1.DNA-mediated immunization to hepatitis B surface antigen: effects of a plasmid coexpressing hepatitis B surface antigen and interleukin 18
Hengning KE ; Chongwen SI ; Jun CHENG
Chinese Journal of Infectious Diseases 2000;0(02):-
Objective To observe the specific humoral and cellular immune response in BALB/c mice injected with pS and p18S. Methods pS and p18S were constructed separately by inserting HBsAg gene fragment and the fusion gene fragment of HBsAg and mouse interleukin 18(IL 18) into the reading frame of pcDNA3.1+. Mice were injected with either plasmid intramuscularly in a total dose of 300 ?g per mouse. Every serum sample was detected for anti HBs using enzyme linked immunosorbent assay(ELISA). Furthermore, HBsAg specific cytotoxic T lymphocytes activity was measured. Results The expression of HBsAg was demonstrated by ELISA in p815 cells transfected with pS and p18S. pS can stimulate a positive antibody response. The average level was 135 mIU/ml, with the highest level of 530 mIU/ml. p18S could elicit relatively lower antibody response which was 20 mIU/ml. HBsAg specific CTL activities were 37.1% and 34% separately in pS and p18S immunized mice. It was only 13.2% when detected in pcDNA3.1+ immunization. Conclusion pS is effective to stimulate a humoral and cellular response in H 2d mice. IL 18 gene can not enhance the immune response when fused with HBsAg gene. Conversely, it seems to inhibit an immune response.
2.The detection of tumor necrosis factor-? and interleukin-6 in the ascitic fluid of cirrhotic patients compli cated.with spontaneous bacterial peritonitisand its significance
Xiaohong LIN ; Hongli XI ; Chongwen SI
Chinese Journal of Practical Internal Medicine 2001;0(03):-
Objective To investigate the clinical significance of tumor necrosis factor-?(TNF-?)and interleukin-6(IL-6)in the ascitic fluid of cirrhotic patients with spontaneous bacterial peritonitis(SBP).Methods TNF-? and IL-6 in the ascitic fluid of 32 cirrhotic patients with SBP were measured by ELISA.The group was compared with the value in transudatory ascites of 30 cirrhotic patients.Results The levels of TNF-? and IL-6 in the ascitic fluid of cirrhotic patients with SBP were much higher than those in transudatory ascites of cirrhotic patients(P
3.Predictors of response in patients with HBeAg-positive chronic hepatitis B with adefovir dipivoxil treatment
Jun LI ; Hong ZHAO ; Chongwen SI
Chinese Journal of Practical Internal Medicine 2001;0(06):-
Objective To investigate the effective predictors of response in HBeAg-positive patients treated with adefovir dipivoxil(ADV)and to provide evidence for individualized treatment.Methods Patients administered with ADV for 48 weeks in a randomized,placebo-controlled,multicenter trial were studied.Statistical analyses,such as Backward stepwise logistic regression and 2?2 method were used for predictors analysis at week 48.Results The baseline serum ALT levels,HBV DNA levels,and undetectable serum HBV DNA by PCR at week 24 were predictors for HBV DNA negativity at week 48.The median of serum ALT levels and HBV DNA levels prior to treatment were 134.5 U/L and 6.57 lg copies/mL,respectively.Patients with baseline ALT levels higher than the median,HBV DNA levels lower than the median,and serum HBV DNA undectectable by PCR at week 24 had greater rate of HBV DNA negativity(93.3%),HBeAg loss(60%)and HBeAg seroconversion(40%)at week 48 than the others.47.8% of patients whose HBV DNA levels were positive at week 24 also achieved HBV DNA negativity at week 48,and 8.6% achieved HBeAg seroconversion.Conclusion Better response at week 48 has significantly higher serum ALT levels and lower HBV DNA levels prior to treatment and HBV DNA negativity at week 24 compared with non-response.Patients whose HBV DNA levels ware still positive at week 24 should continue therapy.
4.Therapeutic efficacy of 18 α-Diammonium glycyrrhizinate phosphatidylcholine complex in patients with chronic virus hepatitis
Hong ZHAO ; Yong CHEN ; Jiazhang XU ; Jun WANG ; Chongwen SI
Clinical Medicine of China 2008;24(8):770-772
Objective To investigate the therapeutic effieacy and safety of 18 α-Diammonium glycyrrhizinate phosphatidylcholine complex (DGPC) in patients with chronic hepatitis B and or C with elevated aminotransferase. Methods 55 patients with chronic hepatitis B and or C, with serum alanine aminotransferase (ALT) of 2 to 10 times the upper limit of normal were randomly assigned to receive DGPC or Diammonium glyeyrrhizinate (DG) for 12 weeks. Then they were followed up for an additional 4 weeks. From week 1 to 10, DGPC or DG was given as 150 nag,three times a day (TID). At the 11th week,the drug was given as 100 mg,TID. Then 50 mg,TID for the 12th week. Results ALT was markedly decreased after receiving DGPC 4,8,12 weeks (P=0.00). ALT normalization rate at the end of therapy was similar (38.5% vs 34.5% ,P =0.76). Drug-related adverse events were similar. Conclusion DGPC can rapidly and safely decrease aminotransferase in patients with chronic viurs hepatitis.
5.Evaluation of specificity and sensitivity of three kinds of real-time PCR instruments
Zheng ZENG ; Dan LIU ; Weibo GONG ; Chongwen SI
Chinese Medical Equipment Journal 2003;0(10):-
Objective:To evaluate the specificity and sensitivity of ABI 7000,7300 and 7500 real-time PCR instruments.Methods:Random,double-blind clinical trial;We selected 30 patients with HBsAg and HBV DNA positive for positive group and 30 health volunteers with HBsAg and HBV DNA negative for negative group.We also selected three of HBV DNA positive and negative samples for repeat test.Results:The 30 samples of positive group were detected HBV DNA.The 30 samples of negative group were not detected HBV DNA.However,in positive group,the HBV DNA levels were different.The related index of ABI 7000 with ABI 7300,ABI 7500 and ABI 7300 with ABI 7500 was 0.89,0.89 and 0.98,respectively.The HBV DNA level was highest detected with ABI 7300 instrument,and then with ABI 7000 instrument.The lowest level was detected with ABI 7500 instrument.There was significance among these instruments(p
6.Clinical effect of consensus interferon in the retreatment of previous nonresponders or relapsers of chronic hepatitis C
Xiaojuan LI ; Yanyan YU ; Chongwen SI ; Al ET ;
Chinese Journal of Infectious Diseases 2001;0(05):-
Objective To evaluate the efficacy and safety of consensus interferon in previous nonresponders or relapsers of chronic hepatitis C as well as to compare the correlation between viral kinetics and the efficacy. Methods 32 patients who are previous nonresponders or relapsers to IFN received CIFN 15?g three times a week for 24 weeks. Efficacy was assessed by ALT and HCV RNA (viral kinetics) during the 24 weeks of treatment period and 6 month follow up post treatment. Results After 24 weeks treatment, HCV RNA negative rate in the previous nonresponders and the previous relapsers were 90% versus 45.5% ( P 0.05 ). The major adverse events were fever and leukopenia. Conclusions CIFN 15?g as a monotherapy significantly improved end of treatment response rate in chronic hepatitis C patients. Baseline serum HCV RNA load and early virus kinetics are predictive parameters of SR.
7.Cloning and analysis of the genomic DNA sequence of augmenter of liver regeneration from rat.
Jing DONG ; Jun CHENG ; Qinhuan WANG ; Shuangshuang SHI ; Gang WANG ; Chongwen SI
Chinese Medical Sciences Journal 2002;17(2):63-67
OBJECTIVETo search for genomic DNA sequence of the augmenter of liver regeneration (ALR) of rat.
METHODSPolymerase chain reaction (PCR) with specific primers was used to amplify the sequence from the rat genome.
RESULTSA piece of genomic DNA sequence and a piece of pseudogene of rat ALR were identified. The lengths of the gene and pseudogene are 1508 bp and 442 bp, respectively. The ALR gene of rat includes 3 exons and 2 introns. The 442 bp DNA sequence may represent a pseudogene or a ALR-related peptide. Predicted amino acid sequence analysis showed that there were 14 different amino acid residues between the gene and pseudogene. ALR-related peptide is 84 amino acid residues in length and relates closely to ALR protein.
CONCLUSIONThere might be a multigene family of ALR in rat.
Amino Acid Sequence ; Animals ; Base Sequence ; Cloning, Molecular ; DNA ; genetics ; Female ; Humans ; Liver Regeneration ; genetics ; Male ; Mice ; Molecular Sequence Data ; Polymerase Chain Reaction ; Proteins ; genetics ; Pseudogenes ; genetics ; Rats
8.Preliminary investigation on the relation between clinical progress and anti-small monomolecular peptides antibody in individual infected with HIV.
Xiaoyuan XU ; Huichun XING ; Weibo GONG ; Hongmei CHEN ; Chongwen SI ; Yan WANG ; J C CHERMANN
Chinese Journal of Experimental and Clinical Virology 2002;16(3):286-287
OBJECTIVETo study the quantity of anti-R7V in individuals infected with HIV and AIDS patients and its relation with the progression of disease.
METHODSELISA and precipitation and other methods were used to investigate the quantity of anti-R7V in asymptomatic long-term survivors and AIDS patients.
RESULTSPositive rate and quantity of anti-R7V were higher in the HIV active ones and AIDS. It showed that the quantity and positive rate of anti-R7V were rather high in dissolving test.
CONCLUSIONSIt is strong suggestion for anti-R7V to obstruct the replication of virus by interfering the connection between HIV with CCR5 or CXCR4 and so it impossible HIV entering to CD4+ T cells.
Acquired Immunodeficiency Syndrome ; immunology ; Adolescent ; Adult ; Aged ; Child ; Female ; Follow-Up Studies ; HIV Antibodies ; blood ; HIV Infections ; immunology ; HIV Long-Term Survivors ; HIV-1 ; immunology ; Humans ; Male ; Middle Aged ; Receptors, CCR5 ; physiology ; Receptors, CXCR4 ; physiology
9.Lamivudine-interferon alpha 2b sequential therapy versus lamivudine monotherapy in HBeAg-positive chronic hepatitis B patients
Jinghang XU ; Yanyan YU ; Chongwen SI ; Xinyue CHEN ; Zhonghou HAN ; Yong CHEN ; Wenjin ZHANG ; Daozhen XU ; Yuping CHEN ; Xueying LI ; Jie QIU
Chinese Journal of Infectious Diseases 2012;30(6):354-358
Objective To compare the efficacy and safety of Iamivudine-interferon sequential therapy and lamivudine monotherapy in HBeAg-positive chronic hepatitis B (CHB) patients.MethodsA total of 172 patients with HBeAg-positive CHB were randomized to sequential group (n=83) or lamivudine group (n=89).Sequential group were administrated with lamivudine 100 mg/d and 5 million units interferon alpha 2b subcutaneous injection every other day for 24 weeks were added since week 25 of treatment.Lamivudine group were administrated with lamivudine 100 mg/d for 48 weeks.All subjects were followed up for 24 weeks after drug withdrawal.Measurement data with homogeneity of variance were analyzed by using t test and data with heterogeneity of variance were analyzed by using rank sum test.The comparison of rates was done by chi square test or Fisher exact test.ResultsThe baseline hepatitis B virus (HBV) DNA levels of patients in sequential group and lamivudine group were (7.8±1.0) and (7.9±1.1) lg copy/mL,respectively (P>0.05),and the baseline alanine aminotransferase (ALT) levels were (210.5 ± 150.1 ) and (211.9 ± 160.9) U/L,respectively (P>0.05).At the end of treatment,higher ALT levels [(78.4±146.1) vs (36.1±32.4) U/L,P<0.05)] and HBV DNA levels [(4.5±1.5) vs (3.8±1.3) lg copy/mL,P<0.05)] levels,lower response rates (65.8% vs 83.5%,P<0.05),and similar HBeAg loss rates (31.6% vs 22.2%,P>0.05) and HBeAg seroconversion rates (27.6% vs 16.0%,P>0.05) were found in sequential group compared with lamivudine group.At the end of follow-up,higher ALT levels [(126.0±143.1) vs (82.7±83.0) U/L,P<0.05)],similar HBV DNA levels [(5.3±1.5) vs (5.0±1.5) lg copy/mL,P>0.05)],similar HBeAg loss rates (25.0% vs 32.3%,P>0.05) and HBeAg seroconversion rates (25.0 % vs 26.2 %,P>0.05) were found in sequential group compared with lamivudine group.YMDD motif mutation rate in sequential group was lower than lamivudine group at week 48 of treatment (10.5% vs 26.9%,P<0.05).ConclusionsLamivudine-interferon sequential therapy and lamivudine monotherapy are both effective in HBeAg-positive CHB patients,while HBV mutations are reduced in patients with sequential therapy.
10.Quasispecies and variations of hepatitis B virus: core promotor region as an example.
Jing DONG ; Jun CHENG ; Qinhuan WANG ; Gang WANG ; Shuangshuang SHI ; Yan LIU ; Xiaobing XIA ; Li LI ; Guoqing ZHANG ; Chongwen SI
Chinese Journal of Experimental and Clinical Virology 2002;16(3):264-266
OBJECTIVETo investigate the HBV quasispecies groups in the patients with chronic HBV infection.
METHODSA set of specific primers was synthesized according to DNA sequence of HBV strain found in China. The whole core promoter (CP) region was amplified by PCR method from the sera of 3 patients with chronic HBV infection, and the PCR products were subcloned into pGEM Teasy vectors. The clones were randomly selected to be sequenced. Sequence comparison of the selected clones was made to find the difference.
RESULTSBy comparison, it was found that each sequence of selected clones was different. The point mutation always occurred in TATA-like boxes, especially from T to C replacement on 184 site. There is a hot region (33.3% 5/15) in basic core promoter where deletion mutation frequently happened.
CONCLUSIONSThere is a hot deletion region near DR I in CP. The replacement at 184 nt (T to C) in the third TATA-like box may influence the expression of pre-C/C protein. The sequencing results suggest that there are HBV quasispecies groups in chronically infected patients.
DNA, Viral ; analysis ; Genes, Viral ; genetics ; Genetic Variation ; Hepatitis B Core Antigens ; genetics ; Hepatitis B virus ; classification ; genetics ; Hepatitis B, Chronic ; virology ; Humans ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; TATA Box ; genetics