BACKGROUND:Most existing evidence on secondhand smoke (SHS) exposure and lung cancer in never-smoking women comes from case-control studies, while findings from prospective cohort studies remain inconsistent. Regional variation in SHS-associated risk is poorly understood, and the potential impact of reducing SHS exposure in never-smoking women has not been quantified. METHODS:We analyzed data from 20,401 never-smoking women in the Wenling lung cancer screening cohort (Zhejiang, China), applying Cox proportional hazards regression models to evaluate the association between SHS exposure and lung cancer risk. We further conducted a meta-analysis of published studies to examine regional variation and estimated the population attributable fraction (PAF) of SHS exposure for lung cancer incidence in never-smoking women. RESULTS:SHS exposure was associated with an increased lung cancer risk (HR=1.52, 95% CI: 1.19-1.95). Lung cancer risk tended to increase with higher smoking intensity, with a linear trend observed for cigarettes per day, years of smoking, and pack-years of co-residents and colleagues (Poverall<0.05; Pnon-linear>0.05). In meta-analysis, the pooled relative risk was higher in Asia than in Europe and America [RR= 1.31 (95% CI: 1.20-1.43) vs 1.13 (95% CI: 1.04-1.23); Phet = 0.039]. The PAF suggested that 16.9% (95% CI: 11.6%-22.0%) of lung cancer cases among never-smoking women could be attributed to SHS exposure. CONCLUSIONS:SHS exposure is a risk factor for lung cancer in never-smoking women, with stronger effects observed in Asian populations. Reducing SHS exposure could prevent a substantial proportion of lung cancer cases, underscoring the urgent need for strengthened tobacco control policies.

What is already known about this topic?:The prevalence of metabolic multimorbidity is rising in China and has been linked to lifestyle factors; however, evidence characterizing the dynamic transitions across disease states and the stage-specific influence of lifestyle behaviors remains scarce. What is added by this report?:At baseline, the prevalence of metabolic multimorbidity was 24.14%, with males, older adults, and individuals with adverse lifestyle behaviors bearing a disproportionately elevated risk. Hepatic steatosis was associated with a markedly increased risk of progression to multimorbidity, while smoking, high-salt diets, and irregular physical activity each showed differential associations with specific disease transition pathways. What are the implications for public health practice?:Lifestyle behaviors exert meaningful influence at every stage of metabolic disease progression, identifying them as priority targets for stage-specific prevention and control strategies in China.

AIMS:To examine the associations between early-pregnancy maternal overweight/obesity metabolic phenotypes and foetal anthropometric measurements and assess mediation by gestational diabetes mellitus (GDM) and pregnancy-induced hypertension (PIH). METHODS:This study included 7633 pregnant women from a Chinese prospective cohort. Participants in early pregnancy were classified as metabolically healthy normal weight (MHNW), metabolically unhealthy normal weight (MUNW), metabolically healthy overweight/obesity (MHO) and metabolically unhealthy overweight/obesity (MUO). Generalised linear mixed models evaluated associations with repeated ultrasound measures for participants with at least one scan between 22 and 36 weeks. Mediation analyses evaluated GDM and PIH effects. RESULTS:Compared to MHNW, both MHO and MUO were associated with higher estimated foetal weight (EFW) and abdominal circumference (AC) Z-scores mainly during 28-36 weeks, as well as an elevated risk of foetal EFW and AC overgrowth. Furthermore, the MUO increased the risk of head circumference (HC) undergrowth (OR = 1.22, 95% CI: 1.00-1.48). Conversely, the MUNW was associated with lower foetal HC Z-scores (β = -0.05, 95% CI: -0.10 to 0.00). In addition, GDM mediated the association between MUNW and reduced foetal HC Z-scores at 28-32 and 33-36 weeks, with mediation proportions of 11.2% and 16.1%, respectively. CONCLUSIONS:Early-pregnancy maternal overweight/obesity metabolic phenotypes are associated with foetal growth. MHO and MUO increase EFW and AC overgrowth risks, while MUO also raises HC undergrowth risk. Conversely, MUNW is associated with a reduced HC Z-score, a pathway mediated by GDM. These findings highlight that the joint assessment of body mass index and metabolic health is vital for targeted maternal care.

Genome-wide association studies (GWASs) have identified at least 17 genetic variants associated with gastric cancer risk. However, the underlying genetic regulatory mechanisms remain poorly understood. We systematically performed genome-wide analyses of expression quantitative trait loci (eQTLs), splicing QTLs (sQTLs), and alternative polyadenylation QTLs (apaQTLs) using gastric biopsy tissues from 262 Chinese individuals, and we integrated these data with GWAS results from 10,254 gastric cancer cases and 10,914 controls. We identified 4,636 eGenes, 1,422 sGenes, and 511 apaGenes, with 55.3% of sGenes and 49.5% of apaGenes being distinct from eGenes. Notably, over half of the variants associated with shared genes were unique to a specific QTL type, with these variants characterized by a wide physical separation (median distance = 25.01 kb) and weak linkage disequilibrium (median r2 = 0.42). Gastric cancer risk variants showed significant enrichment across all three QTL categories. Integrative transcriptome-wide association studies (TWASs) across expression, splicing, and alternative polyadenylation identified 34 genes at false discovery rate (FDR) <0.05; PSCA was implicated across all three regulatory layers. The candidates DIP2B and RMC1 were experimentally validated to promote gastric tumorigenesis both in vitro and in vivo. Functional analyses revealed that the rs11264361-T allele reduced FDPS exon 9 skipping, leading to increased expression of the full-length transcript, while the apaVariant rs7445 regulated UBE2L3 polyadenylation via ATXN2 binding, contributing to gastric cancer risk. Collectively, these results provide a comprehensive QTL resource in gastric tissues and demonstrate distinct regulatory mechanisms of eQTLs, sQTLs, and apaQTLs in gastric cancer development.

RATIONALE:During tumorigenesis, approximately 20% of EGFR-mutant lung adenocarcinomas (LUADs) progress rapidly to aggressive subtypes. Multi-omics analyses of stage I LUAD cohorts have revealed that centrally located lesions exhibit enhanced tumorigenic potential compared with peripheral counterparts, whereas the underlying mechanisms remain elusive. OBJECTIVES:To define the spatial-clinical determinants of early aggressive progression in EGFR-mutant LUAD and develop a lineage-based mechanistic framework connecting regional microenvironmental constraints with epithelial cell-state remodeling and invasive acquisition. METHODS:We conducted an integrated multi-omics analysis combining clinical cohort data (n = 277), single-cell and spatial transcriptomics, and functional studies in genetically engineered mouse models to identify spatial-clinical patterns. Mechanistic studies were carried out using mouse models, 3D organoids, and controlled oxygen interventions to investigate the effects of hypoxia on cellular transformation. MEASUREMENTS AND MAIN RESULTS:Our analysis revealed that centrally located lesions display enhanced tumorigenic potential compared to peripheral counterparts, driven by hypoxic niche. Hypoxic preconditioning (10% O2) induced ribosome collisions in EGFR-driven mouse models and organoids, activating the ZAKα-MAPK-c-Fos axis to disrupt alveolar lineage imbalance, characterizing as suppressing alveolar epithelial factor NKX2-1 while elevating stem-like progenitor FOXD1. Therapeutic hyperoxia (60% O2) restored lineage balance and attenuated tumorigenesis. CONCLUSIONS:A preexisting hypoxic niche is a key spatial determinant of early malignant progression in EGFR-mutant LUAD, engaging ribosome-collision signaling through the ZAKα-MAPK-c-Fos axis and promoting alveolar lineage imbalance. Restoring oxygenation, either through controlled hyperoxia or pharmacologic inhibition of this pathway, may help curb tumorigenesis and rapid progression of centrally located lesions.

Preterm infants are susceptible to metabolic disruptions due to physiologically immature development, and early metabolic dysregulation may contribute to neurodevelopmental impairments that persist throughout infancy and beyond. This study aims to investigate the associations between preterm birth, neonatal metabolism, and later neurodevelopment, and to explore the potential mediating role of neonatal metabolism. In this prospective birth cohort of 9023 in China, linear regression analyses were employed in discovery and validation sets to identify metabolites associated with preterm birth. Metabolites were then categorized as extremely high (> 90th percentile) or low (< 10th percentile), and their associations with preterm birth were assessed using meta-analysis and logistic regression. Among 2086 infants with neurodevelopmental assessments at 1 year old, we applied restricted cubic splines and linear regression to evaluate associations between extreme metabolite levels and neurodevelopment. Mediation analysis was then performed to assess the potential mediating effects of neonatal metabolism. Preterm birth was associated with extremely low levels of four metabolites and extremely high levels of seven metabolites (e.g., 17-hydroxyprogesterone, alanine, and multiple carnitines/acylcarnitines), indicating perturbed neonatal metabolic profiles. Moreover, extremely high levels of free carnitine (C0) were associated with poorer cognition (β = −0.47; 95

PurposeInfants born after assisted reproductive technology (ART) have more adverse health outcomes than spontaneously conceived infants. Evidence on the impact of ART on the offspring's visual system, particularly refractive errors (REs), remains limited and inconsistent.MethodsThe present study included 6847 infants from the Jiangsu Birth Cohort (JBC), which enrolled couples who conceived either through ART or spontaneously between 2015 and 2022 in Jiangsu Province, China. ART procedures and parental characteristics were collected prospectively, and REs were assessed at one year of age using the Spot Vision Screener. Poisson regression models were used to examine the association between ART and screening-detected REs.ResultsScreening-detected REs were identified in 369 infants born after ART (11.4%) and in 405 infants spontaneously conceived (11.2%). No overall association was observed between ART and the risk of REs (RR, 0.91; 95% CI 0.77-1.07). Infants born to parents with different infertility diagnoses or following specific ART procedures had similar risks of REs compared with spontaneously conceived infants. In the analyses restricted to ART-conceived infants, infants conceived using the gonadotropin-releasing hormone antagonist (GnRH-antagonist) protocol had a lower risk of REs (RR, 0.56; 95% CI 0.44-0.71) compared with those using the gonadotropin-releasing hormone agonist (GnRH-agonist) protocol.ConclusionWe observed no increased risk of screening-detected REs at one year of age in ART-conceived infants compared with spontaneously conceived infants, independent of parental infertility diagnoses or specific ART procedures. Among the ART-conceived infants, the use of the GnRH-antagonist protocol may be associated with a lower risk of screening-detected REs; however, this finding should be interpreted cautiously because protocol selection was clinically guided and residual confounding cannot be excluded.

ObjectiveThis study analyzed SARS-CoV-2 surveillance results in the urban wastewater of Zhengzhou, central China, to explore its association with community COVID-19 incidence and evaluate its early warning performance across different SARS-CoV-2 variant epidemic phases.MethodsWeekly monitoring was conducted at five representative wastewater treatment plants from May 2024 to July 2025, yielding 325 samples. Viral loads of the ORF1ab and N genes were quantified using RT-qPCR, with statistical analyses performed included Spearman correlation, ANOVA, and cross-correlation function.ResultsThe two target genes showed high consistency and significant positive correlation with 14-day cumulative COVID-19 incidence. However, significant variant and marked heterogeneity was observed across variant periods and gene targets, with the N gene outperforming ORF1ab in stability. Wastewater viral signals preceded clinical cases by one week.ConclusionThese findings supported the application value of wastewater-based epidemiology in post-pandemic COVID-19 surveillance, indicate its potential early warning capacity, and reveal notable heterogeneity in surveillance performance across viral variants and target genes.

BACKGROUNDS:The impact of maternal gestational diabetes mellitus (GDM) and its glycemic subtypes on the early visual development of infants remains unclear. METHODS:Based on the Jiangsu Birth Cohort, we included 2139 infants born to 2041 mothers recruited from 2014 to 2018. Maternal GDM was categorized into three categories based on oral glucose tolerance test results: impaired fasting glucose, impaired glucose tolerance, and comorbid impaired fasting glucose and impaired glucose tolerance. Infants' grating visual acuity was measured at 1 year of age using Teller Acuity Cards II. We used generalized estimating equation models to examine the association between maternal GDM and infant visual acuity, accounting for the dependence of twin observations. We then conducted a metabolomics analysis to explore the metabolic profiles associated with GDM and infant visual acuity. RESULTS:Prenatal GDM exposure was associated with a 70% increase in risk of abnormal visual acuity in infants (relative risks [RR] = 1.7; 95% confidence interval [CI] = 1.2, 2.3). The increased risks were more pronounced among infants in the comorbid impaired fasting glucose and impaired glucose tolerance group (RR = 3.2; 95% CI = 1.4, 7.7), followed by the impaired glucose tolerance group (RR = 1.8; 95% CI = 1.1, 3.0) and the impaired fasting glucose group (RR = 1.6; 95% CI = 0.7, 3.6), compared with unexposed infants. The metabolomics analysis suggested glycine, serine, and threonine metabolism as an enriched pathway and identified creatine as a metabolite of interest. CONCLUSIONS:Maternal GDM is associated with an increased risk of abnormal grating visual acuity in infants at 1 year of age. Glycine, serine, and threonine metabolism pathways may contribute to this association.

Gestational diabetes mellitus (GDM) is a heterogeneous condition diagnosed solely through glucose. It is characterized by profound perturbations in the metabolome, with specific metabolic profiles linked to adverse birth outcomes. Metabolomics can reveal population heterogeneity in health and disease. Here, we employed non-targeted metabolomics to systematically profile the circulating metabolome in 2050 pregnant women during mid-pregnancy, identifying 30 metabolites that define the GDM metabolic signature (mGDM). Participants were stratified into four groups by distinct glycemic and metabolic profiles, namely normoglycemic-non-mGDM, hyperglycemic-non-mGDM, normoglycemic-mGDM, and hyperglycemic-mGDM, and associations with subsequent adverse birth outcomes were assessed. Compared with normoglycemic-non-mGDM, normoglycemic-mGDM demonstrated nearly a 2-fold increased risk of preterm birth (OR 1.93, 95% CI 1.02–3.65) and large for gestational age (OR 2.11, 95% CI 1.53–2.92). Conversely, hyperglycemic-non-mGDM did not show elevated risks in adverse birth outcomes versus normoglycemic individuals. Hyperglycemic-mGDM individuals were associated with higher risks of preterm birth (OR 2.37, 95% CI 1.04–5.39), large for gestational age (OR 2.28, 95% CI 1.50–3.47), congenital malformations (OR 1.87, 95% CI 1.03–3.39), and neonatal intensive care unit admissions (OR 1.69, 95% CI 1.09–2.61). We observed a stepwise increase in adverse outcome risk across the four-level metabolic-glycemic categories (P for trend<0.001 for large for gestational age; 0.013 for preterm birth; 0.018 for NICU admission). Taken together, our study outlines the metabolic profile of GDM and reveals clinically relevant heterogeneity in adverse pregnancy outcomes by metabolic signature. Integrating blood glucose and metabolomics may improve risk stratification and advance precision maternal care.

The TBX1 deletion is frequently observed in patients with tetralogy of Fallot (TOF), the most prevalent cyanotic congenital heart defect (CHD); however, the role of noncoding variants in its regulatory region remains unclear. We performed whole-genome sequencing (WGS) on a cohort of 428 patients diagnosed with TOF, which identified the presence of 22q11.2 deletion in 31 cases, including the TBX1 genomic region, as well as 7 functional noncoding variants in its enhancer (EnhTBX1). These findings accounted for a total of 9.81% (42/428) of TOF cases. We then knocked out EnhTBX1 (EnhTBX1-/-) in ES cells and utilized blood vessel organoids (BVOs) as a model to elucidate the mechanisms underlying outflow tract (OFT) vessel dysfunction. The EnhTBX1-/- BVOs exhibited significantly reduced TBX1 mRNA expression, leading to impaired angiogenesis, vessel regression with decreased pericyte coverage, and alterations in tight junction morphology. Mechanistically, we have identified TBX1 as a specific transcription factor for endothelial progenitor cells that influences their differentiation. The main targets of TBX1 are genes involved in angiogenesis-related pathways, such as DLL4 and TGFBR2, which have been reported to play crucial roles during OFT formation. Finally, rescue experiments with TBX1 overexpression not only reinstated the expression of aforementioned target genes but also recovered tight junction formation and enhanced pericyte coverage rate of EnhTBX1-/- BVOs. In conclusion, this study demonstrates the involvement of noncoding variants in the TBX1 enhancer in TOF pathogenesis, highlighting the feasibility of using organoids to decipher the underlying mechanisms of unconserved noncoding regions in developmental malformations.

Lung cancer in smokers (LCIS) and lung cancer in never-smokers (LCINS) are different entities with distinct molecular features. However, their cellular heterogeneity still requires further investigation. Through an integrated analysis of single-cell RNA sequencing and bulk sequencing data, we identified cell subpopulations associated with smoking and non-smoking patients. Subsequent transcriptomic analyses were performed to elucidate differences in cellular functions and the tumor microenvironment. We observed that smoking-associated cancer cells exhibited a higher degree of aggressiveness, which may correlate with an adverse prognosis in smoking patients. Additionally, immunosuppressive CXCL10 + macrophages may contribute to tumorigenesis in smokers, and the immunoregulatory LGALS9-HAVCR2 axis could be a potential immunotherapeutic target. In non-smokers, the inflammatory microenvironment may be involved in tumor development. Moreover, the reduced anti-tumor cytotoxicity may be associated with their suboptimal immunotherapeutic response. Our study uncovered differences in oncogenic and immune escape mechanisms between LCIS and LCINS patients and suggests potential immunotherapeutic strategies.

De novo mutations (DNMs) are an important source of congenital diseases. With delayed parenthood and assisted reproductive technology (ART) use increasing, it is essential to elucidate how these reproductive factors influence DNMs and whether resulting mutations influence offspring health. Here we performed whole-genome sequencing of 24,030 individuals from 7,851 parent-offspring families, identifying 390,924 de novo single-nucleotide variants (dnSNVs). Paternal and maternal aging exhibited distinct mutational patterns, with maternal DNM accumulation accelerating at advanced ages. Increased paternal dnSNVs partially accounted for the association between advanced parental age and shorter gestational duration. Moreover, ART showed age-independent, procedure-specific effects: intracytoplasmic sperm injection (ICSI) and ovarian stimulation were associated with increased paternal and maternal dnSNVs, respectively, and ICSI-associated paternal dnSNVs also partially accounted for the association between ICSI and shorter gestational duration. In vitro embryo manipulation was associated with increased early post-zygotic mosaic mutations, particularly C > A substitutions linked to delayed neurocognitive development at 1 year. Collectively, these findings advance understanding of the determinants and consequences of de novo mutagenesis.

Colorectal cancer (CRC) remains a major global health concern, underscoring the need for reliable biomarkers for diagnosis and prognosis. In this study, untargeted plasma proteomics was conducted across two cohorts comprising 321 participants in the discovery set and 353 in the validation set. Machine learning methods were employed to select biomarkers and develop diagnostic and prognostic models, with performance evaluated by the area under the receiver operating characteristic curve (AUC). An eight-protein panel for CRC and a six-protein panel for advanced adenoma discriminated from healthy controls with AUCs of 0.932 and 0.816 in the validation set, respectively. Additionally, a combined prognostic model incorporating eight proteins and clinical factors predicted 5-year disease-free survival in CRC with an AUC of 0.744 in the validation cohort. Overall, the identified protein biomarkers have the potential to foster the development of effective blood-based tests for early detection and prognostic prediction of CRC.

Recurrent implantation failure (RIF), characterized by repeated failure to achieve clinical pregnancy after embryo transfer, is often associated with abnormal endometrial conditions. However, unexplained RIF is unique in that its underlying cause remains largely unknown, posing a challenge for both clinicians and patients. We performed single-cell RNA sequencing and reported that the endometria of patients with unexplained RIF exhibited increased the proliferation and activation of cytotoxic CD8+ T-cells, which hinder embryo implantation. Additionally, the glandular epithelium, which has the highest metabolic activity, showed abnormal glycolysis and reduced lactate production in RIF. Specifically, the endometrial expression of genes associated with glucose uptake (SLC2A1), glucose metabolism (ALDOA), lactate production (LDHA), and lactate output (SLC16A3) were lower in the patients with unexplained RIF than in the controls. Through uterine horn injection experiments in mice, we demonstrated that inhibiting lactate production in the endometrium prevents embryo implantation and that this effect could be reversed by lactate supplementation. Moreover, lactate inhibitors did not affect implantation in mice with CD8+ T-cells depletion. In vitro experiments also confirmed that lactate inhibition affects the proliferation and activation of CD8+ T-cells. We propose that the endometria of patients with unexplained RIF fail to establish proper immune balance toward the embryo, likely due to abnormal glycolysis and reduced lactate production in the glandular epithelium.

INTRODUCTION:Smoking is a strong modifiable prognostic factor for lung cancer survival. We compared eight smoking metrics to determine which metric best models the relationship between smoking exposure with overall survival (OS) and lung cancer-specific survival (LCSS). These metrics included cigarettes-per-day, smoking duration, pack-years, square-root pack-years, logcig-years, the comprehensive smoking index, age-of-initiation, and years-since-quit. MATERIALS/METHODS:This retrospective, pooled analysis included 25 International Lung Cancer Consortium studies between June 1, 1983-December 31, 2019. The performance of smoking metrics for modelling OS was compared based on 1) strength and significance in adjusted Cox-proportional hazard models and 2) linearity based on the goodness-of-fit assuming the log-hazard varies linearly with each smoking metric (i.e. the hazard ratio is constant across different values of the smoking metric) compared to models using non-linear splines. This process was repeated across clinicodemographic subgroups and for LCSS. RESULTS:In total, 28,702 lung cancer patients were included (median age 64 [IQR: 57-71]; 53% male). Logcig-years (log(cigarettes/day+1)·years-smoked) had the highest adjusted hazard ratio per standard deviation (aHR 1.11; 95% CI: 1.09-1.13) and best goodness-of-fit when modelled linearly. Square-root pack-years had a similar effect size (aHR 1.11; 95% CI: 1.09-1.13) and had a strong linear relationship on visual assessment of spline curves. In subgroup analyses, logcig-years had a large effect size and maintained a linear relationship regardless of age, sex, stage, and histology. For lung cancer-specific survival (LCSS), logcig-years again had the highest aHR (1.09; 95% CI: 1.05-1.12) and the best linear goodness-of-fit, while square-root pack-years demonstrated the most linear relationship on visual assessment. DISCUSSION:Logcig-years best modelled the relationship between smoking exposure and OS as well as LCSS, and had consistent associations across clinicodemographic subgroups. Logcig-years should be considered in clinical and research applications for quantifying smoking exposure in lung cancer.

Liver function-related quantitative biomarkers (LFQBs) are essential for assessing hepatic health, yet prior genome-wide association studies (GWAS) have largely studied them in isolation. We conducted cross-ancestry GWAS meta-analyses on seven LFQBs to further elucidate liver function’s genetic architecture, identify pleiotropic variants, and prioritize genes to pinpoint targets with therapeutic potential. We performed GWAS meta-analyses on seven LFQBs in 456,000 individuals across East Asian, European, South Asian, and African ancestries, followed by a series of downstream analyses, including fine-mapping, phenome-wide association study (PheWAS), gene-linking, and Mendelian randomization (MR). We identified 5,507 lead signals (P < 5 × 10−9), including 210 novel ones. Fine-mapping revealed 2,012 putative causal variants, of which 38 concurrently exhibited causal signals across multiple LFQBs and showed widespread associations with liver- and metabolism-related traits in PheWAS. Additionally, polygenic risk score (PRS)-based PheWAS uncovered pan-systemic manifestations of hepatic homeostasis disruption across diverse phenotypic domains. We proposed a novel multidimensional gene-linking framework (mdS2G) to bridge the identified causal loci to 1,166 putative genes. Benchmarking analysis revealed that mdS2G exhibited superior hepatocyte enrichment compared to individual constituent strategies. Furthermore, MR analysis highlighted PEPD protein as exhibiting therapeutic potential for metabolic dysfunction-associated steatotic liver disease (MASLD) and cirrhosis (P = 2.24 × 10−5, 4.36 × 10−3, respectively). This study maps the genetic landscape of liver function with expanded ancestral diversity, uncovering key variants and genes tied to hepatic homeostasis disruption and laying the groundwork for targeted liver disease therapies. Additionally, our open-access gene-linking framework provides a resource for the rapid prioritization of putative genes to guide future experimental interrogation.