Application of Weiterone in the preparation of medicines for treating or preventing hand, foot and mouth disease
A technology for hand, foot and mouth disease and weiterone, which is applied in the field of medicine, can solve problems such as teratogenicity and inhibition of growth and development, and achieve the effects of relieving symptoms, suppressing the release of inflammatory factors, and prolonging survival time
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Embodiment 1
[0029] The Vero cytotoxicity of embodiment 1 Weiterone
[0030] 1. Materials and methods
[0031] 1.1 Cells and culture methods
[0032] African green monkey kidney cells (Vero cells) were used as the cell model (provided by the Institute of Microbial Epidemiology, Academy of Military Medical Sciences, the same below), and DMEM medium (purchased from Jiangsu) containing 10% fetal bovine serum (purchased from Gibco Company) was used KGI Biotechnology Co., Ltd.) at 37°C, 5% CO 2 Cultivate in an incubator, and subculture when the cells grow to 90% density, and the ratio of cell subculture is 1 / 3-1 / 4.
[0033] 1.2 Reagents
[0034] MTS Cell Proliferation Quantitative Detection Kit (purchased from Promega); Weiterone (purchased from Shanghai Yuanye Biotechnology Co., Ltd., the same below).
[0035] 1.3 Instruments
[0036] Microplate reader (purchased from Perkin Elmer Company, model: EnSpire, the same below); inverted microscope (purchased from Olympus Company, model: CKX41, ...
Embodiment 2
[0047] Embodiment 2 Weiterone protects Vero cells infected with EV71 and CoxA16 virus and inhibits viral replication
[0048] 1. Materials and methods
[0049] 1.1 Cells and culture methods
[0050] African green monkey kidney cells (Vero cells) were used as the cell model, and DMEM medium containing 10% fetal bovine serum was used at 37°C and 5% CO 2 Cultivate in an incubator, and subculture when the cells grow to 90% density, and the ratio of cell subculture is 1 / 3-1 / 4.
[0051] 1.2 Virus strains
[0052] EV71 virus BJ09 / 07 strain, GenBank accession number JQ319054.1; CoxA16 virus TS10 / 08 strain, GenBank accession number JX068829, all provided by the Institute of Microbial Epidemiology, Academy of Military Medical Sciences, the same below. Determine the half cytopathic dose (TCID) of EV71 virus before use 50 ) is 10 8 / mL, CoxA16 TCID 50 for 10 8.5 / mL.
[0053] 1.3 Reagents
[0054] MTS Cell Proliferation Quantitative Detection Kit; Weiterone; Total RNA Extraction ...
Embodiment 3
[0073] Embodiment 3 The broad-spectrum research of weiterone anti-hand-foot-mouth virus
[0074] 1. Materials and methods
[0075] 1.1 Cells and culture methods
[0076] African green monkey kidney cells (Vero cells) were used as the cell model, and the cells with the number of passages between 130 and 145 were used in the experiment. Cells were cultured in DMEM medium containing 10% fetal bovine serum at 37°C, 5% CO 2 Cultivate in an incubator, and subculture when the cells grow to 90% density, and the subculture ratio is 1 / 3-1 / 4.
[0077] 1.2 Virus strains
[0078] Coxsackieviruses A groups 4, 5, 7, 9 and 10 (CoxA4, CoxA5, CoxA7, CoxA9 and CoxA10), Coxsackieviruses B groups 2 and 5 (CoxB2 and CoxB5) and echoviruses (ECHO) . Measure the TCID of each virus before use 50 10 respectively 8 、10 7.5 、10 7 、10 6 、10 8.5 、10 7.5 、10 7 and 10 7.5 / mL, the above viruses were provided by the Wuhan Institute of Virology, Chinese Academy of Sciences.
[0079] 1.3 Reagents ...
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