Processing-bodies (PBs) are cytoplasmic membraneless condensates essential for RNA regulation. They share structural and functional similarities with germ granules and are critical for primordial folliculogenesis, a developmental process highly susceptible to exogenous insults. Aristolochic acid I (AAⅠ), a pervasive environmental toxin from Aristolochiaceae plants, is known to cross the placental barrier and induce fetal ovarian oxidative stress. Here, using a mouse model of maternal AAⅠ exposure (16.5 to 18.5 days post-coitus), we demonstrate that this prenatal insult causes profound defects in offspring folliculogenesis. AAⅠ induces PB enlargement and protein accumulation, causing these condensates to acquire germ granule-like properties. Specifically, AAⅠ upregulates the germ granule component NANOS3 to drive condensate enlargement. Multi-omic profiling further revealed that these aberrant condensates recruit germ granule-associated proteins and exhibit altered RNA profiles. Concurrently, neonatal oocytes exhibited enlarged Balbiani body-like structures alongside intermitochondrial cement, a feature not previously reported in oocytes. Furthermore, AAⅠ promotes the recruitment of N6-methyladenosine (m6A)-related factors into PBs, accompanied by elevated global m6A levels and the upregulation of YTHDF2. We show that YTHDF2 is essential for recruiting NANOS3 to PBs; truncating the intrinsically disordered regions (IDRs) or RNA-binding domains of either protein impairs their interaction. These perinatal perturbations manifest in adulthood as primary ovarian insufficiency (POI)-like dysfunction, including reduced ovarian reserve, compromised oocyte quality, and hormonal deficits. Overall, our work uncovers a NANOS3-YTHDF2 axis that drives germ granule-like PB adaptations, illustrating how fetal environmental insults disrupt folliculogenesis and serve as an etiological driver for adult-onset POI. Female reproductive health depends on the establishment of the ovarian follicle pool before birth, a delicate process highly vulnerable to environmental toxins. This study investigates how Aristolochic acid I (AAⅠ), a widespread, plant-derived toxin that accumulates in the food chain, disrupts folliculogenesis in mice. We found that maternal AAⅠ exposure delayed primordial folliculogenesis in neonatal mouse ovaries. Specifically, AAⅠ abnormally upregulates germline factor NANOS3 and drives the enlargement of processing-bodies (PBs), which regulate RNA metabolism. AAⅠ causes these PBs to recruit germ granule-associated proteins and alters the profiles of PB-associated RNAs. Furthermore, AAⅠ elevates m6A RNA modifications and upregulates the m6A reader YTHDF2. Our cellular assays reveal that YTHDF2 is essential for recruiting NANOS3 into PBs. Most importantly, this neonatal ovarian damage has long-lasting consequences. Female offspring maternally exposed to AAⅠ develop primary ovarian insufficiency (POI)-like dysfunction in early adulthood, characterized by a severely reduced ovarian reserve, poor oocyte quality, and hormonal deficits. Ultimately, our findings illustrate how environmental insults during fetal development can impair female fertility and trigger adult-onset ovarian dysfunction.

Maternal pre-pregnancy obesity is a well-established risk factor for a spectrum of adverse pregnancy outcomes, yet the molecular mechanisms through which obesity disrupts the early endometrial microenvironment have remained incompletely understood. Although placental abnormalities in obesity are well-documented, the developmental origins of these defects—particularly the molecular events during embryo implantation and decidualization that establish the foundation for normal placentation—have received limited attention. Here, integrating UK Biobank data with mechanistic studies in experimental models, we demonstrate that high-fat diet-induced pre-pregnancy obesity and high-fat exposure disrupt decidual angiogenesis by impairing stromal–endothelial communication. Specifically, high-fat exposure suppresses Hnrnpa2b1 binding to the mPGES-1 promoter, inhibiting prostaglandin E2 (PGE2) synthesis and subsequent vascular endothelial growth factor A secretion from stromal cells and leading to defective vascularization. Functional rescue experiments, including 3D biomimetic chip co-culture systems and targeted in vivo overexpression, confirm that restoring the Hnrnpa2b1/mPGES-1/PGE2 axis reinstates angiogenic competence and improves pregnancy outcomes. Our findings reveal a previously unrecognized metabolic-transcriptional cascade linking high-fat diet-induced obesity to endometrial vascular fragility and propose new diagnostic and therapeutic strategies for obesity-related reproductive failure.

This study aimed to explore the association between leukocyte telomere length (LTL) and premature rupture of membranes (PROM). This cross-sectional study included a total of 170,841 participants from the UK Biobank. Restricted cubic spline model (RCS) and logistic regression were used to evaluate the relationship between LTL and PROM. Subgroup analysis and interaction tests were conducted to test the stability of the results. RCS model showed a notable non-linear association between LTL and PROM. The logistic regression model further revealed that longer LTL was associated with an increased odds of developing PROM after fully adjusting for confounding factors. For every 1 unit increase in the original LTL, the odds of PROM significantly increases by 1.97 times (OR = 1.97, 95% CI:1.30-2.95). Compared with the Q1 of LTL, the PROM odds of Q3 and Q4 increased by 33% (OR = 1.33, 95% CI: 1.11-1.59) and 31% (OR = 1.31, 95% CI: 1.09-1.56). Finally, subgroup analysis found that age and education level play an important role in regulating the relationship between LTL and the odds of PROM. The research results indicate a non-linear relationship between LTL and PROM, and longer LTL is associated with higher PROM odds, which is influenced by age and education level.

Background: Neonatal respiratory failure (NRF) is the most common issue among premature and full-term infants admitted to the neonatal intensive care unit. The incidence rate and morbidity of NRF in clinical practice, especially in twin pregnancy, remain high. Methods: A total of 3,721 women with twin pregnancies were included in this retrospective study. The Lasso regression was employed to optimize the selection of relevant features. Following this, multivariate logistic regression analysis was utilized to construct a nomogram. The predictive performance of models was evaluated using the receiver operating characteristic (ROC) curve, calibration plot, and clinical decision curve. Furthermore, nine machine learning models were constructed for predicting NRF. Accuracy, precision, recall, F1 score, and ROC curve were used to evaluate the predictability of machine learning models. Results: The results of multivariate logistic regression analysis revealed that gestational age (GA), heparin, monochorionic monoamniotic (MCMA), placenta accreta, and placenta previa emerged as distinctive independent risk factors for NRF. Subsequently, a nomogram was constructed, incorporating these independent prognostic factors. In the training set, the nomogram exhibited the area under the curve (AUC) value of 0.908, while in the validation set, this metric remained high at 0.899. Among the machine learning models, long short-term memory (LSTM) and ensemble learning (EL) outshone the others, demonstrating the best performance with an AUC value of 0.91 in the validation set. The nomogram and machine learning models employed in this study demonstrated a robust and reliable predictive performance. Conclusions: The nomogram and machine learning models developed in this study prove to be effective and userfriendly tools for predicting the likelihood of NRF.

Extreme temperature events have become more frequent due to industrialization, urbanization, and climate change, leading to health risks. Women with twin pregnancies are particularly vulnerable, yet research on how these temperatures affect gestational diabetes mellitus (GDM) remains limited. This study investigates the impact of extreme temperatures on GDM incidence in twin pregnancies, focusing on identifying critical periods of susceptibility. Data from 3,769 twin pregnancies were analyzed using temperature information from local meteorological stations. Extreme temperatures, defined as heat waves and cold spells, were categorized by daily maximum, minimum, and mean temperatures. The relationship between the frequency and duration of these events and GDM risk was evaluated through multivariable logistic regression, while a distributed lag nonlinear logistic model assessed temperature impacts across gestational weeks. Results from 1,063 GDM cases revealed significant differences in incidence across screening temperatures. GDM risk increased with days exceeding the 90th percentile temperature in the four weeks prior to screening, as well as with the frequency of cold spells and heatwaves earlier in pregnancy. Both high and low extreme temperatures during specific gestational weeks were associated with higher GDM risk, suggesting these factors should be considered in preventive strategies for twin pregnancies.

Maintaining normal thyroid function is crucial in pregnancy, and the thyroid hormone signaling pathway is involved in embryo implantation. However, the regulation of iodothyronine deiodinase 2 (DIO2), which is the central hub controlling thyroid hormone signaling, and the intracellular pathway activated by triiodothyronine (T3) binding to the thyroid hormone receptor (THR) in endometrial cells, remains unclear. Here, we demonstrate that DIO2 expression increases in endometrium during the establishment of endometrial receptivity and is involved in this process. Iopanoic acid inhibition of DIO2 in vivo can cause a delayed receptive state. In vitro adhesion models have consistently confirmed that knocking down DIO2 in epithelial cells inhibited receptivity establishment. Membrane lipidomics was performed to explore how DIO2 regulates the morphological transformation of endometrial epithelial cells. We found that the deletion of Dio2 inhibited the increase in the degree of lipid unsaturation, which subsequently decreased membrane fluidity. Transcriptomics analysis was employed to explore the downstream target gene of T3-THR signaling mediated by Dio2-mediated T3-THR signaling, and Scd1 is confirmed as the direct target gene of THR in endometrial epithelial cells. These data reveal that DIO2 could regulate lipid metabolism by targeting Scd1 through the T3-THR signaling pathway, thereby modifying membrane fluidity of endometrial epithelial cells and promoting cell morphological transformation to establish endometrial receptivity. These findings contribute to filling the gap in downstream pathways activated by T3-THR signaling in endometrial cells and provide insights into the new therapeutics, prediagnosis, and preventive strategies for the derailment of endometrial receptivity and subsequently adverse "ripple effect" including infertility.

BACKGROUND:Accelerated industrialization globally has intensified air pollution, but the susceptibility periods for extreme air pollution in twin pregnancies remain undefined. METHODS:This study investigated the association between extreme air pollution exposure and preterm birth risk in twin pregnancies. Data on 3623 twin pregnancies in Chongqing from 2017 to 2022 and air pollution readings from 12 monitoring stations were analyzed using distributed lag non-linear quasi-Poisson regression models. Additionally, four extreme air pollution indices were developed to assess the cumulative effects of lagged exposures on preterm birth risk through multivariate logistic regression. RESULTS:Compared to the lower quartile, the 95th percentile of extreme air pollution exposure showed a positive correlation between concentrations of PM2.5, PM10, NO2, SO2 and CO and preterm birth risk in twin pregnancies, with O3 inversely correlated. Sensitive periods for air pollutants were different. 8-12 and 27-35 gestational weeks were identified for PM2.5; 6-13 and 27-35 gestational weeks were identified for PM10; 5-14 and 21-33 gestational weeks were identified for NO2; 4-15 and 24-36 gestational weeks were identified for SO2; 4-11 and 29-33 gestational weeks were identified for CO. PM2.5, PM10, SO2 and O3 showed cumulative effects across short and long lags, while CO showed a long-term effect. Notably, NO2 exhibited a protective effect during all lag periods. CONCLUSION:The study highlights gestational windows of 8-11 and 29-33 weeks as highly sensitive to extreme pollution for preterm birth in twin pregnancies, with marked risk increases during 0-3, 0-6 and 0-9-month lag periods.

Alzheimer's disease (AD) is a highly inheritable neurodegenerative disorder for which pathway-specific genetic profiling provides insights into its key biological mechanisms and potential treatment targets. Traditional disease-pathway analyses for AD have certain limitations, such as environmental interference and arbitrary sample division. We present a comprehensive framework that starts with genome data, avoiding these drawbacks and offering intrinsic pathway-specific genetic profiling for AD. Whole genome sequencing data from 173 individuals were used to quantify transcriptomes in 14 brain regions, estimate individual-level pathway variant scores, and analyze AD risk for each patient. These results were combined to identify AD-related pathways and quantify their interactions. The predicted expression levels were consistent with previous findings, and the estimated AD risk showed a significant correlation with Braak/Thal scores. A total of 3798 pathways were identified as potentially associated with AD, with about 19.7 % previously reported. The pathways identified as AD risk related primarily address six core biological themes, including: Immunity and inflammation, Metabolism, Protein homeostasis, DNA/RNA and Epigenetics, Synapse and structure, Cell cycle. Specifically, key pathways, such as NF-κB signaling and GSK3β activation, were linked to AD pathogenesis. The interactions among pathways highlighted shared gene functions in AD. In summary, we provided an effective framework for disease-pathway analysis, revealing the interdependence or compensatory effects of pathways in AD.

Triple-negative breast cancer (TNBC) remains a formidable clinical challenge owing to its aggressive behavior, immunosuppressive tumor microenvironment, and lack of effective targeted therapies. To address these limitations, we developed a magneto-photo-acoustic responsive nanoplatform (MnFe2O4-erastin-perfluoropentane nanoparticles [MEPNPs]). This nanoplatform features 3-tiered therapeutic innovations: (a) Multimodal imaging-guided precision therapy: The superparamagnetic property of MnFe2O4 enabled magnetic resonance and photoacoustic imaging, allowing real-time visualization of tumor margins. (b) Spatiotemporally controlled ferroptosis activation: Magnetic targeting enhanced the tumor accumulation of MEPNPs, while near-infrared irradiation triggered perfluoropentane vaporization for burst erastin release. This dual strategy combinationally suppressed glutathione peroxidase 4 and amplified the accumulation of lipid peroxides, achieving the amplification of ferroptosis. (c) Immunogenic tumor microenvironment reprogramming: MEPNP-induced immunogenic cell death promoted dendritic cell maturation and CD8+ T-cell infiltration, effectively converting immunologically "cold" TNBC tumors into "hot" phenotypes. In TNBC models, MEPNP treatment elicited remarkable therapeutic outcomes: primary tumor suppression, reduction in lung metastasis, and an extended median survival period exceeding 45 d. The transcriptome sequencing results showed that there were 6,198 differentially expressed genes in the treatment group. These included the up-regulation of ferroptosis drivers such as SLC39A14, as well as the down-regulation of antioxidant regulators such as SLC7A11 and SLC3A2. Additionally, Kyoto Encyclopedia of Genes and Genomes pathway analysis confirmed that the "ferroptosis" and "T-cell differentiation" pathways were specifically activated. This work establishes a novel "theranostic-immunomodulatory" paradigm that integrates magnetic targeting, ferroptosis potentiation, and immunogenic-cell-death-mediated immune memory. By orchestrating physical energy conversion, MEPNPs provide a spatially focused and immunologically amplified strategy to overcome TNBC therapeutic resistance.

Background:This study compares the prevalence of placenta accreta in singleton and twin pregnancies and examines its impact on adverse perinatal outcomes, exploring whether twin gestation increases the risk of poor outcomes in placenta accreta cases. Methods:A multivariate logistic regression analysis assessed the link between twin pregnancy and placenta accreta, comparing associated adverse perinatal outcomes in twin vs. singleton pregnancies. Stratified and interaction analyses explored clinical characteristics' relationship with placenta accreta. The Restrictive Cubic Spline (RCS) model evaluated the impact of placenta accreta on caesarean section and postpartum haemorrhage at different gestational ages. A comparative analysis examined clinical features and perinatal outcomes between twin and singleton pregnancies with placenta accreta. Finally, mediation analysis was used to determine if placenta accreta mediates the effect of twin gestation on caesarean section and postpartum haemorrhage. Results:In a large cohort study of 16 908 pregnancies, including both twin and singleton pregnancies, conducted in Chongqing, China, the risk of placenta accreta increased by 51% in twin gestations, with haemorrhagic placenta accreta rising by 133%. This condition significantly heightened the risk of adverse perinatal outcomes in both singleton and twin pregnancies, with twin pregnancies exhibiting higher risks. In twins, the risk of preterm birth was 1.77 (95% confidence interval (CI) = 1.24, 2.52), caesarean section was 4.87 (95% CI = 3.00, 7.90), postpartum haemorrhage was 3.73 (95% CI = 1.95, 7.13), and uterine rupture was 26.42 (95% CI = 2.28, 306.63). Additionally, placenta accreta showed different interactions with various factors in both twin and singleton pregnancies, influencing distinct outcomes. Restricted Cubic Splines (RCS) model analysis indicated an increasing trend in the risk of caesarean section and postpartum haemorrhage associated with placenta accreta across all gestational ages in both singleton and twin gestations. In patients with placenta accreta, the risks of preterm birth, caesarean section, pelvic inflammatory disease, atonic postpartum haemorrhage, and premature rupture of membranes in twin gestations were 6.77, 2.39, 2.54, 5.84, and 2.93 times higher, respectively, than in singleton gestations. Finally, mediation causal analysis revealed that the effect of twin gestation on caesarean section included both a direct effect and an indirect effect mediated through placenta accreta. For postpartum haemorrhage, the effect of twin gestation was mediated through placenta accreta. Conclusions:Twin gestation, regardless of known risk factors, increases the risk of placenta accreta and adverse perinatal outcomes compared to singleton pregnancies. Antenatal interventions and delivery risk management are essential for twin pregnancies with placenta accreta.

Successful placental development and pregnancy rely on effective extravillous trophoblast (EVT) invasion. The mechanisms underlying inadequate EVT invasion in recurrent spontaneous abortion (RSA) remain unclear. WAS/WASL interacting protein family member 1 (WIPF1), the key regulator of cytoskeletal dynamics, is exclusively expressed in first-trimester placental EVTs. Knockdown experiments revealed WIPF1's crucial involvement in successful placental development; reduced levels impaired cell migration, while overexpression induced the opposite effects. Moreover, WIPF1 knockdown in hTSC-derived EVTs hampered trophoblast differentiation. WIPF1 interacted with ACTN4 to regulate podosome formation, matrix degradation, and actin polymerization, potentially mediated by its ARG54 site. Notably, WIPF1 was significantly down-regulated in human RSA patient EVTs and RSA mice trophoblast giant cells (CBA/J × DBA/2). This association suggests WIPF1 as a potential key player in RSA pathogenesis. In conclusion, our study spotlights WIPF1 as a pivotal factor in EVT invasion, emphasizing its multifaceted roles and implications in pregnancy complications like RSA.

The extensive utilization of plastics has heightened concerns regarding microplastics exposure. However, the effects of polystyrene microplastics (PS-MPs) on early pregnancy remain inadequately investigated. This study aimed to examine the impact of PS-MPs on decidualization and embryo implantation in female mice, as well as the reproductive function of their offspring following maternal exposure to PS-MPs. We investigated the harmful effects of different PS-MPs sizes on mouse endometrial stromal cells (mESCs) during in vitro decidualization. Pregnant mice were orally given various concentrations of PS-MPs to examine their impact on decidualization. We evaluated oxidative stress and inflammation markers to understand their roles in abnormal decidualization. Additionally, we assessed potential reproductive health impacts on female offspring. Our findings indicated that 5 µm PS-MPs effectively penetrated mESCs and significantly disrupted decidualization compared to smaller or larger particles. Pregnant mice that were exposed to 5 µm PS-MPs at a dose of 1000 mg/(kg·day) exhibited substantial reductions in the decidual area and downregulation of decidualization markers such as BMP2. Inflammatory cytokines increased significantly in mESCs following 5 µm PS-MPs exposure, and the elevated malondialdehyde levels in uterine tissue were mitigated by antioxidant treatment. Moreover, offspring exhibited decreased uterine wet weight, uterine organ coefficients, decidual areas, and expression of BMP2 due to maternal exposure to PS-MPs. These results highlighted the detrimental effects of PS-MPs on maternal decidualization and embryo implantation, suggesting a link to oxidative stress and inflammation, and maternal exposure to PS-MPs during pregnancy impaired reproductive function in offspring females.

The receptive endometrium is a prerequisite for successful embryo implantation, and abnormal endometrial receptivity would lead to infertility. Many key proteins involved in endometrial receptivity have been confirmed to undergo post transcriptional modifications. However, there are limited reports on deubiquitination modification during this process. Our previous studies found that Rictor participated in the endometrial receptivity, and maintained at a high level in the endometrium during implantation, but the mechanism for maintaining stability of Rictor protein remains unclear. Here, we showed that USP9X expression in endometrium was dynamic with the establishment of endometrial receptivity, and promoted the protein stability of Rictor through deubiquitination. Inhibition of USP9X could suppress the adhesion action of trophoblast cells to endometrial epithelial cells, reduce the filamentous pseudopodia of epithelial cells, and inhibit the epithelial mesenchymal transformation. Rictor is partially responsible for the derailment of epithelial cell transformation in response to USP9X inhibition. Membrane fluidity mediated by lipid metabolism is involved in regulation of Rictor on endometrial receptivity. This study revealed the role of USP9X in endometrial receptivity for the first time, and confirmed that Rictor was the target protein of USP9X in endometrium. In addition, we described the unique lipidomics characteristics of the endometrial epithelial cells regulated by Rictor. These data would further improve the molecular network of endometrial receptivity, supplement the regulatory factors of lipid metabolism in endometrial cells, and provide insights into the new therapeutics, pre-diagnosis and preventive strategies for the derailment of endometrial receptivity and subsequently adverse "ripple effect" including infertility.

The placenta serves as a vital interface for fetal-maternal exchange, relying on trophoblast differentiation for development. This process involves cytotrophoblasts (CTBs) transitioning into syncytiotrophoblasts (STBs) and extravillous trophoblasts (EVTs), driving placental maturation. Focal adhesion kinase (FAK), a key cytoplasmic tyrosine kinase, regulates cellular processes such as proliferation, survival, and signaling. However, its role in trophoblast differentiation and metabolism remains unclear. Here, using human trophoblast stem cells (hTSCs) and trophoblast cell lines (BeWo, HTR8/SVneo), we investigated FAK signaling in trophoblast lineage differentiation. Inhibiting the FAK signaling pathway suppresses MAPK pathway activity, reduces glycolytic metabolism and impairs trophoblast syncytialization. Additionally, blocking FAK with the inhibitor Defactinib disrupts EVTs cytoskeleton and impairs migration, invasion, and differentiation potential. Notably, reduced FAK signaling is observed in patients with recurrent spontaneous abortion (RSA), suggesting a role in RSA pathogenesis. Our findings highlight FAK as a pivotal regulator of trophoblast lineage development, linking it to placental function and RSA. This study offers new insights into placental disorders and potential therapeutic targets.

Polycystic ovary syndrome (PCOS) is a prevalent endocrine and metabolic disorder affecting women of reproductive age. Oxidative stress (OS) is suggested to play a significant role in the development of PCOS. Using antioxidants to reduce OS and maintain a healthy balance in the body could be a novel treatment approach for PCOS. This study analyzed transcriptome data from the Gene Expression Omnibus database, focusing on genes associated with OS. By implementing two machine learning algorithms, three OS-related biomarkers—HMOX1, MMP9, and KLF2—were successfully identified. To evaluate the diagnostic potential of these biomarkers, a Logistic regression model was employed. Additionally, granulosa cells were collected from healthy individuals and infertile women with PCOS, and the reliability of HMOX1, MMP9, and KLF2 was verified by quantitative real-time PCR experiments. Furthermore, small molecule drugs targeting proteins encoded by genes HMOX1 and MMP9 were predicted through the Drug Signature Database. Molecular docking of drugs to proteins identified two antioxidants, butein and demethoxycurcumin, as potential candidates for PCOS therapy.

PurposeEndometriosis (EM) and recurrent spontaneous abortion (RSA) exhibit clinical associations, yet their shared molecular mechanisms remain unclear. This study aimed to identify shared molecular mechanisms and potential hub genes underlying EM and RSA.MethodsDifferentially expressed genes (DEGs) were identified from EM (GSE7305) and RSA (GSE165004) datasets. Functional enrichment and weighted gene co-expression network analysis (WGCNA) revealed shared pathways and key modules. Venny software was used to identify hub genes between DEGs and key module genes. The diagnostic value of FXYD1 was assessed via ROC analysis. Regulatory networks and immune cell infiltration were explored. Pan-cancer analysis was conducted to assess FXYD1's expression profile across tumor types. Single-cell RNA sequencing validated FXYD1 expression in EM tissues, maternal-fetal interface and RSA samples.ResultsDEGs in EM and RSA were enriched in pathways associated with abnormal proliferation, immune dysfunction, and developmental regulation. FXYD1 was identified as a shared hub gene, upregulated in both conditions, with potential diagnostic value. It was correlated with immune cell populations, particularly natural killer (NK) cells. Pan-cancer analysis revealed widespread FXYD1 downregulation across multiple cancer types. Single-cell RNA sequencing confirmed elevated FXYD1 expression in stromal and decidual cells of RSA tissues, implicating its role in impaired decidualization.ConclusionsFXYD1 emerges as a critical molecular link between EM and RSA, potentially contributing to decidualization dysfunction. Its dysregulation may underlie the shared pathophysiology of these conditions, offering new insights into their molecular mechanisms.

Background: Lung cancer is the primary cause of cancer-related mortality, but the molecular mechanisms behind this malignancy remain unclear. Methods: The Cancer Genome Atlas (TCGA) online database and tissue chips were used to analyze the expression levels of tumor necrosis factor receptor-associated factor 2 (TRAF2)- and non-catalytic region of tyrosine kinase adaptor protein (NCK)- interacting kinase (TNIK) protein in lung cancer. A549 and PC-9 lung adenocarcinoma (LUAD) cells with stable TNIK knockdown were generated by lentivirus infection. The tumor phenotypes were subsequently examined both in vitro and in vivo. The TCGA online database and RNA-sequencing of TNIK-knockdown cells were used to study the molecular mechanism underlying the TNIK-mediated phenotype of LUAD cells. The effects of TNIK knockdown on focal adhesion dynamics and mitosis were examined by indirect immunofluorescence and Western blot, on the sensitivity to chemotherapy drugs by cell counting kit-8 (CCK-8) assay, on apoptosis by flow cytometry, and on cell proliferation by 5-ethynyl-2′-deoxyuridine (EDU). Results: TNIK was highly expressed in LUAD (p < 0.0001), predominantly in the cytosol. Phenotype assays revealed that TNIK knockdown in LUAD cells led to a significant increase in cell spreading (p < 0.0001), but also inhibition of cell growth and movement (p < 0.01). Mechanistically, TNIK was found to regulate F-actin and microtubule organization, as well as the Ras homolog gene family (RHO)/RHO-associated kinase 2 (ROCK2)/LIM motif-containing protein kinase 1 (LIMK1) signaling pathway, thereby playing a crucial role in the control of focal adhesion turnover and mitosis. Additionally, the silencing of TNIK enhanced the sensitivity of LUAD cells to chemotherapeutic drugs. Conclusions: Our findings suggest that TNIK regulates focal adhesion turnover and mitosis to promote tumor malignancy via the RHO/ROCK2/LIMK1 pathway. The combination of TNIK targeting with chemotherapeutic drugs could be an effective strategy to overcome resistance in LUAD.

Background: Recent studies have primarily focused on the impact of environmental factors on gestational diabetes mellitus (GDM) in singleton pregnancies, with limited research on their effects in twin pregnancies. This study investigates how seasonal variations and environmental exposures impact GDM incidence and its subtypes in twin pregnancies, a high-risk group. Methods: In this retrospective analysis of 3769 twin pregnancies, we categorized recruited participants into GDM and non-GDM groups. We examined the effect of the screening season on oral glucose tolerance test (OGTT) glucose values and the incidence of GDM and its subtypes. Multivariable logistic regressions adjusted for confounders assessed the impact of first and second trimester temperatures and air pollutants on GDM risk. Interaction terms evaluated the combined effects of environmental factors on GDM incidence. Results: Seasonal changes significantly influenced GDM risk, with summer presenting the highest risk (p < 0.05). The first trimester's cooler temperatures were inversely related to GDM; T mean was significantly and negatively associated with 1-h PG and AUC for glucose, with adjusted β (95% CI) of -0.009 (-0.017, -0.001) and -0.719 (-1.406, -0.031), respectively. While warmer second trimester temperatures increased the risk, T mean was positively associated with FBG, 1-h PG, 2-h PG, and AUC for glucose, with adjusted β (95% CI) of 0.003 (0.001, 0.005), 0.018 (0.009, 0.026), 0.019 (0.011, 0.027), and 1.723 (0.998, 2.448), respectively. Air pollutant exposure showed varying correlations with GDM risk, with ozone (O3) levels consistently posing a risk. Higher O3 exposure in the first and second trimesters was associated with increased odds of GDM, with OR (95% CI) of 1.057 (1.004, 1.112) and 1.052 (1.011, 1.096), respectively. Interaction analysis indicated that certain environmental conditions in the first trimester could reduce GDM risk, while others, particularly involving O3, increased it. Conclusion: Environmental temperatures and air pollutants, especially O3, are associated with GDM risk in twin pregnancies, with differing effects between trimesters. These findings suggest that environmental factors should be considered in GDM screening and prevention strategies for twin pregnancies. Further research is needed to understand the underlying mechanisms and to develop trimester-specific interventions.

Recurrent spontaneous abortion (RSA) is a pregnancy-related condition characterized by a complex etiology. While placental trophoblast dysfunction is strongly associated with the development and progression of RSA, the underlying molecular mechanisms remain poorly understood. In this study, we observed a significant decrease in the expression of MYB Proto-Oncogene Like 2 (MYBL2) in the villous tissue of patients with RSA and the placentas of abortion-prone (AP) mice. Utilizing human trophoblast stem cells (hTSCs), we identified MYBL2 as a critical regulator of hTSCs stemness maintenance, promoting the expression of the stemness-associated genes Tumor protein p63 (TP63) and TEA Domain Transcription Factor 4 (TEAD4). Furthermore, MYBL2 facilitates the differentiation of hTSCs into extravillous trophoblast (EVT) by positively regulating Ajuba LIM Protein (AJUBA) expression. Using HTR-8/SVneo cell line, an immortalized EVT-like model, we found that MYBL2 positively regulates AJUBA expression by binding to the distal region of the AJUBA promoter. Additionally, the MYBL2–AJUBA axis enhances the migration and invasion of HTR-8/SVneo cells by suppressing the Hippo signaling pathway. Our study indicates that the dysregulation of MYBL2 expression in placental trophoblasts is associated with the pathogenesis of RSA, highlighting its potential as a therapeutic target for this condition.

This cohort study assessed the stillbirth rate and neonatal comorbidities associated with the timing of delivery in twin pregnancies following in-vitro fertilization (IVF) treatments. This retrospective study encompassed 1596 twin pregnancies and categorized participants into spontaneous conception (SC) and IVF groups. The investigation initially assessed the impact of IVF on maternal and neonatal outcomes post-delivery, followed by an exploration of the prospective risk of stillbirth and incidence of stillbirth under IVF and gestational age stratification. Subsequently, multivariable Cox regression analysis was conducted to determine any significant difference in twin mortality with or without IVF. Additionally, post-delivery maternal and neonatal comorbidities rates are examined within the context of IVF and gestational age categories using multivariable logistic regression and restricted cubic splines to investigate trends in neonatal comorbidities with and without IVF. The objective was to optimize delivery timing to balance the risk of stillbirth associated with continued pregnancy against the risks of late preterm birth and neonatal complications, thereby achieving the best possible maternal and infant health outcomes. The study revealed that twin pregnancies conceived through IVF were associated with higher maternal age and pre-pregnancy body mass index (PBMI) compared to the SC group, yet there were no significant differences in the incidence of maternal and neonatal outcomes post-delivery. While the prospective risk of stillbirth and the rate of stillbirth was higher in the IVF group at each delivery time point, these differences are not statistically significant after adjusting for confounding factors in the Cox regression analysis. The incidence of post-delivery maternal and neonatal outcomes in the IVF group was not significantly different from the SC group across various delivery times and after adjustment using logistic regression and restricted cubic splines, gestational age significantly affected the risk of composite neonatal outcomes (p < 0.05). In the IVF group, compared to a median gestational age of 37 weeks, both late preterm and pregnancies delivered beyond 37 weeks showed an increasing trend in the risk of composite neonatal outcomes. Conversely, in the SC group, the risk of composite neonatal outcomes showed a decreasing trend with the extension of gestational weeks at delivery. In twin pregnancies resulting from IVF treatment, both the prospective risk of stillbirth and the rate of stillbirth were higher compared to those in the SC group. Considering the associated risks of stillbirth and neonatal complications, delivery around 37 weeks may be associated with more favorable outcomes. However, this observation does not establish 37 weeks as the definitive optimal time for delivery. The findings suggest that further research is needed to explore the best delivery timing for IVF twin pregnancies and to guide clinical decision-making for optimizing pregnancy outcomes.