Objective:To explore the perioperative dynamic changes of cellular immune function and its clinical significance in patients with chest surgery.Methods:The numbers of CD3、CD4、CD4/CD8、CD8 T lymphocyte and nature killer(NK) cells in peripheral blood were examined in 45 patients with chest surgery before operation and 1、3、5 and 7 days after operation by flow cytometry.45 patients were devided randomly into groups.The perioperative changes of T lymphocyte subsets and NK cells were compared among mediastinal disease,pulmonary operation,esophagus and gastric cardia surgery.Results:CD4/CD8 decreased 1 day after surgery and CD4 decreased at the 3rd postoperative day in patients with chest surgery.In patients with mediastinal disease,CD4/CD8 decreased 1 day after surgery and CD4 decreased at the 3rd postoperative day.In patients with pulmonary operation,CD8 decreased at the 1st day and 7th day after surgery.NK cells decreased at the 5th postoperative day CD3 increased at the 5th day after surgery.CD4/CD8 increased at the 7th day after operation.In patients with esophagus and gastric cardia surgery,CD4/CD8 decreased 1 day after operation,CD3 and CD4 increased at the 7th postoperative day.All differences are statistically significant( P

OBJECTIVETo investigate the roles of Domain I and Domain II of hepatitis C virus (HCV) 5' untranslated region (UTR) in the translation initiation activity of HCV 5'UTR in different host cell lines.

METHODSThe eukaryotic expression plasmid pCMVNCRLuc (pCN1), in which full-length HCV 5'UTR regulates firefly luciferase expression, was modified by deleting Domain I and the downstream single-stranded sequence (43 bp in total) from the UTR (pCNl-d2), Domain I with the downstream single-stranded sequence and Domain II (118 bp in total) from the UTR (pCNl-d3), or the total UTR (pCNl-d5). The modified plasmids were transfected via liposome into different cell lines with pRL-TK plasmid co-transfected as the normalization control. At 36 h after the transfection, the total cellular RNA was harvested for semi-quantitative RT-PCR, and the relative expression activities of luciferase were assayed with a dual luciferase reporter gene assay system. The translation initiation activities of the truncated HCV 5'UTRs in different translation systems were analyzed.

RESULTSDeletion of Domain I and the downstream single-stranded sequence caused no significant changes of the translational activity of HCV 5'UTR in Hela or C6 cells, but decreased the translational activity by 46% in L-02 cells and increased the translational activity by 46% in 293T cells. Deletion of both Domain I and Domain II resulted in decreased translational activity of HCV 5'UTR by 51% in HeLa cells, but increased the translational activity by 40% in L-02 cells, 60% in C6 cells and 135% in 293T cells.

CONCLUSIONSDomain I and Domain II of HCV 5'UTR perform cell type-specific roles in HCV IRES-driven translation initiation.

5' Untranslated Regions ; Genes, Reporter ; HeLa Cells ; Hepacivirus ; genetics ; Humans ; Luciferases ; Plasmids ; Protein Biosynthesis ; genetics ; RNA, Viral ; genetics ; Transfection