Background/Aims: Liver transplantation (LT) following total hepatectomy is a life-saving treatment for hepatocellular carcinoma (HCC). The HCC recurrence after LT hinders the effectiveness of the procedure. The objective of this study is to develop a pre-operative risk stratification model based on a liquid biopsy. Methods: We conducted a comprehensive multi-omics study of 260 HCC patients from three centers, including clinical data, low-coverage whole-genome sequencing of cell-free DNA (cfDNA) from plasma, as well as whole-exome, single-nucleus RNA, and spatial transcriptomics from matched tumor and non-tumor tissues. Results: We identified cfDNA-derived copy number alteration (CNA) signatures associated with post-transplant recurrence. By integrating cfDNA-derived CNA profiles with single-cell transcriptomic data, we traced recurrence-associated cfDNA to a distinct subpopulation of malignant cells within the primary tumor. These cells were embedded in a pro-metastatic microenvironment of specialized endothelial subtypes and cancer-associated fibroblasts. Notably, most recurrence-associated lesions were detectable in cfDNA prior to liver transplantation (LT). Building on these insights, we developed the ZJU Criteria based on CNA fragments and tumor markers, a pre-LT risk prediction tool that integrates conventional clinical factors with cfDNA-derived CNA signatures, and validated it using internal and independent external cohorts. Conclusion Our findings suggest that post-transplant recurrence commonly originates from advanced subclones that emerge late during tumor evolution. The ZJU Criteria provides an accurate, non-invasive strategy that significantly improves pre-LT risk stratification and clinical decision-making for patients with HCC.

ObjectiveMigraine is a leading neurological disorder and the fourth most common cause of years lived with disability worldwide, affecting nearly 116 million individuals. Although pharmacological treatments are available, their efficacy is often limited by side effects and variable response rates. Repetitive transcranial magnetic stimulation (rTMS) over the dorsolateral prefrontal cortex (DLPFC) offers a safe, non-invasive alternative for migraine management. However, the neurophysiological mechanisms, particularly how rTMS modulates local cortical excitability and distributed pain-related circuits, remain poorly understood. Elucidating these mechanisms is essential for optimizing treatment protocols and improving clinical outcomes. MethodsThis study employed concurrent transcranial magnetic stimulation and electroencephalography (TMS-EEG) to investigate neuroplastic and neurocircuitry mechanisms of DLPFC-rTMS in migraine. Study 1 compared 30 migraineurs and 28 healthy controls to identify abnormalities in TMS-evoked potentials (TEPs) and significant current density (SCD) within sensory-discriminative regions including the primary somatosensory cortex (S1) and posterior insula (pINS), cognitive-affective regions including the anterior insula (aINS) and midcingulate cortex (MCC), and a descending modulatory region, the periaqueductal gray (PAG). Study 2 used a single-blind, crossover, sham-controlled design in 34 healthy participants. Each participant received both active (10 Hz, 80% RMT, 1 500 pulses) and sham DLPFC-rTMS in counterbalanced order. TMS-EEG and cold pain tolerance were assessed before and after each session. ResultsIn Study 1, migraineurs showed a significantly less negative N120 amplitude compared to healthy controls (P=0.027, Cohen’s d=0.60), indicating local intracortical disinhibition. No group differences were observed for N40, P60, or P180 components. At the source level, migraineurs exhibited significantly higher SCD in the S1, pINS, aINS, and MCC (allQ<0.05), but not in the ventroposterior thalamus (vpTHAL), mediodorsal thalamus (mdTHAL), or PAG. In Study 2, active rTMS significantly reduced SCD from pre- to post-stimulation in the S1, aINS, and MCC (all Q<0.05). Sham stimulation also reduced SCD in the S1 (Q<0.05) but not in the aINS or MCC. Although no significant group-level analgesic effect was observed between active and sham conditions (P=0.107), correlation analyses revealed that greater SCD reductions in the S1 and MCC were significantly associated with higher post-rTMS pain tolerance (R=-0.487 and -0.495, both Q<0.01) and larger improvements in pain tolerance(R=-0.487 and -0.451, both Q<0.05). No such correlations were found following sham stimulation, suggesting that the behavioural relevance of neural changes is specific to active rTMS. ConclusionThis study provides novel evidence that migraineurs exhibit both local neuroplastic abnormalities (reduced N120 amplitude) and hyperactivity in key pain-processing regions (S1, pINS, aINS, MCC). A single session of DLPFC-rTMS reduced hyperactivity in the aINS, MCC, and S1. Notably, greater reductions in the S1 and MCC were associated with improved pain tolerance. These findings identify distinct cortical circuitries, particularly within the cognitive-affective pain network, that may serve as potential biomarkers for optimizing rTMS treatment in migraine and other chronic pain conditions. Future studies should validate these results in patient populations experiencing spontaneous migraine attacks and explore multi-session or accelerated rTMS protocols.

The general models for intermediates quality analysis in the production process of Yaobitong capsule were established by near infrared spectroscopy (NIRS) combined with chemometrics, realizing the rapid determination of notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, ginsenoside Rd and moisture. The spray-dried fine powder and total mixed granule were selected as research objects. The contents of five saponins were determined by high performance liquid chromatography and the moisture content was determined by drying method. The measured contents were used as reference values. Meanwhile, NIR spectra were collected. After removing abnormal samples by Monte Carlo cross validation (MCCV), Monte Carlo uninformative variables elimination (MC-UVE) and competitive adaptive reweighted sampling (CARS) were used to select feature variables respectively. Based on the feature variables, quantitative models were established by partial least squares regression (PLSR), extreme learning machine (ELM) and ant lion optimization least squares support vector machine (ALO-LSSVM). The results showed that CARS-ALO-LSSVM model had the optimum effect. The correlation coefficients of the six index components were greater than 0.93, and the relative standard errors were controlled within 6%. ALO-LSSVM was more suitable for a large number of samples with rich information, and the prediction effect and stability of the model were significantly improved. The general models with good predicting effect can be used for the rapid quality determination of Yaobitong capsule intermediates.

Background: The genetic basis for hyperglycaemia in pregnancy remain unclear. This study aimed to uncover the genetic determinants of gestational diabetes mellitus (GDM) and investigate their applications. Methods: We performed a meta-analysis of genome-wide association studies (GWAS) for GDM in Chinese women (464 cases and 1,217 controls), followed by de novo replications in an independent Chinese cohort (564 cases and 572 controls) and in silico replication in European (12,332 cases and 131,109 controls) and multi-ethnic populations (5,485 cases and 347,856 controls). A polygenic risk score (PRS) was derived based on the identified variants. Results: Using the genome-wide scan and candidate gene approaches, we identified four susceptibility loci for GDM. These included three previously reported loci for GDM and type 2 diabetes mellitus (T2DM) at MTNR1B (rs7945617, odds ratio [OR], 1.64; 95% confidence interval [CI],1.38 to 1.96]), CDKAL1 (rs7754840, OR, 1.33; 95% CI, 1.13 to 1.58), and INS-IGF2-KCNQ1 (rs2237897, OR, 1.48; 95% CI, 1.23 to 1.79), as well as a novel genome-wide significant locus near TBR1-SLC4A10 (rs117781972, OR, 2.05; 95% CI, 1.61 to 2.62; Pmeta=7.6×10-9), which has not been previously reported in GWAS for T2DM or glycaemic traits. Moreover, we found that women with a high PRS (top quintile) had over threefold (95% CI, 2.30 to 4.09; Pmeta=3.1×10-14) and 71% (95% CI, 1.08 to 2.71; P=0.0220) higher risk for GDM and abnormal glucose tolerance post-pregnancy, respectively, compared to other individuals. Conclusion Our results indicate that the genetic architecture of glucose metabolism exhibits both similarities and differences between the pregnant and non-pregnant states. Integrating genetic information can facilitate identification of pregnant women at a higher risk of developing GDM or later diabetes.

Background: The genetic basis for hyperglycaemia in pregnancy remain unclear. This study aimed to uncover the genetic determinants of gestational diabetes mellitus (GDM) and investigate their applications. Methods: We performed a meta-analysis of genome-wide association studies (GWAS) for GDM in Chinese women (464 cases and 1,217 controls), followed by de novo replications in an independent Chinese cohort (564 cases and 572 controls) and in silico replication in European (12,332 cases and 131,109 controls) and multi-ethnic populations (5,485 cases and 347,856 controls). A polygenic risk score (PRS) was derived based on the identified variants. Results: Using the genome-wide scan and candidate gene approaches, we identified four susceptibility loci for GDM. These included three previously reported loci for GDM and type 2 diabetes mellitus (T2DM) at MTNR1B (rs7945617, odds ratio [OR], 1.64; 95% confidence interval [CI],1.38 to 1.96]), CDKAL1 (rs7754840, OR, 1.33; 95% CI, 1.13 to 1.58), and INS-IGF2-KCNQ1 (rs2237897, OR, 1.48; 95% CI, 1.23 to 1.79), as well as a novel genome-wide significant locus near TBR1-SLC4A10 (rs117781972, OR, 2.05; 95% CI, 1.61 to 2.62; Pmeta=7.6×10-9), which has not been previously reported in GWAS for T2DM or glycaemic traits. Moreover, we found that women with a high PRS (top quintile) had over threefold (95% CI, 2.30 to 4.09; Pmeta=3.1×10-14) and 71% (95% CI, 1.08 to 2.71; P=0.0220) higher risk for GDM and abnormal glucose tolerance post-pregnancy, respectively, compared to other individuals. Conclusion Our results indicate that the genetic architecture of glucose metabolism exhibits both similarities and differences between the pregnant and non-pregnant states. Integrating genetic information can facilitate identification of pregnant women at a higher risk of developing GDM or later diabetes.

ObjectiveTo assess the current status of medical waste management in Jinshan District of Shanghai, China, to identify existing issues, and to provide a scientific basis for formulating targeted strategies. MethodsData were collected from the routine supervision and inspection records of the Jinshan District Health Commission Supervision Institute from 2017 to 2021, covering all aspects of medical waste management, including collection, classification, transportation, storage, and administrative penalties. ResultsThe compliance rates for the establishment of institutional frameworks, staffing, internal handover, and registration in medical and healthcare institutions all exceeded 95.00%. However, only 2.31% of the medical and healthcare institutions met the 48-hour storage limit requirement for medical waste. Private institutions had significantly lower compliance rates (P<0.05) in aspects such as proper classification and collection, maintaining records for three years, adhering to the 48-hour storage limit, refraining from commercial transactions, timely disinfection and cleaning, and implementing emergency measures for waste loss. Compliance rates also varied among different types of institutions regarding the establishment of temporary storage facilities and the implementation of the transfer manifest system, with community healthcare institutions exhibiting relatively lower compliance rates (P<0.05). Over the past five years, private medical and healthcare institutions accounted for 63.33% of administrative penalty cases. ConclusionWhile medical waste management in Jinshan District, Shanghai, has gradually become more standardized, challenges remain. To address the issue of medical waste being stored for over 48 hours, medical waste transfer stations should be established to improve transfer efficiency and ensure complete waste collection. Additionally, for private and community healthcare institutions, weak links in management should be addressed by establishing medical waste quality control teams, enhancing supervision through digital tools, and optimizing management processes to comprehensively elevate medical waste management.

The incidence of male infertility has been progressively increasing, its pathogenesis still incompletely elucidated and associated with multifactorial etiologies. Among these, sexually transmitted infections (STIs) constitute a significant contributing factor to male infertility. STIs, caused by various pathogens including human papillomavirus (HPV), may impair male fertility through diverse pathophysiological mechanisms. Nevertheless, current investigations into the impact of HPV infection on male reproductive capacity remain limited. This review critically examines the effects of HPV infection on the anatomical and functional integrity of the male reproductive system, semen parameters, and sperm quality, while exploring the correlation between HPV-related clinical management and male fertility preservation. Through comprehensive elucidation of HPV's pathogenic mechanisms within the male reproductive system and identification of its potential etiological contributions to male infertility, this review aims to provide theoretical foundations for optimizing fertility guidance protocols and developing novel antiviral therapeutic interventions, thereby advancing clinical strategies for reproductive health management and inform future research directions in antiviral treatment development.

Objective To investigate and compare the epidemiological characteristics of common respiratory viruses among influenza-like illness(ILI)and severe acute respiratory infection(SARI)cases in Huzhou,Zhejiang Province before and after the COVID-19 pandemic,so as to provide a basis for formulating and adjusting the prevention and control strategies for viral respiratory infectious diseases.Methods ILI and SARI cases at two influenza surveillance sentinel hospitals in Huzhou and had throat swab samples collected during Nov 2017 to Feb 2020(pre-COVID-19 pandemic period)and Dec 2022 to Apr 2024(post-COVID-19 mitigation phase)were selected as the participants.Seven common viral respiratory pathogens were tested,including influenza A virus(H1N1 and H3N2 subtypes),influenza B virus(Victoria lineage,FluB),respiratory syncytial virus(RSV),rhinovirus(HRV),adenovirus(ADV),and severe acute respiratory syndrome coronavirus-2(SARS-CoV-2).The positive rates of respiratory pathogens before and after the COVID-19 pandemic were compared across different age groups and different time.Results A total of 7 948 ILI samples and 2 294 SARI samples were included.The overall positive rate of ILI samples increased from 33.6%to 47.1%,primarily due to the increase in influenza and COVID-19 infections;the overall positive rate of SARI samples decreased from 31.4%to 24.8%,mainly due to the reduction in HRV and ADV infections.During the post-COVID-19 mitigation phase,SARS-CoV-2(22.1%),H3N2(12.7%),and FluB(6.0%)were the primary pathogens in ILI samples,while RSV(7.1%),H3N2(5.3%),and HRV(4.5%)dominated in SARI samples.During the post-COVID-19 mitigation phase,the influenza virus circulation period was shortened.Before the COVID-19 pandemic,RSV was mainly detected in autumn and winter,while during the post-COVID-19 mitigation phase,out-of-season RSV epidemics were observed in spring and summer.Co-infection rate in ILI cases increased significantly in the post-COVID-19 mitigation phase,predominantly consisting of co-infections of COVID-19 and influenza A virus,while co-infection rate in SARI cases showed a decline.Conclusion We found important epidemiological changes in respiratory viruses in Huzhou during the post-COVID-19 mitigation phase compared to pre-COVID-19 period,including increased positive rates of influenza and COVID-19,and disruptions to the seasonal patterns of influenza and RSV.The prevention and control strategies should be adjusted in a timely manner based on the monitoring data.

Objective:To investigate the ocular pathogen spectrum and drug sensitivity characteristics of patients to be diagnosed with infectious keratitis within 10 years in two tertiary hospitals in Nantong City, Jiangsu Province, eastern China.Methods:A cross-sectional study was conducted.Microbial culture specimens from consecutive 1 404 patients with suspected infectious keratitis admitted to the Department of Ophthalmology at the First People's Hospital of Nantong City and Nantong University Affiliated Hospital from January 2014 to October 2023 were collected.The patients' general data, etiology and drug sensitivity test results were analyzed.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Nantong First People's Hospital (No.2021KT273) and Nantong University Affiliated Hospital (No.2019-K068).Results:Among 1 404 patients with suspected infectious keratitis, the positive rate of microbial culture was 37.04%(520/1 404).A total of 537 strains were isolated and cultured, with fungi accounting for 69.09%(371/537) and bacteria accounting for 30.91%(166/537).The common bacterial genera in fungal keratitis were Fusarium (47.17%, 175/371), Alternaria (15.90%, 59/371), Aspergillus (14.56%, 54/371), Certospora (10.78%, 40/371) and Penicillium (3.50%, 13/371).The annual composition ratio of Fusarium showed a downward trend, while Certospora showed an upward trend.The common bacteria in bacterial keratitis were Staphylococcus epidermidis (24.10%, 40/166), Streptococcus pneumoniae (17.47%, 29/166), Pseudomonas aeruginosa (13.25%, 22/166), Staphylococcus aureus (6.63%, 11/166) and Corynebacterium macginleyi (4.22%, 7/166).The annual composition ratio of Pseudomonas aeruginosa showed an increasing trend.The resistance rates of gram-positive bacteria to levofloxacin and vancomycin were 36.26% and 0% respectively, and the resistance rates of gram-negative bacteria to aminoglycosides and ceftazidime were <10%.A total of 61 bacterial strains (40.94%) were multi-drug resistant. Conclusions:Fusarium is common in fungal keratitis, and Corynebacterium macginleyi in bacterial keratitis may be a microbial feature in Nantong City.Levofloxacin may no longer be suitable as a first-line antibiotic for topical ocular use.

Objective Exploring the mechanism of mitophagy-mediated efferocytosis in myocardial fibrosis progression based on transcriptomics and bioinformatics,with potential chinese herbal medicine prediction and experimental validation.Methods A rat model of myocardial fibrosis was constructed by injecting angiotensin Ⅱ subcutaneously.RNA extraction and high-throughput sequencing were performed on myocardial tissue of rats,and differentially expressed genes(DEGs)were detected.Efferocytosis-related genes(ERGs)were obtained from the GeneCards database,and the intersection of DEGs and ERGs was mapped using the"venneuler"package in R 4.2.0 software.Differentially expressed efferocytosis-related genes(DEERGs)were obtained,and KEGG and GO enrichment analyses were performed for DEERGs.The network analysis of protein-to-protein interactions(PPI)and identification of key genes of DEERGs and mitochondrial autophagy(PINK1,P62)were conducted.The key genes were imported into Coremine database for chinese herbal medicine prediction.Combined with the Traditional Chinese Medicine pathogenesis of myocardial fibrosis and the previous research progress,the highest frequency of Astragali Radix,Carthami Flos and Lepidii Semen was selected for animal experiments.Masson staining was used to analyze the degree of fibrosis.Mitochondrial swelling assay and mitochondrial membrane potential detection were performed to evaluate the extent of mitochondrial dysfunction.RT-PCR and Western blot were employed to measure the mRNA and protein expression of mitophagy-related genes to assess mitophagy levels.Cardiac-resident macrophages were isolated,treated with AngⅡ,and subsequently assessed for their efferocytosis of apoptotic endothelial cells.Results There were 428 DEGs between in the control group and model group,including 165 up-regulated and 263 down-regulated DEGs.250 ERGs were identified from GeneCards database,and 12 DEERGs were obtained by drawing the intersection of DEGs and ERGs.KEGG and GO enrichment analysis of DEERGs showed that the process involved cellular burial,mitochondrial autophagy and endothelial cell regulation.PPI network analysis and identification of key genes of DEERGs and mitochondrial autophagy(PINK1,P62)were performed.The top 10 key genes were HIF1A,PINK1,GABARAP,ARG1,PPARG,UCP2,SQSTM1,CD36,SIRT1 and RAB7A.The key genes were imported into Coremine database to search for corresponding chinese herbal medicine.The most frequent ones were Astragali Radix,Carthami Flos and Lepidii Semen.Experimental validation demonstrated that the Astragali Radix,Carthami Flos and Lepidii Semen formulation inhibited AngⅡ-induced myocardial fibrosis and improved cardiac function in rats.Notably,it significantly enhanced the efferocytosis of apoptotic endothelial cells by AngⅡ-treated resident macrophages,with the high-dose group showing the most pronounced effect(P<0.05).Mechanistic studies using the high-dose Astragali Radix,Carthami Flos and Lepidii Semen formulation group revealed that,compared to control and model groups,the treatment group exhibited reduced mitophagy and ameliorated mitochondrial dysfunction(P<0.05).Conclusion The Astragali Radix,Carthami Flos and Lepidii Semen formulation may inhibit excessive mitophagy by regulating the expression of PINK1 and P62,promote macrophage efferocytosis,and thereby suppress the formation of myocardial fibrosis.