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排序方式: 共有165条查询结果,搜索用时 15 毫秒
1.
目的 克隆幽门螺杆菌(Helicobacter pylori,HP)过氧化氢酶(katA)基因并在大肠杆菌中进行表达。方法 采用PCR扩增幽门螺杆菌katA全长基因,将其克隆入pET-11c载体中,经测序证实后,在大肠杆菌中进行表达,产物用Western blot检测其抗原性并进行N末端氨基酸的测序。结果 幽门螺杆菌katA基因全长1 518bp,编码氨基酸505个。在BL21(DE3)中的表达量约占细菌总蛋白的24.9%,表达产物经SDS-PAGE显示其相对分子质量与软件预测结果 58 000相符,N末端5个氨基酸测序结果与Hp中天然的katA完全一致,经Western blot检测可被 Hp全菌抗血清识别。结论 katA能在大肠杆菌中进行高效表达,具有良好的免疫反应性,可望为研究 Hp的致病机理、实验诊断及亚单位疫苗等提供充足的katA原材料。 相似文献
2.
从包涵体中纯化重组人幽门螺杆菌尿素酶B亚单位 总被引:3,自引:0,他引:3
目的 建立一种有效的方法,从包涵体中纯化重组人幽门螺杆菌尿素酶B亚单位(rUreB)。方法重组大肠杆菌发酵后,表达的rUreB包涵体经洗涤、变性、复性,采用Q Sepharose Performance阴离子交换层析和Pheny1 Sepharose High Performance疏水层析分离纯化。使用 SDS-PAGE和HPLC检测纯度,选用 ELISA和 Western-bloning对纯化蛋白的生物学活性进行鉴定。结果 rUreB包涵体经洗涤和溶解后,rUreB的纯度>70%。包涵体溶解液经阴离子交换层析和疏水层析后纯度超过95%。rUreB纯品具有良好的生物学活性。结论 建立了从包涵体中获得高纯度的rUreB的工艺,为进一步的研究打下了基础。 相似文献
3.
目的建立并评价幽门螺杆菌(Hp)尿素酶B亚单位(UreB)特异性抗体的检测方法。方法应用纯化的重组HpUreB作为包被抗原,建立检测HpUreB特异性抗体的ELISA间接法。以Hp抗体Western blot检测作为“金标准”,评价所建立方法的真实性和可靠性。结果检测血清特异性IgG的灵敏度为100·0%,特异度为97·7%,阳性预测值为97·2%,阴性预测值为100·0%,约登指数为0·977,符合率为98·7%,试验的一致率为98·5%;检测血清中总的特异性Ig灵敏度为97·1%,特异度为95·3%,阳性预测值为94·4%,阴性预测值为97·6%,约登指数为0·925,符合率为96·2%,试验的一致率为97·8%;检测唾液中特异性sIgA的灵敏度为96·2%,特异度为94·5%,阳性预测值为89·3%,阴性预测值为98·1%,约登指数为0·907,符合率为95·1%,试验的一致率为97·5%;检测粪便标本中的特异性sIgA的灵敏度为92·0%,特异度为90·2%,阳性预测值为85·2%,阴性预测值为94·9%,约登指数为0·822,符合率为90·9%,试验的一致率为98·6%,CV值均小于15%。结论该检测方法真实性、可靠性、重复性良好,能满足临床标本检测的要求。 相似文献
4.
将以具有链状高分子结构的柠檬酸铋为模板和前体合成出本身具有抑菌活性的含铋纳米材料,并以此为载体,通过吸附、填充、化学结合等方法与药物小分子或抗生素形成复合材料.研究含铋纳米材料和小分子复合过程的相互作用.评价含铋纳米材料在复合前后对幽门螺杆菌生长的抑制活性的变化.初步研究在模拟生理条件下,纳米材料分解和释放药物的过程,为研究开发新的治疗消化性溃疡疾病的药物提供新的思路. 相似文献
5.
Bernarde C Khoder G Lehours P Burucoa C Fauchère JL Delchier JC Mégraud F Atanassov C 《Proteomics. Clinical applications》2009,3(6):672-681
To date no reliable diagnostic method exists to predict, among the very large and clinically heterogeneous group of Helicobacter pylori‐infected patients, the extremely small group at risk for developing low‐grade gastric MALT lymphoma (LG‐MALT). Search of proteomic biomarkers holds promise for the classification of the H. pylori strains with regard to this severe clinical outcome. In the present study 69 H. pylori strains isolated from patients with two different H. pylori‐associated diseases, duodenal ulcer (DU, n=29) and LG‐MALT (n=40) were used. Protein expression patterns of the strains were analyzed by using the high‐throughput methodology SELDI. Selected proteins were purified by means of chromatographic and electrophoretic methods in view of further sequencing by LC‐MS/MS. Univariate analysis (Mann–Whitney test) of the protein expression patterns generated nine significant biomarkers that can discriminate between H. pylori strains from patients with DU and LG‐MALT. These biomarkers are of low molecular weight, ranging from 6 to 26.6 kDa. Among them, two are overexpressed in LG‐MALT strains and seven – in DU strains. Two biomarker proteins, one overexpressed in LG‐MALT strains (13.2 kDa) and another one – overexpressed in DU strains (26.6 kDa), were purified to homogeneity and identified by using LC‐MS/MS as a 50S ribosomal protein L7/L12 and a urease subunit, respectively. These biomarkers can be included in novel protein arrays for the differential diagnosis of H. pylori‐associated clinical outcomes. 相似文献
6.
Lornella Seeneevassen Emilie Bessde Francis Mgraud Philippe Lehours Pierre Dubus Christine Varon 《International journal of molecular sciences》2021,22(7)
Gastric cancer’s bad incidence, prognosis, cellular and molecular heterogeneity amongst others make this disease a major health issue worldwide. Understanding this affliction is a priority for proper patients’ management and for the development of efficient therapeutical strategies. This review gives an overview of major scientific advances, made during the past 5-years, to improve the comprehension of gastric adenocarcinoma. A focus was made on the different actors of gastric carcinogenesis, including, Helicobacter pylori cancer stem cells, tumour microenvironment and microbiota. New and recent potential biomarkers were assessed as well as emerging therapeutical strategies involving cancer stem cells targeting as well as immunotherapy. Finally, recent experimental models to study this highly complex disease were discussed, highlighting the importance of gastric cancer understanding in the hard-fought struggle against cancer relapse, metastasis and bad prognosis. 相似文献
7.
Zhongming Ge Lili Ge Sureshkumar Muthupalani Yan Feng James G. Fox 《International journal of molecular sciences》2020,21(23)
The prevalence of gastric Helicobacter pylori (Hp) infection is ~50% of the world population. However, how Hp infection influences inflammatory bowel disease in humans is not fully defined. In this study, we examined whether co-infection with Hp influenced Helicobacter hepaticus (Hh)–induced intestinal pathology in Rag2−/− mice. Rag2−/− mice of both sexes were infected with Hh, of which a subgroup was followed by infection with Hp two weeks later. Co-infected males, but not females, had significantly higher total colitis index scores in the colon at both 10 and 21 weeks post-Hh infection (WPI) and developed more severe dysplasia at 21 WPI compared with mono-Hh males. There were no significant differences in colonization levels of gastric Hp and colonic Hh between sexes or time-points. In addition, mRNA levels of colonic Il-1β, Ifnγ, Tnfα, Il-17A, Il-17F, Il-18, and Il-23, which play important roles in the development and function of proinflammatory innate lymphoid cell groups 1 and 3, were significantly up-regulated in the dually infected males compared with mono-Hh males at 21 WPI. These data suggest that concomitant Hp infection enhances the inflammatory responses in the colon of-Hh-infected Rag2−/− males, which results in more severe colitis and dysplasia. 相似文献
8.
Jacky Lu Kathryn P. Haley Jamisha D. Francis Miriam A. Guevara Ryan S. Doster Kelly M. Craft Rebecca E. Moore Schuyler A. Chambers Alberto G. Delgado Maria Blanca Piazuelo Steven M. Damo Dr. Steven D. Townsend Dr. Jennifer A. Gaddy 《Chembiochem : a European journal of chemical biology》2021,22(18):2783-2790
Chronic infection with Helicobacter pylori increases risk of gastric diseases including gastric cancer. Despite development of a robust immune response, H. pylori persists in the gastric niche. Progression of gastric inflammation to serious disease outcomes is associated with infection with H. pylori strains which encode the cag Type IV Secretion System (cag T4SS). The cag T4SS is responsible for translocating the oncogenic protein CagA into host cells and inducing pro-inflammatory and carcinogenic signaling cascades. Our previous work demonstrated that nutrient iron modulates the activity of the T4SS and biogenesis of T4SS pili. In response to H. pylori infection, the host produces a variety of antimicrobial molecules, including the iron-binding glycoprotein, lactoferrin. Our work shows that apo-lactoferrin exerts antimicrobial activity against H. pylori under iron-limited conditions, while holo-lactoferrin enhances bacterial growth. Culturing H. pylori in the presence of holo-lactoferrin prior to co-culture with gastric epithelial cells, results in repression of the cag T4SS activity. Concomitantly, a decrease in biogenesis of cag T4SS pili at the host-pathogen interface was observed under these culture conditions by high-resolution electron microscopy analyses. Taken together, these results indicate that acquisition of alternate sources of nutrient iron plays a role in regulating the pro-inflammatory activity of a bacterial secretion system and present novel therapeutic targets for the treatment of H. pylori-related disease. 相似文献
9.
Sadia Alvi Aqeel Javeed Bushra Akhtar Ali Sharif Muhammad Furqan Akhtar 《International Journal of Food Properties》2017,20(10):2215-2222
Human physiology normally contains pathogenic and non-pathogenic microorganisms in the gastrointestinal flora. Disturbance of these microorganism balance results in the formation of infection. Extensive use of antibiotics for cure of these disturbances like Helicobacter pylori (H. pylori) infection leads to patient discomfort and associated side-effects. There is a need to adopt adjunct or alternative approach in order to minimize such conditions. Probiotics is one of the potential therapies to cure gastrointestinal discomforts especially associated with H. pylori. It competes through non-immune and immune systems. This review article concludes that probiotics are used to eradicate the infection at increased rate, and decreased associated side-effects are caused by triple therapy. A proper evaluation of these probiotics is demanded before their use in future as a commercial product. Furthermore, their effect on immune system requires more research work so that their usage for other chronic disorders can also be considered. 相似文献
10.
Claudia Zagami Diana Papp Alice Anna Daddi Francesco Boccellato 《International journal of molecular sciences》2022,23(7)
The adult gastric mucosa is characterised by deep invaginations of the epithelium called glands. These tissue architectural elements are maintained with the contribution of morphogen signals. Morphogens are expressed in specific areas of the tissue, and their diffusion generates gradients in the microenvironment. Cells at different positions in the gland sense a specific combination of signals that instruct them to differentiate, proliferate, regenerate, or migrate. Differentiated cells perform specific functions involved in digestion, such as the production of protective mucus and the secretion of digestive enzymes or gastric acid. Biopsies from gastric precancerous conditions usually display tissue aberrations and change the shape of the glands. Alteration of the morphogen signalling microenvironment is likely to underlie those conditions. Furthermore, genes involved in morphogen signalling pathways are found to be frequently mutated in gastric cancer. We summarise the most recent findings regarding alterations of morphogen signalling during gastric carcinogenesis, and we highlight the new stem cell technologies that are improving our understanding of the regulation of human tissue shape. 相似文献