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Amplification,cloning,sequencing and expression of the gene encoding the major surface antigen of Toxoplasma gondii isolated in China
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作者 陈晓光 刘国章 +2 位作者 徐帆 王章 江静波 《Journal of Medical Colleges of PLA(China)》 CAS 1994年第2期98-102,共5页
According to the published gene sequence of the major surface antigen (P30) of Toxoplasma gondii, a pair of primers were designed and synthesized. Using polymerase chain reaction (PCR), the coding sequences of P30 gen... According to the published gene sequence of the major surface antigen (P30) of Toxoplasma gondii, a pair of primers were designed and synthesized. Using polymerase chain reaction (PCR), the coding sequences of P30 gene were amplified from a Chinese strain of T. gondii, The amplified gene fragment and plasmid pB220 were digested with EcoRI and BamHI and then ligated. The inserted gene fragment was sequenced by the chain termination method, the reading reveals that nucleotide sequence determined was the same as the P30 sequecne of RH strain pubilished by Burg (1988), except that one base was changed. The recombinant plasmid containing P30 gene was transformed to E. coli DH5α.After temperature inducing culture, the total cellular proteins were analysed by SDS-PAGE and Western blot. The results show that the p30 gene cloned into the plasmid could express in E. coli, and the expression product had immunogenicity. 展开更多
关键词 TOXOPLASMA GONDII MAJOR surface antigen polymerase chain reaction gene CLONING sequence gene expression
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Construction of the Antisense Eukaryotic Vector for Proliferating Cell Nuclear Antigen Gene and Its Expression in Bladder Cancer EJ Cell Line
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作者 童强松 曾甫清 +2 位作者 齐义鹏 朱朝晖 鲁功成 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2002年第4期327-330,共4页
To explore a novel strategy for antisense gene therapy of cancer,the coding sequence of hum an proliferating cell nuclear antigen(PCNA) c DNA was reversely inserted into the eukaryotic vector p L XSN by molecular cl... To explore a novel strategy for antisense gene therapy of cancer,the coding sequence of hum an proliferating cell nuclear antigen(PCNA) c DNA was reversely inserted into the eukaryotic vector p L XSN by molecular cloning techniques and transferred into bladder cancer EJcells with li- posome. The PCNA expression in transferred cells was dynamically detected by immunofluo- rescence and RT- PCR techniques. Changes of proliferation activities of cancer cells were assayed by MTT colorim etric and cloning formation m ethods.In the experiment,the antisense eukaryotic vector was successfully constructed and nam ed as p L APSN.After transfection with it for1- 7 days,PCNA protein and m RNA levels in cancer cells were blocked by16 .74 % - 84 .2 1% (P< 0 .0 5 ) and2 3.2 7% - 86 .15 % (P<0 .0 5 ) respectively.The proliferation activities of transferred cells were inhibited by 2 7.91% - 6 2 .0 7% (P<0 .0 1) ,with cloning formation abilities being de- creased by 5 0 .81% (P<0 .0 1) . Itwas concluded that the in vitro proliferation activities of cancer cells could be effectively inhibited by blocking PCNA expression with antisense technique,which could serve as an ideal strategy for gene therapy of bladder cancer. 展开更多
关键词 proliferating cell nuclear antigen antisense RNA m olecular cloning gene expression
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Expression of core gene cDNA of Chinese hepatitis C virus in HepG2 cell line
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作者 姜荣龙 卢桥生 +3 位作者 侯金林 骆抗先 章廉 王燕军 《Journal of Medical Colleges of PLA(China)》 CAS 1997年第4期313-316,共4页
Objective: To clone core gene cDNA of Chinese hepatitis C virus (HCV) into eukaryotic expression vector pTM3 and to express HCV core antigen in HepG2 cells. Methods: Core gene cDNA of HCV was introduced into eukaryoti... Objective: To clone core gene cDNA of Chinese hepatitis C virus (HCV) into eukaryotic expression vector pTM3 and to express HCV core antigen in HepG2 cells. Methods: Core gene cDNA of HCV was introduced into eukaryotic expression vector pTM3. Using vaccinia virus/bacteriophage T7 hybrid expression system, HepG2cells were trans feeted with the recombinant plasmid pTM3-Q534 by lipofectin. Results: From the transfected bacteria Top10F, 2pTM3-Q534 clones containing the recombinant plasmid were identified from randomly selected 10ampicillin-resistant colonies. By indirect immunofluorescence technique HCV core protein was identified in HepG2cells trans feeted with the recombinant plasmid. Conclusion: The construction of a recombinant plasmid and the expression of core gene cDNA of HCV in HepG2 were successful. 展开更多
关键词 HEPATITIS C VIRUS (HCV)/core gene CDNA cosmid vector CLONAL expression
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Prokaryotic Expression and Antigenic Analysis of Wbkc Gene from Brucella abortus
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作者 Xiangyang LI Xueli WANG +4 位作者 Kai LIU Xiaowei HUO Yinghong WU Xianhua ZHANG Jiabao ZHANG 《Agricultural Biotechnology》 CAS 2014年第2期51-53,共3页
[ Objective ] This study aimed to clone and express formyltransferase (Wbkc) gene from Brucella abortus in E. coli, purify the expressed protein and analyze its immunogenicity. [Method] A gene encoding 27 -35 ku for... [ Objective ] This study aimed to clone and express formyltransferase (Wbkc) gene from Brucella abortus in E. coli, purify the expressed protein and analyze its immunogenicity. [Method] A gene encoding 27 -35 ku formyltransferase (Wbkc) was amplified from the genomic DNA of BruceUa abortus by PCR. The amplified fragments were digested with BamH I and Sal I, and inserted into pET28a vector. The constructed recombinant plasmid pET 28a-Wbkc was trans- formed into E. coli BL21 and was induced to express the fusion protein. Subsequently, the protein was purified by histidine-binding resin column chromatography, and the immunogenicity was detected by Western blot assay. The recombinant plasmid was identified by colony PCR, double digestion and sequencing analysis. [ Result] Wbkc was successfully cloned and expressed in E. coli. A specific protein band of 29 ku was detected by SDS-PAGE. Western blot showed specific im- munoreactivity of the purified fusion protein. [ Conclusion] This study provided a solid foundation for further investigating diagnostic proteins for brucellosis and developing Brucella gene-deletion vaccines. 展开更多
关键词 Brucella abortus Formyltransferase (Wbkc) gene Prokaryotic expression antigenICITY
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Novel DNA vaccine based on hepatitis B virus core gene induces specific immune responses in Balb/c mice 被引量:7
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作者 Yi-Ping Xing Zu-Hu Huang +4 位作者 Shi-Xia Wang Jie Cai Jun Li Te-Hui W Chou Shan Lu 《World Journal of Gastroenterology》 SCIE CAS CSCD 2005年第29期4583-4586,共4页
AIM: To investigate the immunogenidty of a novel DNA vacoine, pSW3891/HBc, based on HBV core gene in Balb/c mice. METHODS: A novel DNA vaccine, pSW3891/HBc, encoding HBV core gene was constructed using a vector plas... AIM: To investigate the immunogenidty of a novel DNA vacoine, pSW3891/HBc, based on HBV core gene in Balb/c mice. METHODS: A novel DNA vaccine, pSW3891/HBc, encoding HBV core gene was constructed using a vector plasmid pSW3891. Balb/c mice were immunized with either pSW3891/HBc or empty vector DNA via gene gun. IgG anti-HBc responses in mouse sera were demonstrated by ELISA. Specific cytotoxicity of cytotoxic T lymphocytes (CTLs) of mice was quantitatively measured by lactate dehydrogenase release assay. RESULTS: HBcAg was expressed effectively in 293T cell line transiently transfected with pSW3891/HBc. Strong IgG anti-HBc responses were elicited in mice immunized with pSW3891/HBc. The end-point titers of anti-HBc reached the highest 1:97 200, 4 wk after the third immunization. The specific CTL killing with the highest specific lysis reached 73.25% at effector:target ratio of 20:1 in mice that received pSW3891/HBc DNA vaccine. CONCLUSION: pSW3891/HBc vaccination elicits specific anti-HBc response and induces HBc-specific CTL response in immunized Balb/c mice. 展开更多
关键词 DNA vaccine Hepatitis B virus core antigen IMMUNOGENICITY gene gun CTL HBV
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Inhibition of hepatitis B virus surface antigen expression by small hairpin RNA in vitro 被引量:8
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作者 Zheng-GangYang ZhiChen QinNi NingXu Jun-BinShao Hang-PingYao 《World Journal of Gastroenterology》 SCIE CAS CSCD 2005年第4期498-502,共5页
AIM: To explore the anti-hepatitis B virus effect of RNA interference (RNAi) using small hairpin RNA (shRNA)expression vector.METHODS: Hepatitis B virus surface antigen green fluorescent protein (HBs-GFP) fusion vecto... AIM: To explore the anti-hepatitis B virus effect of RNA interference (RNAi) using small hairpin RNA (shRNA)expression vector.METHODS: Hepatitis B virus surface antigen green fluorescent protein (HBs-GFP) fusion vector and shRNA expression vectors were constructed and cotransfected transiently into HepG2 cells. mRNAs extracted from HepG2 cells were detected by real-time PCR. Fluorescence of HBs-GFP protein was detected by fluorescence-activated cell sorting (FACS). The effective shRNA expression vector was transfected into HepG2.2.15 cells. HBsAg and HBeAg in HepG2.2.15 cells were analyzed by radioimmunoassay (RIA) method.RESULTS: FACS revealed that shRNA targeting at HBsAg reduced the GFP signal by 56% compared to the control.Real-time PCR showed that HBs-GFP mRNA extracted from HepG2 cells cotransfected with pAVU6+27 and HBs-GFP expression plasmids decreased by 90% compared to the empty vector control. The expressions of HBsAg and HBeAg were also inhibited by 43% and 64%, respectively.CONCLUSION: RNAi using shRNA expression vector can inhibit the expression of HBsAg, providing a fresh approach to screening the efficient small interfering RNAs (siRNAs). 展开更多
关键词 Hepatitis B Surface antigens Small hairpin RNA RNA interference gene expression
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Transretinoic acid inhibits rats gastric epithelial dysplasia induced by N-methyi-N-nitro-N-nitrosoguanidine:influences on cell apoptosis and expression of its regulatory genes 被引量:8
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作者 Ru Tao Cui Gan Cai +3 位作者 Zhao Bao Yin Yong Cheng Qiu Hong Yang Tao Tian 《World Journal of Gastroenterology》 SCIE CAS CSCD 2001年第3期394-398,共5页
INTRODUCTIONGastric epithelial dysplasia (GED) hypothetically is a straight-forward concept: dysplastic epithelium replacing the normal gastric epithelium of the stomach [1].In the stomach ,like any other segment of t... INTRODUCTIONGastric epithelial dysplasia (GED) hypothetically is a straight-forward concept: dysplastic epithelium replacing the normal gastric epithelium of the stomach [1].In the stomach ,like any other segment of the gut ,it is defined as an unequivocal non-invasive epithelial change[2,3].The observation of gastric dysplasia as a cancerous lesion was recognized over a century ago ,but it is only after the advent of gastroscopy that its clinical significance has been stressed[4-7]. 展开更多
关键词 Animals antigens CD95 Antineoplastic Agents Apoptosis Caspase 1 Cyclin D1 Gastric Mucosa gene expression Immunohistochemistry Male Membrane Glycoproteins METHYLNITRONITROSOGUANIDINE RNA Messenger RATS Rats Wistar Research Support Non-U.S. Gov't Stomach Diseases TRETINOIN
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PRAME Gene Expression in Acute Leukemia and Its Clinical Significance 被引量:1
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作者 Kai Ding Xiao-ming Wang +3 位作者 Rong Fu Er-bao Ruan Hui Liu Zong-hong Shao 《Clinical oncology and cancer researeh》 CAS CSCD 2012年第1期73-76,共4页
Objective To investigate the expression of the preferentially expressed antigen of melanoma (PRAME) gene in acute leukemia and its clinical significance. Methods The level of expressed PRAME mRNA in bone marrow mono... Objective To investigate the expression of the preferentially expressed antigen of melanoma (PRAME) gene in acute leukemia and its clinical significance. Methods The level of expressed PRAME mRNA in bone marrow mononuclear cells from 34 patients with acute leukemia (AL) and in 12 bone marrow samples from healthy volunteers was measured via RT-PCR. Correlation analyses between PRAME gene expression and the clinical characteristics (gender, age, white blood count, immunophenotype of leukemia, percentage of blast cells, and karyotype) of the patients were performed. Results The PRAME gene was expressed in 38.2% of all 34 patients, in 40.7% of the patients with acute myelogenous leukemia (AML, n=27), and in 28.6% of the patients with acute lymphoblastic leukemia (ALL, n=7), but was not expressed in the healthy volunteers. The difference in the expression levels between AML and ALL patients was statistically significant. The rate of gene expression was 80% in M~, 33.3% in M2, and 28.6% in M~. Gene expression was also found to be correlated with CDl5 and CD33 expression and abnormal karyotype, but not with age, gender; white blood count or percentage of blast cells. Conclusions The PRAME gene is highly expressed in acute leukemia and could be a useful marker to monitor minimal residual disease. This gene is also a candidate target for the immunotherapy of acute leukemia. 展开更多
关键词 preferentially expressed antigen of melanoma gene acute leukemia minimal residual disease IMMUNOTHERAPY
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Expression of Core Domain of Porcine Zona Pellucida 3β in E.coli
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作者 Qiu-ling XIE Xiao-jia CHEN +5 位作者 Wei-jie ZHU Ling ZHANG Wan-xiang XU An HONG Jing LI Si-hong GAO 《Journal of Reproduction and Contraception》 CAS 2005年第2期67-72,共6页
To obtain the recombinant core domain of porcine zone pellucida 3β (cZP3β) for the further research on its functions Methods The nucleotide sequence region from 44 to 306 codons of pZP3β entire eDNA was obtained ... To obtain the recombinant core domain of porcine zone pellucida 3β (cZP3β) for the further research on its functions Methods The nucleotide sequence region from 44 to 306 codons of pZP3β entire eDNA was obtained by PCR and then was cloned into pET-3c vector. After being identified, recon was transformed into E.coli BL21 (DE3) pLysS and then induced by IPTG. Results The recombinant cZP3β was expressed in E. coli up to 15% of total cellular proteins, and was made sure by Western blot analysis. Conclusion The research on expression of core domain of pZP3β could benefit to further investigation of its immunogenicity and the development of antigen preparation. 展开更多
关键词 porcine zona pellucida core domain E. coli gene expression porcine zona pellucida core domain E. coli gene expression
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Construction and Expression of Prokaryotic Expression Vector of MPT-64 Gene
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作者 Long PENG Linbo ZHANG 《Agricultural Biotechnology》 CAS 2014年第3期11-13,17,共4页
[Objective]Protective antigen gene MPT-64 was cloned from genomic DNA of Mycobacterium tuberculosis and transferred into prokaryotic competent cells for expression to obtain MPT-64 fusion protein.[Method]Based on the ... [Objective]Protective antigen gene MPT-64 was cloned from genomic DNA of Mycobacterium tuberculosis and transferred into prokaryotic competent cells for expression to obtain MPT-64 fusion protein.[Method]Based on the GenBank,primers were designed for amplification of MPT-64 gene,and the recombinant plasmid pET-32a-MPT-64 was constructed.The recombinant plasmid was expressed in prokaryotic expression vector to obtain fusion protein.[Result]Protective antigen gene MPT-64 was successfully cloned.The recombinant plasmid pET-32a-MPT-64 was obtained.MPT-64 fusion protein was successfully expressed.[Conclusion]This study laid solid foundation for the prevention,diagnosis,treatment of tuberculosis and the development of tuberculosis vaccines. 展开更多
关键词 Mycobacterium tuberculosis Protective antigen genes Secreted protein MPT64 Prokaryotic expression Fusion protein
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Cloning of human melanoma antigen MAGE-A9 and its expression in hepatocellular carcinomas
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作者 Lu Xu Jin Zhu +2 位作者 Zhenning Qiu Yuhua Li Zhenqin Feng 《Journal of Nanjing Medical University》 2007年第5期287-292,共6页
Objective:To express the melanoma associated gene MAGE-A9 recombinant protein, obtain the anti-MAGE-A9 monoclonal antibody and to examine the expression of MAGE-A9 in hapatocellular carcinoma specimens. Methods:MAGE... Objective:To express the melanoma associated gene MAGE-A9 recombinant protein, obtain the anti-MAGE-A9 monoclonal antibody and to examine the expression of MAGE-A9 in hapatocellular carcinoma specimens. Methods:MAGE-A9 cDNA was cloned from human hepatocellular carcinoma tissue by using RT-PCR, and then subcloned into the plasmid pMD18-T. After sequencing, the MAGE-A9 was cloned into the prokaryotic expression vector pBAD/gⅢ to construct the recombinant expression vector pBAD/gⅢ - MAGE-A9, and was transformed into E. coli TOP10. The recombinant MAGE-A9 protein was expressed under induction of L-Arabinose, and was purified through Hitrap column. The anti-MAGE-A9 monoclonal antibody was generated. The expression of MAGE-A9 in hepatocellular carcinoma specimens was examined through ABC assay. Results:The cDNA sequence of the cloned MAGE-A9 gene was consistent with the reported sequence. By affinity column and SDS-PAGE, the purified MAGE-A9 fusion protein displayed a band of Mr 35,000, and subsequently the anti-MAGE-A9 monoclonal antibody was obtained. We found that MAGE-A9 expressed in the cytoplast of positive cells and MAGE-A9 antigen was detected in 8 cases out of 39 (21%) hepatocellular carcinoma specimens. Conclusion:MAGE-A9 antigen was expressed in a fair proportion of hepatocellular carcinoma specimens, these patients might be suitable candidates for immune involving antigen, encoded by the MAGE-A9 gene. 展开更多
关键词 melanoma antigen MAGE-A9 RT-PCR gene expression IMMUNOHISTOCHEMISTRY
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STRUCTURE AND EXPRESSION OF EPSTEIN-BARR VIRUS MEMBRANE ANTIGEN IN RECOMBINANT VACCINIA VIRUS
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作者 谷淑燕 江民康 +4 位作者 赵文平 曾毅 侯云德 朱既明 Hans Wolf 《Chinese Journal of Cancer Research》 SCIE CAS CSCD 1989年第1期44-49,共6页
The Epstein-Barr virus membrane antigen was constructed and inserted into vaccinia virus, Tian-tan strain in order to study the effect of this virus on EB infection and tumorogenesis. The EBV-derived membrane antigen ... The Epstein-Barr virus membrane antigen was constructed and inserted into vaccinia virus, Tian-tan strain in order to study the effect of this virus on EB infection and tumorogenesis. The EBV-derived membrane antigen was expressed under the control of a 7.5 K promoter of vaccinia virus. The antibody against the membrane antigen of EB virus was produced on rabbits vaccinated with recombinant vaccinia virus. 展开更多
关键词 EBV MA STRUCTURE AND expression OF EPSTEIN-BARR VIRUS MEMBRANE antigen IN RECOMBINANT VACCINIA VIRUS gene
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骨关节炎核心基因的生物信息学鉴定
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作者 朱雪坤 刘恒 +5 位作者 冯晖 高云龙 文磊 蔡筱松 赵奔 仲敏 《中国组织工程研究》 CAS 北大核心 2025年第3期637-644,共8页
背景:目前骨关节炎成为影响老年人生活质量的主要疾病,治疗效果欠佳,临床治疗措施主要集中在阻止疾病的进程,同时骨关节炎的发病机制尚不完全清楚。为了探索骨关节炎的主要发病机制和基因编码调控的相关机制,进行生物信息学分析。目的:... 背景:目前骨关节炎成为影响老年人生活质量的主要疾病,治疗效果欠佳,临床治疗措施主要集中在阻止疾病的进程,同时骨关节炎的发病机制尚不完全清楚。为了探索骨关节炎的主要发病机制和基因编码调控的相关机制,进行生物信息学分析。目的:通过基因表达谱筛选出在骨关节炎中起主要作用的核心差异基因。方法:从基因表达综合数据库(GEO)下载数据集:GSE114007、GSE117999和GSE129147,利用R软件筛选出GSE114007和GSE117999数据合集的差异基因,将差异基因进行加权基因共表达网络分析,选择和骨关节炎最相关的模块基因并进行蛋白互作分析,利用cytocape软件筛选出候选核心基因,随后将候选核心基因进行最小绝对收缩和选择算子回归(LASSO回归)和COX分析,鉴定出在骨关节炎中起关键作用的核心基因,利用外部数据集GSE129147验证核心基因的准确性。结果与结论:①共鉴定出477个差异基因,加权基因共表达网络分析获得265个和骨关节炎相关的差异基因,共鉴定8个候选核心基因,LASSO回归分析最终获得了一个具有核心价值的差异基因ASPM并进行了外部验证;②提示通过生物信息学筛选出基因ASPM表达异常在骨关节炎中起到关键核心作用。 展开更多
关键词 骨关节炎 差异表达基因 核心差异基因 生物信息学 LASSO回归分析
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丙型肝炎病毒Core与NS3基因的不同融合方式及其表达 被引量:1
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作者 李暐东 梁布锋 祁自柏 《中国病毒学》 CSCD 2000年第2期204-207,共4页
The gene encoding the core and NS3 proteins of hepatitis C virus was amplified by PCR, respectirely. Two genes were fused and formed the recombinant plamid pHCV CN (Core+NS3) and pHCV NC (NS3+Core). The fused plasmid ... The gene encoding the core and NS3 proteins of hepatitis C virus was amplified by PCR, respectirely. Two genes were fused and formed the recombinant plamid pHCV CN (Core+NS3) and pHCV NC (NS3+Core). The fused plasmid were expressed in E.coli 06. The pHC CN plasmid expressed fussion protein was shown by a major band with a expected molecular weight about 68 kD on SDS PAGE. However, the pHCV NC plasmid expressed fussion protein was 66 kD in molecular weight. The results indicate that the pHCV NC fussion protein differs from the pHCV CN fussion protein in mulecular weight. It suggests that the fusion way is important for the structure of recombinant proteins. 展开更多
关键词 丙型肝炎病毒 core基因 NS3基因 基因融合 表达
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Expression of lipopolysaccharide binding protein and its receptor CD14 in experimental alcoholic liver disease 被引量:14
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作者 Guo-Qing Zuo~1 Jian-Ping Gong~2 Chang-An Liu~2 Shen-Wei Li~2 Xin-Chuan Wu~2 Kang Yang~2 Yue Li~2 1 Department of Digestive Disease2 Department of General Surgery,Second College of Clinical Medicine &the Second Affiliated Hospital,Chongqing University of Medical Sciences,Chongqing 400010,China 《World Journal of Gastroenterology》 SCIE CAS CSCD 2001年第6期836-840,共5页
AIM: To evaluate the relationship between the expression of lipopolysaccharides (LPS) binding protein (LBP) and CD14 mRNA and the severity of liver injury in alcohol-fed rats. METHODS: Twenty Wistar rats were divided ... AIM: To evaluate the relationship between the expression of lipopolysaccharides (LPS) binding protein (LBP) and CD14 mRNA and the severity of liver injury in alcohol-fed rats. METHODS: Twenty Wistar rats were divided into two groups:ethanol-fed group (group E) and control group (group C). Group E was fed with ethanol(5-12 g x kg(-1) x d(-1)) and group C received dextrose instead of ethanol. Rats of the two groups were sacrificed at 4 weeks and 8 weeks. Levels of endotoxin and alanine transaminase (ALT) in blood were measured, and liver pathology was observed under light and electronic microscopy. Expressions of LBP and CD14 mRNA in liver tissues were determined by RT-PCR analysis. RESULTS: Plasma endotoxin levels were increased more significantly in group E(129+/-21) ng x L(-1) and (187+/-35) ng x L(-1) at 4 and 8 wk than in control rats(48+/-9) ng x L(-1) and (53+/-11) ng x L(-1), respectively (P【0.05). Mean values of plasma ALT levels were (1867+/-250) nkat x L(-1) and (2450+/-367) nkat x L(-1) in Group E. The values were increased more dramatically in ethanol-fed rats than in Group C after 4 and 8 weeks. In liver section from ethanol-fed rats, there were marked pathological changes (steatosis, cell infiltration and necrosis). In ethanol-fed rats, ethanol administration led to a significant increase in LBP and CD14 mRNA levels compared with the control group (P【0.05). CONCLUSION: Ethanol administration led to a significant increase in endotoxin levels in serum and LBP and CD14 mRNA expressions in liver tissues. The increase of LBP and CD14 mRNA expression might wake the liver more sensitive to endotoxin and liver injury. 展开更多
关键词 Acute-Phase Proteins gene expression Membrane Glycoproteins Animals antigens CD14 Carrier Proteins Female Liver Liver Diseases Alcoholic RNA Messenger RATS Rats Wistar Research Support Non-U.S. Gov't Severity of Illness Index
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Relationship between Fas/ FasL expression and apoptosis of colon adenocarcinoma cell lines 被引量:15
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作者 Zhi Hai Peng Tong Hai Xing +1 位作者 Guo Qiang Qiu Hua Mei Tang Shanghai No. 1 People’s Hospital, Shanghai 200080, China 《World Journal of Gastroenterology》 SCIE CAS CSCD 2001年第1期88-92,共5页
INTRODUCTIONFas/ FasL system has been identified as a keymediator of apoptosis in tumor cells[1-4]. Theoccurrence and development of neoplasm are closelyrelated to apoptosis[5-7] Most chemotherapeuticdrugs kill cancer... INTRODUCTIONFas/ FasL system has been identified as a keymediator of apoptosis in tumor cells[1-4]. Theoccurrence and development of neoplasm are closelyrelated to apoptosis[5-7] Most chemotherapeuticdrugs kill cancer cells mainly by inducingapoptosis[8-14].' 展开更多
关键词 Adenocarcinoma Colonic Neoplasms Antibiotics Antineoplastic antigens CD95 Antimetabolites Antineoplastic Antineoplastic Agents APOPTOSIS Cisplatin EPIRUBICIN Flow Cytometry Fluorouracil gene expression Regulation Neoplastic Humans Membrane Glycoproteins Mitomycins Research Support Non-U.S. Gov't Tumor Cells Cultured
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Helicobacter pylori HopE and HopV porins present scarce expression among clinical isolates
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作者 Maritza Lienlaf Juan Pablo Morales +6 位作者 María Inés Díaz Rodrigo Díaz Elsa Bruce Freddy Siegel Gloria León Paul R Harris Alejandro Venegas 《World Journal of Gastroenterology》 SCIE CAS CSCD 2010年第3期320-329,共10页
AIM:To evaluate how widely Helicobacter pylori (H. pylori ) HopE and HopV porins are expressed among Chilean isolates and how seroprevalent they are among infected patients in Chile.METHODS: H. pylori hopE and hopV ge... AIM:To evaluate how widely Helicobacter pylori (H. pylori ) HopE and HopV porins are expressed among Chilean isolates and how seroprevalent they are among infected patients in Chile.METHODS: H. pylori hopE and hopV genes derived from strain CHCTX-1 were cloned by polymerase chain reaction (PCR), sequenced and expressed in Escherichia coli AD494 (DE3). Gel-purified porins were used to prepare polyclonal antibodies. The presence of both genes was tested by PCR in a collection of H. pylori clinical isolates and their expression was detected in lysates by immunoblotting. Immune responses against HopE, HopV and other H. pylori antigens in sera from infected and non-infected patients were tested by Western blotting using these sera as f irst antibody on recombinant H. pylori antigens.RESULTS: PCR and Western blotting assays revealed that 60 and 82 out of 130 Chilean isolates carried hopE and hopV genes, respectively, but only 16 and 9, respectively, expressed these porins. IgG serum immunoreactivity evaluation of 69 H. pylori-infected patients revealed that HopE and HopV were infrequently recognized (8.7% and 10.1% respectively) compared to H. pylori VacA (68.1%) and CagA (59.5%) antigens. Similar values were detected for IgA serum immunoreactivity against HopE (11.6%) and HopV (10.5%) although lower values for VacA (42%) and CagA (17.4%) were obtained when compared to the IgG response.CONCLUSION: A scarce expression of HopE and HopV among Chilean isolates was found, in agreement with the infrequent seroconversion against these antigens when tested in infected Chilean patients. 展开更多
关键词 Helicobacter pylori gene expression HopE and HopV porins antigenS Immune response
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Differential expression and significance of 5-hydroxymethylcytosine modification in hepatitis B virus carriers and patients with liver cirrhosis and liver cancer
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作者 Yue-Cui Li Wei-Yue Hu +4 位作者 Cheng-Hang Li Li-Li Zhang Xiang-Wei Xu Jin Li Hong-Xia Luo 《World Journal of Gastrointestinal Surgery》 SCIE 2023年第3期346-361,共16页
BACKGROUND The relationship between hepatitis B surface antigen(HBsAg)-positive carrier status and liver cancer has been extensively studied.However,the epigenetic changes that occur during progression from HBsAg-posi... BACKGROUND The relationship between hepatitis B surface antigen(HBsAg)-positive carrier status and liver cancer has been extensively studied.However,the epigenetic changes that occur during progression from HBsAg-positive carrier status or cirrhosis to liver cancer are unknown.The epigenetic modification of DNA hydroxymethylation is critical in tumor development.Further,5-hydroxymethylcytosine(5hmC)is an important base for DNA demethylation and epigenetic regulation.It is also involved in the assembly of chromosomes and the regulation of gene expression.However,the mechanism of action of 5hmC in HBsAgpositive carriers or patients with cirrhosis who develop liver cancer has not been fully elucidated.AIM To investigate the possible epigenetic mechanism of HBsAg-positive carriers and hepatocellular carcinoma(HCC)progression from cirrhosis.METHODS Forty HBsAg-positive carriers,forty patients with liver cirrhosis,and forty patients with liver cancer admitted to the First People's Hospital of Yongkang between March 2020 and November 2021 were selected as participants.Free DNA was extracted using a cf-DNA kit.cfDNA was extracted by 5hmC DNA sequencing for principal component analysis,the expression profiles of the three groups of samples were detected,and the differentially expressed genes(DEGs)modified by hydroxymethylation were screened.Bioinformatic analysis was used to enrich DEGs,such as in biological pathways.RESULTS A total of 16455 hydroxymethylated genes were identified.Sequencing results showed that 32 genes had significant 5hmC modification differences between HBsAg carriers and liver cancer patients,of which 30 were upregulated and 2 downregulated in patients with HCC compared with HBsAg-positive carriers.Significant 5hmC modification differences between liver cirrhosis and liver cancer patients were identified in 20 genes,of which 17 were upregulated and 3 were downregulated in patients with HCC compared with those with cirrhosis.These genes may have potential loci that are undiscovered or unelucidated,which contribute to the development and progression of liver cancer.Analysis of gene ontology enrichment and Kyoto Encyclopedia of Genes and Genomes showed that the major signaling pathways involved in the differential genes were biliary secretion and insulin secretion.The analysis of protein interactions showed that the important genes in the protein-protein interaction network were phosphoenolpyruvate carboxykinase and solute carrier family 2.CONCLUSION The occurrence and development of liver cancer involves multiple genes and pathways,which may be potential targets for preventing hepatitis B carriers from developing liver cancer. 展开更多
关键词 Hepatitis B surface antigen 5-hydroxymethylcytosine Hepatocellular carcinoma Liver cancer DNA sequencing Differentially expressed genes
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IL-7的诱导表达增强靶向GPC3 CAR-T细胞的增殖及体外抗肿瘤活性
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作者 龚福生 陈珊珊 +1 位作者 郑秋红 刘沁颖 《中国肿瘤生物治疗杂志》 CAS CSCD 北大核心 2024年第10期951-956,共6页
目的:探讨IL-7的诱导表达对靶向磷脂酰肌醇蛋白聚糖3(GPC3)嵌合抗原受体基因修饰T淋巴细胞(CAR-T细胞)的增殖和体外抗肿瘤活性的影响。方法:通过无缝克隆将GPC3 CAR序列片段插入GV400载体的Bam HⅠ/Eco RⅠ位置,构建第二代CAR慢病毒载体... 目的:探讨IL-7的诱导表达对靶向磷脂酰肌醇蛋白聚糖3(GPC3)嵌合抗原受体基因修饰T淋巴细胞(CAR-T细胞)的增殖和体外抗肿瘤活性的影响。方法:通过无缝克隆将GPC3 CAR序列片段插入GV400载体的Bam HⅠ/Eco RⅠ位置,构建第二代CAR慢病毒载体GPC3-BBZ及GPC3-BBZ-NFAT-IL-7,以293T细胞包装相应的慢病毒载体后,感染人T细胞制备CAR-T细胞。实验分为未转导T细胞(NT)组、GPC3-BBZ CAR-T细胞组、GPC3-BBZ-NFAT-IL-7 CAR-T细胞组。采用流式细胞术检测各组CAR-T细胞中CAR的表达水平,qPCR法检测经GPC3蛋白激活的CAR-T细胞中IL-7 m RNA的表达水平,细胞计数法检测CAR-T细胞在GPC3抗原刺激下的增殖能力,ELISA检测CAR-T细胞在受到肿瘤细胞刺激后IL-7、IFN-γ和TNF-α的分泌水平。应用实时细胞分析(RTCA)技术检测CAR-T细胞对人肝癌Huh-7细胞的杀伤作用。结果:成功构建慢病毒载体GPC3-BBZ和GPC3-BBZ-NFAT-IL-7,制备出靶向GPC3的CAR-T细胞。经GPC3抗原激活后,GPC3-BBZ-NFAT-IL-7 CAR-T细胞可有效表达IL-7 mRNA(P<0.01),其表现出更强的增殖能力(P<0.05)。与GPC3-BBZ CAR-T细胞相比,GPC3-BBZ-NFAT-IL-7 CAR-T细胞与GPC3阳性靶细胞Huh-7细胞共培养后,分泌更高水平的IL-7、IFN-γ和TNF-α(P<0.01或P<0.001)。RTCA结果显示,GPC3-BBZ-NFAT-IL-7 CAR-T细胞对GPC3阳性Huh-7细胞的杀伤活性显著高于GPC3-BBZ CAR-T细胞(P<0.05)。结论:成功制备可诱导表达IL-7的靶向GPC3的CAR-T细胞,IL-7的诱导表达增强靶向GPC3 CAR-T细胞的免疫活性,在体外展现出较强的肿瘤细胞杀伤能力。 展开更多
关键词 肝细胞癌 磷脂酰肌醇蛋白聚糖3 CAR-T细胞 IL-7 诱导表达
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m^(6)A相关基因在激素性股骨头坏死中的生物信息学分析 被引量:2
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作者 令狐熙涛 桂佳琦 +2 位作者 梁卓智 瓦庆德 黄帅 《中国组织工程研究》 CAS 北大核心 2024年第36期5811-5816,共6页
背景:m^(6)A修饰与股骨头坏死的发生发展相关,但在激素性股骨头坏死中的作用尚不清楚。目的:基于GEO数据库,采用生物信息学方法分析激素性股骨头坏死中表达差异的m^(6)A基因及互作miRNAs,探寻其潜在发病机制。方法:在GEO数据库中检索并... 背景:m^(6)A修饰与股骨头坏死的发生发展相关,但在激素性股骨头坏死中的作用尚不清楚。目的:基于GEO数据库,采用生物信息学方法分析激素性股骨头坏死中表达差异的m^(6)A基因及互作miRNAs,探寻其潜在发病机制。方法:在GEO数据库中检索并下载与激素性股骨头坏死相关的mRNA表达谱数据集(GSE123568),通过R软件对数据集进行差异基因筛选及GO功能、KEGG通路富集分析。识别差异基因中的m^(6)A差异表达基因(m^(6)A-DEGs)并对其进行GO功能与KEGG通路富集分析,比较m^(6)A-DEGs的表达量并分析它们之间的相关性。最后通过Cytoscape构建m^(6)A-DEGs的PPI互作网络及筛选核心基因。使用TargetScan,miRTarBase和miRBD数据库预测m^(6)A-DEGs相关的潜在miRNAs,同时使用ChIPBase及hTFtarget数据库预测7个核心基因潜在转录因子,然后分别构建m^(6)A-miRNA与转录因子m^(6)A调控网络。最后使用数据集GSE74089验证7个核心m^(6)A-DEGs的表达水平。结果与结论:①从数据集中共筛选出2460个差异表达的基因,其中1455个上调,1005个下调。②从数据集中筛选出了14个m^(6)A-DEGs,包括3个下调和11个上调基因,m^(6)A-DEGs在激素性股骨头坏死中的表达具有显著差异(P<0.05),Spearman分析表明它们之间具有一定相关性。③m^(6)A-DEGs的GO和KEGG富集分析主要集中在骨髓细胞分化与发育、免疫受体与细胞因子受体活性、破骨细胞分化、AMPK与白细胞介素17信号通路。④m^(6)A-DEGs前7个核心基因包括YTHDF3,YTHDF1,YTHDF2,ALKBH5,METTL3,HNRNPA2B1及HNRNPC,它们在miRTarBase,miRDB和TargetScan数据库中共有44个miRNA重叠,在ChIPBase及hTFtarget数据库中共有79个重叠转录因子。⑤在GSE74089数据集中有6个核心m^(6)A-DEGs的表达水平与GSE123568数据集一致。⑥结果证实,根据生物信息学方法筛选的7个m^(6)A-DEGs可能通过调控多个miRNA、转录因子和AMPK及白细胞介素17信号通路表达,进而影响激素性股骨头坏死中骨髓细胞分化发育与破骨细胞分化,为进一步深入研究激素性股骨头坏死的发病机制和靶向治疗提供了数据支持和研究方向。 展开更多
关键词 激素性股骨头坏死 m^(6)A甲基化 微小RNA 转录因子 生物信息学 差异基因 基因调控网络 核心基因
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