TRAIL胸外段基因克隆及其表达与纯化

目的；克隆TNF相关的诱导凋亡配体胞膜外段基因（the extracellular region of the human TRAIL cDNA,TPAILex），构建表达载体并在大肠杆菌DH5α中高效表达有生物学活性的TRAIL蛋白胞膜外段。方法：抗CD3McAb刺激正常人外周血淋巴细胞，用RT－PCR、巢式－PCR方法从活化的淋巴细胞中获得人TRAIL基因及其胞膜外段cDNA，并克隆到pGEM-T Easy载体中，DNA测序鉴定，将TRAIL的胞膜外段基因插入表达载体pGEX-2T，构建重组表达质粒pGEX/TRAILex，用IPTG诱导表达TRAIL蛋白的胞膜外段，GST－Agarose 4B层析柱纯化GST－TRAILex融合蛋白，Western-blot鉴定纯化产物。结果：抗CD3 McAb能诱导正常人外周血核细胞TRAIL的高表达，成功地克隆了TRAIL胞膜外段基因，构建了重组表达载体pGEX/TRAILex，IPTG诱导后得到高效表达的TRAIL蛋白胞膜外段，经12％SDS－PAGE分析，可观察到一分子量和理论值相符（约54kD）的诱导表达。Kodak软件分析表达GST－TRAIL胞膜外段融合蛋白的表达量占菌体蛋白总量的28％，进一步分析证实TRAIL蛋白的胞膜外段主要以可溶性的形式表达，用GST－Agarose 4B层析柱纯化超声破菌后的上清，每升细菌培养液可得到9.8mg纯度的95％以上的GST－Tex。Western-blot结果证实54kD的表达带可与鼠抗TRAIL McAb起特异性反应。结论：成功地克隆了人TRAIL基因，并应用基因工程技术在大肠杆菌中高效表达人TRAIL蛋白的胞膜外段，为进一步研究TRAIL的功能及其在肿瘤生物治疗中的应用奠定了基础。     

120急救中心护士应具备的素质

120，是医疗急救的代名词，以院外急救为重点，承担现场救护、运转及途中医疗监护等工作。目前在全国已有几十座城市建立了120。正是120的医护人员用自己的实际行动，让生命不再脆弱，让死神望而却步。在各个城市只要有生命的呼唤，就有120的身影，120与死神赛跑，与时间赛跑，他们用无言的行动诠释了自己职业的高尚和伟大。现在笔者对120急救中心护士应具备的素质作以简单总结如下：[第一段]     

病毒性感染性疾病基础与临床研究的最前沿

病毒既为人类“最早的威胁”,亦是“最后的敌人”。目前主要的病毒性感染性疾病是禽流感、HIV感染和肝炎,已造成巨大的生命与经济损失。随着治疗技术和疫苗预防的进步,已取得显著的成效并有可能扩展达到防癌的目标。一、流感病毒的最新动向1·禽流感全球流行现状谷口清洲2003年     

艾滋病370例

我国AIDS和HIV感染率逐年大幅度上升，为提高我国广大医务工作者对AIDS的认识，更好地防治AIDS，现将在西非尼日尔共和国工作期间收治的370例AIDS患者临床资料总结分析如下。     

低碳水化合食物可降低心血管疾病的危险

新的研究表明,用含蛋白质与不饱和脂肪的食物部分替代碳水化合物,可降低血压,降低血脂水平。对减少心血管疾病危险有益。减少饱和脂肪的摄入对预防心血管疾病的意义已得到证实。对一组进食低饱和脂肪食物的人群,如果它们的饮食结构不相同,亦将会得出不同的结果,这项研究是将它们的饮食结构分成三组。第一组:吃高碳水化合物食物;第二组:吃高蛋白食物(其中一半为植物蛋白);第三组:吃高不饱和脂肪食物(其中一半为单不饱和胞肪)。三组均控制总热量以维持体重,持续观察3年。     

低蛋白饮食对环孢素A肾病大鼠肾间质纤维化的作用

Objective To investigate whether low-protein diet has protective effect on the progression of renal interstitial fibrosis in rats with cyclosporine A (CsA)-induced nephropathy. Methods Eighteen male Sprague-Dawley rats were randomly divided into three groups (6 rats in each group). The rats in control group (C group) received common diet; in model group (M group) low-salt diet; in intervention group (Ⅰ group) low-salt and low-protein diet. After diet adaptation period of one week, the rats in C group received subcutaneous injection of olive oil 1 mg/kg daily for 5 weeks, while M group and Ⅰ group subcutaneous injection of CsA (diluted into 25 g/L with olive oil) 1 ml/kg for 5 weeks. All the rats were sacrificed at the end of the 5th week. The food-intake and body weight were measured daily. The creatinine clearance (Ccr) was examined before rats were sacrificed. The semi-quantitative pathological analysis on kidney sections was performed. The mRNA and protein expression of transforming growth factor-β1 (TGF-βI) and type Ⅰ collagen (Col Ⅰ) in kidney tissue was determined with real time PCR and immunohistochemical staining, respectively. Results The food-intake and body weight of rats in M and I groups were significantly lower than those in C group (P<0.05). Compared with C group, the Ccr levels in M and Ⅰ groups were significantly reduced [(0.65±0.15) ml/min, (0.40+0.13) ml/min vs (1.55±0.29) ml/min, P<0.05], the relative fibrosis areas of kidney interstitium in M and I groups were significantly increased (3.60%±0.46%, 3.26%±0.75% vs 0.44%±0.24%, P<0.05), the mRNA and protein expression of TGF-β1 in M and I group was significantly up-regulated (by 2.6 and 3.1 times in mRNA and by 1.5 and 1.6 times in protein, respectively, P<0.05), and the mRNA and protein expression of Col Ⅰ in M and I groups was also significantly up-regulated (by 3.0 and 3.5 times in mRNA and by 2.3 and 2.1 times in protein, respectively, P<0.05). There were no significant differences between M and I groups in every parameters above-mentioned except the rat body weight and Ccr. Both the body weight and Ccr in Ⅰ group were significantly lower than those in M group (P<0.05). Compared with C group, the urine osmotic pressure in M group and in I group were deceased (for M group, P>0.05; for I group, P<0.05). Compared with C group, the serum cholesterol levels in M and I groups were significantly increased (P<0.05), and the serum phosphorus level in I group was significantly decreased (P<0.05). The levels of serum albumin and serum calcium of all three groups had no statistical differences (P>0.05). Conclusion Low-protein diet has no renoprutective effects on the rat model of cyclosporin A nephropathy, on the contrary, may induce body weight loss.     

罗格列酮对糖尿病大鼠肾脏保护作用机制的探讨

目的 探讨罗格列酮对糖尿病大鼠肾脏的保护作用机制。方法 将大鼠分为正常组、糖尿病组、小剂量罗格列酮组、大剂量罗格列酮组。药物干预4周后，检测各组大鼠的血糖（BS）、三酰甘油（TG）、胆固醇（TC）、肌酐（Scr）水平及尿白蛋白定量（24h）。随后处死大鼠，测定肾组织中丙二醛（MDA）含量、总抗氧化能力（T-AOC）及铜锌超氧化物歧化酶（Cu-ZnSOD）、谷胱甘肽过氧化酶（GSH-Px）、核转录因子-κB（NF-κB）的活性。同时留取。肾脏组织作PAS染色行病理检查。RT-PCR和免疫组化法检测。肾组织中单核细胞化学吸引蛋白质1（MCP-1）mRNA及蛋白质的表达。结果 与糖尿病组相比，大剂量罗格列酮干预组BS、TC、TG差异无统计学意义；而。肾组织MDA含量、MCP-1mRNA及蛋白质表达、NF-κB活性差异有统计学意义（P〈0．05），SOD、GSH-Px、T-AOC活性显著上升；肾小球面积及体积减小。结论大剂量的罗格列酮可显著改善糖尿病大鼠的。肾脏损害，其机制可能与其抗氧化，抑制NF-κB活性，降低MCP-1的含量有关。     

从中药穿山甲受热后活性成分变化看其内服方法

穿山甲功效为活血、下乳、消肿、排脓。目前复方中，穿山甲与其它药物同煎内服。现代中药研究表明：穿山甲所含化学成分主要为蛋白质、氨基酸，其中蛋白质含量为92．6％。受热后，蛋白质易变性。氨基酸易分解，导致二者的生物活性改变，疗效亦显著降低。穿山甲在剥离和炮制时已做了热处理。因此复方中的穿山甲不应再与其它药物同时煎煮。而应粉成细末后冲服。     

血脂康对急性冠状动脉综合征患者血清高敏C-反应蛋白、血脂蛋白的影响

目的观察急性冠状动脉综合征(ACS)患者血清高敏C-反应蛋白(Hs-CRP)、血脂水平变化及血脂康的干预情况。方法69例ACS患者随机分为血脂康组(40例)和常规治疗组(29例),治疗前后分别测定Hs-CRP、总胆固醇、甘油三酯、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C);另30名健康人为对照组。结果与对照组比较,ACS患者Hs-CRP水平明显升高,且与心肌损害程度密切相关。血脂康治疗2周,能明显下调ACS患者的Hs-CRP水平。结论血清Hs-CRP水平与ACS的发生、严重程度密切相关,血脂康的抗炎作用在ACS的早期治疗中有重要意义。     

免疫缺陷性皮肤病

20072226HIV-1Tat蛋白促血管生成作用研究进展(综述)/陆丛笑(中国海洋大学医药学院),李静,耿美玉∥中国皮肤性病学杂志.-2007,21(4).-239~241,254Tat蛋白是HIV-1基因组编码的6个调控蛋白中的一个重要调控蛋白,在艾滋病相关卡波西肉瘤的发生发展中起一定作用。从Tat蛋白结构特点