Vitreoscilla Hemoglobin(VHb), with the function of increasing the growth of and product yield by a heterologous host, has been widely use in the area of fermentation, environment protection, transgenic animal and plant, recombinant protein expression, etc. Fusion protein of VHb with other enzyme or protein can enhance activity and stability of the enzyme or isolation efficiency of the protein. The reconstitution of VHb will be helpful to obtain ‘novel’ proteins which have better activity.

Objective To evaluate the value of creatine kinase MB(CK-MB) and cardiac troponin I(cTnI)to earlier diagnosis on myocardial injury in newborn infants with asphyxial.Methods Dynamic variation of serum CK-MB and cTnI levels were measured at birth 1,5 and 10 days,respectively,in 40 asphyxia newborn infants and 20 control neonates.Results Serum CK-MB and cTnI levels of asphyxia neonates were significantly higher than those in control group(P0.05).Conclusion The determination of CK-MB and cTnI levels can help the prediction of myocardial injury after asphyxia.

Objective To compare the differences of clinical characteristics between genotype B and C chronic hepatitis B(CHB)patients and to summarize clinical factors related to genotype C hepa- titis B virus(HBV)infection.Methods Seventy eight CHB patients who were diagnosed with genotype B or C infection by liver puncture biopsy and genotyping were enrolled.Their serum HBV DNA levels were detected.Severe hepatitis,liver cirrhosis,hepatocellular carcinoma and HBeAg positive rate were analyzed to determine the pathologic inflammation and fibrosis degree of liver tissue.Chi square test and Logistic multiple regression analysis were employed for the statistical analysis.Results The serum albumin and pre-protein were lower in genotype C CHB patients than that in genotype B.The alanine aminotrans- ferase,total bilirubin and prothrombin time were higher in genotype C CHB patients than that in genotype B.The rates of genotype C patients increased significantly with the grade of liver necroin- flammation progressing from GO to G4(1.8%,11.1%,20.4%,33.3%,33.3%) and the stage of liver fibrosis progressing from SO to S4(5.6%,5.6%,14.8%,33.3%,40.7%),but the rates of genotype B patients did not change significantly with the grade of liver necroinflammation(16.7%, 25.0%,25.0%,20.8%,12.5%)and stage of liver fibrosis progressing(16.7%,29.2%%,20.8%, 16.7%,16.7%).There was statistical significance in grades of liver necroinflammation(X~2= 11.49,P=0.022)and stages of liver fibrosis(X~2=13.56,P=0.006)between genotype B and gen- otype C patients.The rates of genotype C CHB patients were higher than,similar with and lower than the rates of genotype B patients of HBV DNA level above 1.0?10~6 copy/mL,between 5.0?10~2-1.0?10~6 copy/mL and under 5.0?10~2 copy/mL,respectively(51.8% vs 12.5%,35.2% vs 45.8% and 13.0% vs 41.7%).There was statistical significance of HBV loads between genotype B and genotype C patients(X~2=13.25,P=0.001).HBeAg positive rate in genotype C patients was significantly higher than that in genotype B patients(61.1% vs 25.0%,X~2=8.67,P=0.003).The rates of decompensated cirrhosis,compensated cirrhosis and no-cirrhosis in genotype C patients were higher than,similiar with and lower than the rates in genotype B patients,respectively(40.7% vs 4.2%,22.2% vs 20.8% and 37.0% vs 75.0%).There was statistical significance of the rate of cirrhosis between genotype B and genotype C patients (X~2=12.47,P=0.002).Conclusions The degree of liver necroinflammation and fibrosis,the HBeAg positive rate and the incidence of cirrhosis are all related with genotype C HBV infection.

To improve the growth enhancement activity of Vitreoscilla hemoglobin(VHb), Vitreoscilla hemoglobin gene(vgb) was mutated by error-prone PCR and then reconstituted by DNA shuffling. The shuffling library was constructed by inserting the shuffled genes into the downstream of vgb natural promoter and transforming them into E.coli DH5?. Mutated active VHb proteins were first screened in test tubes according to host cell pellets color and then in shake flasks according to host pellets wet weight .One active mutant protein, VHb′042506, was obtained after second screening. It could increased the host wet weight by 31.25% and 58.75% than that of the control which bearing natural VHb under microaerobic and extremely microaerobic conditions, respectively. Sequencing and alignment results showed that 11 nucleotides were mutated, thus resulted in 4 amino acids changes occurred in this mutant protein. CO difference spectrum test also indicated that it had higher specific absorption.

BACKGROUNDTo clone and express the ss1 recombinant gene containing S gene and preS1 (10-50 AA) gene in P. pastoris expression system.

METHODSThe fusion gene ss1 containing the S (1-222 AA) gene and preS1 (10-50 AA) gene was constructed with PCR method. The fusion ss1 gene was cloned into the expression vector of pPIC3.5k. The linear vector DNA was transformed into the host cell of GS115 with electroporation method. After screening with G418, the product was induced to express with methanol and its antigenicity was analyzed.

RESULTSThe molecular weight of expressed ss1 protein was about 30,000 dalton. The product was reactive to anti-HBs and anti-preS1 mAb.

CONCLUSIONThe fusion gene was efficiently expressed in P. pastoris expression system.The expressed products have the antigenicity of both S and preS1 protein.

Blotting, Western ; Enzyme-Linked Immunosorbent Assay ; Gene Expression ; Hepatitis B Surface Antigens ; genetics ; metabolism ; Pichia ; genetics ; Plasmids ; genetics ; Protein Precursors ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transformation, Genetic

Epidemiology is a discipline characterized by complicated theory and practice.How to make the practice course function better is a topic worthy of exploring in educational reform for clinical students.The article explored the‘Student-Dominated’ Model based on ‘Problem-Based Learning ’ and ‘Team Based Learning ’ in teaching process and compared the model with the traditional one ( Teacher-Dominated Model) .Suggestions were given to further improve effectiveness of epidemiology practice courses.

ORF3 and partial ORF1 regions were amplified with RT-PCR f rom two patients (T1 and T11)infected with new genotype of hepatitis E Virus. Th e PCR products were cloned and sequenced. The results showed that G-C rich regi on in ORF3 was deleted when amplified with normal PCR reaction. However, PCR rea ction containing G-C melt solution can overcome this problem. The sequence anal ysis showed that T1 and T11 belong to a new genotype of HEV which differs from g enotype I,II and III reported.T1 and T11 have 79%～82%, 80%～81% and 83%～85% id entical to genotype I,II and III respectively.

AIMTo study preventive and therapeutic effect of zinc sulfate on lung injury during superior mesenteric artery occlusion (SMAO) shock and their mechanism of action.

METHODSModel of rabbit SMAO shock was made. The effect of zinc sulfate on the malondialdehyde (MDA) in erythrocyte membrane and plasma, oxidase (XOD) in plasma, superoxide dismutase (SOD) in erythrocyte and MDA, SOD and pulmonary surfactant (PS) in lung tissues homogenate were observed.

RESULTSThe administration of zinc sulfate decreased MDA and XOD, prevented the reduction of SOD and PS, and alleviated lung injury.

CONCLUSIONIt is suggested that lung is injured during SMAO shock and zinc sulfate possesses preventive and therapeutic effect, through stabilized membrane.

Animals ; Female ; Lung ; metabolism ; Lung Injury ; drug therapy ; etiology ; metabolism ; Male ; Mesenteric Artery, Superior ; pathology ; Mesenteric Vascular Occlusion ; complications ; drug therapy ; metabolism ; Rabbits ; Shock ; complications ; drug therapy ; metabolism ; Zinc Sulfate ; therapeutic use

OBJECTIVETo search the forming cause and the correlation between the clinical phenotype and chromosome band by the cytogenetic analysis on a case of ring chromosome 21 syndrome.

METHODSIdentification and location of 21 ring chromosome were performed with the G-banding, C-banding, N-banding, high-resolution banding and fluorescence in situ hybridization (FISH) techniques.

RESULTSIt was found that the karyotypes of the patient's parents are normal. The patient's karyotype is 46,XY, r(21)[91]/46,XY,r(21;21)(p11q22.3;p11q22.3) [5]/45,XY,-21[4].

CONCLUSIONThe clinical phenotype of ring chromosome 21 syndrome is related to the deletion of distal segment of 21q, and the abnormal sexual development of male is related with the deletion of 21q22.3.

Child, Preschool ; Chromosome Aberrations ; Chromosome Disorders ; genetics ; pathology ; Chromosomes, Human, Pair 21 ; genetics ; Cytogenetic Analysis ; methods ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Male ; Phenotype ; Ring Chromosomes ; Syndrome

Acetylcysteine ; pharmacology ; Animals ; Cell Line ; Gene Expression ; Lipopolysaccharides ; pharmacology ; Macrophages ; drug effects ; Mice ; Toll-Like Receptor 2 ; genetics ; Toll-Like Receptor 4 ; genetics