A current working hypothesis in recurrent pregnancy loss (RPL) suggests immune dysregulation, underlain by a pro-inflammatory cytokine shift in TH cells. NK cells, which are primary immune effectors throughout pregnancy, also orchestrate immune activity via cytokine secretion, which appears aberrant in women with RPL. Experimental findings remain contentious, hindering clinical adoption of cytokine testing, despite increasing pressure from clinicians and couples affected. This study aims to identify peripheral cytokine biomarkers that could be utilized for the stratification of women with RPL displaying underlying immune dysregulation. Peripheral blood samples from non-pregnant women with RPL (n = 99) and controls (n = 80) were assessed for TH and NK cell cytokine production via flow cytometry. Cytokine secretion by peripheral blood mononuclear cells was examined via ELISA. Elevated pro-/anti-inflammatory cytokine-producing TH cell ratios were detected in RPL (P < 0.0001). This was, however, accompanied by reduced pro-/anti-inflammatory cytokine expression within individual cells (IL-2/IL-10: P < 0.0001, IFN-γ/IL-10: P < 0.0001, TNF-α/IL-10: P = 0.0136, IL-17A/IL-10: P < 0.0001). NK cell cytokine production appeared deficient overall in women with RPL (TNF-α+ NKs: P < 0.0001, IL-10+ NKs: P = 0.0006). Interestingly, a pro-inflammatory shift was evident when cytokine secretion was investigated (TNF-α/IL-5: P = 0.0120, IL-2/IL-13: P = 0.0105, IFN-γ/IL-13: P = 0.0040, TNF-α/IL-13: P < 0.0001). A pro-inflammatory shift in cytokine secretion was identified in women with RPL, indicating systemic cytokine dysregulation. Peripheral cytokine levels emerged as discriminatory and, thus, upon validation, may present valuable biomarkers in the identification of immune-associated RPL. Further research on the functional effects of cytokine imbalance in utero is required to strengthen the pathogenic relevance of these findings.

Recurrent spontaneous abortion (RSA) involves complex and often unexplained pathogenesis. This study investigated metabolic reprogramming in RSA to identify diagnostic biomarkers. We integrated GEO datasets (GSE26787, GSE165004), identified metabolic reprogramming-related differentially expressed genes, and intersected them with WGCNA modules. Module genes were refined using logistic regression, SVM-RFE, and LASSO to construct a diagnostic model validated by ROC, nomogram, calibration, and decision curve analysis. The role of SREBF2 was explored by PCR, western blot, CCK-8, and transwell assays in vitro. Immune infiltration (ssGSEA, CIBERSORT) and single-cell RNA sequencing (GSE214607) explored immune-metabolic crosstalk. Results demonstrate critical involvement of metabolic reprogramming in RSA. Four model genes (SREBF2, PPARG, SQLE, UCP2) were upregulated in RSA and enriched in lipid/cholesterol pathways. Experimental validation confirmed SREBF2 upregulation in RSA decidua. Its overexpression in endometrial stromal cells impaired cell proliferation, decidualization, and trophoblast migration. Immune infiltration revealed correlations between model genes and specific immune subsets. Single-cell analysis identified altered immune cell proportions and cell-type-specific expression patterns, predominantly in macrophage subsets. This study indicates that lipid-centric metabolic remodelling may concurrently affect decidual function, trophoblast behaviour, and immune tolerance in RSA, offering diagnostic and therapeutic insights.

ObjectiveUnexplained recurrent spontaneous abortion is a multifactorial condition with an unclear etiology. This study aimed to investigate the relationship between liver function biomarkers and unexplained recurrent spontaneous abortion and explore interactions with tobacco exposure and body mass index.MethodsA retrospective case-control study was conducted on 2126 pregnant women, including 602 with unexplained recurrent spontaneous abortion and 1524 who served as controls. Liver enzyme indicators (alanine aminotransferase, aspartate aminotransferase, gamma-glutamyl transferase, and alkaline phosphatase) were assessed, and multivariate logistic regression models were applied to analyze their association with unexplained recurrent spontaneous abortion. Interaction analyses were performed to evaluate combined effects of liver enzyme abnormalities, tobacco exposure, and body mass index.ResultsLiver enzyme abnormalities (alanine aminotransferase level >32 U/L or aspartate aminotransferase level >28 U/L) were significantly associated with increased unexplained recurrent spontaneous abortion risk (adjusted odds ratio = 2.15, 95% confidence interval: 1.68-2.76). A significant interaction between alanine aminotransferase elevation and tobacco exposure was observed (p < 0.001). Among non-exposed women, elevated alanine aminotransferase levels were associated with an odds ratio of 1.8 (95% confidence interval: 1.2-2.7), whereas among tobacco-exposed women, the odds ratio was 5.2 (95% confidence interval: 3.1-8.7) (p for interaction <0.001). However, these interaction results warrant cautious interpretation due to the absence of quantitative tobacco exposure metrics. The alanine aminotransferase-body mass index interaction was not significant (p = 0.179).ConclusionsElevated alanine aminotransferase and aspartate aminotransferase levels are independently associated with unexplained recurrent spontaneous abortion, and tobacco exposure significantly strengthened this association. These findings suggest that liver function abnormalities and tobacco exposure are associated with unexplained recurrent spontaneous abortion risk. However, given the limitations associated with tobacco exposure measurement, these findings should be interpreted with caution.

We report the case of a 37-year-old woman who presented with hyperemesis gravidarum at 11 weeks of gestation. Laboratory examination revealed severe thyrotoxicosis (TSH: <0.005 μIU/mL, FT3: 12.95 pg/mL, FT4: 3.93 ng/dL) with negative anti-TSH receptor antibody, while serum human chorionic gonadotropin (hCG) was markedly elevated at 198,983 mIU/mL. She was diagnosed with gestational transient thyrotoxicosis (GTT), although the presence of thyroid-stimulating antibody raised suspicion for concomitant Graves' disease (GD). Because of severe and clinically burdensome thyrotoxic symptoms, including excessive sweating and weight loss, propylthiouracil therapy was initiated, but thyrotoxicosis recurred after 20 weeks of gestation alongside elevated serum hCG levels, prompting a switch to thiamazole. Genetic testing revealed no TSHR mutations. Postpartum, both hCG and thyroid hormone levels normalized without treatment. During the second pregnancy, she experienced a miscarriage at nine weeks. During the third pregnancy, an elective abortion was performed due to a fetal genetic disorder. Both pregnancies were marked by severe thyrotoxicosis, consistent with GTT. During the fourth pregnancy, at 11 weeks, she developed hyperemesis gravidarum and was diagnosed with GTT. The persistently elevated thyroglobulin levels (>200 ng/mL; reference range:, <33.7 ng/mL) were also compatible with a possible contribution from painless thyroiditis (PT). Serum hCG levels normalized after the second trimester. Recurrent and sustained thyrotoxicosis during pregnancy across multiple pregnancies in the same patient is rare and requires careful differentiation among possible etiologies, including GTT, GD, and PT. This case underscores the importance of comprehensive monitoring of placental function, thyroid autoimmunity, and thyroiditis throughout pregnancy to ensure accurate diagnosis and tailored management for both mother and fetus.

In recent years, the role of the vaginal microbiota in pregnancy maintenance has garnered increasing attention. Extensive research indicates that vaginal dysbiosis is closely associated with various adverse pregnancy outcomes, including ectopic pregnancy, miscarriage, preterm birth, and premature rupture of membranes. Recurrent spontaneous abortion (RSA) affects approximately 1-5% of women of childbearing age and has a complex etiology, with infection implicated in about 4% of cases. This systematic review examines the compositional characteristics of the vaginal microbiota, its dynamic changes during pregnancy, and the underlying mechanisms linking it to RSA. It focuses on the protective role of lactobacilli, ascending pathogen infection, and local immune regulation, aiming to provide a novel theoretical foundation for the prevention, risk prediction, and personalized treatment of RSA.

Spontaneous abortion is one of the most common adverse pregnancy outcomes, with an incidence of approximately 15-25% in clinically recognized pregnancies. Chromosomal abnormalities are recognized as a primary genetic etiology, yet the specific underlying molecular mechanisms and pathways remain incompletely understood. Embryonic tissue samples from 1585 patients with spontaneous abortion were collected from January 2019 to June 2025. Chromosomal microarray analysis (CMA) was used for whole-genome detection of chromosomal abnormalities, including aneuploidy and copy number variations (CNVs). Functional enrichment analysis of Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were conducted on CNV-related genes. 34 cases with negative CMA results and a history of recurrent pregnancy loss (≥ 2 episodes) were randomly selected to undergo whole-exome sequencing (WES) for the screening of monogenic variants. Statistical analysis included chi-square tests for inter-group comparisons, restricted cubic splines (RCS), and multivariate logistic regression for assessing risk factors. The detection rate of chromosomal abnormalities was 54.44%, mainly numerical abnormalities, with trisomy 16 and X monosomy being the most common subtypes. Structural variants include CNVs, loss of heterozygosity (LOH) and Runs of homozygosity(ROH). The results of the GO enrichment analysis indicated that genes associated with CNVs are primarily involved in hemostasis regulation and monocyte migration; KEGG enrichment analysis indicated that the Toll-like receptor signaling pathway and the neuroactive ligand-receptor interaction pathway may play a significant role in the pathogenesis of miscarriage. Multivariate Logistic regression analysis revealed that a mother's age of ≥ 35 years was an independent risk factor (OR = 1.72, 95%CI: 1.31-2.26), while a gestational age of ≥ 12 weeks had a protective effect (OR = 0.27, 95%CI: 0.21-0.34). The RCS model revealed a U-shaped relationship between maternal age and the risk of chromosomal abnormalities. The risk was lowest at ages 28-29 (OR = 0.99) and increased gradually after age 30; the risk of chromosomal abnormalities peaked between 8 and 10 weeks of gestation. The diagnostic yield of WES in recurrent miscarriage cases with normal CMA results reached 26.47%. This approach identified a variant in the MOS gene associated with oocyte maturation defects, along with clinically significant variants in possible miscarriage-implicated genes such as TTC7A. Chromosomal abnormalities, particularly trisomy 16 and monosomy X, along with dysregulation of hemostatic and immune pathways, are central to spontaneous abortion etiology. Advanced maternal age, defined as 30 years or older, and early gestational loss are key risk factors, challenging the conventional threshold of 35 years. The use of a combined CMA and WES strategy has effectively improved the genetic diagnosis rate, demonstrating the complementary value and advantages of combining chromosomal and single-gene testing technologies in diagnosing the causes of miscarriage.