Compound traditional Chinese medicine for effectively preventing and controlling H1N1 and H3N2 swine influenza and porcine reproductive and respiratory syndrome, and preparation method thereof
A technology for porcine blue-ear disease and swine flu, which is applied in medical preparations, pharmaceutical formulations, antiviral agents and other directions containing active ingredients, can solve the problems that have not been researched and cannot fully evaluate the effect of porcine reproductive and respiratory syndrome virus.
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Embodiment 1
[0045] Example 1 In vitro anti-porcine reproductive and respiratory syndrome virus effect of Chinese medicine compound preparation
[0046] 1. Determination of the median lethal dose (TCID50) of PRRSV on MARC-145 cells
[0047] The virus to be tested was serially diluted 10 times in 10 concentration gradients, and the virus solution was added to a 96-well plate full of Marc-145 cells at a dose of 100 μl / well, and each gradient was repeated 8 times, and the last 2 columns were For the cell control group, the virus liquid was discarded after adsorption at 37°C for 1.5 h, and 100 μl of maintenance liquid was added, and cultured in a CO2 cell incubator.
[0048]The cell lesion was observed every day, and the OD value of each well was measured by the MTT method after the lesion reached 75%. Growth inhibition rate of virus on cells = (OD value of cell control group - OD value of virus control group) / OD value of cell control group. The median toxic concentration (TCID50) of the v...
Embodiment 2
[0068] Example 2 Prevention and treatment effect of traditional Chinese medicine compound on mice infected with H1N1 and H3N2 swine influenza virus
[0069] 1. H1N1 and H3N2 swine influenza virus half infectious dose to chicken embryo (EID 50 ) Determination
[0070] The virus liquid was sequentially diluted 10 times, and inoculated in the allantoic cavity of 10-day-old chicken embryos, each dilution was inoculated with 0.2ml / piece, and 4 replicates were made, and normal chicken embryos were set as a control at the same time. Place them in an incubator at 37°C for 72 hours, discard the dead embryos within 24 hours, harvest the allantoic fluid of the dead embryos after 24 hours and live embryos after 72 hours, and measure the titer of HA. Calculate EID according to the Reed-Muench method 50 .
[0071] . Effects of traditional Chinese medicine compound on pneumonia in mice infected with H1N1 and H3N2 swine influenza virus
[0072] (1) Grouping of experimental animals
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