检索结果-南通市图书馆

immortalized Rat Astrocyte Strain Genetically Modified by Rat Preprogalanin Gene

Journal of Huazhong University of Science and Technology(Medical Sciences) 2005年第2期25卷 144-146,197页

作者：安珂田玉科杨辉高峰王鹏 Department of Anesthesiology Tongji Hospital Tongji Medical College Huazhong Universityof Science and Technology Wuhan 430030 China

Summary: To construct an immortalized rat astrocyte strain genetically modified by rat preprogalanin gene (IAST/GAL) and detect its galanin (GAL) expression and secretion, a cDNA fragment of rat GAL in plasmid of pBS KS(+)-GAL was inserted into eukaryotic expression vector pcDNA3.1(+) by DNA recombinant technology, then the restriction enzyme digestion and DNA sequencing were carried out to evaluate the recombinant. The pcDNA3.1(+)-GAL and pcDNA3.1(+) construct were transfected into immortalized rat astrocyte strain (IAST) by lipofectamine and the population of cells which stably integrated the construct was selected with 600 μg/mL G418. Individual clones were screened and expanded into clonal cell strains. Detection of Neo gene was used to validate the success of the transfection. Immunocytochemical staining, RT-PCR and radioimmunoassay were used to detect the expression and secretion level of GAL. The recombinant had been successfully constructed by restriction enzyme digestion and DNA sequencing. Detection of Neo gene showed that the pcDNA3.1(+)-GAL and pcDNA3.1(+) have been successfully transfected into IAST. After selection by using G418, IAST/GAL and IAST/Neo cell strains were obtained. IAST/GAL, IAST/Neo and IAST were immunostained positively for GAL, but the GAL average optical density of IAST/GAL was significantly higher than that of IAST/Neo and IAST (P0.05). It was concluded that IAST/GAL strain was constructed successfully and it might provide a basis for the further study of pain therapy.

关键词： preprogalanin astrocyte immortalized cell strain biologic minipumps

来源：

维普期刊数据库

同方期刊数据库评论

在线全文

学校读者我要写书评

暂无评论

Expression of hepatitis B virus 1.3-fold genome plasmid in an SV40 T-antigen-immortalized mouse hepatic cell line

引用

World Journal of Gastroenterology 2013年第44期19卷 8020-8027页

作者： Xiu-Guang Song Peng-Fei Bian Shu-Li Yu Xiu-Hua Zhao Wei Xu Xue-Hui Bu Xia Li Li-Xian Ma Department of Infectious Diseases Qilu Hospital Shandong University Jinan Infectious Disease Hospital Shandong University Laboratory for Tumor Immunity and Traditional Chinese Drug Immunity Institute of Basic Medicine Shandong Academy of Medical Science School of Medicine Shandong University

AIM:To investigate the expression of the hepatitis B virus(HBV)1.3-fold genome plasmid(pHBV1.3)in an immortalized mouse hepatic cell line induced by SV40T-antigen(SV40T)***:Mouse hepatic cells were isolated from mouse liver tissue fragments from 3-5 d old Kunming mice by the direct collagenase digestion method and cultured in *** pRSV-T plasmid was transfected into mouse hepatic cells to establish an SV40LT-immortalized mouse hepatic cell *** SV40LT-immortalized mouse hepatic cells were identified and transfected with the pHBV1.3 *** levels of hepatitis B surface antigen(HBsAg)and hepatitis B e antigen(HBeAg)in the supernatant were determined by an electrochemiluminescence immunoassay at 24,48,72 and 96 h after *** expressions of HBsAg and hepatitis B c antigen(HBcAg)in the cells were investigated by indirect immunofluorescence *** presence of HBV DNA replication intermediates in the transfected cells and viral particles in the supernatant of the transfected cell cultures was monitored using the Southern hybridization assay and transmission electronic microscopy,***:The pRSV-T plasmid was used to immortalize mouse hepatocytes and an SV40LT-immortalized mouse hepatic cell line was successfully ***40LT-immortalized mouse hepatic cells have the same morphology and growth characteristics as primary mouse hepatic cells can be subcultured and produce albumin and cytokeratin-18 in *** mouse hepatic cells did not show the characteristics of tumor cells,as alpha-fetoprotein levels were comparable(0.58±0.37 vs 0.61±0.31,P=0.37).SV40LTimmortalized mouse hepatic cells were then transfected with the pHBV1.3 plasmid,and it was found that the HBV genome replicated in SV40LT-immortalized mouse hepatic *** levels of HBsAg and HBeAg continuously increased in the supernatant after the transfection of pHBV1.3,and began to decrease 72 h after *** expressions of HBsAg and

关键词： SV40 T-antigen Mouse hepatic cell Hepatitis B virus 1.3-fold genome plasmids immortalized Liposomes Transfection

来源：

维普期刊数据库评论

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

Construction and identification of immortalized rat pulp stem cells

Construction and identification of immortalized rat pulp ste...

引用

中华口腔医学会第14次全国口腔医学学术会议（2012年会）

作者： Pang Xin,School of Stomatology Wuhan University,Wuhan,430000

Objective To construct and identify an immortalization system of rat pulp stem cell(rDPSC).Methods By using lentivirus-mediated gene transfection technology,SV40Tag was transfected in *** selection with GCV was used to obtain positive transfected *** the expression of SV40Tag in positive transfected cells by *** the limiting dilution method,the monoclonal cell of positive transfected cells was *** monaclonal cells were continuous passage cultured to calculate population doubling time,observe the cellular morphology and proliferative *** the 30th passage cell growth curve and analysis the differentiation *** We successfully selected the monoclonal cell from positive transfected *** monoclonal cells were positive for SV40Tag gene integration proved by *** monoclonal cells had been cultured for more than 50 *** immortalized rDPSC were polygonal-shaped, similar to primarily *** population doubling time of immortalized rDPSC was about half of the primarily *** 50th passage immortalized rDPSC still maintained a strong proliferation capacity and keep a multiple differentiation potential. Conclusion The immortalized rDPSC was successfully constructed and indentified.

关键词： immortalized rDPSC SV40Tag

来源： cnki会议评论

在线全文

cnki会议

学校读者我要写书评

暂无评论

Spontaneously immortalized B lymphocytes from patients with AIDS-related lymphoma are diagnostic and prognostic biomarkers

Spontaneously Immortalized B lymphocytes from patients with ...

引用

第十三届全国免疫学学术大会

作者： Ke Zhuang Yongxi Zhang Zhigao Xu Xiqiu Xu Fengzhen Meng Xiaoying Qi Yong Xiong Deying Guo Xien Gui ABSL-III Laboratory at the Center for Animal Experiment Wuhan University Department of Infectious Diseases Zhongnan Hospital of Wuhan University Department of Pathology Zhongnan Hospital of Wuhan University Basic Medical College of Wuhan University School of Basic Medicine (Shenzhen) Sun Yat-sen University

A valuable marker derived from patients with AIDS-related lymphoma(ARL) is needed for auxiliary diagnosis and prognosis. In this study, we generated spontaneously immortalized B lymphocyte cell(SIBC) lines by culturing peripheral blood mononuclear cells derived from 14 ARL patients and evaluated their clinical significance for auxiliary diagnosis and prognosis. From 10 pre-chemotherapy patients, a total of 5 new cell lines grew spontaneously without Cys A and EBV from five patients who showed a relapse and refractory ARL afterwards, presenting mainly memory B-cell markers(CD3-CD20+CD27+). However, SIBC lines could not be generated from healthy individuals(n=24). Among 8 post-chemotherapy patients, only 2 SIBC lines were generated from two ARL patients who had severe symptoms with multiple organs involved and died soon after chemotherapy;the other 6 ARL patients without SIBC lines survived for 1-5 years. Moreover, All SIBC lines exhibited monoclonal Ig gene rearrangement and presented a high level of cellular EBV DNA. These findings suggest that the SIBC lines are representative of the malignant B-lymphocytes circulating in ARL patients and might originate from the lymphoma tissue of ARL patients. These data provide evidence of a potentially valuable auxiliary diagnostic and prognostic biomarker for patients with ARL.

关键词： Spontaneous immortalized B cell AIDS-related lymphoma ARL

来源： cnki会议评论

在线全文

cnki会议

学校读者我要写书评

暂无评论

欢迎您,

建议与咨询留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

时间限定

文献类型

馆藏选择

核心期刊

语言

文献类型

帮助

文字说明：

检索规则说明：

检索范例：

分类表

所选分类

限定检索结果

文献类型

馆藏范围

日期分布

学科分类号

主题

机构

作者

语言

在线全文

在线全文

在线全文

在线全文

请选择保存的检索档案：

请选择收藏分类：

通借通还

欢迎您,

建议与咨询 留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

时间限定

文献类型

馆藏选择

核心期刊

语言

文献类型

帮助

文字说明：

检索规则说明：

检索范例：

分类表

所选分类

限定检索结果

文献类型

馆藏范围

日期分布

学科分类号

主题

机构

作者

语言

在线全文

在线全文

在线全文

在线全文

请选择保存的检索档案： 新增检索档案 确定 取消

请选择收藏分类： 新增自定义分类 确定 取消

通借通还

建议与咨询留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

请选择保存的检索档案：

请选择收藏分类：