Qichang

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Fudan University

Shaobo

Zifang

Lirong

Xia

Ruijin Hospital

Rong

University of Hong Kong

Jun Jun

Wuhan University of Technology

Z. Zheng

Hangzhou Normal University

S Song

ShenZhen People’s Hospital

Wang

Qingdao University

Cooperative Institutions

Shanghai Jiao Tong University

Chinese Academy of Sciences

Fudan University

Second Military Medical University

Dalian Medical University

Chongqing University

Chinese Academy of Agricultural Sciences

Zhejiang University

Anhui Medical University

University of Hong Kong

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Two-dimensional gel electrophoresis analysis of the proteomes expressed in the human hepatoma cell line BEL-7404 and normal liver cell line L-02

中国科学通报：英文版 (2000)

Lirong Yu Wang Gaode Wu

Proteome analysis technology has been used extensively in conducting discovery research of biology and has become one of the most essential technologies in functional genomics. The proteomes of the human hepatoma cell line BEL-7404 and the normal human liver cell line L-02 have been separated by high resolution two-dimensional gel electrophoresis (2-DE) with immobilized pH gradient isoelectric focusing (IPG-IEF) in the first dimension and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) in the second dimension (IPG-DALT). The resulting images have been analyzed using 2-D analysis software. Quantitative analysis reveals that 7 protein spots are detected only in hepatoma BEL-7404 cells, 14 only in L-02 cells, and 78 protein spots show significant fluctuation in quantity in both cell lines (P<0.01). These protein spots have been displayed on a proteome differential expression map. Analysis for the reproducibility of 2-DE indicates that the positional variability in the IEF dimension is

Proteome

Immobilized pH gradient

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The effect of human papilloma virus type 16 E7 protein expression on growth of RMA cells in vitro and in vivo

中德临床肿瘤学杂志：英文版 (2010)

Shaobo Hu Qichang Zheng Zifang

这研究的目的是学习人的乳突淋瘤的效果的目的在 vitro 并且在 vivo 的 RMA 房间的生长上的病毒(HPV ) 类型 16 E7 蛋白质表示。带野类型 HPV 16 E7 的再结合向量 pcDNA3.1-E7 被定序识别的方法。再结合向量 pcDNA3.1-E7 是进由 liposome 的老鼠淋巴腺瘤房间线 RMA 的 transfected，并且 monoclonal 房间 transfected 被抗菌素 G418 sieving 和限制冲淡试金稳定地获得。RT-PCR 方法被用来在 RMA-E7 房间检测 HPV 16 E7 mRNA 的表示。RMA 细胞和在 vitro 有教养的 RMA-E7 细胞的生长被细胞计数 Kit-8 测试。RMA-E7 房间和 RMA 房间皮下地分别地在 syngeneic 老鼠被接种，肿瘤尺寸被一个星期，和 E7 蛋白质表示在老鼠的肿瘤织物滑动 caliper 两次测量被西方的污点在肿瘤以后检测形成。在忍受肿瘤的老鼠的 E7 特定的 T 房间的 cytolytic 活动的动力学被 LDH 工具包测量。再结合向量定序的结果证明被插入到 recombinant 的目标基因是正确的，并且稳定地表示 E7 蛋白质的 RMA-E7 房间被有限冲淡试金获得。在 vitro 在在 RMA 细胞和 RMA-E7 细胞之间的 morphous 和生长速度没有明显的差别。RMA-E7 房间在 syngeneic 老鼠成长了比 RMA 房间显著地慢。E7 蛋白质被表示在在 vivo 的 RMA-E7 房间更强壮比在 vitro。E7 特定的 CTL 的 cytolytic 能力在早阶段被激活，在中间的阶段到达了最大值，并且在结束阶段输了。从忍受肿瘤的老鼠孤立的 RMA-E7 房间对比在 vitro 有教养的 RMA-E7 房间杀死的 E7 特定的 CTL 更抵抗。E7 蛋白质表示没在 vitro 在 RMA-E7 房间的生长上有明显的影响的结论，和罐头在 vivo 压制 RMA-E7 房间的生长。E7 特定的 CTL 的活动曲线近似介绍鈥渂e ll 鈥 ? 形状。RMA-E7 房间在有的 vivo 成长了高表情 E7 铺平蛋白质，和对比在 vitro 有教养的那些杀死的 E7 特定的 CTL 抵抗的更多。在 vivo 的 E7 蛋白质表示不仅开始有免疫力的激活，而

CTL*

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Identification of two-dimensional electrophoresis-separated proteins in human hepatoma cell by electrospray ion trap mass spectrometry

中国科学通报：英文版 (2000)

Rong Zeng Lirong Yu Xiaoxia

As one of the most important analytical methods in proteome research, mass spectrometry was utilized to identify proteins separated by two-dimensional electrophoresis in the human hepatoma cell line BEL-7404. The protein spots were excised from the gel, followed by in-gel digestion, and the peptide mappings were analyzed by liquid chromatography electrospray ion trap mass spectrometer. Nine proteins were identified via database searching, according to the molecular weights and amino acid sequences of peptides, among which two proteins have not been identified in the other liver-cell database. The sequence coverage was 21% -72%. Furthermore, the relationship between the expressed proteins and the liver carcinoma was discussed.

Bottom-up proteomics

Proteome

Top-down proteomics

Peptide mass fingerprinting

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Differences in the primary culture, purification and biological characteristics between endothelial cells and smooth muscle cells from rat aorta

南京医科大学学报：英文版 (2009)

Shaobo Hu Zifang Сонг Qichang

目的将从老鼠主动脉调查在 endothelial 房间和光滑的肌肉房间之间的主要文化，纯化和生物特征的差别。方法 Endothelial 房间用脉管的戒指坚持， collagenase 消化方法和一个改进脉管的戒指坚持方法被获得，当光滑的肌肉房间与老鼠主动脉的织物节被分开时。endothelial 房间的克隆被限制冲淡试金选择。两种房间类型用特定的房间免疫被识别荧光灯的标记，和阶段对比显微镜学被用来在单个房间和殖民地水平观察在 endothelial 房间和光滑的肌肉房间之间的词法不同。房间增长被数 kit-8 的房间决定。在 cryopreservation.Results Endothelial 房间被所有三个方法获得以后， endothelial 房间和光滑的肌肉房间之间的差别在胰岛素消化时间，附件时间和恢复被评估。改进脉管的戒指方法提供了最可再现的结果。房间处于好状况，并且高纯净。光滑的肌肉房间由组织碎片文化方法成功地是有教养的。单个 endothelial 房间的同种细胞的扩大被达到。二种房间类型表示了他们的各自的特定的标记，并且光滑的肌肉房间的增长的率超过了 endothelial 房间的。Endothelial 房间比光滑的肌肉房间对胰岛素消化更敏感。另外，他们比光滑的肌肉房间有一更短的坚持时间和更好的恢复追随者 cryopreservation。结论改进脉管的戒指方法为产出 endothelial 房间是最佳的。限制冲淡是为从老鼠主动脉净化主要 endothelial 房间的一个新奇、有效的方法。主要老鼠 endothelial 房间和脉管的光滑的肌肉房间文化展出了不同词法特征，增长率，坚持时间，危险性到在 cryopreservation 以后的胰岛素消化和恢复。我们的研究能是为在血容器的新生的进一步的申请的一个好基础。

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