Table of Content

28 February 2011, Volume 6 Issue 1

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Combination of itraconazole and terbinafine in treating sporotrichosis and their in vitro antifungal activities against Sporothrix schenckii

WANG Ai-ping, GAO Na, LIU Wei, WAN Zhe, CHEN Wei, TU Ping, LI Ruo-yu

2011, 6(1):  5-9. 

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Objective To evaluate the efficacy of the combination of itraconazole and terbinafine in treating sporotrichosis and against Sporothrix schenckii growing in both mycelial and yeast phases.Methods The combination of itraconazole(200 mg/d) and terbinafine(250 mg/d) was applied in the treatment of sporotrichosis.In vitro effects against Sporothrix schenckii in two growth phases were evaluated by checker-board method,and fractional inhibitory concentration(FIC) index was caculated to determine whether the interaction was synergistic,indifference,or antagonistic.Results Three patients failed in itraconazole therapy were cured with the combination of itraconazole and terbinafine.Synergistic interactions of itraconazole and terbinafine against Sporothrix schenckii in mycelial form and in yeast form,were observed to be 75% and 70% respectively(P=0.50).Conclusions The combination of itraconazole and terbinafine is efficient and safe in treating sporotrichosis with perfect synergy.

Genotype analysis and antifungal susceptibility of Candida albicans in Xinjiang area

Paride Abliz, Halmurat Ghupur, Takashi Yaguchi, Kayoko Takizawa, LI Ruo-yu

2011, 6(1):  10-14. 

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Objective To analyze the genotypes of 50 isolates of Candida albicans from Han and Uyghur patients in Xinjiang area and to understand the in vitro antifungal susceptibilities of these isolates against amphotericin B,flucytosine,micafungin,itraconazole,fluconazole and miconazole.Methods The genotypes were identifed by the size of PCR amplicons amplifined from the group I intron contained region of 25S rDNA of C.albicans:genotype A(450 bp),genotype B(840 bp) and genotype C(450 bp and 840 bp).In vitro antifungal susceptibilities of 50 strains to 6 kind of antifungal agents were determined by CLSI M27-A microdilution method.Results Fifty strains were classified into three genotypes,including 30 strains of type A and 10 strains of type B and C,all these strains showed lower MIC to amphotericin B,flucytosine,micafungi,and miconazole(MIC range:0.25-0.5 μg/mL,0.125-0.25 μg/mL,≤0.03 μg/mL,and 0.25-8 μg/mL,respectively),and higher MIC to itraconazole and fluconazole(MIC range:0.25-8 μg/mL and 0.5-64 μg/mL,respectively).The MIC of the genotype B and C strains to flucytosine was 0.125 μg/mL.The resistant ratio of itraconazole and fluconazole were 84% and 70%,respectively.No statistical significance was found in the genotypes of the strains from different races(P>0.05) or different genotypic strains and antigungal susceptibilities(P>0.05).Conclusions C.albicans in Xinjiang area included 3 genotypes(A,B and C).There was no correlation between the genotype and antifungal susceptibilities of C.albicans strains from Han and Uyghur patients.Nither did genotype and race.All strains were resistant to itraconazole and flucytosine,but genotype B and C showed high susceptibilities to flucytosine.

Antifungal susceptibility of the A.fumigatus transformants containing extra copies of A.flavus cyp51 gene homologues to the common antifungal drugs

LIU Wei-xia, SUN Yi, WAN Zhe, LI Ruo-yu, LIU Wei

2011, 6(1):  15-21. 

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Objective To investigate the effect of Aspergillus flavus cyp51 genes on antifungal susceptibility by cloning and constucting the extra copies of Aspergillus flavus cyp51 genes.Methods A.flavus cyp51 gene homologues were identified by tblastn searches in A.flavus genome database.PCR fragments composed of the 5′flanking sequence(approximately 1 000 bp) of cyp51,cyp51ORF,and its 3′flanking sequence(approximately 1 000 bp),were subcloned into shuttle plasmid pRG3-AMA1-NotI to produce recombinant plasmids.These plasmids and empty plasmid pRG3-AMA1-NotI were transformed into A.fumigatus strain AF293.1(pyrG-) respectively to produce transformants.The Clinical Laboratory Standard Institute broth microdilution method M38-A2 and E-test method were used to assay the minimal inhibitory concentrations(MICs) of itraconazole(ITC),voriconazole(VRC),amphotericin B(AMB),and posaconazole(POS),or minimal effect concentration(MEC) of caspofungin(CAS),against these transformants.Results A.flavus genome contains three cyp51 gene homologues,cyp51A,cyp51B and cyp51C,of which the ORF size are 1 400-2 000 bp.When these genes were subcloned into shuttle plasmid pRG3-AMA1-NotI,we get plasmids pRG3-AMA1-CYP51A,pRG3-AMA1-CYP51B and pRG3-AMA1-CYP51C.These plasmids and empty plasmid were transformed into A.fumigatus strain AF293.1(pyrG-) to produce transformants rCYP51A,rCYP51B,rCYP51C and rpRG.The antifungal susceptibility of these A.fumigatus transformants to the antifungal drugs by broth microdilution assaying and E-test method showed that,rCYP51A and rCYP51B were cross-resistant to VRC and ITC,susceptible to both AMB and CAS;rCYP51C and rpRG were intermediate to ITC and VRC,susceptible to both AMB and CAS.Conclusion In A.fumigatus,extra copies of A.flavus′cyp51A gene or cyp51B gene have effect on antifungal susceptibility to azoles,have no effect on AMB and CAS.Extra copy of cyp51C has no obvious effect on all the tested drugs.

Effect of temperature on the growth and spore production of Fusarium solani

DONG Xian-hui, QIAN Tao, GAO Wei-juan, HE Xiao-ping

2011, 6(1):  22-25. 

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Objective To investigate the influence of temperature on the growth,sporulation quantity and pathogenicity of Fusarium solani.To find out the best way for promoting conidial production of Fusarium solani.Methods Fusarium solani from 5 different groups were incubated in dark for 7 days at 20℃,25℃,30℃,35℃,and 37℃,respectively.Group F was incubated in dark for 3 days at 35℃,and at 25℃ for the next 4 days.The diameters of the colonies were measured by vernier caliper on 24 h,3 d,5 d,and 7 d.Conidial production was determined by hematimeter on 3 d,5 d,and 7 d.Results Between The growing speed of Fusarium solani colony increased during 20℃ to 30℃,while decreased during 30℃ to 37℃ with the temperature going up.Conclusions Fusarium solani began to grow and produced conidiospore at 20 to 35℃.The pathogenicity of the Fusarium solani was affected by temperature obviously,with 30℃ as the suitable temperature.Incubation at 35℃ for 3 days and at 25℃ for consecutive 4 days was good for sporulation.

Antifungal sensitivity of two Chinese traditional drugs against Candida in vitro

JIANG Miao, HUANG Xin, SHEN Liang-liang, ZHOU Fei, TU Wei-yan, SHI Wei-min

2011, 6(1):  26-30. 

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Objective To compare the susceptibilities of Candida albicans cultured with ginseng stems and leaves saponins,Panax Pseudoginseng,fluconazole and itraconazole.Methods Susceptibilities of 60 candida isolates cultured with fluconazole,itraconazole,ginseng stems and leaves saponins and Panax Pseudoginseng was determined by M27-A Microdilution Method based on CLSI.Results The MIC₅₀ and MIC₉₀ of Candida albicans to fluconazole,itraconazole,ginseng stems and leaves saponins and Panax Pseudoginseng were 0.25 mg/L,0.25 mg/L,7.812 mg/L,15.625 mg/L(MIC₅₀),and 64 mg/L,16 mg/L,250 mg/L,250 mg/L(MIC₉₀),respectively.Ten of the total 60 isolates showed cross-resistances to itraconazole,ginseng stems and leaves saponins and Panax Pseudoginseng.Conclusions Ginseng stems and leaves saponins and Panax Pseudoginseng have similar antifungal effects on Candida albicans.CLSI-M27-A could be used to evaluate the anti-candida activity to the components of medicinal plant as a standard method.

Effect of Cryptococcus neoformans on the viability of keratinocytes

ZHU Yuan-jie, GU Ju-lin, CHEN Jiang-han, ZHAO Jin, QIU Yun, WEN Hai

2011, 6(1):  31-34. 

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Objective To investigate the effect of Cryptococcus neoformans on the viability of keratinocytes.Methods Strains of Cryptococcus neoformans B3501 and its acapsule mutant cap64 were cocultured with keratinocytes and the viability of the keratinocytes was measured by flow cytometry after 0.5 h,1 h and 2 h.Results were compared with those of negative control,hear-killed fungi,and the fungi indirectly contacted with keratinocytes.Results Viability of keratinocytes decreased significantly cocultured with Cryptococcus neoformans by time prolonging,while more significantly after 1 h and 2 h’s coculture with B3501.There was no difference in the viability of keratinocytes between coculture with killed B3501 and indirect coculture with B3501.Conclusions Capsule of Cryptococcus neoformans plays an important role in viability of keratinocytes.Indirect contact with Cryptococcus neoformans is necessary for viability decreasing.

Retrospective analysis of primary cutaneous cryptococcosis from 2000 to 2008

YANG Ming-Hui, , WEN Hai, ZHU Yuan-jie

2011, 6(1):  35-39. 

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Objective To study the epidemiology and clinical features of primary cutaneous cryptococcosis from 2000 to 2008.Methods Cases of primary cutaneous cryptococcosis from 2000 to 2008 were retrospectively analysed and the differences with those between 1985 and 2000 were compared.Results PCC occurred more commonly in the older and the male in the recent 8 years.There was no difference in the onset between immunocompetent and immunocompromised hosts,while HIV positive patients had the lower probability to get PCC,xternal injury might be the important causative factors,erythra still happened majorly in extremities or other exposure sites,the erythra had changed from whitlow or cellulitis to ulcer,nodus,erythema,and so on,the oral antifungal drugs combined with surgery were most acceptable treatment and fluconazole was still the first choice for PCC,and the prognosis was good.Conclusions In recent years,PCC obtained some new characteristics.We considered that the immune state affected less or had no effect on incidence of PCC,while majorly affected the final outcome of Cryptococcus infection.

A case of fungal peritonitis caused by Humicola fuscoatra

WANG Peng, XIE Xiu-li, WANG He, DOU Hong-tao, SUN Hong-li, WANG Hui, XU Ying-Chun

2011, 6(1):  40-42. 

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A case of fungal peritonitis in CAPD patient caused by Humicola fuscoatra was reported.Characteristic of the pathogen,which widely spreads in the nature but rare in human infection was described and molecule examination was carefully performed.MICs of itraconazole,voriconazole and amphotericin B were 0.008 μg/mL,0.016 μg/mL and 1.5 μg/mL,respectively.The patient showed improved condition and was discharged from the hospital by oral itroconazole(0.1 g,q12 h) and hemodialysis.

Actinomycetic mycetoma caused by Nocardia ninae sp.nov

GUO Yun, FAN Yin-jun, ZHOU Xiao-hong, LI Wen-hua

2011, 6(1):  43-45. 

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To report a case of actinomycetic mycetoma caused by Nocardia ninae.A 37-year-old female was admitted for erythema,papulopustule,nodus,sinuses and keloidal scars in the upper thoracic wall for up to 20 years.The first lesion appeared in the middle of right clavicle,and spreaded graduately to the upper thoracic wall and lower neck,with adventitious mild pain.Erythema,papulopustules,nodus,sinuses and keloidal scars were found in the thoracic wall and neck.Sulphur grains were determined by biopsy.Staphylococcus epidermidis and Nocardia sp.,with 98.1% simlarity to Nocardia ninae OFN 02.72T was identified by sequence analysis.The final diagnosis was actinomycetic mycetoma cauesed by Nocardia ninae.

Tinea corporis caused by Microsporum gypseum misdiagnosed as Ganuloma annulare:one case report

GUO Yan-yang, QI Xian-long

2011, 6(1):  46-47. 

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To report one case of tinea corporis caused by Microsporum gypseum which was misdiagnosed as Ganuloma annulare.The patient was a 60-year-old female with erythema on the left wirst for 2 months.She was diagnosed as Ganuloma annulare but showed no response to the treatment.Tinea corporis caused by Microsporum gypseum was finally diagnosed by fungal examination.The patient was cured by one-week intraconazole(200 mg/d×7 d) combined with one-week sertaconazole nitrate ointment treatment.