|
|
|
题名
|
作者
|
年代
|
出处
|
被引量
|
| 1 | Construction of recombinant attenuated Salmonella typhimurium DNA vaccine expressing H pylori ureB and IL-2显示文摘AIM: To construct a recombinant live attenuated Salm-onella typhimurium DNA vaccine encoding H pylori ureB gene and mouse IL-2 gene and to detect its immunogenicity in vitro and in vivo. METHODS: H pylori ureB and mouse IL-2 gene fragments were amplified by polymerase chain reaction (PCR) and cloned into pUCmT vector. DNA sequence of the amplified ureB and IL-2 genes was assayed, then cloned into the eukaryotic expression vector pIRES through enzyme digestion and ligation reactions resulting in pIRES-ureB and pIRES-ureB-IL-2. The recombinant plasmids were used to transform competent E. coli DH5α, and the positive clones were screened by PCR and restriction enzyme digestion. Then, the recombinant pIRES-ureB and pIRES-ureB-IL-2 were used to transform LB5000 and the recombinant plasmids extracted from LB5000 were finally introduced into the final host SL7207. After that, recombinant strains were grown in vitro repeatedly. In order to detect the immunogenicity of the vaccine in vitro, pIRES-ureB and pIRES-ureB-IL-2 were transfected to COS-7 cells using LipofectamineTM2000, the immunogenicity of expressed UreB and IL-2 proteins was assayed with SDS-PAGE and Western blot. C57BL/6 mice were orally immunized with 1 × 108 recombinant attenuated Salmonella typhimurium DNA vaccine. Four weeks after vaccination, mice were challenged with 1 × 107 CFU of live H pylori SS1. Mice were sacrificed and the stomach was isolated for examination of H pylori 4 wk post-challenge. RESULTS: The 1700 base pair ureB gene fragment amplified from the genomic DNA was consistent with the sequence of H pylori ureB by sequence analysis. The amplified 510 base pair fragment was consistentwith the sequence of mouse IL-2 in gene bank. It was confirmed by PCR and restriction enzyme digestion that H pylori ureB and mouse IL-2 genes were inserted into the eukaryotic expression vector pIRES. The experiments in vitro showed that stable recombinant live attenuated Salmonella typhimurium DNA vaccine carrying ureB and IL-2 genes was successfully constructed and the specific strips of UreB and IL-2 expressed by recombinant plasmids were detected through Western blot. Study in vivo showed that the positive rate of rapid urease test of the immunized group including ureB and ureB-IL-2 was 37.5% and 12.5% respectively, and was significantly lower than that (100%) in the control group (P < 0.01). CONCLUSION: Recombinant attenuated Salmonella typhimurium DNA vaccine expressing UreB protein and IL-2 protein with immunogenicity can be constructed. It can protect mice against H pylori infection, which may help the development of a human-use H pylori DNA vaccine. | Can Xu Zhao-Shen Li Yi-Qi Du Yan-Fang Gong Hua Yang Bo Sun Jing Jin | 2007 | World Journal of Gastroenterology2007，13，6： | 10 |
| 2 | Helicobacterpylori virulence factors in duodenal ulceration:A primary cause or a secondary infection causing chronicity显示文摘感染不看十二指肠的溃疡的成比例地的高流行的来自有 Helicobacter pylori (H pylori ) 的高流行的国家的报告,建议 H pylori 不能是十二指肠的溃疡的一个原发性原因的可能性。这差异可能被变化在不同人口在毒力因素的流行解释,这被提出了。这篇论文的目的是决定出版文学是否支持这可能性。相关文学与 H pylori 感染和毒力因素的高、低的流行为国家独立被考察并且分析。尽管 H pylori 的剧毒的紧张是,更经常显著地与 H pylori 感染的低流行在国家比没有疾病与十二指肠溃疡在病人介绍在人口,在在十二指肠溃疡之间的毒力因素的流行没有差别,非-在许多国家的溃疡消化不良或正常题目,在两 H pylori 感染并且毒力因素的流行高的地方。在这些国家,毒力因素的存在不是预兆的临床的结果。为了在国家解释在毒力因素和十二指肠溃疡之间的协会,在 H pylori 流行是低的的地方,仅仅二份报纸被发现那几乎别把小支持给建议的平常的模型,也就是没有他们,有毒力因素的有机体是比那些更可能的开始十二指肠溃疡。我们提供建议毒力因素是更可能的防碍以前生产的溃疡愈合的一个其他的假设。毒力因素的存在仅仅在 H pylori 的流行是低的国家与十二指肠溃疡的流行相关。有很小的证据,毒力因素开始十二指肠的溃疡,但是他们可能与溃疡的失败有关愈合。 | FrankI Tovey Michael Hobsley John Holton | 2006 | World Journal of Gastroenterology2006，12，1： | 6 |
| 3 | 错配修复基因hMLH3在家族性胃癌中的突变显示文摘目的:检测错配修复基因hMLH3在家族性胃癌中的突变情况,以探讨hMLH3在家族性胃癌中的作用. 方法:采用聚合酶链反应(PCR)、变性高效液相色谱分析(DHPLC)和直接测序法,检查有遗传背景的16个胃癌家系(共84名成员)的错配修复基因hMLH3突变情况.胃癌家系选择参考以下标准:(1)至少连续两代;(2)至少2例胃癌患者;(3)至少有1例患者为其他患者的一级亲属;(4)至少有1 例患者50岁前被诊断出胃癌. 结果:所有样品的外显子均成功进行PCR扩增,DHPLC分析和基因测序.共在5个家系中发现5处错义突变(31.3%, 5/16),4处在外显子1,另外1处在外显子12.家系6中hMLH3的突变与胃癌发生呈现出较强的相关性,而另外几个家系中的突变情况与胃癌没有表现出明显的相关性.在散发性胃癌及正常对照中未发现突变的存在. 结论:hMLH3基因在家族性胃癌发生中可能为一低风险基因,他可能通过与其他基因互相叠加、共同起作用. | 赵成海 刘宏旭 卜献民 | 2004 | 世界华人消化杂志2004，12，5： | 5 |
| 4 | Prevalence of vacA, cagA and babA2 genes in Cuban Helicobacter pylori isolates显示文摘AIM: To investigate the prevalence of vacuolating cytotoxin (vacA), cytotoxin associated gene A (cagA) and blood adhesion binding antigen (babA2) genotypes of Helicobacter pylori (H pylori) isolates from Cuban dyspeptic patients. METHODS: DNA was extracted from H pylori-positive cultures taken from 130 dyspeptic patients. Genotyping was performed by PCR, using specifi c primers for vacA (s1, s2, m1, m2), cagA and babA2 genes. Endoscopic observations and histological examinations were used to determine patient pathologies. RESULTS: vacA alleles s1 , s2 , m1 and m2 were detected in 96 (73.8%), 34 (26.2%), 75 (57.7%) and 52 isolates (40%), respectively, while the cagA gene was detected in 95 isolates (73.2%). One hundred and seven isolates (82.3%) were babA2-positive. A signif icant correlation was observed between vacAs1m1 and cagA and between vacAs1m1 and babA2 genotypes (P < 0.001 and P < 0.05, respectively) and between babA2 genotype and cagA status (P < 0.05); but, no correlation was observed between vacAs1 and babA2 genotypes. Eighty fi ve (65.4%) and 73 (56.2%) strains were type 1 (vacAs1-cagA-positive) and 'triple- positive' (vacAs1-cagA-babA2-positive), respectively, and their presence was significantly associated with duodenal ulcer (P < 0.01 and P < 0.001, respectively). CONCLUSION: The distribution of the main virulence factors in the Cuban strains in this study resembled that of the Western-type strains, and the more virulent H pylori isolates were signifi cantly associated with duodenal ulcer, ulcer disease being the worst pathology observed in the group studied. | Lino E Torres Karelia Melián Arlenis Moreno Jordis Alonso Carlos A Sabatier Mayrín Hernández Ludisleydis Bermúdez Boris L Rodríguez | 2009 | World Journal of Gastroenterology2009，15，2： | 4 |
| 5 | 互联网时代的学习型城市建设显示文摘建设学习型城市是实施以人的现代化为核心的城镇化发展战略的重要内容。在当前的互联网时代,要从保护自主学习意识,注重学习能力的启发与提高,丰富学习资源,开展定制化教育服务,完善监管机制,保障互联网学习环境的洁净等方面入手,让互联网技术切实服务于学习型城市的建设工作之中。 | 黄霞 | 2014 | 中国成人教育2014，，4： | 4 |
| 6 | A new subtype of 3' region of cagA gene in Helicobacter pyloristrains isolated from Zhejiang Province in China显示文摘AIM: To isolate the subtypes of 3′ region of cagA gene in Helicobacter pylori (H pylon) strains from Zhejiang Province in China and to investigate their relations to H pylori-associated gastroduodenal diseases. METHODS: One hundred and thirty-seven H pylori clinical strains were isolated from the gastric mucosa specimens of 74 patients with chronic gastritis, 61 with peptic ulceration, and 2 with gastric cancer. Bacterial genomic DNA was extracted and 3′ region of cagA gene was amplified by polymerase chain reaction (PCR). Subtypes of 3′ region of cagA gene were determined by the size of PCR amplified segments. The sequences of the subtypes were analyzed by PCR-based sequencing. RESULTS: Of the 137 H pyloriisolates from Zhejiang Province, 132 (96.4%) yielded PCR products that could be classified into three groups of subtypes, named as subtypes Ⅰ, Ⅱ, and Ⅲ according to their sizes. The sizes of subtypes Ⅰ, Ⅱ, and Ⅲ were 648-650bp, 705-707bp, and 815bp, respectively. Among the 132 cagA-positive H pylori strains, 123(93.2%) belonged to the group of subtype Ⅰ, 6 (4.5%) presented subtype Ⅱ, 1(0.8%) was subtype Ⅲ, and 2(1.5%) presented subtypes Ⅰ and Ⅲ both. The primary structure of subtype Ⅰ was composed of 3 repeats of R1, 1 repeat of R2 and 1 repeat of R3. Subtype Ⅱ possessing 4 repeats of R1, 2 repeats of R2 and 1 repeat of R3 was a newly found type of 3′ region of cagA gene which had not been reported before. The primary structure of subtype Ⅲ consisted of 4 repeats of R1, Ⅰ repeat of R2 and 2 repeats of R3. Comparison of the sequences of subtype Ⅰ strains with the corresponding sequences deposited in GenBank, showed a similarity of 95.0% (94.0-96.1%) for nucleotide sequences and 95.9% (94.9-97.4%) for deduced amino acid sequences. Comparison of the sequences of subtype Ⅲ strains with the corresponding sequences deposited in GenBank, showed a similarity of 93.9% (90.8-96.9%) for nucleotide sequences and 93.2% (90.2-96.2%) for deduced amino acid sequences. Among subtype Ⅱ strains, the nucleotide and deduced amino acid sequences showed a similarity of 95.2% (94.1-96.5%) and 96.4% (93.8-97.9%),respectively. There were no statistical differences in the distribution of subtypes of 3′ region of cagA gene among different H pylori-associated gastroduodenal diseases (x^2=11.544, P>0.05). CONCLUSION: There are three subtypes (Ⅰ,Ⅱ, and Ⅲ) of 3′ region of cagA gene in Hpylori strains isolated from Zhejiang Province, and subtype Ⅰ is predominant. Subtype Ⅱ is a newly found subtype of 3′ region of cagA gene. The result of this study does not support the view that the subtypes of 3′ region of cagA gene in H pylori isolated from Zhejiang Province are correlated with the clinical outcomes of H pylori infection. | RanTao Ping-ChuFang Hai-YanLiu Yun-ShuiJiang JingChen | 2004 | World Journal of Gastroenterology2004，10，22： | 2 |
| 7 | 中医老年病学研究方法显示文摘中医老年病学应参考临床流行病学和循证医学的方法进行研究。在中医药研究领域引入建立在临床流行病学、统计学及现代信息技术应用基础上的系统评价和Meta分析方法,将有助于推动中医药学的发展。 | 宋至诚 邹勇 | 2014 | 中国中医药现代远程教育2014，12，5： | 1 |
| 8 | 移动接入网IP化显示文摘本文通过对移动网GSM/UMTS采用的体系结构的介绍,分析了移动接入网的现状及发展方向,并在工程实施的角度出发,详细描述了移动接入网络的具体步骤以及需要注意的问题。 | 郭思娟 | 2014 | 科技视界2014，，3： | 0 |
您所在的机构暂未开通此部分的功能，请联系管理员