Extravillous trophoblasts (EVT) are essential for placental invasion into the maternal decidua and spiral artery remodeling, ensuring a successful pregnancy. It is well-established that early placental development occurs under physiologically hypoxic conditions (~2% O2), while the later stages proceed under increased physiological oxygen levels (~8% O2). However, the heterogeneity of EVT across these distinct developmental conditions remains poorly characterized. Moreover, the molecular mechanisms governing EVT lineage development and function under low oxygen conditions remain largely elusive. Herein, we systematically characterize the transcriptome dynamics underlying the differentiation of cytotrophoblasts (VCT) and human trophoblast stem cells (hTSC) into EVT at single-cell resolution. Our analysis reveals pronounced functional and phenotypic heterogeneity among EVT from early versus late gestational stages, as well as between in vivo and in vitro models. We demonstrate that oxygen tension is a pivotal factor driving EVT heterogeneity, giving rise to distinct subtypes with invasive/migratory and secretory potentials. Furthermore, we elucidate that hypoxia impairs EVT lineage development from hTSC in vitro. Hypoxia reduces intracellular UDP-N-acetyl-d-glucosamine (UDP-GlcNAc) level, thereby possibly disrupting protein N-glycosylation. We further demonstrate that N-glycosylation pathway activity is significantly elevated during the differentiation of hTSC into EVT, as pharmacological inhibition with tunicamycin (TM) completely blocks this process. UDP-GlcNAc supplementation rescues the EVT differentiation defect under hypoxia, demonstrating that protein N-glycosylation is essential for EVT lineage commitment under low oxygen conditions. Our findings uncover a hypoxia-sensitive glycosylation in EVT development and provide new insights into early placental development.

Environmental toxicant exposure has emerged as a potential contributor to diabetes, yet systematic investigations integrating multiple chemicals and biological mechanisms remain limited. This study employed a hypothesis-generating, exposome-toxicogenomic framework to examine the associations between diverse environmental toxicants and diabetes risk and to explore underlying biological pathways. Data from 2689 NHANES 2013-2016 participants (366 with diabetes and 2323 without) were analyzed. Forty-six toxicants across seven chemical classes were evaluated using exposure-wide association studies, deletion/substitution/addition modeling, restricted cubic splines, Bayesian kernel machine regression, and quantile-based g-computation. Integrative bioinformatics analyses, including Comparative Toxicogenomics Database annotations, transcriptomic profiling, pathway enrichment, protein-protein interaction networks, and machine learning, were conducted to explore potential biological pathways and support biological plausibility. Five toxicants-glycidamide, ethylene oxide, antimony, uranium, and NAC-3HPM-were consistently associated with higher odds of diabetes (OR range: 1.22-1.34). Mixture analyses revealed cumulative risk amplification (qgcomp OR 1.39, 95 % CI 1.21-1.60), with ethylene oxide showing the highest posterior inclusion probability (>0.5). Stronger associations were observed among obese individuals. Bioinformatics analyses identified 87 overlapping toxicant-diabetes-related genes enriched in pathways related to oxidative stress, apoptosis, AGE-RAGE signaling, and atherosclerosis. Machine learning across 113 models (optimal: Elastic Net; training AUC 0.956, test AUC 0.867) highlighted 14 key genes, of which five (MAPK8, SIRT1, PIK3R1, KRAS, MAPK1) overlapped as hub genes in protein-protein interaction networks. These findings suggest that background-level exposure to environmental toxicants is associated with increased diabetes risk, potentially involving biologically relevant pathways related to mitochondrial function, insulin signaling, and inflammatory processes, with obesity acting as a potential susceptibility factor.

Gestational diabetes mellitus (GDM) represents a prevalent pregnancy complication with long-term health implications for offspring. While metabolic outcomes have been extensively studied, sex-specific effects on neurodevelopment remain poorly understood. Here we investigated the sex-dependent impact of maternal GDM on offspring brain development and behavior using a high-fat diet and low-dose streptozotocin induced mouse model. We found that adult female offspring exposed to maternal GDM exhibited depressive-like behaviors and sustained impairments in hippocampal neurogenesis across multiple developmental stages (embryonic, weaning and adult), characterized by reduced neural stem cell proliferation and altered differentiation. By contrast, male offspring displayed substantial metabolic dysfunction but no sustained neurogenic deficits beyond the embryonic period. Metabolomic analysis revealed persistent downregulation of myo-inositol in female offspring hippocampus, associated with disruptions in neurogenic signaling pathways. In vitro experiments with female-derived neural stem cells confirmed that hyperglycemic conditions directly impaired proliferation and differentiation, partly through oxidative stress mechanisms. These findings establish a sex-specific vulnerability to GDM-induced neurodevelopmental alterations and identify myo-inositol metabolism as a potential therapeutic target for preventing long-term neuropsychiatric consequences in female offspring.Maternal GDM induces sex-specific effects on offspring neurodevelopment, with females exhibiting persistent hippocampal neurogenesis deficits and depressive-like behaviors, while males show neurogenic resilience. The identification of myo-inositol depletion and oxidative stress as potential contributors to female-specific neurogenic impairments provides new insights into sex-specific vulnerability to maternal metabolic disturbances and suggests potential targets for intervention. HFD, High fat diet; STZ, Streptozotocin; GCL, Granule cell layer; SGZ, Subgranular zone; NSCs, Neural stem cells. Figure created with BioRender.com.

Preeclampsia (PE) is a pregnancy-specific syndrome driven by placental dysfunction, and premature placental senescence has increasingly been implicated in its pathogenesis. However, the upstream regulatory mechanisms remain poorly understood. Here, we found that placental SIRT1 expression was reduced in PE and was accompanied by senescence-associated features. In trophoblasts, SIRT1 knockdown enhanced senescence and senescence-associated secretory phenotype (SASP) release, while impairing proliferative capacity, migration, and invasion, whereas pharmacological activation of SIRT1 attenuated placental senescence and ameliorated PE-like manifestations in vivo. Mechanistically, iron chelation alleviated senescence, whereas senolytic intervention partially restored iron homeostasis, supporting a self-reinforcing interaction between iron dyshomeostasis and senescence. Collectively, these findings identify the SIRT1-p53 axis as an upstream regulator linking iron dyshomeostasis to placental senescence, support the existence of a ferro-aging-like pathogenic program in diseased placentas, and provide new insights into the molecular basis of placental dysfunction in PE.

Abstract Background Chronic endometritis (CE) has two principal subtypes—non-specific chronic endometritis (NSCE) and endometrial tuberculosis (ETB). Both can adversely affect embryo implantation. Although ETB is generally more severe, its low incidence has limited comparative data on assisted reproductive outcomes versus NSCE. A direct comparison of pregnancy outcomes following in vitro fertilization (IVF) or intracytoplasmic sperm injection-embryo transfer (ICSI-ET) between patients with ETB and those with NSCE would help clarify whether differential management strategies are warranted and provide more precise guidance for clinical practice. Methods In this single-center retrospective study, clinical data were collected from infertile patients who underwent IVF/ICSI-ET at West China Second University Hospital, Sichuan University, between January 2019 and December 2024. All patients were diagnosed via hysteroscopic endometrial biopsy. ETB was verified through histopathology along with positive nucleic acid testing for the Mycobacterium tuberculosis complex, whereas NSCE was diagnosed by histopathology in combination with CD138 immunohistochemical staining. Results A total of 409 patients were included: 27 in the ETB group and 382 in the NSCE group. No significant differences were observed between the two groups in age, duration of infertility, baseline FSH level, or number of embryos transferred (P > 0.05). The clinical pregnancy rate was comparable between the ETB group (51.9%) and the NSCE group (49.0%) (P > 0.05). However, the live birth rate was numerically lower in the ETB group (30.0%) than in the NSCE group (42.4%), though this difference was not statistically significant (P > 0.05). A significant difference was found in the miscarriage rate between the two groups with the ETB group having a higher miscarriage rate (P < 0.05). Multivariate regression analysis identified ETB as an independent risk factor for miscarriage after IVF/ICSI-ET. Conclusions In patients undergoing IVF/ICSI-ET, ETB is associated with a significantly higher risk of miscarriage compared to NSCE, despite similar clinical pregnancy rates. This underscores the need for distinct clinical management and counseling in ETB cases.

Objective:Fine particulate matter (PM 2.5) may impair follicular development; however, the roles of its chemical components and their time-specific effects remain unclear. This study examined how PM 2.5 and its major components are related to oocyte-related outcomes among women undergoing assisted reproductive technology (ART) and identified critical exposure windows. Methods:A total of 51,122 ART cycles were analyzed. Individual exposures to PM 2.5 and its components, sulfate (SO 4 2-), nitrate (NO 3 -), ammonium (NH 4 +), organic matter (OM), and black carbon (BC), were estimated for three periods: recent (0-3 months), distal (4-12 months), and cumulative (0-12 months). Associations between total, mature, and normally fertilized oocytes were assessed using negative binomial regression. Distributed lag non-linear models (DLNM) identified sensitive windows, and mixture models evaluated joint effects. Results:Higher PM 2.5 and component exposures were consistently associated with poorer oocyte outcomes, with stronger effects for distal exposure. Two sensitive windows, 1 month and 6-11 months before retrieval, were identified. Mixture analyses indicated SO 4 2- and NH 4 + as the dominant contributors. Conclusion:Exposure to PM 2.5 showed component-specific and time-dependent reproductive toxicity. Both short- and long-term exposure may reduce oocyte quantity and quality, highlighting the importance of improving air quality to support female reproductive health and ART success.

Inflammation has been linked to female fertility, yet its impact on ovarian reserve and response remains unclear. This study investigates the associations between systemic inflammatory markers, ovarian reserve—assessed by anti-Müllerian hormone (AMH) and antral follicle count (AFC)—and ovarian response, measured by the number of oocytes retrieved, in women undergoing assisted reproductive treatment (ART). A retrospective cohort study of 19,282 women evaluated systemic inflammatory markers, including systemic immune-inflammation index (SII), platelet-neutrophil product (PPN), platelet-to-lymphocyte ratio (PLR), and neutrophil-to-lymphocyte ratio (NLR), derived from neutrophil, platelet, and lymphocyte counts. Blood samples were collected on menstrual cycle days 2–4 for complete blood count analysis. Linear regression assessed associations between inflammatory markers and ovarian reserve (AMH, AFC) or ovarian response (oocytes retrieved). Restricted cubic splines tested nonlinear relationships, and stratified analyses identified sensitive subpopulations. Elevated first-trimester systemic inflammatory markers, including SII, PPN, PLR, and NLR, were significantly associated with reduced ovarian reserve and ovarian response. Adjusted models confirmed dose-response declines: highest vs. lowest quartiles showed significant reductions in AMH (e.g., SII β=-0.037, 95

Intrahepatic cholestasis of pregnancy (ICP) is a pregnancy-specific liver disorder characterized by elevated maternal bile acids and an increased risk of adverse fetal outcomes. Although placental dysfunction is a key contributor to ICP pathogenesis, the underlying mechanisms remain incompletely understood. Here, we combined a Sprague-Dawley rat model of ICP with spatial metabolomics to delineate region-specific metabolic alterations within the placenta. Multivariate analysis revealed distinct metabolic signatures between ICP and control placentas, with pronounced reprogramming in trophoblast-enriched regions. Differential metabolite and pathway analyses identified significant perturbations in glycerophospholipid metabolism, pyruvate metabolism, phospholipase D signaling, and the tricarboxylic acid cycle. Notably, phosphatidic acid (PA) was broadly elevated, whereas its downstream product phosphatidylinositol 4,5-bisphosphate (PIP2) exhibited spatially restricted accumulation in trophoblasts. Mechanistically, taurocholic acid exposure in HTR-8/SVneo cells induced upregulation of PIP5K1, leading to PIP2 accumulation and increased trophoblast apoptosis via disruption of the Bcl-2/Bax balance. Silencing of PIP5K1 restored PIP2 homeostasis and attenuated apoptosis. Importantly, these molecular alterations were recapitulated in placental tissues from ICP patients, confirming activation of the PIP5K1-PIP2 axis in vivo. Collectively, our findings identify a spatially resolved PA-PIP5K1-PIP2 lipid signaling cascade that links bile acid-induced metabolic stress to trophoblast apoptosis and placental dysfunction in ICP.

Trophoblast cell fusion is a vital developmental process that enables the formation of the multinucleated syncytiotrophoblast (STB), which plays a central role in placental function and maternal-fetal exchange. Fusion defects in this lineage are closely associated with pregnancy complications such as preeclampsia (PE) and fetal growth restriction (FGR). Although the fusion of trophoblasts is a highly coordinated event, it involves multiple interdependent steps, including transcriptional programming, membrane remodeling, cytoskeletal rearrangement, metabolic adaptation, and immune regulation. Recent studies have uncovered critical molecular mediators at each of these levels. Transcription factors such as Glial cells missing transcription factor 1(GCM1), Krüppel-like factor 6(KLF6), and Transcription factor EB(TFEB) govern differentiation timing; fusion proteins, including Syncytin-1/2 and Mfn2, facilitate membrane merger; polarity regulators and actin-associated proteins like Par6 and CNN3 organize cytoskeletal architecture; metabolic reprogramming, particularly a shift from oxidative phosphorylation to glycolysis, supplies energy and biosynthetic precursors; and immune modulators such as pregnancy-induced factor 1 and Interleukin-10(IL-10) ensure a permissive environment for fusion at the maternal-fetal interface. Epigenetic mechanisms, including DNA methylation and histone modifications, further fine-tune the expression of fusion-related genes. Alongside mechanistic discoveries, a wide range of experimental models has been developed to investigate trophoblast fusion in vitro. These include traditional monolayer cell lines (e.g., BeWo), primary human trophoblasts, placental explants, trophoblast stem cells, and trophoblast organoids. Each model system provides distinct advantages in recapitulating aspects of syncytialization and placental physiology. Moreover, the integration of multi-omics technologies-such as single-cell and spatial transcriptomics, proteomics, metabolomics, and epigenomics-has expanded our understanding of the spatiotemporal dynamics and molecular complexity underlying trophoblast fusion. Despite these advances, several key challenges remain unresolved, including the lack of models that fully recapitulate the structural and functional features of the human maternal-fetal interface and the limited understanding of posttranslational and spatiotemporal regulatory mechanisms. Addressing these gaps will be essential for translating basic insights into diagnostic and therapeutic innovations for placental diseases.

BACKGROUND:Although sensor arrays are powerful for discriminating glycosaminoglycans, the construction of a sensor array is usually cost and labour inefficient. With the purpose to acquire a high dimensional response data using only a limited number of dyes, the rational assembly of fluorescent dye molecules may provide an alternative approach. We envision that co-assembling different dyes within a supramolecular structure can trigger multiple photophysical processes, which should produce high dimensional optical signals. RESULTS:A supramolecular AIEgen-porphyrin co-assembly is synthesized by assembling a negatively charged AIEgen (TPE) and meso-tetra(4-carboxyphenyl)porphine (TCPP) on the surface of a cationic polymer saying poly(diallyldimethylammonium chloride) (PDDA). With the co-assembly strategy, the aggregation-induced emission of TPE, the aggregation-caused quenching characteristic of TCPP, and especially, the Förster Resonance Energy Transfer efficiency from TPE to TCPP can be manipulated by a dynamic dye assembly approach. A sensor array is thus readily constructed by varying the loading ratio of TPE and TCPP on the surface of PDDA. SIGNIFICANCE:The sensor array is determined to be powerful for pattern recognition of various glycosaminoglycans even in 100% serum media through competitive compaction or removal of the aggregated TPE and TCPP dyes on the PDDA surface. It can be successfully applied for accurate and reliable detecting trace GAG analogues in Hep samples and identifying unknown GAG samples with an accuracy of 96.88%.

STUDY QUESTION:Are higher ambient temperatures before oocyte retrieval associated with adverse oocyte-related outcomes in women undergoing their first IVF or ICSI cycle? SUMMARY ANSWER:Higher pre-retrieval temperatures are associated with reduced oocyte yield, an effect partially mediated through LH and partially counteracted by compensatory increases in FSH and estradiol (E2). WHAT IS KNOWN ALREADY:Evidence on the impact of ambient temperature on IVF/ICSI outcomes is emerging but limited. A recent study reported weak negative associations between higher ambient temperatures during folliculogenesis and oocyte yield, though the effect was modest and findings across populations remain inconsistent. However, these findings are further limited by small sample sizes, specific clinical subpopulations or geographic regions, which prevents the establishment of a generalizable dose-response relationship and leaves the underlying biological mechanisms largely unexplored. STUDY DESIGN, SIZE, DURATION:A retrospective cohort study of 58 468 women undergoing their first IVF/ICSI cycle with oocyte retrieval between 1 January 2018 and 31 May 2024. PARTICIPANTS/MATERIALS, SETTING, METHODS:Women undergoing their first IVF/ICSI cycle were included. The primary outcomes were total oocytes retrieved, mature oocytes retrieved, and 2PN (two pronuclei) fertilized oocytes. Secondary outcomes included rates of oocyte maturation, normal fertilization, and blastocyst formation. Basal sex hormone levels were measured during the early follicular phase (cycle days 2-4). Environmental exposure data, including both ambient temperature and apparent temperature (integrating air temperature and humidity), were assessed over biologically relevant exposure windows. Generalized linear mixed-effects models were used to estimate associations between temperature exposures and oocyte-related outcomes. Restricted cubic spline models assessed nonlinearity. A counterfactual causal mediation analysis examined the roles of basal sex hormones. Stratified analyses by age, BMI, antral follicle count, anti-Müllerian hormone, season, COVID-19 period, and the distance to ART center, along with sensitivity analyses, were performed to evaluate robustness. MAIN RESULTS AND THE ROLE OF CHANCE:Each 1°C increase in apparent temperature was associated with significant decreases in total oocytes retrieved (-0.20%; 95% CI: -0.28%, -0.12%), mature oocytes retrieved (-0.17%; 95% CI: -0.26%, -0.09%), and 2PN fertilized oocytes (-0.19%; 95% CI: -0.28%, -0.09%); consistent associations were observed for ambient temperature (-0.25%, -0.22%, and -0.22% per 1°C, respectively). For rate-based outcomes, no significant associations were observed for the normal fertilization rate or blastocyst formation rate; the mature oocyte rate showed small positive associations in longer-term exposure windows (3-week and 90-day). Mediation analyses indicated that LH partially mediated these associations, accounting for up to 5.23% of the total effect, while both FSH and E2 exhibited suppression effects. LIMITATIONS, REASONS FOR CAUTION:The observational design cannot definitively establish causality. Unmeasured confounding (e.g. individual indoor climate control, occupational heat exposure) and residential address-based exposure assessment are potential sources of bias. Restricting the primary analysis to completed cycles may introduce selection bias, although sensitivity analyses incorporating canceled cycles yielded consistent estimates. WIDER IMPLICATIONS OF THE FINDINGS:Higher ambient temperatures represent a modifiable environmental risk factor for oocyte yield in ART treatment, with basal sex hormone pathways, including LH as a partial mediator and compensatory shifts in FSH and E2, providing a plausible biological basis for this effect. In the context of rising global temperatures, these findings suggest that ambient thermal exposure during the peri-folliculogenesis period warrants clinical attention, and that monitoring basal sex hormone levels may help identify patients at risk of a suboptimal ovarian response in high-temperature settings. FUNDING:This study was supported in part by the Technology Innovation and Research & Development Project of the Chengdu Science and Technology Bureau (No. 2024-YF05-02159-SN), the Open Fund of Chongqing Maternal and Child Disease Control and Public Health Research Center (No. CQFYSJ01001), the Program of Inheritance and Innovation of Traditional Chinese Medicine in Chongqing (Chongqing Traditional Chinese Medicine [2022] No. 33), and Chongqing Yuzhong District Natural Science Foundation Project (No. 20240118). DISCLOSURES:All authors declare that they have no conflicts of interest related to this study. TRIAL REGISTRATION NUMBER:N/A.

Background:Neonicotinoids (NEOs) are widely used pesticides with potential endocrine-disrupting properties, yet their distribution in human reproductive matrices and associations with male reproductive hormones remain unclear. Methods:We analyzed 146 paired semen and urine samples to characterize the distribution of 14 NEOs and their metabolites (m-NEOs) and to examine associations with male reproductive hormones. Concentrations were compared across matrices, and associations with follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone (T), prolactin (PRL), estradiol (E2), and the T/LH ratio were evaluated using exposome-wide association (EWAS), deletion-substitution-addition (DSA), and mixture models. Results:Detection frequencies ranged from 43.2% to 100% in urine and 19.9% to 99.3% in semen, with Desnitro-dinotefuran (DN) and Desmethyl-acetamiprid (DM-ACE) predominant in urine and DN and Acetamiprid (ACE) enriched in semen. Partitioning analysis revealed three distinct compound clusters: (1) DN, ACE, and Imidacloprid-urea (IMI-urea) enriched in semen; (2) Dinotefuran-urea (UF), Imidacloprid (IMI), Imidaclothiz (IMIT), and 5-Hydroxy-imidacloprid (5-OH-IMI); and (3) DM-ACE, Desnitro-imidacloprid (DN-IMI), Desnitro-imidacloprid-olefin (DN-IMI-olefin), and Clothianidin (CLO) predominantly excreted in urine. In EWAS and DSA, several nominal associations with hormones were observed. Nominal associations were observed between seminal CLO and FSH, DN and E2, and urinary UF and LH, Thiacloprid (THCP) exposure showed nominal positive associations with PRL and E2. However, none remained significant after correction for multiple testing. Mixture models indicated no overall significant associations. Sensitivity analyses supported the robustness of key compound-hormone associations. Conclusion:NEOs and m-NEOs show distinct partitioning patterns between urine and semen, suggesting differential tissue distribution. Although several nominal associations with reproductive hormones were identified, none remained statistically significant after multiple-testing correction, indicating that these findings should be considered hypothesis-generating rather than conclusive. Further studies with larger populations and repeated measurements are needed to validate these associations and clarify the reproductive implications of NEO exposure in men.

To investigate how copper deficiency during pregnancy affects placental structure, metabolism, and trophoblast function, contributing to fetal growth restriction (FGR). Pregnant C57BL/6N mice were treated with ammonium tetrathiomolybdate to induce copper deficiency, with two different dosages (30 and 60 mg·kg-1·d-1) administered daily from gestational day 1 to day 14. On day 15, assessments were made on fetal growth, placental development, and spatial metabolomics. In parallel, trophoblast cells (HTR8/SVneo) were subjected to copper chelation or SLC31A1 knockdown to model copper deficiency in vitro. Cell invasiveness and proliferation were evaluated using appropriate assays, along with the measurement of molecular markers to assess the impact of copper deficiency. Copper deficiency significantly reduced maternal serum copper levels, leading to FGR, as evidenced by shorter crown-rump lengths, lower fetal weights, and altered fetal-to-placental weight ratios. Structural abnormalities in the placental junctional zone, including reduced size and altered morphology, were observed. Metabolomic analysis revealed disrupted lipid metabolism, with alterations in glycerophospholipids and fatty acids, and lipid droplet accumulation. Copper deficiency impaired trophoblast migration and invasion, linked to decreased MMP2 and MMP9 expression in vivo and in vitro. In vitro studies also showed altered lipid metabolism in SLC31A1-knockdown trophoblast cells. Copper deficiency disrupts placental structure and lipid metabolism, impairs trophoblast function, and contributes to FGR, highlighting the critical role of copper in fetal development and maternal health.

Prenatal exposure to environmental chemicals poses risks to fetal growth, yet most epidemiological studies have examined single chemical classes in isolation, leaving the broader multi-class maternal-fetal exposure landscape inadequately characterized. In addition, fetal exposure is typically inferred from cord blood concentrations, which conflates maternal body burden with transplacental transfer (TPT), obscuring the independent contribution of placental transport. We conducted a prospective cohort study screening 300 xenobiotics spanning multiple chemical classes, including PFAS, phthalate metabolites, environmental phenols/parabens, and pesticide- or industrial-related chemicals, across maternal serum and paired cord blood to define the core maternal-fetal exposome. Nineteen priority pollutants were detected at frequencies of ≥ 70% across all three biological matrices, including maternal serum, larger-twin cord blood, and smaller-twin cord blood, with PFAS representing the predominant chemical class. TPT varied substantially across compounds and differed nominally between co-twins for MBP. In linear mixed-effects models, higher TPT indices of MMP (β =  - 15.14; 95% CI: -27.66 to - 2.62), MBP (β =  - 11.61; 95% CI: -22.17 to - 1.05), and PFTrDA (β =  - 34.60; 95% CI: -67.91 to - 1.30) were nominally associated with lower birth weight. For PFTrDA, these associations were detected only when exposure was characterized by TPT, whereas corresponding cord blood concentrations were not significantly associated with fetal growth outcomes. These findings characterize a multi-class maternal-fetal exposome in twin pregnancies and provide exploratory evidence that TPT may constitute a complementary dimension of prenatal chemical exposure relevant to fetal growth.

ABSTRACT Intrahepatic cholestasis of pregnancy (ICP) is a pregnancy‐specific liver disorder characterized by elevated maternal bile acids and an increased risk of adverse fetal outcomes. Although placental dysfunction is a key contributor to ICP pathogenesis, the underlying mechanisms remain incompletely understood. Here, we combined a Sprague–Dawley rat model of ICP with spatial metabolomics to delineate region‐specific metabolic alterations within the placenta. Multivariate analysis revealed distinct metabolic signatures between ICP and control placentas, with pronounced reprogramming in trophoblast‐enriched regions. Differential metabolite and pathway analyses identified significant perturbations in glycerophospholipid metabolism, pyruvate metabolism, phospholipase D signaling, and the tricarboxylic acid cycle. Notably, phosphatidic acid (PA) was broadly elevated, whereas its downstream product phosphatidylinositol 4,5‐bisphosphate (PIP 2 ) exhibited spatially restricted accumulation in trophoblasts. Mechanistically, taurocholic acid exposure in HTR‐8/SVneo cells induced upregulation of PIP5K1, leading to PIP 2 accumulation and increased trophoblast apoptosis via disruption of the Bcl‐2/Bax balance. Silencing of PIP5K1 restored PIP 2 homeostasis and attenuated apoptosis. Importantly, these molecular alterations were recapitulated in placental tissues from ICP patients, confirming activation of the PIP5K1–PIP2 axis in vivo. Collectively, our findings identify a spatially resolved PA–PIP5K1–PIP 2 lipid signaling cascade that links bile acid–induced metabolic stress to trophoblast apoptosis and placental dysfunction in ICP.

Endometrial decidualization is essential for successful embryo implantation and pregnancy establishment. This study investigates the role of high mobility group box 1 (HMGB1) in endometrial stromal cells (hESCs) during decidualization. We found that HMGB1 expression was upregulated during decidualization in vitro and in normal decidual tissues but was reduced in decidua from patients with recurrent spontaneous abortion (RSA). HMGB1 Knockdown in hESCs impaired decidualization by disrupting stromal cell differentiation, reducing the expression of prolactin (PRL) and insulin like growth factor binding protein 1 (IGFBP1), and promoting aberrant cell proliferation. HMGB1 deficiency resulted in mitochondrial dysfunction, characterized by impaired oxidative phosphorylation, reduced ATP production, and altered mitochondrial morphology and membrane potential. Mechanistically, HMGB1 regulated the expression of PPARG coactivator 1 alpha (PPARGC1A), a key regulator of mitochondrial biogenesis and oxidative metabolism, suggesting that HMGB1 contributes to decidualization through PPARGC1A mediated metabolic regulation. Furthermore, reduced PPARGC1A expression in RSA decidua further supports the involvement of the HMGB1-PPARGC1A axis in decidual dysfunction. In addition, HMGB1 exhibited dynamic expression patterns in the uterus of early pregnant mice, with enriched expression in decidual regions at implantation sites. Collectively, these findings identify HMGB1 as an important regulator of decidualization by coordinating mitochondrial function and cellular energy metabolism, providing new insights into the molecular mechanisms underlying decidual dysfunction and pregnancy complications.

BACKGROUND:Per- and polyfluoroalkyl substances (PFAS) are persistent endocrine disruptors affecting ovarian steroidogenesis. However,their impact onovarian hyperstimulation syndrome (OHSS), a potentially life-threatening iatrogenic complication of assisted reproductive technology (ART),remains unknown. METHODS:We prospectively enrolled 374 women undergoing their first ART cycle in China (2021-2023), of whom 103 developed OHSS. We quantified 43 serum PFAS using UPLC-MS/MS,retaining 12 congeners with detection frequencies > 50%for subsequent analyses, and four congeners with detection frequencies > 70% were included in mixture analyses. Associations were evaluated using modified Poisson regression, restricted cubic splines, Bayesian kernel machine regression (BKMR), and quantile g-computation (qgcomp). RESULTS:In continuous variable models, log10-transformed concentrations of PFNA, PFDA, and PFUdA were significantly associated with an elevated risk of OHSS (adjusted RR [aRR] = 1.30, 95% CI: 1.08-1.58, p = 0.006; aRR = 1.21, 95% CI: 1.01-1.44, p = 0.034; and aRR = 1.42, 95% CI: 1.16-1.73, p < 0.001, respectively). Tertile analyses further confirmed these dose-dependent positive associations. Mixture analyses via BKMR identified PFNA and OBS as dominant risk contributors, with a non-monotonic overall mixture-response relationship. These adverse associations were primarily concentrated in women < 35 years. CONCLUSIONS:To our knowledge, this study provides the first epidemiological evidence linking specific PFAS congeners and their mixtures to elevated OHSS risk during ART, supporting the rationale for preconception PFAS biomonitoring and targeted exposure reduction in reproductive-age women.

Background: It is meaningful but challenging to develop convenient and accurate analytical methods for heparin sensing, owing to its complex and heterogeneous chemical structure. Although indicator displacement assay offers a promising alternative, the number of suitable host molecules capable of accommodating heparin remains limited. Results: Two positively charged porphyrin-based polymers, A30 and A32, are synthesized via alkylation reactions between 5,10,15,20-tetra(4-pyridyl)porphyrin and corresponding linkers of 1,2-diiodoethane and p-Xylylene dibromide, respectively. These polymers are designed to interact effectively with heparin through multiple hydrogen bonding and electrostatic interactions. Especially, A30 and A32 exhibit graphene oxide-like fluorescence quenching behavior, enabling their use as a general platform for constructing non-fluorescent polymer-dye ensembles. Therefore, a four-component sensor array named PSA is constructed by simple mixing A30 and A32 with representative dyes for heparin sensing. Significance: The PSA sensor array demonstrates satisfactory performance in discriminating common glycosaminoglycans, including heparin, chondroitin sulfate, and hyaluronic acid. Furthermore, it has been successfully applied for quantifying heparin concentration in diluted serum samples, detecting glycosaminoglycan contaminants in heparin, and accurately identifying unknown glycosaminoglycan samples in an accurate and convenient way.

BackgroundThere is currently no consensus on the etiology, pathogenesis, or treatment of endometriosis (EM). The discovery of disease-associated plasma proteins with causal genetic evidence provides an opportunity to identify new EM biomarkers and therapeutic targets.MethodsProtein quantitative trait loci (pQTLs) were derived from plasma proteomic associations in the UK Biobank Pharma Proteomics Project (UKB-PPP). Genetic associations with EM were obtained from the FinnGen cohort. The associations between proteins and the risk of EM were estimated by cis-Mendelian randomization (cis-MR) and validated using the GWAS catalog dataset of EM. Colocalization, protein-protein interaction (PPI) analysis, functional enrichment analysis, transcriptome differential expression gene (DEG) analysis and druggability evaluation were further performed to explore potential biomarkers and therapeutic targets for EM.ResultsOverall, genetically predicted levels of 23 plasma proteins were associated with EM risk, with five proteins validated via replication analysis (ALPI, KHK, HSPG2, STXBP1, and POLR2F). Lower levels of genetically predicted ALPI (odds ratio [OR]: 0.89, 95% confidence interval [CI] 0.83-0.95), HSPG2 (OR: 0.81, 95% CI 0.75-0.88), POLR2F (OR: 0.51, 95% CI 0.36-0.73), and STXBP1 (OR: 0.75, 95% CI 0.64-0.86) were associated with an increased risk of EM. Elevated levels of KHK (OR: 1.09, 95% CI 1.05-1.13) were associated with an increased risk of EM. We also identified ALPI, KHK, HSPG2, STXBP1, and POLR2F as potential drug targets and biomarkers for EM.ConclusionA series of comprehensive analyses emphasized the potential role of ALPI, KHK, HSPG2, STXBP1, and POLR2F in EM and suggested that these genes could be developed into exact biomarkers and therapeutic targets for this condition in future research.