[摘 要] 目的：探讨miR-433对结直肠癌（CRC）细胞干细胞特性及5-氟尿嘧啶（5-FU）耐药性的影响，并阐明其潜在的分子调控机制。方法： 建立5-FU耐药的HCT-116细胞株。采用RT-qPCR和WB法检测miR-433及干细胞特性标志物（SOX2、OCT4、Nanog）的表达。通过双萤光素酶报告基因实验、免疫荧光和核质分离实验验证miR-433与靶基因YAP1的关系及YAP1对其亚细胞定位的影响。通过拯救实验确认miR-433/YAP1轴的功能。利用CCK-8法检测细胞对5-FU的敏感性。结果： 与亲代HCT-116/P细胞相比，耐药的HCT-116细胞株中miR-433表达显著下调（P < 0.05），而干细胞特性标志物（SOX2、OCT4、Nanog）表达上调（P < 0.05）。过表达miR-433能够抑制干细胞标志物的表达，并直接靶向YAP1，阻碍其核定位（P < 0.05）。恢复YAP1的表达能够部分逆转miR-433对干细胞标志物的抑制作用（P < 0.05）。功能上，上调miR-433显著增强了CRC细胞对5-FU的敏感性，而此效应同样可被YAP1的重新表达所削弱（P < 0.05）。结论： miR-433通过直接靶向YAP1通路，抑制CRC细胞的干细胞特性相关分子表型，并增强其对化疗药物的敏感性。miR-433/YAP1轴可能成为克服CRC 5-FU耐药的潜在新靶点。

ObjectiveThis paper aims to investigate and evaluate the staged efficacy and safety of the representative empirical prescription of the “Zhu Ke Jiao” theory， Qijia Rougan prescription， combined with entecavir in the treatment of hepatic fibrosis in chronic hepatitis B. MethodsA multicenter randomized controlled clinical study was conducted， and 101 patients diagnosed with chronic hepatitis B-related hepatic fibrosis （CHB-HF） who met the diagnosis and inclusion criteria were randomly assigned to an observation group （Qijia Rougan prescription + entecavir） and a control group （entecavir）. The treatment duration was 24 weeks. Liver stiffness measurement （LSM）， fibrosis-4 index （FIB-4）， portal vein diameter， hepatitis B serology， biochemical indicators， hepatic fibrosis markers in serum ［hyaluronic acid （HA）， laminin （LN）， procollagen Ⅲ peptide （PⅢP）， and type Ⅳ collagen （Ⅳ-C）］， and traditional Chinese medicine syndrome scores were used as efficacy evaluation indicators. Efficacy assessments and explorations of different staged subgroups of Qijia Rougan prescription were conducted according to LSM values based on the Metavir pathological staging standard. ResultsA total of 98 cases were included for statistical analysis， with 49 cases in the observation group and 49 in the control group. The general data of the patients in both groups were comparable. Compared with the same group before treatment， the observation group showed a significant reduction in LSM and FIB-4 （P<0.01）， as well as notable improvements in LN， Ⅳ-C， and various TCM syndrome scores （P<0.05， P<0.01）. When compared to the control group after treatment， the observation group demonstrated significant improvements in LSM， FIB-4， and various TCM syndrome score indicators （P<0.05， P<0.01）， indicating that the observation group performed better than the control group. Subgroup analysis of the regression of hepatic fibrosis stages showed that compared to the same group before treatment， the observation group had better improvement in regression of stages F2 and F3 （P<0.05）. When compared to the control group after treatment， the observation group exhibited superior improvement in regression of stage F3 （P<0.05）. No adverse events occurred in either group during the treatment period. ConclusionCompared with entecavir alone， the combination of Qijia Rougan prescription and entecavir significantly improves the degree of hepatic fibrosis and clinical TCM symptoms in patients. The optimal intervention period is primarily during stage F3， which is a potential “interception” point of the “Zhu Ke Jiao” theory.

Objective: To systematically analyze the confirmatory positivity of different combinations of HBsAg screening results in blood testing, providing data to support the optimization of blood donor eligibility management. Methods: A retrospective analysis was conducted on blood screening data from 174 266 voluntary blood donor samples at the Chongqing Blood Center between October 2021 and September 2022. Samples with inconsistent results between the two HBsAg enzymelinked immunosorbent assays (ELISA) and individual donor nucleic acid testing (NAT) were confirmed using an electrochemiluminescence immunoassay (ECLIA) and a neutralization test. The detection efficacy of four different HBsAg ELISA reagents was compared using the HBsAg-confirmed positive samples. Results: A total of 767(0.44%) HBV-reactive (HB-sAg and/or HBV DNA reactive) samples were detected. Among them, 344 samples with discordant serological and NAT results were collected, of which 64(18.6%) were confirmed positive by neutralization test. Additionally, 5 samples that were neutralization-negative but double-reactive for HBsAg and HBV DNA were confirmed as positive according to FDA guidance, resulting in a total of 69(20.1%) confirmed HBsAg-positive samples. There were significant differences in the neutralization test confirmation rates among different screening result categories (P<0.05): The group with dual HBsAg reagent reactivity (double reactive) & NAT-negative had the highest confirmation rate (96.9%, 31/32); the group reactive to only reagent 2 (single reactive) had a rate of 25.7% (29/113); while the confirmation rates for samples reactive to only reagent 1 and samples with isolated HBV DNA positivity were extremely low [0(0/34) and 2.4%(4/165), respectively]. The four commercial reagents showed significant differences in their ability to detect confirmed positive samples that were initially single reactive (P<0.05). Conclusion: Given the performance variations among HBsAg screening reagents, thorough performance verification is essential before implementation. When NAT is negative, dual HBsAg reactivity in screening can serve as a basis for confirming infection and directly deferring blood donors. However, confirming infection in donors with single HBsAg reactivity is more challenging, necessitating supplementary tests to rule out infection risk.

Lung transplantation is a definitive treatment for end-stage lung disease and can significantly improve patient prognosis. However, postoperative complications such as infection, rejection, ischemia-reperfusion injury and chronic lung allograft dysfunction intertwine to form a complex pathological network, posing persistent challenges to long-term patient survival. In 2025, research teams worldwide have made systematic progress in the field of basic lung transplantation research. By integrating cutting-edge technologies including single-cell multi-omics, spatial transcriptomics and novel animal models, significant breakthroughs have been achieved in the evolutionary dynamics of drug-resistant infections, molecular mechanisms of immune regulation, programmed cell death, optimization of donor lung protection strategies and early warning of chronic lung allograft dysfunction. This article systematically reviews the representative advances in basic lung transplantation research worldwide in 2025, and deeply analyzes the implications of mechanistic breakthroughs for optimizing diagnosis and treatment strategies, aiming to anchor the direction for innovative breakthroughs and clinical translation in basic lung transplantation research.

Objective To explore the association between walking exercise guided by 6 minute walking test(6MWT)and the incidences of 3-year major adverse cardiovascular event(MACE)in elderly patients with acute coronary syndrome(ACS)after percutaneous coronary intervention(PCI).Methods A total of 628 elderly ACS patients who undergoing PCI and obtaining success-ful coronary revascularization in our department from November 2018 to April 2019 were enrolled,and divided into 6MWT group(n=147)and control group(n=481)based on participa-ting in walking exercise guided by 6MWT or not.All of them were followed up for 3 years.The incidences of MACE[including coronary target vascular restenosis,acute myocardial infarction,heart failure,ischemic or hemorrhagic stroke]and all-cause death were observed.Univariate and multivariate Cox proportional analyses and Kaplan-Meier survival curve analysis were employed for data statistical analyses.Results At the end of follow-up,the incidences of target vascular restenosis(6.9％vs 2.0％,P=0.028),heart failure(3.7％vs 0％,P=0.036),stroke(3.7％vs 0％,P=0.036),and total MACE incidence(15.0％vs 4.1％,P=0.000)were statistically higher in the control group than the 6MWT group.Kaplan-Meier survival curve analysis showed that the cumulative incidence of MACE was significantly lower in the 6MWT group than the control group(Plog rank=0.001).Multivariate Cox regression analysis showed that not participating in walking exercise guided by 6MWT was an independent risk factor for occurrence of 3-year MACE(HR=3.102,95％CI:1.327-7.250,P=0.009).Conclusion Walking exercise guided by 6MWT reduces the incidence of 3-year MACE and improves the long-term prognosis of elderly ACS patients after PCI.

OBJECTIVE Hospital-acquired infections have emerged as an increasingly prominent and hard-to-com-pletely-avoid problem,with their complexity and challenges continuing to intensify.As a major public health con-cern,these hospital-acquired infections pose a serious threat to the safety of individuals within hospitals.Among the various routes of transmission,airborne transmission is one of the most important pathways leading to hospi-tal-acquired infections.A variety pathogenic viruses can attach to infectious respiratory particles produced by hu-man body and spread through these particles.Patients in hospitals,as the primary group releasing respiratory in-fectious particles,have behaviors(such as breathing,coughing,talking,etc.)that are closely related to the trans-mission,dissemination and spread of pathogenic microorganisms.It is generally believed that pathogens re-leased into the air by patients will propagate and spread with air currents,thereby elevating the risk of infection.In order to comprehensively safeguard the safety of healthcare workers and patients,and effectively curb the occur-rence of hospital-acquired infections,it is essential to investigate the impact of pathogen-releasing behaviors on the transmission of pathogenic microorganisms.This paper aims to review the research progress on the impact of pathogen-releasing behaviors of patients on the transmission of respiratory pathogenic microorganisms within healthcare buildings,as well as to provide an outlook for further research directions.

A recombinant adenovirus(rAd)for expression of Mycobacterium tuberculosis(M.tb)c-di-AMP phosphodiesterase CnpB was constructed,and its induced humoral immune response was detected.The codon-optimized gene of M.tb CnpB was cloned into the adenoviral plasmid pcADV.The recombinant plasmid pcADV-CnpB was transfected into HEK293T cells,and expression was detected with Western blot.The recombinant plasmid pcADV-CnpB and the backbone plasmid were co-transfected into HEK293T cells to obtain the recombinant adenovirus rAd-CnpB.rAd-CnpB was amplified in HEK293T cells,and the target protein expression of rAd-CnpB was detected with Western blot and immunofluorescence.Mice were immunized with rAd-CnpB intranasally,and their sera and bronchoalveolar lavage fluid(BALF)were collected.ELISA was used to detect levels of antigen-specific antibodies.Restriction enzyme digestion and sequencing indicated that the recombinant plasmid pcADV-CnpB was successfully constructed and led to protein expression in eukaryotic cells.rAd-CnpB was packaged and produced in HEK293T cells.After amplification and purification,rAd-CnpB with a titer of 5.53×1010 PFU/mL was obtained.rAd-CnpB led to CnpB expression in HEK293T cells.Intranasal immunization with rAd-CnpB increased levels of IgG and secretory IgA in BALF and led to high levels of IgG in sera.rAd-CnpB,the recombinant adenovirus for expression of c-di-AMP phosphodiesterase CnpB was successfully constructed,and was found to induce antigen-specific humoral and mucosal immune responses through mucosal immunization.Thus,rAd-CnpB may be used in further research on new TB vaccine strategies.

Objective:In alignment with the practical teaching objectives of Diseases of the Locomotor System and the competency requirements for graduates of military medical universities, this study conducted a multidimensional modification of the traditional objective structured clinical examination (OSCE) to provide a novel training mode that enhances the effectiveness of practical teaching of this course.Methods:The traditional OSCE was modified from the four dimensions of integration, individuality, immersion, and intelligence. The first three stations were set as "diagnosis" and the last three stations were set as "operation" to reflect the "integration" of diagnosis and treatment. The last station of "operation" was set as "medical cares of combat and training injuries" to reflect "individuality" of military medical training. The method of situation creation was used to reflect "immersion", and the application of intelligent terminals reflected "intelligence". The study involved 50 trainees who were assigned to either a modified OSCE group or a traditional teaching group through a random drawing process. The teaching effectiveness was assessed through evaluation of theoretical knowledge, practical skills, and participant satisfaction. The statistical analysis was conducted by SPSS 22.0, with parametric data assessed by t-tests and non-parametric data assessed by chi-square tests. Results:Before entering the department, there were no statistically significant differences in theoretical ( P=0.832) and practical ( P=0.513) scores between the two groups of trainees. However, after the internship, the modified OSCE group demonstrated significantly enhanced scores compared to the traditional teaching group, both in theory assessment [(93.88±1.92) vs. (90.76±2.85), P=0.001] and skill assessment [(94.32±1.25) vs. (91.68±2.82), P=0.001]. Additionally, the self-assessment of clinical capability improvement by the modified OSCE group was markedly higher across all dimensions than the traditional teaching group ( P=0.001). Furthermore, the evaluation conducted by basic combat unit on the job competency of graduated trainees indicated that the modified OSCE group outperformed the traditional teaching group in basic clinical diagnosis and treatment [(4.72±0.46) vs. (3.44±0.71), P=0.001], emergency management of combat and training injuries [(4.72±0.46) vs. (3.52±0.71), P=0.001], application of information technology [(4.44±0.71) vs. (3.91±0.80), P=0.029], basic military qualities [(4.40±0.71) vs. (3.92±0.91), P=0.043], mental health and resilience [(4.36±0.70) vs. (3.68±0.85), P=0.003], and the capacity for continuous learning [(4.64±0.70) vs. (3.83±0.76), P=0.001]. Conclusions:The modified OSCE teaching mode can better meet the practical teaching requirements for Diseases of the Locomotor System in the military medical university and thus holds promise for further application.

On the occasion of the 110th Anniversary of the establishment of the Chinese Medical Association, the third conference of the Fourth Editorial Board of Chinese Journal of Digestive Surgery, and the Second Elite Group of Chinese Journal of Digestive Surgery, was successfully held in Kunming on July 4, 2025. This conference systematically summarizes the development experience of the journal over the past 20 years from three aspects: the role of ecological construction of thought in the discipline construction of digestive surgery, the display of the latest academic achievements in the field of digestive surgery, the development difficulties, and breakthrough paths of the discipline, and strategically plans the path of discipline construction in the new era.

Objective To investigate the effects of meridian and tendon acupuncture on the mitochondrial apoptosis of quadriceps femoris muscle cells in model rats of knee osteoarthritis(KOA);To explore its mechanism for the treatment of KOA.Methods Totally 40 SPF healthy male Wistar rats were randomly divided into sham-operation group(10 rats)and modeling group(30 rats).The modified Hulth method was used to establish a KOA rat model.The model rats were randomly divided into model group,celecoxib group and meridian and tendon acupuncture group,with 9 rats in each group.Corresponding intervention measures were given to each group for 14 consecutive days.Improved Lequesne MG score was used for evaluating knee joint function in rats,measuring thigh circumference,quadriceps femoris wet weight,wet weight maintenance rate and wet weight ratio of the affected limb,HE staining was used to observe the morphology of quadriceps femoris tissue,TUNEL method was used to detect apoptosis of quadriceps femoris cells,immunofluorescence was used to detect the expressions of reactive oxygen species(ROS),superoxide dismutase(SOD),Caspase-3 and Caspase-9 in quadriceps femoris muscle tissue,fluorescence double staining method was used to detect the co-expression of ROS and Caspase-3 in quadriceps femoris tissue,Western blot was used to detect the expressions of apoptosis related proteins in quadriceps femoris tissue.Results Compared with the sham-operation group,the improved Lequesne MG score of the knee joint in the model group rats increased(P<0.05),the circumference of the thigh,wet weight of the quadriceps femoris,wet weight maintenance rate and wet weight ratio of the affected limb decreased(P<0.05),the arrangement of quadriceps femoris muscle fibers was disordered and loose,with some muscle fibers dissolved and necrotic,accompanied by a large amount of inflammatory exudate,an increase in lymphocytes,and an increase in cell apoptosis index(P<0.05),the expressions of ROS,Caspase-3 and Caspase-9 in quadriceps femoris tissue increased,while the expression of SOD decreased(P<0.05),the protein expressions of Bcl-2,Bcl-XL,Bax,Cytochrome C(CytC),Caspase-3 and Caspase-9 in quadriceps femoris tissue increased(P<0.05).Compared with the model group,the improved Lequesne MG scores of the knee joint in the celecoxib group and the meridian and tendon acupuncture group decreased(P<0.05),while thigh circumference,quadriceps femoris wet weight,wet weight maintenance rate,wet weight ratio increased(P<0.05),the structure of quadriceps femoris muscle fibers was normal,the muscle membrane was relatively intact,the apoptosis index decreased(P<0.05),the expressions of ROS,Caspase-3 and Caspase-9 in quadriceps femoris tissue decreased,while the expression of SOD increased(P<0.05),the protein expressions of Bcl-2,Bcl-XL,Bax,CytC,Caspase-3 and Caspase-9 in quadriceps femoris muscle tissue decreased(P<0.05),and the expression trend of ROS and Caspase-3 in fluorescent double staining was consistent.Conclusion Meridian and tendon acupuncture can reduce ROS in the quadriceps femoris tissue of KOA model rats,inhibit the expressions of mitochondrial apoptosis-related proteins,thereby improving skeletal muscle strength,and play a certain therapeutic role.