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| 1 | Optimization of competitively differentiated poiymerase chain reaction in detection of HBV basal core promoter mutation显示文摘AIM: To improve competitively differentiated polymerase chain reaction (CD-PCR) in detection of HBV basal core promoter mutation.METHODS: Recombinant plasmid of double point mutation A1762T/G1764A in basal core promoter of HBV constructed by site-directed mutagenesis was used as mutant control.To reveal the deficiency mechanism of CD-PCR, relationship between the circle number of PCR and the increased speed of products of each competitive primer was comparatively studied. Diversified amount of dNTPs and mutual primer of the competitive primers were tried to optimize CDPCR. Optimized CD-PCR was evaluated by detecting A1762T/G1764A mutation in recombinant plasmids and clinical sera from patients with HBV infection. RESULTS: The deficiency mechanism of CD-PCR was that the products of mismatched competitive primer grew fast when the amplification of matched primer entered into plateau stage, which led to decrease in or disappearance of the difference in the amount of their products. This phenomenon could be eliminated by reducing dNTPs to10 μmol/L and mutual primer to about 100 nmol/L. Optimized CD-PCR could detect both mutant and wild strain indepe ndent of the amount of templates and the number of PCRcycles. Its detection limit was 103 copies/mL, about 50 copies/reaction. About 10% of mutant DNAs among wild type DNAs could be detected. A1762T/G1764A mutant was detected in 41.8% (51/122) of patients with HBV infection, but not detected in controls with negative HBsAg. CONCLUSION: Optimized CD-PCR can detect mutation independent of the amount of initial templates and the number of PCR cycles. | Xiao-MouPeng LinGu Xue-JuanChen Jian-GuoLi Yang-SuHuang Zhi-LiangGao | 2005 | World Journal of Gastroenterology2005，11，23： | 8 |
| 2 | Yeast expression and DNA immunization of hepatitis B virus S gene with second-loop deletion of α determinant region显示文摘AIM: Immune escape mutations of HBV often occur in the dominant epitope, the second-loop of the a determinant of hepatitis B surface antigen (HBsAg). To let the hosts respond to the subdominant epitopes in HBsAg may be an effective way to decrease the prevalence of immune escape mutants. For this reason, a man-made clone of HBV S gene with the second-loop deletion was constructed. Its antigenicity was evaluated by yeast expression analysis and DNA immunization in mice. METHODS: HBV S gene with deleted second-loop, amino acids from 139 to 145, was generated using splicing by overlap extension. HBV deleted S gene was then cloned into the yeast expression vector pPIC9 and the mammalian expression vector pcDNA3 to generate pHB-SDY and pHB-SD, respectively. The complete S gene was cloned into the same vectors as controls. The deleted recombinant HBsAg expressed in yeasts was detected using Abbott IMx HBsAg test kits, enzyme-linked immunoadsorbent assay (ELISA) and immune dot blotting to evaluate its antigenicity in vitro. The anti-HBs responses to DNA immunization in BALB/c mice were detected using Abbott IMx AUSAB test kits to evaluate the antigenicity of that recombinant protein in vivo.RESULTS: Both deleted and complete HBsAg were successfully expressed in yeasts. They were intracellular expressions. The deleted HBsAg could not be detected by ELISA, in which the monoclonal anti-HBs against the determinant was used, but could be detected by Abbott IMx and immune dot blotting, in which multiple monoclonal antiHBs and polyclonal anti-HBs were used, respectively. The activity of the deleted HBsAg detected by Abbott IMx was much lower than that of complete HBsAg (the ratio of sample value/cut off value, 106+26.7 vs i 814.4+776.3, P<0.01,t=5.02). The anti-HBs response of pHB-SD to DNA immunization was lower than that of complete HBV S gene vector pHB (the positive rate 2/10 vs6/10, 4.56+3.52 mIU/mL vs27.60±17.3 mIU/mL, P=0.02, t=2.7). CONCLUSIONS: HBsAg with deleted second-loop of the α determinant still has antigenicity, and can also raise weak anti-HBs response in mice to DNA immunization, suggesting that it is possible to develop a subdominant vaccine for preventing infections of immune escape mutants of HBV. | HuiHu Xiao-MouPeng Yang-SuHuang LinGu Qi-FengXie Zhi-LiangGao | 2004 | World Journal of Gastroenterology2004，10，20： | 1 |
| 3 | Reversible Storage of Lithium in Silver‐Coated Three‐Dimensional Macroporous Silicon显示文摘 | YanYu LinGu ChangbaoZhu SusumuTsukimoto Peter A.van Aken JoachimMaier | 2010 | Adv Mater2010，，20： | 1 |
| 4 | Reversible Storage of Lithium in Silver‐Coated Three‐Dimensional Macroporous Silicon显示文摘 | YanYu LinGu ChangbaoZhu SusumuTsukimoto Peter A.van Aken JoachimMaier | 2010 | Adv Mater2010，，20： | 1 |
| 5 | The greedy and Delaunay triangulations are not bad in the average case 显示文摘 | LINGUS A | 1986 | Information Processing Letters1986，22，1： | 1 |
| 6 | The Greedy and Delaunay triangulations are not bad in the average case 显示文摘 | A Lingus | 1986 | Information Processing Letters1986，22，1： | 1 |
| 7 | Interleukin‐10 promoter polymorphisms are associated with the mode and sequel of HBeAg seroconversion in patients with chronic hepatitis B virus infection显示文摘 | Xiao MouPeng Yang SuHuang Hui HuiMa LinGu Qi FengXie Zhi LiangGao | 2006 | Liver International2006，，3： | 1 |
| 8 | ERROR ESTIMATION OF THE APPROXIMATION ALGORITHM FOR THE WINDY POSTMAN PROBLEM显示文摘If we restrict the postman to traversing each edge at most twice in the windypostman problem (WPP), we will get a new problem: 2WPP. An approximation algorithmhas been posed by M. Guan for the WPP. In the present paper, we improve the estimatederror given by M. Guan and show that we can estimate the error for the 2WPP by findinga minimum cost circulation. We also pose a new sufficient condition for the equivalencebetween WPP and 2WPP, which can be checked in polynomial time steps. | DU Lingu Shandong Textile Engineering College, Qingdao 266071, China | 1993 | Systems Science and Mathematical Sciences1993，6，2： | 0 |
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