Objective To evaluate liver protection provided by ulinastatin in rats with liver fibrosis.Methods Fifty pathogen-free male Sprague-Dawley rats,weighing 180-220 g,aged 6-8 weeks,were randomly divided into 5 groups (n =10 each) using a random number table:control group (group C),carbon tetrachloride (CCl4) group,low-dose ulinastatin group (group L),medium-dose ulinastatin group (group M),and high-dose ulinastatin group (group H).Hepatic fibrosis was produced by subcutaneous injection of 50％ CCl4 peanut oil solution two times a week for 8 weeks.After hepatic fibrosis was produced (at 9th week),ulinastatin 2.5× 104,5.0× 104 and 10.0× 104 U/kg were injected via the caudal vein in L,M and H groups,respectively,once a day for 7 days.Blood samples were collected after 24 h of fast on 8th day for determination of serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) concentrations by ELISA.The rats were then sacrificed and livers were removed for microscopic examination of pathologic changes with light microscope.The expression of interleukin-1β (IL-1β),IL-2,IL-6,IL-8,Toll-like receptor 4 (TLR4),and tumor necrosis factor-alpha (TNF-α) mRNA and protein was detected by RT-PCR and Western blot.Results Compared with group C,the serum AST and ALT concentrations were significantly increased in CCl4,L and M groups,the expression of IL-1β,IL-2,IL-6,IL-8,TLR4 and TNF-α mRNA and protein was up-regulated in CCl4 group,and no significant change was found in the parameters mentioned above in group H.Compared with group CCl4,the serum AST and ALT concentrations were significantly decreased in M and H groups,no significant change was found in the serum AST and ALT conccntrations in group L,and the expression of IL-1β,IL-2,IL-6,IL-8,TLR4 and TNF-α mRNA and protein was down-regulated in group H.The pathologic changes of hepatic tissues were attenuated in M and H groups as compared with group CCl4.The pathologic changes of hepatic tissues were almost recovered to the normal structure in group H.Conclusion Ulinastatin can produce liver protection in rats with liver fibrosis.

Objective Structured Clinical Examination(OSCE) is a model of clinical skills examination.We have applied OSCE in the graduate examination of medical colleges for three years.There are different requests for training teachers and Standardized Patients(SP),setting exam place,examiners and students and so on.OSCE is worth spreading as clinical ability examination in other medical education.

Objective To investigate the distribution of hepatitis B virus genotypes in Hangzhou area and preliminarily identify and evaluate the applied characteristics of oligonuleotides(oligo)microarray for genotyping hepatitis B virus(HBV).Methods HBV PCR products were hybridized with oligonucleotide probes,which were prestablized on the chip.The hybridized results were colorized.According to the hybridization signal and the corresponding probe sequence,HBV genotype was determined.Results Of the 106 HBV DNA positive patients 37(34.91%)were genotype B,and 69(65.09%)were genotype C,No genotype A,E and F were found in the studied subjects.Conclusions HBV genotype B and C exsited in Hangzhou No HBV genotype A,D,E and F were found in the studied subjects.We should take further investigation for HBV genotypes by enlarging study population.The advantages of this assay are sensitive accurate,fast and economical when using it for HBV genotype test comparing with other relative methods.

Objective To investigate the distribution and virologic characteristics of HBV genotypes,and possible association with the severity of liver disease.Methods HBV genotype was determined,using oligonuleotides(oligo) microarray method.Results 37patients were differentiated as genotype B,69 genotype C.The clinical manifestations demonstrated that the serum lever of HBVDNA and HBeAg positive rate in patients of genotype C was 7.02?1.26 and 60.87% higher than 5.62?1.02and 32.43% in those of genotype B,the serum HBeAb positive rate in patients of genotype B was 67.57% higher than 39.13% in those of genotype C.The occurrence rates of those developed to chronic hepatitis B liver cirrhosis and hepatoma in those of genotype C were 46.38%,30.44% and 11.59% highter than that of 40.54%,13.51%and 8.11% in those of genotype B.Patients with genotype B were much younger than those with genotype C.Conclusions Genotype B and C exsited in Hangzhou,genotype C is a predominated genotype.The serum leverl of HBVDNA in those of genotype C is highter than that of genotype B.The positive rate of HBeAg in those of genotype B is highter than that of genotype C.The damage to liver induced by genotype C is severe than that of genotype B.

AIM: Tumor necrosis factor-?(TNF-?) participates in the establishment of inflammatory lesions in pneumonia.High production of TNF-? may relate to the severity of pneumonia.There have already been several studies examining the association between pneumonia and single nucleotide polymorphisms(SNPs) that affect cytokine productivity.SNPs of TNF-?,-1 031,-863,-857,-308 and-238 have been identified.The variant alleles of these SNPs have suggested to be related to high TNF-? production and the severity of pneumonia.Therefore,the aim of this study is to examine the association between the severity of pneumonia in Chinese and the following SNPs: five in the TNF-? gene promoter(-1 031,-863,-857,-308,-238).METHODS: A total of 117 Chinese individuals were enrolled in this study.They were 67 patients with pneumonia and 50 healthy subjects.TNF-? was genotyped by polymerase chain reaction-restriction fragment length polymorphism for all subjects.The frequency distributions of genotypes in different groups were analyzed by SPSS 11.5 program.RESULTS: Frequency of subjects who carried at least one variant allele in TNF-?-1 031,-863,-857,-308,-238 SNPs among pneumonia patients was significantly higher than that in healthy subjects.And frequency of subjects who carried variant allele in TNF-?-863,and-308 SNPs among severe adult community acquired pneumonia patients was significantly higher than that in common pneumonia patients.CONCLUSION: TNF-?-863 and-308 SNPs appear to be associated with severe adult community acquired pneumonia in Chinese populations.

To intensify the medical students' clinical skills training is the main objective of medical education,and it can promote the reform of clinical teaching.In the whole process of clinical practice,we have adopted means of teaching before receiving clinical practical training,estimating indexes of examine,making use of diversified clinical skill training,supervising clinical practical quality.The operation ability of medical students has greatly increased.

Objective To develop a rapid, accurate, specific method to detect causative agent of hand, foot and mouth disease (HFMD). Methods Specific primers and probe were designed based on highly conserved VP1 region of enterovirus 71, coxsackie virus A16 and enterovirus. The sensitivity and specificity of the real-time RT-PCR was evaluated with 35 stool samples collected from pediatric patients with suspected HFMD and 20 clinical samples from health pediatric patients. Results Out of 35 clinical samples from suspected HFMD, 35 samples were identified as positive for enterovirus, 25 clinical samples were identified as positive for enterovirus 71, 8 clinical samples were identified as positive for coxsackie virus A16, among which 3 clinical samples were identified as positive for enterovirus 71 and coxsackie virus A16. The clinical diagnostic accordance rate is 85.71%. Out of 20 clinical samples from normal pediatric patients, 5 clinical samples were identified as positive for enterovirus, 20 clinical samples were negative for enterovirns 71 and coxsackie virus AI6. Conclusion Our results indicate real-time RT-PCR offers a rapid, sensitive, specific and cheap method to detect pathogen of HFMD from clinical specimens.

Objective To observe the effects of Bushen Huoxue Formula on free radical metabolism and p16 protein expression in heart of aged rats; To discuss its protective mechanism to the heart of aged rats.Methods One hundred were divided into young control group, the natural aging group,Bushen HuoxueFormula high-, medium- and low-dose groups, with 20 rats in each group. Each medication group was given relevant medicine for gavage, once a day for 16 weeks. 1 hour after the last administration, after the rats were sacrificed, serum and heart were taken. The contents of NO and MDA and activities of CAT and SOD in serum and myocardial,β-galactosidase enzyme and p16 protein expression in myocardial tissue were detected.ResultsCompared with the natural aging group, NO content and SOD activity in the serum of rats inBushen Huoxue Formula high-dose group increased significantly (P<0.05) and MDA content in allBushen Huoxue Formula groups decreased (P<0.01); NO content, CAT and SOD activity in the myocardial tissue of rats inBushen HuoxueFormula high-dose group increased significantly, and MDA content decreased significantly (P<0.05). CAT activity in allBushen HuoxueFormula groups increased (P<0.05,P<0.01).β-galactosidase enzyme and p16 protein expression in myocardial tissue in allBushen Huoxue Formula groups decreased.Conclusion Bushen Huoxue Formula can in hibit the aging of myocardial-aged rats, and the mechanism might be related to its anti-oxidative damage and inhibition of tumor suppressor gene p16 expression.

Objective:To assess the effect of accelerated aging on the color and opacity of veneers bonded resin.Methods:28 discs were made with IPS e.max Press A1 shade with the thickness of 0.4 mm.Ceramic discs were divided into 4 groups(n=7). BEAUTIFIL Flow A1 ,DEO-Link SE Transparent,Panavia F Light color and Viriolink N transparent resin cements were applied on the porcelain discs with a thickness of 0.1 mm.Curing was performed with a calibrated LED curing-light for 40 seconds.Color and opac-ity differences of the porcelain substructures after 65 h(150 kJ/m2 )of UV ageing test were examined with a colorimeter.The results were analysed statistically with One-way ANOVA,LSD(L)test.Results:All samples showed significant changes in color and opaci-ty.The ΔE of all materials ranged from 0.6 to 2.29.DEO-Link SE showed the highest color change,while BEAUTIFIL Flow the lowest.The opacity changes ranged from 0.46 to 2.55 and its variation was not significant among groups except for Panavia F.Con-clusion:The accelerated ageing may lead to color changes of ceramic veneer bonded by resins in a clinically acceptable range(ΔE<3).

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