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Establishment of a nested-ASP-PCR method to determine the clarithromycin resistance of Helicobacter pylori

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World Journal of Gastroenterology 2016年第25期22卷 5822-5830页

作者： Xiao-Feng Luo jian-Hua jiao Wen-Yue Zhang Han-ming Pu Bao-jin Qu Bing-Ya Yang min Hou min-jun ji Department of Pathogen Biology Nanjing Medical University Department of Gastroenterology First Affiliated Hospital of Nanjing Medical University Department of Gastroenterology Nanjing Qixia District Hospital

AIM: To investigate clarithromycin resistance positions 2142, 2143 and 2144 of the 23 Sr RNA gene in Helicobacter pylori(H. pylori) by nested-allele specific primer-polymerase chain reaction(nested-ASP-PCR).METHODS: The gastric tissue and saliva samples from 99 patients with positive results of the rapid urease test(RUT) were collected. The nested-ASP-PCR method was carried out with the external primers and inner allele-specific primers corresponding to the reference strain and clinical strains. Thirty gastric tissue and saliva samples were tested to determine the sensitivity of nested-ASP-PCR and ASP-PCR methods. Then, clarithromycin resistance was detected for 99 clinical samples by using different methods, including nestedASP-PCR, bacterial culture and disk diffusion. RESULTS: The nested-ASP-PCR method was successfully established to test the resistance mutation points 2142, 2143 and 2144 of the 23 SrR NA gene of H. pylori. Among 30 samples of gastric tissue and saliva, the H. pylori detection rate of nested-ASP-PCR was 90% and 83.33%, while the detection rate of ASP-PCR was just 63% and 56.67%. Especially in the saliva samples, nested-ASP-PCR showed much higher sensitivity in H. pylori detection and resistance mutation rates thanASP-PCR. In the 99 RUT-positive gastric tissue and saliva samples, the H. pylori-positive detection rate by nested-ASP-PCR was 87(87.88%) and 67(67.68%), in which there were 30 wild-type and 57 mutated strains in gastric tissue and 22 wild-type and 45 mutated strains in saliva. Genotype analysis showed that three-points mixed mutations were quite common, but different resistant strains were present in gastric mucosa and saliva. Compared to the high sensitivity shown by nested-ASP-PCR, the positive detection of bacterial culture with gastric tissue samples was 50 cases, in which only 26 drug-resistant strains were found through analyzing minimum inhibitory zone of clarithromycin. CONCLUSION: The nested-ASP-PCR assay showed higher detectio

关键词： Helicobacter pylori Nested-allele specific primer-polymerase chain reaction Rapid urease test Clarithromycin resistance Drug sensitivity testing

Characterization of CD4~+ T Cell Responses in Mice Infected with Schistosoma japonicum

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Acta Biochimica et Biophysica Sinica 2006年第5期38卷 327-334页

作者： min-jun ji Chuan SU Yong WANG Hai-Wei WU Xiao-Ping CAI Guang-Fu LI Xiang ZHU Xin-jun WANG Zhao-Song ZHANG Guan-Ling WU~* Department of Pathogen Biology,Nanjing Medical University,Nanjing 210029,China Department of Pathogen Biology Nanjing Medical UniversityNanjing 210029 China

To better understand the interaction between Schistosoma japonicum and its murine host,wecharacterized the immune response of CD4~+ T cells generated during an experimental *** infectionbased on different key aspects,from gene expression to cell *** oligonucleotide microarrayswere used to compare gene expression profiles of CD4~+ T cells from spleens of mice at 0,3,6 and 13weeks *** cytometry analysis was used to determine type 1 and type 2 cytokine-secretingCD4~+ T cells,to test apoptosis of CD4~+ T cells and to count CD4~+CD25~+ T cells,a kind of regulatorysubpopulation of CD4~+ T *** percentage of interleukin-4-producing CD4~+ T cells was found to bemuch higher than that of γ-interferon-producing cells,especially after stimulation with *** eggantigen,which was consistent with type 1 and type 2 cytokine gene expression in the *** also showed that *** induced the increased expression of Th2 response-related genes,whereassome transcripts related to the Th1 responsive pathway were *** cytometry analysis showeda marked increase in the apoptotic CD4~+ T cells from 6 weeks post-infection and in the ratio of CD4~+CD25~+to CD4~+ T cells in infected mice after 13 *** therefore concluded that experimental infection of micewith *** resulted in a Th2-skewed immune response,which was to a great extent monitored bythe immune regulatory network,including cytokine cross-modulation,cell apoptosis and the subpopulationof regulatory cells.

关键词： Schistosoma japonicum CD4~+ T cell immune deviation immune regulatory network

Identification of Immunodominant Th1-type T cell Epitopes from Schistosoma japonicum 28 kDa Glutathione-S-transferase,a Vaccine Candidate

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Acta Biochimica et Biophysica Sinica 2005年第11期37卷 751-758页

作者： Guang-Fu LI Yong WANG Zhao-Song ZHANG Xin-jun WANG min-jun ji Xiang ZHU Feng LIU Xiao-Ping CAI Hai-Wei WU Guan-Ling WU Department of Pathogenic Biology,Nanjing Medical University,Nanjing 210029,China Department of Pathogenic Biology Nanjing Medical UniversityNanjing 210029 China

Th1-type cytokines produced by the stimulation of Th1-type epitopes derived from definedschistosome-associated antigens are correlated with the development of resistance to the parasite *** mansoni 28 kDa glutathione-S-transferase(Sm28GST),a major detoxification enzyme,hasbeen recognized as a vaccine candidate and a phase Ⅱ clinical trial has been carried *** immunizedwith recombinant Schistosoma japonicum 28GST(Sj28GST)have shown immune protection against theparasite *** the present study,six candidate peptides(P1,P2,P3,P4,P7 and P8)from Sj28GSTwere predicted,using software,to be T cell epitopes,and peptides P5 and P6 were designed by extendingfive amino acids at the N-terminal and C-terminal of P1,*** peptide 190-211 aa in Sj28GSTcorresponding to the Th1-type epitope(190-211 aa)identified from Sm28GST was selected and named *** nine candidate peptides were synthesized or produced as the fusion protein with thioredoxin in thepET32c(+)/BL21(DE3)*** capacity to induce a Th1-type response in vitro was measured usinglymphocyte proliferation,cytokine detection experiments and flow *** results showed that P6(73-86 aa)generated the strongest stimulation effect on T cells among the nine candidate peptides,and drovethe highest level of IFN-γ and ***,P6 is a functional Th1-type T cell epitope that is differentfrom that in Sm28GST,and will be useful for the development of effective vaccines which can triggeracquired immunity against ***,our strategy of identifying the Th1-type epitope by acombination of software prediction and experimental confirmation provides a convenient and cost-savingalternative approach to previous methods.

关键词： Schistosoma japonicum epitope Sj28GST Th1 epitope IFN-γ

Gene Transcription Profile in Mice Vaccinated with Ultraviolet-attenuated Cercariae of Schistosoma japonicum Reveals Molecules Contributing to Elevated IFN-γ Levels

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Acta Biochimica et Biophysica Sinica 2005年第4期37卷 254-264页

作者： Xiang ZHU Zhao-Song ZHANG min-jun ji Hai-Wei WU Yong WANG Xiao-Ping CAI Lei ZHANG Shu-Ying HU Lin-Lin FU Feng LIU Chuan SU Guan-Ling WU Department of Pathogenic Biology,Nanjing Medical University.Nanjing 210029.China Department of Pathogenic Biology Nanjing Medical UniversityNanjing 210029 China

Vaccination with ultraviolet-attenuated cercariae of Schistosoma japonicum induced protectiveimmunity against challenge infection in experimental animal *** preliminary study on the transcrip-tion levels of IFN-γ and IL-4 in splenic CD4~+ T cells revealed that attenuated cercariae elicited predomi-nantly a Th1 response in mice at the early stage,whereas normal cercariae stimulated primarily Th2-dependent *** analysis on the gene profile of the skin-draining lymph nodes demonstratedthat the levels of IFN-γ were significantly higher in vaccinated mice than those in infected mice at day 4,7and 14 post-vaccination or ***,for IL-12 and IL-4,the potent inducers of Th1 andTh2 responses,respectively,as well as IL-10,there were no differences over the course of the experimentbetween the infected and vaccinated *** explore the underlying factors that may potentially contributeto elevated IFN-γ in vaccinated mice,the mRNA profiles of the skin-draining lymph nodes at day 4 post-exposure were compared using oligonucleotide *** the 847 probe sets with increased signalvalues,we focused on chemokines,cytokines and relevant receptors,which were validated by semi-quantitativeRT-PCR,A comprehensive understanding of the immune mechanisms of attenuated cercariae-inducedprotection may contribute to developing efficient vaccination strategies against ***,especiallyduring the early stare of infection.

关键词： Schistosoma japonicum attenuated cercariae oligonucleotide microarray IFN-γ chemokines

Dynamics of CD4~+CD25~+ T Cells in Spleens and Mesenteric Lymph Nodes of Mice Infected with Schistosoma japonicum

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Acta Biochimica et Biophysica Sinica 2006年第5期38卷 299-304页

作者： Xiao-Ping CAI Hui ZHANG Yong-Chen ZHANG Yong WANG Chuan SU min-jun ji Hai-Wei WU Xiang ZHU Zhao-Song ZHANG Guan-Ling WU Department of Pathogenic Biology,jiangsu Province Key Laboratoly of Modern Pathogenic Biology,Nanjing Medical University,Nanjing 210029,China Department of Pathogenic Biology Jiangsu Province Key Laboratory of Modern Pathogenic Biology Nanjing Medical UniversityNanjing 210029 China

CD4~+CD25~+T cells play a major role in modulating immune response,but few reports havebeen published about ***,we investigated the changes in CD4~+CD25~+T cell populations inspleens and mesenteric lymph nodes of mice infected with Schistosoma *** proportions ofCD4~+CD25~+T cells in total CD4~+T cells were analyzed by flow ***25 and Foxp3 expression wasmeasured by real-time quantitative polymerase chain *** suppressive activities of CD4~+CD25~+Tcells were detected by in vitro proliferation of *** showed that the percentage of CD4~+CD25~+T cells was the same as controls 3 weeks *** the acute stage of infection,the percentagedecreased ***,at the chronic stage of infection,it rebounded to normal levels or *** expression of the CD25 and Foxp3 showed gradual increase along with the infection *** experiment also showed the strong suppressive effect of CD4~+CD25~+T cells,isolated during the chronicstage,on proliferation of the CD25~-*** is the first time that the dynamics of CD4~+CD25~+T cellpopulations was demonstrated in mice infected with *** conclusion,our data indicated thatCD4~+CD25~+cells might be involved in the immune modulation during *** infection,which en-hances current knowledge of the mechanisms of the immuno-downregulation and re-infection inschistosomiasis.

关键词： Schistosoma japonicum CD4~+CD25~+ T cell chronic infection Foxp3

Effect of cholesterol metabolism on hepatolithiasis

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World Journal of Gastroenterology 2025年第1期31卷 157-162页

作者： Lin Zheng Zi-Yu Ye jun-ji Ma Department of Gastroenterology The Second Hospital of Hebei Medical UniversityHebei Key Laboratory of GastroenterologyHebei Institute of GastroenterologyHebei Clinical Research Center for Digestive DiseasesNo.215 Heping West RoadShijiazhuang 050000Hebei ProvinceChina

Surgical intervention is currently the primary treatment for hepatolithiasis;how-ever,some patients still experience residual stones and high recurrence rates after *** metabolism seems to play an important role in hepatoli-thiasis pathogenesis.A high cholesterol diet is one of the significant reasons for the increasing incidence of ***,regular diet and appropriate medical intervention are crucial measures to prevent hepatolithiasis and reduce recurrence rate after *** dietary cholesterol and drugs that increase cholesterol stone solubility are key therapeutic approaches in treating *** article discusses the cholesterol metabolic pathways related to the pathogenesis of hepatolithiasis,as well as food intake and targeted therapeutic drugs.

关键词： Hepatolithiasis Cholesterol metabolism High-fat diet 3-hydroxy-3-methylglutaryl-coenzyme A reductase Interlobular bile duct

Diagnostic pitfalls in malignant gliomas： the analysis of misdiagnosis and current recommendations

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Chinese Medical Journal 2012年第24期125卷 4520-4522页

作者： WANG Yu GUO Yi GAO jun DENG Kan LI Gui-lin FENG min WEI jun-ji XU Zhi-qin YAO Yong MA Wen-bin LI Yong-ning YANG Yi WANG Ren-zhi Department of Neurosurgery Peking Union Medical College Hospital Chinese Academy of Medical Sciences Beijing 100730 China

The malignant glioma is characterized by intrinsic aggressiveness and carries a dismal prognosis. Thecurrent standard treatment regimen for patients with malignant gliomas, specifically glioblastoma, is a combined therapy comprised of surgical resection followed by adjunctive radiation and chemotherapy. Yet, even with this new multimodality treatment, glioblastoma （GBM） recurs after a median time of 7 months following diagnosis, requiring a second-line treatment. But the clinical decisions are always difficult.

关键词： malignant gliomas diagnosis pseudoprogression radiation necrosis

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一种多群进化规划算法

数据采集与处理 2004年第3期19卷 258-263页

作者：张民王向军嵇斗海军工程大学电气工程系武汉430033

在分析了导致进化规划算法早熟原因的基础上 ,提出了一种改进的多群进化规划算法。在该算法中 ,进化在多个不同的子群间并行进行 ,通过使用不同的变异策略 ,实现种群在解空间具有尽可能分散探索能力的同时 ,在局部具有尽可能细致的搜索... 详细信息

在分析了导致进化规划算法早熟原因的基础上 ,提出了一种改进的多群进化规划算法。在该算法中 ,进化在多个不同的子群间并行进行 ,通过使用不同的变异策略 ,实现种群在解空间具有尽可能分散探索能力的同时 ,在局部具有尽可能细致的搜索能力。通过子群重组实现子群间的信息交换 ,基于典型算例的数字仿真证明 ,该算法具有更好的全局收敛性 ,更快的收敛速度和更强的鲁棒性。

关键词：进化规划 EP 搜索探索 MEP 数值仿真全局收敛性参数鲁棒性

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基于GPER2/MAPK/STAT1轴研究少腹逐瘀汤治疗子宫内膜异位症痛经分子机制

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中国中药杂志 2018年第16期43卷 3362-3367页

作者：崔宇红吴晶蔡玮杜娟曹军民吉秀家巩子汉甘肃中医药大学甘肃兰州730000 甘肃省人民医院甘肃兰州730000

该研究基于GPER2／MAPK／sTAT1轴研究少腹逐瘀汤治疗子宫内膜异位症痛经分子机制。采用异位移植造EMT模型；HE染色观察各组大鼠灌药治疗后宫腔组织病理变化；EEISA实验检测大鼠宫腔组织中TNF-α，IL一6的含量变化；qPCR检测宫腔组织中... 详细信息

该研究基于GPER2／MAPK／sTAT1轴研究少腹逐瘀汤治疗子宫内膜异位症痛经分子机制。采用异位移植造EMT模型；HE染色观察各组大鼠灌药治疗后宫腔组织病理变化；EEISA实验检测大鼠宫腔组织中TNF-α，IL一6的含量变化；qPCR检测宫腔组织中神经激肽受体（NK1）、GPER mRNA表达；Western blot法检测官腔组织中MAPK，STAT1蛋白含量。结果显示，在病理水平上，与模型组相比，少腹逐瘀汤可显著改善异位宫腔组织炎症情况，导致异位宫腔组织中浸润的炎症细胞减少，抑制血管生成，使宫腔组织壁变薄，阻止其生长，这种抑制表现出一定的浓度依赖性。在分子水平上，少腹逐瘀汤降低宫腔组织中炎症相关因子TNF-α，IL-6含量，减少NK1，GPER2 mRNA的相对表达量，并导致宫腔组织中MAPK和STAT1蛋白表达降低。综上可知少腹逐瘀汤可有效抑制异位宫腔组织中GPER2 mRNA以及MAPK，STAT1蛋白的表达，降低TNF-α，IL-6相关炎症因子的含量以及疼痛介质NK1 mRNA的含量从而减轻子宫内膜异位症患者炎症疼痛。

关键词：少腹逐瘀汤子宫内膜异位症痛经 GPER2/MAPK/sTAT1轴

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