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 ocular surface must maintain homeostasis to withstand continuous environmental stimuli, while increased tear evaporation represents a critical initiating event in the progression of evaporative dry eye disease. The composition, distribution, and function of the tear film lipid layer determine its anti-evaporative capacity. Multiple factors, including meibomian gland dysfunction, abnormalities of the lipid layer, blinking disorders, mucin deficiency, and epithelial barrier impairment, collectively weaken the evaporative barrier, subsequently inducing hyperosmotic stress and ocular surface inflammation, activating neuro-immune pathways, and thereby establishing a self-perpetuating cycle and contributing to disease chronicity. The environmental characteristics of northwestern regions, including low humidity, limited rainfall,strong winds, frequent windblown dust, and high evaporation rates, may further exacerbate these processes. This review integrates modern ocular surface pathological mechanisms with the theory of “Northwest dryness syndrome” to summarize the pathogenesis of evaporative dry eye disease and the effects of dry environmental exposure, aiming to provide a reference for risk assessment and intervention strategies for dry eye in Northwest China.

ObjectiveTo generate cynomolgus and rhesus monkey models carrying the Booroola fecundity (FecB) variant of bone morphogenetic protein receptor type 1B (BMPR1B) gene (designated BMPR1B^FecB) using adenine base edit technology and to explore genetic improvement strategies for enhancing the reproductive performance of laboratory macaques. MethodsFour single-guide RNAs (sgRNAs) were designed to target the BMPR1B c.746A>G locus in cynomolgus and rhesus monkeys. The sgRNAs were combined with the ABE8e-SpRY base editor, and the resulting editing complexes were microinjected into zygotes of both macaque species. Following in vitro culture of the injected embryos, nested PCR amplification was performed to detect the base-editing efficiency at the target site to screen for highly efficient sgRNAs. Subsequently, normally developing gene-edited embryos were transferred into the oviducts of surrogates. Seven days after birth, ear skin or peripheral blood samples were collected from the newborn monkeys for genotyping and mosaicism analysis, and the growth, development, and health status of the gene-edited monkeys were monitored. ResultsAmong the four candidate sgRNAs, sgRNA4 was identified as achieving 100% editing efficiency at the c.746A>G site. A total of 17 live monkey offspring were obtained via embryo transfer, including 8 cynomolgus monkeys and 9 rhesus monkeys. Genotyping results showed that all 8 newborn cynomolgus monkeys carried the c.746A>G point mutation, with an editing efficiency of 100% (8/8), of which 37.50% (3/8) were mosaics. Among the 9 newborn rhesus monkeys, 8 individuals carried the c.746A>G mutation, with an editing efficiency of 88.89% (8/9), of which 75.00% (6/8) were mosaics. ConclusionThe ABE8e-SpRY base-editing system enables precise editing at the c.746A>G (p.Q249R) locus of the BMPR1B gene in both cynomolgus and rhesus monkey embryos, resulting in the successful establishment of laboratory monkey models carrying the BMPR1B^FecB variant. These models provide a valuable foundation for future studies of the regulatory role of the BMPR1B^FecB variant in reproductive performance at the non‑human primate level, as well as for exploring genetic improvement strategies to enhance reproductive performance in laboratory monkeys.

ObjectiveTo verify the protective effect and pharmacological mechanism of Qishen granules against doxorubicin-induced cardiotoxicity， thus providing a basis for developing new drug options for clinical treatment of cardio-oncology disease. MethodsFifty mice were randomly assigned into a normal group （n=10） and a modeling group （n=40）. The model was established by intravenous injection of doxorubicin （5 mg·kg^-1） via the tail vein， once a week for 4 consecutive weeks. The normal group was injected with an equal volume of normal saline during the same period. The modeled mice were further assigned into four groups： doxorubicin （no drug control）， low-dose （3.33 mg·kg^-1） Qishen granules， high-dose （6.67 mg·kg^-1） Qishen granules， and enalapril （15 mg·kg^-1）. Mice were administrated with corresponding drugs by gavage for 4 consecutive weeks. At the end of the experiment， echocardiography， histopathological staining， and live-cell imaging were employed to determine relevant indicators， on the basis of which the protective effect of Qishen granules against doxorubicin-induced cardiotoxicity was evaluated. ResultsCompared with the normal group， doxorubicin treatment reduced the left ventricular ejection fraction （LVEF） （P<0.05） and caused pathological changes including disarrangement of cardiomyocytes， increased cell spacing， cardiomyocyte hypertrophy， and increased apoptosis in the heart tissue. Meanwhile， doxorubicin prolonged the mitochondria-lysosome contact time， downregulated the expression of TBC1 domain family member 15 （TBC1D15） （P<0.01）， upregulated the levels of microtubule-associated protein 1 light chain 3 （LC3） and sequestosome 1 （p62） （P<0.01）， and induced mitochondrial cristae breakage， resulting in mitochondrial dysfunction. Compared with the doxorubicin group， intervention with Qishen granules reversed the above indicators. It not only increased the LVEF （P<0.01） and alleviated pathological damage to the heart， but also upregulated the expression of TBC1D15 （P<0.01）， shortened the mitochondria-lysosome contact time （P<0.05）， and downregulated the protein levels of LC3 and p62. When TBC1D15 in cells was knocked down， the autophagy-activating and mitochondrion-protecting effects of Qishen granules partially disappeared. ConclusionQishen granules upregulate the expression of TBC1D15 to shorten the mitochondria-lysosome contact time， thereby activating autophagy and ultimately exerting a protective effect against doxorubicin-induced cardiotoxicity.

ObjectiveTo verify the protective effect and pharmacological mechanism of Qishen granules against doxorubicin-induced cardiotoxicity， thus providing a basis for developing new drug options for clinical treatment of cardio-oncology disease. MethodsFifty mice were randomly assigned into a normal group （n=10） and a modeling group （n=40）. The model was established by intravenous injection of doxorubicin （5 mg·kg^-1） via the tail vein， once a week for 4 consecutive weeks. The normal group was injected with an equal volume of normal saline during the same period. The modeled mice were further assigned into four groups： doxorubicin （no drug control）， low-dose （3.33 mg·kg^-1） Qishen granules， high-dose （6.67 mg·kg^-1） Qishen granules， and enalapril （15 mg·kg^-1）. Mice were administrated with corresponding drugs by gavage for 4 consecutive weeks. At the end of the experiment， echocardiography， histopathological staining， and live-cell imaging were employed to determine relevant indicators， on the basis of which the protective effect of Qishen granules against doxorubicin-induced cardiotoxicity was evaluated. ResultsCompared with the normal group， doxorubicin treatment reduced the left ventricular ejection fraction （LVEF） （P<0.05） and caused pathological changes including disarrangement of cardiomyocytes， increased cell spacing， cardiomyocyte hypertrophy， and increased apoptosis in the heart tissue. Meanwhile， doxorubicin prolonged the mitochondria-lysosome contact time， downregulated the expression of TBC1 domain family member 15 （TBC1D15） （P<0.01）， upregulated the levels of microtubule-associated protein 1 light chain 3 （LC3） and sequestosome 1 （p62） （P<0.01）， and induced mitochondrial cristae breakage， resulting in mitochondrial dysfunction. Compared with the doxorubicin group， intervention with Qishen granules reversed the above indicators. It not only increased the LVEF （P<0.01） and alleviated pathological damage to the heart， but also upregulated the expression of TBC1D15 （P<0.01）， shortened the mitochondria-lysosome contact time （P<0.05）， and downregulated the protein levels of LC3 and p62. When TBC1D15 in cells was knocked down， the autophagy-activating and mitochondrion-protecting effects of Qishen granules partially disappeared. ConclusionQishen granules upregulate the expression of TBC1D15 to shorten the mitochondria-lysosome contact time， thereby activating autophagy and ultimately exerting a protective effect against doxorubicin-induced cardiotoxicity.

Objective: To explore the therapeutic effects of different surgical methods for temporomandibular joint disc reduction and anchoring surgery, providing reference for optimizing this surgical procedure. Method: The study was approved by the hospital ethics committee. 173 patients (195 joints) who underwent temporomandibular joint disc repositioning and anchoring surgery were selected for retrospective analysis. Patients were categorized into groups A (traditional preauricular incision-scalpel/tissue scissors anterior attachment release), 35 patients (40 joints), B (traditional preauricular incision-plasma bipolar radiofrequency electrode anterior attachment release), 42 patients (46 joints), C (revised tragus incision - scalpel/tissue scissors anterior attachment release), 50 patients (58 joints), and D (revised tragus incision-plasma bipolar radiofrequency electrode anterior attachment release), 46 patients (51 joints). After a 6-month postoperative follow-up, the differences in maximum mouth opening (MMO), visual analogue scale (VAS), effective rate of joint disc reduction, incidence of preauricular numbness, obvious scars among patients in each group at 1, 3, and 6 months were compared postoperatively. Results: After surgery, the MMO of all four groups of patients initially shrunk and then gradually increased compared to before surgery. At the 1-month follow-up after surgery, the plasma bipolar radiofrequency release (B+D) group had a smaller impact on the patient’s MMO compared to the surgical knife/tissue scissors release (A+C) group (P < 0.05). Postoperative VAS scores for all four groups showed a gradual decrease from pre-operative levels, with the (B+D) group scoring significantly lower in the first month post-surgery compared to the (A+C) group (P < 0.05). Six months post-surgery, the rate of joint disc reduction of the four groups were higher than 95%, with no significant differences observed between the groups (P > 0.05). Patients in the revised tragus incision (C+D) group experienced a lower rate of preauricular numbness compared to those in the traditional preauricular incision (A+B) group (4.59% vs. 12.79%, P < 0.05), The incidence of obvious scars in the (C+D) group was significantly lower than that in the (A+B) group (3.67% vs. 23.26%, P < 0.05). Conclusion The revised tragus incision is superior to traditional preauricular incision in terms of protecting the auriculotemporal nerve and the scars were more inconspicuous. Further, the plasma bipolar radiofrequency electrode is superior to the scalpel/tissue scissors in terms of mouth opening recovery and pain control. For temporomandibular joint disc reduction and anchoring surgery, a modified tragus incision combined with plasma bipolar radiofrequency electrode to release the anterior attachment of the joint disc can be recommended as a surgical option.

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.

ObjectiveTo investigate the difference in the risk of cardiovascular diseases between patients with different subtypes of non-alcoholic fatty liver disease （NAFLD） from the perspective of metabolism， since cardiovascular events induced by metabolic disorders are the leading cause of death in NAFLD. MethodsThe cluster sampling method was used to conduct a multicenter cross-sectional study among three representative hospitals in Pudong New Area of Shanghai， China. A total of 37 122 sets of physical examination data from July 2022 to June 2023 were collected and stratified according to body mass index （BMI）. The chi-square test was used for comparison of continuous data between groups， and a multivariable Logistic regression analysis was used to investigate the association between NAFLD subtypes and cardiometabolic risk factors. ResultsA total of 9 372 cases of NAFLD were detected， with a detection rate of 25.25%， and more than 97% of these patients were diagnosed with metabolic associated fatty liver disease （MAFLD）. The subgroup analysis showed that the detection rates of lean， overweight， and obese NAFLD were 7.72%， 33.99%， and 63.56%， respectively. Compared with the patients with lean or overweight NAFLD， the patients with obese NAFLD showed a significantly higher proportion of patients with abnormalities in blood pressure， blood glucose， triglyceride （TG）， high-density lipoprotein （HDL） or uric acid （all P<0.001）. Among related risk factors， lean NAFLD was associated with the increase in total cholesterol （TC）（P<0.05）， while overweight NAFLD and obese NAFLD were not associated with TC abnormalities （P>0.05）； obese NAFLD was not associated with TG abnormalities， while lean NAFLD and overweight NAFLD were associated with TG abnormalities （both P<0.05）； all types of NAFLD were associated with the abnormalities of waist-hip ratio， blood pressure， blood glucose， low-density lipoprotein， HDL， and uric acid （all P<0.05）. ConclusionThe detection rates of different subtypes of NAFLD in Shanghai Pudong are close to those reported in China and globally， and the epidemiologic data of NAFLD can be used analogously for MAFLD. There are certain differences in the distribution and association of cardiometabolic risk factors between different subtypes of NAFLD， and targeted interventions should be formulated based on the metabolic characteristics of each type of NAFLD.