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Inhibition of Hepatitis B Virus Replication and Expression in Vitro and in Vivo by the hammerhead Ribozymes Targeted Different Sites

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国际感染病学（电子版） 2012年第4期1卷 206-210页

作者： Wei Dai Rong Zhou Hong Yu Xiao-juan Li Department of Liver Disease Third People’s Hospital

Objective To develop an effective and specific medicine targeting hepatitis B virus(HBV) pregenome. Based on the identified accessible target sites for hammerhead ribozyme in our previous researches, a recombinant hepatitis delta virus(HDV) ribozyme was chosen and used to demonstrate the effective cleavage in vitro and in vivo. Methods Three hammerhead ribozymes for potential target sites(S, X and C genes) and co-expression plasmid(pTr-dB, pTdδ-dB, pTrX-dB and pTrC-dB) as well as four HDV-ribozyme chimera constructs with HBV(pTdXX, pTdXC, pTdSX and pTdSC) were severally chosen to validate the inhibition of the replication and expression of HBV. The co-expression plasmids(pTdδ and pTr-Db) in physiological saline were hydrodynamically injected to mice by tail vein. Results Compared with the group injected with pTr-dB in Huh-7 cell, hepatitis B surface antigen(HBsAg) was reduced by 31% in the group injected with pTdδ-dB, by 54%, 26%, 72% and 97% in the group injected with recombinant-ribozymes pTdSX, pTdSC, pTdXC and pTdXX, respectively. The inhibiting effects of endogenous ribozymes RzX and RzC on the HBsAg expression were 66% and 57%, respectively. Compared with the positive control, the amount of HBsAg was decreased in mice injected with pTdXX through tail vein by 88% and 96% on the second day and the third day, respectively. HBsAg was undetectable on the 6th day and could not primitively be detected on the 9th day in the sera from all mice. HBV DNA was not detected in the sera of BALB/c mice injected with pTdXX-dB, pTrX-dB or replicating-defective plasmid pHBV, while HBV DNA replication in control group could be detected on the 6th day. While HBcAg could not be detected in liver tissues of mice injected with plasmid pTdXX-dB on the 3rd day. Conclusions Encoding regions of HBV S, C and X gene were the effective cleavage sites for hammerhead ribozyme in vitro and in vivo, which provides basis for further construction of therapeutic recombinant HDV and the development

关键词： Hepatitis B virus(HBV) Pregenome RNA Hepatitis delta virus(HDV) hammerhead ribozyme

Effects of hammerhead ribozyme (RCP) on HBV P gene in vitro　

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Journal of Medical Colleges of PLA(China) 1996年第3期11卷 171-175页

作者：甘立霞王平朱锡华董燕麟侯云德

Based on the previous studies with the antisense RNA techique that the inhibitory effect on HBV gene expression came from the 5'-end and non-coding region of the P gene, a hammerhead ribozyme (RCP) was synthesized according to the ribozyme structural model published by Haseliff and Gerlach. This ribozyme targeted at the 5'-end of the P gene at 2360 site (GUC) on HBV ayw genome. It was found that RCP successfully cleaved its target RNA (340 nucleotide nt) and produced 2 specific degradation fragments. This finding provides a basis for the further study of its blocking effect on HBV gene expression and replication in vivo.

关键词： hammerhead ribozyme HBV cleavage, in vitro

极具未来科技风设计的自行车导航仪hammerhead

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工业设计 2013年第12期 50-50页

hammerhead是一款全新的自行车导航装置,无需分神即可安全地引导你到达目的地。这款hammerhead导航仪并没有内置GPS,它的工作原理可能是通过蓝牙与手机App连接,并通过控制彩色的LED灯和其他可视信息引导方向。它的应用程序配备了自行车... 详细信息

hammerhead是一款全新的自行车导航装置,无需分神即可安全地引导你到达目的地。这款hammerhead导航仪并没有内置GPS,它的工作原理可能是通过蓝牙与手机App连接,并通过控制彩色的LED灯和其他可视信息引导方向。它的应用程序配备了自行车专用地图来制定路线(支持苹果iOS和Android系统),无论路线长短都会确保是安全的,包括一些捷径和汽车无法通过的自行车道。你甚至可以根

关键词：自行车道内置这款导航仪 hammerhead

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基于单芯片的GPS接收机硬件设计

电子产品世界 2007年第6期14卷 104-105,107页

作者：胡睿黄光明华中师范大学物理学院电信系湖北省教育信息化研究中心

给出了基于hammerhead实现单芯片GPS接收机的硬件设计,给出射频PCB设计方案。

关键词： GPS接收机 hammerhead 射频