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 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 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 To compare the virological responses between Peg IFNαt-2a and Peg IFNα-2b in treatment of chronic hepatitis C.Methods Clinical data of 46 chronic hepatitis C (CHC) patients were retrospectively reviewed.Patients were divided into two groups:Peg IFNα-2a group ( n =24) was given Peg IFNoα-2a 180 μg/week and ribavirin; Peg IFNα-2b group (n =22 ) was given Peg IFNα-2b1.5 μg · kg-1 · week-1 and ribavirin.Serum HCV RNA load at 4th,12th and 24th week of the treatment were detected to evaluate the virological responses.Softwares SPSS 13.0 and PEMS 3.1 were used for statistical analysis.Results There was no significant difference in rapid response rate,early response rate,complete response rate and non-response rate between the two groups (x2 =0.689,0.105,0.105 and 0.105,P ＞ 0.05 ).However for patients with high viral load ( HCV RNA ＞ 6 logl0 copies/mL) at baseline,the rapid response rate in Peg IFNα-2a group ( 86.67％ ) was higher than that in Peg IFNα-2bgroup (42.86％ ) (x2 =4.365,P ＜ 0.05 ).Conclusion Peg IFNα-2a combined with ribavirin may have higher rapid response rate than that of Peg IFNα-2b combined with ribavirin in CHC patients with high viral

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 Obstructive sleep apnea hypopnea syndrome (OSAHS) often causes damage to multiple systems, especially to the central nervous system, inducing cognitive dysfunction.This study aims to explore the possible correlation of the expressions of serum hypoxia-inducible factor-1α (HIF-1α) and brain-derived neurotrophic factor (BDNF) with cognitive impairment in rats under different hypoxia conditions.Methods Twenty-four Wistar rats were equally randomized into a normal control, a chronic intermittent hypoxia (CIH), and a chronic continuous hypoxia (CCH) group.The rats of the CIH group were placed in a hypoxia chamber filled with N2 and air, the oxygen concentration switched from (7±0.5)% to 21%, 1.5 minutes for each state and 4 minutes for each cycle, while those of the CCH group were placed in another hypoxia chamber with the oxygen concentration of (7±0.5)%, 8 hours a day and all for 30 days.Then we recorded the body weight of the rats, detected the expressions of serum HIF-1α and BDNF by ELISA, and observed the changes of behavior by Morris water maze test and those of the hippocampal morphological structure by HE staining.Results At 30 days after modeling, the body weight of the rats was significantly decreased in the CIH and CSH groups as compared with the normal control ([195.75±6.497] and [180.88±12.017] vs [218.63±15.287] g, P<0.05).Positioning navigation showed that the escape latency was significantly longer in the hypoxia models than in the controls (P<0.05), even longer in the CIH than in the CCH group (P<0.05).Spatial exploration test manifested a lower frequency of crossing the platform in the CIH and CCH groups than in the control ([2.63±1.45] and [3.22±1.30] vs [4.97±0.47] times, P<0.05).The expression levels of serum HIF-1α and BDNF were significantly higher in the CIH ([36.14±9.34] and [1625.34±332.44] pg/mL) and CCH ([27.27±6.88] and [1204.07±363.81] pg/mL) than in the normal control group ([14.11±4.06] and [1036.40±124.48] pg/mL) (P<0.05), even higher in the CIH than in the CCH group (P<0.05).HE staining exhibited scattered and disorderly arrangement of hippocampal neurons in the model rats, with unclear nuclear membrane, pyknosis of the nuclei, darkly stained cytoplasm, and some damaged cells.More obvious absence and vacuolization of some cells were observed in the rats of the CIH group.Conclusion Chronic hypoxia inhibits the growth and development of rats and induces cognitive dysfunction.High-level HIF-1α in chronic intermittent hypoxia indicates hypoxia-stress of the body, while compensatory increase of serum BDNF may be involved in neuronal cell damage regulation.

Objective To investigate the change of the content of polysaccharides in crude and processed Fructus Corni,thus to approach the process mechanism of Fructus Corni.Methods The content of polysaccharides in the crude and processed Fructus Corni was determined by phenol-sulfuric acid method.Results After processing with wine,the content of polysaccharides of Fructus Corni decreased from 10.12 %(content of total polysaccharidesbeing 51.41 %) to 5.91 %(content of total polysaccharidesbeing 53.10 %),decreased by 41.60 %as compared with that in the crude Fructus Corni.Conclusion After processing with wine,the content of polysaccharides decreases markedly.The results provide certain evidence for approaching the process mechanism of Fructus Corni.