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Diversity of Helicobacter pylori isolates in expression of antigens and induction of antibodies

World Journal of Gastroenterology 2008年第30期14卷 4816-4822页

作者： Ren-Xian Tang Dong-Jiao Luo ai-hua sun Jie Yan Department of Medical Microbiology and Parasitology Xuzhou Medical College Xuzhou 221009 Jiangsu Province China Qianjiang College Hangzhou Normal University Hangzhou 310018 Zhejiang Province China Faculty of Basic Medicine Zhejiang Medical College Hangzhou 310053 Zhejiang Province China Department of Medical Microbiology and Parasitology College of Medicine Zhejiang University Hangzhou 310053 Zhejiang Province China

aiM: To obtain evidence for selection of antigens used in genetically engineered vaccine against Helicobacter pylori (H pylori). METHODS: Enzyme linked immunoabsorbent assay (ELISA) was established on the basis of recombinant protein antigens rUreB, rHpaA, rVacA, rCagA1, rNapA, rFlaA and rFlaB of H pylori to detect expression rates of the antigens in bacterial isolates as well as positive rates of the antibodies in sera from H pylori-infected patients. PCR was applied to the detection of carrying rates of the genes encoding antigens in the isolates. RESULTS: The outputs of rUreB, rHpaA, rVacA, rCagA1, rNapA, rFlaA and rFlaB were approximately 35%, 32%, 15%, 23%, 56%, 25% and 20% of the total bacterial proteins, respectively. One hundred and fifty-one strains of H pylori were isolated from 347 biopsy specimens of chronic gastritis, peptic ulcer or gastric adenocarcinoma, with a positive rate of 43.5%. All of the isolates expressed UreB, HpaA, FlaA and FlaB while 52.3%, 92.1% and 93.4% of the isolates expressed VacA, CagA and NapA, respectively. In the sera of 151 H pylori-infected patients, the positive ratesof IgG antibodies against UreB, HpaA, VacA, CagA, NapA, FlaA and FlaB were 100%, 87.4%, 43%, 71.5%, 89.4%, 84.8% and 79.5%, respectively. Furthermore, the expression frequencies of VacA and NapA were found to be relative to the severity of gastric diseases (P = 0.016 and P ainst H pylori followed by NapA or HpaA.

关键词： Helicobacter pylori Major protein antigens Expression frequency Antibody levels

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CagA^+ *** Induces Akt1 Phosphorylation and Inhibits Transcription of p21^(WAF1/CIP1) and p27^(KIP1) via PI3K/Akt1 Pathway

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Biomedical and Environmental Sciences 2010年第4期23卷 273-278页

作者： SHU-PING LI XUE-JUN CHEN ai-hua sun JIN-FANG ZHAO JIE YAN State Key Laboratory for Diagnosis and Treatment of Infectious Diseases the First Affiliated Hospital of Zhejiang University Hangzhou 310003 Zhejiang China Department of Clinical Laboratory The Children's Hospital of Zhejiang University Hangzhou 310003 Zhejiang China Faculty of Basic Medicine Zhejiang Medical College Hangzhou 310053 Zhejiang China Department of Medical Microbiology and Parasitology Medical College of Zhejiang University Hangzhou 310058 Zhejiang China

Objective Cytotoxin-associated protein （CagA） of H. pylori has been confirmed to be closely associated with gastric inflammation and tumorigenesis, but the mechanism behind it is little understood. In this study, we try to determine roles of CagA＋ strain in activating PI3K/Akt1 signaling pathway, and affecting expression of p21WAF1/CIP1 and p27KIP1, and also in releasing IL-8 in host cells. Methods Akt1 phosphorylation and IL-8 levels of CagA＋ and CagAˉ strain infected AGS cells were detected by ELISAs. Two quantitative RT-PCRs were established to measure p21WAF1/CIP1 and p27KIP1 mRNA levels in the CagA＋ and CagAˉ strain infected cells. LY294002, an inhibitor of PI3K/Akt pathway, was used to define effect of the pathway in IL-8 release. Results CagA＋ strain could induce an obvious elevation of Akt1 phosphorylation in the infected AGS cells while CagAˉ strain failed to do so. The CagA＋ H. pylori strain infected AGS cells showed significant drops both in p21WAF1/CIP1 and p27KIP1 mRNA levels, whereas the CagAˉ H. pylori strain caused a remarkable increase in p21WAF1/CIP1 mRNA without affecting p27KIP1 gene transcription in the AGS cells. Both the CagA＋ and CagAˉ H. pylori strains enabled AGS cells to produce close elevated levels of IL-8, and the LY294002 block resulted in unexpected elevations of IL-8 levels. Conclusion CagA can activate PI3K/Akt1 pathway that plays an inhibitory role in IL-8 release in H. pylori infected AGS cells. Activation of PI3K/Akt1 pathway and subsequent negative regulation of p21^WAF1/CIP1 and p27^KIP1 expression might be involved in CagA-associated carcinogenesis.

关键词： Helicobater pylori CagA PI3K Akt1 p21^WAF1/CIP1 p27^KIP1 IL-8

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Rapid Detection of rpoB Gene Mutations in Rif-resistant *** Isolates by Oligonucleotide Microarray

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Biomedical and Environmental Sciences 2009年第3期22卷 253-258页

作者： ai-hua sun XING-LI FAN LI-WEI LI LI-FANG WANG WEN-YING AN JIE YAN Department of Basic Medicine Zhejiang Medical College Hangzhou 310053 Zhejiang China Department of Medical Microbiology and Parasitology Zhejiang University Hangzhou 310058 Zhejiang China School of lnternational Stud3 Zhejiang University Hangzhou 310058 Zhejiang China

Objective To detect the specific mutations in rpoB gene of Mycobacterium tuberculosis by oligonucleotide microarray. Methods Four wild-type and 8 mutant probes were used to detect rifampin resistant strains. Target DNA of M. tuberculosis was amplified by PCR, hybridized and scanned. Direct sequencing was performed to verify the results of oligonucleotide microarray Results Of the 102 rifampin-resistant strains 98 （96.1%） had mutations in the rpoB genes. Conclusion Oligonucleotide microarray with mutation-specific probes is a reliable and useful tool for the rapid and accurate diagnosis of rifampin resistance in M. tuberculosis isolates.

关键词： Mycobacterium tuberculosis Rifampin resistance rpoB gene / site mutation Oligonucleotide microarray/detection

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Usefulness of interferon-γrelease assay for the diagnosis of sputum smear-negative pulmonary and extra-pulmonary TB in Zhejiang Province,China

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Infectious Diseases of Poverty 2017年第1期6卷 1074-1078页

作者： Lei Ji Yong-Liang Lou Zhong-Xiu Wu Jin-Qin Jiang Xing-Li Fan Li-Fang Wang Xiao-Xiang Liu Peng Du Jie Yan ai-hua sun Department of Basic Medicine Hangzhou Medical CollegeHangzhou 310053ZhejiangPeople’s Republic of China School of Laboratory Medical Science and Life Science Wenzhou Medical UniversityWenzhou 325035ZhejiangPeople’s Republic of China Zhejiang Blood Center Hangzhou 310006ZhejiangPeople’s Republic of China Division of Basic Medical Microbiology State Key Laboratory for Diagnosis and Treatment of Infectious DiseasesFirst Affiliated Hospital of Zhejiang UniversityHangzhou 310003ZhejiangPeople’s Republic of China Department of Medical Microbiology and Parasitology College of MedicineZhejiang UniversityHangzhou 310058ZhejiangPeople’s Republic of China

Background:Quick diagnosis of smear-negative pulmonary tuberculosis(TB)and extra-pulmonary TB are urgently needed in clinical *** research aims to investigate the usefulness of the interferon-γrelease assay(IGRA)for the diagnosis of smear-negative pulmonary and extra-pulmonary ***:We performed TB antibody and TB-IGRA tests on 389 pulmonary TB patients(including 120 smear-positive pulmonary TB patients and 269 smear-negative pulmonary TB patients),113 extra-pulmonary TB patients,81 patients with other pulmonary diseases and 100 healthy *** samples for the TB-Ab test and the TB-IGRA were collected,processed,and interpreted according to the manufacturer’s ***:The detection ratio of smear-positive pulmonary TB patients and smear-negative pulmonary TB patients were 90.8%(109 of 120)and 89.6%(241 of 269),*** was no statistically significant difference of its performance between these two sample sets(P>0.05).The detection ratio of positive TB patients and extra-pulmonary TB patients were 90.0%(350 of 389)and 87.6%(99 of 113),respectively,which was not significantly different(P>0.05).Conclusions:In this work,the total detection ratio using TB-IGRA was 89.4%,therefore TB-IGRA has diagnostic values in smear-negative pulmonary TB and extra-pulmonary TB diagnosis.

关键词： Interferon-gamma release assay Smear-negative pulmonary TB Extra-pulmonary TB

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Diffusion in vivo and excretion from urine by transcytosis and propagation in mouse fibroblasts of ***

Diffusion in vivo and excretion from urine by transcytosis a...

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2017年浙江省微生物学会青年论坛——微生物与健康专题研讨会

作者： Yang Li ai-hua sun Wei-Lin Hu Xu'ai Lin 严杰 Zhejiang University Hangzhou Medical College 浙江大学

[Background]The animal hosts of *** can persistently excrete leptospires in urine and the leptospires in leptospirosis patients usually diffuse from blood stream into internal organs,but the mechanisms of *** to pass through blood vessel and renal tubule have not been reported.[Methods]Confocal microscopy,Transwell test,transfection and siRNA interference were used to determine the transcytosis of *** strain Lai through human/mouse HUVEC/EOMA blood vessel endothelial cells or HK-2/TCMK-1 renal tubular epithelial cells and propagation of the spirochete in human BJ or mouse NIH3T3 fibroblasts.[Results]The spirochete could invade into all the tested non-phagocytotic cells to form intracellular Lep-vesicles but this invasion *** blocked by a receptor-mediated endocytosis inhibitor(MDC).The Lep-vesicles did not fuse with lysosomes but excreted by transcytosis through HUVEC/EOMA or HK-2/TCMK-1 cells with the formation in turn of Rab5-recruited Lep-early-endosome,Rab 11-recruited Lep-recycling-endosome,Sec15/Sec3-recruited Lep-exocyst complex and VAMP2/SYN1-recruited Lep-SNARE *** intracellular Lep-propagation was only found in NIH3T3 fibroblasts and the leptospires in the mouse fibroblasts was excreted with the same manner as described above.[Conclusions]*** utilizes Rab-based endocytic recycling system and Secl5/Sec3-VAMP2/SYN1-based vesicular transport system for transcytosis which help us to understand the mechanisms of diffusion in leptospirosis patients and excretion in host animal urine of ***.

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Prognostic value and predictive model of tumor markers in stageⅠtoⅢgastric cancer patients

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World Journal of Clinical Oncology 2024年第8期15卷 1033-1047页

作者： ai-hua sun Xin-Yu Zhang Yang-Yang huang Lei Chen Qing Wang Xiao-Cong Jiang Department of Radiotherapy Oncology Huizhou Municipal Central HospitalHuizhou 516001Guangdong ProvinceChina Department of Hepatobiliary Surgery Fujian Provincial HospitalFuzhou 350001China Department of General Surgery Xiang'an Hospital of Xiamen UniversityXiamen 361102Fujian ProvinceChina

BACKGROUND Preoperative serum tumor markers have been widely used in the diagnosis and treatment of gastric cancer ***,few studies have evaluated the prognosis of gastric cancer patients by establishing statistical models with multiple serum tumor *** To explore the prognostic value and predictive model of tumor markers in stage I and III gastric cancer *** From October 2018 to April 2020,a total of 1236 patients with stage I to III gastric cancer after surgery were included in our *** relationship between serum tumor markers and clinical and pathological data were *** established a statistical model to predict the prognosis of gastric cancer based on the results of COX regression *** survival(OS)was also compared across different stages of gastric *** The deadline for follow-up was May 31,2023.A total of 1236 patients were included in our *** analysis found that age,clinical stage,T and N stage,tumor location,differentiation,Borrmann type,size,and four serum tumor markers were prognostic factors of OS(Pained from the statistical model,we found that patients with high scores had poorer survival time(P<0.05).Furthermore,in stage I patients,the 3-year OS for scores 0-3 ranged from 96.85%,95%,85%,and 80%.In stage II patients,the 3-year OS for scores 0-4 were 88.6%,76.5%,90.5%,65.5%and 60%.For stage III patients,3-year OS for scores 0-6 were 70.9%,68.3%,64.1%,50.9%,38.4%,18.5%and 5.2%.We also analyzed the mean survival of patients with different *** stage I patients,the mean OS was 55.980 *** stage II,the mean OS was 51.550 *** mean OS for stage III was 39.422 *** Our statistical model can effectively predict the prognosis of gastric cancer patients.

关键词： Gastric cancer Tumor marker Prognosis Overall survival Model

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A FasL-like outer membrane protein and lipopolysaccharide of *** induce macrophage apoptosis through Fas/FasL-caspase-8/-3 pathway

A FasL-like outer membrane protein and lipopolysaccharide of...

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2017年浙江省微生物学会青年论坛——微生物与健康专题研讨会

作者：杜蓬 Shi-Jun Li Wei-Lin Hu Kai-Xuan Li Xu'ai Lin ai-hua sun Jie Yan Zhejiang University 杭州医学院 Guizhou Provincial Centre for Disease Control and Prevention Hangzhou Medical College

[Background]Leptospira interrogans has been reported to induce apoptosis of macrophages during infection,but its molecular basis remains ***:Outer membrane proteins and lipopolysaccharide were extracted from *** strain Lai(Lep-OMP and Lep-LPS).Fas-captured Lep-OMP was identified by nanoLC-MS/***-binding ability of Lep-OMP was determined by surface plasmon resonance(SPR) and isothermal titration calorimetry(ITC).Lep-OMP-or Lep-LPS-induced mouse J774 A.1 and human THP-1 macrophage apoptosis,caspase-3/-8 activation,TLR2/4-dependent JNK/p38 MAPK/ERK phosphorylation,c-Jun/ATF2/CHOP translocation,Fas/FasL expression and translocation were determined by flow cytometry,Western Blotting,confocal microscopy and qRT-PCR.[Results]Lep-OMP047,the product of leptospiral LB047 gene,is the unique Fas-captured ***-OMP047 presented a powerful Fas-binding ability with the SPR and ITC K values of 5.20×10-2.84×10.10μg rLep-OMP047 or 1 ug Lep-LPS caused 33.6%and 22.1%or 21.9%and 15.8%early-apoptotic ratios in J774 A.1 and THP-1 macrophages,but Fas-IgG and caspase-8/-3 inhibitors blocked the Lep-OMP047-induced macrophage apoptosis while JNK/p38 MAPK inhibitors and TLR2-depletion inhibited the Lep-LPS-induced ***-LPS caused the TLR2-dependent JNK/p38 MAPK activation by phosphorylation and C-Jun/ATF2 nuclear translocation that involved in the Fas/FasL expression increase and membrane translocation.[Conclusions]The FasL-like OMP and LPS of *** synergistically induce the apoptosis of mouse/human macrophages through Fas/FasL-caspase-8/-3 pathway.

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Factors influencing engraftment in HLA-haploidentical/mismatch related transplantation with combined granulocyte-colony stimulating factor-mobilized peripheral blood and bone marrow for patients with Leukemia

Factors influencing engraftment in HLA-haploidentical/mismat...

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第13届全国实验血液学会议

作者： Cheng Zhang,Xi Zhang,Xing-hua Chen,Li Gao,Lei Gao,Pei-Yan Kong,Xian-Gui Peng,ai-hua sun,Qing-Yu Wang Department of Hematology,Xinqiao Hospital,the Third Military Medical University,Chongqing,400037

＜正＞Unmanipulated HLA-haploidentical/mismatch related transplantation with combined granulocyte-colony stimulating factor-mobilized peripheral blood stem cells （G-PBSCs） and granulocyte-colony stimulating factormobilized bone marrow（G-BM） has been developed as an alternative transplantation strategy for patients without an HLA-matched *** this transplantation setting, factors associated with hematopoietic recovery have not been defined *** aim of this study was to investigate the factors influencing the engraftment in this transplantation for patients with *** study group comprised 104 patients with leukemia who undergoing this transplantation at a single institution between 2005 and *** correlating with neutrophil and platelet engraftment post -transplantation were analyzed

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Regulation of acute graft-versus-host disease by human umbilical cord blood derived stromal cells in haploidentical stem cell transplantation in mice through very late activation antigen-4

Regulation of acute graft-versus-host disease by human umbil...

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第13届全国实验血液学会议

作者： Cheng Zhang,Xing-hua Chen,Xi Zhang,Lei Gao,Li Gao,Pei-Yan Kong,Xian-Gui Peng,ai-hua sun,Qing-Yu Wang Department of Hematology,Xinqiao Hospital,the Third Military Medical University,Chongqing 400037

＜正＞Human umbilical cord blood derived stromal cells （hUCBDSCs）,a novel resource isolated from CD34+ cells by our laboratory,have been previously shown not only to be nonimmunogenic but also to exert an immunosuppressive effect on xenogenic T cells in vitro. Very late activation antigen-4（VLA-4） has been associated with graft-versus-host disease（GVHD）.This study aimed to investigate the possible mechanism by which hUCBDSCs regulate acute GVHD（aGVHD） in haploidentical stem cell transplantation in *** of VLA-4 was explored by flow cytometry in spleen lymphocytes of donor mice cocultured with hUCBDSCs and in haploidentical stem cell transplantation in mice with *** hUCBDSCs and human bone marrow stromal cells（hBMSCs） elicited decreased lymphocyte expression of VLA-4,but this decrease was stronger with hUCBDSCs than with hBMSCs（p＜0.05）. Cotransplantation of bone marrow with hUCBDSCs significantly decreased the expression of VLA-4 compared with control mice（p＜0.05）.A significant reduction of VLA-4 labeling in the target organs of GVHD was evident in haploidentical mice cotransplanted with hUCBDSCs. Our study shows that hUCBDSCs may protect mouse recipients of haploidentical stem cell transplantation from aGVHD via downregulating the expression of VLA-4.

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技能表现类项群运动员流畅心理状态诱发因素及其可控性研究

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天津体育学院学报 2004年第1期19卷 27-30页

作者：胡咏梅孙爱华孙延林吉承恕天津体育学院天津300381

通过访谈25名技能表现类项群的杰出运动员,总结出此项群运动员流畅心理状态的九个诱发因素,按重要性依次排序:适宜的身体准备、赛前和赛中的计划准备、自信心和积极的态度、赛前达到适宜的唤醒水平、良好的比赛动机、赛中感觉好、集中... 详细信息

通过访谈25名技能表现类项群的杰出运动员,总结出此项群运动员流畅心理状态的九个诱发因素,按重要性依次排序:适宜的身体准备、赛前和赛中的计划准备、自信心和积极的态度、赛前达到适宜的唤醒水平、良好的比赛动机、赛中感觉好、集中注意力、适宜的比赛情境、教练和团队的配合。其中,排序前5个的是主要诱发因素。流畅心理状态可控性的调查结果显示:75%的运动员认为流畅状态是可以控制的,在69条诱发因素的原始条目中,能够被控制的条目占总数的73.3%,通过赛前和赛中的心理调控帮助运动员进入并保持流畅状态是完全可行的。教练员和运动心理学研究者可以依据表现类项群9个诱发因素制定相应的心理训练方案。

关键词：技能表现类流畅心理状态诱发因素可控性

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