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To establish a cell-based rapid luciferase suppression assay for high-throughput screening (HTS) anti-alphaviruses compounds screening, which could cause viral encephalitis, raise the social issues associated directly with public health and huge economic burden to the society. The Gaussia luciferase assay system was used for HTS model for identifying inhibitors of labeled virus XJ160-GLUC. The decreased 50% GLUC activity inhibition ratio was deemed to be the screening positive index. The reaction system in this model was optimized, and the reliability of the model was evaluated. For HTS model's optimization, cells were infected with XJ160-GLUC at an MOI of 0.025 PFU/cell. The supernatant treated with compounds 48h were collected for GLUC expression detection. In the model, Z' factor was up to 0.71, demonstrating that HTS assay for identifying inhibitors that target all aspects of the viral life cycle of XJ160-GLUC was stable and reliable. After screening 8080 compounds (five-in-one), 341 positive samples were selected, and the positive rate was 4.2% with a cutoff at 50% inhibition. Then 1705 compounds were screened subsequently and the positive rate was 1.1% with obtaining 19 positive compounds. These results will lay the foundation for finding the anti-alphaviruses' drug targets.
Alphavirus ; drug effects ; genetics ; metabolism ; Animals ; Antiviral Agents ; pharmacology ; Drug Evaluation, Preclinical ; methods ; Genes, Reporter ; High-Throughput Screening Assays ; methods ; Luciferases ; genetics ; metabolism

9.The quality analysis of the merged culture of the training for professional master degree students of clinical medicine and resident standardization training in Zhejiang Province

Weiji YANG ; Jianhua PENG ; Xianfang LUO ; Chenzi WUYANG ; Weijun ZHENG ; Jiayang QI ; Bin FU

Chinese Journal of Medical Education Research 2017;16(4):330-333

Objective To improve the training system of the combined education mode between professional master degree students of clinical medicine and resident standardization training (RST) in Zhe-jiang province, and improve the professional quality of medical graduate students. Methods A question-naire was designed through literature review and expert interview, and the data of 77 clinical training post-graduates were collected, and the effect of the training was evaluated by taking the reaction level, learning level, behavior level and achievement level of Ke's evaluation as the breakthrough point. Statistical analysis was performed by SPSS 20.0, and the mean score was used as the standard deviation. The scores were compared with t test, and the scores of multiple groups were made variance analysis. Results The RST attitude value in reaction layer was 9.26±1.08, RST content value was 29.29±4.36, RST teacher enthusiasm value was 11.13±2.17, RST experience value was 17.38±3.10, which indicated that residents' overall satisfaction to RST was high, but the satisfaction on teachers, evaluation and compensation was relatively low;The grade difference before and after RST in learning layer was statistically significant (P<0.05), indicating that before and after RST the grades were obviously improved, and the sense of belonging is high. The behave layer showed the residents' behavior changed obviously before and after RST (P<0.05), and their abilities in various aspects such as ward round and dealing with common diseases were largely improved;The result layer showed 77 graduate students had no medical accident and complaint , and all of them passed the annual assessment and participated in the research projects. Conclusion In general, RST graduate students are relatively satisfied with the plan, content and methods of the training, on the other hand, there are problems such as the absence of timely information feedback, teachers' poor enthusiasm, lack of effective competition mechanism, etc. Kirkpatrick Model made a comprehensive and objective eval-uation on graduate students RST from a overall perspective, and it can be introduced into the effectiveness evaluation for graduate students RST.