幽门螺杆菌的低温保存方法

目的探索幽门螺杆菌实用性的低温保存方法．方法采用以１００ｇ／Ｌ蔗糖、５００ｍＬ／Ｌ小牛血清作为幽门螺杆菌冷冻保存液，对１３８株幽门螺杆菌在－６２℃条件下进行保存，每６ｍｏ进行一次菌株复活，检查菌株存活情况．结果除一株国内分离株（ＣＡＰＭＮ１０７株）在１８ｍｏ时复活失败外，其他菌株在保存２５ａ后均复活良好，其中对７株菌株的观察已达３ａ，仍能成功复活．结论幽门螺杆菌菌株冷冻保存过程中，若使用１００ｇ／Ｌ蔗糖小牛血清保存液（不含二甲基亚砜），配方简单．同时可免去在－２０℃环境的预冻过程，大大简化了操作过程．菌株保存期一般可达２ａ以上，适合各科研单位常规保存幽门螺杆菌菌株     

幽门螺杆菌感染的治疗

幽门螺杆菌感染的治疗陈洁平徐采朴解放军第三军医大学第一附属医院消化科四川省重庆市６３００３８ＳｕｂｊｅｃｔｈｅａｄｉｎｇｓＨｅｌｉｃｏｂａｃｔｅｒｉｎｆｅｃｔｉｏｎｓ／ｔｈｅｒａｐｙＨｅｌｉｃｏｂａｃｔｅｒｐｙｌｏｒｉ主题词螺杆菌感染／治疗螺杆...     

幽门螺杆菌菌株保存及传递

幽门螺杆菌（He动力皿灯火火山，简称HP）是一种微需氧菌，增殖周期长、存活能力差，给有关科研工作带来很多困难．菌株的保存和传递是一项非常基础的工作，但对各种与菌株相关的科研工作来说，又是十分关键的．近年来，国内外在此领域进行了大量深入细致的工作，基本形成了一套切实可行的菌株保存和传递方案．IHp的春活能力工．互间在胃粘膜活检组织块中的存活能力问的定居部位主要在胃粘液层下层，胃粘膜上皮细胞表面．此处的出已接近中性，局部的粘液环境和微需氧条件很适合HP的生存．当胃粘膜后检组织决取出后，Hp仍可存活较长时间…     

幽门螺杆菌菌株存活能力及常温传递方法

目的观察幽门螺杆菌(Helicobacter pylori,Hp)在常温保存液中的存活能力及影响因素,通过实际菌株传递实验,为我国Hp菌株库建立中各地Hp的收集和供应,提供切实可行的菌株传递方案.方法取新鲜培养3d的Hp 3株(NCTC 11637,NCTC11639和CAPMN62),用Hp保存液(含10g/L蔗糖,500mL/L小牛血清)作成1013cfu/L浓度的菌悬液,分别以满管和半管形式装于1.5mL灭菌小管中,经密封后放于37℃恒温箱中,于12,24,36,48,60,72,96,120,144,和168h,按每种各取出一管,接种双份平皿(100mL/L羊血布氏琼脂)后,于微需氧环境培养3d后观察细菌生长情况.另外,将一满管1013cfu/L浓度的Hp用特快专递(EMS)自北京传送到上海,收到后马上进行接种,并连续观察传递管中菌株在不同时间内的存活情况.结果在37℃保存液中Hp至少可存活72h,保存管中通过加满保存液减少空气残留,能明显延长Hp的存活时间;在气温20℃～34℃条件下,经EMS实际传递的菌株存活良好,且在传递到达后可继续存活至55h.结论使用含10%蔗糖的50%小牛血清细菌传送保护液,在常温下通过EMS进行国内Hp的菌株传递能满足我国各地菌株传递的需要,可被全国Hp菌株库采用.     

幽门螺杆菌的固体培养与液体保存

[目的]探索高效的幽门螺杆菌(Helicobacter pylori,Hp)培养和保存方法。[方法]固体培养基采用空肠弯曲菌琼脂加混合抗生素的马血清,利用固体培养基,在微需氧条件下,37℃培养2~3 d,培养出Hp;菌株采用保存液保存于低温液氮条件下,复苏。[结果]成功培养出Hp,且增菌迅速;菌珠保存可成功复苏。[结论]采用固体培养和保存液低温液氮保存,能够较好培养出细菌,且保存后复苏率高。     

幽门螺杆菌的形态变异及返祖

目的幽门螺杆菌（Ｈｐ）在体外很易发生球形变,目前一般认为是不能培养的退化状态．而对Ｈｐ在环境中生存方式和如何感染大量人群仍不清楚,为此本研究对Ｈｐ的形态变化尤其是球形变的返祖进行了研究．方法本研究通过对１８株Ｈｐ在不同条件下的培养,观察其生存及形态变化,并在加有鸡蛋和小牛血清的空弯培养基上及加有血清有布氏肉汤培养基中是进行培养和返祖,观察其变异规律．结果Ｈｐ可在室温液体中繁殖,１８株中观察到３株球形体Ｈｐ在室温下的返祖,除常见球形变外的Ｈｐ杆状弯曲、长杆状、短杆状、长丝体、巨球体、丝球体、原生质体及细小“Ｓ”和弧形变异．并根据Ｈｐ形态变化的顺序及表型,初步发现Ｈｐ由典型“Ｓ”或多螺旋形态变为杆状弯曲、长杆状、短杆状、然后变为球形、长丝体、巨球体、丝球体、原生质体的规律．部分变异体在一定条件下可返祖．结论Ｈｐ易发生多形变化,有一定规律．可在室温下生长繁殖,球形体可在Ｈｐ鸡蛋培养基上或室温液体培养基中返祖．提示Ｈｐ的球形变在其感染和治疗后的复发中起重要作用     

幽门螺杆菌感染诊断方法的评价

幽门螺杆菌感染诊断方法的评价张玲霞张沥ＳｕｂｊｅｃｔｈｅａｄｉｎｇｓＨｅｌｉｃｏｂａｃｔｅｒｐｙｌｏｒｉ；ｈｅｌｉｃｏｂａｃｔｅｒｉｎｆｅｃｔｉｏｎｓ／ｄｉａｇｎｏｓｉｓ；ｕｒｅａｓｅ／ｄｉａｇｎｏｓｔｉｃｕｓｅ主题词螺杆菌，幽门；螺杆菌感染／诊...     

幽门螺杆菌的毒力研究

幽门螺杆菌（周位icr：nzczer八八加，HP是一种生长于胃粘膜表面与粘膜层之间的微需氧菌．自1983年Mhl．we－，－Wxxad首次从胃粘膜中分离出HP以来，大量研究表明该菌与慢性胃炎、消化性溃疡（PU）密切相关，与胃癌也有一定的关系．但人群中Hp感染率为50％，只有少数Hp感染患者发展为PU或胃炎，造成这种反应的差异有二个原因．一是n毒力的大小，二是人体的防御能力．最近研究了产生细胞毒素的HP菌株，比较和探讨了这方面的资料，为HP致病机制的研究开辟了新的途径．HP要侵及胃粘膜需有定植因子（maintenancefactors）和致病因子（po…     

幽门螺杆菌感染对胃粘膜增殖活性的影响

目的观察幽门螺杆菌（Hp）感染与胃粘膜增殖活性的关系．方法慢性胃炎活检标本68例，用W－S银染法、0．25％复红法和HpDNAPCR技术检测Hp，用免疫组化法观察增殖细胞核抗原（PCNA），C-erb-B2和p53基因的表达，按标准化方案对核仁组成区嗜银蛋白（AgNOR）颗粒计数．结果Hp阳性组（30例）和阴性组（38例）中InyNAⅠ，Ⅱ，Ⅲ，Ⅳ级分别为5，7，4，14例和27，11，0，0例，AgNOR计数分别为3．44±1.20个和1．08±0．08个，差异有显著性（P＜0．05）．Hp阳性组中有4例C-erb-B2在新生腺体和不全肠化柱状细胞中局部阳性，Hp阴性组中未见阳性表达结论Hp感染时胃粘膜增殖活性增强，这可能是胃癌发生的高危因素之一．     

幽门螺杆菌与胃癌的关系

目的为了探索幽门螺杆菌（Hp）与胃癌及其病理类型的关系．方法对住院的经内镜及病理证实的23例胃癌，21例慢性浅表性胃炎及26例十二指肠溃疡患者进行冒窦粘膜Hp检测（采用快速尿素酶试验）及血清Hp-IgG抗体测定（采用ELISA法）．结果Hp在慢性浅表性胃炎、十二指肠溃疡、中晚期胃癌的检出率分别为52．4％，88．5％及78．3％．Hp在十二指肠溃疡及慢性浅表性胃炎的感染率具有明显差异（P＜0．01），而Hp在中晚期胃癌及慢性浅表性胃炎的感染率无差异（P＞0．05）．胃腺癌的Hp检出率高于印戒细胞癌的Hp检出率（P＜0．05）．结论①Hp与中晚期胃癌无明显相关性．②胃腺癌患者的Hp检出率较胃印戒细胞癌者的高．     

Helicobacter pylori

PURPOSE OF REVIEW: Helicobacter pylori is an important human pathogen, responsible for most peptic ulcer disease, gastritis and gastric malignancies. H. pylori has several unique features: it is highly adapted for gastric colonization, yet it produces clinical consequences in a small minority, its genome is known, and it is the only bacterium strongly associated with cancer. H. pylori is therefore of great interest to clinicians and researchers of many, often disparate, disciplines. We highlight recent advances in this fast changing field from many different areas. RECENT FINDINGS: The major contentious clinical issues relate to the synergistic gastrotoxic interactions of H. pylori with non-steroidal anti-inflammatory drugs, and a possible association of H. pylori with atherosclerotic events. Accumulating evidence implicates genetic variation in the inflammatory response to H. pylori in the etiology of the increased risk of gastric cancer after H. pylori infection. Studies of pathogenesis have been aided by increasingly sophisticated murine models. The effects in gastric epithelial cells of two of the major virulence factors (genes within the cag pathogenicity island and the vacuolating cytotoxin, VacA) of H. pylori illustrate the complex network of cellular reactions activated by H. pylori. The metabolism of H. pylori is dependent on the availability of hydrogen. SUMMARY: Basic science research into H. pylori continues to elucidate the mechanisms by which H. pylori infection causes disease. These findings have implications for the design of novel therapies and for improving clinical strategies to identify at-risk individuals. Many are also worthy of consideration for other epithelial-microbial interactions.     

Helicobacter pylori

The diagnostics and treatment of Helicobacter pylori infections have substantially changed in recent years. Instead of a general test-and-treat strategy, differentiated treatment methods are increasingly being used. Practical problems in many cases were that a useful combination was often not employed after the failure of an initial antibiotic treatment. In 2009 new guidelines on the diagnostics and treatment of Helicobacter pylori infections were published. Various expert groups from gastro-enterology, microbiology and rheumatology provided new general frameworks and concrete treatment suggestions for Helicobacter pylori infections of the stomach. The statements are grouped according to ?should“ and ?can“ recommendations and the consensus opinion is divided into various subgroups. The new S3 guidelines specify the therapy indications with respect to first and second line procedures and now give different durations of therapy (7 days for first line, 10 days for second line after treatment failure) as well as concrete algorithms. Before treatment two positive diagnostic procedures are required because the prevalence in Germany is decreasing. In addition to the rapid test and histological investigations, the 13-C breath test and stool tests with excellent sensitivity and specificity are also now available. Probiotics can improve therapy success especially for long-term antibiotic regimes and in the future bismuth could again play an increasingly more important role because antibiotic resistance to metronidazol and clarithromycin is increasing.     

Helicobacter pylori

Helicobacter pylori is associated with various gastroduodenal diseases such as peptic ulcer, functional dyspepsia, MALT lymphoma and distal gastric cancer. Diagnosis of H. pylori can be established by non-invasive (^13Curea breath test, stool antigen test, serology) and invasive (histology, rapid urease test, culture) tests. In adults, culture and susceptibility testing should or must be performed after failing of first-line therapy in case of a control endoscopy and before third-line therapy, respectively. Peptic ulcer and gastric MALT lymphoma represent obligatory indications for eradication therapy. Other potential indications are functional dyspepsia, prevention of gastric cancer in individuals being at risk, and before starting treatment with traditional non-steroid antiphlogistics. First-line therapy is performed with a 7-days combination of proton pump inhibitor with clarithromycin and amoxicillin or metronidazole. In second-line therapy levofloxacin and rifabutin are good rescue antibiotics.     

Diagnosis of Helicobacter pylori infection and diseases associated with Helicobacter pylori by Helicobacter pylori outer membrane proteins

sAIM: To examine the serological response of patients with upper gastrointestinal diseases and Helicobocter pylon (Hpy/on)infection to two Hpyloriouter membrane proteins (OMPs) (Mr18 000 and Mr26 000) acquired by gene recombinant technique, and to determine the diagnostic significance of serological tests derived from these OMPs.METHODS: Recombinant vectors encoding the two Hpylori OMPs were used to transform and express in BL21 (DE3) E.coli After purification with NP-NTA agarose resin, colloid gold kits were prepared with purified recombinant proteins to detect H pyloH infection and H pylori-associated diseases by the immunity-marker technology. We selected 150 patients with Hpy/on‘infection and digestive symptoms wibhout previous treabnent, including chronic gastritis (n = 60), duodenal ulcer (n = 30), gastric ulcer (n = 30), and gastric cancer (n=30).As controls, 33 Hpylori-negative healthy volunteers were also recruited. Serum samples were collected from all subjects, and the antibodies to specific proteins of Hpylori were tested with the colloid gold test kits. The sensitivity,specificity and accuracy of the colloid gold tests were evaluated, by using the combination of standard diagnostic methods (^13C urea breath test and bacteria culture) and classic enzyme-linked immunosorbent assay (ELISA) as reference.RESULTS: After purification with Ni^2+-NTA agarose resin,the purity of recombinant fusion proteins was about 95%.The recombinant fusion proteins were recognized by the specific monodonal antibodies against bhe two Hpy/oriOMPs,as demonstrated by the ELISA. Of the 150 serum samples from patients infected with Hpy/oH 141 (94.0%) responded positively to the recombinant protein with Mr26 000, while the seropositive rates were 95.0%, 96.7%, 96.7% and 90.0% for patients with H pylori-associated chronic gastritis,duodenal ulcer, gastric ulcer, and gastric cancer respectively.The sensitivity, specificity, and accuracy of the colloid gold kit with Mr26 000 protein were 94.0%, 97.0%, and 94.5%,respe.ctively. Compared with the classic ELISA, bacteria culture and ^13C urea breath test results in detecting Hpyloriinfection, there was no significant difference (P>O.O5). For the colloid gold kit with Mr18 000, the seropositive rates were 52.0%, 40.0%, 40.0%, 53.3% and 86.7%, respectively,in Hpylori-infected palJents, and bhose wibh Hpylori-associated chronic gastritis, duodenal ulcer, gastric ulcer, and gastric cancer. There was a significant difference (P<0.05) in seropositivity between patient with gastric cancer (86.7%) and those with other diseases (43.3%).CONCLUSION: The two colloid gold kits derived from the recombinant OMPs are useful tools either for detecting Hpyloriinfection, or for, predicting Hpylori-associated gastric malignancy.