[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.

Objective: To explore the mechanism of Buzhong Yiqi Decoction in modulating macrophage polarization and intervening in autoimmune thyroiditis (AIT) mice. Methods: Using the random number table method, 48 SPF-grade NOD.H-2h4 mice were assigned to the normal, model, low-dose (4.10 g/kg), medium-dose (8.19 g/kg), high-dose group (16.38 g/kg) of Buzhong Yiqi Decoction, and selenium yeast tablet (0.026 mg/kg) groups, with eight mice in each group. All groups, except the normal group, were free to drink high iodine water (0.05% sodium iodide) to prepare AIT mouse models for 8 consecutive weeks. After the modeling was complete, each treatment group was orally administered with the corresponding medication, while the normal and model groups were orally administered with an equal volume of distilled water once a day for 8 consecutive weeks. High-performance liquid chromatography with an oscillometric refractive detector was used to analyze the content of Astragaloside Ⅳ in Buzhong Yiqi Decoction. Hematoxylin and eosin staining was used to observe the pathological morphology of mouse thyroid tissue. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of serum thyroid peroxidase antibody (TPO-Ab), thyroglobulin antibody (TgAb), interleukin (IL)-6, IL-10, and tumor necrosis factor-α (TNF-α). An immunofluorescence assay was used to detect the positive area percentage of M1 and M2 macrophages in mouse thyroid tissue. Flow cytometry assay was used to detect macrophage polarization in mouse spleen tissue. Real-time fluorescence quantitative PCR was used to detect the mRNA expression of nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3 (NLRP3), nuclear factor kappa B inhibitory protein α (IκBα), and nuclear factor-κB (NF-κB) p65 in mouse spleen tissue. Western blotting was used to detect the expression of the phosphorylated IκBα (p-IκBα), phosphorylated NF-κB p65 (p-NF-κB p65), and NLRP3 protein in mouse spleen tissue. Results: The content of Astragaloside Ⅳ in Buzhong Yiqi Decoction was (7.09±0.06) g/L. Compared to the normal group, significant lymphocyte infiltration was observed in the thyroid tissue of mice in the model group. The levels of serum TPO-Ab, TgAb, IL-6, and TNF-α increased (P<0.05). The positive area percentage of M1 macrophages in thyroid tissue increased (P<0.05). The proportion of M1 macrophages and M1/M2 in spleen tissue increased (P<0.05). The relative expression levels of NF-κB p65 and NLRP3 mRNA in spleen tissue increased (P<0.05). The relative expression of p-IκBα, p-NF-κB p65, and NLRP3 proteins increased (P<0.05). Compared to the model group, the inflammation infiltration degree in the thyroid tissue of mice in each dose group of Buzhong Yiqi Decoction and selenium yeast tablet group was reduced, the serum TPO-Ab, TgAb, IL-6, TNF-α content was decreased, the spleen tissue M1/M2 was reduced, the expression of NF-κB p65 mRNA was reduced, and the relative expression levels of p-IκBα, p-NF-κB p65 protein were reduced (P<0.05). The Buzhong Yiqi Decoction high-dose and selenium yeast tablets groups showed an increase in IL-10 content, an increase in positive area percentage of M2 macrophages in thyroid tissue, an increase in M2 macrophages proportion in spleen tissue, and a decrease in NLRP3 mRNA and protein relative expression levels (P<0.05). Conclusion Buzhong Yiqi Decoction may regulate macrophage polarization by inhibiting the NF-κB/NLRP3 signaling pathway, thus improving the inflammatory damage in mice with AIT.

Objective To observe the value of dual-energy CT(DECT)for diagnosing knee gouty arthritis(GA)complicated with knee osteoarthritis(KOA).Methods Forty-eight cases of GA(GA group),30 cases of KOA(KOA group)and 60 cases of GA complicated with KOA(GA+KOA group)were retrospectively enrolled.Clinical and imaging data were compared among 3 groups and between each 2 groups,and the efficacy of DECT for diagnosing knee GA complicated with KOA was analyzed.Results Significant differences of detection rate and CT value of bone marrow edema(BME)were found among 3 groups and between each 2 groups(all P＜0.05),also of the course of disease,serum uric acid(SUA),monosodium urate(MSU)volume,bone erosion score and CT value between GA group and GA+KOA group(all P＜0.05).The area under the curve(AUC)of bone erosion score and CT value for diagnosing knee GA complicated with KOA was 0.779 and 0.824,respectively,and of the combination of the above 5 indexes was 0.898,higher than that of each parameter alone(all P＜0.05).Conclusion DECT was helpful for diagnosing knee GA complicated with KOA,and further combining with clinical parameters could improve its diagnostic efficacy.

The heart serves as the "energy factory" of the human body and is responsible for maintaining blood circulation and oxygen supplies.Its normal functioning thus relies on the generation of substantial amounts of ATP to support its mechanical activities.Under pathological conditions such as myocardial infarction,coronary artery sclerosis,and pulmonary hypertension,however,an insufficient blood supply leads to a reduced oxygen supply,subsequently activating a series of compensatory protective mechanisms.Hypoxia-inducible factor-1α(HIF-1α),as a nuclear transcription factor stably expressed under hypoxic conditions,has been shown to regulate oxygen transport by promoting angiogenesis and vasodilation,and to optimize oxygen utilization by regulating the balance of glucose and lipid metabolisms,thus participating in the regulation of various cardiac diseases.HIF-1α plays a crucial role in regulating cardiac energy metabolism and oxidative stress.This review systematically summarizes recent research regarding the various mechanisms of action of HIF-1α in reprogramming cardiac energy metabolism,explores its potential clinical applications in cardiovascular diseases,and proposes future research directions and possible treatment strategies.By comprehensively summarizing the mechanism of HIF-1α in ischemic heart disease,this article aims to provide new ideas and therapeutic targets for the prevention and treatment of cardiovascular diseases.

Objective:To explore the teaching effectiveness of BOPPPS (bridge-in, objective, pre-assessment, participatory-learning, post-assessment, and summary) teaching model in the undergraduate education of pediatric surgery, compare the BOPPPS teaching model with the traditional teaching model, and examine the differences in students' understanding and mastery of knowledge, as well as their ability improvement.Methods:A total of 621 undergraduate students majoring in pediatrics at Chongqing Medical University in the 2021-2023 academic years were selected as the experimental group, and 552 undergraduate students majoring in pediatrics at Chongqing Medical University in the 2018-2020 academic years were selected as the control group. The traditional teaching method was used in the pediatric surgery teaching of the control group, while the BOPPPS teaching model was used in the experimental group. The two groups were compared based on theoretical examination scores, routine assessment scores, overall performance, and teaching satisfaction. A statistical analysis was conducted using SPSS 24.0 software. Continuous data conforming to a normal distribution were expressed as ( x?±s). Two independent samples mean t test was used for analysis. Results:The experimental group demonstrated significantly higher theoretical examination scores [(70.21±10.03) vs. (62.58±8.40)], routine assessment scores [(96.38±2.10) vs. (84.11±2.02)], and total scores [(80.68±11.30) vs. (71.19±9.50)] compared to the control group ( P<0.05). The overall satisfaction with teaching was significantly higher in the experimental group than in the control group [(91.60±1.75) vs. (90.57±0.80)] ( P<0.05). Conclusions:The application of BOPPPS teaching model in pediatric surgery teaching can improve undergraduate students' self-learning ability, clinical thinking ability, and team cooperation ability, thereby enhancing teaching quality.

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.

Objective To analyze the application effect of blunt dissection for myotomy of incision in single-port thoracoscopic lung wedge resection.Methods Patients who underwent single-port thoracoscopic lung wedge resection in our hospital from January to June 2024 were selected and divided into the observation group(32 cases,received blunt dissection for myotomy of incision during the surgery)and the control group(35 cases,received electrosurgical knife for myotomy of incision during the surgery)according to random number table method.The anesthesia time,operation time,intraoperative blood loss,the earliest time to get out of bed after the operation,the dosage of dezocine,and the resting and cough visual analogue scale(VAS)scores in each postoperative period of patients in the two groups were compared.Results There was no significant difference in terms of anesthesia time,operation time,intraoperative blood loss,or dosage of dezocine of patients between the two groups(P>0.05).The earliest time to get out of bed after operation of patients in the observation group was shorter than that in the control group,with significant difference(P<0.05).No significant difference was observed in resting VAS scores 6 hours or 12 hours after operation of patients in both two groups(P>0.05),but significant differences were found in resting VAS scores 24 hours and 48 hours after operation between the two groups(P<0.05).No significant difference was found in cough VAS scores 6 hours after operation of patients between the two groups(P>0.05),but significant differences were observed in cough VAS scores 12 hours,24 hours,and 48 hours after operation between the two groups(P<0.05).Conclusion Compared with electrosurgical knife for myotomy of incision,blunt dissection for myotomy of incision in single-port thoracoscopic lung wedge resection can reduce postoperative pain,promote postoperative ambulation for patients,which is beneficial to postoperative recovery.

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.