Hepatitis: Cheng J

Cheng J. · No affiliation provided · Zhonghua Yi Xue Za Zhi. · Pubmed #17064585 No free full text.

This publication has no abstract.

Tsukamoto H, She H, Hazra S, Cheng J, Wang J. · USC-UCLA Research Center of ALPD and Cirrhosis, Department of Pathology, Keck School of Medicine, University of Southern California, Los Angeles, California, USA. · J Gastroenterol Hepatol. · Pubmed #18336651 No free full text.

Abstract: Alcoholic and non-alcoholic steatohepatitis (ASH and NASH) constitute two major types of chronic liver disease with worldwide prevalence and are histologically indistinguishable with shared pathogenetic mechanisms. More importantly, they have synergistic interactions for liver pathology. Comparative studies on ASH and NASH have been hampered by the use of different animal models with confounding variables, particularly those with extreme genetic, toxic, and malnutrition etiologies. The mouse intragastric model circumvents these problems and reproduces the natural course and etiological background of ASH and NASH. Further, our recent work reproduces a profound synergism between the two in the model. Intracellular accumulation of neural lipids is a hallmark biochemical feature of ASH and NASH. Although impaired lipid oxidation and export may contribute to this pathological change, enhanced lipogenic regulation is frequently encountered, as characterized by induction of lipogenic or adipogenic transcription factors (peroxisome proliferator-activated receptor [PPAR gamma], liver X receptor alpha[LXR alpha], sterol-regulatory element-binding protein-1c [SREBP-1c]). In contrast, we have recently defined transdifferentiation of hepatic stellate cells (HSC), a pivotal event in liver fibrogenesis, as an 'antilipogenic' or 'anti-adipogenic' phenomenon. Thus, there is an apparent paradox between hepatocytes and HSC in steatohepatitis in terms of the outcome of lipogenic regulation. Our recent work suggests that defective insulin signaling in activated HSC may be responsible for this paradox. Further, activated Wnt signaling is implicated in 'anti-adipogenic' stellate cell transdifferentiation in liver fibrogenesis.

Cheng J, Li L. · Gene Therapy Research Center, Institute of Infectious Diseases, 302nd Hospital of People's Liberation Army, Beijing 100039, China. · Zhonghua Gan Zang Bing Za Zhi. · Pubmed #12681078 No free full text.

This publication has no abstract.

Cheng J. · Gene Therapy Research Center, Institute of Infectious Diseases, The 302 Hospital of PLA, Beijing, China. · J Gastroenterol Hepatol. · Pubmed #12472961 No free full text.

This publication has no abstract.

Fan H, Zhao ZJ, Cheng J, Su XW, Wu QX, Shan YF. · Key Laboratory of Developmental Genes and Human Diseases, Ministry of Education, Southeast University, Nanjing, Jiangsu Province, China. · World J Gastroenterol. · Pubmed #19399937 links to  free full text

Abstract: AIM: To explore the relationship between DNA methyltransferase 1 (DNMT1) and hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) and its biological significance in primary HCC. METHODS: We carried out an immunohistochemical examination of DNMT1 in both HCC and paired non-neoplastic liver tissues from Chinese subjects. DNMT1 mRNA was further examined in HCC cell lines by real-time PCR. We inhibited DNMT1 using siRNA and detected the effect of depletion of DNMT1 on cell proliferation ability and cell apoptosis in the HCC cell line SMMC-7721. RESULTS: DNMT1 protein expression was increased in HCCs compared to histologically normal non-neoplastic liver tissues and the incidence of DNMT1 immunoreactivity in HCCs correlated significantly with poor tumor differentiation (P = 0.014). There were more cases with DNMT1 overexpression in HCC with HBV (42.85%) than in HCC without HBV (28.57%). However, no significant difference in DNMT1 expression was found in HBV-positive and HBV-negative cases in the Chinese HCC group. There was a trend that DNMT1 RNA expression increased more in HCC cell lines than in pericarcinoma cell lines and normal liver cell lines. In addition, we inhibited DNMT1 using siRNA in the SMMC-7721 HCC cell line and found depletion of DNMT1 suppressed cells growth independent of expression of proliferating cell nuclear antigen (PCNA), even in HCC cell lines where DNMT1 was stably decreased. CONCLUSION: The findings implied that DNMT1 plays a key role in HBV-related hepatocellular tumorigenesis. Depletion of DNMT1 mediates growth suppression in SMMC-7721 cells.

Xu DP, Liu Y, Cheng J, Li XD, Dai JZ, Li L, Liang ZL, Bai L, Zhong YW, Xu ZH, Ren XQ, Zhang LX. · Viral Hepatitis Research Laboratory, Institute of Infectious Diseases, 302 Hospital of PLA, Beijing 100039, China. · Zhonghua Gan Zang Bing Za Zhi. · Pubmed #18983768 No free full text.

Abstract: OBJECTIVES: To analyze HBV drug-resistant mutations against nucleos(t)ide analogues at 12 reported sites in 340 patients with chronic hepatitis B. METHODS: Serum HBV DNA was extracted and a nested PCR assay was employed for the reverse transcriptase (RT) gene amplification. Direct sequencing of PCR product was performed. The significance of detected mutations was analyzed in view of clinical data of the patients. RESULTS: Drug-resistant mutations were detected in 68 patients taking lamivudine (LAM), 10 taking adefovir (ADV), 8 taking entecavir, and 1 taking telbivudine (LdT). M204V and M204I were the most common LAM-resistant mutations. The former usually emerged with L180M while the latter often emerged alone. N236T +/- A181 substitution was the most frequently seen ADV-resistant mutation. ETV-resistant mutations occurred on the basis of LAM-resistant mutations and T184 change was the most common form. LdT-resistance was observed as M204I. Interestingly, these drug-resistant mutations were detected in a few patients who had not been treated with nucleos(t)ide analogues. CONCLUSION: Detection of HBV drug-resistant mutations at multiple sites of the viral RT gene is valuable for discovering and verifying drug resistance and thus is very helpful in planning anti-HBV therapy.

Zhang LF, Cheng J, Li JC, Chen LD, Liu W. · Department of Gastroenterology, First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China. · Zhonghua Gan Zang Bing Za Zhi. · Pubmed #18423152 No free full text.

Abstract: OBJECTIVE: To screen and clone hepatocyte protein interacting with hepatitis C virus NS5ATP4A protein for studying its biological functions. METHODS: Bait plasmids of hepatitis C virus NS5ATP4A were constructed. After verifying that hepatitis C virus NS5ATP4A protein could be steadily expressed in AH109 yeast strain, yeast-two hybrid assay was performed by mating AH109 with Y187 which pre-transformed with liver cDNA library plasmids pACT2, and the diploidy yeast cells were plated on quadruple dropout (QDO) medium and assayed for X-a-gal activity. Nineteen yeast colonies which grew on QDO and had a-gal activity were obtained, and then the library plasmids were extracted and sequenced. RESULTS: Seven genes were screened out and one of them was a formerly unknown gene. They were associated with RNA synthesis, protein translation, cell cycling and tumor immunity. CONCLUSION: NS5ATP4A binding proteins were successfully screened, which offers new clues for further studying the signal transduction pathway of NS5ATP4A and the pathogenic mechanism of HCV.

Wang JJ, Zhao P, Jin XY, Cheng J, Qing S, Ding N. · The 302 Hospital of PLA, Beijing 100039 China. · Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. · Pubmed #18414705 No free full text.

Abstract: OBJECTIVE: To found the subcellular location of the human gene 6 transactivated by nonstructural protein 5A of hepatitis C virus (NS5ATP6). METHODS: Green fluorescent protein (GFP) expression vector pEGFP- NS5ATP6 was established. The pEGFP- NS5ATP6 was transfected into HepG2 cells, and analyze the subcellular location of the proteins expressed by NS5ATP6 through Green fluorescent microscopy after 24 hours. RESULTS: The pEGFP- NS5ATP6 gene was successful cloned, NS5ATP6 can express protein in cells and subcellularly located in cell plasma. CONCLUSION: NS5ATP6 can express protein, and the protein expressed by NS5ATP6 subcellularly located in cell plasma.

Wang DQ, Guo J, Cheng J, Zhang JK, Zhao LF, Hong Y, Zhang LY. · Institute of Infectious Diseases, Beijing Ditan Hospital, Beijing 100011, China. · Zhonghua Gan Zang Bing Za Zhi. · Pubmed #18304421 No free full text.

Abstract: OBJECTIVE: To screen proteins in leukocytes interacting with PS1TP2 by yeast-two hybrid and to view their subcellular localization in HepG2 cells. METHODS: The function and structure of PS1TP2 were studied by bioinformatic analysis. PS1TP2 gene was amplified and cloned into plasmid pET32a (+) and pGBKT7 to construct recombinant expression vectors pET32a (+)-PS1TP2 and pGBKT7-PS1TP2. They were transduced into E. coli Rosetta strain and yeast AH109. The transformed yeast mated with yeast Y187 containing leukocyte cDNA library plasmid in a 2xYPDA medium. Diploid yeast cells were plated on a synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) for selecting twice and then screening. Then a green fluorescent protein (GFP) expression vector pEGFP-C1-PS1TP2 was established, transduced into HepG2, and its subcellular localization was studied by fluorescence microscopy and confocal microscopy. RESULTS: Bioinformatic analysis showed that the PS1TP2 gene was located at 6q24.1, the protein was unstable and the aliphatic index was very high. After transformation of the E. coli and yeast AH109, the expression protein showed: (1) the molecular weight of the expressed product was about 41000 Da, and (2) PS1TP2 existed within the cells. Diploid yeast cells were plated on the synthetic dropout nutrient medium containing X-a-gal for selecting twice and then screening. Twenty-six colonies from blue colonies were sequenced, pEGFP-C1-PS1TP2 was successfully expressed in the HepG2 cells, and PS1TP2 was located in the cell plasma. CONCLUSION: A prokaryotic expression vector pET32a(+)-PS1TP2 was constructed successfully and the PS1TP2 was successfully expressed in the yeast system. Genes of PS1TP2 interact with leukocyte proteins. These results bring some new clues for studying the biological functions of HBV.

Shi L, Zhang SL, Li K, Hong Y, Wang Q, Li Y, Guo J, Fan WH, Zhang L, Cheng J. · The First Hospital of Xi'an Jiaotong University, Xi'an, China. · Cancer Lett. · Pubmed #18068894 No free full text.

Abstract: Non-structural protein 5A (NS5A) appears to interact with a variety of cellular proteins and play an important role in mediating cell growth, cellular signaling pathways and pathogenesis of hepatitis C virus (HCV). NS5ATP9 was identified as a NS5A trans-activated protein in suppression subtractive hybridization (SSH), and the regulation was confirmed by luciferase reporter assay and quantitative real time PCR (qRT-PCR). A minimal promoter region contained within the 211bp (nucleotides -161 to +50bp) immediately upstream of the transcription initiation site. NS5ATP9 is a NS5A up-regulation gene which may play a role in the pathogenesis of HCV-associated hepatocellular carcinoma.

Wang Z, Hou J, Zeng G, Wen S, Tanaka Y, Cheng J, Kurbanov F, Wang L, Jiang J, Naoumov NV, Mizokami M, Qi Y. · State Key Laboratory of Virology, College of Life Science, Wuhan University, Wuhan, China. · J Viral Hepat. · Pubmed #17501764 No free full text.

Abstract: Genetic diversity within the same hepatitis B virus (HBV) genotype indicates the presence of several subgenotypes. We have found that genotype C is the most common in China, and this study aimed to determine the geographical distribution and characteristics of HBV-C subgenotypes in the country. A cohort of 534 patients with chronic HBV genotype C infection, collected across China, was analysed by nucleotide sequencing or polymerase chain reaction-restriction fragment length polymorphism. HBV-C1/Cs (n = 112, 21%) and HBV-C2/Ce (n = 397, 74%) were the most common HBV-C subgenotypes and showed different geographical distribution in China. No significant differences were found between patients infected with HBV-C1 and HBV-C2 when comparing liver function tests, hepatitis B e antigen positive rate and clinical manifestations. We identified two other types of HBV-C provisionally designated as HBV-CD1 and HBV-CD2, which have particular virological features and clustered in one geographic area. These two types of C/D hybrids have emerged through recombination with genotype D and encode serotype ayw2 hepatitis B surface antigen. In conclusion, there are at least four subtypes of HBV genotype C: subgenotypes C1, C2 and two types of C/D recombinants CD1 and CD2 in China, which have a distinct geographic distribution. Whether HBV-C subgenotypes differ in their impact on liver disease progression requires prospective studies.

Zhang JK, Zhao LF, Cheng J, Guo J, Wang DQ, Hong Y, Mao Y. · Department of Gastroenterology, The First Hospital of Shanxi Medical University, Taiyuan 030001, Shanxi Province, China. · World J Gastroenterol. · Pubmed #17461456 links to  free full text

Abstract: AIM: To clone and identify human genes transactivated by PS1TP5 by constructing a cDNA subtractive library with suppression subtractive hybridization (SSH) technique. METHODS: SSH and bioinformatics techniques were used for screening and cloning of the target genes transactivated by PS1TP5 protein. The mRNA was isolated from HepG2 cells transfected with pcDNA3.1(-)-myc-his(A)-PS1TP5 and pcDNA3.1(-)-myc-his(A) empty vector, respectively, and SSH technique was employed to analyze the differentially expressed DNA sequence between the two groups. After digestion with restriction enzyme Rsa I, small size cDNAs were obtained. Then tester cDNA was divided into two groups and ligated to the specific adaptor 1 and adaptor 2, respectively. The tester cDNA was hybridized with driver cDNA twice and subjected to nested PCR for two times, and then subcloned into T/A plasmid vectors to set up the subtractive library. Amplification of the library was carried out with E. coli strain DH5alpha. The cDNA was sequenced and analyzed in GenBank with Vector NTI 9.1 and NCBI BLAST software after PCR amplification. RESULTS: The subtractive library of genes transactivated by PS1TP5 was constructed successfully. The amplified library contained 90 positive clones. Colony PCR showed that 70 clones contained 200-1000-bp inserts. Sequence analysis was performed in 30 clones randomly, and the full-length sequences were obtained by bioinformatics technique. Altogether 24 coding sequences were obtained, which consisted of 23 known and 1 unknown. One novel gene with unknown functions was found and named as PS1TP5TP1 after being electronically spliced, and deposited in GenBank (accession number: DQ487761). CONCLUSION: PS1TP5 is closely correlated with immunoregulation, carbohydrate metabolism, signal transduction, formation mechanism of hepatic fibrosis, and occurrence and development of tumor. Understanding PS1TP5 transactive proteins may help to bring some new clues for further studying the biological functions of pre-S1 protein.

Bai GQ, Yue YF, Zhang SL, Cheng J, Liu Y, Li SH, Zhang XE. · Department of Obstetrics and Gynecology, First Hospital, Xi'an Jiaotong University, Xi'an 710061, China. · Zhonghua Fu Chan Ke Za Zhi. · Pubmed #17442177 No free full text.

Abstract: OBJECTIVE: To screen differentially expressed genes in placentas with hepatitis B virus (HBV) infection and to discuss the molecular mechanism of HBV intrauterine infection. METHODS: Thirty placenta tissue specimens from HBsAg and HBV DNA positive pregnant women were used as the study group and 30 placenta tissue specimens from normal pregnant women with HBsAg and HBV DNA negativity were served as the control group. The suppression subtractive hybridization (SSH) technique was used. Total RNAs of placenta tissue of the study group were mixed as the tester, and total RNAs of placenta tissue of the control group were mixed as the driver. A subtractive cDNA library was constructed by PCR-selective cDNA subtraction systems. Amplifications of the library were carried out with E. coli strain DH5alpha by reverse spot hybridization. RT-PCR confirmed that phosphatidylinositol 3-kinase (PI3K) was up-regulated in placenta tissue with HBV infection. RESULTS: Colony PCR showed that the clones contained 200 - 1000 bp inserts. Thirty five clones were confirmed by reverse spot hybridization and analyzed by sequencing and bioinformatics. Thirty three known genes and 2 genes with unknown function were obtained. RT-PCR preliminarily confirmed that PI3K gene was up-regulated in HBV infected placenta. CONCLUSIONS: The differentially expressed genes in placentas with hepatitis B virus (HBV) infection using SSH technique has been screened out successfully. These differentially expressed genes encoding proteins participating in cell vital metabolism and malformation, and signal conduction-antiapoptosis pathway. This finding brings some new clues for studying the mechanisms of HBV intrauterine infection.

Cheng YQ, Wang L, Cheng J, Liu Y, Xu DP, Zhong YW, Qu JH, Tian JK, Dai JZ, Li XD. · Viral Hepatitis Research Center, The No.302 Hospital of People's Liberation Army, Beijing 100039, China. · Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. · Pubmed #17429534 No free full text.

Abstract: OBJECTIVE: To screen proteins interacting with HCV NS4A protein in leukocytes by yeast-double hybridization. METHODS: The bait plasmid pGBKT7-NS4A was transformed into yeast AH109 was transformed, and the expressing of the fusion protein was identified by SDS-page. The transformed yeast was mated with yeast Y187 containing leukocytes cDNA library plasmid in 2xYPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) and synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing x-alpha-gal for selecting two times and screening. After extracting and sequencing of plasmid from blue colonies, analysis was conducted by bioinformatics. And, the gene encoding the interesting protein was cloned, and back-cross was performed. RESULTS: Forty-five colonies were sequenced, among them, 29 colonies were human calcium modulating cyclophilin ligand (CAML). The gene encoding CAML was cloned, and the interaction between NS4A and CAML was ensured. CONCLUSION: Seven kinds of proteins interacting with NS4A in leukocytes were successfully screened and the results brought some new clues for studying the pathogenesis of HCV.

Cheng J. · Institute of Infectious Diseases, Beijing Ditan Hospital, Beijing 100011, China. · Zhonghua Gan Zang Bing Za Zhi. · Pubmed #17362640 No free full text.

This publication has no abstract.

Li ZQ, Ma YJ, Cheng J. · Institute of Infectious Diseases, Beijing Ditan Hospital, Beijing 100011, China. · Zhonghua Gan Zang Bing Za Zhi. · Pubmed #17362635 No free full text.

Abstract: OBJECTIVES: To screen proteins in hepatocytes interacting with hepatitis B virus surface antigen middle protein (MHBs) with yeast-two hybrid technique for studying the biological functions of MHBs. METHODS: Polymerase chain reaction (PCR) was used to amplify the gene of MHBs from the plasmid A7 containing the whole fragment of adr subtype of HBV and the PCR product was cloned into pGEM-T vector and then evaluated by sequencing. The gene of MHBs was cut by EcoRI and BamH I from pGEM-T vector and then cloned into the yeast expression plasmid pGBKT7. MHBs bait plasmid was constructed by ligating MHBs gene with yeast expression vector pGBKT7 with yeast-two hybrid system 3 and then was transformed into yeast AH109 (a type). The transformed yeast cells were mated with yeast Y187 (alpha type) containing liver cDNA library plasmid in 2 x YPDA medium. Diploid yeast cells were plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) and synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing X- alpha -gal for selecting and screening. After extracting and sequencing the plasmid from true positive blue colonies, the results were analyzed by bioinformatics. RESULTS: A pGBKT7- MHBs yeast expressed vector was successfully constructed. Two colonies were sequenced. One colony was Homo sapiens aldolase B fructose-bisphosphate, the other was a new gene with unknown function, which was named MHBs-binding protein 1. CONCLUSION: MHBs gene was successfully cloned. Two genes of MHBs interacting proteins in hepatocytes were obtained by yeast-two hybrid system 3. Our results brought some new clues for studying the biological functions of MHBs and the mechanisms of HBV carcinogenesis.

Zhang JK, Zhao LF, Cheng J, Guo J, Lun YZ, Hong Y. · Department of Gastroenterology, First Hospital of Shanxi Medical University, Taiyuan 030001, China. · Chin Med J (Engl). · Pubmed #17134587 links to  free full text

Abstract: BACKGROUND: The hepatitis B virus (HBV) genome includes S, C, P and X regions. The S region is divided into four subregions of pre-pre-S, pre-S1, pre-S2 and S. PS1TP5 (human gene 5 transactivated by pre-S1 protein of HBV) is a novel target gene transactivated by the pre-S1 protein that has been screened with a suppression subtractive hybridization technique in our laboratory (GenBank accession: AY427953). In order to investigate the biological function of the PS1TP5 protein, we performed a yeast two-hybrid system 3 to screen proteins from a human leukocyte cDNA library interacting with the PS1TP5 protein. METHODS: The reverse transcription polymerase chain reaction (RT-PCR) was performed to amplify the gene of PS1TP5 from the mRNA of HepG2 cells and the gene was then cloned into the pGEM-T vector. After being sequenced and analyzed with Vector NTI 9.1 and NCBI BLAST software, the target gene of PS1TP5 was cut from the pGEM-T vector and cloned into a yeast expression plasmid pGBKT7, then "bait" plasmid pGBKT7-PS1TP5 was transformed into the yeast strain AH109. The yeast protein was isolated and analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting hybridization. After expression of the pGBKT7-PS1TP5 fusion protein in the AH109 yeast strain was accomplished, a yeast two-hybrid screening was performed by mating AH109 with Y187 containing a leukocyte cDNA library plasmid. The mated yeast was plated on quadruple dropout medium and assayed for alpha-gal activity. The interaction between the PS1TP5 protein and the proteins obtained from positive colonies was further confirmed by repeating the yeast two-hybrid screen. After extracting and sequencing of plasmids from blue colonies we carried out a bioinformatic analysis. RESULTS: Forty true positive colonies were selected and sequenced, full length sequences were obtained and we searched for homologous DNA sequences from GenBank. Among the 40 positive colonies, 23 coding genes with known functions were obtained, including Homo sapien leukocyte adhesion protein p150, 95, interleukin 2 receptor gamma chain, PALM2-AKAP2 protein (PALM2-AKAP2), eukaryotic translation initiation factor 4A, beta-2-microglobin, solute carrier family 9 (sodium/hydrogen exchanger), calreticulin, asialoglycoprotein receptor 1 (ASGR1), MHC class II lymphocyte antigen, cytochrome c oxidase subunit 1, lymphocyte antigen 86 (LY86) and lymphocyte cytosolic protein 1. One novel gene with unknown function was found and named as PS1TP5BP1. After being electronically spliced, it was deposited in GenBank (accession number: DQ471327). CONCLUSIONS: Genes of proteins interacting with PS1TP5 were successfully screened from leukocyte cDNA library. These results suggested that PS1TP5 was closely correlated with immunoregulation, carbohydrate metabolism, signal transduction, the formation of hepatic fibrosis and initiation and development of tumors and also brought some new clues for further studying the biological functions of the pre-S1 protein.

Lun YZ, Cheng J, Zhong YW, Zhao BC. · Department of Biochemistry and Molecular Biology, Dalian Medical University, Dalian 110027, P.R. China. · Acta Virol. · Pubmed #17131939 No free full text.

Abstract: Expression of single-chain variable fragment (scFv) antibodies on the surface of bacteriophage is widely used to prepare antibodies with pre-defined specificities. A phage antibody library containing the gene for scFv antibody against Hepatitis B virus surface antigen (HBsAg) was panned with HBsAg immobilized on microtiter plate wells. After five rounds of panning 30 phage clones specific to HBsAg were obtained and one selected clone was sequenced. It was found to consist of 789 bp and its amino acid sequence and specifically detected the respective antigen in the patients but not in healthy persons.

Zhong YW, Cheng J, Qu JH, Zhang LY, Guo J, Li XD, Xu DP. · Viral Hepatitis Research Center, Institute of Infectious Diseases, The No.302 Hospital of The People's Liberation Army, Beijing 100039, China. Corresponding author: ZHONG Yan-wei, E-mail: , Tel: 010-66933392. · Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. · Pubmed #17086292 No free full text.

Abstract: BACKGROUND: To construct a subtractive cDNA library of target genes down-regulated in human hepatocarcinoma cell line HepG2 cells treated with IFNB, and clone genes of the down-regulation by IFNB using suppression subtractive hybridization (SSH) technology and bioinformatics techniques. METHODS: The mRNA was isolated from HepG2 cells induced by recombinant interferon-B and 0.9 percent sodium chloride, respectively, then cDNA was synthesized. After restriction enzyme Rsa I digestion, small sizes cDNAs were obtained. Then tester cDNA was divided into two portions and each was ligated to the specific cDNA adaptor 1 and adaptor 2 respectively. After tester cDNA was hybridized with driver cDNA twice and underwent two times of nested PCR, the DNA fragment was subcloned into T/A plasmid vectors to set up the subtractive library. Amplification of the library was carried out with E.coli strain DH5a. The cDNA was sequenced and analyzed in GenBank with Blast search after PCR. RESULTS: The subtractive library of genes down-regulation in HepG2 cells treated with recombination interferon-B was constructed successfully. The amplified library contained 58 positive clones. Colony PCR and sequence analysis was performed in 35 clones randomly, and the full length sequences were obtained with bioinformatics method. Altogether 12 coding sequences were obtained. CONCLUSION: A subtractive cDNA library of genes down-regulation in HepG2 cells treated with IFNB using SSH technique was constructed successfully, which brings some new clues for studying the regulation mechenism of IFNB in liver cells.

Liu Y, Bai GQ, Cheng J, Yang Q, Zhang LY, Ji D, Wang JJ. · Viral Hepatitis Research Center, Institute of Infectious Diseases, The No.302 Hospital of PLA, Beijing 100039, China. · Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. · Pubmed #17086291 No free full text.

Abstract: BACKGROUND: To construct a subtractive cDNA library of genes transactivated by NS4A protein of hepatitis C virus with suppression subtractive hybridization technique (SSH). METHODS: The mRNA was isolated from Hep G2 cells transfected pcDNA3.1(-)-NS4A and pcDNA3.1(-) empty vector, respectively, then the cDNA was synthesized. SSH method was employed to analyze the differentially expressed RNA sequence between the two groups. The twice enriched PCR products were subcloned into T/A vectors to set up the subtractive library. Amplification of the library was carried out with E.coli strain JM109. The cDNA was sequenced and analyzed in comparison with those in GenBank with Blast search after PCR. RESULTS: The amplified library contained 36 positive clones. Colony PCR showed that 36 clones contained 200-1000 bp inserts. Sequence analysis was performed in 25 clones, and the full length sequences were obtained with bioinformatics method. Altogether 20 kinds of coding sequences were achieved, which consisted of 18 kinds of known and 2 kinds of unknown ones. The obtained sequences may be target genes transactivated by NS4A protein of HCV, among which some genes coding for proteins involved in cell cycle regulation, cell apoptosis, signal transduction pathway and tumour development. CONCLUSION: A subtractive library of genes transactivated by NS4A protein of HCV was constructed successfully, which brought some new clues for studying the biological functions and pathogenesis of the viral proteins.

Wei HS, Huang YB, Song SJ, Dong QM, Li GL, Cheng J. · Institute of Infectious Diseases, Beijing Ditan Hospital, Beijing 100011, China. · Zhonghua Gan Zang Bing Za Zhi. · Pubmed #16867283 No free full text.

This publication has no abstract.

Lin SM, Cheng J, Lu YY, Zhang SL, Yang Q, Chen TY, Liu M, Wang L. · Department of Infectious Diseases, The First Affiliated Hospital of Medical College of Xi'an Jiaotong University, Xi'an 710061, Shaanxi Province, China. · World J Gastroenterol. · Pubmed #16534844 links to  free full text

Abstract: AIM: To investigate the biological function of HBcAg in pathogenesis of HBV replication in peripheral blood mononuclear cells (PBMCs). METHODS: HBcAg region was amplified by polymerase chain reaction (PCR) and HBV HBcAg bait plasmid pGBKT7-HBcAg was constructed by routine molecular biological methods. Then the recombinant plasmid DNA was transformed into yeast AH109. After the HBV core protein was expressed in AH109 yeast strains (Western blot analysis), yeast-two hybrid screening was performed by mating AH109 with Y187 containing leukocyte cDNA library plasmid. Diploid yeast cells were plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) (QDO) and synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) (TDO). The second screening was performed with the LacZ report gene ( yeast cells were grown in QDO medium containing X-alpha-gal). The interaction between HBV core protein and the protein obtained from positive colonies was further confirmed by repeating yeast-two hybrid. After plasmid DNA was extracted from blue colonies and sequenced, the results were analyzed by bioinformatic methods. RESULTS: Eighteen colonies were obtained and sequenced, including hypermethylated in cancer 2 (3 colones), eukaryotic translation elongation factor 2 (2 colones), acetyl-coenzyme A synthetase 3 (1 colone), DNA polymerase gamma (1 colone), putative translation initiation factor (1 colone), chemokine (C-C motif) receptor 5 (1 colone), mitochondrial ribosomal protein L41 (1 colone), kyot binding protein genes (1 colone), RanBPM (1 colone), HBeAg-binding protein 3 (1 colone), programmed cell death 2 (1 colone). Four new genes with unknown function were identified. CONCLUSION: Successful cloning of genes of HBV core protein interacting proteins in leukocytes may provide some new clues for studying the biological functions of HBV core protein.

Wang L, Cheng J, Li K, Hong Y. · Center for Viral Hepatitis Research, Institute of Infectious Diseases, 302 Hospital of PLA, Beijing 100039, China. · Zhonghua Gan Zang Bing Za Zhi. · Pubmed #16494772 No free full text.

Abstract: OBJECTIVE: To clarify the expression and regulation mechanism of the new target gene (human hepatitis C virus binding protein 6, Hcbp6) interaction with the core protein of hepatitis C virus (HCV). METHODS: Referring to the prediction online of the promoter region, 3256 base pairs (bp) upstream and downstream of the translation start site were selected as 5 putative promoter sequences which were amplified from the hepatoblastoma cell line-HepG2 cell genomic DNA by polymerase chain reaction (PCR). The amplified products were cloned into a pCAT3 vector. The HepG2 and NIH3T3 cell lines were transfected by pCAT3-Hcbp6-promoters. The CAT activity was detected using an enzyme-linked immunoassay (ELISA) kit. RESULTS: We found that 2 kinds of pCAT3-Hcbp6-promoters (1066 and 240) could direct the reporter gene expression. The expression of CAT was 3.1 times and 6.4 times higher than that of the pCAT3-basic control vector. CONCLUSION: These results indicated that pCAT3-Hcbp6-promoters have promoter activity.

Wei HS, Dong QM, Zhuang H, Song SJ, Liu ZY, Cheng J. · Research Center of Virology, Beijing Ditan Hospital, Beijing 100011, China. · Zhonghua Gan Zang Bing Za Zhi. · Pubmed #16420769 No free full text.

This publication has no abstract.

Cheng J. · Institute of Infectious Diseases, Beijing Ditan Hospital, Beijing 100011, China. · Zhonghua Gan Zang Bing Za Zhi. · Pubmed #16420768 No free full text.

This publication has no abstract.