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Hepatitis C virus in human B lymphocytes transformed by Epstein-Barr virus in vitro by in situ reverse transcriptase-polymerase chain reaction 被引量:11
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作者 Ji Lin Cheng Bao Ling Liu Yi Zhang Wen Bin Tong Zheng Yan Bai Fang Feng Institute of Hepatology,Peoples Hospital,Medical Center of Beijing University,Beijing 10(X)44,China 《World Journal of Gastroenterology》 SCIE CAS CSCD 2001年第3期370-375,共6页
AIM To study persistence and replication ofheltitis C virus (HCV) in patients' peripheralblood mononuclear cells (PBMC) cultured invitro.METHODS Epstein-Barr virus (EBV) was usedto transform the hepatitis C virus ... AIM To study persistence and replication ofheltitis C virus (HCV) in patients' peripheralblood mononuclear cells (PBMC) cultured invitro.METHODS Epstein-Barr virus (EBV) was usedto transform the hepatitis C virus from a HCVpositive patient to permanent lymphoblastoidcell lines (LCL). Positive and negative HCV RNAstrands of the cultured cells and growth mediawere detected by reverse transcriptase-polymerase chain reaction ( RT-PCR ) eachmonth. Core and NS5 proteins of HCV werefurther tested using immunohistochemical SPmethod and in situ RT-PCR.RESULTS HCV RNA positive strands wereconsistently detected the cultured cells for oneyear. The negative-strand RNA in LCL cells andthe positive-strand RNA in supernatants wereobserved intermittently. Immunohistochemicalresults medicated expression of HCV NS3 and Cproteins in LCL cytoplasm mostly. The positivesignal of PCR product was dark blue and mainlylocalized to the LCL cytoplasm. The RT-PCRsignal was eliminated by overnight RNasedigestion but not DNase digestion.CONCLUSION HCV may exist and remainfunctional in a cultured cell line for a longperiod. 展开更多
关键词 HEPATITIS C-like viruses HERPES virus 4 HUMAN B-LYMPHOCYTES cellscultured
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Detection of hepatocellular carcinoma cells in the peripheral blood with reverse--transcription polymerase chain reaction
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作者 房殿春 刘为纹 +1 位作者 罗元辉 鲁荣 《Journal of Medical Colleges of PLA(China)》 CAS 1998年第2期93-96,共4页
In order to detect circulating cells of hepatocellular carcinoma(HCC) in the peripheral blood with reverse transcripition polymerase chain reaction (RT-PCR ), alpha-fetoprotein (AFP ) mRNA was tested in the blood samp... In order to detect circulating cells of hepatocellular carcinoma(HCC) in the peripheral blood with reverse transcripition polymerase chain reaction (RT-PCR ), alpha-fetoprotein (AFP ) mRNA was tested in the blood samples of 113 cases of HCC and 69 controls (including 30 cases of liver cirrhosis, 9 cases of metastatic liver cancer and 30 normal subjects). 20/43 (46. 5% ) cases of HCC and 2/30 (6. 7% ) cases of liver cirrhosis are positive and the cases of nletastatic liver cancer and normal controls were negative for human AFP(hAFP) rnRNA. The presence of hAFP mRNA in the peripheral blood seems to be correlated with intrahepatic and distant nletastasls of HCC and portal vein thrombosis. It is concluded that the presence of hAFP mRNA in the peripheral hloocl is an indicator of circulating HCC cells and can be used to diagnose the rnetastasisof HCC through henlatogenous route and RT-PCR amplification of hAFP mRNA is a sensitive and specificprocedure for detecting circulating cells of HCC. 展开更多
关键词 hepatocellular carcinoma circulating cells ALPHA-FETOPROTEIN reverse transcription-polymerase chain reaction mRNA
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SEQUENCE ANALYSIS OF THE NS5 REGION OF GBVC/HGV AND DETECTION OF THE VIRUS BY REVERSE TRANSCRIPTASE PCR
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作者 陶其敏 常锦红 +3 位作者 魏来 杜绍财 王豪 孙焱 《Chinese Medical Sciences Journal》 CAS CSCD 1998年第4期221-224,共4页
INTRODUCTIONAfterreliableassaysforthedetectionofhepati-tisCvirus(HCV)andEvirusinfectionbecameavail-able,itwa... INTRODUCTIONAfterreliableassaysforthedetectionofhepati-tisCvirus(HCV)andEvirusinfectionbecameavail-able,itwasevidentthatmosto... 展开更多
关键词 GBV-C HGV NS5基因 基因序列分析 RT-PCR 病毒性肝炎
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新型冠状病毒Omicron I1566V突变位点MS2噬菌体病毒样颗粒的构建
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作者 杨静远 李永鑫 +3 位作者 史茜 刘春燕 梁梦洁 张新 《检验医学与临床》 CAS 2024年第8期1030-1034,共5页
目的构建新型冠状病毒Omicron I1566V突变位点MS2噬菌体病毒样颗粒。方法选取并合成带有6×His标签的MS2噬菌体的衣壳蛋白(CP)、成熟蛋白(A蛋白)及Omicron I1566V突变基因序列,插入pACYCDuet-1质粒构建重组载体,通过原核系统诱导表... 目的构建新型冠状病毒Omicron I1566V突变位点MS2噬菌体病毒样颗粒。方法选取并合成带有6×His标签的MS2噬菌体的衣壳蛋白(CP)、成熟蛋白(A蛋白)及Omicron I1566V突变基因序列,插入pACYCDuet-1质粒构建重组载体,通过原核系统诱导表达目的蛋白,纯化重组蛋白后利用透射电镜对蛋白质进行物理表征,最后通过反转录聚合酶链反应(RT-PCR)检测病毒样颗粒的热稳定性及耐核酸酶水解能力。结果成功构建包含有6×His标签的CP、A蛋白和Omicron I1566V突变基因序列的重组载体,经限制性内切酶BamHⅠ和KpnⅠ酶切鉴定和测序验证,结果均与预期相符。经诱导并纯化后,通过电镜观察到了大小均匀、直径为23~28 nm的病毒样颗粒,该病毒样颗粒经核酸酶消化后可在37℃条件下稳定储存20 d以上。结论该研究成功利用MS2噬菌体的CP和A蛋白构建了Omicron I1566V突变位点病毒样颗粒,为该突变位点的RT-PCR检测体系提供了可靠的质量保障。 展开更多
关键词 新型冠状病毒变异株 Omicron I1566V突变位点 反转录聚合酶链反应 MS2噬菌体 病毒样颗粒
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Cytopathological evaluations combined RNA and protein analyses on defined cell regions using single frozen tissue block 被引量:10
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作者 HONG LI, XIAO YAN CHEN, QING You KONG, JIA LIULaboratory of Cell Biology and Molecular Genetics,College of Basic Medical Sciences, Dalian Medical University, Dalian 116027, China 《Cell Research》 SCIE CAS CSCD 2002年第2期117-121,共5页
The co-existence of multiple cell components in tissue samples is the main obstacle for precise molecular evaluation on defined cell types. Based on morphological examination, we developed an efficient approach for pa... The co-existence of multiple cell components in tissue samples is the main obstacle for precise molecular evaluation on defined cell types. Based on morphological examination, we developed an efficient approach for paralleled RNA and protein isolations from an identical histological region in frozen tissue section.The RNA and protein samples prepared were sufficient for RT-PCR and Western blot analyses, and the results obtained were well coincident each other as well as with the corresponding parameters revealed from immunohistochemical examinations. By this way, the sampling problem caused by cell-cross contamination can be largely avoided, committing the experimental data more specific to a defined cell type. These novel methods thus allow us to use single tissue block for a comprehensive study by integration of conventional cytological evaluations with nucleic acid and protein analyses. 展开更多
关键词 RNA and protein preparation defined HISTOLOGICAL region SERIAL FROZEN section gene expression.
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HCV-RNA positivity in peripheral blood mononuclear cells of patients with chronic HCV-infection: does it really mean viral replication? 被引量:29
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作者 Volker Meier Sabine Mihm +1 位作者 Perdita Wietzke-Braun Guliano Ramadori 《World Journal of Gastroenterology》 SCIE CAS CSCD 2001年第2期228-234,共7页
AIM To analyze the association of HCV-RNA with peripheral blood mononuclear cells (PBMC)and to answer the question whether HCV-RNA positivity in PBMC is due to viral replication,METHODS HCV-RNA was monitored in seruma... AIM To analyze the association of HCV-RNA with peripheral blood mononuclear cells (PBMC)and to answer the question whether HCV-RNA positivity in PBMC is due to viral replication,METHODS HCV-RNA was monitored in serumand PBMC preparations from 15 patients with chronic HCV infection before, during and after an IFN-α therapy using a nested RT/ PCRtechnique. In a second approach, PBMC from healthy donors were incubated in HCV positive plasma.RESULTS In the IFN-α responding patients,HCV-RNA disappeared first from total RNApreparations of PBMC and then from serum. In contrast, in relapsing patients, HCV-RNAreappeared first in serum and then in PBMC. A quantitative analysis of the HCV-RNAconcentration in serum was performed before and after transition from detectable to nondetectable HCV-RNA in PBMC-RNA and vice versa. When HCV-RNA was detectable in PBMCpreparations, the HCV concentration in serum was significantly higher than the serum HCV-RNA concentration when HCV-RNA in PBMC was not detectable. Furthermore, at no time during the observation period was HCV specific RNA observed in PBMC, if HCV-RNA in serum was under the detection limit. Incubation of PBMCfrom healthy donors with several dilutions of HCV positive plasma for two hours showed a concentration-dependent PCR-positivity for HCV-RNA in reisolated PBMC.CONCLUSION The detectability of HCV-RNA in total RNA from PBMC seems to depend on the HCV concentration in serum. Contamination or passive adsorption by circulating virus could be the reason for detection of HCV-RNA in PBMCpreparations of chronically infected patients. 展开更多
关键词 HEPATITIS C-like viruses HEPATITIS C CHRONIC RNA viral/blood virus replication MONOCYTES interferon alpha/therapeutic use polymerase chain reaction
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Expressions of ICAM-1 and its mRNA in sera and tissues of patients with hepatocellular carcinoma 被引量:14
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作者 Jing Xu1 Ming Hui Mei1 +3 位作者 Si En Zeng2 Qing Fen Shi3 Yong Ming Liu4 Li Ling Qin3 1Department of Hepatobiliary Surgery2Department of Pathology3Institute of Hepatobiliary Surgery4Department of Biochemistry, Guilin 541001, Guangxi Province, China 《World Journal of Gastroenterology》 SCIE CAS CSCD 2001年第1期120-125,共6页
INTRODUCTIONThe increased expression of ICAM-1 on a widerange of cells and in the sera of patients withmalignancies, chronic liver diseases andinflammation diseases has been described since thelate 1980s[1-22]. Recent... INTRODUCTIONThe increased expression of ICAM-1 on a widerange of cells and in the sera of patients withmalignancies, chronic liver diseases andinflammation diseases has been described since thelate 1980s[1-22]. Recently rapid progress in studieson expression of ICAM-1 in patients withhepatocellular carcinoma ( HCC ) have beenachieved, including clinical and experimentalresearches[23-31]. 展开更多
关键词 LIVER neoplasms/diagnosis intercellular adhesion molecule-1 RNA messenger alpha-fetoprotein immunohistochemistry polymerase chain reaction radioimmunoassay CLC Number:R57 Document ID:A INTRODUCTIONThe increased EXPRESSION of ICAM-1 on a widerange of cells and in the SERA of PATIENTS withmalignancies chronic LIVER DISEASES andinflammation DISEASES has been described since thelate 1980s[1-22]. Recently rapid progress in studieson EXPRESSION of ICAM-1 in PATIENTS withhepatocellular carcinoma ( HCC ) have beenachieved including clinical and experimentalresearches[23-31].……
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Expression of mucosal addressin cell adhesion molecule 1 on vascular endothelium of gastric mucosa in patients with nodular gastritis 被引量:13
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作者 HiroshiOhara HajimeIsomoto +9 位作者 Chun-YangWen ChiekoEjima MasahiroMurata MasanobuMiyazaki FuminaoTakeshima YoheiMizuta IkuoMurata TakehikoKoji HiroshiNagura ShigeruKohno 《World Journal of Gastroenterology》 SCIE CAS CSCD 2003年第12期2701-2705,共5页
AIM: The interaction of mucosal addressin cell adhesion molecule 1 (MAdCAM-1) with integrin α4β7 mediates lymphocyte recruitment into mucosa-associated lymphoid tissue (MALT). Nodular gastritis is characterized by a... AIM: The interaction of mucosal addressin cell adhesion molecule 1 (MAdCAM-1) with integrin α4β7 mediates lymphocyte recruitment into mucosa-associated lymphoid tissue (MALT). Nodular gastritis is characterized by a unique military pattern on endoscopy representing increased numbers of lymphoid follicles with germinal center, strongly associated with H pylori infection. The purpose of this study was to address the implication of the MAdCAM-1/integrin β7 pathway in NG.METHODS: We studied 17 patients with NG and H pylori infection and 19 H pylori-positive and 14 H pylori-negative controls. A biopsy sample was taken from the antrum and snap-frozen for immunohistochemical analysis of MAdCAM1 and integrin β7. In simultaneous viewing of serial sections,the percentage of MAdCAM-1-positive to von Willebrand factor-positive vessels was calculated. We also performed immunostaining with anti-CD20, CD4, CD8 and CD68 antibodies to determine the lymphocyte subsets coexpressing integrin β7.RESULTS: Vascular endothelial MAdCAM-1 expression was more enhanced in gastric mucosa with than without H pylori infection. Of note, the percentages of MAdCAM-1-positive vessels were significantly higher in the lamina propria of NG patients than in H pylori-positive controls. Strong expression of MAdCAM-1 was identified adjacent to lymphoid follicles and dense lymphoid aggregates. Integrin β7-expressing mononuclear cells, mainly composed of CD20 and CD4 lymphocytes, were associated with vessels lined with MAdCAM-1-expressing endothelium.CONCLUSION: Our results suggest that the MAdCAM-1/integrin α4β7 homing system may participate in gastric inflammation in response to H pylori-infection and contributes to MALT formation, typically leading to the development of NG. 展开更多
关键词 结节性胃炎 胃粘膜 细胞粘附因子-1 血管内皮素 淋巴细胞
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Expression of Transforming Growth Factor-β in Cultured Normal Human Lens Epithelia Cells 被引量:2
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作者 黄渝侃 魏厚仁 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2004年第3期289-291,共3页
Summary: In order to investigate whether cultured normal human lens epithelial cells (LEC) express transforming growth factor β (TGF-β), reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemic... Summary: In order to investigate whether cultured normal human lens epithelial cells (LEC) express transforming growth factor β (TGF-β), reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemical methods were used for detection of TGF-β mRNA and protein in cultured normal human LEC. The results showed that a single RT-PCR amplified product about 310bp was obtained, and the sequence was homologous to the known sequence. TGF-β immunostain was positive in the plasma of LEC. It was suggested that normal human LEC could produce TGF-β, and LEC could be affected by TGF-β through autocrine action. 展开更多
关键词 LENS cell culture transforming growth factor reverse transcriptase-polymerase chain reaction IMMUNOHISTOCHEMISTRY
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Interleukin-1 beta up-regulates tissue inhibitor of matrix metalloproteinase-1 mRNA and phosphorylation of c-jun N-terminal kinase and p38 in hepatic stellate cells 被引量:22
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作者 Ya-Ping Zhang Xi-Xian Yao Xia Zhao 《World Journal of Gastroenterology》 SCIE CAS CSCD 2006年第9期1392-1396,共5页
瞄准:学习在 interleukin-1beta (IL-1beta ) 之间的关系矩阵 metalloproteinase-1 (TIMMP-1 ) 的起来调整的织物禁止者 mRNA 表示和两 c-jun N 终端激酶(JNK ) 和在老鼠的 p38 的磷酸化肝的星形细胞(HSC ) 。方法:RT-PCR 被执行在老... 瞄准:学习在 interleukin-1beta (IL-1beta ) 之间的关系矩阵 metalloproteinase-1 (TIMMP-1 ) 的起来调整的织物禁止者 mRNA 表示和两 c-jun N 终端激酶(JNK ) 和在老鼠的 p38 的磷酸化肝的星形细胞(HSC ) 。方法:RT-PCR 被执行在老鼠 HSC 测量 TIMMP-1 mRNA 的表示。西方的污点被执行在老鼠 HSC 测量 IL-1beta-induced JNK 和 p38 活动。结果:TIMMP-1 mRNA 表示(1.191+/-0.079 ) 比在控制组(0.545+/-0.091 )(P【0.01 ) 为 24 h 是有 IL-1beta (10 ng/mL ) 的许多更高的术后疗法。IL-1beta 以一种时间依赖者方式激活 JNK 和 p38。在有为 0, 5, 15, 30, 60 和 120 min 的 IL-1beta 的刺激以后, JNK 活动分别地是 0.982+/-0.299,1.501+/-0.720, 2.133+/-0.882, 3.360+/-0.452, 2.181+/-0.789,和 1.385+/-0.368。在在 15 min (P【0.01 ) 的 JNK 活动有有效差量, 30 min (P【0.01 ) 和在 0 min 的与那相比的 60 min (P【0.01 ) 。p38 活动分别地是在 6 个次点(0, 5, 15, 30, 60 和 120 min ) 的 1.061+/-0.310,2.050+/-0.863, 2.380+/-0.573, 2.973+/-0.953, 2.421+/-0.793,和 1.755+/-0.433。在在 5 点的 p38 活动有有效差量 min ( P【0.05 ), 15 min ( P【0.01 ), 30 min ( P【0.01 )和在在 3 减少的 0 min.TIMMP-1 mRNA 表示 trended 的与那相比的 60 min ( P【0.01 )与 SP600125 的不同集中组织 pretreated ( 10 micromol/L , 1.022+/-0.113 ;20 micromol/L, 0.869+/-0.070;40 micromol/L, 0.666+/-0.123 ) 。他们的减少都是重要的(P【0.05, P【0.01, P【0.01 ) 与控制组相比(没有 SP600125 处理, 1.163+/-0.107 ) 。在其它, 3 与 SB203580 的不同集中组织 pretreated (10 micromol/L, 1.507+/-0.099;20 micromol/L, 1.698+/-0.107;40 micromol/L, 1.857+/-0.054 ) , TIMMP-1 mRNA 的表示增加了。他们的层次比在控制组的那些高(没有 SB203580 处理, 1.027+/-0.061 ) 与重要统计意义(P【0.01 ) 。结论:IL-1beta 在老鼠 HSC 由起来调整的 TIMMP-1 mRNA 表示在肝的纤维变性上有一个直接行动。JNK 和 p38 激活 mitogen 的蛋白质家族 ases (MAPK ) 涉及 IL-1beta-induced TIMMP-1 基因表示,并且在这进程起一个不同作用,显示 p38 和 JNK 小径合作地调停在老鼠 HSC 的 TIMP-1 mRNA 表示。 展开更多
关键词 白细胞介素-1 组织抑制 金属蛋白 磷酸化
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Comparative identification of Ca~(2+) channel expression in INS-1 and rat pancreatic β cells 被引量:1
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作者 Fei Li Zong-Ming Zhang 《World Journal of Gastroenterology》 SCIE CAS CSCD 2009年第24期3046-3050,共5页
AIM:To identify and compare the profile of Ca2+ channel subunit expression in INS-1 and rat pancreatic β cells. METHODS:The rat insulin-secreting INS-1 cell line was cultured in RPMI-1640 with Wistar rats employed as... AIM:To identify and compare the profile of Ca2+ channel subunit expression in INS-1 and rat pancreatic β cells. METHODS:The rat insulin-secreting INS-1 cell line was cultured in RPMI-1640 with Wistar rats employed as islet donors.Ca2+ channel subunit expression in INS-1 and isolated rat β cells were examined by reverse transcription polymerase chain reaction (RT-PCR).Absolute real-time quantitative PCR was performed in a Bio-Rad iQ5 Gradient Real Time PCR system and the data analyzed using an iQ5 system to identify the expression level of the Ca 2+ channel subunits. RESULTS:In INS-1 cells,the L-type Ca 2+ channel 1C subunit had the highest expression level and the TPRM2 subunit had the second highest expression. In rat β cells,the TPRC4 β subunit expression was dominant and the expression of the L-type 1C subunit exceeded the 1D subunit expression about two-fold. This result agreed with other studies,confirming the important role of the L-type 1C subunit in insulinsecreting cells,and suggested that non-voltageoperated Ca 2+ channels may have an important role in biphasic insulin secretion. CONCLUSION:Twelve major Ca 2+ channel subunit types were identified in INS-1 and rat β cells and significant differences were observed in the expression of certain subunits between these cells. 展开更多
关键词 L型钙通道 胰腺细胞 INS 大鼠 比较鉴别 CA 逆转录聚合酶链反应 胰岛素分泌
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Plasma Membrane Calcium ATPase Expression in Human Lens Epithelium Cell Lines 被引量:2
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作者 Hui Yang Dongmei Cui Junwen Zeng 《Eye Science》 CAS 2011年第1期35-43,共9页
Purpose:To study the expression of four plasma membrane calcium ATPase (PMCA) isoforms in human lens epithelium cell lines (HLE-B3 cells) both on mRNA and protein levels.Methods:Both total mRNA and membrane protein sa... Purpose:To study the expression of four plasma membrane calcium ATPase (PMCA) isoforms in human lens epithelium cell lines (HLE-B3 cells) both on mRNA and protein levels.Methods:Both total mRNA and membrane protein samples were collected,after HLE-B3 cells were cultured to 90% confluency.Reverse Transcription Polymerase Chain Reaction (RT-PCR) were used to detect mRNAs of PMCA isoform 1,2,3,and 4 by using corresponding PMCA isoform 1,2,3,and 4 primers.Western Blot analysis was employed to detect PMCA isoform 1,2,3,and 4 protein using corresponding anti-PMCA1,2,3,and 4 antibodies.Results:A 420 bp fragment was amplified with PMCA1 primer.A 550 bp fragment was amplified with PMCA2 primer.A 840 bp fragment was amplified with PMCA4 primer.No fragment was amplified with PMCA3 primer.Western Blotting confirmed that the expected ~153 kDa,~125 kDa and ~147 kDa protein were recognized by anti PMCA1,2 and 4 antibodies respectively.No protein was recognized by PMCA3 antibody.Conclusion:This is the first study showing only PMCA1,2,and 4 gene are expressed in HLE-B3 cells on both mRNA and protein level.PMCA3 is not expressed in HLE-B3 cells.The PMCA isoforms expression pattern in HLE-B3 cell lines is different from that in the lens of other species.PMCA2 may play a more important role over other isoforms. 展开更多
关键词 钙ATP酶 细胞系 晶状体 上皮 逆转录聚合酶链反应 WESTERN印迹 蛋白水平 mRNA
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The gene expression patterns of BMPR2,EP300,TGFβ2,and TNFAIP3 in B-Lymphoma cells 被引量:1
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作者 Dong-Mei He Hong Wu +3 位作者 Xiu-Li Wu Li Ding Ling Xu Yang-Qiu Li 《Cancer Biology & Medicine》 SCIE CAS CSCD 2014年第3期202-207,共6页
Objective: The results of a previous study showed that a clear dysregulation was evident in the global gene expression of the BCL11A-suppressed B-lymphoma cells. In this study, the bone morphogenetic protein receptor,... Objective: The results of a previous study showed that a clear dysregulation was evident in the global gene expression of the BCL11A-suppressed B-lymphoma cells. In this study, the bone morphogenetic protein receptor, type II(BMPR2), E1 A binding protein p300(EP300), transforming growth factor-β2(TGFβ2), and tumor necrosis factor, and alpha-induced protein 3(TNFAIP3) gene expression patterns in B-cell malignancies were studied. Methods: The relative expression levels of BMPR2, EP300, TGFβ2, and TNFAIP3 mRNA in B-lymphoma cell lines, myeloid cell lines, as well as in cells from healthy volunteers, were determined by real-time quantitative reverse transcriptpolymerase chain reaction(qRT-PCR) with SYBR Green Dye. Glyceraldehyde-3-phosphate dehydrogenase(GAPDH) was used as reference. Results: The expression level of TGFβ2 mRNA in B-lymphoma cell lines was significantly higher than those in the cells from the healthy control(P<0.05). However, the expression level of TNFAIP3 mRNA in B-malignant cells was significantly lower than that of the healthy control(P<0.05). The expression levels of BMPR2 and EP300 mRNA showed no significant difference between B-malignant cell lines and the healthy group(P>0.05). In B-lymphoma cell lines, correlation analyses revealed that the expression of BMPR2 and TNFAIP3(r=0.882, P=0.04) had significant positive relation. The expression levels of BMPR2, EP300, and TNFAIP3 mRNA in cell lines from myeloid leukemia were significantly lower than those in the cells from the healthy control(P<0.05). The expression levels of TGFβ2 mRNA showed no significant difference between myeloid leukemia cell lines and the healthy control or B-malignant cell lines(P>0.05). The expression levels of BMPR2, EP300, and TNFAIP3 mRNA in B-lymphoma cells were significantly higher than those of the myeloid leukemia cells(P<0.05).Conclusion: Different expression patterns of BMPR2, EP300, TGFβ2, and TNFAIP3 genes in B-lymphoma cells exist. 展开更多
关键词 白血病细胞系 B细胞淋巴瘤 基因表达模式 TGFΒ2 mRNA表达水平 甘油醛-3-磷酸脱氢酶 骨形态发生蛋白 恶性肿瘤
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Human dental pulp stem cells express many pluripotency regulators and differentiate into neuronal cells
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作者 Behnam Ebrahimi Mohammad Mehdi Yaghoobi +1 位作者 Ali Mohammadi Kamal-abadi Maryam Raoofn 《Neural Regeneration Research》 SCIE CAS CSCD 2011年第34期2666-2672,共7页
Stem cells were isolated from human dental pulp using an optimized method, in which pulp pieces were digested by enzymes and immobilized to enhance cell outgrowth. Stem cell marker expression was detected by reverse t... Stem cells were isolated from human dental pulp using an optimized method, in which pulp pieces were digested by enzymes and immobilized to enhance cell outgrowth. Stem cell marker expression was detected by reverse transcription-PCR (RT-PCR), and differentiation markers were detected by real-time quantitative RT-PCR and immunocytochemistry. Results showed that dental pulp stem cells actively expressed nanog, oct4, nucleostemin slain-l, jmjdla, jmjd2c, and cyclin DI. When stem cells were induced to differentiate into neurons, nucleostemin, nanog, and cyclin D1 expression significantly decreased, whereas expression of neuronal markers, such as microtubule associated protein-2 and neurofilament-heavy, significantly increased. These results suggested that stem cells exited a pluripotent state and entered a neuronal differentiation pathway. In addition, results demonstrated that human dental pulp serves as a reservoir of stem cells that express defined stem cell markers; these cells were easily isolated and were induced to differentiate towards a desired cell lineage. 展开更多
关键词 dental pulp stem cell neural regeneration neuronal differentiation PluriNet real-time quantitative reverse transcription polymerase chain reaction stem cell marker
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Transforming Growth Factor-β_2 Gene Cloning and Protein Expression in Human Trabecular Meshwork Cells
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作者 曹阳 魏厚仁 +1 位作者 笪邦红 李忠玉 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2003年第1期85-87,共3页
Whether cultured human trabecular meshwork cells express transforming growth factor-β 2 (TGF-β 2) messenger RNA (mRNA) and protein was investigated. Total RNA of 10 6 cultured human trabecular meshwork cells was ... Whether cultured human trabecular meshwork cells express transforming growth factor-β 2 (TGF-β 2) messenger RNA (mRNA) and protein was investigated. Total RNA of 10 6 cultured human trabecular meshwork cells was extracted with TRIZOL reagent, reverse transcriptase-polymerase chain reaction (RT-PCR) were used for detection of TGF-β 2 messenger RNA, and the PCR product was verified by sequencing. Immunohistochemical staining was used to detect TGF-β 2 protein. The results showed that a single RT-PCR amplified product was obtained, and the sequence was homologous to the known sequence. TGF-β 2 immunostaining was positive. It was concluded that trabecular meshwork cells could produce TGF-β 2 and contribute to the presence of TGF-β 2 in trabecular meshwork microenvironment as well as aqueous humor. Trabecular meshwork cells were affected by TGF-β 2 not only through paracrine, but also autocrine action. Whether abnormal changes in TGF-β 2 production contribute to the pathogenesis of primary open-angle glaucoma is worth further investigation. 展开更多
关键词 HUMAN trabecular meshwork cultured cells transforming growth factor reverse transcriptase-polymerase chain reaction IMMUNOHISTOCHEMISTRY
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Influence of all-trans retinoic acid(ATRA) on the expression of NANOG in Glioma cell lines
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作者 Liu Weixian Yi Fuxin 《重庆医学》 CAS CSCD 北大核心 2012年第7期625-629,F0002,共6页
Objective To investigate the effects of all-trans retinoic acid(ATRA) on the expression of NANOG in glioma cell lines.Methods Each cell line was divided into the experimental group which was treated with ATRA for 5 da... Objective To investigate the effects of all-trans retinoic acid(ATRA) on the expression of NANOG in glioma cell lines.Methods Each cell line was divided into the experimental group which was treated with ATRA for 5 days,and the control group which was cultured normally without ATRA treatment.Immunocytochemistry and RT-PCR were adopted to detect the expression of NANOG at protein and mRNA level among the three kinds of cell lines.Results Positive rates of NANOG protein in glioma cell lines SHG-44,U87 MG and U251 in control groups were(65.5±3.0)%,(64.8±8.0)% and(64.5±1.2)%,respectively,and the difference was not statistically significant(F=0.190,P=0.829).NANOG mRNA of the three cell lines in the relative content were 0.636 8±0.039 9,0.642 1±0.063 7,0.651 6±0.044 4,and the difference was not statistically significant(F=0.427,P=0.662).However,5 days after application of ATRA-induced NANOG protein in the three cell lines,the positive rates of NANOG protein of experimental groups were(36.5±7.3)%,(35.5±7.9)%,(35.2±6.1)%,respectively,compared with the control groups,the differences were statistically significant(FSHG-44=259.1,FU87=129.5,FU251= 431.8,PSHG-44=0.0,PU87=0.0,PU251=0.0),and the relative level of NANOG mRNA in these groups were 0.458 3±0.079 1,0.255 1±0.079 3 and 0.333 1±0.054 0,respectively,compared with the control groups,the difference was significant(FSHG-44=77.8,FU87=277.9,FU251=398.1,PSHG-44=0.0,PU87=0.0,PU251=0.0).Conclusion NANOG which highly expressed in glioma cell line SHG-44,U87 MG and U251 can be reduced by ATRA. 展开更多
关键词 《重庆医学》 期刊 编辑部 编辑工作
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Expression of Polt in tissues and cell lines of transitional cell carcino-ma
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作者 赵谦 金锡御 +7 位作者 杨劲 陈志文 宋波 曾益军 位全芳 季惠翔 李劲 周虎传 《Journal of Medical Colleges of PLA(China)》 CAS 2007年第6期337-340,共4页
Objective:To explore the expression of DNA polymerase iota in transitional cell carcinoma cells and tissues;Methods:RT-PCR was applied to detect the expression of polymerase iota in BIU87 and T24 cells,then the expres... Objective:To explore the expression of DNA polymerase iota in transitional cell carcinoma cells and tissues;Methods:RT-PCR was applied to detect the expression of polymerase iota in BIU87 and T24 cells,then the expression of polymerase iota was also detected in the same way in transitional cell car- cinoma which was derived from clinical bladder carcinoma and renal pelvic carcinoma.Results:The expres- sion of Poli was low in bladder normal membrana mucosa but significantly elevated in transitional cell car- cinoma cells.Compared with the expression of polymerase iota in bladder normal mucous membranes,the expression of polymerase iota was significantly increased in transitional cell carcinoma tissue(P<0.01) and associated with the grade of transitional cell carcinoma.Conclusion:The significantly increased ex- pression of polymerase iota may be associated with the generation and development of transitional cell car- cinoma,even with its high heterogenicity. 展开更多
关键词 细胞癌 损伤分析 DNA聚合酶 转录聚合酶链相互作用
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Comparison of phenotype characteristics of rat annulus fibrosus cells cultured on flexible silicone membrane and in plastic plate
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作者 郭志良 成敏 +3 位作者 曹国永 李华壮 滕海军 周跃 《Journal of Medical Colleges of PLA(China)》 CAS 2006年第2期112-116,共5页
Objective:To compare the phenotype characteristics of rat annulus fibrosus (AF) cells cultured on flexible silicone membranes and those in plastic plates. Methods:The morphology of AF cells cultured in different subst... Objective:To compare the phenotype characteristics of rat annulus fibrosus (AF) cells cultured on flexible silicone membranes and those in plastic plates. Methods:The morphology of AF cells cultured in different substrates was examined. Proteoglycan was stained by toluidine blue. Contents of collagen type I , collagen type II and aggrecan mRNAs were determined by reverse transcription-polymerase chain reaction (RT-PCR). The expression of integrin β1 was monitored by flow cytometry. By using propidium iodide (PI), the cell cycle in AF cells was analyzed. Cell adhesion to silicone membrane was also measured. Results:The AF cells cultured on different substrates were morphologically undistinguishable. Toluidine blue staining showed that there was also no difference between AF cells cultured on these 2 substrates. They still had the same expression levels of collagen type I , collagen type II , aggrecan mRNAs, and integrin β1. No significant difference was observed in the distribution of the cell cycle. AF cells grew well on silicone membrane. Conclusion: AF cells cultured on flexible silicone membrane maintain the stability of phenotype and may be appropriate for further studying the metabolic responses to mechanical stimuli at the cellular level. 展开更多
关键词 环状细胞 硅元素 塑料板 材料
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Expression of PinX1 and hTERT in basal cell carcinoma and their implications
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作者 Long Qin Jing Ge 《Oncology and Translational Medicine》 CAS 2015年第3期140-143,共4页
Objective This study aimed to investigate the expression and significance of PIN2/TERF1 interacting, telomerase inhibitor 1(Pin X1) and human telomerase reverse transcriptase(h TERT) in basal cell carcinoma(BCC). Meth... Objective This study aimed to investigate the expression and significance of PIN2/TERF1 interacting, telomerase inhibitor 1(Pin X1) and human telomerase reverse transcriptase(h TERT) in basal cell carcinoma(BCC). Methods Real-time polymerase chain reaction and immunohistochemistry were performed to quantify the m RNA expressions and integrated optical density(IOD), respectively, of Pin X1 and h TERT in BCC specimens(n = 30), as well as in normal skin specimens(n = 15). Results The m RNA expression level and IOD of Pin X1 in the BCC samples were both significantly lower than those in the control specimens(P < 0.05). Conversely, the m RNA expression level and IOD of h TERT in BCC were both significantly higher than that in the control samples(P < 0.05). The correlation between the expression levels of Pin X1 and h TERT showed no statistical significance(P > 0.05). Conclusion Downregulation of Pin X1 and upregulation of h TERT expression may be associated with the activation and maintenance of telomerases in the induction of BCC. 展开更多
关键词 肿瘤 诊断 临床 化疗 疗效
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Expression pattern of BIM,BCL-6,and c-MYC in adult B-cell acute lymphoblastic leukemia
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作者 Chanli Zheng Lin Xu +2 位作者 Yanjun Xie Dongmei He Yangqiu Li 《Oncology and Translational Medicine》 2017年第4期151-155,共5页
Objective We aimed to evaluate the expression pattern of the genes BIM, BCL-6, and c-MYC in adult patients at initial diagnosis of B-cell acute lymphoblastic leukemia(B-ALL).Methods Relative m RNA levels of BIM, BCL-6... Objective We aimed to evaluate the expression pattern of the genes BIM, BCL-6, and c-MYC in adult patients at initial diagnosis of B-cell acute lymphoblastic leukemia(B-ALL).Methods Relative m RNA levels of BIM, BCL-6, and c-MYC in peripheral blood mononuclear cells(PBMCs) from B-ALL patients were determined by quantitative reverse-transcription polymerase chain reaction(q RT-PCR) using SYBR Green dye. PBMCs from healthy volunteers served as a control. GAPDH was used as a reference gene.Results Relative expression of BIM, BCL-6, and c-MYC m RNA in B-ALL patients was significantly lower than in healthy controls(P < 0.05). Furthermore, this result was observed for both newly diagnosed B-ALL patients and those incomplete remission(CR)(P < 0.05). There were no statistically significant differences in the expression levels of BIM, BCL-6, and c-MYC between these B-ALL patient groups(P > 0.05). Spearman's rank correlation analyses revealed the expression level of BIM to be positively correlated with that of BCL-6 in B-ALL patients.Conclusion Expression of the genes BIM, BCL-6, and c-MYC is decreased in adult B-ALL patients. Moreover, the expression pattern of these genes may be similar in such patients at initial diagnosis and following CR. The expression characteristics of BIM, BCL-6, and c-MYC may constitute useful markers for the diagnosis of adult B-ALL. 展开更多
关键词 BIM BCL-6 C-MYC B cell acute LYMPHOBLASTIC leukemia (ALL) quantitative reverseTRANSCRIPTION polymerase chain reaction (qRT-PCR)
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