Objective:To study the sensitivity of pathogenic fungi from otorhinolaryngology to 5 antifungal agents of Ketoconazole(KE), Itraconazole(IT),Fluorocytosine(FC),Amphtericin B (AP) and Fluconazole(FL) by Etest method.Method:Etest for determine MIC of 14 pathogenic fungi was performed according to the manufactuer′s instructions.The Etest strip containing the 5 antifungal drugs were putted on medium with Candida albicans,A.flarus and A.fumigatus etc,respectively,to determine MIC of antifungal drugs.Result:Candida albicans were sensitized to all the above 5 antifungal drugs, KE and IT were most sensitized.Seven fungi such as A. flavus, A fumigatus, A.oryzae etc were all sensitized to KE and IT.The MIC range of KE and IT against 15 strains of pathogenic fungi were≤0.008～2 mg/L and ≤ 0.006～4 mg/L respectively.In 15 strains,11 strains showed drug-resistant to FL,8 strains to AP and FC,4 strains to KE and 1 strains to IT respectively and MIC were all >32 mg/L.Conclusion:Etest MIC were in good agreement with macrobroth MIC.The use of Etest strips for antifungial susceptibility test is a new and promising method with advantage of easy to perform, exact results and satisfactory reproducibility. Etest is valuable on clinical practice.

BACKGROUND:Study shows that margarita liquid has effect on promoting histiocyte regeneration and removing scars.OBJECTIVE:To observe the effects of margarita liquid on the proliferation of fibroblasts in human skin scar tissues.DESIGN,TIME AND SETTING:A grouping contrast observational experiment was performed in the Experiment Centre of Guangxi Medical University from September to December in 2008.MATERIALS:Scar tissue samples were obtained from patients in the Department of Plastic Surgery,Guangxi Medical University.Margarita liquid was the digest of margarita purchased from Beihai Gofar Marine Biological Industry Co.,Ltd and rich in multi-amino acids,polypeptides,vitamin,mineral matters and natural enzymes.METHODS:Human skin scar cell line was established by removing non-fibroblasts through repeated primary culture and serial subcultivation of scar fibroblasts with reference to Veelken method. The 3-5 generation human skin scar fibroblasts on exponential phase of growth were made into single cell suspension by trypsin digestion which was then inoculated on plastic 96-well cell culture plate,with the density of 0.5×10~4/well as well as 100 μL cell suspension and 100 μL DMEM medium in each well. After culture for 24 hours,primary medium was discarded. The grouping of the experiment:200 μL mediums with 125.00,62.50,31.30,15.60,7.80,3.90,1.95,0.98 mg/L margarita liquid were used respectively in margarita liquid group;200 μL pure medium was added into each well in control group.MAIN OUTCOME MEASURES:MTT and TUNEL assay were used to examine the proliferation and the apoptosis of fibroblastsrespectively.RESULTS:The 50% inhibiting concentration (IC_(50)) of margarita liquid on fibroblasts was 15 mg/L. Margarita liquid at any other concentration but 0.98 mg/L was effective in inhibiting fibroblast growth in a dose-dependent way,i.e. the higher margarita liquidconcentration,the higher inhibition ratio on fibroblast growth. Fibroblasts cultured with 15 mg/L (IC_(50)50) margarita liquid had got reduced volume,lessened cytoplasm,decreased density,increased apoptosis rate and buffy colour. Fibroblasts in control group were large,rich in cytoplasm and compact. Apoptotic index was higher in the margarita liquid group than in the blank control group CONCLUSION:Margarita liquid could inhibit the proliferation of skin scar fibroblasts cultured in vitro and induce the apoptosis of them.

ObjectiveTo test the susceptibility of Penicilliosis marneffei (PM) isolates from Guangxi bamboo rats and patients to voriconazole and several commonly used antifungal agents.MethodsAccording to the Clinical and Laboratory Standards Institute(CLSI) M27-A2 and M38-A document,a microdilution method was used to determine the minimum inhibitory concentration(MIC) of voriconazole,itraconazole,terbinafine,amphotericin B,and fluconazole against mycelial phase (25 ℃) and yeast phase (37 ℃) of 14 PM isolates from Guangxi Bamboo rats and 25 PM isolates from patients.The difference in MIC of the antifungals was assessed by two-sample t test between Bamboo rat PM isolates and clinical PM isolates,and by paired t test between the mycelial and yeast phase of PM isolates.Results The MIC ranges of voriconazole,itraconazole,terbinafine,amphoteriein B and fluconazole were 0.0313-0.1250,0.1250-1.0000,0.0313-0.5000,0.2500-4.0000,2.0000-8.0000 mg/L,respectively for mycelial phase of Bamboo rat PM isolates,0.0078-0.2500,0.0313-0.5000,0.0313-1.0000,0.2500-2.0000,1.0000-8.0000 mg/L,respectively for yeast phase of Bamboo rat PM isolates,0.0313-0.2500,0.0625-1.0000,0.0313-1.0000,0.2500-4.0000,2.0000-32.0000 mg/L,respectively for mycelial phase of clinical PM isolates,0.0039-0.2500,0.0313-0.5000,0.0313-2.0000,0.1250-2.0000,2.0000-16.0000 mg/L,respectively for yeast phase of clinical PM isolates.None of the PM isolates was resistant to any of the antifungals.The MIC of voriconazole was found to be the lowest for PM isolates from both Bamboo rats and patients at the same temperature (37 ℃ or 25 ℃),followed by itraconazole,terbinafine,amphotericin B and fluconazole.Statistical difference was found in the MIC values of itraconazole,terbinafine,amphotericin B between the yeast and mycelial phase of the same PM isolate,but not found in antifungal MIC values between Bamboo rat isolates and clinical isolates at the same phase.ConclusionsOf the tested drugs,voriconazole shows the strongest antifungal potency. The PM isolates from Guangxi Bamboo rats are similar to clinical PM isolates in the sensitivity to voriconazole,itraconazole,terbinafine,amphotericin B and fluconazole.The phase of PM isolates may affect their susceptibility to itraconazole,amphotericin B and terbinafine.

Objective To evaluate the preventive and therapeutic effects of artemisinin and artesunate on hypertrophic scar in rabbit ears.Methods Full-thickness wounds to cartilage were created in New Zealand white rabbit cars to establish animal models of hypertrophic scar.Cream was prepared with artemisinin or artesunate.A total of 96 hypertrophic scars were divided into 4 groups to receive the treatment with artemisinin cream.artesunate cream.cream vehicle(vehicle control)or no treatment(blank control)28 days after wounds were created.After 28-day treatment,animals were scarified,scars were incised and examined with HE-staining and VG-staining.Hypertrophy index.numerical density of fibroblasts and area density of collagen fibers were calculated.Results Compared with vehicle and blank controls,the scars were softer and flatter,the volume of fibroblasts decreased,and collagen fibers appeared to be more regulated and sparse in artemisiIlin or artesunate cream-treated groups.The Hypertrophy index.numerical density of fibroblasts.area density of collagen fibers were(1.452±0.27),(3638.245±463.0)cells/mm2,(32.29±6.9)%in artemisinin cream-treated group,respectively,(1.445±0.24),(3585.016±638.9)cells/mm2,(34.74±8.27)%in artesunate cream-treated group,respectively.All the three parameters were significantly reduced in artemisinin and artesunate groups than in blank and vehicle control groups(all P＜0.0 1).but no significant difference was found between artemisinin and artesunate groups (P＞0.05).Conclusion Artemisinin and artestmate cream has a reliable efficacy in the prevention and treatment of hypertrophic scar in animal models.