[摘 要] 目的：为解决野生型B细胞成熟抗原（BCMA）被γ分泌酶切割导致表达不稳定的问题，构建抵抗γ分泌酶切割的BCMA突变体并构建靶细胞，用于评价BCMA CAR-T细胞的杀伤功能。方法：将野生型BCMA的穿膜域替换为人CD8α穿膜域，构建抵抗γ分泌酶切割的BCMA突变体（BCMA-CD8α TM），构建过表达该突变体的U266（U266BCMA Mut）、K562（K562BCMA Mut）、SKOV3（SKOV3BCMA Mut）和CHO（CHOBCMA Mut）细胞；构建装载NFAT-EGFP报告基因的BCMA CAR Jurkat细胞（BCMA-CAR-Jurkat-Reporter）与U266BCMA Mut细胞共培养，采用FCM检测该细胞中EGFP表达水平以指示NFAT激活水平，荧光素酶法检测BCMA CAR-T细胞对Luciferase标记的K562BCMA Mut细胞的杀伤作用，实时无标记动态细胞分析技术（RTCA）检测BCMA CAR-T细胞对SKOV3BCMA Mut和CHOBCMA Mut细胞的杀伤作用。结果：应用γ分泌酶抑制剂LY411575抑制γ分泌酶活性，显著增强野生型U266细胞表面BCMA表达水平，平均荧光强度上调10倍以上；但撤除抑制剂后BCMA表达水平逐渐降低（P<0.01）；BCMA-CD8α TM突变体可抵抗γ分泌酶的切割作用，在U266细胞表面稳定表达（P>0.05）；U266细胞及过表达BCMA-CD8α TM的U266细胞与BCMA-CAR-Jurkat-Reporter细胞共培养后都可激活Reporter系统、增强EGFP表达，但该效应在BCMA-CD8α TM过表达的U266细胞中更显著（P<0.01）；BCMA-CD8α TM在BCMA表达阴性的K562、SKOV3和CHO 3种靶细胞中成功过表达，且在LY411575处理下该突变体的表达水平仅有小幅度升高；荧光素酶法检测结果显示，不同效靶比下，BCMA CAR-T细胞均可特异、高效杀伤过表达BCMA-CD8α TM的K562细胞；RTCA结果显示，不同效靶比下，BCMA CAR-T细胞均可有效识别、杀伤过表达BCMA-CD8α TM的SKOV3和CHO细胞，但同等效靶比下的Mock-T细胞无此效应。结论：本实验构建的BCMA-CD8α TM突变体能够抵抗γ分泌酶的切割，在多种靶细胞表面稳定表达，为评价BCMA CAR-T细胞体外杀伤的有效性和特异性提供多种检测手段。

[摘 要] 目的：探讨miR-192-5p靶向E盒锌指结合同源框2（ZEB2）对胰腺癌PANC-1细胞增殖、迁移、侵袭和上皮间质转化（EMT）的影响及其作用机制。方法：利用TCGA数据库数据分析miR-192-5p和ZEB2在胰腺癌组织中的表达及两者的相关性。采用qPCR法和WB法分别检测人正常胰腺上皮细胞HPNE和胰腺癌PANC-1细胞中miR-192-5p和ZEB2的表达水平。利用脂质体转染技术转染PANC-1细胞，实验分为miR-192-5p mimic组、Mimic NC组、miR-192-5p inhibitor组、Inhibitor NC组、Mimic NC+pcDNA3.1组、miR-192-5p mimic+pcDNA3.1组、miR-192-5p mimic+pcDNA3.1-ZEB2组。CCK-8法、克隆形成、划痕愈合、Transwell实验分别检测转染PANC-1细胞的增殖、克隆形成、迁移和侵袭能力。qPCR法、WB法、双重免疫荧光实验检测PANC-1细胞中ZEB2、E-cadherin、vimentin的表达水平。通过生物信息学网站预测miR-192-5p的靶基因，并利用双荧光素酶报告基因实验验证miR-192-5p对靶基因的调控作用。结果：胰腺癌组织中ZEB2的表达显著高于正常胰腺组织（P<0.01），且胰腺癌组织中miR-192-5p和ZEB2表达呈负相关（r=-0.419，P<0.01）。与HPNE细胞比较，PANC-1细胞中miR-192-5p低表达、ZEB2蛋白高表达（均P<0.01）。miR-192-5p过表达后，PANC-1细胞增殖、克隆形成、迁移和侵袭能力均显著降低，E-cadherin的mRNA和蛋白表达均上调、vimentin和ZEB2 mRNA和蛋白表达均下调；而抑制miR-192-5p后得到了相反的结果（P<0.05或P<0.01）。miR-192-5p靶向调控ZEB2的表达，过表达ZEB2可部分逆转上调miR-192-5p对PANC-1细胞增殖、迁移、侵袭和EMT进程的抑制作用。结论：miR-192-5p通过靶向ZEB2调控胰腺癌PANC-1细胞的恶性生物学行为。

[Abstract] Objective: To investigate the effect of lncRNA SNHG11 on the proliferation, invasion and migration of non-small cell lung cancer (NSCLC) A549 cells and its possible mechanisms. Methods: qPCR was used to detect the levels of lncRNA SNHG11 and miR-193a-5p in human embryonic lung cells (HEL-1) and lung cancer cells (A549, H1299, and HCC827). A549 cells were transfected with SNHG11 small interfering RNA (si-SNHG11), miR-193a mimic or miR-193a inhibitor. The proliferation of A549 cells was detected by CCK-8 assay, migration and invasion of A549 cells were detected by Wound healing and Transwell assay, the protein expression of Ki67 and Cyclin D1 was determined by Western blot, and the targeting relationship between lncRNA SNHG11 and miR-193a-5p was verified by Dual-luciferase reporter experiment. Results: Compared with HEL-1 cells, the expression level of lncRNA SNHG11 was significantly increased while the expression of miR-193a-5p was decreased in lung cancer A549, H1299 and HCC827 cells (all P<0.05). Silencing lncRNA SNHG11 inhibited the proliferation, migration and invasion of A549 cells and reduced the protein expression of Ki67 and Cyclin D1 (all P<0.05). Over-expression of miR-193a-5p inhibited the proliferation, migration and invasion of A549 cells (all P<0.05). lncRNA SNHG11 could targetedly adsorb miR-193a-5p. miR-139a-5p inhibition could partially reverse the effect of silencing lncRNA SNHG11 on the proliferation, invasion and migration of A549 cells (all P<0.05). Conclusion: lncRNA SNHG11 promotes the proliferation, invasion and migration of NSCLC cells by adsorbing miR-193a-5p.

[Abstract] Objective: To study the role of c-Jun N-terminal kinase (JNK) in vitamin E succinate (VES) activating endoplasmic reticulum stress-induced autophagy in human gastric SGC-7901 cells. Methods: SGC-7901 cells were treated with different doses of VES (5, 10, 15, 20 μg/ml) for 24 h, then, qPCR and WB were used to detect the mRNA and protein expressions of autophagy markers LC3 and Beclin-1; Under the action of reticulum stress inhibitor 4-PBA, the fluorescence intensity and distribution of LC3 were observed under laser confocal microscope, and then qPCR was used to detect the mRNA expression of endoplasmic reticulum stress markers GRP78, GRP94 and autophagy markers LC3, Beclin-1. Under the action of the JNK inhibitor SP600125, WB was used to detect the protein expression changes of p-JNK and autophagy marker proteins LC3 and Beclin-1. Results: Compared with the control group, with the increase of the concentration of VES, the mRNA and protein expressions of Beclin-1 and LC3 showed a gradual increase (all P < 0.05), and LC3-Ⅱ/LC3-Ⅰ ratio also increased significantly (P < 0.01); compared with 20 μg/ml VES group, after pretreatment with endoplasmic reticulum stress inhibitor 4-PBA, mRNA expressions of GRP78, GRP94, LC3 and Beclin-1 decreased (all P < 0.01), and the intensity of LC3 punctate aggregation decreased; after pretreatment with JNK inhibitor SP600125, the protein expression levels of p-JNK, LC3 and Beclin-1 were lower than those of 20 μg/ml VES group (all P < 0.01), these results indicated that inhibition of JNK activity could inhibit the occurrence of autophagy. Conclusion: VES can induce autophagy in SGC-7901cells by activating endoplasmic reticulum stress, and JNK participates in the regulation process of endoplasmic reticulum stress on autophagy.

Adult ; Brain ; Brain Diseases ; Heat Stroke ; Humans ; Magnetic Resonance Imaging ; Male ; Retrospective Studies

［摘要］ 目的：探讨miR-627-3p 在食管鳞状细胞癌（esophageal squamous cell carcinoma，ESCC）组织中的表达及其对ESCC 细胞生物学行为的影响。方法：收集2015 年1 月至2015 年10 月河北医科大学第四医院胸外科手术切除的ESCC组织86 例及对 应的癌旁组织标本20 例。通过qPCR法检测miR-627-3p 在86 例ESCC组织及癌旁组织中的表达，分析其表达与ESCC患者临床 病理学指标及预后的关系；利用Kaplan-Meier Plotter 在线数据库进一步分析数据库中hsa-miR-627 的表达与ESCC患者预后的关 系；通过qPCR法检测miR-627-3p 在4 株ESCC细胞系中的表达，选取表达水平最低的食管癌细胞转染miR-627-3p mimic，选取表 达水平最高的ESCC细胞转染miR-627-3p inhibitor，采用CCK-8 法检测细胞的增殖，采用Transwell 实验检测细胞迁移和侵袭；采 用KEGG分析探讨miR-627-3p 可能介导的信号转导通路，并采用qPCR法验证miR-627-3p 对信号通路中关键基因表达的影响。 结果：miR-627-3p 在ESCC组织中的表达明显低于在癌旁组织中的表达，miR-627-3p 的表达与ESCC患者淋巴结转移和临床分 期相关（均P<0.05）；miR-627-3p 高表达的ESCC 患者的5 年生存率明显高于miR-627-3p 低表达的食管癌患者（P<0.05）。ESCC 细胞系KYSE170 中miR-627-3p 表达最低，KYSE30 中miR-627-3p 表达最高；在KYSE170 细胞中转染miR-627-3p mimic 后，细胞 的增殖能力无明显变化（P>0.05），但细胞的迁移（P<0.05）和侵袭能力（P<0.05）显著降低；在KYSE30 细胞中转染miR-627-3p inhibitor 后，细胞的增殖能力无明显变化（P>0.05），但细胞的迁移（P<0.05）和侵袭能力（P<0.05）显著增高。KEGG分析结果显 示，miR-627-3p 介导了多条与肿瘤相关的信号转导通路。结论：miR-627-3p 在ESCC组织中的表达明显低于在癌旁组织中的表 达，其低表达与ESCC患者的不良预后相关，miR-627-3p 抑制ESCC细胞的迁移和侵袭，可能是通过干扰多条与肿瘤相关的信号 通路发挥生物学功能。

［摘要］ 目的：探讨miR-183 对脑胶质瘤细胞放射敏感性的影响。方法：2020 年10 月至2021 年6 月，收集秦皇岛市第一 医院40 例脑胶质瘤组织标本，对T98G 细胞进行梯度剂量X 射线（0、2、4、6 Gy）照射。采用qPCR 检测miR-183、细胞质多腺 苷酸化元件结合蛋白1（cytoplasmic polyadenylation element-binding protein 1，CPEB1）在脑胶质瘤组织、T98G 细胞和经X 射线 照射的T98G 细胞中的表达量。将miR-183 inhibitor 转染T98G 细胞后下调miR-183 表达，经6 Gy X 射线垂直照射，CCK-8 法、 流式细胞术和WB 法，分别检测T98G 细胞增殖能力、细胞凋亡率及BAX 和Bcl2 蛋白表达量。Targetscan 软件预测和双荧光 素酶报告基因实验检测miR-183 与CPEB1 的靶向关系。下调CPEB1 表达后，经6 Gy X 射线照射，分别用CCK-8 法、流式细胞 术和WB 法检测T98G 细胞增殖能力、细胞凋亡率及BAX 和Bcl2 蛋白表达量。将pcDNA-CPEB1 或CPEB1 siRNA 质粒转染 T98G细胞，分别下调或过表达CPEB1 后，检测miR-183 通过CPEB1 对T98G细胞放射敏感性的影响。结果：脑胶质瘤组织和 细胞中miR-183 呈高表达，CPEB1 mRNA 呈低表达。T98G 细胞中miR-183 的表达量随着X 射线放射剂量增加而降低 （P<0.05），CPEB1 表达量随着X 射线放射剂量增加而升高（P<0.05）。6 Gy X 射线照射T98G 细胞后，下调miR-183 可降低细 胞增殖能力、增加细胞凋亡率，而过表达miR-183 则起到相反作用（P<0.05）。miR-183 靶向CPEB1 mRNA 且负调控CPEB1 表 达。下调CPEB1 表达后，经6 Gy X 射线照射可显著提高T98G 细胞增殖能力（P<0.05）、降低细胞凋亡率（P<0.05），miR-183 可 逆转CPEB1 过表达对细胞T98G 放射敏感性的促进作用（P<0.05）。结论：下调miR-183 的表达能够负调控CPEB1，从而增强 脑胶质瘤细胞的放射敏感性。

<b>Objective:b> To explore the long-term effects of maternal pregnancy bisphenol A (BPA) exposure on emotional and behavioral problems appeared in their preschool children. <b>Methods:b> The study sample was a subset of the China-Anhui Birth Cohort Study (C-ABCS). A unified questionnaire was used to collect basic information on both pregnant women and their children. Free BPA concentration in maternal serum was determined by high-performance liquid chromatographytandem mass spectrometry (HPLC-MS/MS). The parent-report version of the Strengths and Difficulties Questionnaire (SDQ) was used to estimate the emotional and behavioral problems in preschool children. A total of 1 713 pairs of mothers and children were included in this study. Association between BPA exposure during pregnancy and the emotional and behavioral problems in preschool children was evaluated with multinomial logistic regression model. <b>Results:b> Prevalence rates in 1 713 preschool children appeared as: 6.48% of emotional problems, 8.11% for conduct problems, 8.35% for hyperactivity/inattention, 2.86% for peer problems, 11.38% for prosocial behaviors and 7.94% for total difficulties. Subjects were divided according to the degrees of exposure and the results showed as: low exposure group (≤0.120 ng/ml), medium exposure group (0.120OR=1.876, 95% CI: 1.161-3.029), more obvious in boys (OR=2.291, 95%CI: 1.126-4.661). <b>Conclusion:b> Maternal exposure to high level of BPA during pregnancy might increase the detrimental effects of abnormal conducts in their preschool children, more obviously seen in boys.
Benzhydryl Compounds/toxicity* ; Child Behavior Disorders/etiology* ; Child, Preschool ; China/epidemiology* ; Cohort Studies ; Emotions/physiology* ; Environmental Exposure ; Environmental Pollutants ; Female ; Gas Chromatography-Mass Spectrometry ; Humans ; Male ; Maternal Exposure ; Mothers ; Phenols/toxicity* ; Pregnancy ; Prenatal Exposure Delayed Effects ; Prevalence ; Risk Factors ; Surveys and Questionnaires ; Tandem Mass Spectrometry

<b>Objective:b> To understand the situation related to health seeking and diagnosis of imported malaria and to provide practical measures for malaria elimination in Jiangsu province. <b>Methods:b> Data on imported malaria cases in Jiangsu province was retrieved in CISDCP from 2014 to 2016. Relevant information on health seeking behavior, diagnosis and treatment of the disease was gathered. <b>Results:b> A total of 1 068 imported cases were reported in Jiangsu province from 2014 to 2016. Except for one malaria case that was caused by blood transfusion, the rest patients were all recognized as 'imported'. Majority of the cases were migrant laborers working in African countries. The accurate rates on the diagnosis of ovale, vivax and quartan malaria and mixed infection were relatively low, as 79.3% (107/135), 29.5% (18/61), 52.9% (18/34) and 0.0% (0/2) at the primary health care settings, respectively. Rate of seeking health care on the same day of onset was more in 2015 than in 2014 and 2016 (χ(2)=18.6, P=0.001). While only 65.4% (699/1 068) of the patients were diagnosed correctly at the primary health care settings. There appeared no statistical difference in the 3-year-study period (χ(2)=5.4, P=0.246). Capacity on 'correct diagnosis' seemed stronger at the CDC than at the hospital levels (χ(2)=13.2, P=0.000; χ(2)=5.4, P=0.020). Totally, 72.7% (32/44) of the severe falciparum malaria cases did not immediately seek for health care when the symptoms started. <b>Conclusions:b> Migrant workers returning from the high endemic malaria areas seemed to have poor awareness in seeking health care services. Capability on correct diagnosis for malaria at the primary health care settings remained unsatisfactory and staff from these settings needs to receive adequate training.
Adult ; China/epidemiology* ; Female ; Human Migration ; Humans ; Malaria/transmission* ; Male ; Middle Aged ; Plasmodium/isolation & purification* ; Prevalence ; Seasons ; Transients and Migrants ; Travel

<b>Objective:b> To understand the epidemiological and molecular characteristics of typhoid and paratyphoid in China from 2009 to 2013, and provide evidence for the prevention and control of typhoid and paratyphoid, the development and improvement of surveillance strategies. <b>Methods:b> Epidemiological analysis was conducted on the incidence data of typhoid and paratyphoid, and related public health emergencies in China during 2009-2013. Pathogen isolation and culture, serologic test were conducted for the typhoid and paratyphoid cases from 13 national surveillance sites. The isolates were subjected to antimicrobial susceptibility testing. Pulsed-field gel electrophoresis (PFGE) was performed for the molecular typing of these isolates. <b>Results:b> The average incidence of typhoid and paratyphoid in China during this period was 1.03/100 000. The reported case number and incidence decreased with year. The provinces reporting high case numbers were Yunnan, Guizhou, Guangxi, Hunan, Zhejiang, Guangdong and Xinjiang. The incidence of age group 0-4 years was highest. The proportion of farmers and children outside child care settings showed an increasing tendency over time. The annual incidence peak was during July-August. Twenty five outbreaks occurred during 2009-2013. The results of pathogen isolation and culture showed that the positive rate was 3.00% (940/31 322), among the positive isolates, the proportion of Salmonella paratyphi A accounted for higher proportion (68.19%, 641/940) compared with Salmonella typhi (31.60%, 297/940). The drug resistances of Salmonella typhi and Salmonella paratyphi varied, but their resistances to nalidixic acid were highest (50.22% and 85.33%) respectively. A certain amount of Salmonella typhi isolates showed the resistance to the 3rd generation cephalosporins. PFGE analysis showed divergent patterns of Salmonella typhi compared with limited patterns of Salmonella paratyphi A. <b>Conclusion:b> The epidemic level of typhoid and paratyphoid in China was relatively low, but the outbreak occurred occasionally. It is necessary to enhance the laboratory-based surveillance, particularly the capability of etiological diagnosis, outbreak investigation, response and antibiotic resistance monitoring, and conduct risk factor investigation in provinces with high incidences in recent years.
Child ; Child, Preschool ; China/epidemiology* ; Disease Outbreaks ; Drug Resistance, Bacterial/genetics* ; Electrophoresis, Gel, Pulsed-Field ; Epidemics ; Farmers ; Humans ; Incidence ; Infant ; Molecular Typing ; Paratyphoid Fever/microbiology* ; Population Surveillance ; Salmonella paratyphi A/isolation & purification* ; Salmonella typhi/isolation & purification* ; Typhoid Fever/microbiology*