ABSTRACT
Objective To establish a stable HCV full-length genome replication cell model which is labeled with reporter gene and easyly to quantify intracellular HCV proteins and RNA level. Methodsneo gene was inserted into Luc-JC1 to make Luc-JC construct. Luc-JC RNA was obtained by in vitro transcription and then delivered into Huh7 cells by transfection. G418-resistant clones of Huh7 cells were obtained by selection. Clones of HCV full-length genome replication cell were confirmed by luciferase activity assay, Western blot and cleaveage of eYFP-MAVS by HCV NS3/4A protease. Then, HCV replication cell colonies were treated by different dose IFN-α in order to observe the change of luciferase activity, HCV protein and RNA level. Results At 3-4 weeks post-transfection, visible colonies were selected and stained by crystal violet. Luciferase activity and HCV NS3, NS5A protein were detected by luciferase activity assay and Western blot, respectively. Subcellular localization of eYFP-MAVS transferred from mitochondria to cytoplasms by cleavage of NS3/4A protease in cell colonies. Luciferase activity, HCV protein and RNA diminished obviously after IFN-α treatment. Conclusion A stable HCV full-length genome replication cell model labeled by reporter gene was successfully established and reporter activity can be used to indicate level of HCV proteins and RNA in cells. This cell model is a useful tool for the study on HCV pathogenesis and the screening of antiviral drugs.
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Objective To determine the value and limitation of water enema spiral CT (WESCT) in staging of colorectal carcinoma.Methods Forty-eight patients with histologically proven rectum or colon carcinoma were included in this study. All of them were examined by SCT ,and the preoperative staging of TNM and Duke were used based on the findings of SCT.The results of WESCT were compared with those of surgical and pathological examination in all cases. Results All lesions in the 47 cases were demonstrated clearly by WESCT and the sensitivity was 97.9% ; 39 cases of 48 patients were correctly staged with TNM and 42 cases with Duke , the accuracy was 81.3% and 87.5% respectively, which were higher than the overall 50 % accuracy reported by references ;⑶The accuracy of WESCT was 89.6% (43/48) in T stage and 81.3% (39/48) in N stage. Three casas in M stage were all diagnosed correctly;Conclusion WESCT scan is a better method of depicting the colorectal carcinoma. It allows for accurate depiction and staging of colorectal carcinoma, especially detecting the invasion of adjacent tissues and distant metastasis. It is the best imaging method for staging the colorectal carcinoma . However the value of WESCT for early T staging in colorectal carcinoma and minute metastasis of lymph nodes or liver is limited.
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Objective To evaluate the diagnostic value of TNM staging of gastric carcinoma with triphase incremental helical CT.Methods Triphase incremental helical CT of the stomach with water-filling method were performed in 101 patients with gastric carcinoma.Results (1)The accuracy of T-staging,N-staging and TNM staging was 81.8%,72.9% and 80.2% respectively.(2)Demonstrating multilayered structure of the normal gastric wall in the arterial parenchymal-phase was advantageous to improve the accuracy of estimating the depth of tumor invasion (?
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Objective To evaluate the diagnostic value and clinical significance of preoperative staging in gastric cancer with induced pneumoperitoneum in spiral CT (SCTPP). Methods Both routine SCT and SCTPP were performed in 52 lean patients suffered from gastric cancers, and comparison was made between SCT findings and surgical and histopathologic findings. Results The accuracy of routine SCT and SCTPP in determining the T-staging was 72% and 96%, respectively (?2=8.0, P0.05). The sensitivity in determining M-staging was 61% and 100%, respectively (?2=0.04, P