Polycystic Ovary Syndrome (PCOS) is the most common endocrine disorder in reproductive-age women. Heat shock 70 kDa protein 1B (HSPA1B) is critical for stress response and inflammation, but its role in PCOS remains unknown. To validate HSPA1B as a key pathogenic gene in PCOS and investigate its regulatory mechanisms. Two PCOS datasets (GSE155489, granulosa cells; GSE54250, whole blood) were analyzed for differentially expressed genes. HSPA1B diagnostic value was assessed by ROC curve. Dihydrotestosterone (DHT)-induced KGN cell and dehydroepiandrosterone (DHEA)-induced mouse PCOS models were established. siRNA/adenoviral silencing, overexpression, methylated RNA immunoprecipitation (MeRIP), RNA immunoprecipitation (RIP), chromatin immunoprecipitation (ChIP), luciferase assay, actinomycin D chase, MTT, flow cytometry, ELISA, and Western blot were used to evaluate HSPA1B effects on granulosa cell function and dissect the YY1-NSUN7-m5C-HSPA1B axis. HSPA1B was identified as a core DEG, upregulated in patient blood and DHT-induced KGN cells (AUC = 0.9273). Silencing HSPA1B reversed DHT-induced injury: restored viability, reduced IL-6/IL-1β, decreased apoptosis, stabilized mitochondrial potential, reduced ROS, and inhibited M1 macrophage polarization (all P < 0.05). In vivo, HSPA1B silencing ameliorated ovarian pathology, corrected sex hormones (restored estradiol [E2], decreased testosterone [T]), and alleviated inflammation (all P < 0.01). Mechanistically, NSUN7 bound HSPA1B mRNA 3' UTR, enhancing stability via m5C methylation; Yin Yang 1 (YY1) activated NOP2/Sun RNA methyltransferase 7 (NSUN7) transcription by promoter binding. Rescue experiments established the YY1-NSUN7-m5C-HSPA1B axis. This study revealed the YY1-NSUN7- m5C -HSPA1B epigenetic axis in PCOS pathogenesis, providing a foundation for HSPA1B-targeted therapy.

Unexplained recurrent pregnancy loss affects many fertile couples who want to conceive. Possibly, the maternal immune system is involved in the underlying mechanism of such unexplained cases and immune-suppressing corticosteroids can be used to modulate this system and improve pregnancy outcome. As a high quality, sufficiently powered randomized controlled trials in the unselected unexplained RPL population is currently lacking, the PREMI (Prednisolone administration in patients with unexplained REcurrent Miscarriages) trial was designed. Here, we discuss the rationale for the administration of prednisolone, including its potential mechanisms, safety, administration protocol and study population.

Ovarian cancer is one of the most lethal gynecological malignancies with poor prognosis due to high drug resistance and low tumor susceptibility. This study investigated the cytotoxic effects of four sterols of fungal origin, ergosterol (ES), cholesterol (CS), β-sitosterol (βS), and lanosterol (LS) on human ovarian cancer SKOV3 cells, revealing that they exert their effects by correlating with autophagy and ferroptosis‑related signaling, and analyzed the structure-activity relationship. SKOV3 tumor cells were treated with four types of sterols for 24 h. Cell viability, cell cycle, apoptosis, mitochondrial membrane potential, and the expression of proteins associated with autophagy and ferroptosis were assessed, and the correlation between their structure and antitumor activity was analyzed. All four sterols inhibited SKOV3 cell proliferation in a dose-dependent manner, with the IC50 values ranked as LS (28.32 μg/mL) < CS (32.07 μg/mL) < βS (73.53 μg/mL) < ES (78.67 μg/mL). The four sterols significantly suppressed cell migration, induced G0/G1 phase arrest and apoptosis, reduced mitochondrial membrane potential, and caused mitochondrial structural damage. Meanwhile, they markedly up-regulated the expression of LC3II/LC3I and P53, and down-regulated the expression of SLC7A11 and GPX4. The number of conjugated double bonds in sterols was negatively correlated with IC50, cell viability and migration rate, and positively correlated with apoptosis rate and key protein expression, the substituents at C24, C28 and C29 of the side chain also enhanced the cytotoxicity. These findings indicate that fungal sterols are associated with correlated regulation of the autophagy-ferroptosis network and P53 activation in SKOV3 cells, providing a theoretical basis for anti-ovarian cancer drug development.

Antisperm antibodies (ASA) have been implicated in reproductive failure in cows. Present study investigated 58 crossbred cows, of which 38 (normal-cows, NC, n = 16; repeat-breeder-cows, RBC, n = 22) were used for ASA analyses and fertility evaluation, while subsequent bacteriological component included additional 20 cows. ASA levels were assessed from maiden to conceptional-estrus using mixed-antiglobulin reaction (MAR-IgG and MAR-IgA) tests and ELISA, and candidate spermatozoal-bacterial protein pairs were assessed for sequence and three-dimensional structural similarity. MAR-IgG and MAR-IgA levels were significantly lower during conceptional-estrus than during maiden-estrus in both NC and RBC cows, whereas differences between NC and RBC at same estrus stage were less pronounced, indicating that ASA levels were closely associated with conception status than with fertility classification. Receiver operating characteristic analysis based on cow-level measurements showed that MAR-IgG and MAR-IgA discriminated conceptional from non-conceptional status, with area under the curve values of 0.799 and 0.806, respectively. Head-bound spermatozoa represented predominant antibody binding, suggesting that sperm head antigens constitute major ASA targets. Bacillus spp., Escherichia coli and Staphylococcus spp. were predominant bacterial isolates recovered from the genital tract. Sequence and structural analyses of previously identified cross-reactive sperm and bacterial proteins revealed substantial peptide-level similarity and structural-overlap, and structural-superimposition produced low root mean square deviation values, indicating close structural similarity between sperm and bacterial proteins. Collectively, these findings indicate that lower ASA levels are associated with successful conception and support molecular mimicry between genital bacterial and sperm proteins as a plausible mechanism contributing to reproductive immunoinfertility in cows.

Immunocontraceptive vaccines have emerged as a promising, non-invasive strategy for regulating fertility in wildlife, domestic animals, and humans, particularly in species such as elephants where conventional population control methods are often impractical or ethically challenging. These vaccines act by eliciting immune responses against critical reproductive targets, thereby disrupting gametogenesis, fertilization, or endocrine regulation. The major categories include gonadotropin-releasing hormone (GnRH)-based vaccines, zona pellucida (ZP)-based vaccines, and gamete-specific antigen vaccines. Anti-GnRH vaccines induce neutralizing antibodies that suppress the hypothalamic-pituitary-gonadal axis by inhibiting luteinizing hormone and follicle-stimulating hormone secretion, ultimately blocking steroidogenesis and gamete production. In contrast, ZP-based vaccines target oocyte surface glycoproteins, preventing sperm binding and fertilization. Emerging molecular targets, including sperm-egg interaction proteins such as IZUMO1 and JUNO, offer enhanced specificity. These vaccines predominantly stimulate humoral immune responses, with antigen-specific antibody production playing a central role, although cell-mediated immunity may contribute to sustained effects. In elephants, vaccines such as porcine zona pellucida (PZP) and GnRH-based formulations (e.g., GonaCon) have shown variable success in inducing reversible infertility, though challenges remain regarding booster requirements, duration of efficacy, and potential side effects. Advances in scientific technologies, particularly peptide engineering and viral-vector platforms, have contributed significantly to the development of immunocontraceptive vaccines by enhancing antigen specificity, immunogenicity, and overall vaccine performance. However, variability in immune responses, species-specific efficacy, and ethical considerations continue to limit widespread application. This review aims to help understanding the molecular and immunological mechanisms are essential for optimizing safe, effective, and sustainable fertility control strategies.

Uterine fibroids are benign tumours of the myometrium. For large and/or multiple fibroids, there are no fertility-sparing pharmacological therapies. Better understanding of fibroid pathophysiology is key to developing such treatments. Current understanding of the pathophysiology of fibroid development has largely centred on hormone dependent growth and regression. The uterus is highly immunologically active, and pathological lesions from other benign uterine conditions exhibit local differences in immune cell populations compared to neighbouring analogous tissue. Characterising the differences in the local immune cells between fibroid tissue and non-diseased myometrium will inform understanding of fibroid pathophysiology and future therapeutic targets. This systematic review synthesises current evidence on the immune cell presence within uterine fibroids. Existing studies suggest a trend towards increased macrophage infiltration in fibroid tissue compared to matched myometrium, and reduced macrophage presence following hormonal therapy. There are conflicting reports on fibroid leucocyte and mast cell populations. Only a limited number of studies investigated a broader panel of immune cells, including dendritic, natural killer and T cells. Considerable methodological heterogeneity was observed across studies, alongside a frequent lack of demographic reporting, and variable use of matched fibroid-adjacent and healthy myometrial controls. To better characterise the immunological environment of uterine fibroids and the hormonal mechanisms that may govern it, future studies should employ standardised methodology, comprehensive immune cell panels and larger sample sizes that are segregated by menstrual cycle phase.

Gestational diabetes mellitus (GDM) adversely affects maternal and offspring health, and the early-life microbiome is implicated in developmental programming. This systematic review quantitatively evaluated the association between maternal GDM and alterations in the gut and oral microbiomes of offspring. We systematically searched PubMed, Web of Science, and Embase (up to August 2025) for eligible studies adopting high-throughput sequencing techniques. Random-effects meta-analysis was performed for alpha diversity metrics, while findings on beta diversity and microbial taxonomy were summarized narratively. Twenty-one studies were included. The meta-analysis revealed no statistically significant differences in gut microbial alpha diversity between infants exposed to maternal GDM and unexposed controls. Beta diversity exhibited substantial inter‑study heterogeneity, with no consistent shifts observed. In infants born to mothers with GDM, several microbial taxa including Gammaproteobacteria, Enterobacteriaceae, Micrococcaceae, Propionibacteriaceae, Rikenellaceae, Megasphaera, Staphylococcus, Collinsella, Rothia, Pelomonas, Veillonella, Shewanellaceae and Prevotella tended to decrease in abundance, whereas Escherichia tended to increase. Notably, no microbial taxon presented consistent abundance changes in three or more studies, and these repeated findings were based on limited evidence. Delivery mode appeared to have minimal influence on offspring gut alpha diversity. Collectively, maternal GDM may not be strongly linked to shifts in offspring gut or oral microbial alpha diversity. However, the consistent dysregulation of specific microbial taxa in GDM-exposed offspring may exert potential impacts on offspring immune and metabolic development. Further longitudinal multicenter investigations are warranted to elucidate the causal relationships and long-term health implications of GDM-induced microbial alterations in children.

Endometriosis is associated with immune dysregulation yet circulating cytokine measurements have shown limited discriminatory value. We investigated whether peripheral blood mononuclear cells (PBMCs) from women with endometriosis exhibit altered basal or phytohemagglutinin (PHA)-induced cytokine secretion. In this single-center case-control study, PBMCs from 36 women with laparoscopically and histopathologically confirmed endometriosis and 44 laparoscopy-confirmed controls without endometriosis were cultured for 24 h under unstimulated and PHA-stimulated conditions. Supernatant concentrations of 27 cytokines, chemokines and growth factors were measured using a Bio-Plex/Luminex assay. Cytokine concentrations were analyzed using linear mixed-effects models including group, condition, and group × condition interaction terms, with false-discovery-rate correction. PHA induced broad cytokine responses, with significant condition effects for 20 of 27 analytes in the full panel and 14 of 15 analytes in the robust quantifiable panel. A significant group × condition interaction was observed for IL-5; however, this result was based on very sparse PHA data and was not retained in a robust restricted panel of adequately quantified analytes. Exploratory multivariate analyses did not show clear group separation. Overall, these findings do not support the presence of a broad and reproducible endometriosis-associated cytokine-response profile in 24-h bulk-PBMC cultures stimulated with PHA-L at 5 µg/mL. Within this experimental framework, any between-group differences appeared subtle, analyte-specific, and sensitive to data completeness. These results are compatible with, but do not directly demonstrate, the concept that disease-related immune alterations may be more prominent in specific immune-cell subsets or local pelvic compartments than in non-specifically stimulated circulating PBMC cultures.

During follicular development, granulosa cell autophagy is dynamically involved in follicular growth, atresia, estrogen and progesterone synthesis, luteinization, and luteal regression. Granulosa cell autophagy is regulated by gonadotropins, estrogen, progesterone, oxidative stress, and multiple signaling pathways. Conversely, granulosa cell autophagy can also influence estrogen and progesterone synthesis by regulating lipid metabolism, mitochondrial quality control, and granulosa cell viability.However, the biological effects of autophagy are highly context-dependent. Moderate autophagy is an important mechanism for maintaining granulosa cell homeostasis and promoting follicular development, whereas excessive autophagy or impaired autophagic flux may lead to granulosa cell dysfunction, increased apoptosis, follicular atresia, and reproductive endocrine disorders. Dysregulated granulosa cell autophagy has been closely associated with ovulatory disorders such as polycystic ovary syndrome and premature ovarian insufficiency. Therefore, promoting follicular development through the regulation of autophagy may provide an important therapeutic entry point for ovulatory disorders. This review summarizes the roles and underlying mechanisms of granulosa cell autophagy in follicular development, follicular atresia, hormonal regulation, and ovulatory disorders, and proposes that maintaining autophagic homeostasis may represent a potential strategy for promoting follicular development and improving female reproductive health.

Preeclampsia (PE) is a pregnancy-specific syndrome characterized by an intense systemic inflammatory response and monocyte activation, with increased production of pro-inflammatory cytokines, including tumor necrosis factor-α (TNF-α). Evaluation of TNF-α receptor expression (TNFR1 and TNFR2) in monocyte subpopulations may provide new insights into the inflammatory mechanisms underlying this syndrome. In this study, monocytes from 20 women with PE and 20 normotensive (NT) pregnant women were analyzed by flow cytometry for M1-like (TLR4, CD64) and M2-like (CD163, CD206) markers, as well as TNFR1 and TNFR2 expression. Plasma levels of TNF-α, interleukin (IL)-1β, transforming growth factor beta (TGF-β), IL-10; and the soluble forms of TNF-α receptors, sTNFR1 and sTNFR2, were measured by ELISA. Monocytes from women with PE exhibited increased expression of TLR4 and CD64 and reduced expression of CD163 and CD206 compared with the NT group. In addition, a higher percentage of both M1-like and M2-like monocytes expressing TNFR1 and TNFR2 were observed in PE. Plasma concentrations of TNF-α, IL-1β, TGF-β, and sTNFR1 were significantly increased, whereas IL-10 levels were reduced in women with PE. No significant differences were observed in sTNFR2 levels between groups. These findings indicate that PE is associated with a predominance of M1-like monocytes and a reduction in the M2-like phenotype, contributing to the systemic inflammatory state characteristic of the syndrome. This imbalance appears to be linked to enhanced activation of the TNF-α/TNFR1 axis and dysregulated TNFR2-mediated regulatory signaling, highlighting TNF-α-driven immune dysregulation as a potential contributor to PE pathophysiology.

Autoimmune-associated reproductive dysfunction is frequently linked to ovarian immune infiltration and circulating ovarian autoantibodies, yet it remains unclear whether circulating factors alone can initiate immune changes in the ovary before overt histopathology is detectable. In prior work using interferon-γ (IFN-γ) AU-rich element deletion (ARE) mice, chronic inflammation was associated with increased ovarian CD8[+] T cell infiltration and reproductive dysfunction. Here, we tested whether circulating factors from ARE mice are sufficient to induce early ovarian immune remodeling in healthy hosts. Pooled plasma from WT, ARE[+/-], or ARE[-/-] female donors was administered intraperitoneally to age-matched WT female recipients, and tissues were harvested on day 8. Ovaries were assessed by H&E histopathology and CD8 immunohistochemistry; spleen and draining lymph nodes (dLNs) were analyzed by flow cytometry. In parallel, circulating anti-ovarian antibody levels were quantified in donors, and genotype-dependent ovarian tissue binding was evaluated by incubating Rag1[-/-] ovarian sections with donor plasma followed by immunofluorescent detection and whole slide image quantification. Passive plasma transfer did not induce overt ovarian histopathology by H&E at day 8. However, CD8 immunohistochemistry revealed a genotype-dependent increase in ovarian CD8[+] cell accumulation, highest in ARE[-/-] plasma recipients. Moreover, flow cytometry showed increased CD8[+] T cell frequencies in dLNs in ARE[-/-] plasma recipients. Donor ARE[-/-] mice exhibited elevated circulating ovarian autoreactivity and their plasma showed increased binding to Rag1[-/-] ovarian tissue ex vivo. Together, these data indicate that short-term exposure to disease-associated plasma can initiate an ovarian immune shift.

Natural killer (NK) cells are key regulators of endometrial receptivity and placentation, and aberrant activation of peripheral blood NK (pNK) cells has been implicated in recurrent pregnancy loss (RPL) and repeated implantation failure (RIF). This study investigated adhesion molecule expression on pNK cells in women with primary RPL (pRPL), secondary RPL (sRPL), and unexplained infertility (uINF), and its associations with NK-cell phenotypes, cytotoxicity (NKC), and T-helper (Th)-1/Th2 (Th1/Th2) immune balance. Expression of adhesion markers LFA-1, VLA-1, VLA-2, and ICAM-1 on NK cells was assessed by flow cytometry. NKC was measured using the K562 assay, and Th1/Th2 cytokine ratios were determined by intracellular cytokine staining using flow cytometry. Total CD56[+] NK cell proportions and overall NKC did not differ among groups. Expression of LFA-1, VLA-1, VLA-2, and ICAM-1 on total NK cells was also comparable. However, Th1/Th2 cytokine ratios were significantly lower in the sRPL group. Stratification by NKC revealed that women with RPL and higher NKC exhibited significantly increased ICAM-1 expression on cytotoxic subsets (CD56[+]CD16[+] and CD56[+]CD57[+]). NKC positively correlated with ICAM-1-expressing NK subsets, while Th1/Th2 ratios were associated with mature and activated NK phenotypes in women with RPL. Although overall adhesion molecule expression did not differ by clinical classification, elevated ICAM-1 expression was associated with increased NKC and pro-inflammatory immune signatures in RPL but not in uINF, suggesting an association between ICAM-1 expression on NK cell subsets, increased NKC, and altered Th1/Th2 responses in RPL.

Endometritis, which can cause endometrial fibrosis, is one of the key factors leading to the decline of pregnancy and reproduction rates in cows. Macrophages can be polarized into M1 type pro-inflammatory phenotype to aggravate endometrial injury under pathological conditions. Paeoniflorin (PF) is a single glucoside extracted from Paeoniaceae, which has anti-fibrosis and anti-inflammatory effects. However, its effect on fibrosis in endometritis in cows is unknown. In this study, we examined uterine fibrosis and macrophage polarization in endometritic cows, and established a macrophage-bovine endometrial stromal cell (BESC) co-culture system to clarify PF's protective mechanism. In vivo, endometritic cattle showed prominent collagen accumulation, elevated M1 markers CD86/iNOS, reduced epithelial E-cadherin, and increased fibrotic markers N-cadherin, α-SMA and Vimentin. In vitro, LPS-induced M1 macrophages were confirmed as a vital driver of BESC fibrosis, while M0 unpolarized or IL-4-stimulated M2 macrophages exerted no fibrotic effects. Mechanistically, PF suppressed NF-κB/STAT1 phosphorylation to block M1 polarization and reverse M1-mediated BESC fibrosis. In summary, the results of the present study indicate that PF attenuates endometrial fibrosis in cows by inhibiting M1 polarization of macrophages through the NF-κB and STAT1 pathway. These results provided insights into the exploration of therapeutic targets and intervention strategies for endometritis in cows.

Lactate is a key metabolite of glycolysis. Since the proposal of the lactate shuttle hypothesis, accumulating evidence indicates that lactate is not only an essential energy substrate but also a key signalling molecule that regulates cellular functions under both physiological and pathological conditions. Uterine spiral artery (SpA) remodelling is a critical event in successful pregnancy. It involves complex interactions among trophoblasts, vascular smooth muscle cells (VSMCs), decidual immune cells, and endothelial cells (ECs). Similar to the tumour microenvironment, physiological hypoxia at the maternal-foetal interface during early pregnancy drives decidual cells, blastocysts, trophoblasts, and immune cells to undergo aerobic glycolysis, leading to local lactate accumulation and the formation of an acidic microenvironment. This review systematically elucidates the role and molecular mechanisms of lactate metabolism in SpA remodelling. It explores the association between dysregulated lactate metabolism and recurrent pregnancy loss (RPL), preeclampsia (PE), and foetal growth restriction (FGR). The findings of this review indicate that targeting lactate metabolism may represent a promising strategy for the management of pregnancy complications. Further in-depth mechanistic research and clinical obstetric research are warranted.