绿色荧光蛋白与鼠盘状结构域受体2融合基因的构建与表达

目的: 构建以绿色荧光蛋白(GFP)为报告基因的重组表达质粒DDR2/pEGFP-N3 并分析该融合基因在活细胞中的表达.方法: 利用反转录PCR扩增鼠盘状结构域受体2(DDR2)cDNA,并重组于pEGFP-N3载体.脂质体体外瞬时转染培养的COS-7细胞,RT-PCR和Western Blot分析该融合基因的表达.结果: 从NIH3T3细胞中克隆到DDR2基因的cDNA,并成功构建DDR2-EGFP融合表达载体.以该质粒转染细胞48 h后,RT-PCR和Western Blot分析表明该融合基因能够在COS-7细胞中正确表达.结论: DDR2/pEGFP-N3融合基因真核表达载体构建成功.该融合蛋白具有DDR2和EGFP双重特性,为进一步研究DDR2的功能提供了工具.     

人akt1基因真核表达载体的构建及表达

目的:构建带有flag标签的人akt1及豆蔻酰化akt1真核表达载体,并检测其在真核细胞中的表达及对细胞周期的影响.方法:应用RT-PCR的方法从人牙髓组织mRNA中扩增出akt1和myr-akt1基因,并在其C-末端带上含24bp的flag标签,克隆到pMD-18T载体并测序,再亚克隆至真核表达载体pcDNA3.1( ),酶切鉴定正确后采用脂质体法瞬时转染COS-7细胞,Western blot检测flag-akt1及flag-myr-akt1在细胞中的表达,同时流式细胞术观察细胞周期.结果:测序证实从人牙髓组织mRNA中扩增出的flag-akt1及flag-myr-akt1融合基因的序列以及读框全部正确;脂质体法转染COS-7后检测到预期目的蛋白的表达.流式细胞术的结果显示当AKT1过度表达以及过度活化时对COS-7细胞周期未产生明显影响.结论:成功构建了C-末端带flag标签的akt1及豆蔻酰化akt1真核表达载体,并使其在真核细胞COS-7中表达.但是AKT1对细胞周期的调控有待进一步研究.     

中药蜡疗结合康复护理对气滞血瘀型腰椎间盘突出症患者腰椎功能的影响

目的 探讨中药蜡疗结合康复护理对气滞血瘀型腰椎间盘突出症患者腰椎功能的影响。方法 选择本院于2019年2月—2020年3月间收治的腰椎间盘突出症患者90例,按随机数字表法分为2组,各45例。对照组行常规康复护理联合单纯蜡泥外敷治疗,在此基础上,研究组行中药蜡疗干预。比较2组疼痛程度、腰椎功能、生活质量。结果 干预后,研究组视觉模拟评分法(VAS)评分(0.45±0.06)较对照组(0.85±0.10)低,腰椎日本骨科协会(JOA)评分(25.26±2.05)较对照组(20.68±3.09)高,差异有统计学意义(t=23.009、8.285,P<0.001);研究组躯体疼痛、生理职能、活力、总体健康、生理功能、社会功能评分(71.30±8.21、75.24±7.98、74.12±8.65、67.59±9.94、79.30±10.01、85.60±6.85)较对照组(62.20±7.28、57.54±6.20、58.89±7.02、49.01±8.04、72.23±8.32、79.15±5.85)高,差异有统计学意义(t=5.563、11.750、9.171、9.749、3.644、4.803,P<0.05)。结论 中药蜡疗结合康复护理可有效减轻气滞血瘀型腰椎间盘突出症患者疼痛程度,改善腰椎功能,提高生活质量。     

补体C3对儿童脓毒症患者预后的预测价值

目的 分析补体C3在脓毒症患儿中的变化情况及其与脓毒症病情严重程度的关系,探讨补体C3对脓毒症患儿死亡的预测价值。方法 回顾性收集2019年11月—2021年9月湖南省儿童医院重症医学科收治的529例脓毒症患儿为研究对象。按诊断脓毒症后28 d预后情况分为存活组(n=471)和死亡组(n=58);按入院诊断脓毒症24h内补体C3的中位数(0.77g/L)分为C3正常组(n=273)和C3降低组(n=256)。比较各组间的临床及实验室指标差异,分析补体C3对脓毒症患儿死亡的预测价值。结果 死亡组补体C3水平较存活组明显降低(P<0.05)。多因素logistic回归分析显示,儿童快速序贯器官衰竭评分(pediatric Sequential Organ Failure Assessment,p-SOFA)得分高和补体C3低与脓毒症患儿死亡密切相关(P<0.05)。受试者操作特征曲线分析显示,p-SOFA与补体C3的联合预测模型曲线下面积为0.852,高于p-SOFA、补体C3单项指标的预测价值(P<0.05)。结论 补体C3可作为脓毒症患儿病情严重程度及预后的评价指标;...     

Identification of blaKPC-2 on different plasmids of three Morganella morganii isolates

Three Morganella morganii strains resistant to carbapenems were recovered from the surgical intensive care unit (SICU) in our hospital. Carbapenemases and extended-spectrum β-lactamases (ESBLs) were respectively detected by the modified Hodge test and the modified Clinical and Laboratory Standards Institute (CLSI) ESBL confirmatory test in all isolates. Amplification of whole-cell and plasmid DNAs extracted from isolates with primers specific for the bla _KPC produced an amplicon confirmed to be bla _KPC-2 by sequence analysis. Pulsed-field gel electrophoresis (PFGE) typing revealed that three isolates belonged to two closely related types. Plasmids electrophoresis and restriction analysis revealed that the bla _KPC-2 was located on different plasmids. The transfer of carbapenem resistance from the three original isolates to Escherichia coli EC600 was successful by conjugation. An examination of the outer membrane proteins showed a lack of a 38-kDa outer membrane protein (OMP) compared with M. morganii susceptible to carbapenems. The production of KPC-2 and ESBLs, combined with OMP deficiency, resulted in high-level carbapenem resistance in the M. morganii strains. The genetic environment around bla _KPC-2 analysis revealed that this β-lactamase was located on the same mobile genetic elements which could transfer between different plasmids.     

旋毛虫成虫可溶性虫体蛋白对葡聚糖硫酸钠诱导的小鼠急性结肠炎的影响

本文探讨旋毛虫成虫可溶性虫体蛋白(Soluble adult proteins,SAP)对葡聚糖硫酸钠(Dextran sulphate sodium,DSS)诱导的C57 BL/6小鼠结肠炎的影响.将36只小鼠随机均分为3组:对照组(PBS 组)、肠炎组(PBS+ DSS组)、虫体蛋白治疗组(SAP+ DSS组).小鼠连续饮用3％DSS溶液7d诱发急性肠炎,对照组给予双蒸水.诱发肠炎的同时,治疗组每天经腹腔注射25μg虫体蛋白,肠炎组注射PBS.每天观察对照组和实验组小鼠的临床表现,并进行临床疾病活动指数(DAI)评分.第7d将小鼠处死,观察结肠肉眼或大体病变及组织病理改变.同时提取小鼠脾淋巴细胞和肠系膜淋巴结细胞,用酶联斑点免疫试验检测细胞因子水平.结果显示,与肠炎组相比,虫体蛋白治疗组的小鼠肠炎症状明显缓解,结肠病理变化较轻,脾淋巴细胞和肠系膜淋巴结细胞的Th1型细胞因子IFN-γ分泌水平下降,Th2型细胞因子IL-4和调节性细胞因子IL-10分泌水平显著升高.表明旋毛虫可溶性成虫虫体蛋白对DSS诱导的C57BL/6小鼠结肠炎有缓解作用.     

微波辐射对PC12细胞突触素Ⅰ表达的影响及其机制

AIM: To investigate the effect of microwave radiation on expression and phosphorylation of synapsin I and to discover the mechanism by research on the change of expression of BDNF and its receptor, TrkB. METHODS: PC12 cells were exposed to microwave with average power density being 30 mW/cm(2). HPLC was used to detect the release of amino acids; RT-PCR, Western blot and immunocytochemistry were used to detect the expressions of synapsin I, BDNF and TrkB; immune co-precipitation was used to study the interaction of BDNF and TrkB. RESULTS: It resulted in the decrease of the release of Asp, Glu, GABA and Gly at 1 h (P<0.01) after radiation. Protein of synapsin I was decreased in 9 h-2 d (P<0.01 or P<0.05); its mRNA was decreased in 3-9 h and increased at 1 d (P<0.01 or P<0.05); its phosphorylation was decreased at 3 h, increased at 1 d, and decreased at 2 d again (P<0.01 or P<0.05) after radiation. Protein of BDNF was decreased at 3 h and increased in 1-2 d (P<0.01 or P<0.05); its mRNA were decreased in 3-9 h, increased at 1d, and decreased at 2 d again (P<0.01 or P<0.05) after radiation. Protein of TrkB was increased in 3 h-1 d (P<0.01 or P<0.05); its mRNA decreased at 3 h and 2 d (P<0.01) after radiation. The interaction between BDNF and TrkB was increased in 3-9 h, but decreased in 1-2 d (P<0.01 or P<0.05) after radiation. CONCLUSION: Microwave radiation can induce the decrease of the release of amino acids and the expression and phosphorylation of synapsin I, and the abnormality of expressions and interaction of BDNF and TrkB in PC12 cells. The factors might play a role in the injury and repair of information transmission in PC12 cells.     

Acupoint electrogymnastics therapy for treatment of apoplectic hemiplegia: a multicenter randomized control study

OBJECTIVE: To observe the therapeutic effect of acupoint electrogymnastics therapy on apoplectic hemiplegia, so as to further popularize this therapy in treatment of apoplectic hemiplegia. METHODS: Using three-central, randomized, controlled, multi-blind methods, 153 cases were divided into an electrogymnastics group (n = 77) and an electroacupuncture group (n = 76). They were treated with acupoint electrogymnastics (AE) and traditional electroacupuncture (TEA), respectively. Shousanli (LI 10) and Waiguan (TE 5) on upper limbs and Yanglingquan (GB 34) and Zusanli (ST 36) on the lower limbs were selected in the two groups. A JD-2008 hemiplegia treatment apparatus was used in the AE group. Fugl-Meyer method was used for assessment of motor function and sense function of the four limbs, balance function and joint activity, etc. before and after treatment. RESULTS: In the motor function and sense function of four limbs, balance function and joint activity in the two groups had significant improvement after treatment (both P < 0. 001) with no significant difference between the two groups (P > 0.05). CONCLUSION: Both acupoint electrogymnastics (AE) and traditional electroacupuncture (TEA) have a similar therapeutic effect on apoplectic hemiplegia, but AE has the advantages of safety, reliability and convenience in operation, so it is suitable for long-term rehabilitation treatment of the patient and for popularization in hospital of basic level.     

经鼻咽旁插植组织间后装治疗在鼻咽癌放疗后程加量中的应用

背景与目的:后装近距离治疗常常用于早期鼻咽癌放射治疗后程加量,但常规施源器置入方法定位不准、重复性差,无法向咽旁追量.本研究采用存鼻窦内窥镜引导下进行咽旁插植后装放疗的新方法,旨在解决常规后装治疗的上述缺陷.方法:选择2005年9月至2006年8月初治和复发鼻咽癌外照射后仍有鼻咽粘膜下肿瘤残留超过1 cm或/和咽旁浸润但侵犯范围较局限的患者23例,在鼻窦内窥镜引导下经鼻腔进行鼻咽及咽旁施源器插植术,术后经CT扫描定位并确认插植位置合格后,利用三维近距离治疗系统进行鼻咽、咽旁肿瘤靶区勾画、剂量优化和组织问近距离治疗,测量治疗前后施源器植入的深度,并观察近距离治疗的疗效及并发症.结果:施源器均准确插入肿瘤区,插植位置100%合格.插人和拔出施源器时粘膜下植入深度分别为(9.59±2.72)mm和(9.43±2.30)mm,两者差异无统计学意义(t=0.23,P＞0.05);漂移长度为(0.75±0.75)mm.治疗后3个月内肿瘤完全消失,无肿瘤局部复发,无远处转移.随访3～15个月(中位随访时间6个月),随访率100%,无大出血、感染等手术并发症,无腭穿孔、鼻咽坏死等严重后装放疗并发症;3例鼻甲粘连,经分解后完全缓解.结论:经鼻窦内窥镜鼻咽、咽旁插植定位准确,固定良好,安全可行;对鼻咽癌放疗后鼻咽、咽旁残留病灶进行组织问近距离治疗近期疗效良好,无严重近期并发症.