肝硬化患者幽门螺杆菌感染的差异性

观察肝硬变患者幽门螺杆菌感染的差异性及其对病情的影响.结果发现随着肝功能的减退,ChildA组,ChildB组,ChildC组HP的感染率逐步升高,CagA、VacA阳性率也逐步升高,ChildC与各组比较HP的感染率,CagA、VacA的阳性率均显著增高,差异有显著性(P<0.05),胃排空无差异(P>0.05).其余各组间比较无差异(P>0.05).提示肝硬变患者与对照组比较HP的感染率及其菌株无显著差异,胃的排空与HP的感染无关,肝脏功能状况与HP感染相关性有待进一步研究.     

1426例幽门螺杆菌抗体检测的临床价值探讨

目的:探讨I型和II型幽门螺杆菌(Helicobacter Pylori,简称 HP)与小儿腹痛的关系.方法:对1426例腹痛患儿,用免疫印迹法进行分型检测.结果:检出HP阳性590例,占41.37%;其中I型HP感染342例,占57.97%;II型248例,占42.03%.结论:免疫印迹法分型检测HP对临床诊治具有重要的指导意义.     

安徽省宿州地区人群幽门螺杆菌感染率及CagA和VacA的表达

目的探讨安徽省宿州地区人群幽门螺杆菌(Hp)的感染率及毒力CagA和VacA表达与胃部疾病的相关性。方法回顾性分析宿州市立医院2017年1月至2020年7月体检中心及住院患者共66449例,其中体检中心碳呼气试验检测患者33634例,医院就诊患者碳呼气试验检测患者23116例,同期就诊患者血清学Hp抗体检测患者9647例(因消化系病就诊4336例),分别测定Hp感染率、各年龄段感染率及男女性别差异等情况,并探讨Hp毒力CagA和VacA表达与年龄、性别、胃部疾病相关性。结果宿州市健康体检人群Hp感染阳性率为38.9%,男女差异不显著(P=0.10);医院就诊成年患者中碳呼气试验与血清学Hp抗体分型检测的Hp感染阳性率分别为35.72%和36.40%,医院消化系疾病就诊患者Hp抗体分型阳性率为36.53%,差异无统计学意义。Hp感染率与年龄有明显的相关性,婴幼儿及青少年Hp感染率较低,20岁以后感染阳性率逐渐升高,50~59岁年龄段达到高峰;消化科就诊Hp阳性患者中Hp-Ⅰ型比例较低,约为17.97%,但Hp-Ⅰ型患者中合并慢性萎缩性胃炎、消化道溃疡和消化道肿瘤等胃部疾病比例较高。结论安徽省宿州地区患者Hp感染率低于全国平均值,Hp感染与年龄具有相关性,CagA和VacA阳性表达与年龄、胃部疾病具有一定相关性。     

Intracellular VacA Is a Valuable Marker to Predict Whether Helicobacter pylori Induces Progressive Atrophic Gastritis That Is Associated with the Development of Gastric Cancer

VacA was histochemically stained in biopsy specimen and was intracellularly and mainly located in fundic gland area. It is recognized gastric atrophy was observed in the H. pylori-positive patients with intracellular VacA compared with others. The aim of study is to understand the relationship between intracellular VacA and the progression of gastric atrophy that is associated with gastric cancer. Biopsy specimens and sera were obtained from 364 people in their 50s and 60s without gastric cancer diagnosed at first endoscopy undergoing diagnostic endoscopy, for H. pylori infection, histology, and the histochemical status of intracellular VacA using anti-VacA Ab during the follow-up period (mean, 7.3 years). Three hundred eleven of 364 enrolled patients were H. pylori positive and 53 patients were H. pylori negative at first endoscopy. VacA was intracellularly stained with vacuolation and cell destruction in the fundic gland in 98 of 311 H. pylori-positive patients and not stained in another 213 H. pylori-positive patients plus 53 H. pylori-negative patients at first endoscopy. Gastric atrophy has significantly progressed in the H. pylori-positive patints with intracellular VacA with gastric ulcers compared with the others and six gastric cancers have developed in this group during the follow-up period (mean, 7.3 years). Intracellular VacA is a valuable marker to predict whether Helicobacter pylori induces progressive atrophic gastritis that is associated with the development of gastric cancer.     

幽门螺杆菌VacA N端片段通过线粒体途径诱导GES-1细胞凋亡

目的:本研究初步探讨幽门螺杆菌(H.pylori)空泡毒素单一毒力决定簇对GES-1胃黏膜上皮细胞凋亡的影响,为进一步研究H.pylori的致病机制奠定基础。方法:将本课题组已构建的pDsRed-Monomer-C1/VacA N端真核表达载体转染至GES-1细胞中,Western blot鉴定VacA蛋白在细胞中的表达;电子显微镜和中性红摄取试验检测GES-1细胞的空泡样变;Hoechst33342染色、Annexin V-FITC凋亡检测试剂盒以及电镜观察细胞凋亡情况,同时采用分光光度法检测细胞Caspase-9、Caspase-3的活化程度,Rho123检测细胞跨膜电位的变化,ELISA法检测细胞色素C的释放。结果:重组质粒pDsRed-Monomer-C1/VacA转染GES-1细胞24小时后,电镜下可明显观察到空泡样变及核固缩、染色质边聚等凋亡特征,重组质粒组部分细胞发生空泡样变;Hoechst 33342染色后镜下可见明显的核染色质浓缩,Annexin V-FITC凋亡检测试剂盒检测发现重组质粒组细胞凋亡率明显高于对照组(P<0.05);Caspase-9和Caspase-3活化程度呈时间依赖性增加,分别在12小时和24小时达到峰值;Rho123染色发现线粒体跨膜电位明显降低;重组质粒转染组释放细胞色素C的浓度呈时间依赖性正相关,与空质粒组及对照组相比差异显著(P<0.05)。结论:VacA N端片段可诱导GES-1细胞凋亡及空泡样变,VacA蛋白可激活Caspase-9和Caspase-3,且可能主要通过线粒体途径诱导GES-1细胞凋亡。     

Immunology of Helicobacter pylori: insights into the failure of the immune response and perspectives on vaccine studies

Helicobacter pylori infects the stomach of half of the human population worldwide and causes chronic active gastritis, which can lead to peptic ulcer disease, gastric adenocarcinoma, and mucosa-associated lymphoid tissue lymphoma. The host immune response to the infection is ineffective, because the bacterium persists and the inflammation continues for decades. Bacterial activation of epithelial cells, dendritic cells, monocytes, macrophages, and neutrophils leads to a T helper cell 1 type of adaptive response, but this remains inadequate. The host inflammatory response has a key functional role in disrupting acid homeostasis, which impacts directly on the colonization patterns of H pylori and thus the extent of gastritis. Many potential mechanisms for the failure of the host response have been postulated, and these include apoptosis of epithelial cells and macrophages, inadequate effector functions of macrophages and dendritic cells, VacA inhibition of T-cell function, and suppressive effects of regulatory T cells. Because of the extent of the disease burden, many strategies for prophylactic or therapeutic vaccines have been investigated. The goal of enhancing the host's ability to generate protective immunity has met with some success in animal models, but the efficacy of potential vaccines in humans remains to be demonstrated. Aspects of H pylori immunopathogenesis are reviewed and perspectives on the failure of the host immune response are discussed. Understanding the mechanisms of immune evasion could lead to new opportunities for enhancing eradication and prevention of infection and associated disease.     

幽门螺杆菌空泡毒素活性检测方法的比较

目的　分别采用细胞计数法及中性红摄入法 (NRU)检测幽门螺杆菌 (Hp)空泡毒素活性 ,探讨NRU检测空泡毒素活性的应用价值。方法　采用Hp液体培养上清浓缩液作为粗制VacA毒素 ,经倍比稀释(1∶16 0～ 1∶2 )后与胃癌SGC 790 1细胞共同孵育 2 4h。用细胞计数法及NRU同时评价VacA的空泡毒素活性 ,比较两种方法检测VacA空泡毒素活性的差异。结果　经中性红染色后 ,胃癌细胞吸收大量染料 ,空泡更加明显。NRU对空泡毒素活性的判断结果与细胞计数法基本符合 ,细胞计数法显示 1∶2 0～ 1∶2滴度样本均呈阳性结果 ,NRU于 1∶2 0～ 1∶5滴度亦得到阳性结果 ,1∶2滴度组因细胞大量死亡得到“假阴性”结果 (A550 =0 .0 6±0 .0 4 )。 1∶4 0滴度组细胞计数法判断为阴性 ,但NRU检测阳性 (A550 =0 .12± 0 .0 4 ,P <0 .0 5 )。结论　NRU检测空泡毒素活性具有简便、快速的特点 ,有助于对空泡毒素活性的定量评价 ,敏感性高于细胞计数法 ,可广泛用于空泡毒素活性的体外检测     

幽门螺杆菌感染与胃窦炎症病理变化的关系

目的 明确幽门螺杆菌感染（H.pylori)与胃窦炎症病理变化的关系。方法 通过胃镜活检取得胃窦组织标本，在快速尿素酶试验阳性和甲苯胺蓝染色中等阳性的病人中筛检出128例患者，其中包括慢性浅表性胃炎（CG）患者68例，十二指肠球部溃疡（DU）患者30例，胃溃疡（GU）患者30例，对患者的血清进行免疫印迹试验，检测患者的细胞毒素相关蛋白A（CagA)、空泡毒素相关蛋白A（VacA)抗体的情况。结果 胃窦炎症积分在CG、DU、GU患者胃窦炎症均表现为活动性，但是没有明显差异；CagA在所有病例中具有普遍易感性。在GU中出现8例胃窦萎缩，其中4例表现为CagA和VacA均阳性。在DU中未见胃窦萎缩。结论 H pylori相关性胃病中胃窦炎症多表现为活动性，但是没有明显差异。CagA在所有病例中具有普遍易感性，胃窦萎缩多出现在GU患者中并与CagA和VacA均阳性相关。     

慢性萎缩性胃炎胃粘膜病理改变与细胞毒素相关蛋白及空泡细胞毒素的关系

目的研究慢性萎缩性胃炎时胃粘膜不同的病理变化与幽门螺杆菌(Hp)致病因子细胞毒素相关蛋白(CagA)和空泡细胞毒素(VacA)之间的关系.方法 120例Hp阳性的慢性萎缩性胃炎患者按粘膜炎症和粘膜萎缩的程度以及有无肠化生进行分组,并抽取血清,用Western blot法测定血清中特异性抗体CagA(116KD)和VacA(89KD).结果 (1)慢性萎缩性胃炎粘膜炎症的程度与CagA检出的阳性率显著相关,炎症程度严重患者的CagA抗体阳性率显著高于轻度炎症病例(85.1% vs 53.28%,P<0.005);(2)胃粘膜重度萎缩者VacA抗体阳性率显著高于胃粘膜轻度萎缩者(77.8% vs 38.7%,P<0.005);(3)慢性萎缩性胃炎患者VacA抗体阳性者多有肠化生,其肠化生的发生率显著高于VacA抗体阴性患者(84.1% vs 30.3%,P<0.005).结论 CagA的表达同慢性萎缩性胃炎的严重程度密切相关,高阳性率患者的胃粘膜炎症活动度较重,而VacA的表达则同胃粘膜的萎缩及肠化密切相关,胃粘膜重度萎缩与肠化患者VacA的表达率显著高于胃粘膜轻度萎缩与肠化的患者.     

幽门螺杆菌毒力因子与胃癌前变化的研究

目的　研究胃癌前状态和癌前病变伴幽门螺杆菌 (Hp)感染的患者 ,其Hp毒力因子细胞毒素相关蛋白A(CagA)、空泡细胞毒素VacA的表达。方法　常规胃镜检查和活检取得的胃黏膜标本做组织病理检查 ,根据所得结果 ,慢性萎缩性胃炎、胃息肉、残胃、胃溃疡、完全型 /不完全型、大肠 /小肠化生为癌前状态组 (82例 ) ,胃黏膜上皮异型增生为癌前病变组 (2 5例 ) ,单纯慢性浅表性胃炎伴Hp感染患者为对照组 (3 0例 )。用蛋白印迹法 (WesternBlot)测定Hp的CagA、VacA因子。结果　癌前状态组和癌前病变组中 ,CagA、VacA因子的检出率显著高于对照组 (P <0 .0 5 )。结论　幽门螺杆菌CagA、VacA因子与胃癌前状态和癌前病变密切相关