| Rabbit Dermato-mycosis that caused by a variety of skin filamentous bacteria is one beast contract Dermato-mycosis that have strong infectivity.And then its hazard is very serious.The main common pathogen are Micros porum canis,Trichoph yton mentagroph ytes and Micros porum gypseum. In recent years,the slather of antifungal drug chemistry with a strong side effects that have caused the rapid development of fungal resistance.But a lot of natural medicine with smaller side effects have antifungal activity. In order to effectively making use of the rich resources of traditional Chinese medicine(TCM),this study selected a few of TCM with stronger antibacterial activity in vitro from 20 TCM. And study their joint bacteriostatic activity,then make them with no antagonism effect preparations.Last observe its clinical efficacy and toxicity.The related specific research results are as follows:1.The study of antibacterial activity in vitro of 20 TCM:First extract their antibacterial active. site through ethanol reflux,water decoction and ultrasonic methods.Next determine their antibacterial activity in vitro against Micros gypsum gypsum such as four strains adopting Broth Microdilution Susceptibility Method and study the joint bacteriostatic activity of a few of TCM with stronger antibacterial activity in vitro.The results:the effective parts of Belamcanda chinensis,Dictamnus dasycarpus Turcz, Melaphis chinensis and Sophora flavescens Ait have the strongest antimicrobial activities and the range of MIC is 1.563～12.5 mg/ml; the effective parts of Curcuma longa L., Eucommia ulmoides Oliv., Paeonia lactiflora Pall., Gardenia jasminoides Ellis, Forsythia suspensa (Thunb.)Vahl and Bupleurum chinense DC. have stronger antimicrobial activities and the range of MIC is 6.25～50 mg/ml.We selected Belamcanda chinensis, Melaphis chinensis, Curcuma longa L. and Gardenia jasminoides Ellis that have good antimicrobial activities against fungi and reported less. And then study their joint bacteriostatic activity.The fractional inhibitory concentration index of Belamcanda chinensis and Curcuma longa L., Gardenia jasminoides Ellis are less than or equal to 2.The fractional inhibitory concentration index of Melaphis chinensis and Belamcanda chinensis, Curcuma longa L., Gardenia jasminoides Ellis are greater than 2.The results show that Melaphis chinensis and Belamcanda chinensis, Curcuma longa L., Gardenia jasminoides Ellis have antagonistic action and Belamcanda chinensis and Curcuma longa L., Gardenia jasminoides Ellis have independent, additive or synergy action.2.The technical identification of traditional Chinese medicine and its extraction technology:According to the specified method in "Chinese pharmacopoeia" 2010 version of the book,by thin layer chromatography,identify the API’s for the screening out no antagonism effect of Curcuma longa L. Belamcanda chinensis(L)DC. and Gardenia jasminoides.With extraction rate and MIC as the index, the following five factors were considered:extraction method,concentration and dosage of extraction solvent,extracting times and extracting time, the extracting process of active substance in three traditional Chinese medicine were optmized by the orthogonal.The results:In the three chromatogram of the test,they showed the same spot at the corresponding position of contrast medicinal chromatographic and reference substance chromatography,The optimization conditions of the extracting process were that:the coarse powder of Curcuma longa L.(20sieve),80% ethanol(6 times)as solvent, heating reflux extraction 3 times, each time 2h;the coarse powder of Belamcanda chinensis(L)DC.(20sieve),70% ethanol(10 times)as solvent, heating reflux extraction 3 times, each time 2h;the coarse powder of BGardenia jasminoides Ellis(20sieve),80% methanol solution(6 times)as solvent,soaking half an hour and then ultrasonic extraction 2h.The results show that Curcuma longa L. Belamcanda chinensis(L)DC. and Gardenia jasminoides,such as the three TCM,all meet the standard herbs in pharmacopoeia.Under the optimal extraction process,the MIC of three kinds of extracts that be of three strains of fungi are respectively 31.25mg/ml、25mg/ml、18.75mg/ml, and that bacteriostatic effect is obvious and the extraction process is stable,feasible.So it can be widely used for the quality control of active substance in four traditional Chinese medicine.3.The study of the optimum preparation process of the prescription of Curcuma longa L. Belamcanda chinensis(L)DC. and Gardenia jasminoides Ellis in plastics:Through uniform experimental design, valuing the MIC of joint antibacterial activity in vitro as reference indexes, we selected their best matching is 4:3:2.On the basis of the best matching,set up the low,medium and high three doses main medicine in formula temperture.And then selected conditions of each temperture through orthogonal experimental design and developed three doses formula temperture.To measure its content of curcumin by high performance liquid chromatography.The results showed that the best matching of Belamcanda chinensis and Curcuma longa L., Gardenia jasminoides Ellis is 3:4:2.The recipe of three doses formula temperture as follows:The amount of film forming solution is PVA1788 8%,glycerin 2%;three main medicine mixed according to the best matching the adding proportion are respectively 2.25%、6.75%、20.25%;the adding proportion of solubilizer twain-80 are respectively 4%、 5%、5%;the adding proportion of transdermal promter are 5%1,2-Propanediol and 8% Bomeol.According to the best preparation technology,the content of curcumin in the low、medium and high doses formula of film coating are respectively 1.62μg/ml、3.74μg/ml、7.82μg/ml.4.The research of the prescription of Curcuma longa L. Belamcanda chinensis(L)DC. and Gardenia jasminoides Ellis in plastics that resist rabbit ringworm fungi infection:The rabbits was infected with standard stain of Microsporum canis. Then, the scurf and coat from the infected rabbits were collected for selecting the lowest concentration with which the fungi could result in infection. The infection model was established with the lowest concentration of fungies. After that, the liniment prescriptions consisted of Curcuma longa L, Belamcanda chinensis(L)DC. and Gardenia jasminoides Ellis at three different concentrations (2.25%、6.75%、20.25%)were applied for the treatment. The infection model was established with the Microsporum canis 1×108 CFU/ml. In the activity assays, the results showed that the prescriptions at the concentration of 6.75%、20.25% exerted a positive effect on diminishing the dermatophytosis caused by Microsporum canis. Moreover, the infection was totally cured with the treatment of the prescriptions at the concentration of 20.25%. The results of this study indicated that the prescription of Curcuma longa L, Belamcanda chinensis(L)DC. and Gardenia jasminoides Ellis have additively curative effect on fungal skin infection on rabbits.5. Safty evaluation of the prescription of Curcuma longa L. Belamcanda chinensis(L)DC. and Gardenia jasminoides Ellis in plastics:Considering the possibility such as licking,acute oral toxicity test of the prescription of Curcuma longa L. Belamcanda chinensis(L)DC. and Gardenia jasminoides Ellis in plastics was performed.Modified Kaeber’s method was used for median lethal dose(LD50); meanwhile,based on the property of an external medicine.acute skin toxicity test,skin allergic.skin irritation experiment,eye irritation and mucosal stimulation experiment of this plastics were conducted to study the toxicology.The result.In the acute oral and skin toxicity tests of formula temperture,when the dose is 5000mg/kg,the mice and rabbits are still no deaths,and so it is non-toxic drug.The formula temperture caused no obvious erythema and edema stimulus response to the intact skin and the damaged skin,and caused no skin anaphylaxis.The stimuli mucosa reaction of rabbit eyes and degree of oral to formula temperture is without excitant. The result showed that the prescription of Curcuma longa L. Belamcanda chinensis(L)DC. and Gardenia jasminoides Ellis in plastics is safety drug and can be used for a long time in the treament of rabbit skin tinea disease. |
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