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1.
Chinese Journal of Experimental and Clinical Virology ; (6): 150-152, 2007.
Article in Chinese | WPRIM | ID: wpr-248819

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate feasibility of inhibiting coxsackievirus B3 (CVB3) infection at cellular, protein and gene levels by using small interfering RNA (siRNA).</p><p><b>METHODS</b>Antiviral activities of siRNAs were evaluated by observing cytopathic effect (CPE), using plaque reduction Western blotting assays and RT-PCR.</p><p><b>RESULTS</b>Eight siRNAs were synthesized, among them, SiRNA-2, SiRNA-3, SiRNA-6 and SiRNA-7 which were targeted against sequences located in 2B, VP4, 2A and 3C section of CVB3 genome, were designed to have different effect of inhibiting CVB3 infection in vitro. SiRNA-2 showed the best protective effect, 95 percent inhibition of CVB3 cytopathic effect and plaque forming effect was observed at 0.0001 MOI, viral protein synthesis and replication were inhibited. SiRNA-2 showed 30 percent inhibition of virus at 0.1 MOI, 70 percent inhibition at 0.01 MOI, 88 percent inhibition at 0.001 MOI, and 99 percent inhibition at 0.0001 MOI 48 hours after CVB3 infection.</p><p><b>CONCLUSION</b>SiRNA could effectively inhibit CVB3 infection in vitro, siRNA-2, which is targeted against sequence in 2B section of CVB3 genome, seemed to be the best one among those synthesized in this study.</p>


Subject(s)
Humans , Coxsackievirus Infections , Therapeutics , Virology , Cytopathogenic Effect, Viral , Enterovirus , Genetics , Physiology , HeLa Cells , RNA Interference , RNA, Small Interfering , Genetics , Therapeutic Uses , Virus Replication
2.
Chinese Journal of Virology ; (6): 276-281, 2007.
Article in Chinese | WPRIM | ID: wpr-334897

ABSTRACT

To study the inhibitory effect and function characteristics of small interfering RNA (siRNA) on cosxackievirus B3(CVB3) infection by RNA interference technique, siRNA-2B against the viral 2B region was synthesized and transfected into HeLa cell, which was then infected with CVB3. The efficiency of siRNA transfection was examined by FCM, the cell toxicity of siRNA-2B by MTT, and the antiviral ability of siRNA-2B by cytopathic effect (CPE), plaque reduction assay and RT-PCR. The results showed that siRNA-2B could be transfected efficiently into HeLa cell and lasted at least 48h. High concentration of siRNA-2B didn't show any sign of toxicity to cells. siRNA-2B exhibited a significant protective effect on cell viability by specific inhibition of viral replication. It showed a close relationship between the concentrations of siRNA-2B and the antiviral effects. siRNA-2B led to dramatical reduction of viral titers in supernatant of cell culture and weakened the reinfection ability of the virus. These data proposed that siRNA-2B, targeting 2B protein, can effectively inhibit CVB3 infection in HeLa cell and exhibits its transfection efficiency, viral inhibition specificity and adose-dependant manner, suggesting its potential role in prevention and treatement of CVB3 infection.


Subject(s)
Humans , Enterovirus , Genetics , Green Fluorescent Proteins , Genetics , Metabolism , HeLa Cells , Microscopy, Fluorescence , Plasmids , Genetics , RNA, Small Interfering , Genetics , Recombinant Fusion Proteins , Genetics , Metabolism , Transfection , Viral Nonstructural Proteins , Genetics , Virus Replication , Genetics , Physiology
3.
China Journal of Chinese Materia Medica ; (24): 269-279, 2006.
Article in Chinese | WPRIM | ID: wpr-350961

ABSTRACT

In this article, the research of chemical constituents and bioactivities in recent ten years has been reviewed of plants from the 12 genera in Flacourtiaceae related to the medicinal resources in China. The research in China about the plants from Flacourtiaceae was done very little, but many literatures have been reported abroad. The plants from Flacourtiaceae mostly contain the constituents such as aromatic glucosides, lignanoid glucosides, diterpenoids and cyclopentenoid cyanohydrin glucosides et al. These compounds or plant extracts mainly show antibacterial, antiviral, antitumor, hypolipidemic and hypoglycemic activities. The research of plants from Carrierea, Itoa and Bennettiodendron of Flacourtiaceae in China is still blank. The systemic research about chemical constituents and bioactivities of plants from these genera will play important roles in the discovery of novel natural products and active constituents, and provide valuable reference for the classifying of plants from these genera.


Subject(s)
Animals , Humans , Anti-Bacterial Agents , Pharmacology , Antineoplastic Agents, Phytogenic , Diterpenes , Chemistry , Pharmacology , Salicaceae , Chemistry , Classification , Flavonolignans , Chemistry , Pharmacology , Glucosides , Chemistry , Pharmacology , Hypolipidemic Agents , Pharmacology , Molecular Structure , Plant Extracts , Pharmacology , Plants, Medicinal , Chemistry , Classification
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