检索结果-南通市图书馆

DNA Extraction from Formalin-fixed and Paraffin-embedded Tissues by Triton X-100 for Effective Amplification of EGFR Gene by Polymerase Chain Reaction

引用

Chemical Research in Chinese Universities 2012年第4期28卷 662-665页

作者： WANG Xiao-feng DU zhen-wu WU Meil zhang Yu-cheng JIANG Yang zhang gui-zhen Central Laboratory China-Japan Union Hospital of Jilin UniversityChangchun 130033P.R.China Dental Department China-Japan Union Hospital of Jilin UniversityChangchun 130033P.R.China Department of Rectum Colon Surgery China-Japan Union Hospital of Jilin UniversityChangchun 130033P.R.China

For first-line non-small-cell lung cancer（NSCLC） therapy,detecting mutation status of the epidermal growth factor receptor（EGFR） gene constitutes a prudent test to identify patients who are most likely to benefit from EGFR-tyrosine kinase inhibitor（TKI） ***,the material for detecting EGFR gene mutation status mainly comes from formalin-fixed and paraffin-embedded（FFPE） *** extraction from FFPE and the amplification of EGFR gene by polymerase chain reaction（PCR） are two key steps for detecting EGFR gene *** showed a simple method of DNA extraction from FFPE tissues for the effective amplification of EGFR *** DNA from the FFPE tissues of NSCLC patients with 1% Triton X-100（pH=10.0） was performed by heating at 95 °C for 30 ***,a commercial kit was used to extract DNA from the same FFPE tissues of NSCLC patients for *** extracted products were used as template for amplifying the exons 18,19,20 and 21 of EGFR by PCR for different amplified *** show that DNA fragment size extracted from FFPE tissues with 1% Triton X was about 250―500 base pairs（bp）.However,DNA fragment size extracted from FFPE tissues via commercial kit was about from several hundreds to several thousands *** DNA yield extracted from FFPE tissues with 1% Triton X was larger than that via commercial *** about 500 bp fragment,four exons of EGFR could not be amplified more efficiently from extracted DNA with 1% Triton X than with commercial ***,for about 200 bp *** simple and non-laborious protocol could successfully be used to extract DNA from FFPE tissue for the amplification of EGFR gene by PCR,further screening of EGFR gene mutation and facilitating the molecular analysis of a large number of FFPE tissues from NSCLC patients.

关键词： EGFR gene amplification DNA extraction Formalin-fixed and paraffin-embedded tissue Non-small-cell lung cancer

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

Protective Effect of Angiotensin(1-7) on Silicotic Fibrosis in Rats

引用

Biomedical and Environmental Sciences 2019年第6期32卷 419-426页

作者： zhang Bo Nan XU Hong GAO Xue Min zhang gui zhen zhang Xin YANG Fang School of Public Health North China University of Science and Technology Tangshan 063210 Hebei China Hebei Key Laboratory for Organ Fibrosis Medical Research Center North China University of Science and Technology Tangshan 063210 Hebei China Hebei Key Laboratory for Chronic Diseases Tangshan Key Laboratory for Clinical and Basic Research on Chronic Diseases School of Basic Medical Sciences North China University of Science and Technology Tangshan 063210 Hebei China Basic Medical College Hebei Medical University Shijiazhuang 050017 Hebei China

Objective Silicosis, caused by inhalation of silica dust, is the most serious occupational disease in China and the aim of present study was to explore the protective effect of Ang(1-7) on silicotic fibrosis and myofibroblast differentiation induced by Ang II. Methods HOPE-MED 8050 exposure control apparatus was used to establish the rat silicosis model. Pathological changes and collagen deposition of the lung tissue were examined by H.E. and VG staining, respectively. The localizations of ACE2 and α-smooth muscle actin(α-SMA) in the lung were detected by immunohistochemistry. Expression levels of collagen type I, α-SMA, ACE2, and Mas in the lung tissue and fibroblasts were examined by western blot. Levels of ACE2, Ang(1-7), and Ang II in serum were determined by ELISA. Co-localization of ACE2 and α-SMA in fibroblasts was detected by immunofluorescence. Results Ang(1-7) induced pathological changes and enhanced collagen deposition in vivo. Ang(1-7) decreased the expressions of collagen type I and α-SMA and increased the expressions of ACE2 and Mas in the silicotic rat lung tissue and fibroblasts stimulated by Ang II. Ang(1-7) increased the levels of ACE2 and Ang(1-7) and decreased the level of Ang II in silicotic rat serum. A779 enhanced the protective effect of Ang(1-7) in fibroblasts stimulated by Ang II. Conclusion Ang(1-7) exerted protective effect on silicotic fibrosis and myofibroblast differentiation induced by Ang II by regulating ACE2-Ang(1-7)-Mas axis.

关键词： Silicosis Ang II Myofibroblasts Ang(1-7) A779

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

Improved Allele-specific Polymerase Chain Reaction for Single Nucleotide Polymorphism Genotyping

引用

Chemical Research in Chinese Universities 2010年第2期26卷 259-262页

作者： WU Mei DU zhen-wu LIU Jia-nan SONG Yang WANG Ya-li zhang gui-zhen Department of Central Research China-Japan Union Hospital of Jilin University Changchun 130033 P. R. China

An improved allele-specific PCR（AS-PCR） approach was applied to investigating -55C/T polymorphism in promoter region of the uncoupling protein 3（UCP3）gene. AS-PCR is a competitive PCR method which is based on positioning the 3＇ base of a PCR primer to match one single nucleotide polymorphism（SNP） allele and accurately extend only the correctly matched primer. But it is limited in use because of its poor specificity. In this study, we improved the specificity of AS-PCR by introducing additional mismatch at the penultimate base of 3＇ end of AS-PCR primer in combination with decreasing the level of dNTP in the reaction mixture. Sensitivity, specificity and reliability of this method were assessed for both simple plasmid model and complex human genomic SNP targets. The -55C/T（rs1800849） polymorphisrn of the UCP3 gene was analyzed via this AS-PCR and restriction fragment length polymorphism（RFLP）, the latter was used as a gold standard. The results suggest that the increase in AS-PCR discrimination with this method should facilitate the use of this simple, rapid, and inexpensive technique for SNP genotyping application.

关键词： AS-PCR Polymorphism UCP3 SNP genotyping

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

Immunofluorescent Labeling of Human HepG2 Cells with CdTe Quantum Dot Probe Conjugated with Anti-pan CK MAb

引用

Chemical Research in Chinese Universities 2011年第2期27卷 277-281页

作者： SUI Yu-jie zhang gui-zhen WANG Qian WANG Ya-li WU Mei DU zhen-wu zhang Jie JIANG Ri-hua Central Laboratory China-Japan Union Hospital of Jilin University Changchun 130033 P. R. China Central Laboratory the Second Hospital of Jilin University Changchun 130041 P. R. China Department of Urinary Surgery China-Japan Union Hospital of Jilin University Changchun 130033 P. R. China College of Chemistry Jilin University Changchun 130012 P. R. China

A relatively sensitive, specific, and photostable method for the detection of cytokeratin of cancer cells via conjugation with cadmium telluride quantum dots（CdTe QDs） was described. Water soluble CdTe QDs were conjugated to anti-pan-cytokeratin（CK） monoclonal antibody（MAb） through coupling reagent [1-ethyl-3-（3-dimethyla- mino propyl）carbodiimide, EDC] and the conjugates were purified by dialysis. The expression of pan CK protein in HepG2 cells was observed by immunocytochemistry and direct immunofluorescence via QDs-Ab conjugates respectively. Fluorescence intensity and photostability of QDs were compared with those of FITC（fluorescein isothiocyanate）. The results show that the QDs-Ab conjugates recognized specifically pan CK protein in HepG2 cells. Compared with FITC, CdTe QDs had higher brightness and photostability without obvious photobleaching under continuous exciting light illumination for 30 min and after the placement at room temperature for 3 d. The results indicate that conjugates of CdTe quantum dot with anti-pan CK MAb can be used for labeling cancer cells derived from epithelial tissues, which provides the basis for the detection of circulating tumor cells（CTCs）.

关键词： Quantum dot Pan-cytokeratin Immunofluorescence labeling Fluorescence photostability

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

Effects of Micronutrients on Oxidative Stress and Islet Function in Type 1 Diabetes Mellitus Rats:Relationships to Th2 Type Cytokines,Interleukin-4,and Interluekin-10

引用

Chemical Research in Chinese Universities 2007年第6期23卷 701-704页

作者： zhang gui-zhen LI Mei-hua SONG Yang LIU Ting GAO Shen SUN Ying Department of Central Research The Third Clinical College Jilin University Changchun 130033 P. R. China Department of Allergy ＆ Clinical Immunology Faculty of Medicine Imperial College School National Heart ＆ Lung Institute Dovehouse Street London SW3 6LY UK

To observe the effects of the micronutrients on oxidative and autoimmune destruction of islets so as to prevent a person from the onset and development of type 1 Diabetes Mellitus （T1DM）, the interleukin-4 and interleukin-10 expressions of lymphocytes in peripheral blood and spleen of T1DM rats were determined by flow cytometry. GSH-Px activity and MDA level in the rats＇ pancreas were measured using biochemical methods. The insulin contents in serum and β cell insulin secret storage were tested by RIA and IHC, respectively. There was an increase in the percentages of IL-4 and IL-10 positive lymphocytes in the peripheral blood and spleen of the groups of rats supplemented with various combinations of micronutrients（p 〈0.01 and p 〈0.05, respectively） ; the blood glucose concentration decreased （p 〈 0. 05 ） ; both the functional β cell in islets and the insulin content in pancreatic tissue increased （p 〈 0. 05 and p 〈0. 01 ） ; the GSH-Px activity and MDA level of pancreas in the rats enhanced and decreased respectively（p 〈0. 01 and p 〈 0. 05）. The results suggest that micronutrients may alleviate the islet lesions by upregulating the expressions of IL-4 and IL-10 and lowering oxidative stress in diabetic rats .

关键词： Immanohistochemistry Oxidative stress Micronutrients T1 DM Interleukin-4 Interluekin-10

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

Association of UCP3,APN,and TNF-α Gene Polymorphisms with Type 2 Diabetes in a Population of Northern Chinese Han Patients

引用

Chemical Research in Chinese Universities 2012年第2期28卷 255-258页

作者： WANG Ling-ling DU zhen-wu LIU Jia-nan WU Mei SONG Yang JIANG Ri-hua zhang gui-zhen Central Research Department China-Japan Union Hospital of Jilin UniversityChangchun 130033P.R.China1 Department of Orthopedics China-Japan Union Hospital of Jilin UniversityChangchun 130033P.R.China1 Department of Neurology the Affiliated Hospital of Beihua UniversityJilin 132000P.R.China

We observed the polymorphism distribution and coaction of uncoupling protein 3（UCP3）-55C/T,adiponectin（APN）＋45T/G and tumor necrosis factor（TNF）-α-308G/A on the onset and development of T2DM in a Northern Chinese Han population of 213[100 type 2 diabete（T2DM） patients and 113 health control subjects] by polymerase chain reaction-restriction fragment length polymorphisum（PCR-RFLP） *** demonstrate the polymorphism of UCP3-55C/T,APN＋45T/G,and TNF-α-308G/A related to T2DM onset and *** the individuals carrying UCP3-55T,APN＋45G and TNF-α-308A allele had higher T2DM *** results are the first report to evaluate the association of the coaction of UCP3,APN,TNF-α genes polymorphism on T2DM risk and the susceptibility of T2DM in the Northern Chinese Han population.

关键词： Uncoupling protein 3(UCP3) Adiponectin(APN) Tumor necrosis factor(TNF)-α Gene polymorphism Type 2 diabete(T2DM) risk

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

Effect of Regulation of HSV-tk Gene Expression and Tumor Killed Activity with a Single Tetracycline-regulatable Plasmid Vector on HeLa Cells

引用

Chemical Research in Chinese Universities 2009年第5期25卷 677-680页

作者： WANG Qian DU zhen-wu MA Qing-shan zhang Yu-cheng WU Xiao-dong YANG Shao-juan WANG Ya-li zhang gui-zhen Central Laboratory China-Japan Union Hospital Jilin University Changchun 130033 P. R. China Department of Urinary Surgery China-Japan Union Hospital Jilin University Changchun 130033 P. R. China School of Pharmaceutical Sciences The First Hospital Jilin University Changehun 130021 P. R. China Department of Pediatrics The First Hospital Jilin University Changehun 130021 P. R. China

To construct a single tetracycline-regulatable plasmid vector based on the double tetracycline-regulatable plasmid vector system for regulating HSV-tk gene expression so as to effectively kill HeLa cells. Two tetracycline operator（TetO2） was cloned into pcDNA3.1 and a cassette was made for a cytomegalovirus-type 2 tetracycline operator（CMV-TetO2） promoter, and the obtained vector was named pcDNA3.1-CMV-TetO2. Herpes simplex virus thymidine kinase（HSV-tk） gene and tetracycline repressor（TR） gene were cloned into pcDNA3.1-CMV-TetO2 and the two genes were linked with internal ribosome entry site（IRES） to gain a vector named pcDNA3.1-CMV-TetO2- HSV-tk-IRES-TR. The HeLa cells were stablly transfected with pcDNA3.1-CMV-TetO2-HSV-tk-IRES-TR plasmid. The expression of HSV-tk and TR were detected by RToPCR, the tumorcidal activity of HSV-tk/GCV was determined by MTT assay. In Hela cells transfected with the above plasmid vector, HSV-tk gene and TR gene can be expressed lowly and the concentration of GCV producing a 50% decrease in cell viability was about 50 lag/mL without adding deoxycycline; in contrast, the expessions of HSV-tk gene and TR gene increased significantly and the concentration of GCV producing a 50% decrease in cell viability was about 5 μg/mL with adding deoxycycline. Therefore tetracycline can regulate the expression and tumorcidal activity of HSV-tk gene in HeLa cells with this single plasmid vector.

关键词： HSV-tk gene Tetracycline HeLa cells Gene expression

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

进口再生橡胶的固体废物属性鉴别方法探讨

引用

中国口岸科学技术 2024年第4期6卷 34-40页

作者：蒋一昕袁敏朱海欧吴璟张彰崔祥慧张桂珍南京海关工业产品检测中心南京210019 南京金检检验有限公司南京210019

为保证进口再生橡胶原料质量,防止硫化橡胶废碎料等属于我国禁止进口的固体废物以再生橡胶原料名义进口,因此需对进口再生橡胶原料实施固体废物属性鉴别。目前针对进口再生橡胶的固体废物属性鉴别尚无统一的标准方法。本文通过外观检查... 详细信息

为保证进口再生橡胶原料质量,防止硫化橡胶废碎料等属于我国禁止进口的固体废物以再生橡胶原料名义进口,因此需对进口再生橡胶原料实施固体废物属性鉴别。目前针对进口再生橡胶的固体废物属性鉴别尚无统一的标准方法。本文通过外观检查、红外光谱分析、裂解气相色谱-质谱和再生橡胶的性能指标测试等手段,考察进口再生橡胶原料的均匀程度,确定回收橡胶的种类,测试其性能参数,结合生产工艺和原料来源等因素进行综合判断,为准确鉴别再生橡胶固体废物属性提供了参考依据。

关键词：再生橡胶固体废物属性鉴别

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

Fluorosis Caused Cellular Apoptosis and Oxidative Stress of Rat Kidneys

引用

Chemical Research in Chinese Universities 2013年第2期29卷 263-269页

作者： SONG Yang WANG Jin-cheng XU Hui DU zhen-wu zhang gui-zhen SELIM Hamid Abdu LI Guang-sheng WANG Qing GAO Zhong-li Department of Orthopedics P. R. China Central Laboratory China-Japan Union Hospital of Jilin UniversityChangchun 130033 P. R. China Pathological Laboratory Institute of Endemic Disease of Jilin University Changchun 130021 P. R. China

As the strongest electronegative element, fluorine can stimulate the production of superoxide radicals in cells. In view of the important roles of kidneys in bone metabolism, the authors analyzed the quantitative pathomor- phological characteristics of renal damage and the potential cellular apoptosis and oxidative stress mechanisms in rats treated with excessive fluoride. Wistar rats were exposed to 50 mg F （110.5 mg NaF）/L, 100 mg F-（221.0 mg NaF）/L and 150 mg F-（331.5 mg NaF）/L in drinking water for 70 and 140 d, respectively. Microscope with image analysis was used to quantitate pathomorphological changes in renal tissues of the rats. Reactive oxygen species（ROS）, the cell cycle and apoptosis of renal cells were measured by flow cytometry and TUNEL technique（terminal deoxynuc- leotidyl transferase dUTP nick end labeling）, respectively. The ion concentrations in serum and renal functional pa- rameters were detected by automatic biochemical analyzer. Quantitative analysis results demonstrate the expanded Bowman＇s space of glomerulus and obvious dilatation of renal tubule. TUNEL technique revealed that NBT/BCIP （nitro blue tetrazolium/5-bromo-4-chloro-3＇-indolylphosphate, p-toluidine salt）-staining positive apoptotic cells se- lectively located in medullocortical junction areas. The data suggest that renal damage in chronic fluorostic rats is associated with the cellular apoptosis and oxidative stress.

关键词： Fluorosis Apoptosis Oxidative stress

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

MicroRNA-mediated Silencing of RhoC Inhibits Tumor Invasion and Increases Chemosensitivity to Paclitaxel in SKOV3 Cells in vitro

引用

Chemical Research in Chinese Universities 2011年第1期27卷 70-74页

作者： PAN Ying DU zhen-wu LENG Wei-chun ZHOU Jia-wen WANG Ying-jian SHENG Min-jia WANG Jun-rong zhang gui-zhen China-Japan Union Hospital of Jilin University Changchun 130031 P. R. China

RhoC is a member of the Ras-homologous family of genes which are implicated in tumorigenesis and tumor progression. Up-regulation of RhoC is associated with tumor progression in ovarian carcinoma and RhoC is significantly correlated with the invasive capability of ovarian cancer cell lines in vitro, We developed a system that blocks RhoC in the human ovarian cancer SKOV3 cells using specific MicroRNA（miRNA） interference. By transfecting SKOV3 cells with the plasmid vector to express specific MiRNA that targets human RhoC, we were able to establish a stable clone in which RhoC expression was significantly downregulated. This resulted in the decreased invasive potential of SKOV3 cells as well as increased chemosensitivity to paclitaxel. RhoC involves in invasion and chemosensitivity of SKOV3, indicating that RhoC may be a promising therapeutic target for ovarian cancer.

关键词： RhoC gene miRNA SKOV3 cell Cell invasion Paclitaxel

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

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

在线全文

在线全文

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

在线全文

在线全文

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：