Preliminary practice of the resident physicians in specialized hospital participating in the stan-dardized clinical resident training in general hospital / 中华医学教育探索杂志

The significance for specialized hospital's resident physicians participating in the standardized clinical resident training in general hospital is expounded and its existing prob-lems are pointed out.It is suggested that the administration division should give full play to subjective initiative,coordinate problem solving and improve training quality,and meanwhile speciality hospitals should make an active effort in the application for national subspeciality physician training base.

Dilemma and strategy of clinical teaching of infectious disease in non-affiliated hospitals of university / 中华医学教育探索杂志

With more and more non-affiliated hospitals of universities joining the clinical teaching of infectious diseases,there have been some dilemmas on clinical teaching,such as the shortage of staff,teaching experience,poor lecture art and teaching enthusiasm,textbook lag,the shortage of case teaching abd students'fear of infectious disease etc. Countermeasures such as perfecting the organizational structure,formulating preferential policy,clearing historical mission,increasing students' interest in learning,adding new academic progress,enriching teaching methods,setting up experts'supervision will ensure the effective teaching quality.

Screening and cloning gene of hepatocyte protein interacting with hepatitis C virus core protein / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To clone the unknown gene of hepatocyte protein interacting with hepatitis C virus core protein.</p><p><b>METHODS</b>Using the yeast dual hybrid system 3, bait plasmids of hepatitis C virus core were constructed. After identifying hepatitis C virus core protein that could stably expressed in AH109 yeast strains, we performed yeast two hybrid by mating AH109 with Y187 that transformed with liver cDNA library plasmids pACT2 and then plated on quadrople dropout (QDO) medium and assayed for alpha-gal activity. The genes of yeast colonies that could grow on QDO and had alpha-gal activity were sequenced.</p><p><b>RESULTS</b>Among the 30 positive colonies, we blasted the gene of the sixth colony; we coined human hepatitis C virus binding protein 6(Hu Hcbp6) with Genbank, realized that the Hu Hcbp6 shares as much as 98% homology with two cDNA without knowing functions. We have proved that Hu Hcbp6 could interact with hepatitis C virus core protein.</p><p><b>CONCLUSIONS</b>Hepatitis C virus core binding protein (Hu Hcbp 6 Genbank number: AY032594) was successfully cloned and identified. The study partly paved the way for investigating physiological function of the Hu Hcbp6.</p>

Synergetic transactivating functions of HCV core and truncated HBV middle surface protein / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the synergetic transactivating functions of HCV core and truncated HBV middle surface proteins.</p><p><b>METHODS</b>Two recombinant expression plasmids harboring HCV core and C-terminally truncated HBV middle surface protein gene were constructed, respectively. The plasmids were transfected into HepG2 cells and cotransfected HepG2 cells with reporter plasmid pSV-lacZ by lipofectamine plus reagents. The transient expressed viral proteins were identified at the transcription and translation levels. The activity of beta-galactosidase was detected, which reflected the transactivating function of the proteins.</p><p><b>RESULTS</b>The protein expression of plasmids was detected in soluble cell extracts of transiently transfected HepG2 cells. HCV core protein activated the beta-galactosidase expression at a value of 4.6 times higher than the control, while C-terminally truncated HBV middle surface protein activated at a value of 3.2 times. It reached 8.4 times transfected with the plasmids simultaneously. The transactivating effect was dose dependent.</p><p><b>CONCLUSIONS</b>It is suggested that the two kinds of virus proteins have transactivating effect on SV40 early promoter/enhancer, and they act synergistically. These contribute to explain the mechanisms of liver injury or tumorigenesis induced by HCV or/and HBV infection.</p>

CLONING AND EXPRESSION OF SOLUBLE HCV NS5B GENE IN EUKARYOCYTES / 解放军医学杂志

HCV NS5B, acting as a RNA dependent replication enzyme,has emerged as an attractive protein used as a target for screenig of drugs against HCV NS5B, and plays an important role in HCV replication. In the report the gene expression of NS5B in E.coli was investigated. PCR was performed to gain the gene of HCV NS5B from plasmid pBRTM/HCV 1 which contains whole sequence of HCV, and the truncated NS5B gene containing no hydrophobic domain was cloned into pGEM Teasy vector. The gene of the truncated NS5B was cut from pGEM Teasy vector and cloned into E.coli expression plasmid pET 21b, then pET 21b NS5B was transfected into E.coli cells. The protein E.coli lysates were purified and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) and Western blotting assay. The RdRp activity of NS5B was examined by scintillation proximity assay (SPA). The truncated NS5B gene was successfully cloned into pET 21b. The results of SDS PAGE and Western blotting assay showed: ①the molecular weight of the expressed product was about 68 000 D, ②The truncated NS5B protein was existed in media of E.coli cells, ③The activity of NS5BDCT21 His from HCV 1b amounted to 6 900 cpm (total incorporation of approximately). These findings suggest that soluble NS5B can be successfully expressed in E.coli and could secret into media.

ANGIOGENESIS OF VASCULAR ENDOTHELIAL GROWTH FACTOR EVALUATED USING RAT SPONGE IMPLANT MODEL / 解放军医学杂志

To construct a repetitive, objective and consecutive system for evaluating the angiogenesis of vascular endothelial growth factor(VEGF) in vivo, a sterile polyvinyl alcohol sponge disc was implanted into the subcutaneous part of rat and five days later pCMV 4 VEGF 165 expressed plasmid and empty vector pCMV 4 were injected into the sponge, respectively. 7 and 11 days after injection, the tissue ingrowth in the implants and expression of VEGF gene in the serum, the tissues in the sponge and the liver were measured by RT polymerase chain reaction (RT PCR) .The pathological section of sponge was stained with hematoxylin and eosin (HE) and immunohistochemistry of Ⅷ factor specific for vascular endothelial cells. The tissue ingrowth in the implants was analysed by image analysis system and statistical analysis was done with Stata Software. The results indicated that the expression of VEGF gene was located at the injection site and promoted the tissue ingrowth in the implants. In brief, the angiogenesis of VEGF was successfully observed using the sponge implant model, which provides lays the theoretical foundation for local gene therapy of ischemic diseases.