[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。

ObjectiveTo clarify the expression of TRIM28 in non-M3 acute myeloid leukemia （AML） and its correlation with clinical indicators and prognosis， and to further explore the effect of TRIM28 expression levels on the proliferation and apoptosis of AML cells using small interfering RNA. MethodsThe GSE34577 dataset was analyzed using R software to compare TRIM28 expression between healthy controls and non-M3 acute myeloid leukemia （AML） patients. Clinical samples from non-M3 AML patients were collected， with TRIM28 expression levels measured using real-time quantitative PCR （qPCR）. The analysis focused on correlations between TRIM28 expression and various clinical indicators， treatment efficacy， and patient prognosis. Furthermore， small interfering RNA （siRNA） technology was employed to downregulate TRIM28 expression in human primary AML cells （HL60 cell line）. The effects on cell proliferation and apoptosis were then assessed through CCK-8 assays and flow cytometry， respectively. ResultsThe results showed that TRIM28 was up-regulated in non-M3 AML of both online database GSE34577 and clinical samples （P<0.000 1）， TRIM28 expression of new diagnosis group and relapsed refractory group was higher than iron deficiency anemia group （P<0.01）， and there was no significance between different French-American-British classification systems subtype. TRIM28 expression was higher in non-M3 AML patients with a poor genetic prognosis stratified as moderate than in the good prognosis group， and TRIM28 expression was associated with NPM1 combined with the FLT3-ITD mutation， positively correlated with age， bone marrow blast， peripheral blood blast and white blood cell， negatively correlated with hemoglobin. In addition， interference TRIM28 greatly inhibited cell proliferation and promoted cell apoptosis. ConclusionThis study reveals that TRIM28 is highly expressed in non-M3 AML and associated with prognosis， and plays a key role in the proliferation and apoptosis of AML cells， suggesting that TRIM28 may serve as a novel therapeutic target for non-M3 AML.

Renal hilar tumors pose significant surgical challenges due to their proximity to major renal vessels and the collecting system. In robot-assisted partial nephrectomy, although both anterior and posterior lip tumors are considered renal hilar tumors, their operative difficulty and complication profiles differ markedly owing to distinct anatomical relationships and vascular adjacency. Anterior lip tumors, often involving tributaries of the renal vein, carry a relatively higher risk of intraoperative and postoperative bleeding. In contrast, posterior lip tumors, closer to the renal artery, are more likely to prolong warm ischemia time, thereby increasing the risk of postoperative renal impairment. Regarding urine leakage, although anterior lip tumors are theoretically closer to the collecting system and may pose a slightly elevated risk, precision reconstruction strategies—such as lip reconstruction based on sinus space anatomy—can effectively mitigate this risk. However, the specific difference in urine leakage risk between anterior and posterior lip tumors warrants further investigation. Both anterior and posterior lip tumors of the kidney can achieve low positive surgical margin rates in experienced centers. Complication risks are affected by multiple factors, including tumor anatomical complexity, warm ischemia time, and surgeon experience. Notably, R.E.N.A.L. score＞8 is associated with significantly increased complication risks. Preoperative meticulous imaging evaluation, selection of an appropriate surgical approach based on tumor location, controlled warm ischemia time, and application of delicate suturing and reconstruction techniques are essential strategies to reduce complications.

Objective To explore the application of cognitive interview in the development of Experience Scale of Patient Sense of Gain.Methods Purposive sampling method were used to select 13 inpatients of a tertiary grade A hospital in Xi'an,14 outpatients from a community health service center in Guangzhou,and 14 inpatients of a tertiary hospital in Guangzhong from February to April 2023 as the interviewees.The measurement tool was the preliminary version of Experience Scale of Patient Sense of Gain which was preliminary phase of research results of the group.Three rounds of cognitive interviews were conducted to explore the respondent'sunderstanding towards the items of the scale.The collected data were analyzed by thematic coding and the items of the scale were revised by the results of interviews and group discussion.Results After the first round of interviews,eight doubtful items were discussed and revised;after the second round of interviews,three doubtful entries were discussed and revised;and after the third round of interviews,all the interviewees were able to correctly understand the scale.Conclusion Cognitive interviews can provide insights into the target population's understanding of Experience Scale of Patient Sense of Gain,effectively addressing the problems of understanding differences between the developer and respondent in the process of the scale development,thus improving the accuracy of the scale.

Objective To analyze the willingness and influencing factors of doctors in Beijing public hospitals to set up clinics in the community,and to provide scientific basis for promoting quality health technicians to go down to the grassroots and realize hierarchical diagnosis and treatment.Methods Doctors from 6 specialized hospita ls and 3 general hospitals in Beijing were selected as subjects for questionnaire survey.By stratified sampling,a questionnaire survey was conducted on 518 doctors from 9 public hospitals in Beijing through stratified sampling method.Descriptive analysis,rank sum test and ordered logistic regression were used to explore doctors'willingness to open clinics and its influencing factors.Results In all survey respondents,212(40.93%)physicians were very or somewhat willing to open a clinic in the community,physicians with higher benefit perception,and greater willingness to publicize relevant policies were more willing to set up clinics.Conclusion Opening of clinics by doctors in public hospitals is affected by perceived benefits and willingness to advocate policies.It is suggested to further improve relevant systems and policies,reduce the risk of doctors opening clinics,and promote doctors in public hospitals to open clinics in communities,so as to realize the sinking of high-quality health human resources to the grassroots.

Objective To explore the constituent elements of patients'sense of gain experience and provide theoretical insights for improving patient satisfaction with healthcare services.Methods Purposive sampling and convenience sampling were employed to select 46 experts in health management from universities,hospitals,government agencies,and academic societies across China for semi-structured interviews.Qualitative analysis of the interview data was conducted using grounded theory.Results Through three-level coding,159 initial concepts,19 categories,7 main categories,and 2 core categories were identified.A conceptual framework for the constituent elements of patients'sense of gain experience was constructed,centered on two core dimensions:micro-level experience and macro-level acquisition.Conclusion The concept of"patients'sense of gain experience"enriches the theoretical framework of"sense of gain"and breaks through the unidimensional approach to evaluating medical service quality.Its enhancement requires collaborative efforts from multiple stakeholders,including medical institutions,governments,policymakers,and other actors.

OBJECTIVE To investigate the expression and clinical significance of microRNA-223(miR-223),mono-cyte chemoattractant protein-1(MCP-1),and basic fibroblast growth factor(bFGF)in patients with colostomy infection after rectal cancer surgery.METHODS One hundred patients with rectal cancer who underwent surgery in Zhejiang Provincial People's Hospital between Jun.2022 and Jun.2024 were chosen retrospectively and divided in-to an infection group(n=27)and a non-infection group(n=73)based on whether colostomy infection occurred within seven days after surgery.The etiological features of postoperative hospital-acquired infection were analyzed,and the differences in serum miR-223,bFGF,and MCP-1 levels between the infected and non-infected groups were detected.The receiver operating characteristic(ROC)curve was used to analyze the predictive values of ser-um miR-223,bFGF,and MCP-1 for postoperative colostomy infection.RESULTS A total of 30 pathogenic strains were isolated from 27 patients in the infection group,including 17 gram-negative bacteria(56.67%),11 gram-positive bacteria(36.67%),and 2 fungi(6.67%),with Escherichia coli being the most common.The serum lev-els of miR-223 and MCP-1 were higher in the infected group than those in the non-infected group,while bFGF was lower in the infected group(P<0.05).The ROC curve analysis showed that the area under the curve(AUC)for the combined detection of serum miR-223,bFGF,and MCP-1 was 0.944,with the sensitivity of 88.88%and spe-cificity of 94.52%.CONCLUSIONS Postoperative colostomy infection in rectal cancer patients is primarily caused by E.coli and is associated with changes in serum miR-223,bFGF and MCP-1 levels.Abnormally high expression of miR-223 and MCP-1 and abnormally low expression of bFGF can predict postoperative colostomy infection in rectal cancer,which can provide an important basis for clinical diagnosis of postoperative infections in the rectal cancer patients.

Objective To investigate the characteristics of intestinal flora and its correlation with peripheral blood microinflammatory factors in patients with pulmonary tuberculosis complicated with diabetes mellitus(PTB-DM).Methods A total of 162 patients with PTB-DM admitted to the hospital from September 2022 to September 2024 were selected as the PTB-DM group,and another 150 healthy subjects who underwent physi-cal examinations during the same period in the hospital were selected as the control group.The clinical data of all research subjects was collected.The composition of intestinal flora of the research subjects was analyzed by using 16S rRNA gene sequencing technology.The levels of peripheral blood microinflammatory factors[inter-feron-γ(IFN-γ),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)]in all research subjects were detected.The clinical data,composition of intestinal flora and levels of microinflammatory factors of the PTB-DM group were compared with those of the control group.Spearman correlation analysis was used to analyze the correlation between the relative abundance of intestinal flora and the levels of peripheral blood microin-flammatory factors in patients with PTB-DM.Results The relative abundances of Bacteroidales and Clostridi-ales in the PTB-DM group were significantly lower than those in the control group,while the relative abun-dances of Enterobacterales and Actinobacteriales were significantly higher than those in the control group,and the differences were statistically significant(P<0.05).The levels of IFN-γ,IL-6 and TNF-α in the PTB-DM group were significantly higher than those in the control group,and the differences were statistically signifi-cant(P<0.05).The relative abundances of Bacteroidales and Clostridiales in the PTB-DM group were nega-tively correlated with the levels of IFN-γ,IL-6 and TNF-α.The relative abundances of Enterobacterales and Actinobacteriales were positively correlated with the levels of IFN-γ,IL-6 and TNF-α(all P<0.05).Conclu-sion There is a significant imbalance of intestinal flora in patients with PTB-DM,and the levels of microin-flammatory factors IFN-γ,IL-6 and TNF-α in peripheral blood are significantly increased,and are closely re-lated to the relative abundance of specific flora.

Objective:To develop and validate machine learning models for predicting the cumulative live birth rate (CLBR) following in vitro fertilization (IVF) and to analyze key predictive features using SHAP values. Methods:This retrospective study included data from patients who underwent IVF-embryo transfer at the Department of Reproductive Medicine, Baoding Maternal and Child Health Hospital, between January 2017 and December 2022. Patients were categorized into two groups based on live birth outcome: the live birth group ( n=1 036) and the non-live birth group ( n=756). The dataset was randomly divided into a training set and a validation set in a ratio of 7∶3. Five algorithms were utilized for model development: logistic regression, random forest, extreme gradient boosting (XGBoost), support vector machine, and neural networks. Model performance was assessed using the area under the receiver operating characteristic (AUC) curve, F1 score, and calibration curves. Clinical decision curve analysis (DCA) was employed to evaluate the clinical utility of the models. SHAP values were used to interpret feature importance in the XGBoost model and enhance its explainability. Results:The XGBoost model demonstrated the best performance in predicting CLBR,with accuracy of 72.44%, AUC of 0.775, and F1 score of 0.654, accuracy and F1 score outperforming logistic regression (accuracy was 70.02%, F1 score was 0.585), random forest (accuracy was 71.69%, F1 score was 0.606), support vector machine (accuracy was 70.20%, F1 score was 0.607), and neural network (accuracy was 68.72%, F1 score was 0.560). The calibration curve of XGBoost closely aligned with the diagonal line, indicating that the predicted probabilities were very close to the actual outcomes, demonstrating good calibration. DCA indicated that the XGBoost model provided higher net benefits across a wide range of clinical decision thresholds. SHAP value analysis identified number of previous IVF failures, antral follicle count, anti-Müllerian hormone level, percentage of normal sperm morphology, and sperm DNA fragmentation index as key predictors of CLBR.Conclusion:The XGBoost model exhibits excellent predictive performance and calibration for CLBR, with SHAP values providing important insights into feature importance. This model has the potential to support the development of personalized treatment strategies in clinical practice. However, its generalizability needs to be validated using external datasets to ensure its applicability to diverse populations.

Objective:To retrieve and obtain relevant evidence at home and abroad on the prevention and management of chylous leakage after thyroid cancer surgery, and provide reference for clinical practice.Methods:Based on the 6S pyramid model system, evidence related to the prevention and management of chylous leakage after thyroid cancer surgery in the databases of UpToDate, BMJ best practice, Guidelines International Network (GIN), National Guideline Clearinghouse (NGC), PubMed, EMbase, Wanfang Database, China National Knowledge Infrastructure (CNKI) and other databases, including guidelines, expert consensus, systematic reviews, evidence summary, clinical decisions, and position statements. The retrieval time was from database establishment to June 1st 2024.Results:A total of 16 articles were included, including 5 guidelines, 9 expert consensus, and 2 systematic reviews. The 30 best pieces of evidence were summarized from 6 aspects: preoperative evaluation and preparation, intraoperative prevention, diagnosis, diet and nutrition management, monitoring and treatment, and health education.Conclusions:This study summarized relevant evidence and provides reference for medical staff to formulate intervention measures and management plans for chyle leakage after thyroid cancer surgery.