis, and increase testosterone levels. The asthenospermia mouse model showed high activity of MDA and proinflammatory factors (TNF-α, IL-6), as well as low expression of antioxidants (SOD, GSH-Px, CAT) in the testis and epididymis, but this situation could be significantly ameliorated after being treated with CNP. Those studies indicated that the concentration of CNP in the semen of asthenospermia patients is lower than in normal people and could significantly promote sperm motility through the NPR-B/cGMP pathway. In the asthenospermia mouse model induced by CTX, CNP can alleviate the damage of cyclophosphamide to the reproductive system and sperm motility. The mechanism may involve increasing testosterone and reducing ROS and proinflammatory factors to damage the tissue and sperm.

This study aimed to evaluate the clinical characteristics, pregnancy outcomes and prognostic factors for pregnancy of female with chromosomal abnormalities (CAs) after artificial insemination with donor's sperm (AID) treatment. A retrospective case–control study was analyzed by using the data of 29 female patients with CA and 116 controlled patients with normal karyotype (1:4 ratio) who underwent AID cycles at Guangdong Family Planning Special Hospital from January 2011 to December 2017. In all cases, reproductive histories were collected, and the cytogenetic analysis was performed by Trypsin-Giemsa banding and karyotyping. The embryos were fertilized via intracervical or intrauterine insemination. Clinical characteristic variables were compared. The prevalence of CA was found to be 0.29% in the whole AID population. The live birth rates of CA group and controlled group were 41.4% and 31.0% (P = .29) respectively. Compared to normal karyotype group, patients with CA showed higher rate of primary infertility (93.1% vs 75.9%, P = .049); Multivariate analysis demonstrated that ovarian stimulation (odds ratio, 3.055; 95% confidence interval, 1.421–6.568; P = .004) was associated with adverse pregnancy outcomes in female patients with AID treatment. For the infertility CA patients who were phenotypically normal, AID was a suitable choice, whereas ovarian stimulation results in an improvement in the pregnancy rate.

Mammalian ovaries consist of follicles as basic functional units. Each follicle comprised an innermost oocyte and several surrounding flattened granulosa cells. Unlike males, according to the initial size of the primordial follicle pool and the rate of its activation and depletion, a female’s reproductive life has been determined early in life. Primordial follicles, once activated, will get into an irreversible process of development. Most follicles undergo atretic degeneration, and only a few of them could mature and ovulate. Although there are a lot of researches contributing to exploring the activation of primordial follicles, little is known about its underlying mechanisms. Thus, in this review, we collected the latest papers and summarized the signaling pathways as well as some factors involved in the activation of primordial follicles, hoping to lead to a more profound understanding of the cellular and molecular mechanisms of primordial follicle activation.