Child ; Gastroesophageal Reflux ; diagnosis ; therapy ; Humans

Bilirubin ; Child ; Duodenogastric Reflux ; diagnosis ; Esophageal Diseases ; diagnosis ; Esophagus ; metabolism ; Gastroesophageal Reflux ; diagnosis ; Humans

Adolescent ; Bilirubin ; metabolism ; Child ; Child, Preschool ; Duodenogastric Reflux ; metabolism ; Esophageal pH Monitoring ; Female ; Humans ; Male ; Stomach ; physiopathology

Objective To determine the influence of protein fusion on the biological characteris- tics of hymidine kinase(TK)and human immunodeficiency virus(HIV)Tat recombinant protein. Methods By utilizing polymerase chain reaction(PCR)technique,different fragments containing two,four or six glycines(Gly)were inserted between the HIV Tat gene and TK,and cloned into PBK vector.After testified by sequencing,the vectors were transfected into E coli.After induced by iso- propyl thiogalactose(IPTG),bacilli were collected and destructed by ultrasonic,the fusion proteins were determined by monoclonal antibody against HIV protein.HepG2 cells were incubated in DMEM supplement with 10?g/mL HIV-Gly(n)-TK(n=0,2,4,6)fusion protein,TK-HIV Tat and only HIV Tat.HepG2 cells in different groups were detected by immunofluorescence assay 24 hours after transduction with HIV Tat monoclonal antibody.The rate of apoptosis after cells were incubated with gencilovir(10?g/mL)for 3 days was determined by cell flow cytometry,while survival cell ratio was recorded by trypan blue.The data were analyzed by statistics(t-test).Results The Tat-Gly(n)-TK (n= 0,2,4,6)recombinant genes were constructed and inserted into PBK vectors,which were expressed in E coli and then purified.Cells in different groups,which were incubated with Tat-Gly (n)-TK(n=0,2,4,6)fusion proteins,Tat-TK fusion protein,TK-Tat fusion proteins or only Tat proteins respectively,were detected by immunofluorescence assay.The intensities of fluorescence in different groups were almost same,but the ratios of cell survival or apoptosis were different.The highest ratio of cells apoptosis(14.77%)was in the group that cellular culture medium was mixed with Tat-Gly(4)-TK fusion protein,followed by the groups containing 6,2 glycines or no TK gene in genes(4.30%,12.69% and 1.03%,respectively).There were significant differences between each 2 groups among the all groups(t-test,P

Dipeptides ; administration & dosage ; therapeutic use ; End Stage Liver Disease ; drug therapy ; virology ; Female ; Hepatitis B virus ; Humans ; Male

The recombinant fusion protein staphylokinase-hirudin(rSFH) was purified from the high density-fermented engineered E.coli by means of ion-exchange chromatography (IEC) and gel filtration (GF). The purity of rSFH reached to more than 98% determined by RP-HPLC and SDS-PAGE, and the yield was up to 0.7g per liter of fermentation broth. The analysis of homologous dimmer of rSFH appeared during the purification and calculation of the surface hydrophobic area had been carried out by means of hydrophobic chromatography and MALD-TOF. The influence of sodium chloride and temperature on the behavior of rSFH reversible dimerization was analyzed by high performance sized- exclusive chromatography(HPSEC). It is concluded that the hydrophobic interaction played an important role in the reversible dimerization of rSFH.

OBJECTIVETo investigate the association between the chronic hepatitis B cirrhosis and HLA-DRB1 * 1301,1302 gene.

METHODSHLA-DRB1 * 1301,1302 allele in 27 patients with chronic hepatitis B cirrhosis and 30 patients with chronic hepatitis B was analyzed by using the polymerase chain reaction/sequence specific primer (PCR-SSP) technique.

RESULTSThe frenquency of HLA-DRB1 * 1301,1302 allele in the chronic hepatitis B cirrhosis group was markly higher than that in the chronic hepatitis B group.

CONCLUSIONHLA-DRB1 * 1301,1302 is closely associated with the suseptibility to chronic hepatitis cirrhosis.

Adult ; Alleles ; Female ; HLA-DR Antigens ; genetics ; HLA-DRB1 Chains ; Hepatitis B, Chronic ; genetics ; Humans ; Liver Cirrhosis ; genetics ; Male

0.05);however there were significant difference between D4T+DDI+NVP group and AZT+DDI+NVP group(P

OBJECTIVETo establish a mathematical model of hepatitis C virus (HCV) replication and develop a working theory for antiviral therapy in order to understand the dynamics of HCV replication.

METHODSPeripheral blood cells of 4 hepatitis C patients were cultured. Quantities of the HCV were detected every 15 min by real-time PCR. The data were analyzed using SPSS software. A mathematical functional relationship between HCV RNA and the time lapse was established.

RESULTSThe quantity of HCV RNA declined and it fell into a mathematical model: Y=3E+0.8e(-0.5467x) (r=0.9547). The estimated virion half-life was 45 min on the average.

CONCLUSIONSThe decline of HCV RNA in the blood is not of a linear trend and the HCV RNA lasts a longer time although the speed of the decline is faster than that in vivo.

Adult ; Half-Life ; Hepacivirus ; Hepatitis C, Chronic ; blood ; virology ; Humans ; Models, Theoretical ; Nonlinear Dynamics ; RNA, Viral ; blood ; Viral Load ; Virus Replication

OBJECTIVETo study the association between HLA-DRB1 gene polymorphism and severe chronic hepatitis B.

METHODS26 patients with severe chronic hepatitis B were investigated for HLA-DRB1 gene polymorphism by polymerase chain reaction-sequence specific primers technique. The results were compared with those from 45 normal healthy people by use of chi2-test of Microsoft SPSS 13.0.

RESULTSThe frequency of HLA-DRB1 * 1301/1302 allele in severe chronic hepatitis B group was significantly higher than the frequency in the control group, while the frequencies of HLA-DRB1 * 1201/1202, 1501/1502 allele were not significantly different.

CONCLUSIONHLA-DRB1 * 1301,1302 is closely associated with the suseptibility to severe chronic hepatitis B, While HLA-DRB1 * 1201/1202, 1501/1502 have no association with severe chronic hepatitis B.

Adult ; Case-Control Studies ; Female ; HLA-DR Antigens ; genetics ; HLA-DRB1 Chains ; Hepatitis B virus ; physiology ; Hepatitis B, Chronic ; genetics ; pathology ; virology ; Humans ; Male ; Middle Aged ; Polymorphism, Genetic ; Severity of Illness Index ; Young Adult