ABSTRACT
Hepatitis C virus (HCV), one of the major pathogens of viral hepatitis, causes significant hazards in humans. Interferon treatment in combination with ribavirin is used as the first line clinical treatment for HCV infection. However, good response to this treatment has only been observed in few patients and repeated recurrence has also been reported frequently. Therefore, new antiviral agents and therapies are in urgent demand. Here, we report a newly constructed Escherichia coli RNase P based M1GS ribozyme that can specifically and efficiently target the core gene of HCV. The guide sequence (GS) of this M1IGS was designed according to the sequence of the core coding region of HCV genome. The GS was then covalently linked to the 3' terminus of M1 RNA, the catalytic subunit of RNase P from Escherichia coli. The specification of this sequence-specific ribozyme, M1GS, was then examined using an in vitro cleavage assay. The cytotoxicity and its activity in inhibition of HCV gene expression and viral proliferation were further studied in vivo. Our results show that the reconstructed M1GS ribozyme displayed obvious catalytic activity in cleaving target mRNAs fragment in vitro. Notable reduction in the expression of HCV core protein and a 1 000-fold reduction in viral growth were also observed in cultured HCV infected Huh7.5.1 cells expressing the functional M1GS ribozyme. This study demonstrated a direct evidence for the antiviral activity of the customized M1GS-HCV/C141 ribozyme, and thus provided a promising new strategy for clinical treatment of HCV infection.
Subject(s)
Antiviral Agents , Pharmacology , Escherichia coli , Genetics , Genetic Engineering , Hepacivirus , Genetics , Physiology , RNA, Catalytic , Genetics , Pharmacology , Genetics , Ribonuclease P , Genetics , Viral Core Proteins , GeneticsABSTRACT
AIM: To observe the treatment effectiveness of menopathy of excessive-cold syndrome by Modified Wenjing Decoction, and to explore the relationship among excessive-cold syndrome of menopathy and 5-HT,?-EP, and reproduetive hormones. METHODS: Sixty patients with menopathy of excess-cold syndrome were successively treated with Modified Wenjing Decoction for three menstrual cycles. The efficacy and the changes of serum 5-HT, Bendorphine(?-EP) and reproductive hormones before and after treatment were observed. RESULTS: 22 patients(36.7%) were short-term therapeutic effect, 28 patients(45.7%) were effective and 10 patients(17.6%) were ineffective. After treatment, the levels of serum FSH、LH、E_2、?-EP were increased significantly (P