弓形虫表面抗原P30、P22与霍乱毒素A2／B复合基因真核表达质粒的构建

目的选择弓形虫 RH 株主要表面抗原 P30、P22的有效基因片段和霍乱毒素 A_2/B 亚基共同构建在同一真核表达载体 pcDNA3.1(-)上,并保证其连接方向和开放读码框的正确。方法用PCR 技术分别从弓形虫 RH 株基因组 DNA 和 pUAB024-CTXA_2/B 质粒中扩增编码 P30、P22基因片段和 CTXA_2/B,定向重组入 pUC18克隆载体,然后酶切释放 P30-P22-CTXA_2/B 复合基因片段,亚克隆入pcDNA3.1(-)真核表达载体,再经含氨苄青霉素的 LB 培养基筛选、酶切及测序鉴定。结果酶切产物经电泳显示条带清晰,P30、P22和 CTXA_2/B 基因片段的泳动位置分别在786bp、492bp、512bp 的位置,与预计结果一致;测序结果表明插入的基因片段方向及序列均正确。结论成功地构建了真核表达重组质粒 pcDNA3.1-P30-P22-CTXA_2/B,保证了三个基因连接方向,序列及开放读码框的正确,为下一步融合蛋白 P30-P22-CTXA_2/B 在哺乳动物细胞中的表达及动物实验奠定了基础。     

Protective Role of Genetic Polymorphism of Heat Shock Protein 70-2 for Gastric Cancer Risk

Background Heat shock protein 70-2 (HSP70-2) has a cytoprotective role in various conditions and also protects the gastric mucosa. Recently, polymorphism of HSP70-2 at position 1267 was suggested to be associated with carcinogenesis. We investigated the association of this polymorphism with the risk of gastric cancer in the present study. Methods We examined 223 patients (159 men and 64 women, mean age 64.8 years) with gastric cancer who underwent gastrointestinal endoscopy at our department. The controls were 200 age-matched patients (140 men and 60 women) without gastric cancer diagnosed by gastrointestinal endoscopy. Genotyping was done by PCR-based restriction fragment length polymorphism analysis, and the PCR products were digested with PstI. The two allelic forms, corresponding to the presence or absence of the PstI site, were designated as the P1 allele and P2 allele, respectively. Logistic regression analysis was performed to calculate an odds ratios (ORs) for differences of HSP70-2 polymorphism between the two groups. Results Among the 223 patients with gastric cancer, 46 (20.6%) had P1/P1, 177 (79.4%) were P1 carriers, and 6 (2.7%) were P2/P2. In the control group, 33 (16.5%) patients had P1/P1 polymorphism, 167 (83.5%) were P1 carriers, and 12 (6.0%) were P2/P2. The OR for gastric cancer of subjects with P2/P2 polymorphism relative to P1 carriers was 0.43 (95% CI = 0.16–1.17) (P = 0.097). Among females, the OR for gastric cancer of subjects with P2/P2 polymorphism relative to P1 carriers was 0.10 (95% CI = 0.012–0.838) (P = 0.014). This polymorphism was also associated with a lower risk of middle third cancer (OR = 0.13; 95% CI = 0.02–1.00). Conclusions P2/P2 polymorphism of HSP70-2 at position 1267 was associated with a lower risk of gastric cancer in females.     

基质金属蛋白酶9及热休克蛋白60检测对早期胃癌的诊断价值

目的 探讨血浆基质金属蛋白酶9(MMP-9)与热休克蛋白60(HSP-60)联合检测对早期胃癌患者的诊断价值.方法 选取我院收治的临床疑诊早期胃癌136例患者,根据患者确诊诊断分为:对照组(70例,排除胃癌诊断)和研究组(66例,确诊胃癌诊断).检测并比较两组患者MMP-9、HSP-60 、CA19-9、CA72-4以及CEA水平差异.采用Logistic回归分析影响早期胃癌发生的临床因素.运用ROC曲线评估MMP-9与HSP-60对诊断早期胃癌患者的最佳值.结果 研究组血浆MMP-9、HSP-60、CA19-9、CA72-4与CEA水平均高于对照组(P均＜0.05).多因素Logistic回归分析结果显示,血浆MMP-9与HSP-60是早期胃癌发生的危险因素(P＜0.05).血浆MMP-9与HSP-60诊断早期胃癌的ROC曲线下面积分别为0.781和0.667(P=0.024,0.011).当MMP-9≥ 364.5 μg/L和HSP60 ≥2.84 μg/L为cut-off值,对判断早期胃癌发生的灵敏度为53.0％,特异性为81.4％,准确度为67.6％,ROC曲线下面积为0.862.结论 血浆MMP-9与HSP-60联合检测对早期胃癌发生具有较好的诊断价值.     

Pioglitazone, a specific ligand of peroxisome proliferator-activated receptor-gamma, protects pancreas against acute cerulein-induced pancreatitis

ABM: To determine the effect of pioglitazone, a specific peroxisome proliferator-activated receptor-γ(PPARγ) ligand, on the development of acute pancreatitis (AP) and on the expression of heat shock protein 70 (HSP70) in the pancreas. METHODS: AP was induced in rats by subcutaneous infusion of cerulein for 5 h. Pancreatic blood flow was measured by laser Doppler flowmetry. Plasma lipase activity, interleukin-1β (IL-1β) and IL-10 were determined, Pancreatic weight and histology were evaluated and pancreatic DNA synthesis and blood flow as well as pancreatic mRNA for IL-1β and HSP70 were assessed in rats treated with pioglitazone alone or in combination with cerulein. RESULTS: Pioglitazone administered (10-100 mg/kg i.g.) 30 min before cerulein, attenuated dose-dependently the pancreatic tissue damage in cerulein-induced pancreatitis (CIP) as demonstrated by the improvement of pancreatic histology, reduction in plasma lipase activity, plasma concentration of pro-inflammatory IL-1β and its gene expression in the pancreas and attenuation of the pancreatitis-evoked fall in pancreatic blood flow. CIP increased pancreatic HSP70 mRNA and protein expression in the pancreas and this effect was enhanced by pioglitazone treatment. CONCLUSION: Pioglitazone attenuates CIP and the beneficial effect of this pioglitazone is multifactorial probably due to its anti-inflammatory activities, to the suppression of IL-1β and to the over-expression of HSP70. PPARγ ligands could represent a new therapeutic option in the treatment of AP.     

接触性热痛诱发电位在慢性炎性脱髓鞘性多发性周围神经病中的应用

目的:应用接触性热痛诱发电位(CHEPs)技术研究慢性炎性脱髓鞘性多发性周围神经病(CIDP)患者的痛觉传导通路病变特点,探讨CHEPs在CIDP诊断中的应用价值。方法:选取确诊为CIDP的患者21例及对照组32例,应用CHEP刺激器进行刺激,记录Cz/N的潜伏期,分析比较外周神经Aδ纤维及N波峰潜伏期,同时进行神经传导速度的测定并比较其阳性率。结果:(1)对照组CHEPs波形引出率高于患者组。(2)患者组中VAS评分明显低于对照组(P<0.05)。(3)患者组Aδ纤维传导速度明显低于对照组(P<0.05)。(4)患者组上肢Aδ纤维传导速度减慢率较下肢低(P<0.05)。(5)患者组上下肢近端N波峰潜伏期较对照组延长(P>0.05);但患者组上下肢远端N波峰潜伏期较对照组延长(P<0.05)。(6)患者组Aδ纤维CV异常率高于MCV及SCV(P<0.05)。结论:CHEPs能反应CIDP患者的痛觉传导通路的异常部位及损伤程度,且较传统的神经传导速度更敏感,可作为临床诊断CIDP的一项电生理参考指标。     

卡维地洛与美托洛尔治疗高原慢性充血性心力衰竭疗效的对比研究

目的 比较卡维地洛与美托洛尔治疗高原慢性心力衰竭(CHF)的疗效.方法 90例CHF患者随机分成3组:常规治疗组(20例)给予血管紧张素转化酶抑制剂、利尿剂、地高辛等常规心力衰竭治疗.美托洛尔组(34例)、卡维地洛组(36例)在上述治疗基础上分别给予美托洛尔50 mg,2次/d;卡维地洛25 mg,2次/d口服.随访半年,治疗前、后采用超声心动图测定患者心功能并进行疗效观察.结果 治疗后美托洛尔组、卡维地洛组左心室舒张末期内径(LVEDD)[分别为(57.3±6.5)、(57.2±6.9)mm]和左心室收缩末期内径(LVSED)[分别为(46.6±7.0)、(44.0±6.9)mm]显著低于常规治疗组[分别为(64.7±9.1)、(53.4±9.8)mm],左心室射血分数(LVEF)显著高于常规治疗组[分别为(47.5±8.1)%、(52.9±8.5)%、(42.8±9.2)%](P均＜0.05).卡维地洛组LVEF改善优于美托洛尔组(P＜0.05).死亡情况:常规治疗组4例,美托洛尔组1例,卡维地洛组无死亡.美托洛尔组、卡维地洛组病死率均明显低于常规治疗组(P均＜0.05).结论 美托洛尔、卡维地洛均可明显改善高原CHF患者心功能.卡维地洛疗效及耐受性略优于美托洛尔.

Abstract：

Objective To compare the effect of carvedilol and motoprolol on high altitude chronic congestive heart failure (CHF). Methods Ninety patients with high altitude chronic CHF were divided into three groups randomly:Twenty patients in the regular treatment group treated with angiotensin-converting enzyme inhibitor (ACEI) ,diuretics and digoxin; motoprolol (50 mg twice daily) was given in the motoprolol group( 34cases) additional to regular treatment; carvedilol (25 mg twice daily) was given in the carvedilol group(36cases ) additional to regular treatment. All the patients were followed up for six months and measured the changes of cardiac function by echocardiography. Results Left ventricular end-diastolic dimension (LVEDD) was ( 57. 3 ± 6. 5 ) mm and (57.2 ± 6. 9) mm in the carvedilol group and the motoprolol group respectively, and left ventricular end-systolic dimension (LVESD) was (46. 6 ± 7.0) mm and (44. 0 ± 6. 9 ) mm in the carvedilol group and the motoprolol group respectively, which were all significantly smaller than that in the regular treatment group ([64.7 ±9. 1]mm and [53.4 ±9.8]mm for LVEDD and LVESD,respectively) (Ps ＜0.05). Left ventricular ejection fraction (LVEF) in the carvedilol group and the motoprolol group ( [47.5 ± 8. 1] % and [52. 9 ±8.5] % ,respectively) was higher than that in regular treatment group( [42. 8 ±9. 2]% ) (Ps ＜0. 05).The improvement of LVEF in the carvedilol group was better than that in the motoprolol group (P ＜ 0. 05 ). One case died in the motoprolol group and no death in the carvedilol group,4 cases died in the regular treatment group,the mortality in the motoprolol group and the carvedilol group was significantly lower than that in the regular treatment group. Conclusion Carvedilol and motoprolol significantly improved cardiac function in high latitude CHF patients,and the effect of Carvedilol is slightly better than that of motoprolol.     

人工鼻对人工气道患者下呼吸道医院感染扩散拦截作用的研究

目的研究人工鼻对人工气道患者下呼吸道医院感染扩散的拦截作用效果。方法通过使用人工鼻前后医院感染发生率的比较和同一人工鼻不同拦截面的菌株数比较,研究人工鼻对医院感染细菌扩散的拦截作用。结果使用人工鼻前后医院感染发生率和同一人工鼻不同拦截面的菌株数比较,差异均有统计学意义。结论人工鼻对人工气道患者下呼吸道医院感染扩散的拦截作用明显,具有预防医院感染扩散的作用。     

热休克蛋白70含量变化与脓毒症严重程度的关系

目的 探讨脓毒血症患者热休克蛋白(heat shock protein,HSPs)70含量与疾病严重程度相关关系,及其对病情预后的判断价值.方法 选择2009年4月至2010年11月上海市浦东新区公利医院院急诊科收治确诊脓毒症患者120例进行前瞻性研究,按照SAPSⅡ评分法分为轻、中、重3组,每组40例.另选40例年龄、性别、体质量构成与上述各组差异无统计学意义的体检健康人群作为健康对照组.又根据患者最后转归分为死亡组和存活组.患者入院后立即外周静脉采血5 mL,用流式细胞术测定实验组患者外周血中性粒细胞胞内HSPs70的水平,以及血清中降钙素原(PCT)、C-反应蛋白(CRP)含量.单因素方差分析比较各组间HSPs70,PCT、CRP的变化差异.结果 ①脓毒症患者中性粒细胞胞内热休克蛋白(HSPs)70的含量随SAPS Ⅱ评分增高而逐渐升高,轻度、中度、重度脓毒症之间差异具有统计学意义(P<0.05);血清降钙素原(PCT)含量也随SAPSⅡ评分增高逐渐升高,轻度与重度脓毒症之间差异具有统计学意义(P<0.05).②在所有脓毒症患者中,死亡患者均出现在重度组内,与其他标志物相比,死亡患者的热休克蛋白HSPs70的含量比存活患者明显升高(P<0.05).③在脓毒症患者的受试工作者曲线分析中,中性粒细胞胞内热休克蛋白(HSPs)70的含量与PCT、CRP相比,曲线下面积明显增多.结论 热休克蛋白HSPs70对脓毒症患者病情及预后判断有一定的临床价值.

Abstract：

Objective To investigate the relationships between the level of heat shock protein (HSPs) 70 and severity of sepsis in order to find its effects on the status of sepsis and the prognosis of the patients. Methods In total, 120 patients diagnosed to be suffering from sepsis from the emergency department were selected as studying subjects. According to SAPS Ⅱ score, all of these patients were divided into mild, moderate and severe groups (n =40 in each group) , and at the same time, 40 healthy volunteers were studied as controls. The levels of intracellular HSPs70 in peripheral blood neutrophil cells in the mild, moderate and severe groups were determined by using flow cytometry, while serum levels of procalcitonin (PCT) and C-reactive protein ( CRP) were measured as well. Results ① The levels of heat shock proteins HSPs70 in neutrophil cells and PCT in patients with sepsis escalated gradually with the increase in SAPS Ⅱ score, and there were significant different among the mild, moderate, and severe sepsis groups(P<0.05). ② In all septic patients, the death happened only in the severe group. Compared other markers in the survivals , the level of heat shock protein HSPs70 was significantly higher in fatal patients. ③ In curve plot analysis in the patients with sepsis done by the curve of receiver operating characteristic (ROC) , the area of HSPs70 in neutrophil cells under the curve was significantly larger as compared with that of PCT and CRP. Conclusions Heat shock protein HSPs70 has a certain clinical value in determining the severity of disease and prognosis of patients suffering from sepsis.     

热休克蛋白70抗肿瘤免疫的研究进展

热休克蛋白70(HSP70)是热休克蛋白家族中最重要的一族,除了以分子伴侣的形式发挥重要作用外,还在抗原递呈、交叉递呈、激活淋巴细胞、自然杀伤细胞、巨噬细胞以及诱导树突状细胞成熟中发挥重要作用,参与机体特异性免疫和固有免疫,但其在肿瘤免疫中的具体作用机制还有待进一步阐明。