Construction of eukaryotic expression plasmid pEGFP-N3-M-IL-2(88 Arg,125 Ala) and its effects on Glioma cell line U87 / 中国免疫学杂志

Objective:To construct a recombinant eukaryotic expressive vector of pEGFP-N3-M-IL-2(88 Arg,125 Ala),and to study the expression of this gene in the Glioma cell line U 87,and to detect its antitumor activities of the fusion protein M-IL-2(88 Arg,125 Ala).Methods:The target fusion gene M-IL-2 (88 Arg,125 Ala) was amplified by PCR from pPICZαA/M-IL-2 (88 Arg,125 Ala) and cloned into pEGFP-N3 vector after digestion to construct recombinant eukaryotic expressive vector pEGFP -N3-M-IL-2( 88 Arg,125 Ala).And then recombinant plasmid pEGFP-N3-M-IL-2(88Arg,125Ala) was transfected into Glioma cell U87 by LipofectamineTM2000 immediately after it was confirmed by restrictive enzyme analysis and sequencing .RT-PCR and Western blot were used to confirm expression of the fusion gene in the U87.Prohibitory effect of recombinant M-IL-2(88Arg,125Ala) protein on U87 was assessed by CCK-8 assay.Results:Restrictive analysis and sequence analysis revealed that M-IL-2(88Arg,125Ala) fusion gene was cloned into the vector pEGFP-N3 suc-cessfully,fusion gene M-IL-2(88Arg,125Ala) could express in U87 cells and could inhibit the growth of U87 cells.Conclusion:The eu-karyotic expression plasmid pEGFP-N3-M-IL-2( 88 Arg,125 Ala) was constructed and expressed in U 87 cells successfully ,the fusion protein could inhibit the growth of U87 cells.We laid a foundation for further research of gene M-IL-2(88 Arg,125 Ala).

Expression of NF-κB, TNF-α and IL-6 in colorectal carcinoma and their clinical significances / 肿瘤研究与临床

Objective To investigate the expression of nuclear factor kappa B (NF-κB),tumor necrosis factor α (TNF-α) and interleukin6 (IL-6) in human colorectal carcinoma tissues,and to explore their clinical significances in the genesis and development of colorectal cancer.Methods Sixty cases of colorectal cancer tissues and 36 cases of colorectal adenoma tissues were collected,60 cases of paracancerous normal colorectal tissues were the controls.Immunohistochemistry SABC method was used to detect the expression of NF-κB,TNF-α and IL-6 in each group respectively.The correlation of NF-κB,TNF-α and IL-6 with clinical pathologic features of colorectal cancer was analyzed.Results In colorectal carcinoma,adjacent normal colorectal tissues and colorectal adenoma tissues the positive expression rates of NF-κB were 76.7 ％ (46/60),46.7 ％ (28/60),83.3 ％ (30/36),the positive rates of TNF-α were 70.0 ％ (42/60),36.7 ％ (22/60),66.7 ％(24/36),the positive rates of IL-6 were 80.0 ％ (48/60),43.3 ％ (26/60),61.1％ (22/36).The differences were significant in each group (all P ＜ 0.05).The expression of NF-κB was closely associated with the expression of TNF-α and IL-6 respectively.In addition,the expression of NF-κB and TNF-α were correlated with vascular invaded,lymphnode metastasis and different stages.The expression of IL-6 was correlated with lymphnode metastasis and different stages.Conclusion The over expression of NF-κB and the downriver inflammation factors have close relationship with biological behaviors of colorectal cancer.It may be considered that the pathway of NF-κB play an important role in the genesis and development of colorectal cancer.

Inhibitory Effects of Ginsenoside Rb1 on Apoptosis Caused by HSV-1 in Human Glioma Cells / 中国病毒学·英文版

To investigate the inhibitory effects of Ginsenoside Rb1 (GRb1) on apoptosis caused by Herpes Simplex Virus-1 (HSV-1) in Human Glioma Cells (U251),U251 cells were infected by HSV-1 at a multiplicity of infection of 5 and GRb1,GRb1+HSV-1,HSV-1 and control groups.MTT and cell apoptosis assays were used to detect the inhibitory effects of GRbl on the apoptosis of U251 cells that caused by HSV-1 infection for various concentrations of drug and virus treatments by MTT assay.We found that in the 400 μg/mL GRbl and 400 μg/mL GRbl+HSV-1 groups,MTT values were higher than control group at all times (P＜0.05).Moreover,the apoptosis rate in the 400 μg/mL GRb1+HSV-1 group was lower than the HSV-1 group (P＜0.05).These results confirmed that,at appropriate concentrations,GRb 1 could inhibit nerve cell apoptosis in HSV-1 infections.

HCMV Infection Depress NGF Expression in Human Glioma Cells / 中国病毒学·英文版

Human cytomegalovirus (HCMV) is the most common cause of congenital infection, resulting in birth defects such as microcephaly. In this study, RT-PCR and Western Blotting were performed to quantify the regulation of endogenic nerve growth factor expression in neuroglia cells by HCMV infection. The results showed that basal, endogenous NGF expression in U251 was unchanged during early HCMV infection. NGF expression is strongly down-regulated during the latent phase of infection. These results suggest that HCMV can depress the NGF expression in U251 cells.

Gene expression differential study in peripheral blood mononuclear cell(PBMC) of patients with SLE and IDDM / 中国免疫学杂志

6).The genes were genes that associated with the regulation of cell differentiation and proliferation,adhesion and signal tranduction,apoptosis,transcription and modulation,and DNA damage and repair that.Conclusion:The differential hybridzation analysis of Atlas cDNA expression arrays can be a useful method for analysing the expression profiles of PBMC genes and studing genes differential expression in SLE and IDDM patients.