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Objective:To obtain highly purified recombinant human IGF-Ⅰ(rhIGF-Ⅰ) and identify it.Methods:rhIGF-Ⅰ Was purified through ion-exchange chromatography and gel filtration chromatography after the inclusion bodies of rhIGF-Ⅰ were extracted from Escherichia coli. The recombinant protein was characterized through molecular weight assay, Western-blot, and fluorescent chromatography. The renaturation and biological assay of rhIGF-Ⅰ were investigated. Results and Conclusions: The purity of rhIGF-Ⅰ was higher than 99%. The analysis of molecular weight, Western-blot, fluorescent chromatography and sequences of NH2-terminal 15 amino acids were same as those anticipated. 3-10 mg/ml was the concentration of renatured rhIGF-Ⅰ to support half-maximal stimulation of cell proliferation with BALB/c 3T3 cells.
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Purpose:To evaluate the antitumor effects of human cytokine-induced killer (CIK) cells from healthy donors and patients with primary hepatocellular carcinoma (HCC) in vitro and in vivo .Methods:Peripheral blood mononuclear cells (PBMC) from healthy donors and patients were induced to become CIK cells by in vitro incubation. The CIK cells were identified at different intervals by flow cytometry analysis. The cytotoxicity of CIK cells against autologous primary HCC was determined by 51Cr Release Assays. The antitumor activity of the CIK, LAK and PBMC cells were evaluated in Balb/c nude mice bearing BEL-7402liver cancer.Results:The flow cytometry analysis showed that CIK cells from patients with HCC proliferated and expanded more than 300-fold in vitro incubation by day 28;the percentage of CD3+CD56+ cells increased from 0.23% on day 0 to 17.8% on day 21. CIK cells generated from patients with HCC patients possessed a higher antitumor cytotoxic activity on autologous HCC cells in vitro than autologous PBMC. In addition, CIK cells had a stronger suppressive effect on the tumor growth in Balb/c nude mice bearing BEL-7402 tumor than LAK cells (median inhibitory rates 84.7% vs 52.8%, P