PROBLEM:Endometriosis (EMS) is a gynecological condition that is associated with chronic pelvic pain and inflammation. Evidence supports the idea that natural killer (NK) cell dysfunction contributes to EMS pathogenesis; nevertheless, the role of T cell immunoreceptors with Ig and ITIM domains (TIGIT) and lymphocyte activation gene 3 (LAG3) in the development of endometriosis-associated immunological abnormalities is not yet reported. METHOD OF STUDY:The present study used multicolor flow cytometry to compare the frequency of NK cells expressing TIGIT and LAG3 in the peripheral blood (PB) and peritoneal fluid (PF) of women with and without endometriosis. The levels of soluble form of LAG3 (sLAG3) were compared between EMS and non-EMS participants in both PB and PF. RESULTS:The number of NK cells expressing TIGIT increased in the PF of EMS compared to the other benign gynecological conditions (OBGC) (Pv = 0.0089); however, this difference was not statistically significant in PB. In contrast, an increased frequency of LAG3+NK cells was detected in both PF and PB in EMS compared to the controls. Additionally, a higher level of sLAG3 was stated in both the PB and PF of EMS patients. CONCLUSIONS:An impaired number of TIGIT+ and LAG3+NK cells was detected in the peritoneal microenvironment of EMS, which may be responsible for impaired immune surveillance, promoting the survival of ectopic lesions, and contributing to the evolution of EMS.
OBJECTIVE:Repeated implantation failure (RIF) is often linked to immune dysregulation, particularly an elevated Th1/Th2 ratio that impairs embryo implantation. While several immunotherapies have been used, their efficacy and safety remain unclear. This study evaluated the clinical efficacy and immunomodulatory effects of tacrolimus treatment in women with RIF exhibiting elevated Th1/Th2 ratios. METHODS:This was a randomized, double‑blind, placebo‑controlled trial conducted between April 2024 to May 2025. A total of 153 RIF patients were screened, and 91 patients with a Th1/Th2 ratio ≥10.3 were identified. After exclusions, 80 patients were randomized 1:1 to tacrolimus or placebo. Patients were administered tacrolimus or placebo for 16 days, beginning 2 days before embryo transfer and continuing until the pregnancy test day. The primary outcome was the clinical pregnancy rate. Live birth was prespecified as the key secondary outcome. Additional secondary outcomes included miscarriage, ongoing pregnancy, multiple gestation, changes in the Th1/Th2 ratio, and adverse events. RESULTS:Clinical pregnancy occurred in 17 of 40 patients (42.5%) in the tacrolimus group compared with 6 of 40 patients (15.0%) in the placebo group (RR = 2.83, 95% CI: 1.30-6.44; p = 0.0125), corresponding to an absolute risk difference of 27.5% and a number needed to treat (NNT) of 4. Live birth occurred in 11 of 40 patients (27.5%) versus 3 of 40 patients (7.5%), respectively (RR = 3.67, 95% CI: 1.21-11.63; p = 0.0367), with an absolute risk difference of 20.0% and an NNT of 5. The Th1/Th2 ratio decreased significantly after treatment in the tacrolimus group (17.01 ± 3.99 to 12.56 ± 2.78, p < 0.0001), whereas no significant change was observed in the placebo group. No severe adverse events or treatment discontinuations occurred. CONCLUSION:Tacrolimus treatment improved clinical pregnancy outcomes and effectively reduced elevated Th1-dominant immune responses in RIF patients. These findings support the use of tacrolimus as an immunosuppressive therapy for RIF patients with elevated Th1/Th2 ratios. TRIAL REGISTRATION:This trial was registered with the Iranian Registry of Clinical Trials (IRCT20231127060207N1).
AIM:To evaluate the predictive value of first and second trimester hematological inflammatory indices-including the novel cumulative inflammatory index (IIC) and mean corpuscular volume-to-lymphocyte ratio (MCVL)-for the subsequent development of gestational diabetes mellitus (GDM). Additionally, the study will assess whether these markers differ between GDM subgroups managed with diet alone versus insulin therapy. METHODS:This case-control study enrolled 150 pregnant women, with 75 diagnosed with GDM and 75 serving as healthy controls. Hematological inflammatory indices-namely, NLR, MLR, PLR, dNLR, AISI, SII, SIRI, IIC, and MCVL-were calculated from first- and second-trimester complete blood count parameters. RESULTS:First-trimester inflammatory indices (NLR, MLR, PLR, dNLR, AISI, SII, SIRI, IIC, and MCVL) were significantly elevated in the GDM group compared to controls (all p < 0.05). NLR demonstrated the highest discriminative accuracy (AUC 0.782; 95% CI: 0.709-0.855), followed by SIRI (AUC 0.778) and IIC (AUC 0.775). Among GDM patients, 22 (29.3%) required insulin therapy. Insulin-treated patients exhibited significantly higher SIRI_1 than diet-managed patients (p = 0.025), and showed a substantially elevated inflammatory burden across multiple second-trimester indices. First-trimester fasting glucose was positively correlated with several second-trimester inflammatory indices, including AISI_1 (r = 0.351, p = 0.002) and SII_2 (r = 0.342, p = 0.003). CONCLUSION:First-trimester hematological inflammatory indices, including the novel IIC and MCVL, are associated with subsequent development of GDM. Furthermore, these indices offer moderate predictive accuracy when derived from routine antenatal blood tests. The clinical usefulness of these findings necessitates prospective validation in larger, multicenter cohorts.
PURPOSE:Sperm quality is a key contributing factor in infertility. This study aimed to investigate the causal relationship between gut microbiota genera and abnormal spermatozoa (defects in sperm count, motility, or morphology) and to evaluate potential mediating factors. PATIENTS AND METHODS:Summary statistics for gut microbiota genera, abnormal spermatozoa, microbiota metabolic pathways, immune cell traits, and serum bioavailable testosterone (BioT) levels were extracted from publicly available GWAS databases. A fixed-effects meta-analysis was performed on gut microbiota genera from the MiBioGen and DMP cohorts, followed by two-sample Mendelian randomization (MR) analysis to evaluate causal associations. To strengthen reliability, a triangulation framework incorporating MR analyses, HyPrColoc colocalization analysis, and Steiger directionality tests was employed. Mediation analysis focused on microbiota metabolic pathways, immune cell traits, and BioT levels as potential mediators. RESULTS:Eight gut microbiota genera showed significant causal associations with abnormal spermatozoa: four risk genera (Lachnospiraceae UCG001.id.11321, Coprococcus 1.id.11301, Barnesiella, Dorea) and four protective genera (Clostridium sensu stricto 1.id.1873, Ruminiclostridium 6.id.11356, Lachnospiraceae_noname, Veillonella). The triangulation framework validated these causal relationships. Although two genera affected BioT levels, formal mediation analysis did not confirm BioT, immune cells, or metabolic pathways as mediators, suggesting novel gut-testis axis mechanisms. CONCLUSION:This large-scale MR analysis establishes a causal connection between specific gut microbiota genera and abnormal spermatozoa. The "causation without colocalization" pattern and null mediation findings suggest distant regulatory mechanisms and novel gut-testis axes, offering microbiota-targeted therapeutic strategies for male infertility.
BACKGROUND:Endometriosis (EMs) is an estrogen-dependent chronic inflammatory disease, and its pathogenesis remains unclear. Although the placenta is an organ with tumor-like characteristics, its development, including its invasive function, is tightly controlled. One of the mechanisms is that extracellular vesicles (EVs) released by the placenta play an important role in this regulation. Placental extracellular vesicles carry functional proteins and regulatory RNAs. Our previous studies reported that placental extracellular vesicles inhibit the growth of ovarian cancer and cervical cancer both in vitro and in vivo. In recent years, ferroptosis, a novel form of cell death driven by iron-dependent lipid peroxidation, has been found to play a key role in the development of EMs. METHODS:Extracellular vesicles (EVs) derived from the placenta were isolated and identified, and their morphology and size distribution were characterized by transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA).In vitro experiments were conducted using endometriosis cells as a model. Cell viability was assessed using the CCK-8 assay. The expression of the ferroptosis-related molecules GPX4 and SLC7A11 was detected by Western blot and qRT-PCR, while lipid peroxidation and oxidative stress were evaluated by measuring malondialdehyde (MDA) and reactive oxygen species (ROS) levels. In addition, the ferroptosis inhibitor Ferrostatin-1 (Fer-1) was used for intervention experiments to verify the role of ferroptosis in this process. Potential miRNAs were further predicted using bioinformatic databases, and the regulatory mechanism was validated through miRNA transfection experiments, dual-luciferase reporter assays, and SLC7A11 overexpression rescue experiments. In vivo, a mouse model of endometriosis was established to evaluate the effect of EVs on the growth of ectopic lesions. RESULTS:The current study demonstrated that placental EVs delivered miRNA-26a-5p, which downregulated SLC7A11, led to increased lipid peroxidation and triggered ferroptosis, and ultimately inhibited the progression of endometriosis. Fer-1 effectively reversed these effects. In vivo, placental EVs significantly reduced ectopic lesion volume in mice. CONCLUSION:This study revealed that placental EVs induced ferroptosis through miRNA-26a-5p/SLC7A11-GPX4 axis, providing a potential therapeutic target for endometriosis.
PROBLEM:Preeclampsia (PE), characterized by hypertension and proteinuria, involves placental dysfunction linked to oxidative stress and ferroptosis. The role of estrogen receptor α (ERα) in hypoxia-induced trophoblast ferroptosis remains unclear. METHOD OF STUDY:A PE rat model was established using L-NAME injection. Placental tissues were analyzed for protein expression via Western blot. In vitro, HTR-8/SVneo cells were cultured under hypoxic conditions to simulate PE-related hypoxia. ERα was knocked down using siRNAs, and acyl-CoA synthetase long-chain family member 4 (ACSL4) was overexpressed via plasmid transfection. Cell viability, apoptosis, migration, oxidative stress parameters, and ferroptosis-related indicators were assessed using CCK-8, flow cytometry, Transwell assays, and specific biochemical kits. In addition, primary syncytiotrophoblast (STB) cells were isolated from placentas of healthy and PE rats for validation experiments. The transcriptional regulation of ACSL4 by ERα was examined using dual luciferase reporter assays. RESULTS:In PE rat placentas, ERα and ACSL4 were significantly upregulated, while GPX4 and SLC7A11 were downregulated. In addition, the lipid peroxidation products 4-HNE and MDA were significantly increased in PE placental tissues. Hypoxia induced ferroptosis in HTR-8/SVneo cells, characterized by increased Fe2 + levels, elevated 4-HNE and MDA levels, lipid peroxidation, and activation of the ERα/ACSL4 axis. Knockdown of ERα attenuated hypoxia-induced cell injury, oxidative stress, and ferroptosis, whereas ACSL4 overexpression reversed these protective effects. Furthermore, experiments in primary STB cells isolated from PE rats showed that ERα knockdown similarly alleviated ferroptosis-associated injury and suppressed ACSL4 expression. Mechanistically, ERα bound directly to the ACSL4 promoter and transcriptionally activated its expression. CONCLUSIONS:The ERα-ACSL4 axis plays a critical role in hypoxia-induced ferroptosis in placental trophoblasts. ERα transcriptionally activates ACSL4, promoting lipid peroxidation and ferroptosis, thereby contributing to placental dysfunction in PE. These findings provide a novel theoretical basis for understanding PE pathogenesis and highlight potential therapeutic targets for intervention.
Breast cancer remains a significant global health challenge, necessitating the development of innovative therapeutic strategies. This study aims to integrate immunotherapy and suicide gene therapy for breast cancer treatment. The cytokines IL-2 and GM-CSF, which are crucial in orchestrating immune responses against cancer, are the focal points of investigation. This study focused on the co-expression of these cytokines alongside apoptin, a promising protein derived from chicken anemia virus, via the bidirectional survivin promoter (pAIG). The constructed vectors were transfected into MCF-7 cells, which resulted in a significant increase in gene expression. Compared with individual cytokine and apoptin vectors (pIG and pA), the pAIG vector exhibited superior efficacy, reducing cell viability and inducing apoptosis (31.7%) at 72 h post-transfection. Exploration of the interaction of apoptin revealed its association with the human PIK3R1 protein, which interacts with both cytokines, contributing to the inhibition of cell viability. Leveraging the survivin promoter for co-expression presents a promising avenue for targeted breast cancer gene therapy, potentially disrupting intricate signaling networks involved in cancer progression. This study provides valuable insights into the synergistic effects of IL-2, GM-CSF, and apoptin co-expression, offering a compelling approach to advancing breast cancer treatment. These findings contribute to the broader landscape of cancer gene therapy, emphasizing the potential of combining immunomodulatory agents and gene-directed approaches to improve therapeutic outcomes.
PROBLEM:The advent of high-throughput 16S rRNA sequencing has enabled deeper insights into microbial communities associated with adverse pregnancy outcomes, including spontaneous preterm birth (sPTB). While microbial dysbiosis in the cervicovaginal and oral-gut microbiomes has been implicated in sPTB, the existence of a placental microbiome remains contentious. Traditional paradigms of a "sterile womb" have been challenged by studies suggesting a low-biomass microbial community in the placenta, though recent evidence disputes this claim, attributing findings to contamination or transient microbial DNA signals. METHOD:This study systematically reviewed placental microbiome studies employing 16S rRNA sequencing and re-analyzed publicly available datasets to determine microbial signatures in term and preterm placentas. Following a comprehensive search of three databases and stringent inclusion criteria, seven studies were included. The risk of bias was assessed using a modified Joanna-Briggs tool, revealing moderate-to-low risk across studies. Methodological heterogeneity, including differences in contamination controls, sequencing regions, and analytical platforms, was a significant limitation. RESULTS:A re-analysis of sequencing data showed no consistent microbiome signature distinguishing the term from preterm placentas. Beta diversity analysis revealed no group clustering, while alpha diversity indices showed comparable species richness. Bacterial DNA in placental tissues was primarily attributed to contamination from the urogenital tract or laboratory processes. CONCLUSION:Findings underscore the importance of robust contamination control and standardized protocols in low-biomass microbiome research. Future studies should employ advanced techniques, such as metagenomics and fluorescence in situ hybridization, to evaluate the functional relevance of microbial communities in the placenta, as well as rule out microbial DNA deposited in the placenta through circulating bacterial extracellular vesicles (EVs).
OBJECTIVE:The pathophysiological mechanisms underlying second trimester pregnancy loss are not yet fully elucidated. This study aimed to evaluate whether maternal serum gelsolin levels, together with hematological inflammatory indices and pregnancy related hormones, are associated with second trimester pregnancy loss. METHODS:A total of 912 pregnant women were prospectively enrolled, and a nested matched case-control analysis was subsequently conducted including 112 participants (40 cases and 72 matched controls). The study was conducted at Ankara Etlik City Hospital between January and December 2024. The pregnancy loss group comprised 40 women who experienced second-trimester pregnancy loss, whereas the control group included 72 age- and body mass index-matched women with ongoing pregnancies. Maternal inflammatory markers-including the neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), systemic inflammatory response index (SIRI), systemic immune-inflammation index (SII), and aggregate index of systemic inflammation (AISI)-were evaluated. In addition, serum levels of human chorionic gonadotropin (hCG), pregnancy-associated plasma protein-A (PAPP-A), and gelsolin were measured and compared between groups. RESULTS:Most hematological inflammatory indices, including NLR, PLR, MLR, SII, SIRI, and AISI, did not differ significantly between the groups. However, maternal serum PAPP-A levels were significantly lower in the pregnancy loss group (p = 0.012), whereas gelsolin levels were significantly higher (p = 0.034). Receiver operating characteristic (ROC) analysis demonstrated limited discriminatory performance for both markers. PAPP-A MoM showed an AUC of 0.644 (95% CI: 0.533-0.755, p = 0.012), with a cut-off value of <0.46 yielding a sensitivity of 77.5% and specificity of 91.7%. Despite its high specificity at the selected cut-off, overall discriminatory performance was limited. Gelsolin exhibited modest diagnostic performance with an AUC of 0.621 (95% CI: 0.501-0.741, p = 0.034), with a sensitivity of 57.5% and specificity of 44% at a threshold of >0.893 ng/mL. CONCLUSION:Maternal serum gelsolin levels were elevated and PAPP-A levels were reduced in pregnancies affected by second-trimester pregnancy loss. While gelsolin remained independently associated with pregnancy loss after adjustment, neither marker demonstrated sufficient discriminatory performance to be used as a standalone predictive tool. These findings may reflect underlying placental dysfunction and inflammatory dysregulation rather than overt systemic inflammation, and should be considered hypothesis-generating pending validation in larger prospective studies.
PROBLEM:Female genital mutilation/cutting (FGM/C) is harmful to physical, mental, and reproductive health, though the effect of this practice on a woman's HIV susceptibility is poorly understood. Despite the known associations of FGM/C with short-term vaginal epithelial damage, neither genital inflammation nor the genital microbiome have been explored in women who have undergone FGM/C. In this study we compare the genital immune milieu and microbiome among female sex workers (FSWs) by FGM/C status, hypothesizing that these biological factors are dysregulated in women who have undergone FGM/C, heightening their risk of HIV acquisition. METHOD OF STUDY:1003 FSWs in Nairobi, Kenya, were enrolled in the Maisha Fiti study and visited a study clinic up to three times from June 2019 to March 2021. Participants self-reported any previous exposure to FGM/C as well as other relevant sociodemographic factors. Levels of proinflammatory cytokines and soluble E-cadherin (sE-cad), a biomarker of epithelial barrier disruption, were measured by multiplex immunoassay using self-collected cervicovaginal secretion samples provided by HIV-uninfected participants. Genital inflammation was defined using a composite score of inflammatory cytokines previously associated with HIV acquisition. The presence of inflammation was compared longitudinally between groups using mixed models to control for potential confounders including age, bacterial vaginosis (BV) status as defined by Nugent score, and others. Vaginal bacterial abundance, Shannon diversity, and total levels of key vaginal bacteria were measured by qPCR and compared by FGM/C status in an exploratory analysis. RESULTS:44 of 1003 (4%) participants had undergone Type I or II FGM/C. These participants were older (p < 0.001) and more likely to test positive for herpes simplex virus-2 (HSV-2; p = 0.04), and less likely to have completed primary education (p = 0.03). Among HIV-uninfected participants, there was no evidence that genital inflammation was associated with FGM/C status after controlling for potential confounders (aOR = 0.70; 95% CI: 0.31-1.59; p = 0.40). There was no evidence of a difference in BV prevalence (p > 0.99), total bacterial abundance (p = 0.96), or Shannon diversity (p = 0.15) by FGM/C status. CONCLUSIONS:Type I or II FGM/C was not associated with genital inflammation or microbial dysregulation in the long-term among HIV-negative FSWs in this cohort. This may be due to the duration elapsed since FGM/C occurred or the lowered mucosal immune activation previously observed in FSWs.
OBJECTIVE:This study aims to explore how recombinant human granulocyte colony-stimulating factor (rhG-CSF) impacts pregnancy outcomes among patients with recurrent pregnancy loss (RPL), especially the effects of different rhG-CSF administration types on these outcomes. METHODS:This retrospective cohort study analyzed 504 RPL patients treated at the Lanzhou University Second Hospital, including 291 in the treatment group (receiving rhG-CSF) and 213 in the control group. Potential confounding factors were adjusted by Propensity Score Matching (PSM) and Inverse Probability of Treatment Weighting (IPTW). Logistic regression was employed to assess the association between rhG-CSF and pregnancy outcomes. Additionally, exploratory subgroup analysis was performed based on different rhG-CSF administration types (short-acting, long-acting, and sequential therapy). RESULTS:Overall, rhG-CSF treatment did not significantly improve the rates of live births (OR = 1.259, 95% CI: 0.771-2.062, p = 0.357), preterm births (OR = 1.095, 95% CI: 0.304-6.143, p = 0.917), or neonatal unit admissions (OR = 1.123, 95% CI: 0.205-7.511, p = 0.898). However, exploratory subgroup analysis revealed that short-acting rhG-CSF was linked to a lower live birth rate (LBR) (aOR = 0.476, 95% CI: 0.224-0.971), while sequential therapy significantly increased LBRs (aOR = 2.417, 95% CI: 1.384-4.268). Further analysis showed that there were no significant differences in the rates of preterm birth and neonatal unit admissions among different groups. CONCLUSION:Although rhG-CSF showed no significant overall benefit for pregnancy outcomes in RPL patients, exploratory subgroup analyses revealed divergent effects based on administration types. While a potential negative association was observed with short-acting rhG-CSF and a positive impact with sequential therapy, these subgroup-specific findings should be interpreted with caution and require validation in larger, dedicated prospective cohorts.
PROBLEM:Pregnancy is characterized by significant changes in peripheral immunity. Total CD3+ T cells decrease across pregnancy while Forkhead Box P3+ T-regulatory cells (FoxP3+Tregs) peak in mid-pregnancy, the latter of which are key to healthy outcomes. However, less is known regarding distinct memory populations of FoxP3+ Tregs and their phenotypic changes across pregnancy. METHOD OF STUDY:Pregnant individuals were enrolled into a prospective cohort study and high parameter flow cytometry was used to characterize peripheral blood mononuclear cells collected at first and second trimester and delivery. Memory and phenotypic marker expression on FoxP3+ Tregs at each timepoint were analyzed. Non-Treg CD4+ T cells and CD8+ Tcells were assessed as comparison populations. Suppressive capacity of FoxP3+ Tregs was compared between first trimester and delivery. RESULTS:The frequency of central and effector memory populations within FoxP3+ Tregs, non-Treg CD4+ T cells, and CD8+ T cells decreased across pregnancy (p ≤ 0.001 for all). Inducible T cell Costimulator (ICOS) expression decreased on FoxP3+ Tregs across pregnancy (p = 0.01), while T cell immunoglobulin and mucin domain-containing protein 3 (Tim3) expression increased on FoxP3 Tregs, non-Treg CD4+, and CD8+ T cells (p ≤ 0.005 for all). Suppressive capacity of Tregs did not vary by gestational timepoint. CONCLUSIONS:We observed an increase in naïve cells in FoxP3+ and non-Treg CD4+ T cells across gestation and a corresponding reduction in ICOS expression. However, the observed increase in Tim3 expression on all T cell subsets suggests a dissociation between suppressive markers in pregnancy. Together, these data suggest that pregnancy has a unique impact on Treg memory distribution and phenotype.
BACKGROUND:Preeclampsia (PE) is a major pregnancy complication that poses risks to both the mother and the fetus. Oxidative stress (OS) plays a crucial role in its pathogenesis. This study aims to explore the diagnostic value of oxidative stress-related genes for PE. METHODS:We integrated single-cell and batch transcriptome data from the GEO database. Through differential expression analysis, WGCNA, and three machine learning algorithms (LASSO, SVM-RFE, and random forest), we screened key genes, constructed a diagnostic model, and analyzed its correlation with immune cells. We used bioinformatics tools to predict regulatory networks. The diagnostic value of key genes was verified through qRT-PCR experiments. RESULTS:Our study identified three hub genes-SPP1, CTSC, and SLC9A9-which were predominantly highly expressed in macrophages. The Hub gene and diagnostic model showed preliminary discriminatory ability in distinguishing PE samples from control samples in multiple datasets. Immune infiltration analysis demonstrated a strong positive correlation between macrophages and the three hub genes. Furthermore, cell adhesion emerged as a key signaling pathway in PE progression. FOXC1 was a common transcriptional regulator of all three hub genes, while FOXL1, NFKB1, and POU2F2 co-regulate SLC9A9 and SPP1. qRT-PCR validation confirmed the downregulation of these hub genes in PE. CONCLUSION:This study successfully identified SPP1, CTSC, and SLC9A9 as candidate biomarkers for PE, which may lay the foundation for the development of new monitoring indicators and optimization of targeted therapy strategies.
PROBLEM:Immunomodulatory agents (IVIG, intralipid, corticosteroids, and G-CSF) and platelet-rich plasma (PRP) are increasingly used in infertility despite inconsistent clinical outcomes. A critical but overlooked question is whether these agents exert their effects through systemic immune modulation or require direct access to target cells within the ovary and endometrium. HYPOTHESIS:We propose that insufficient tissue accessibility, rather than intrinsic biological inefficacy, may contribute to inconsistent clinical outcomes. This framework examines the physical constraints governing therapeutic effectiveness if local action were necessary, while acknowledging that benefits may also be mediated systemically. FINDINGS:Anatomical and biophysical barriers differ by administration route. For systemic agents, the vascular endothelium (continuous capillaries) restricts macromolecular extravasation via capacity-limited transcytosis. For local agents, epithelial tight junctions (0.5-2 nm pores), follicular basal lamina (40-100 nm), granulosa cell layers (∼20 nm), and the zona pellucida (5-10 nm) create sequential barriers. Molecular size analysis suggests that large immunomodulators like IVIG (∼150 kDa, 10-15 nm) and intralipid particles (200-500 nm) face substantial limitations in reaching target compartments. PRP growth factors face identical barriers. By contrast, smaller molecules (<1 kDa) and FSH (∼30 kDa, 5 nm) penetrate more readily. Ovarian tissue stiffness (1.5-5 kPa in normal ovaries; 0.9-11.3 kPa in premature ovarian insufficiency) further impedes macromolecular diffusion. The proposed Mathematical Access Index integrates molecular size, barrier pore dimensions, and tissue stiffness to predict accessibility. CONCLUSION:Accessibility of immunomodulators and PRP growth factors to reproductive tissues is more limited than commonly assumed. If these therapies act predominantly through systemic immune modulation (e.g., Fc receptor saturation, Th1/Th2 modulation), direct tissue penetration may not be necessary. For PRP, observed effects likely result from indirect mechanisms (paracrine signaling, angiogenesis). Distinguishing between systemic and local mechanisms is essential. Until pharmacokinetic studies quantify drug concentrations in reproductive tissues, clinical use should remain cautious.
OBJECTIVE:To determine whether plasma acute phase-, adhesion-related, and multifunctional proteins, alone or in combination with ultrasound and traditional blood tests, can predict the microbial invasion of the amniotic cavity and/or intra-amniotic inflammation (MIAC/IAI) and spontaneous preterm birth (SPTB) in women with preterm labor (PTL). STUDY DESIGN:A retrospective cohort study was performed on 249 singleton pregnant women with PTL (at 23+0 to 34+0 weeks) who underwent transabdominal amniocentesis. Plasma samples were obtained at amniocentesis. MIAC/IAI was characterized by a positive amniotic fluid (AF) culture and/or an elevated interleukin (IL)-6 level (≥2.6 ng/mL) in the AF. Plasma levels of hepcidin, mannose-binding lectin (MBL), retinol-binding protein 4 (RBP4), serum amyloid A1 (SAA1), E-selectin, P-selectin, progranulin, transforming growth factor β1, and vitamin d-binding protein levels were measured using enzyme-linked immunosorbent assay. C-reactive protein levels, white blood cell (WBC) counts, neutrophil-to-lymphocyte ratio, and sonographic cervical lengths (CLs) were measured. RESULTS:After adjusting for baseline covariates, multivariable logistic regression analyses showed significant associations between (i) decreased RBP4 and E-selectin levels and elevated SAA1 levels in the plasma and MIAC/IAI; and (ii) decreased RBP4 and progranulin levels and elevated SAA1 levels in the plasma and SPTB ≤7 days of sampling. Using stepwise regression analyses, noninvasive prediction models for MIAC/IAI and SPTB ≤7 days were developed, comprising plasma RBP4, SAA1, MBL, and E-selectin levels; CL; and WBC counts, with area under the curve values of 0.86 and 0.89, respectively. CONCLUSIONS:SAA1, RBP4, E-selectin, and progranulin may represent valuable plasma biomarkers for predicting MIAC/IAI and imminent SPTB in women with PTL. Particularly, the combination of these plasma biomarkers with ultrasound and traditional inflammatory blood markers can significantly support the diagnosis of MIAC/IAI and imminent SPTB.
OBJECTIVE:To screen immune-related biomarkers in diagnosing patients with both endometriosis (EM) and systemic lupus erythematosus (SLE). METHODS:After performing differential expression analysis, immune infiltration analysis, WGCNA, the immune-related genes in EM and SLE were screened. Then the diagnostic genes were identified by three machine learning algorithms, followed by evaluation of the predictive performance of the diagnostic genes by nomogram and ROC curve. Then the correlation between diagnostic genes and immune cells, TFs, GSEA, and potential drugs prediction analyses were performed. Lastly, experiments in vitro were applied to explore the function of SUPT16H in EM and SLE. RESULTS:Total 20 immune-related genes in EM and SLE were identified by intersecting DEGs and module genes. Using "LASSO", "RF", and "SVM-RFE" algorithms, and total three common diagnostic genes were obtained, namely, C1QC, SOCS3, and SUPT16H. ROC curve shown that AUCs of diagnostic genes were all above 0.7 in training and verification datasets. The targeted drugs for the three diagnostic genes were predicted, containing Pingyangmycin CTD 00001211, VANADIUM PENTOXIDE CTD 00002655, CTD 00001728, and so forth. Also, SUPT16H exerted significant function in occurrence of EM and SLE via regulating inflammation and oxidative stress in cell experiments in vitro. CONCLUSION:SUPT16H overexpression alleviates the progression of EM and SLE by inhibiting inflammation and oxidative stress. The three co- susceptibility genes (C1QC, SOCS3, and SUPT16H) that strongly related to immunity in EM and SLE could be the promising candidate biomarker for the diagnosis and treatment for EM and SLE patients.
PROBLEM:Embryo implantation failure remains a significant challenge in reproductive medicine. This study investigates the therapeutic potential and underlying mechanism of a Chinese medicine Bushen Huoxue Formula (BHF) enema in improving embryo implantation. METHOD OF STUDY:A rat model of embryo implantation disorder was established, with uterine tissues collected on the sixth and eighth days post coition (dpc). Implanted embryo counts were recorded, and endometrial morphology and pinopode development were assessed using hematoxylin-eosin staining and scanning electron microscopy. Western blotting and immunofluorescence measured lipoxin A4 receptor (LXA4R), nuclear factor kappa-B (NF-κB) p65, and epithelial-mesenchymal transition (EMT) marker expressions. An in vitro model was also used, where adhesion assays, Western blotting, and RT-qPCR were conducted before and after NF-κB p65 silencing via lentiviral transfection to evaluate BHF enema-containing serum (BHFE-CS) effects. RESULTS:BHF enema significantly increased endometrial thickness, improved pinopode development, and enhanced the number of implanted embryos in model rats. Molecularly, BHF upregulated p-NF-κB p65 and N-cadherin while downregulating LXA4R and E-cadherin, promoting a pro-inflammatory microenvironment and EMT process essential for embryo implantation. Consistently, BHFE-CS enhanced embryo adhesion in vitro. Crucially, the pro-implantation effects of BHFE-CS were abolished upon NF-κB p65 silencing, confirming the pathway's indispensability. CONCLUSIONS:BHF enema improves embryo implantation by modulating the LXA4R/NF-κB pathway to promote EMT, highlighting its potential as a treatment for embryo implantation disorders.
Objective Postmenopausal bleeding (PMB) and increased endometrial thickness are key clinical indicators that may suggest underlying malignancies. While endometrial biopsy remains the diagnostic gold standard, its invasiveness underscores the need for alternative, noninvasive biomarkers. This study evaluates the potential of clinical, laboratory, and peripheral blood inflammatory indices (PBII) in distinguishing malignant from benign endometrial pathologies.Methods This retrospective study included 162 patients who underwent endometrial biopsy due to PMB and/or increased endometrial thickness between January 2023 and January 2024. Patients were categorized into benign (n = 134) and malignant (n = 28) groups. Demographic, clinical, and laboratory parameters were collected, PBII parameters were calculated, and comparisons were performed. Logistic regression analyses were conducted to identify independent predictors of malignancy.Results Malignant cases were significantly associated with older age (p < 0.001), longer postmenopausal duration (p = 0.002), higher body mass index (BMI) (p = 0.018), and greater endometrial thickness (p = 0.042) compared to benign cases. Hemoglobin levels were significantly lower (p = 0.022), while neutrophil (p < 0.001) and monocyte (p = 0.042) counts were notably higher in malignant cases. Among PBII parameters, neutrophil-to-lymphocyte ratio (NLR), systemic immune-inflammation index (SII), pan-immune-inflammation value (PIV), and systemic inflammation response index (SIRI) were significantly elevated (p < 0.001 for all). Multivariate analysis identified older age (p < 0.001), lower hemoglobin (p = 0.016), higher neutrophil count (p = 0.030), and increased PIV (p = 0.022) as independent predictors of malignancy.Conclusion Integrating clinical and laboratory parameters with PBII, particularly PIV, may be a valuable, noninvasive tool for the early detection and risk stratification of endometrial malignancies. This approach could enhance diagnostic accuracy, reduce the need for invasive biopsies, and improve patient management.
Problem Maternal obesity is associated with elevated inflammatory markers, hormonal dysregulation, and metabolic disturbances. However, how maternal obesity modulates systemic inflammatory, hormonal, and redox profiles in the peripartum and postpartum periods remains incompletely understood.Method of Study This observational study evaluated inflammatory biomarkers, steroid hormone levels, and oxidative stress markers in obese (n = 8) and non-obese (n = 11) pregnant women undergoing labor induction. Peripheral maternal blood samples were collected immediately before induction and again within approximately 5-10 mins after delivery. Placental tissue was collected postpartum within the same interval. Comparative analyses between groups and time points were performed using repeated-measures statistical models.Results C-reactive protein (CRP) levels increased postpartum only in the obese group, indicating an enhanced inflammatory response after delivery. In contrast, interleukin-6 (IL-6) levels declined postpartum only in the non-obese group. No between-group differences were detected in the oxidative stress markers assessed, either in maternal blood or placental tissue.Conclusions Maternal obesity is associated with distinct postpartum inflammatory and hormonal profiles, characterized by sustained CRP elevation and altered cytokine dynamics. No between-group differences were detected in the redox markers assessed, suggesting preserved redox balance in the parameters evaluated. These findings highlight the importance of considering postpartum immune modulation in obese parturients and support further investigation into obesity-associated inflammatory regulation during the peripartum period.
PROBLEM:How does the traditional Chinese compound Bushen Xiaozheng Decoction (BSXZD) affect the immunosuppressive microenvironment in rat endometriosis models? METHODS:Sprague Dawley rats were randomly divided into normal, sham operation, model, Yasmin, and low-, medium-, high-dose BSXZD groups with 10 rats in each group. The endometriosis model was established by autologous endometrial transplantation, and corresponding interventions were given for 28 days. The volume of ectopic lesions was measured, and histological changes of endometrium were observed by hematoxylin-eosin staining. The protein or gene expression of interleukin-10 (IL-10) and transforming growth factor-β (TGF-β) in serum or endometrium was detected by immunohistochemistry, quantitative real-time polymerase chain reaction, Western blotting, and enzyme-linked immunosorbent assay. RESULTS:Following treatment with BSXZD, the pseudostratified phenomenon and cavity structure of endometrial cells were reduced. Additionally, the expression levels of IL-10 and TGF-β in both endometrium and serum were significantly decreased, accompanied by a marked reduction in the volume of ectopic lesions. CONCLUSION:BSXZD can significantly inhibit the growth of ectopic lesions in a rat model of endometriosis. The underlying mechanism may involve dual regulation of the endocrine-immune axis: it not only directly downregulates the expression of the immunosuppressive factors IL-10 and TGF-β to reversing the local immunosuppressive microenvironment, but also indirectly modulates these cytokines via sex hormone-related signaling pathways. Further studies are needed to clarify the precise molecular mechanism.
