[Show abstract][Hide abstract] ABSTRACT: A nation-wide hepatitis B vaccination program was implemented in China starting in 1992. To study the change of hepatitis B virus (HBV) variant prevalence with massive immunization, large surface protein (LHBs) genes from the HBV surface antigen (HBsAg) positive sera were amplified and sequenced. The prevalence of LHBs mutants were compared between the 1992 and 2005 surveys in children and adult groups. The prevalence of 'α' determinant mutants in the children increased from 6.5% in 1992 to 14.8% in 2005, where G145R mutant occurred most frequently. In contrast, mutation frequencies showed little difference between 1992 (9.4%) and 2005 (9.9%) in adults. Moreover, compare to 1992 survey, the children groups surface (S) protein mutation frequency specifically increased (p=0.005) in 2005 survey, but not the Pre-S region (p>0.05). However, the adult groups mutation frequency increased in both pre-S and S region. Furthermore, the adult groups' disease related mutations pre-S2 deletion and start codon mutation were significantly higher than children groups in both the 1992 and 2005 survey (p<0.01). The massive immunization enhances the whole HBV S protein mutation; the prevalence of LHBs mutants, particularly disease related mutants, tends to increase with patient age.
[Show abstract][Hide abstract] ABSTRACT: To prepare monoclonal antibodies (McAbs) against human programmed death-1 ligant 1(PD-L1) and identify its bioactivity.
We immunized the BALB/c mice with Thioredoxin-(PD-L1) recombination protein which expressed by prokaryotic system. Prepare hybridoma cell by hybridoma technology and used enzyme-linked immunosorbent assay(ELISA) and Western-blotting assays to select positive hybridoma identify cell. Competition inhibition ELISA was carried out to identify the special bioactivity of antibody.
4 hybridoma cell strains which could secrete anti-(PD-L1) antibodies stably were selected. The McAbs has good affinity with its receptor. Purify anti-(PD-L1) with title 1:32 000 was obtained after large quantity preparation. At the same time we obtained 1 cell stain which could secret special anti-Trx McAbs.
We obtained anti-(PD-L1) McAbs with good bioactivity successfully, which lay the foundation for further study.
Zhonghua shi yan he lin chuang bing du xue za zhi = Zhonghua shiyan he linchuang bingduxue zazhi = Chinese journal of experimental and clinical virology 10/2010; 24(5):380-2.
[Show abstract][Hide abstract] ABSTRACT: In China, hepatitis is a huge public health problem. Outbreaks of hepatitis A are the most frequent cause of acute hepatitis, and to date, few epidemiologic investigations or molecular surveillance studies have been performed.
In 2006, two major outbreaks of hepatitis A occurred, one in Guigang City, southern China, and the other in Hetian City, northwestern China. Field and molecular epidemiologic investigations were conducted.
In Guigang, a single outbreak occurred in a school; 35 patients and 25 asymptomatic individuals were infected with 1 strain of hepatitis A virus (HAV). A case-control study showed that contaminated water was the likely transmission source. In Hetian, the epidemic of hepatitis A consisted of sporadic, small outbreaks involving as many as 20 wild HAV strains. A molecular epidemiology approach allowed us to identify two groups infected by individual HAV strains. Further fieldwork and a case-control study showed that ice cream was the suspected transmission source in one group. Our molecular epidemiology study showed that genetic variability between the HAV strains isolated from Guigang and Hetian and previously reported HAV strains was at least 4.3%.
Contaminated water and suspected ice cream were associated with outbreaks of hepatitis A. Viral genetic analysis may advance field investigations in complex situations.
Hepatology International 07/2009; 3(2):356-63. · 2.64 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: To construct the programmed cell death 1 ligant 1 (PD-L1) recombination expression vector, express the fusion protein in prokaryotic and analyze the biological action of express product.
The whole PD-L1 gene sequence was synthesized after codon optimized. Construct the thioredoxin-(PD-L1) recombination expression vector and express the fusion protein in E. coli. Purified the target protein and analyze the conjugated ability of protein by ELISA.
The PD-L1 recombinant expression vector has been constructed correctly. The target protein has been obtained with which expressed in high efficiency and production. The target protein can conjugate specifically with the PD-1, its specific receptor.
We have obtained the PD-L1 recombinant protein success with high biological activity. The result provide the basic condition for further study on antibody and mutually action between PD-L1 and chronic virus infectious.
Zhonghua shi yan he lin chuang bing du xue za zhi = Zhonghua shiyan he linchuang bingduxue zazhi = Chinese journal of experimental and clinical virology 02/2009; 23(1):5-7.
[Show abstract][Hide abstract] ABSTRACT: To determine the "alpha"dominant mutation of hepatitis B virus (HBV) in community-based Zhengding. Analysis the role of the newborn hepatitis B vaccination on the mutation.
Based on the national surveillance of hepatitis B, 11,478 people's sera were collected and tested by SPRIA with kits. Collect people's sera with positive HBsAg and amplify the S gene. Sequencing and clastwaling them with the standard sequences.
Overall, HBV DNA was successfully amplified and sequenced in 434 of 443 samples. 6.7% samples mutated in HBV "alpha" dominant region. The difference between the mutation ratio of the two loops of HBV "alpha" dominant between the people born before and after the year 1986 has no significance.
There were HBV "alpha" dominant mutant virus in the local area with a low infection rate in the population born after the year 1986. It could not explain the newborn hepatitis B vaccination can induce the prevalence of the "alpha" dominant mutate HBV.
Zhonghua shi yan he lin chuang bing du xue za zhi = Zhonghua shiyan he linchuang bingduxue zazhi = Chinese journal of experimental and clinical virology 02/2009; 23(1):11-3.
[Show abstract][Hide abstract] ABSTRACT: To explore the distribution of HBV genotype and serotype from Tibetan in Tongde, Qinghai.
Nested polymerase chain reaction (nPCR) was used for amplification of S gene and C gene of HBV from sera carried by Tibetan chronic HBV carrier in Tongde, Qinghai, then the HBV DNA positive products were sequenced by direct sequencing. Genotype and serotype were identified by analysis of sequence result.
271, which come from 311 sera samples with positive HBsAg randomly selected from natural community, were amplified and sequenced in both S gene and C gene successfully, 10 (3.7%), 261 (96.3%) out of them were identified as genotype C, recombinant between genotypes C and D respectively; 259 (95.6%), 10 (3.7%), 2 (0.7%) belonged to serotype ayw2, adr, adw2 respectively.
The recombinant between genotypes C and D was the main genotype in Tibetan chronic carrier with hepatitis Bin Tongde, Qinghai; the serotype of this areas was consisted largely of ayw2.
Zhonghua shi yan he lin chuang bing du xue za zhi = Zhonghua shiyan he linchuang bingduxue zazhi = Chinese journal of experimental and clinical virology 02/2009; 23(1):2-4.
[Show abstract][Hide abstract] ABSTRACT: Although significant advances have been made on the studies of HCV glycoproteins (E1 and E2) recently, the role of the peptides preceding each glycoprotein remains unclear. We expressed E1 and E2 using two individual plasmids to form HCV pseudoparticles (HCVpp) in order to characterize the peptides preceding E1 and E2. Our data show that 14 amino acids from the HCV core and 12 amino acids from the E1 C-terminus are required for E1 and E2 function, respectively. The lack of a long enough peptide preceding E1 or E2 will abolish HCVpp infectivity, and the presence of fewer than 14 amino acids ahead of E1 and 12 amino acids ahead of E2 may alter their glycosylation. Furthermore, the peptides preceding E1 and E2 may be interchanged or may be replaced by those from genotype 2a. Our findings may contribute to the future development of new anti-HCV drugs.
Biochemical and Biophysical Research Communications 12/2008; 378(1):118-22. · 2.28 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: An aberrant genotype of hepatitis B virus was discovered from a female child when we surveyed the status of the virus' infection in Guangxi of China. The full-length genome was amplified and sequenced. The length of genome is 3215 bp and the serotype of the virus is adr. In phylogenetic tree analysis with the standard genotype sequence of GenBank, the genome was clustered with genotype C, however, phylogenetic tree analysis of the individual segment supported recombination strain was formed. The segment between nt 1630 and 2880 was similar to genotype C, and the other part of genome close to genotype A. The result suggests it is a recombinant virus strain. The finding provides a reference to study the genotype and evolution of hepatitis B virus in China.
Bing du xue bao = Chinese journal of virology / [bian ji, Bing du xue bao bian ji wei yuan hui] 08/2008; 24(4):255-9.
[Show abstract][Hide abstract] ABSTRACT: In order to investigate the characterization of mutation and genotype distributing in the younger group which was under the universal vaccination. The sequence of HBV was analyzed to offer the information to control and prevention in the area.
Young person's sera with positive HBsAg are collected, and the Large S sequence of HBV including preS and S gene are amplified and sequenced. The genotype and serotype were determined by clastwal with the standard genotype sequence. And one virus complete genome is amplified.
The virus gene are successful amplified from the 33 sera. The sequence result indicate the 30 of 33 (90.9%) HBV genotype is B and 3 of 33 (9.0%) is C. The HBV serotype including ayw (1), adr (3), adw (29), 5 of 33 mutated in the "a" dominant of HBV, and the percentage is 15.2% . The HBV full length gene of serum number of 5856 is amplified and sequenced. Its genotype is B, serotype is adw and length is 3215 base.
The dominant genotype of HuNan is B, and the dominant serotype is adw.
Zhonghua shi yan he lin chuang bing du xue za zhi = Zhonghua shiyan he linchuang bingduxue zazhi = Chinese journal of experimental and clinical virology 07/2008; 22(3):205-7.
[Show abstract][Hide abstract] ABSTRACT: Detection HBV DNA among HBsAg negative children who have been vaccined at birth, in order to improve the evaluation of the indicator for HBV DNA infection.
Selection HBsAg negative children who have been vaccined at birth and then detection HBV DNA from sera using QIAamp Viral DNA Mini Kit, HBV DNA s region was obtained by nested PCR and sequencing.
12 of the 140 children were HBV DNA detected were positive and the infectious rate was 8.6% . No mutant of the 12 HBV DNA in "a" determinant.
To evaluate the effection of the prevention of HBV mother-to-child transmission, the standard method should be established. The detection of HBV DNA should be included in the future.
Zhonghua shi yan he lin chuang bing du xue za zhi = Zhonghua shiyan he linchuang bingduxue zazhi = Chinese journal of experimental and clinical virology 07/2008; 22(3):214-5.
[Show abstract][Hide abstract] ABSTRACT: DNA immunization has been used to induce either humoral or cellular immune responses against many antigens, including hepatitis C virus (HCV). In addition, DNA immunizations can be enhanced or modulated at the nucleotide level. Genetic immunizations were examined in BALB/c mice through the use of plasmids and chimeric DNA constructs encoding HCV core proteins and hepatitis B virus (HBV) precore (preC) regions. Plasmids encoding the truncated HCV core induced potent humoral and cellular responses to HCV; pcDNA3.0A-C154 produced a stronger antibody response than pcDNA3.0A-C191 (P < 0.01) and pcDNA3.0A-C69 (P < 0.05). HBV preC enhanced the humoral and cellular immune responses of BALB/c mice to HCV; however, pcDNA3.0A-C69preC resulted in a weak cytotoxic T lymphocyte (CTL) response. In addition, the humoral and cellular immune responses to HCV of groups immunized with pcDNA3.0A-C154preC and pcDNA3.0A-C191preC plasmids were higher than those of groups immunized with pcDNA3.0A-C154 and pcDNA3.0A-C191. In vivo CTL responses verified that mice immunized with preC core fused DNAs showed significantly high specific lysis compared with mice immunized with HCV cores only (P < 0.01). In our study, pcDNA3.0A-C154preC led to the highest immune response among all DNA constructs. Conclusion: DNA that encodes truncated HCV core proteins may lead to increased immune responses in vivo, and these responses may be enhanced by HBV preC.