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| 1 | 植物凝集素刺激外周血单个核细胞增殖及分泌因子表达的变化显示文摘背景:植物凝集素可以刺激外周血单个核细胞分裂并引起免疫细胞的活化,是常用的细胞增殖模型。但是全血中的非外周血单个核细胞成分(血浆和无核细胞)在植物凝集素参与的体外培养过程中起到什么作用,以及内皮分泌因子的表达情况尚不明确。目的:在体外单独培养或全血培养外周血单个核细胞过程中,观察植物凝集素对其增殖和凋亡的影响,以及相关炎症因子以及内皮分泌因子的表达变化。方法:分离正常核型成人的外周血单个核细胞,应用无植物凝集素培养基和加入植物凝集素培养基分别进行单独培养,观察细胞形态学变化。收集全血培养或不同条件单独培养的外周血单个核细胞,提取mRNA,荧光定量RT-PCR检测细胞增殖、凋亡,以及炎症因子和内皮分泌因子的表达变化,并进行统计学分析。结果与结论:外周血单个核细胞单独培养和全血培养存在差异,植物凝集素会促进外周血单个核细胞Ki67、增殖细胞核抗原、蛋白水解酶3、γ-干扰素、肿瘤坏死因子β和白细胞介素6的基因表达,抑制蛋白C表达。提示植物凝集素促进体外培养外周血单个核细胞的增殖及凋亡,促使炎症因子表达上调,部分血液生物学相关的内皮分泌因子下调。 | 王鼎 宋兵 钟旋 孙筱放 范勇 | 2014 | 中国组织工程研究2014，18，23： | 4 |
| 2 | Preparation and Identification of scFv and bsFv against Transferrin Receptor显示文摘To obtain single chain variable fragment (scFv) and bivalent single chain variable fragment (bsFv) against transferrin receptor, up-stream and down-stream primers were designed according to the complementary sequences of FR1 region of variable heavy (VH) and FR4 of variable light (VL), respectively, which contained inter-linker G4S and the restriction endonuclease SfiI, AscI and NotI. Two pieces of scFv fragments were first amplified through PCR and then inserted into plasmid pAB1, which could express scFv protein once induced by IPTG in the host bacteria. To express scFv and bsFv, E. coli TG1 was cultured in LB broth and was induced by IPTG. The restriction enzyme digestion map and DNA sequencing demonstrated that scFv and bsFv genes were successfully inserted into the expression plasmid. SDS-PAGE and Western blotting revealed the protein band at 35kD and 60kD, which were consistent with the molecular weight of scFv and bsFv respectively. Flow cytometry showed that scFv and bsFv harbored the specific binding activity with TfR expressed in various tumor cells, and the avidity of bsFv was higher than that of the parent scFv. | 刘静 肖代雯 周小鸥 文雪 代红 王志华 沈昕 代维 杨道锋 沈关心 | 2008 | Journal of Huazhong University of Science and Technology(Medical Sciences)2008，28，6： | 0 |
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