Molecular biology is the fundamental course of life science,and its experimental teaching is difficult to offer due to its long process and need of expensive equipments.Virtual laboratory platform is an important approach for practice teaching in recent years.This article discussed the advantages and requirements,provided the experience of its applying at our university,and suggested that virtual laboratory platform can play a key role in molecular biology experimental teaching.

Objective To explore the effect of the gene silencing of phosphatidic acid-preferring phospholipase A1 (PA-PLA1) on insulin secretion in mouse insulin-secreting cell line MIN6. Methods The siRNA expression vector of mouse PA-PLA1 gene targeting was constructed using mouse PA-PLA1 mRNA sequence available in GenBank, and MIN6 cells were transfected with the vector. Fluorescence quantitative PCR and Western-blotwere applied to screen efficient RNAi-vector. After transfection with obtained efficient RNAi-vectors for 48 hours, glucose-stimulated insulin secretion experiments were conducted, and the changes of insulin secretion were examined. Results Four siRNA expression vectors of mouse PA-PLA1 gene targeting were confirmed to be successfully constructed by the analyses of enzyme cleavage and sequencing. The results of fluorescence quantitative PCR and Western blot analyses indicated that the siRNA expression vectorpGPU6-PA-PLA1-1885was the most effective RNAi-vector in the four vectors. The expression levels of the PA-PLA1 mRNA and protein of the MIN6 cells transfectedwith pGPU6-PA-PLA1-1885 decreased to 46.3% and 33.9% of that of the control, respectively, and meanwhile the insulin secretion levels of the cells decreased to 65.0% of that of the control (P < 0.05). Conclusion The gene silencing of phosphatidic acid-preferring phospholipase A1 might decrease insulin secretion in MIN6 cells.

Objective To explore the effect of hepatitis B virus (HBV) X protein (HBx) on autotaxin (ATX) expression and its significance. Methods The recombinant eukaryotic expression vector of HBx ,pcD-NA3.1(+)-HBx,and the recombinant luciferase reporter gene vector of ATX promoter,pGL3-ATX,were con-structed and used to co-transfect HepG2 cells to examine the effect of HBx on the activity of ATX promoter. The sta-ble cell expressing HBx,HepG2.HBx,was constructed,and Western blot(WB)was used to detect the effect of HBx on ATX expression. Results The luciferase activity of pcDNA3.1(+)-HBx and pGL3-ATX group was 1.47 times as that of the empty vector cDNA3.1(+)and pGL3-ATX group(P<0.000). WB detection showed that the expression of ATX protein was increased in HepG2.HBx cells,and 1.75 times as that of HepG2 cells(P<0.05). Conclusion HBx can activate ATX promoter and up-regulate ATX expression ,thus suggests that HBV infection might enhance ATX/LPA signaling.

It remains a common problem that the evaluation model of course learning for stu-dents is so simple in the higher education of China. Moreover, the teachers are usually used to evalu-ating student's course learning by summative appraisal and ignore process appraisal, so that the stu-dents are usually lack of learning enthusiasm for self study. Besides, the course learning evaluation of students generally lacks the uniform and detail standard in the country. To resolve these problems, a comprehensive evaluation model of course learning based on the principle of talent evaluation has been practiced and improved since 2007 in some basic medical courses at our university. This model is composed of four parts:the evaluation of mastering basic knowledge, the evaluation of the ability to resolve problems through applied knowledge, the evaluation of practical study and the evaluation of learning initiatives. The model has showed in the practice the characteristics of powerful operability, pluralistic evaluating indicators, combining summative appraisal and process one, reflecting objectively the mastering degree of expertise of students and promoting the self-study of students.

Objective:To screen the interacting protein of ubiquitin-conjugating enzyme E2S(UBE2S)and construct the hepatocellular carcinoma(HCC)based on UBE2S interacting protein prognosis model(UIPM),and to discuss the value of UIPM in assessing the prognosis of the HCC patients.Methods:Co-immunoprecipitation(Co-IP)was used to screen the protein complexes binding to Flag-UBE2S.After validation by sodium dodecyl sulphate-polyacrylamide gel electrophoresis(SDS-PAGE)and Western blotting methods;liquid chromatography-mass spectrometer(LC-MS)was used to identify the UBE2S interacting proteins;Gene Ontology(GO)functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)signaling pathway enrichment analysis were conducted on these proteins;the prognosis-related proteins from The Cancer Genome Atlas(TCGA)were cross-referenced with UBE2S interacting proteins by survival package of R software;the key proteins were extracted through LASSO regression analysis to build the UIPM;the prognostic model risk scoring formula was established.The HCC patients in TCGA were divided into high risk group and low risk group based on median value of the risk scores.The predictive accuracy of UIPM was evaluated by receiver operating characteristic curve(ROC),and the predictive accuracy was further validated by International Cancer Genome Consortium(ICGC)Database;univariate regression analysis and multivariate Cox regression analysis were used to detect whether the UIPM risk score was an independent prognostic factor for HCC.Furthermore,the nomogram model was built.Results:A total of 97 UBE2S interacting proteins were identified through Co-IP combined with LC-MS analysis.The GO functional enrichment analysis and KEGG signaling pathway enrichment analysis results showed that the interacting proteins were closely associated with cysteine-type endopeptidase activity,oxidative stress,and cell death.The TCGA revealed 5 163 HCC prognosis-related proteins;after intersecting with UBE2S interacting proteins,40 prognosis-related interacting proteins were found.Seven key proteins were determined through LASSO regression analysis,including UBE2S,heat shock protein family A member 8(HSPA8),heterogeneous nuclear ribonucleoprotein H1(HNRNPH1),chaperonin containing TCP1 subunit 3(CCT3),eukaryotic translation initiation factor 2 subunit 1(EIF2S1),receptor for activated C kinase 1(RACK1),and actin related protein 2/3 complex subunit 4(ARPC4),and the UIPM was constructed.There was significant difference in survival rate of the patients between high risk group and low risk group(P<0.05).The ROC curve analysis results showed the area under ROC curve(AUC)values of UIPM for predicting 1-year,2-year,and 3-year survival risk scores of the HCC patients were all greater than 0.7,indicating the model had high predictive accuracy.This was also confirmed by ICGC Database data.The univariate and multivariate Cox regression analysis results showed that the UIPM risk score was an independent prognostic risk factor for the HCC patients(P<0.05).The nomogram results showed good consistency between predicted survival rate and actual survival rate of the patient.Conclusion:A total of 97 interacting proteins that interact with UBE2S may promote the occurence and devolopment of HCC through oxidative stress and dysregulation of ferroptosis pathways.The UIPM risk score is an independent risk factor for the prognosis of HCC and can be used to predict the outcomes of the patients.UBE2S,HSPA8,HNRNPH1,CCT3,EIF2S1,RACK1,and ARPC4 could be regarded as the new biomarkers and therapeutic targets for HCC.