Chih-Hao Liu

National Taiwan University, T’ai-pei, Taipei, Taiwan

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Publications (5)11.86 Total impact

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    ABSTRACT: ETHNOPHARMACOLOGICAL RELEVANCE: Ching-fang-pai-tu-san (CFPTS) is a Chinese herbal decoction that is used as a cure for the common cold, fever, headache, and poor circulation. However, no previous studies have investigated the mode of action of CFPTS against influenza virus infections. To investigate the antiviral mechanism of CFPTS, we examined viral entry, transcription, translation, viral glycoprotein hemagglutinin (HA) transport, and budding of the influenza virus. MATERIALS AND METHODS: The antiviral activity of nontoxic concentrations of CFPTS against influenza virus A/WSN/33 was examined by assaying (neutralization assay) its inhibition of the virus-induced cytopathic effects. The mode of CFPTS action was first examined with a time-of-addition assay of synchronized infections, followed by monitoring HA transport by immunofluorescence microscopy. Viral endocytosis was evaluated with attachment and penetration assays. The inhibition of viral replication was measured by quantitative real-time PCR, immunoblotting, and immunofluorescence microscopy. We also performed assays related to the inhibition of viral entry, such as neuraminidase activity and hemagglutinin activity assays. RESULTS: Based on the inhibition of the virus-induced cytopathic effect in Madin-Darby canine kidney cells, the EC(50) of CFPTS was about 1.44±0.22mg/mL against influenza virus A/WSN/33. CFPTS displayed a broad spectrum of inhibitory activities against different strains of influenza A virus, as well as some enteroviruses. However, this extract proved less effective against clinical oseltamivir-resistant strains and influenza B viruses. CFPTS did not suppress viral RNA or protein synthesis. According to a time-of-addition assay, the antiviral mechanism of CFPTS may involve viral budding or intracellular viral glycoprotein transport. A plaque reduction assay showed that CFPTS reduced both the plaque size and plaque quantity. The intracellular transport of viral glycoprotein hemagglutinin was blocked by CFPTS by immunofluorescence microscopic analysis. Thus, it is possible that the antiviral mechanism of CFPTS might inhibit the assembly of progeny virions and/or their subsequent release. CONCLUSIONS: Our results give scientific support to the use of CFPTS in the treatment of influenza virus infections. CFPTS has potential utility in the management of seasonal pandemics of influenza virus infections, like other clinically available drugs.
    Journal of ethnopharmacology 10/2012; 144(3). DOI:10.1016/j.jep.2012.09.035 · 2.94 Impact Factor
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    ABSTRACT: We developed a biomolecular layer with highly specific binding that enables determination of low concentrations of 3,4-methylenedioxymethylamphetamine (MDMA) by the immobilization of antibodies. This study examined the interactions of anti-MDMA antibodies with MDMA using a microcantilever-based biosensor. The detection platform is based on the immobilization of affinity monoclonal antibodies onto a sensing surface using a cysteamine-based self-assembled monolayer. Anti-MDMA antibodies conjugated with various concentrations of MDMA were bound to the sensing surface. The intermolecular interaction was observed by monitoring the simultaneous signals (bending deflection and resonant frequency shift) of microcantilevers in phosphate buffered saline (PBS). We demonstrated that anti-MDMA antibodies can respond specifically to their natural ligand, MDMA. Results revealed that the microcantilever-based biosensor can be used to characterize MDMA response profiles of anti-MDMA and provide rich data for immunoassays. In addition to providing a clearer understanding of the biological mechanisms of protein binding, such a biosensor also holds great potential in numerous other fields, such as drug detection, biomedicine, and environmental protection.
    Sensors and Actuators A Physical 08/2012; 182:163–167. DOI:10.1016/j.sna.2012.05.036 · 1.94 Impact Factor
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    ABSTRACT: Ma-xing-shi-gan-tang (MXSGT, aka maxing shigan powder), a Chinese herbal decoction, has been used for the treatment of the common cold, fever, and influenza virus infections. However, the underlying mechanisms of its activity against the influenza virus are not fully understood. In this study, we examined the antiviral effects of MXSGT in influenza-virus-infected MDCK cells and their underlying mechanisms, including the damage of the viral surface ultrastructure and the consequent inhibition of viral entry.
    Journal of ethnopharmacology 06/2012; 143(1):57-67. DOI:10.1016/j.jep.2012.05.061 · 2.94 Impact Factor
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    ABSTRACT: Atomic force microscopy (AFM) is a vital instrument in nanobiotechnology. In this study, we developed a method that enables AFM to simultaneously measure specific unbinding force and map the viral glycoprotein at the single virus particle level. The average diameter of virus particles from AFM images and the specificity between the viral surface antigen and antibody probe were integrated to design a three-stage method that sets the measuring area to a single virus particle before obtaining the force measurements, where the influenza virus was used as the object of measurements. Based on the purposed method and performed analysis, several findings can be derived from the results. The mean unbinding force of a single virus particle can be quantified, and no significant difference exists in this value among virus particles. Furthermore, the repeatability of the proposed method is demonstrated. The force mapping images reveal that the distributions of surface viral antigens recognized by antibody probe were dispersed on the whole surface of individual virus particles under the proposed method and experimental criteria; meanwhile, the binding probabilities are similar among particles. This approach can be easily applied to most AFM systems without specific components or configurations. These results help understand the force-based analysis at the single virus particle level, and therefore, can reinforce the capability of AFM to investigate a specific type of viral surface protein and its distributions.
    Biochemical and Biophysical Research Communications 11/2011; 417(1):109-15. DOI:10.1016/j.bbrc.2011.11.065 · 2.28 Impact Factor
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    ABSTRACT: The product and direct role of the rssC gene of Serratia marcescens is unknown. For unraveling the role of the rssC gene, atomic force microscopy has been used to identify the surfaces of intact S. marcescens wild-type CH-1 cells and rssC mutant CH-1ΔC cells. The detailed surface topographies were directly visualized, and quantitative measurements of the physical properties of the membrane structures were provided. CH-1 and CH-1ΔC cells were observed before and after treatment with lysozyme, and their topography-related parameters, e.g., a valley-to-peak distance, mean height, surface roughness, and surface root-mean-square values, were defined and compared. The data obtained suggest that the cellular surface topography of mutant CH-1ΔC becomes rougher and more precipitous than that of wild-type CH-1 cells. Moreover, it was found that, compared with native wild-type CH-1, the cellular surface topography of lysozyme-treated CH-1 was not changed profoundly. The product of the rssC gene is thus predicted to be mainly responsible for fatty-acid biosynthesis of the S. marcescens outer membrane. This study represents the first direct observation of the structural changes in membranes of bacterial mutant cells and offers a new prospect for predicting gene expression in bacterial cells.
    Microscopy and Microanalysis 10/2010; 16(6):755-63. DOI:10.1017/S1431927610093943 · 1.76 Impact Factor