ROG,GATA3 and T-bet mRNA levels in peripheral blood mononuclear cells in patients with chronic hepatitis B / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the role of ROG, GATA3 and T-bet in the progression of chronic hepatitis B (CHB).</p><p><b>METHODS</b>The mRNA levels of ROG, GATA3 and T-bet in peripheral blood mononuclear cells (PBMCs) from 135 patients with CHB (including 45 mild cases, 42 moderate cases, and 48 severe cases) and 15 healthy control subjects were detected by real-time quantitative PCR.</p><p><b>RESULTS</b>The levels of T-bet mRNA in the PBMCs were significantly higher in CHB patients than in the healthy controls (P<0.05), and also differed significantly between the 3 groups of CHB patients (P<0.05). ROG mRNA levels were significantly higher in severe cases of CHB than in the healthy controls and mild and moderate CHB cases (P<0.05), but were similar among the latter 3 groups (P>0.05). The mRNA level of GATA3 in the PBMCs were significantly higher in moderate and severe CHB cases than in the healthy controls and mild CHB cases (P<0.05). The T-bet/GATA3 ratio was significantly greater in the 3 CHB groups than in the control group (P<0.05) but comparable between the 3 CHB groups (P>0.05). ROG levels were not correlated with GATA3 levels or T-bet/GATA3 ratio in the CHB cases.</p><p><b>CONCLUSIONS</b>The mRNA levels of ROG, GATA3 and T-bet in the PBMCs are obviously up-regulated in CHB patients and these 3 genes may participate in the progression of CHB. ROG plays an important role in correcting and maintaining the new balance of Th1/Th2.</p>

Construction and expression of non-structural protein gene 3-4b of HCV 1b based on the adenoassociated virus vector / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To clone 1b type of HCV NS3-4b Gene and express in HEK 293 cells, lay the foundation for further study of the HCV NS3-4b recombinant adeno-associated virus vaccine and its dendritic cell vaccine.</p><p><b>METHODS</b>HCV 1b patients' serum was collected, and full length NS3-4b segment was amplified by RT-PCR and cloned into adeno-associated virus' expression vector pAAV. CMV. EGFP in order to express in HEK 293 cells. At last, it was validated whether express or not by Western Blot.</p><p><b>RESULTS</b>The 1b type gene NS3-4b were amplified and consistent to the expected size (2838 bp), the recombinant plasmid has been confirmed its successful restructured by double enzyme and sequencing, at last, Western Blot map can see objective protein expression after it transfect HEK 293 cells.</p><p><b>CONCLUSION</b>The adeno-associsted virus recombination HCV NS3-4b plasmid have successfully constructed and it can express in eukaryotic cells.</p>