老年冠心病患者细胞毒素相关蛋白阳性幽门螺杆菌感染与炎症因子的关系

目前国内外多数研究认为幽门螺杆菌(Hp)感染与动脉粥样硬化(AS)相关(1),而Pasceri等进一步研究发现冠心病(CHD)患者细胞毒素相关蛋白(CagA)阳性Hp感染率高于健康对照组,而CagA阴性的Hp感染率在两组间无明显差异[2].因此,进一步研究Hp的毒力因子CagA在冠心病发病中的作用尤为重要.本研究采用酶联免疫吸附法(ELISA)检测150例CHD患者的Hp感染状况以及CagA的阳性率,并在此基础上分析CagA阳性的Hp感染者血清高敏C反应蛋白(hs-CRP)、热休克蛋白60(HSP60)及细胞间黏附因子-1(ICAM-1)的含量变化,为深入探讨CHD的发生、发展提供科学依据.     

幽门螺杆菌外膜泡在动脉粥样硬化形成中的作用

<正>幽门螺杆菌感染与冠心病的发生密切相关,幽门螺杆菌分泌的细胞毒素相关基因A(cytotoxin-associated gene A,CagA)蛋白参与动脉粥样硬化的发生。作为革兰阴性杆菌内毒素的重要载体,外膜泡具有侵袭力强、携带毒素稳定性好的特性。幽门螺杆菌释放的外膜泡携带CagA蛋白突破胃黏膜上皮细胞屏障后入血,进一步作用于血管内皮细胞,通过一系列炎性反应导致血管内皮功能紊乱,促进动脉粥样     

幽门螺杆菌NCTC11637、NCTC11639毒素相关基因A的克隆、序列分析及真核表达载体的构建

目的克隆幽门螺杆菌毒素相关基因A(cytotoxin-associated gene A,CagA)全长序列并进行序列分析,构建表达CagA的真核表达载体,研究CagA调控胃泌素基因的表达。方法PCR扩增出CagA基因全长片段后构建克隆载体pMD18-T/CagA7和pMD18-T/CagA9,测序结果登录GenBank,登录号分别为GQ161098(NCTC11637)和GQ161099(NCTC11639),用测序后筛选的PstI和BamHI内切酶将原核载体上的CagA基因酶切后连接到pcDNA3.1/ZEO(-)真核表达载体上,构建真核表达载体pcDNA3.1ZEO(-)/CagA7和pcDNA3.1ZEO(-)/CagA9,并经双酶切和CagA PCR扩增鉴定,最后转染胃癌细胞,检测胃癌细胞中胃泌素基因的表达。结果序列比对结果显示NCTC11637和NCTC11637 CagA的同源性为88.77%,NCTC11637与已收录的NCTC11637(AB015416)同源性为99.22%,序列分析显示两者均有两个Ca-gA酪氨酸磷酸化位点,转染细胞后CagA上调胃泌素基因的表达。结论成功克隆了CagA全长基因,并构建了含CagA全长基因的真核表达载体,在胃癌细胞内上调了胃泌素基因的表达。     

Cag(+)幽门螺杆菌感染对急性冠脉综合征患者冠状动脉病变的影响

目的探讨Cag(+)幽门螺杆菌(Helicobacter pylori,Hp)感染对急性冠脉综合征患者冠脉动脉粥样硬化的影响及可能机制。方法根据呼气试验及抗Cag抗体水平将确诊为急性冠脉综合征患者144例分为3组:H pylori-CagA-组(n=27)、H pylori+CagA-组(n=63)及H pylori+CagA+组(n=54)。采用Gensini评分来评估患者的冠状动脉粥样硬化的程度。检测患者血脂、血尿酸、血纤维蛋白原、血清低密度脂蛋白、血清超敏C反应蛋白(high sensitive C reactive protein,hsCRP)及氧化低密度脂蛋白(oxidized low density lipoprotein,oxLDL)浓度,并进行比较。结果方差分析显示,3组冠状动脉Gensini评分比较,差异有统计学意义(P0.05)。H pylori+CagA+组冠状动脉Gensini评分明显高于H pylori-CagA-组、H pylori+CagA-组,差异有统计学意义[(69.02±42.52)分vs.(44.48±40.72)分,P0.05;(69.02±42.52)分vs.(44.91±37.65)分,P0.05]。H pylori-CagA-组与H pylori+CagA-组冠状动脉Gensini评分比较,差异无统计学意义[(44.48±40.72)分vs.(44.91±37.65)分,P0.05]。方差分析显示3组血清总胆固醇、低密度胆固醇、载脂蛋白B、oxLDL及hsCRP浓度比较,差异均有统计学意义(P0.05);与H pylori-CagA-组比较,在H pylori+CagA-组及H pylori+CagA+组上述指标明显升高,其中H pylori+CagA+组水平最高,差异有统计学意义(P0.05)。结论 H pylori+CagA+能增加急性冠脉综合征患者冠脉粥样硬化的风险,其可能通过影响脂质代谢、加速低密度脂蛋白的氧化及激活炎症反应来促进冠状动脉的动脉粥样硬化发展。     

幽门螺杆菌免疫分型在胃十二指肠疾病中的应用价值

目的 探讨幽门螺杆菌(Hp)细胞毒素在胃十二指肠疾病中的作用及HP免疫分型的临床应用价值。方法 采用免疫印迹法检测HP感染的胃十二指肠相关疾病的CagA和VacA抗体，并给所选病例三联HP根除方案治疗后，观察其根除率。结果 ①CagA和VacA的阳性率及CagA和VacA双阳性率正常对照组显著低干DU、GU、及GC组(P＜0．05)，而CagA和VacA双阴性率显著高于DU、GU及GC组(P＜0．05)；除DU组CagA阳性率及CagA和VacA的阳性率、CagA和VacA双阳性率显著高于UND组(P＜0．05)外，其余UND、DU、GU及GC组间CagA和VacA双阳性率及双阴性率均无显著性差异(P＞0．05)。②PU较UND的HP根除率高；HP根除成功的病例中CagA阳性率显著高于CagA阴性(P＜0．01)。结论 CagA和VacA与胃十二指肠疾病发生有密切关系，但不能作为判断HP导致特异性胃十二指肠疾病的单一指标。HP免疫分型可能无助干症状及疾病的诊断。同样的根除治疗方案，CagA阳性菌的HP根除率显著高于CagA阴性菌，CagA抗体可作为预测抗菌治疗疗效的有用的指标。     

幽门螺杆菌CagA与宿主细胞相互作用机制的研究进展

细胞毒素相关蛋白A（CagA）是幽门螺杆菌（Helicobacter pylori,H.pylori）感染导致宿主产生炎性反应的重要效应蛋白。H.pylori感染后通过cag PAI编码的Ⅳ型分泌系统将CagA注入宿主细胞内并发生磷酸化,导致细胞内信号传导等一系列的反应。作为H.pylori的一个重要致病因子,CagA蛋白是目前研究的热点之一。本文主要概述了CagA的特征,以CagA易位进入胃黏膜上皮细胞直至引起细胞内信号传导发生改变为主线,综述了近年来与之相关的研究和发现。     

人胃上皮细胞中幽门螺杆菌CagA相关未知磷酸化蛋白和磷酸化位点分析

背景：细胞毒素相关基因A蛋白（CagA）是Ⅰ型幽门螺杆菌（H.pylori）的主要毒力因子。H.pylori胃癌、胃炎相关株和CagA缺失（△CagA）株的蛋白质组学研究尚未发现CagA相关生物标记蛋白。目的：分析H.pyloriCagA阳性株和△CagA株作用后人胃上皮细胞中差异表达的未知磷酸化蛋白和磷酸化位点,为研究CagA的致病机制提供线索。方法：以金属离子亲和吸附富集技术富集经H.pylori标准株和△CagA株作用4h的人胃腺癌细胞株AGS的磷酸化蛋白,双向电泳（2-DE）分离蛋白,飞行时间质谱（TOF-MS）技术鉴定差异蛋白点。结果：H.pylori标准株作用后,AGS细胞FAM50A蛋白276位丝氨酸发生磷酸化,PQBP1蛋白227～251序列中有磷酸化位点。与H.pylori标准株相比,△CagA株作用于AGS细胞可引起至少13种未知磷酸化蛋白的变化,其质谱图中均出现中性丢失峰,其中2种表达量增加,4种表达量降低,6种消失,1种新发生磷酸化。结论：CagA阳性H.pylori感染可致AGS细胞FAM50A和PQBP1蛋白发生磷酸化。所发现的13种未知磷酸化蛋白为揭示CagA的致病机制提供了线索。     

CagA与幽门螺杆菌致病的相关性研究

本文调查了114例胃镜受检者的 Hp 感染率,应用血清 ELISA 技术检测了 Hp 菌株中 CagA 阳性数,并对该蛋白与慢性胃病的关系进行了初步分析。结果发现54例中 Hp 阳性者,CagA 阳性25例;阳性率46.3%;胃炎、十二指肠溃疡和胃癌组织 CagA 阳性率分别为27.3%,71.4%和85.7%(P<0.01)。CagA 阳性菌株感染者胃粘膜的炎症程度、活动度及萎缩、肠化与淋巴滤泡形成数均高于阴性者。证实了 CagA 在 Hp 致病中的重要作用。     

CagA+Hp感染与消化性溃疡

目的本文旨在探讨十二指肠球部溃疡、胃溃疡等与产细胞毒相关蛋白A幽门螺旋杆菌(CagA+Hp)感染的关系.方法分为三组即无症状对照组、十二指肠球部溃疡及胃溃疡组,Hp感染以快速尿素酶试验(RUT)、14C-呼气试验(14C-URT)和(或)病理特殊染色确定,CagA+Hp感染则进一步通过检测血清CagA IgG加以确定.分别计算各组Hp及CagA+Hp的感染率及Ca-gA+Hp占Hp感染的比例,并作组间比较.结果消化性溃疡时,Hp及CagA+Hp的感染率以及CagA+Hp与Hp感染者之比率均明显高于正常对照组,统计学差异显著(P＜0.01).结论Hp与十二指肠球部溃疡及胃溃疡的发生高度相关,其感染率达90%以上,其中产CagA型Hp又占九成以上,可见CagA+Hp感染与消化性溃疡关系密切,这在消化溃疡的防治中应引起足够的重视.     

胃炎及消化性溃疡患者血清Hp CagA IgG的检测

目的 cagA 基因阳性 Hp 感染与慢性胃炎、消化性溃疡的关系。方法用 ELISA 检测55例慢性胃炎和33例消化性溃疡患者血清中 Hp CagA IgG 水平。结果慢性胃炎患者的 CagA IgG 阳性率为47.3％,其阳性血清的平均 CagA IgG 水平为27.9±12.9U/ml,而消化性溃疡患者之阳性率为72.7％,CagA IgG 水平为41.6±18.0U/ml,两组比较相差显著(p<0.05和 p<0.01)。结论 CagA 阳性 Hp 具有致溃疡作用。     

Haptoglobin polymorphism: a key factor in the proatherogenic role of B cells?

B cells play a role in atherosclerosis. B lymphocytes may reduce the progression of vascular disease. Antibody production against modified auto-antigens is an element in the atheroprotective involvement of B lymphocytes. Paradoxical evidence is emerging from animal studies that suggest a proatherogenic B-cell behaviour independently of autoantibody production. One aspect that has received limited consideration is the role of genetic susceptibility modulated by extracellular matrix proteins. Haptoglobin is a polymorphic glycoprotein that binds to CD22 on B lymphocytes. Hp phenotypes show an important molecular heterogeneity. Hp 2-2 has been linked to an increased susceptibility for atherosclerosis. Haptoglobin and its polymorphism play a role in B-cell migration and function. Hp phenotypes may influence B-T cell dialogue and T cell activation. Haptoglobin is involved in the interplay of lymphocytes, neutrophils, and monocytes. Haptoglobin binds to the CD11b/CD18 receptor and to mast cells. HDL particles can become pro-inflammatory through interactions of Hp-Hb complexes with apolipoprotein A1. Haptoglobin is a chemoattractant to pre-B lymphocytes and monocytes. Beyond the conventional view of haptoglobin as a marker of hemolysis, several findings point towards an immunomodulatory effect of haptoglobin in B-cell mediated progression of atherosclerosis. The balance between proatherogenic and protective immunological properties of the different Hp phenotypes determines if lesions progress or regress. Clinical studies indicate a strong association between the Hp 2-2 phenotype and a more frequent onset of diabetic complications and cardiovascular disease. Findings in animal models (where no haptoglobin polymorphism is present) cannot always be extrapolated to humans.     

幽门螺杆菌与动脉粥样硬化血栓形成性脑卒中相关性研究

目的评价幽门螺杆菌(Hp)感染对动脉粥样硬化血栓形成性脑卒中(AT)患者的危险作用。方法选择69例AT患者为病例组,另选择体检者108例作为对照组.记录Hp感染的潜在危险因素及传统动脉粥样硬化危险因素。采用酶联免疫吸附法测定血清Hp特异性抗体IgG。采用logistic回归模型进行多因素分析。结果病例组中男性、高血压、糖尿病及家族脑卒中所占比例明显高于对照组(P<0.05,P<0.01)。病例组感染率高于对照组,但差异无统计学意义(68.1%vs 63.9%,P=0.56 4);校正Hp感染的潜在危险因素及缺血性脑卒中相关危险因素后,两者差异无统计学意义(P=0.698)。结论 Hp感染与AT无明确相关性。     

Haptoglobin genotypes polymorphism as a risk factor for subclinical atherosclerosis in beta-thalassemia major children; a single center Egyptian study

Abstract

Background/Objectives

Haptoglobin (Hp) is an antioxidant protein. Its genotypic polymorphism had been proposed to influence vascular complications among diabetics, but no data are available about this association among thalassemia patients so far. We have investigated the assumption of an association between Hp genotypes and subclinical atherosclerosis among beta-thalassemia major (TM) children.

Methods

One hundred beta-TM children and 70 matched healthy controls were included. Serum ferritin level and fasting lipid profile were assayed. Haptoglobin genotyping was determined by amplification gel electrophoresis. Carotid intima media thickness (cIMT) was measured using high resolution ultrasound.

Results

The relative distribution of the three Hp genotypes among thalassemia group and the control group were 18 and 14.3% for Hp1-1; 38 and 37.1% for Hp2-1; and 44 and 48.6% for Hp2-2 respectively. There was no significant difference between patients and controls regarding Hp genotypes distribution. Hp2-2 genotype TM children had significantly higher cIMT compared to other genotypes (P < 0.0001). Elevated cIMT was significantly represented in Hp2-2 genotype patients (P < 0.0001) who had higher serum ferritin compared to their counterparts (P < 0.05). Hp2-2 patients were five times more likely to suffer from subclinical atherosclerosis than Hp1-1 and six times than Hp2-1 genotype patients (P = 0.008 and 0.001, respectively); a difference that persisted significant after adjustment for some risk factors compared to Hp2-1 patients (OR 3.96; P = 0.02).

Conclusions

Hp2-2 genotype is a significant predictor for premature atherosclerosis in TM children and confers them an increased risk for iron overload.     

幽门螺杆菌感染对冠心病患者氧化低密度脂蛋白及颈动脉硬化的影响

目的探讨幽门螺杆菌(Hp)感染对冠心病患者氧化低密度脂蛋白(oxLDL)及颈动脉硬化的影响及其可能的发病机制。方法选择2006年12月至2007年12月冠心病住院患者159例,其中男115例,女44例,年龄30～88岁,平均(63±10)岁,近期无感染性疾病及慢性炎症性疾病,有典型的心绞痛病史及心电图的改变,冠状动脉造影均显示至少1支冠状动脉管腔内径狭窄≥50%。采用呼气试验来检测患者是否感染Hp,并根据结果将冠心病患者分为3组:阴性感染组,轻度感染组和重度感染组,使用彩色超声检测患者颈动脉内膜中层厚度(IMT)及颈动脉斑块,同时测定血浆高敏C反应蛋白(hsCRP)、尿酸、纤维蛋白原、oxLDL和血脂水平。结果 3组间胆固醇、低密度脂蛋白胆固醇(LDL-C)、hsCRP及oxLDL水平比较,差异均有统计学意义(均为P0.05),且在重度感染组水平最高;阴性感染组、轻度感染组和重度感染组IMT分别为(0.91±0.21)mm、(1.03±0.40)mm和(1.21±0.51)mm,3组间比较,差异有统计学意义(P0.05)。结论 Hp感染能增加冠心病患者颈动脉粥样硬化的风险,并且重度感染可能会伴随更高的风险,Hp感染可能通过影响脂质代谢、加速LDL氧化和激活炎症反应来促进颈动脉粥样硬化发展。     

上海人幽门螺杆菌vac A基因表型与相关胃肠疾病

目的 探讨用ｖａｃＡ基因表型检测对幽门螺力（Ｈｐ）进行分型,并分析不同基因型与相关胃肠疾病之间的关系,方法 对临床胃黏膜标本中Ｈｐ菌株进行分离培养及其总ＲＮＡ抽提,逆转录聚合酶链反应检测ｖａｃＡ基因表型,结果 ５０株临床分离Ｈｐ菌株ｖａｃＡ基因表型ｓｌ／ｍ２型４６株（２％）ｓｌ／ｍｌ型４株（８％）,未发现ｓ２型,结论 上海人Ｈｐ菌株ｖａｃＡ基因绝大多数表达ｓ１／ｍ２型,ｓｌ／ｍ２基因表型分布于所有     

幽门螺杆菌flaA基因的变异

目的鞭毛是幽门螺杆菌（Ｈｐ）的重要侵袭因子,本研究拟检测Ｈｐ鞭毛素Ａ基因的变异性,进而研究其与Ｈｐ致病性的关系．方法我们对５２例胃粘膜活检标本和１８株Ｈｐ临床分离株进行Ｈｐ特异的１６ＳｒＲＮＡ基因和鞭毛素Ａ基因ＰＣＲ扩增和ＰＣＲＲＦＬＰ及ＰＣＲＲＦＬＰＳＳＣＰ分析．结果对４３例Ｈｐ（＋）的ｆｌａＡ基因ＰＣＲＲＦＬＰ（ＨｉｎｄⅢ）大约可分７个型（变异检出率为１６３％）,而ＰＣＲＲＦＬＰＳＳＣＰ可分３７个型（变异检出率为８６０％）,两者有非常显著性差异．结论ｆｌａＡ基因变异性极大,ＰＣＲＲＦＬＰＳＳＣＰ优于ＰＣＲＲＦＬＰ,是检测其变异的有效方法．     

Haptoglobin polymorphism and peripheral arterial occlusive disease.

Haptoglobin (Hp) 2-2 phenotype is a genetic risk factor in coronary atherosclerosis. In this study, haptoglobin phenotypes were determined in 141 patients with peripheral arterial occlusive disease (PAOD) and compared to a reference population (n = 1000). The relative Hp1 allele frequency was decreased among PAOD patients (0.294 vs. 0.403 for the reference population, P < 0.01) due to an overrepresentation of the Hp 2-2 phenotype (50%, odds ratio 1.82 (95% C.I. 1.28-2.60), P < 0.001). This finding was even more pronounced in non-diabetic and in non-smoking PAOD patients (Hp1 allele frequencies: 0.265 and 0.228, respectively). Serum lipids, inflammatory parameters, and blood pressure levels were comparable among the Hp phenotypes, but serum levels of the antioxidant vitamin C were lower in Hp 2-2 patients than in patients with another phenotype (P < 0.05). In PAOD patients with severe atherosclerotic lesions, maximal walking distance of patients carrying a Hp 2-2 phenotype (225-525 m) exceeded that of other Hp phenotypes (50-242 m) (interquartile ranges) (P < 0.05). The findings demonstrate that, despite an increased risk for developing PAOD, the Hp 2-2 phenotype is associated with a longer maximal walking distance which might be attributed to the earlier reported in vitro angiogenic properties of the Hp 2-2 molecule.     

急性心肌梗死患者幽门螺杆菌感染状况及其与血脂代谢的关系

目的　探讨急性心肌梗死 (AMI)患者幽门螺杆菌 (Hp)感染的发生状况及其与血脂代谢的关系。方法　采用酶联免疫吸附法检测 59例 AMI患者及 30例健康对照者血清抗 Hp抗体及抗 Hp毒素相关基因蛋白 (Hp- Cag A)抗体水平 ,分析其阳性率与患者年龄、性别及血脂水平的关系。结果　患者血清抗 Hp抗体及抗 Hp- Cag A抗体阳性率明显高于健康对照组 (P<0 .0 1 ) ,且随增龄而增加 ,但在 55岁以后趋向稳定。血清胆固醇、甘油三酯、脂蛋白 (a)及低密度脂蛋白包括载脂蛋白 A1和载脂蛋白 B水平明显高于健康对照组 (P<0 .0 5) ,而高密度脂蛋白却低于抗体阴性组。结论　 Hp感染可能会通过影响机体脂质代谢 ,促进动脉粥样硬化的形成进而诱发 AMI。     

定量检测幽门螺杆菌细胞毒素相关蛋白基因A方法的建立

目的建立定量检测幽门螺杆菌（Ｈｐ）细胞毒素相关蛋白基因Ａ（ｃａｇＡ）的方法。方法以含ＨｐｃａｇＡ片段的质粒（ｐＭＣ３）作为外参照品，在微孔板中对聚合酶链反应（ＰＣＲ）扩增ｃａｇＡ的产物进行辣根过氧化物酶标记探针的杂交及酶促显色以定量检测ｃａｇＡ阳性Ｈｐ。结果当ＰＣＲ反应体系中原始模板为１～１０５拷贝，循环次数小于或等于２５时，原始模板以相对固定的指数形式增加，在该范围内适宜定量分析。通过对２０份已知Ｈｐ菌株ｃａｇＡ的检测，符合率达１００％，观察结果的敏感度较电泳法提高了２００倍。定量检测不同胃黏膜组织中ｃａｇＡ阳性Ｈｐ，检出底线为６拷贝／毫克组织。结论本方法特异性强，敏感度高，对研究Ｈｐ与消化道疾病的关系及建立定量检测其他微生物的方法均有实用价值     

Relation of haptoglobin phenotype to early vascular changes in patients with diabetes mellitus

Haptoglobin (Hp) is an antioxidant protein and the major susceptibility gene for atherosclerosis in diabetic patients. The effect of Hp phenotype on arterial compliance and metabolic and inflammatory parameters was investigated. Patients were divided into 3 groups according to Hp phenotype of Hp 2-2, Hp 2-1, and Hp 1-1. Arterial elasticity of large and small arteries was evaluated using the pulse-wave contour analysis method. The large-artery elasticity index (LAEI) was lower in patients with Hp 2-2 compared with Hp 1-1 (8.4 +/- 2.3 vs 12.6 +/- 4.1 ml/mm Hg x 100; p <0.0001). The difference in LAEIs between the Hp 2-1 and Hp 1-1 groups was also significant (9.9 +/- 2.6 vs 12.6 +/- 4.1 ml/mm Hg x 100; p = 0.025). The Hp 2-2 and Hp 2-1 groups did not differ from one another. The small-artery elasticity index (SAEI) was significantly lower in patients with Hp 2-2 compared with Hp 1-1 (2.8 +/- 1.0 vs 4.4 +/- 1.9 ml/mm Hg x 100; p = 0.004). Differences in SAEIs between patients with Hp 2-1 and Hp 1-1, as well as those with Hp 2-1 and Hp 2-2, were not detected. Systemic vascular resistance differed significantly across groups, driven by the difference between patients with Hp 2-2 and Hp 1-1. In conclusion, LAEI and SAEI were significantly lower and systemic vascular resistance was higher in homozygotes for the 2 allele (Hp 2-2) compared with patients with Hp 2-1 or Hp 1-1 phenotypes. Differences in arterial elasticity were detected despite the lack of by-phenotype differences in glycemic control, blood pressure, or presence of cardiovascular risk factors.