Preparation And Pharmacodynamic Study Of The "YAJIANGXIN" Injection

Riemerella anatipestifer disease,which is caused by Riemerella Anatipestifer RA),is an acute or chronic hematosepsis among ducks.It is arch-CD in duck industry with high attack rate and death rate.Over the years,the routine treatments for riemerella anatipestifer disease are antibiotics and sulfonamide-related medicine.However,studies have shown that the heavy use of antibiotics has caused severe drug fast phenomenon donec,which have negative impact on their effectiveness.In the research,we were able to categorize RA disease as lung anthracia from the aspect of Chinese traditional veterinary theory by analyzing the data from the area of clinical symptoms,incidence,and pathological changes.Furthermore,based on the results of anti-bacteria experiments,a novel injection medicine named "Ya Jiang Xin" was developed from Chinese traditional medicine.Detailed study on its quality criteria and partial study of its pharmacodynamics was carried out using bacteriostasis experiment.The results are summarized below:1.The best recipe and composition of this novel injection medicine were determined based on pathogenesis obtained through differentiation of symptoms and signs,using the results of bacteriostasis experiment as the additional guidance.Among three anagraphes studied,the anagraph 3,with the composition of baical skullcap root, forsythia suspense and isatidis=1:2:1,ft.injection containing crude drug 0.5g/mL,was the best.The data of the bacteriostasis experiment of the anagraph 3 under 10-fold dilution was 20.5±1.2mm,equivalent to that of ciprofloxacin(2μg/mL).MIC from the anagraph 3 was 7.8mg/mL.Based on the results above,it was concluded that anagraph 3 was the final optimal anagraph.2.The amount of baicalin in the novel "Ya Jiang Xin" injection was detected by high performance liquid chromatography(HPLC):Chromatographic condition:Chromatography column:Ultimate XB-C18,5μm,4.6×150mm;Detector: ultraviolet detector;Mobile phases:colonial spirit-glacial acetic acid- aquaefer(50+1+50,V/V);Flow rate:1mL/min;Column temperature:35℃;UV Det.W/L:274nm;Sample size:10μL.Procedure:1mL sample solution was prepared in the measuring flask(50mL).It was diluted to the desired concentration using HPLC mobile phases,mixed thoroughly,and then filtered through membrane(0.45μm) to get the clear sample solution.10μL sample was injected on to the HPLC,the data was analyzed and the concentration was calculated by measuring each peak area against the standard peak area-concentration chart.This HPLC method is very consistent and reliable with good reproducibility and high recovery rate.It could be served as one of the standard detecting methods for this novel injection medicine in future manufacturing.3.Detecting the organic acid cont.in "Ya Jiang Xin" Injection by electric titration.Procedure:To the sample solution(2mL) was added 50%alcohol solution(10mL) and 0.101moL/L NaOH solution(10mL).The resulting solution was then thoroughly mixed and put on magnetic stirrer.Combination electrode was inserted and it was connected with a pH meter.By following the standard titration procedure,the sample solution was titrated using 0.111mol/L HCl solution.Changes of pH were recorded, which were used to draw the first order derivate curve[(△pH//△V)-V].The apogee of curve was the VOL of titration end point.The content of the organic acid was then worked out through the curve. This method can be used as one of future detecting method for organic acid based on its stabilization,reliability,high reproducibility,and highly recovery rate.4.Monitoring clinical symptoms and the changes of TNF-a cont.and IL-1 cont.in the blood plasma of poultry ducks,which were infected with RA under experimental conditions.By comparing our experimental results with literature reports in the area of biological function of TNF-a and IL-1,we confirmed the quick release of large amount of TNF-a into blood would produce gen-inflammatory reaction syndrome,induce the formation of other inflammatory factors and tissue damage in many organs;On the other hand,the quick release of large amount of IL-1 could produce febrility and lethargy by acting on endocrinium,it also could decrease the animal's cibation by acting on hypothalamus appestat.As the result,various clinical symptoms and pathological changes were demonstrated on infective animals because of the effects of the excess quick release of TNF-a and IL-1.5.By monitoring the changes of TNF-a and IL-1 cont.in blood plasma of RA-infected poultry ducks under the treatment of new "Ya Jiang Xin" injection and under the treatment of traditional western,we confirmed that "Ya Jiang Xin" Injection could cure the RA disease as effectively as other traditional medicines.The using of this injection in the treatment not only killed the bacterium,but also possessed other advantages, such as anti-toxicity ability against endotoxin from RA,effectively prohibiting the release of inflammatory factors,protecting the cell endothelium from traumatize,and raising the immune function of organism.Thus,the above mentioned pathogenic actions lay down the fundamental functional mechanism of this novel injection to cure RA disease.