H.pylori可塑区tfs3a分泌系统virB2基因功能

目的对virB2基因编码蛋白进行分析,为virB2基因及其编码蛋白功能提供实验依据。方法利用多种生物学软件以及网站对VirB2蛋白的结构和功能进行分析预测,VirB2蛋白序列通过基因推导获得并由生物公司合成,然后通过免疫动物实验制备鼠抗VirB2蛋白多克隆抗体,同时设计进行VirB2蛋白细胞毒试验(MTT法)。结果virB2基因编码蛋白属于疏水性蛋白,为鞭毛样结构,有较强的细胞毒作用。结论对VirB2蛋白的结构和功能进行了分析预测,证明VirB2蛋白在H.pylori相关的致病性特别是引起胃黏膜炎症方面起到一定的作用,能够为研究H.pylori致病机制提供帮助。     

Helicobacter pylori induces an increase in inositol phosphates in cultured epithelial cells

Abstract Helicobacter pylori is a bacterial pathogen of humans that infects the gastric mucosa. This infection has been associated with gastritis, peptic ulcers, and gastric carcinomas. Diverse in vitro studies have described efficient adherence of H. pylori to different types of epithelial cells. Because of its varied effects on host cells, we have analysed signal transduction events in H. pyfori -infected epithelial cells. Our results show that H. pylori induces an increase in inositol phosphates in all cultured epithelial cells used, including HeLa, Henle 407, Hep-2, and the human gastric adenocarcinoma cell line AGS. Bacterial growth medium supernatants induce a similar response in the host cell. The increase in inositol phosphates is not related to redistribution of cytoskeletal proteins such as actin or α-actinin nor tyrosine-phosphorylation of host cell proteins. The inositol phosphate increase is also observed in cells infected with low or non-adherent H. pylori mutants or mutants defective in the vacuolating toxin or urease holoenzyme. These results indicate that inositol phosphate release in H. pytori -infected cells is not dependent on bacterial adherence, and that a soluble bacterial factor, but not the vacuolating toxin or urease holoenzyme, mediates such an effect.     

Probiotic Lactobacillus fermentum UCO-979C biofilm formation on AGS and Caco-2 cells and Helicobacter pylori inhibition

The ability of the human isolate Lactobacillus fermentum UCO-979C to form biofilm and synthesize exopolysaccharide on abiotic and biotic models is described. These properties were compared with the well-known Lactobacillus casei Shirota to better understand their anti-Helicobacter pylori probiotic activities. The two strains of lactobacilli synthesized exopolysaccharide as detected by the Dubois method and formed biofilm on abiotic and biotic surfaces visualized by crystal violet staining and scanning electron microscopy. Concomitantly, these strains inhibited H. pylori urease activity by up to 80.4% (strain UCO-979C) and 66.8% (strain Shirota) in gastric adenocarcinoma (AGS) cells, but the two species showed equal levels of inhibition (~84%) in colorectal adenocarcinoma (Caco-2) cells. The results suggest that L. fermentum UCO-979C has probiotic potential against H. pylori infections. However, further analyses are needed to explain the increased activity observed against the pathogen in AGS cells as compared to L. casei Shirota.     

Inhibition of the alpha- and beta-carbonic anhydrases from the gastric pathogen Helycobacter pylori with anions

The gastric pathogen Helicobacter pylori encodes two carbonic anhydrases (CAs, EC 4.2.1.1), an α- and a β-class one, hpαCA and hpβCA, crucial for its survival in the acidic environment from the stomach. Sulfonamides, strong inhibitors of these enzymes, block the growth of the pathogen, in vitro and in vivo. Here we report the inhibition of the two H. pylori CAs with inorganic and complex anions and other molecules interacting with zinc proteins. hpαCA was inhibited in the low micromolar range by diethyldithiocarbamate, sulfamide, sulfamic acid, phenylboronic acid, and in the submillimolar one by cyanide, cyanate, hydrogen sulfide, divanadate, tellurate, perruthenate, selenocyanide, trithiocarbonate, iminodisulfonate. hpβCA generally showed a stronger inhibition with most of these anions, with several low micromolar and many submillimolar inhibitors detected. These inhibitors may be used as leads for developing anti-H. pylori agents with a diverse mechanism of action compared to clinically used antibiotics.     

Helicobacter pylori urease activates blood platelets through a lipoxygenase‐mediated pathway

The bacterium Helicobacter pylori causes peptic ulcers and gastric cancer in human beings by mechanisms yet not fully understood. H. pylori produces urease which neutralizes the acidic medium permitting its survival in the stomach. We have previously shown that ureases from jackbean, soybean or Bacillus pasteurii induce blood platelet aggregation independently of their enzyme activity by a pathway requiring platelet secretion, activation of calcium channels and lipoxygenase‐derived eicosanoids. We investigated whether H. pylori urease displays platelet‐activating properties and defined biochemical pathways involved in this phenomenon. For that the effects of purified recombinant H. pylori urease (HPU) added to rabbit platelets were assessed turbidimetrically. ATP secretion and production of lipoxygenase metabolites by activated platelets were measured. Fluorescein‐labelled HPU bound to platelets but not to erythrocytes. HPU induced aggregation of rabbit platelets (ED₅₀ 0.28 μM) accompanied by ATP secretion. No correlation was found between platelet activation and ureolytic activity of HPU. Platelet aggregation was blocked by esculetin (12‐lipoxygenase inhibitor) and enhanced ～3‐fold by indomethacin (cyclooxygenase inhibitor). A metabolite of 12‐lipoxygenase was produced by platelets exposed to HPU. Platelet responses to HPU did not involve platelet‐activating factor, but required activation of verapamil‐inhibitable calcium channels. Our data show that purified H. pylori urease activates blood platelets at submicromolar concentrations. This property seems to be common to ureases regardless of their source (plant or bacteria) or quaternary structure (single, di‐ or tri‐chain proteins). These properties of HPU could play an important role in pathogenesis of gastrointestinal and associated cardiovascular diseases caused by H. pylori.     

Prevalence of Helicobacter pylori infection in Latin America and the Caribbean populations: A systematic review and meta-analysis

ObjectiveTo describe the prevalence of Helicobacter pylori (H. pylori) infection in Latin Americaand the Caribbean (LAC), through systematic review and meta-analysis by age groups and gender.MethodsSystematic review and meta-analysis of the population-based observational epidemiological studies carried out in LAC, focused on the prevalence of H. pylori and published until March, 2018. The databases utilized in the search were MEDLINE, SCIELO andPUBMED. The prevalence described in the meta-analysis and 95% confidence intervals were estimated by the random effects model, and weighted by the size of the study.ResultsThe 22 selected studies were carried out in 14 countries of LAC, and included 24,178 individuals. The studies were conducted between 1987 and 2012, and all were representative of at least one city. The prevalence of H. pylori infection was 57.57% (CI95%:50.43;64.72) for all ages; in children and adolescents the prevalence was 48.36% (CI95%:38.03;58.70) and in adults 69.26%(CI95%:64.54;76.99). No differences were observed regarding sex.ConclusionPrevalence of H. pylori infection in LAC is high for all age groups. These data reinforce the necessity of actions towards the prevention and treatment of H. pylori infection for all age groups. Treating H pylori infection in young ages probably will reduce gastric cancer incidence in the future.     

幽门螺杆菌感染胃组织基因芯片生物信息学分析

为探讨幽门螺杆菌感染胃组织后差异基因变化,深入分析参与疾病发生、发展的分子机制。从GEO(Gene Expression Omnibus)数据库下载幽门螺杆菌感染胃组织基因芯片数据(GSE5081),根据胃粘膜组织是否受损分组,分别比较幽门螺杆菌感染者与阴性对照组,获得差异基因并进行功能分析包括GO分析、信号通路分析,基因相互作用及基因共表达,得到重要核心基因,并通过实时定量PCR方法进行验证。结果表明:得到参与幽门螺杆菌感染后上调的44个主要基因,主要涉及的GO分析及信号通路包括免疫反应、炎症反应、抗原提呈、细胞因子通路、因子受体关联,细胞粘附分子等。研究发现核心基因CXCR4,CCL20,JAK3,TNFAIP2,PLEK,HLA-DMA,PTPRC,CXCL13,BCL2A1,并通过实时定量PCR的方法进行部分验证,CXCR4,CXCL5,CXCL2在幽门螺杆菌感染后的胃黏膜组织表达高于对照组。幽门螺杆菌感染后胃粘膜组织引起免疫反应,炎症反应,抗原提呈,因子受体关联,细胞粘附分子通路的激活。同时发现一些主要的趋化因子相关基因CXCR4,CXCL5,CXCL2,CCL20,CXCL1等,涉及增殖,炎症,免疫,凋亡基因JAK3,TNFAIP2,PLEK,HLA-DMA,PTPRC,BCL2A1等的表达上调,并实时定量PCR验证部分相关基因的表达。这些结果为从分子网络机制层面上认识幽门螺杆菌感染提供分析思路及基础。     

埃索美拉唑为基础的四联疗法对消化性溃疡患者幽门螺杆菌根除率、炎性因子及生活质量的影响

目的:探讨埃索美拉唑为基础的四联疗法对消化性溃疡(PU)患者幽门螺杆菌(Hp)根除率、炎性因子及生活质量的影响。方法:选取2016年3月～2018年11月间我院接收的117例PU患者,根据数表法将患者随机分为对照组(n=58)和研究组(n=59),对照组给予以奥美拉唑为基础的四联疗法治疗,研究组给予以埃索美拉唑为基础的四联疗法治疗,比较两组患者临床疗效、Hp根除率、炎性因子、生活质量及不良反应。结果:研究组治疗后总有效率为74.58%(44/59),高于对照组患者的53.45%(31/58)(P0.05)。两组患者治疗后白介素-6(IL-6)、超敏C反应蛋白(hs-CRP)、肿瘤坏死因子-α(TNF-α)较治疗前降低,且研究组低于对照组(P0.05)。研究组的HP根除率高于对照组(P0.05)。两组患者治疗后临床症状、心理情感、社交及任务完成评分均较治疗前升高,且研究组高于对照组(P0.05)。两组不良反应发生率比较无统计学差异(P0.05)。结论:埃索美拉唑为基础的四联疗法治疗PU,可有效改善炎性因子水平、Hp根除率和生活质量,疗效确切,安全性好,临床应用价值较高。     

Helicobacter pylori recrudescence and its influencing factors

Helicobacter pylori (H pylori) is known as one of the most common infectious pathogens, with high infection and recurrence rates worldwide. The prevalence of H pylori is up to 90% in developing countries, while the annual recurrence rate is much higher than that in developed countries. Recurrence can occur either by recrudescence or reinfection. Compared with reinfection, the time window for recrudescence is generally shorter, followed by the recurrence of H pylori–associated diseases in the short‐term. Many factors are involved in the H pylori reinfection, such as the prevalence of H pylori infection, living conditions and economic development, health conditions and so forth. Previous studies focused less on H pylori recrudescence. Therefore, the influencing factors for H pylori recrudescence needed further exploration. This study reviewed the recrudescence of H pylori infection and its influencing factors.     

幽门螺杆菌感染与细胞自噬的研究进展

幽门螺杆菌(Helicobacter pylori,H. pylori)是一种革兰阴性微需氧病原菌,也是定植于人类胃黏膜上皮中最特异的一种致病菌。它与人消化性溃疡、慢性胃炎、胃癌及胃黏膜相关组织淋巴瘤(MALT)等疾病密切相关。此后又发现H. pylori可能是一种兼性胞内菌,该菌可能通过自噬在胃上皮细胞和巨噬细胞中得以生存、繁殖并引起慢性持续性感染。本文根据近年发表的自噬相关文献,对H. pylori感染不同细胞后自噬对其存活的影响以及H. pylori不同配体引起的自噬调节作一综述。