1.Effect evaluation of pharmaceutical service based on root cause analysis combined with KAP theory on decreasing the protocol violations of investigational medicinal products rate in pediatric clinical trials
Chunyan GUO ; Yi ZHANG ; Yuxin YANG ; Yuguang LIANG ; Qian DING ; Qian WANG ; Chengyue ZHANG ; Yu SUN ; Peng GUO
China Pharmacy 2026;37(9):1206-1210
OBJECTIVE To investigate the effect of pharmaceutical services guided by root cause analysis (RCA) in a problem-oriented manner combined with knowledge-attitude-practice (KAP) theory on reducing the incidence of protocol violations of investigational medicinal products in pediatric clinical trials. METHODS A total of 617 participants from 69 drug clinical trial projects conducted in our hospital from January 2016 to December 2020 were selected as the control group, and 868 participants from 72 drug clinical trial projects from January 2022 to December 2025 as the observation group. RCA was performed on the protocol violations of investigational medicinal product in the control group to identify the types and underlying causes. The control group received routine pharmaceutical services for drug clinical trials, while the observation group was provided with precision pharmaceutical services from the three dimensions of knowledge, attitude and practice on the basis of routine pharmaceutical services, according to the root causes identified by RCA. The occurrence of investigational medicinal products protocol violations was compared between the two groups. RESULTS The total incidence of protocol violations of investigational medicinal products, as well as the incidences of minor and major protocol violations, were all significantly lower in the observation group than in the control group ( P <0.001). The main types of protocol violations in both groups included missed/under-/over-dosing of medications, non-adherence to administration time, failure to adjust dosage as required, and combined medication/vaccination in violation of the protocol. Regarding the responsible subjects of protocol violations, the incidences of protocol violations attributed to participants and their guardians as well as investigators and accidental factors were significantly lower in the observation group than in the control group ( P <0.001, P <0.001, P =0.025). However, there were no statistically significant differences in the incidences of protocol violations caused by sponsor-related reasons between the two groups ( P >0.05). CONCLUSIONS Pharmaceutical services led by pharmacists, based on problem-oriented RCA and combined with KAP theory, can effectively reduce the protocol violations of investigational medicinal products rate in pediatric clinical trials, thereby safeguarding the safety and rights of study participants.
2.Epidemic trends and prevention and control effectiveness of notifiable infectious diseases in Yichang City based on interrupted time series
Qian WU ; Hao ZHANG ; Zhongcheng YANG ; Ling ZHOU ; Yi LIANG ; Yajun CAO
Journal of Public Health and Preventive Medicine 2026;37(1):88-92
Objective To analyze the epidemiological characteristics of statutory infectious diseases in Yichang City from 2015 to 2023 and evaluate the effectiveness of non-pharmaceutical interventions (NPIs) in infectious disease prevention and control, and to provide a basis for formulating prevention and control strategies. Methods Descriptive epidemiological methods were used to analyze annual incidence rates. SARIMA and SARIMA intervention models were constructed to predict the incidence rates of infectious diseases. Interrupted time series analysis (ITS) was applied to assess the control effectiveness. Results The average annual incidence rate from 2015 to 2023 was 787.47/100 000, with the top five diseases being influenza, hand-foot-and-mouth disease, hepatitis B, tuberculosis, and diarrheal diseases. The average incidence rate from 2015 to 2019 (654.31/100 000) was significantly higher than that from 2020 to 2022 (489.01/100 000) (χ2= 3 499.6, P < 0.05). The total incidence rate in 2023 (2 396.51/100 000) was significantly higher than the average annual incidence rates from 2015-2019 (χ2= 108 186.1, P < 0.05) and 2020-2022 (χ2= 112 869.4, P < 0.05). SARIMA model results indicated that the actual incidence rate from 2020 to 2022 decreased by 73.49% compared to the predicted rate without intervention, with the highest decline observed in respiratory infectious diseases (79.57%). The SARIMA-intervention model showed a 55.48% relative decrease in the total incidence rate for 2023, with the largest reduction in respiratory infectious diseases (63.28%) and a slight increase in intestinal infectious diseases (5.48%). Conclusion NPIs effectively reduce the incidence of statutory infectious diseases in the short term, especially for acute respiratory and intestinal infectious diseases. However, long-term effectiveness faces challenges, necessitating the development of differentiated prevention and control strategies.
3.Comparison of sleep EEG power spectral density between depressive episode patients and schizophrenia patients with suicidal behavior
Jingwen LIU ; Yunfei ZHOU ; Jingchu HU ; Jiaoyan ZHOU ; Junwei YANG ; Jie LIANG ; Hong XU ; Yu CANG ; Shimeng MA
Sichuan Mental Health 2026;39(1):50-57
BackgroundPatients with depressive episode and schizophrenia have a high risk of suicide. The sleep electroencephalogram power spectral density characteristics of patients with depressive episode accompanied by suicidal behavior and those with schizophrenia may be different, but there is currently a lack of direct comparative studies on these two groups of patients. ObjectiveTo compare the sleep electroencephalogram power spectral density between depressive episode and schizophrenic patients with suicidal behavior, in order to provide references for exploring predictive indicators of suicidal behavior. MethodsFrom June 2018 to December 2020, 20 patients with depressive episode and 20 patients with schizophrenia who had committed suicide within the past month and were treated at the outpatient department of Shenzhen Kangning Hospital were selected. All of them met the diagnostic criteria for depressive episode or schizophrenia as defined in the International Classification of Diseases, tenth edition (ICD-10). Using a random sampling method, 20 volunteers with matching gender and age to the patient groups were selected from the Cuiping community in Shenzhen as the control group. The subjective sleep of the patients was evaluated using the Insomnia Severity Index (ISI), the Dysfunctional Belief and Attitude about Sleep (DBAS), the Disturbing Dreams and Nightmare Severity Index (DDNSI), and the Epworth Somnolence Scale (ESS). The objective sleep of the patients was assessed using polysomnography. The sleep electroencephalogram was filtered and the power spectral density of the brain wave was analyzed and processed for all the subjects. The subjective and objective sleep conditions of the two patient groups were compared, and the sleep electroencephalogram power spectral density of the patient groups and the control group were also compared. ResultsA comparison of subjective and objective sleep conditions between patients with depressive episode accompanied by suicidal behavior and patients with schizophrenia accompanied by suicidal behavior showed no statistically significant differences (P>0.05). Comparisons of sleep electroencephalogram power spectral density in the W stage (average power of α wave, total power of δ wave, average power of δ wave, average power of θ wave), N1 stage (average power of β wave, total power of α wave, total power of δ wave), N2 stage (total power of α wave, average power of α wave, total power of δ wave, average power of δ wave), N3 stage (average power of α wave, average power of δ wave), and R stage (total power of α wave, average power of α wave, total power of δ wave, average power of δ wave) between patients with depressive episode accompanied by suicidal behavior, patients with schizophrenia accompanied by suicidal behavior, and the control group showed statistically significant differences (P<0.05 or 0.01). The total power of δ wave in the W stage and the average power of β wave and δ wave in the N1 stage were higher in two patient groups were higher than those of the control group. The total power of α wave and the average power of α wave in the N2 stage were lower than those of the control group, while the average power of δ wave was higher than that of the control group. The average power of α wave in the N3 stage of both patient groups were lower than that of the control group, while the average power of δ wave was higher than that of the control group. The total power and average power of α wave in the R stage were lower than those of the control group, while the total power and average power of δ wave were higher than those of the control group. All the differences were statistically significant. Patients with depressive episode accompanied by suicidal behavior had higher average powers of α wave, δ wave, and θ wave in the W stage compared with the control group, while the total power of α wave in the N1 stage was lower in the former group. All these differences were statistically significant (P<0.05). ConclusionThe depressive episode patients accompanied by suicidal behavior have highly overlapping sleep electroencephalogram abnormal patterns with those of schizophrenia patients, mainly manifested as a general decrease in α wave power (N2, N3, R stage) and a general increase in δ wave power (W, N1, N2, N3, R stage) as well as β wave power in N1 stage. At the same time, patients with depressive episode accompanied by suicidal behavior also show specific changes, including an increase in the average power of α and θ waves during the wakefulness period (W stage), and a decrease in the total power of α wave in N1 stage. [Funded by Guangdong Province High-level Clinical Key Specialty (with supporting funds from Shenzhen City) (number, SZGSP013); Shenzhen Key Medical Discipline (number, SZXK041); Shenzhen Clinical Medicine Research Center Project (number, 20210617155253001)]
4.The effect of body mass index and inferior pulmonary ligament division on the residual lung expansion after right upper lobectomy: A retrospective cohort study in a single center
Guang MU ; Wenhao ZHANG ; Hongchang WANG ; Yan GU ; Chenghao FU ; Wentao XUE ; Shiyuan XIE ; Tong WANG ; Ke WEI ; Yang XIA ; Liang CHEN ; Jun WANG
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2026;33(02):261-266
Objective To analyze the effect of releasing the lower pulmonary ligament on right residual lung expansion after right upper lobe resection under different body mass index (BMI) levels. Methods The clinical data of patients who underwent thoracoscopic right upper lobe resection in the First Affiliated Hospital with Nanjing Medical University from 2021 to 2022 were retrospectively analyzed. Patients were divided into a group A (17 kg/m2<BMI≤23 kg/m2), a group B (23 kg/m2<BMI≤29 kg/m2) and a group C (BMI>29 kg/m2) according to BMI. The presence of residual cavity was judged by chest X-ray at 7-10 days after operation, the degree of compensation change of the right main bronchus angle was measured, and the changes in lung volume were determined by CT three-dimensional reconstruction. Results A total of 157 patients who underwent thoracoscopic right upper lobe resection were included, including 71 males and 86 females, with an average age of (59.7±11.2) years. There were 50 patients in the group A, 75 patients in the group B, and 32 patients in the group C. In the group A, compared with those without releasing the lower pulmonary ligament, patients with releasing had a lower incidence of postoperative residual cavity (P=0.016), greater changes in bronchus angle (P<0.001), and smaller changes in lung volume (P<0.001). In the group B and C, there was no significant effect of releasing the lower pulmonary ligament on postoperative residual cavity, bronchus angle, and lung volume changes (P>0.05). Conclusion For patients with thin and long body shape and low BMI, releasing the lower pulmonary ligament is helpful to promote the expansion of the residual lung after right upper lobe resection and reduce the occurrence of postoperative residual cavity in patients.
5.Experimental study on interferon-stimulated gene myxovirus resistance protein 2-mediated restriction of tumor cell sensitivity to reovirus oncolysis
LIANG Dan1,2 ; YANG Zailing1,2 ; YU Jiani1,2 ; LI Xinlan2,3 ; SHEN Tao1,2 ; SUN Yongshun2,4 ; WEI Yongzhu2,4 ; ZHAO Xing1,2
Chinese Journal of Cancer Biotherapy 2026;33(2):132-139
[摘 要] 目的:探讨干扰素刺激基因家族成员黏病毒抵抗蛋白2(MX2)在调控肿瘤细胞对呼肠孤病毒(Reo)溶瘤敏感性中的作用及其机制。方法:选取4株具有不同耐药特征的人源肿瘤细胞,通过CCK-8法评估其对Reo的溶瘤敏感性;通过转录组测序筛选出差异表达基因MX2,qPCR法及WB法验证MX2在4株人源肿瘤细胞中的表达;使用siRNA敲低溶瘤低敏感的COC1/DDP细胞中的MX2基因。在细胞感染Reo病毒后,通过CCK-8法检测细胞存活率;qPCR法检测细胞中Reo病毒S1基因表达;免疫荧光法检测细胞内Reo病毒蛋白的积累;半数组织培养感染剂量(TCID50)法测定病毒滴度;流式细胞术分别检测细胞内Reo病毒dsRNA、活性氧(ROS)水平以及细胞凋亡率;透射电镜观察细胞内质网形态变化并采用WB法检测内质网应激相关蛋白(JNK、p-JNK、eIF2α、p-eIF2α、CHOP、PERK)的表达。结果:在4株肿瘤细胞中,SKOV3细胞对Reo溶瘤作用高度敏感,而COC1/DDP、HuH-7SRB及SNU-398细胞均为溶瘤低敏感性。转录组测序结果显示,MX2在溶瘤低敏感肿瘤细胞中的表达水平显著高于溶瘤高敏感细胞(P < 0.01);在溶瘤低敏感性的COC1/DDP细胞中,敲低MX2显著促进Reo病毒复制、诱导细胞凋亡增加,并升高细胞内活性氧水平(均P < 0.001)。透射电镜观察显示,敲低MX2的COC1/DDP细胞感染Reo病毒后出现内质网肿胀、扩张及断裂等典型内质网应激超微结构改变。WB结果显示,内质网应激关键标志物eIF2α/p-eIF2α、PERK、CHOP及凋亡相关调节蛋白JNK/p-JNK的表达均显著上调(P < 0.05或P < 0.01)。结论:肿瘤细胞对Reo的溶瘤敏感性与其细胞内MX2表达水平密切相关。敲低MX2可显著增强Reo在细胞内的复制,进而促进ROS积累,触发内质网应激并促进凋亡。病毒复制增加与细胞凋亡激活的双重作用,最终协同增强Reo的溶瘤作用。
6.Relationship of non-suicidal self-injury behavior with serum lipid levels and thyroid function among college students with depression
CHEN Lu, YANG Zhiqiang, CAO Xiaoping, ZHAO Yanxia, LIANG Shaoying, LUO Yi, LI Hongyu
Chinese Journal of School Health 2026;47(3):394-397
Objective:
To explore the relationship between non suicidal self injury (NSSI) behavior and serum lipid levels as well as thyroid function among college students with depression.
Methods:
A total of 169 college students with depression in the psychiatry departments of tertiary hospitals (grade 3A and 3B) in Ningbo from December 2023 to April 2025 were selected. The Adolescent Self injury Scale (ASIS) was used to assess the presence of NSSI, and participants were accordingly divided into a NSSI group ( n =51) and a non NSSI group ( n =118). General demographic data (including gender, age, and family situation) were collected from both groups. Blood tests were performed to measure lipid profiles [triglyceride (TG), total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C)] and thyroid hormones [triiodothyronine (T3), thyroxine (T4), free triiodothyronine (FT3), free thyroxine (FT4), thyroid stimulating hormone (TSH)]. Multivariate Logistic regression was employed to analyze risk factors for NSSI, and receiver operating characteristic (ROC) curve analysis was used to evaluate the predictive value of serum lipid and thyroid hormone levels for NSSI occurrence in college students with depression.
Results:
The levels of TC, LDL-C, and TSH in the NSSI group were (4.02±0.73) mmol/L, (2.32±0.36) mmol/L, and (6.57±1.95) mU/L , which were significantly higher than those in the non NSSI group [(3.41±0.56) mmol/L, (2.00±0.27) mmol/L, and ( 4.48± 1.09) mU/L, respectively] ( t =5.32, 5.60, 7.20, all P <0.05). Logistic regression analysis revealed that college students from single parent/reconstituted families, those who had experienced school bullying, and those with higher levels of TC, LDL-C, and TSH had a significantly increased risk of engaging in NSSI ( OR =5.22, 6.12, 5.90, 83.64, 3.64, all P <0.05). ROC curve analysis demonstrated that the combined detection of TC, LDL-C, and TSH had high diagnostic efficacy for predicting NSSI in college students with depression, with a sensitivity of 86.3% and a specificity of 94.9%.
Conclusions
NSSI behavior in college students with depression is associated with serum lipid levels and thyroid function. These biomarkers may serve as useful reference indicators for assessing the conditions of these patients.
7.Role and mechanism of mitochondrial calcium uniporter in the cytoskeleton of pancreatic ductal epithelial cells in a mouse model of acute pancreatitis
Qiaofeng CHEN ; Qingzi FU ; Huiying YANG ; Junbo HONG ; Liang ZHU ; Zhenzhen YANG ; Guodu TANG ; Shiyu ZHANG
Journal of Clinical Hepatology 2026;42(2):400-408
ObjectiveTo investigate the effect of mitochondrial calcium uniporter (MCU) on the cytoskeleton of pancreatic ductal epithelial cells in a mouse model of acute pancreatitis (AP) induced by caerulein (CAE), to analyze the role of MCU in the development of AP, and to provide a theoretical basis for clinical treatment. MethodsIn the in vivo experiment, wild-type male C57BL6/J mice, aged 4 weeks, were randomly divided into control group and AP group, with 6 mice in each group. The mice in the AP group were given intraperitoneal injection of CAE to establish a model of AP, and those in the control group were given intraperitoneal injection of an equal volume of normal saline. Serum and pancreatic tissue samples were collected after 24 hours of modeling. HE staining was used to observe pancreatic histopathological changes; Western Blot was used to measure the expression levels of MCU, glutathione peroxidase 4 (GPX4), and acyl-CoA synthetase long chain family member 4 (ASCL4); kits were used to measure the serum level of amylase. In the in vitro experiment, the human pancreatic ductal epithelial cell line HPDE6-C7 was co-cultured with CAE for 24 hours to establish an in vitro AP model, and the cells were divided into control group, CAE group, RR (an MCU activity inhibitor) group, CAE+RR group, Fer-1 (an ferroptosis inhibitor) group, CAE+Fer-1 group, Erastin (an ferroptosis inducer) group, and CAE+Erastin group. CCK-8 assay was used to observe the influence of different agents on cell viability; Western Blot was used to measure the expression levels of MCU, GPX4, and ASCL4; immunofluorescence assay was used to measure reactive oxygen species (ROS), actin cytoskeleton, and monolayer permeability; kits were used to measure the concentrations of malondialdehyde (MDA), glutathione (GSH), Fe2+, and total iron. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for comparison between two groups. ResultsIn the in vivo experiment, compared with the control group, the AP group had significant increases in pancreatic histopathological score, the serum level of amylase, and the expression levels of MCU and ASCL4, as well as a significant reduction in the expression of GPX4 (all P<0.05). In the in vitro experiment, compared with the control group, the CAE group had significant increases in the expression levels of MCU and ASCL4, a significant reduction in the expression of GPX4, and significant increases in the concentrations of Fe2+, total iron, and MDA, the green fluorescence intensity of ROS, and monolayer permeability, as well as a significant reduction in the concentration of GSH (all P<0.05), with the presence of actin cytoskeleton disruption. Compared with the CAE group, the CAE+RR group had a significant increase in the expression level of GPX4, a significant reduction in the expression level of ASCL4, and significant reductions in the concentrations of Fe2+, total iron, and MDA, the green fluorescence intensity of ROS, and monolayer permeability and a significant increase in the concentration of GSH (all P<0.05), with alleviation of actin cytoskeleton disruption. Compared with the CAE group, the CAE+Fer-1 group had significant reductions in the concentrations of Fe2+, total iron, and MDA, the green fluorescence intensity of ROS, and monolayer permeability and a significant increase in the concentration of GSH (all P<0.05), with alleviation of actin cytoskeleton disruption. Compared with the CAE group, the CAE+Erastin group had significant increases in the concentrations of Fe2+, total iron, and MDA, the green fluorescence intensity of ROS, and monolayer permeability and a significant reduction in the concentration of GSH (all P<0.05), with aggravation of actin cytoskeleton disruption. ConclusionDuring the onset of AP, MCU mediates oxidative stress-induced ferroptosis and leads to the disruption of the pancreatic ductal epithelial barrier, which may be one of the possible pathogeneses of AP.
8.Influence of CYP2C19 gene polymorphism on platelet function and inflammatory cytokines and analysis of factors associated with poor prognosis in elderly patients with ischemic stroke
Hai LIANG ; Hong ZHANG ; Runan XIA ; Huijuan CHEN ; Mengyu JIANG ; Fanqin LI ; Panpan DI ; Miao YANG
China Pharmacy 2026;37(6):782-787
OBJECTIVE To investigate the influence of CYP2C19 gene polymorphism on platelet function and inflammatory cytokines in elderly patients with ischemic stroke, and to analyze potential factors associated with poor prognosis. METHODS A retrospective study was conducted on elderly patients with ischemic stroke admitted to our hospital from June 2024 to June 2025, wh o underwent CYP2C19 genotype testing and received antiplatelet therapy with clopidogrel. The levels of platelet function indicators and inflammatory cytokines before and after treatment were compared among patients with different metabolic phenotypes. Based on the prognosis at 6 months post-treatment, patients were divided into poor prognosis group and good prognosis group. Univariate analysis was performed on general data, metabolic phenotype, the levels of platelet function indicators and inflammatory cytokines. Variables with P <0.05 and the levels of inflammatory cytokines before treatment were included in a multivariate Logistic regression analysis to identify independent risk factors for poor prognosis. Multiple linear regression was used to further analyze the relationship between metabolic phenotypes and inflammatory cytokines. RESULTS A total of 448 elderly patients with ischemic stroke were included; among them, 162 cases were normal metabolic phenotype, 218 were intermediate metabolic phenotype, and 68 were poor metabolic phenotype. No rapid or ultrarapid metabolic phenotypes were observed. After treatment, platelet aggregation rate, the levels of P-selectin and platelet activated complex-1 (PAC-1), high-sensitivity C-reactive Protein (hs-CRP), interleukin-1β (IL-1β), IL-6 and tumor necrosis factor-α (TNF-α) in the normal metabolic phenotype group, intermediate metabolic phenotype group, and poor metabolic phenotype group (except for platelet aggregation rate, and the levels of P-selectin and PAC-1 in the poor metabolic phenotype group) were significantly lower than those before treatment in the same group. Moreover, the above indicators in the normal metabolic phenotype group were significantly lower than those in the intermediate and poor metabolic phenotype groups at the corresponding time, and the levels of platelet function indicators in the intermediate metabolic phenotype group were significantly lower than those in the poor metabol ic phenotype group at the corresponding time ( P <0.05). Univariate and multivariate Logistic regression analyses showed that combined with hypertension, combined with diabetes mellitus, and intermediate or poor metabolic genotypes were independent risk factors for poor prognosis in elderly patients with ischemic stroke ( P <0.05). Multiple linear regression analysis showed that serum levels of hs-CRP, IL-1β, IL-6 and TNF-α before treatment were significantly higher in patients with intermediate and poor metabolic genotypes compared to those with normal metabolic genotype ( P <0.05), with a greater magnitude of increase in inflammatory cytokines observed in the patients with poor metabolic genotype. CONCLUSIONS The elderly ischemic stroke patients with CYP2C19 intermediate and poor metabolic genotypes have poor inhibition effect on platelet and higher levels of inflammatory cytokines than normal metabolic genotype; CYP2C19 gene polymorphism, and in combination with hypertension and diabetes, can be used as independent predictors of poor prognosis.
9.Ectopic expression of hemoglobin subunits enhances the in vitro cytotoxicity of CAR-T cells against tumor cells under hypoxic conditions
YANG Jianxun1,2 ; ZHENG Rui3 ; LIANG Sixin3 ; PAN Jie4 ; LI Yanlong5 ; ZHAI Chenxi5 ; ZHAO Xiaojuan2 ; WANG Pengju3 ; DONG Hao4 ; YAN Bo2 ; SUN Zhihong1 ; YANG Angang3
Chinese Journal of Cancer Biotherapy 2026;33(3):233-242
[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。
10.IL-22 impairs NK cell function and promotes cisplatin resistance in bladder cancer cells via activating the STAT3 signaling axis
YANG Yunjie1 ; CHEN Yang2 ; LIU Qi3 ; GUAN Lixian2 ; LIANG Gengqi2
Chinese Journal of Cancer Biotherapy 2026;33(3):280-287
[摘 要] 目的:探讨IL-22通过STAT3信号轴损害NK细胞功能并促进膀胱癌细胞耐药的机制。方法:常规培养T24细胞,用梯度递增法构建耐药T24/顺铂(DDP)细胞。将细胞分为对照组(不处理)、DDP组、IL-22组、IL-22 + DDP组、IL-22 + anti-IL-22组、IL-22 + DDP + Stattic(STAT3抑制剂)组。qPCR法检测各组T24细胞中IL-22、cyclin D1、Bcl-2 mRNA的表达,WB法检测各组细胞中BAX、BCL2和p-STAT3的表达,CCK-8法检测各组细胞的增殖活性,流式细胞术检测各组细胞的凋亡,ELISA检测各组细胞上清液中乳酸脱氢酶(LDH)、TNF-α、IFN-γ、颗粒酶B(GzmB)和穿孔素(PRF)蛋白的水平。结果:T24/DDP细胞对DDP敏感性降低(P < 0.05);其耐药相关基因P-糖蛋白(P-gp)、肺耐药蛋白(LRP)、多药耐药相关蛋白1(MRP1)和IL-22及其受体表达水平均明显升高(均P < 0.05),说明T24/DDP细胞构建成功。与对照组比较,DDP组T24细胞的增殖活力明显降低、凋亡率升高、BAX蛋白表达升高、BCL2表达下降(均P < 0.05);与DDP组比较,IL-22 + DDP组T24细胞的增殖活明显升高、凋亡率明显下降、BAX蛋白表达降低、BCL2表达升高(均P < 0.05),表明IL-22通过调节BAX/Bcl-2的表达促进T24细胞对DDP耐药性。与对照组比较,IL-22组T24细胞总细胞和核中p-STAT3表达水平均明显升高(均P < 0.05);与IL-22组比较,IL-22 + anti IL-22组T24细胞中 p-STAT3水平明显降低(P < 0.05),说明IL-22激活T24细胞中STAT3的磷酸化过程,并促进其转核。与对照组比较,DDP组T24与NK92细胞共培养上清液中LDH、TNF-α、IFN-γ、GzmB及PRF蛋白水平均明显升高(均P < 0.05),与DDP组比较,IL-22 + DDP组共培养上清液中上述蛋白水平均明显降低(均P < 0.05);与对照组比较,IL-22组共培养上清液中LDH、TNF-α、IFN-γ、GzmB及PRF蛋白水平明显降低(均P < 0.05);与IL-22组比较,IL-22 + Stattic组共培养上清液中上述蛋白水平均明显升高(均P < 0.05),说明IL-22可降低NK92细胞对T24细胞的毒性,STAT3抑制剂可逆转此作用。与DDP组比较,IL-22 + DDP组T24细胞增殖活力明显升高(P < 0.05);与IL-22 + DDP组比较,IL-22 + DDP + Stattic组T24细胞增殖活力明显降低(P < 0.05);与DDP组比较,IL-22 + DDP组T24细胞的凋亡率明显升高(P < 0.05);与IL-22 + DDP组比较,IL-22 + DDP + Stattic组T24细胞的凋亡率明显升高(P < 0.05),说明IL-22调控STAT3影响T24细胞DDP耐药性及NK细胞免疫功能。结论: IL-22通过激活STAT3信号轴促进T24细胞的DDP耐药性,抑制NK细胞功能。


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