| Objective:The VVC,commonly and frequently occurs among women of child-bearing age,is difficult for patients to recover and affect a majority of patient's daily life.Currently clinical treatment for the VVC adopt different antifungal drugs and treatments,but the antifungal drugs and treatment is in clinic limited for its tolerance and side-effect.Traditional Chinese medieine has its unique advantages on treatment of dermal mucous fungal disease for its wide sources,low cost,less side-effect and resistance.This thesis observes Bacteriostatic Test of candida albicans and Content Determination of partial Chinese listed gynecological lotion and and hospital preparations,in order to study the relation between anticandida activity and Content Determination.And found that such lotions are low in curative effect for their low content of available component at present.In order to improve the existing defects,this thesis attempted to screen out the optimal inhibitory effects of candida albicans by evaluating the invitro anti-candidal activity of single Chinese herbs extracts.The single Chinese herbs,extracted in different means,were widely used in descriptions of both ancient and modern clinical treatment of VVC.Under the guidance of Traditional Chinese medicine formulating principles,we prepared the prescription by the way of optimizing the formulation,preparation technology and establish the quality standard.And then observe Bacteriostatic Test of candida albicans by determining Minimal inhibitory concentration(MIC)and Mininal bactericidal concentration(MBC)and to explore the antifungalm echanisn of the KBN Thixotropic gel,Which provides the foundation for successive study.Methods:1.Research on Bacteriostatic Test of candida albicans and Preparation conditions of partial listed gynecological lotion and hospital preparations.In this section,The in vitro antibacterial efficacy,the contents,compatibility and drugs of partial listed gynecological lotion and hospital preparations were annalyed for the further research.2.To screen the preliminary effects of candida albicans of single Chinese herbs extracts and prepare the KBN prescription.In this section.We used the K-B paper method to screen out the optimal invitro anti-candidal activity of single Chinese herbs extracts,which extracted in different means and were widely used in descriptions of the ancient and modern clinical treatment of VVC.Under the guidance of Chinese medicine theory,we prepared The KBN prescription by the way of optimizing the formulation.3.Preparing technology.In this section,we adopt the orthogonal design with inhibitory effects of candida albicans to optimize the extraction method.To optimize the molding formula of KBN Thixotropic gel,types and assorted dosages of drug carrier were studied.4.The quality standard.TLC was used to determine the Chinese herbs in KBN Thixotropic gel and HPLC was adopted to determine the contents of the Rhizoma Coptidis alkaloids,eugenol and Sauchinone.And we has studied the examinations of gels quality control.5.Antifungal susceptibility of KBN preparation to clinical isolations a broth microdilution tests and electron microscopy.The minimal inhibitory concentration(MIC)and the minimum bactericidal concentration(MBC)of KBN preparation Thixotropic gel was determined by broth dilution method.Ultra-structural changes of Calbicans were observed by electron microscopy before and after KBN preparation treatment.Results:1.Different kinds of gynecological lotion had inhibitory effects in different degree to candida albicans.Fuyanjie containing chemical medicines has the strongest affection.Then were the pure Chinese medicinal preparations,Huangku Xiji,Fufangkushen Xiji,Xianglian Waixiye and Xianghe Xi ji,with high content of Berberin and 4 kinds of alkaloids in coptis.The inhibitory effects of Jieeryin Xiye,Huangpujiefu Xiji,Xiaoyanzhiyang Xiji and Kubai Xiye were lower with lower content of Berberin and 4 kinds of alkaloids in coptis,either.The prescription,and the content of index component and crude drug may have influence on its antimicrobial action.2.This section screen out the optimal invitro anti-candidal activity of single Chinese herbs,such as Coptis chinensis,Syzygium aromaticum,radix sophorae flavescentis,radix stemonae,saururus chinensis,Folium Isatidis,cockscomb,etc.Then we prepared The KBN prescription by the way of optimizing the formulation for subsequent experiments.3.To establish the determining method of alkaloids from coptis root,Sauchinone and eugenol HPLC,which is used for selecting the carrier type and determining process.The optimized parameter by the orthogonal design were as follows:eight times amount of 80%ethylalcohol as percolation solvent,6 ml/(min.kg)as the rate of percolation.The optimized parameter of moulding method were as follows:4.0%Avicel CL-611 was used as carrier,which can increase drug loading especially the content of Rhizoma coptidis.4.About the quality standard,we indentified the Chinese herbs in KBN Thixotropic gel by TLC and determined the contents of the Rhizoma Coptidis alkaloids,eugenol and Sauchinone in KBN Thixotropic gel by HPLC.And we has studied s examinations of gels quality control,such weight deviation,as pH,viscosity,and microbe limitation test.5.The the minimal inhibitory concentration(MIC)and the minimum bactericidal concentration(MBC)of KBN Thixotropic gel against 3 strains of Candida Albieans in vaginal isolates show that the KBN preparation had optimal inhibiting effect of the clinical isolations.From the photograph observed by TEM,it was seen that after treatment with KBN Thixotropic gel,the morphology and ultrastructure of Candida albicans showed obvious changes:their cell wall and cytoplasmic membrane ruptured,intracellular components leak out,organelle swell,dissolve and formed large blank region.There were no changes in the morpholog after treatment with KBN Thixotropic gel.Conclusion:1.The prescription,the content of index component and crude drug in listed lotion may have influence on its antimicrobial action.2.The invitro anti-candidal activity could be enhanced by means of increasing the the concentration of Rhizoma coptidis.3.The Avicel can increase the stability and uniformity of the KBN preparation,which was unstable for the increase concentration of Rhizoma coptidis.4.The characteristic spots of Chinese herbs,identified by TLC,was clear and the reproduecibility was nice,so the TLC can be used to set up qualitystandard.The determination of contents of the Rhizoma Coptidis alkaloids,eugenol and Sauchinone in KBN Thixotropic gel by HPLC had accurate result,high sensitivity and nice reproducibility,and it can be used to set up quality standard.According to determining results of 3 batches of Pilot sample,we tentatively defined that the contents of the Rhizoma Coptidis alkaloids,eugenol and Sauchinone of KBN Thixotropic gel were not less than8.45mg,0.72 mg,0.08mg respectively in per gram.5.The data indicates that KBN Thixotropic Gel has antifungal effect on Candida albican and thus has great clinical importance.The antifungal mechanism of KBN Thixotropic Gel may be that the structure of cell wall and cell membrane of Candida are inhibited.This paper completed The KBN lotion component,preparation technique,quality standard and antifungal susceptibility of KBN preparation to clinical isolations a broth microdilution tests and electron microscopy.And we increased drug loading especially the content of Rhizoma coptidis by adopting Thixotropic Gel,which provide a foundation for further clinical observation. |
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