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.

[摘 要] 目的：探讨异位表达血红蛋白亚基（HBA/HBB）对缺氧条件下嵌合抗原受体T细胞（CAR-T细胞）功能障碍的改善作用及其对肿瘤细胞的杀伤效应。方法：全基因合成技术合成靶向HER2的CAR序列，构建共表达HBA或HBB的CAR慢病毒载体，包装慢病毒后感染人原代T淋巴细胞，制备异位表达HBA/HBB的CAR-T细胞，命名为HBA CAR-T和HBB CAR-T。采用缺氧探针检测小鼠实体瘤缺氧状态。通过流式细胞术检测瘤内CAR-T细胞占比、异位表达血红蛋白亚基的CAR-T细胞阳性率及CAR-T细胞的活性氧、凋亡水平。WB法检测HBA CAR-T和HBB CAR-T内相关血红蛋白亚基表达情况，采用细胞计数板计数检测细胞增殖水平，通过萤光素酶报告基因法检测CAR-T细胞对肿瘤细胞的杀伤能力，qPCR检测CAR-T细胞中缺氧诱导因子-1α（HIF-1α）表达水平，利用MitoXpress Intra试剂盒检测CAR-T细胞内氧气含量。结果：不同细胞构建的实体瘤模型均存在明显缺氧情况，且CAR-T细胞浸润水平与缺氧程度呈显著负相关（P < 0.000 1）。HBA CAR-T与HBB CAR-T构建成功（阳性率 > 60%），相应血红蛋白亚基可稳定表达。缺氧环境下HBA CAR-T和HBB CAR-T的ROS水平、凋亡水平显著下降，增殖、对肿瘤细胞的体外杀伤能力显著强于传统CAR-T细胞（均P < 0.05）。HBA CAR-T与HBB CAR-T内HIF-1α表达降低（均P < 0.001），且缺氧程度显著降低（均P < 0.001）。结论：异位表达血红蛋白亚基可改善缺氧条件下CAR-T细胞功能障碍并增强其对肿瘤细胞的体外杀伤作用。

Childhood simple obesity has become a significant public health issue in China. Modern medicine primarily relies on lifestyle interventions and often suffers from poor long-term compliance， while pharmacological options are limited and associated with potential adverse effects. Traditional Chinese Medicine （TCM） has a long history in the prevention and management of this condition， demonstrating eight distinct advantages， including systematic theoretical foundation， diversified therapeutic approaches， definite therapeutic efficacy， high safety profile， good patient compliance， comprehensive intervention strategies， emphasis on prevention， and stepwise treatment protocols. Additionally， TCM is characterized by six distinctive features： the use of natural medicinal substances， non-invasive external therapies， integration of medicinal dietetics， simple exercise regimens， precise syndrome differentiation， and diverse dosage forms. By combining internal and external treatments， TCM facilitates individualized regimen adjustment and holistic regulation， demonstrating remarkable effects in improving obesity-related metabolic indicators， regulating constitutional imbalance， and promoting healthy behaviors. However， challenges remain， such as inconsistent operational standards， insufficient high-quality clinical evidence， and a gap between basic research and clinical application. Future efforts should focus on accelerating the standardization of TCM diagnosis and treatment， conducting multicenter randomized controlled trials， and fostering interdisciplinary integration， so as to enhance the scientific validity and international recognition of TCM in the prevention and treatment of childhood obesity.

AIM: To explore the application effect of eyelid margin incision combined with floating bridge technique in the treatment of ptosis patients, and to analyze its effects on the margin reflex distance 1(MRD1)and the aesthetic effect of the eyes. METHODS:Prospective study. This study enrolled patients with blepharoptosis treated at the Department of Ophthalmology of the hospital between January 2023 and July 2025, who were assigned to either an observation group or a control group. The patients in the control group received levator aponeurosis shortening surgery through conventional double-eyelid incision, while the patients in the observation group underwent levator aponeurosis shortening surgery using the eyelid margin incision combined with floating bridge technique. Subsequently, the surgical-related indicators,the correction effect of ptosis, the aesthetic effect, the ocular function indicators, the postoperative complication occurrence rate, and recurrence rate were compared and analyzed between two groups.RESULTS:A total of 100 patients(100 eyes)with blepharoptosis were enrolled in this study, with 50 patients(50 eyes)in the observation group and 50 patients(50 eyes)in the control group. The observation group had a mean age of 27.33±5.32 y, including 20 males and 30 females. The control group had a mean age of 27.64±4.66 y, including 24 males and 26 females. Compared with the control group, the operative time of the observation group was longer, the postoperative swelling resolution time and natural recovery time were shorter,and the correction rate was higher(P<0.05). Three months after the surgery, the VSS score and POSAS score of the observation group were lower than those of the control group, while the scores for eyelid curvature, symmetry of eyelid height, and the PFH, MRD1, palpebral fissure size, and upper eyelid levator muscle strength were all higher than those of the control group(all P<0.05). However, there was no significant difference in postoperative complication occurrence rate and recurrence rate between two groups(all P>0.05).CONCLUSION:The combined use of eyelid margin incision and floating bridge technique for treating ptosis results in slightly longer operation time, but it can shorten the postoperative recovery period, effectively ameliorate the MRD1 and ocular function of patients, and enhance the correction effect and aesthetic quality of the eyes. In addition, it has a low incidence of complications and guaranteed safety.

Objective To examine the changing prevalence and antimicrobial resistance profiles of Burkholderia cepacia in 52 hospitals across China from 2015 to 2021.Methods A total of 9 261 strains of B.cepacia were collected from 52 hospitals between January 1,2015 and December 31,2021.Antimicrobial susceptibility of the strains was tested using Kirby-Bauer method or automated antimicrobial susceptibility testing systems according to a unified protocol.The results were interpreted according to the breakpoints released in the Clinical & Laboratory Standards Institute(CLSI)guidelines(2023 edition).Results A total of 9 261 strains of B.cepacia were isolated from all age groups,especially elderly patients.The proportion was 11.1％(1 032 strains)in children,significantly lower than the proportion in adults.About half(46.5％,4 310/9 261)of the strains were isolated from patients at least 60 years old and 42.3％(3 919/9 261)of the strains were isolated from young adults.Most isolates(71.1％)were isolated from sputum and respiratory secretions,followed by urine(10.7％)and blood samples(8.1％).B.cepacia isolates were highly susceptible to the five antimicrobial agents recommended in the CLSI M100 document(33rd edition,2023).B.cepacia isolates showed relatively higher resistance rates to meropenem and levofloxacin.However,the resistance rates to ceftazidime,trimethoprim-sulfamethoxazole,and minocycline remained below 8.1％.The percentage of B.cepacia strains resistant to levofloxacin was the highest compared to other antibiotics in any of the three age groups(from 12.4％in the patients＜18 years old to 20.6％in the patients aged 60 years or older).Conclusions B.cepacia is one of the clinically important non-fermenting gram-negative bacteria.Accurate and timely reporting of antimicrobial susceptibility test results and ongoing antimicrobial resistance surveillance are helpful for rational prescription of antimicrobial agents and proper prevention and control of nosocomial infections.

Body weight abnormalities, including overweight, obesity, and underweight, have become a dual public health challenge in Chinese adults: overweight and obesity lead to a variety of chronic complications, while underweight increases the risks of malnutrition, sarcopenia, and organ dysfunction. To systematically address these issues, multidisciplinary experts in endocrinology, sports science, nutrition, and psychiatry from various regions have held multiple weight management seminars. Based on the latest epidemiological data and clinical evidence, they expanded the guideline to include assessment and intervention strategies for underweight, in addition to the core content of obesity management. This guideline outlines the etiological mechanisms, evaluation methods, and multidimensional management strategies for overweight and obesity, covering key areas such as diagnosis and assessment, medical nutrition therapy, exercise prescription, pharmacological intervention, and psychological support. It is intended to provide a scientific and standardized approach to weight management across the adult population, aiming to curb the rising prevalence of obesity, mitigate complications associated with abnormal body weight, and improve nutritional status and overall quality of life.

Intra-aortic balloon pump(IABP),as the most commonly used percutaneous mechanical circulatory assist device,is routinely implanted through the femoral artery pathway.However,when implantation through the femoral artery pathway is difficult or contraindicated,other pathways including the brachial artery pathway,axillary artery pathway,etc.can be considered.IABP assisted interventional treatment for complex high-risk and indicated patients(CHIP)can reduce the risk of intraoperative complications,stabilize hemodynamics,increase the complete revascularization rate of CHIP,and improve long-term prognosis.The reverse guidewire technique can improve the success rate of percutaneous coronary intervention(PCI)for chronic total occlusion(CTO)of coronary arteries.In clinical practice,the reverse pathway is often chosen in addition to the forward pathway,such as the contralateral radial and brachial artery pathway,femoral artery pathway,etc.This article reports a case of bilateral femoral artery occlusion,in which IABP was implanted through the left brachial artery pathway,and then with the assistance of IABP,the right coronary artery CTO lesion was successfully opened through the right radial artery in the forward direction and the right brachial artery in the reverse direction,with the aim of providing reference for clinical PCI treatment for such special cases.

Objective To summarize the changing prevalence of carbapenem resistance in Enterobacterales based on the data of CHINET Antimicrobial Resistance Surveillance Program from 2015 to 2021 for improving antimicrobial treatment in clinical practice.Methods Antimicrobial susceptibility testing was performed using a commercial automated susceptibility testing system according to the unified CHINET protocol.The results were interpreted according to the breakpoints of the Clinical & Laboratory Standards Institute(CLSI)M100 31st ed in 2021.Results Over the seven-year period(2015-2021),the overall prevalence of carbapenem-resistant Enterobacterales(CRE)was 9.43％(62 342/661 235).The prevalence of CRE strains in Klebsiella pneumoniae,Citrobacter freundii,and Enterobacter cloacae was 22.38％,9.73％,and 8.47％,respectively.The prevalence of CRE strains in Escherichia coli was 1.99％.A few CRE strains were also identified in Salmonella and Shigella.The CRE strains were mainly isolated from respiratory specimens(44.23±2.80)％,followed by blood(20.88±3.40)％and urine(18.40±3.45)％.Intensive care units(ICUs)were the major source of the CRE strains(27.43±5.20)％.CRE strains were resistant to all the β-lactam antibiotics tested and most non-β-lactam antimicrobial agents.The CRE strains were relatively susceptible to tigecycline and polymyxins with low resistance rates.Conclusions The prevalence of CRE strains was increasing from 2015 to 2021.CRE strains were highly resistant to most of the antibacterial drugs used in clinical practice.Clinicians should prescribe antimicrobial agents rationally.Hospitals should strengthen antibiotic stewardship in key clinical settings such as ICUs,and take effective infection control measures to curb CRE outbreak and epidemic in hospitals.

Objective To characterize the changing species distribution and antibiotic resistance profiles of respiratory isolates in hospitals participating in the CHINET Antimicrobial Resistance Surveillance Program from 2015 to 2021.Methods Commercial automated antimicrobial susceptibility testing systems and disk diffusion method were used to test the susceptibility of respiratory bacterial isolates to antimicrobial agents following the standardized technical protocol established by the CHINET program.Results A total of 589 746 respiratory isolates were collected from 2015 to 2021.Overall,82.6％of the isolates were Gram-negative bacteria and 17.4％were Gram-positive bacteria.The bacterial isolates from outpatients and inpatients accounted for(6.0±0.9)％and(94.0±0.1)％,respectively.The top microorganisms were Klebsiella spp.,Acinetobacter spp.,Pseudomonas aeruginosa,Staphylococcus aureus,Haemophilus spp.,Stenotrophomonas maltophilia,Escherichia coli,and Streptococcus pneumoniae.Each microorganism was isolated from significantly more males than from females(P＜0.05).The overall prevalence of methicillin-resistant S.aureus(MRSA)was 39.9％.The prevalence of penicillin-resistant S.pneumoniae was 1.4％.The prevalence of extended-spectrum β-lactamase(ESBL)-producing E.coli and K.pneumoniae was 67.8％and 41.3％,respectively.The overall prevalence of carbapenem-resistant E.coli,K.pneumoniae,Enterobacter cloacae,Pseudomonas aeruginosa,and Acinetobacter baumannii was 3.7％,20.8％,9.4％,29.8％,and 73.3％,respectively.The prevalence of β-lactamase was 96.1％in Moraxella catarrhalis and 60.0％in Haemophilus influenzae.The H.influenzae isolates from children(＜18 years)showed significantly higher resistance rates to β-lactam antibiotics than the isolates from adults(P＜0.05).Conclusions Gram-negative bacteria are still predominant in respiratory isolates associated with serious antibiotic resistance.Antimicrobial resistance surveillance should be strengthened in clinical practice to support accurate etiological diagnosis and appropriate antimicrobial therapy based on antimicrobial susceptibility testing results.

Objective To explore the therapeutic effect of picroside Ⅱ(P Ⅱ)on diabetes nephropathy(DN)rats by regulating fatty acid synthase(Fas)/fatty acid synthase ligand(FasL)signaling pathway.Methods A DN rat model was constructed by combining high sugar and high-fat diet with streptozotocin(STZ)injection.DN rats were grouped into model group(DN group),low,medium and high dose picroside Ⅱ groups(P-L group,P-M group,P-H group),and high dose picroside Ⅱ+Fas recombinant protein group(P-H+rh-Fas group),with healthy rats as control group,and 18 rats in each group.Fasting blood glucose(FBG),body mass,24-hour urinary protein(24h UTP),and renal function(SCr,BUN)were measured in rats.Hematoxylin-eosin(HE)staining and Masson staining were applied to observe the pathological changes in renal tissue of rats;immunohistochemistry and Western blot were applied to detect the expression of RAGE,COL-Ⅳ and pathway proteins,respectively.Results Compared with the control group,rats in DN group showed thickening of the glomerular basement membrane,mesangial proliferation,tubular degeneration,dilation,atrophy,fatty degeneration,obvious collagen deposition,higher levels of FBG,24h UTP,SCr,BUN,TNF-α,IL-6 and IL-1β,body mass loss,and higher expression of RAGE,COL-Ⅳ,Fas and FasL(P＜0.05).Compared with the DN group,the glomerular and tubular lesions were reduced and collagen deposition was decreased in the P-L,P-M and P-H groups,furthermore,the FBG,24h UTP,SCr,BUN,TNF-α,IL-6,IL-1β levels were lower,body mass was higher,and the RAGE,COL-Ⅳ,Fas,FasL expression was lower(P＜0.05).Compared with the P-H group,the renal tissue lesions in the P-H+rh-Fas group worsened,the FBG,24 h UTP,SCr,BUN,TNF-α,IL-6,IL-1β levels were higher,body mass was lower,and the RAGE,COL-Ⅳ,Fas,FasL expression was higher(P＜0.05).Conclusion Picroside Ⅱ exerts therapeutic effects on DN rats by inhibiting the Fas/FasL signaling pathway.