Chinese Journal of Mycology

Compare extracellular proteinase activity of three phynotypes of Penicillium marneffei

WANG Peng, RAN Yu-ping, YIN Bin, ZHUANG Kai-wen, LIN Xin-yu, DAI Ya-ling

2014, 9(4): 193-198.

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Objective To find the effective way to induce Penicillium marneffei (PM) to secrete extracellular proteinase and compare the extracellular proteolytic activity of three phynotypes of PM.Methods PM phynotypes (M/M and M/Y) and their parent strain B-6323 (P) were respectively inoculated with same amount into separate yeast carbon base (YCB) agar plates containing bovine hemoglobin (BHG) as the proteolytic substrate which had been adjusted to pH 4.0, pH 5.6, pH 7.2, and incubated at 25℃ and 37℃ for 6 days. The plates were stained with Coomassie blue G-250 to be observed clearly. Clear zones surrounding each clone result from broken down hemoglobin were measured to ascertain whether YCB-BHG agar can induce PM producing proteinase successfully, and to determine the best suitable condition for PM proteinase induced and compared the proteinase activity produced by the three isolates.Results YCB-BHG agar can successfully induce PM producing proteinase. Any three phynotypes of PM inoculated into medium at any three value of pH, the diameter of clear zones surrounded each clone increased greatly at 37℃ compared with at 25℃. It had statistically significant difference by t-test; The variance analysis showed that the diameters of clear zones surrounding each clone inoculated at pH4.0, 5.6, 7.2 medium at 37℃ had statistically significant difference (F-Values 97.198,P=0.0001). Student-Newman-Keuls (SNK) method was used to compare one to another. The diameters of clear zones surrounded the clone inoculated at pH4.0, pH5.6 medium and the clone inoculated at pH7.2 medium had statistically significant difference; No matter the culture temperature was 37℃ or 25℃, in any pH medium group, the M/M strain had greatest mean diameter of clear zone, then P strain, M/Y strain was lowest.Conclusions Extracellular proteinase of Penicillium marneffei growed at 37℃ in acidic medium (pH4.0 or pH5.6) showed greater proteolytic activity. The proteolytic activity produced by different morphotypes of Penicillium marneffei was different. The extracellular proteolytic activity of M/M strain and P strain was greater than M/Y strain after growth on the same condition.

Identification on two yeasts CICC32991 and CICC32993 isolated from Paracel Islands wild Noni (Morinda citrifolia L.) Seeds

BAI Fei-rong, LIU Yang, YAO Su, LI Hui, TAN Wang-qiao, CHENG Chi

2014, 9(4): 199-202.

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Objective To distinguish the two yeasts (CICC32991,CICC32993) isolated from Paracel Islands wild Noni (Morinda citrifolia L.) seeds by the polyphasic taxonomy methods.Methods This study included morphological, physiological and biochemical characteristics of the test, as well as ribosomal DNA sequence phylogenetic analysis technology.Results Morphological and physiological test results showed the strain CICC32991 as Eremothecium sp. and CICC32993 as Kodamaea ohmeri.Phylogenetic analysis results showed CICC32991 as Eremothecium coryli and CICC32993 as Kodamaea ohmeri.Conclusion The results certified the strain CICC32991 as Eremothecium coryli and the strain CICC32993 as Kodamaea ohmeri.

Gene expression profilinganalysis of mouse brain microvascular endothelial cells incubated with Cryptococcus neoformans

ZHONG Bin, LI Ping, NIE Shu, ZHU Hong-mei, WEN Hai

2014, 9(4): 203-206.

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Objective To dissect genomic changes in gene expression of mouse brain microvascular endothelial cells infected by Cryptococcus neoformans for providing new clues of fungal predilection for the central nervous system.Methods By comparing the gene expression profiles of bEnd.3 cells before and after infected by Cryptococcus neoformans of different serotypes, the changes of some important genes were found and then validated by RT-PCR.Results After comparison, we got 383 differentially expressed genes ,including 263 down-regulated and 120 up-regulated ones. And then we selected adhesion molecule gene CDH10 and selenium binding protein 1 gene SELENBP1 and validated their expression levels by RT-PCR . The results were in line with the microarray data.Conclusion Cryptococcus neoformans caused down-regulation of adhesion molecule CDH10 and up-regulation of SELENBP1 in brain microvascular endothelial cell might contribute to the yeast's invasion to host blood-brain barrier (BBB).

Mycological research of fungemia caused by Fusarium solani after multiple organ injury

KANG Yu-li, ZHU Jun-hao, LI Li, ZHAO Ying, ZHANG Qiang-qiang

2014, 9(4): 207-210,206.

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Objective To isolate and identify the phenotype of pathogenic fungus,to get molecular diagnosis and in vitro drug sensitivity of Fusarium in a patient with fungemia after multiple organs injury.Methods A young patient diagnosed as multiple organs injury, and his blood was collected from deep veins and cultured for both bacterium and fungi. The isolated fungus from the blood culture were identified further with phenotype and molecular methods. Finally, E-test was used to detect the antifungal susceptibility.Results The blood culture was negative for bacteria, but positive for fungi. The isolate was confirmed as "Fusarium solani" according to the morphology of the fungus and the Results of phenotypic and molecular identification. The isolate was sensitive to AMB and VOZ, but resistant to ITZ, CS and FCZ.Conclusion Fusarium could cause local and disseminated infections in humans, and it was always vital and negative to most of antifungal drugs. It is necessary to perform blood fungal culture for the patients who were still febrile even after the debridement and antibiotic therapy. The fungal strain could be identified by the phenotype and molecular analysis.

A comparative study of efficacy and cost-effectiveness of micafungin and itraconazole in the treatment of multiple myeloma with invasive fungal disease

JIN Li-na, FAN Rong, ZHANG Chun-yang, DU Juan, FU Wei-jun, XI Hao, FAN Jian-ling, LI Rong, ZENG Tian-mei, HOU Jian

2014, 9(4): 211-214.

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Objective In order to compare the efficacy and cost-effectiveness of micafungin and itraconazole in the treatment of multiple myeloma with invasive fungal disease.Methods We restrospectively analyzed the clinical data of 37 multiple myeloma patients with invasive fungal disease. Then we analyzed the efficacy of antifungal therapy, treatment time, costs, and the adverse reactions.Results Seventeen patients were treated with micafungin, and 20 patients were treated with itraconazole. The micafungin group had higher effective rate 76.46% (13/17) vs 70% (14/20),(P=0.42>0.05).In micafungin group,2 patients occurred gastrointestinal reaction. In itraconazole group, 3 patients occurred digestive discomfort reaction ,2 patients occurred hypokalemia and digestive discomfort reaction, 3 patients occurred drug-induced renal impairment, 1 patient's renal function recovered after the drug was stopped, other 2 patients turned up with acute renal damage. The micafungin group had a lower adverse reactions was 11.76% (2/17) vs 40% (8/20),(P<0.05).The C/Es were 263.73 and 289.11 in micafungin group, itraconazole group, respectively.Conclusions In the treatment of multiple myeloma with IFD, The effective rate of treatment between itraconazole and micafungin had no statistical difference but the adverse reactions had significant difference, the cost-effectiveness ratio of micafungin is lower than itraconazole.

A preliminary research of tetrandrine and fluconazole affecting the cell cycle in Candida albicans

LV Xia-lin, ZHANG Hong, SONG Yan-jun, LI Shui-xiu

2014, 9(4): 215-217.

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Objective To explore the effect of TET (tetrandrine) and fluconazole on the cell cycle in Candida albicans.Methods Flow Cytometry was used to measure the DNA content of each cell cycle phase of the FLC (Fluconazole) sensitive strain CA-1, which was exposed to TET, FLC and TET+FLC.The effect of TET (tetrandrine) and fluconazole on the cell cycle in Candida albicans was analysed by comparing DNA content of different cell cycle phases and calculating the PI% (proliferation inhibition rate).Results Compared with control group,TET and FLC could increase the DNA content of cell cycle in S-phase by 17.25% (P=0.018) and 6.54% (P>0.05) respectively,while the combination use of TET and FLC could markedly increase S-phase DNA content by 31.52% (P=0.002).It showed that TET can arrest C.albicans cells in S phase.Conclusions TET has potent effect of against Candida albicans by means of inhibiting DNA synthesis,arrest the cell cycle and inhibiting cell proliferation,while it would be more notable when combined with FLC.

In vitro antifungal susceptibility of terbinafine against 24 strains of black yeast like fungi

TAN Xin, DENG Shu-wen, GUO Yun, ZHANG Chao, ZHU Shuang, PAN Wei-hua, DENG Dan-qi, LIAO Wan-qing

2014, 9(4): 218-221.

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Objective To investigate the in vitro susceptibility testing of terbinafine against 24 strains of Black yeast like fungi.Methods CLSI M38-A2 guideline was applied. The media containing the fungi in the concentration of 0.5×10⁴-4×10⁴ CFU/mL were incubated at 30 for 5-7 days. The MIC and MEC values were observed.Results The tested 24 strain of black yeast like fungi were susceptible to terbinafine, MEC range:0.125 μg/mL-4 μg/mL,MEC₉₀:2 μg/mL,MEC₅₀:0.25 μg/mL,GM:0.392 9 μg/mL,Five strains had 100% growth inhibition,MIC range:1-4 μg/mL,MIC₅₀:2 μg/mL;MIC₉₀:4 μg/mL.Conclusion Terbinafine is active aganist black yeast like fungi in order Chaetothyriales. Most strains had MEC which 50% growth inhibition by terbinafine.

The identification of 94 clinical isolates from vulvovaginal candidiasis in Fujian area

CHEN Bo-rui, JIANG Ai-ping, XU Ming, LU Wan-ding, CAI Lei

2014, 9(4): 222-227.

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Objective To reclassify the 94 Candida strains from clinical vulvovaginal Candidiasis patients in Fujian area, and to evaluate the accuracy of CHROM agar identification by comparing with the API test and molecular biological analysis.Methods 1Identify the 94 strains by CHROM agar, and observe the microscopic morphology. 2Identify these strains by ITS sequence analysis of molecular biological method. 3Comparing the test reports from the clinical laboratory with the identification results of CHROM agar and ITS sequence analysis, the isolates in doubt are selected to have the API test and LSU sequence analysis.Results 1There were 78 strains identified as C.albicans, 10 strains as C.glabrata, 3 strains as C.parasilosis, 1 strain as Saccharomyces cerevisiae, and 1 strains as both C.glabrata and C.parasilosis. 2The CHROM agar could nearly correctly identify Candida, however, lacks specificity when it comes to rare isolates (ex.Saccharomyces cerevisiae). 3According to the comparing results, we could conclude that the identification of clinical laboratory just by the methods of simple morphology and CHROM agar test still has the error rate (10/94). 4The identification of molecular biological method had high accuracy rate. Besides, we could identify the C.parapsilosis complex was consist of the species of C.parapsilosis sensu strico,Candida metapsilosis and Candida orthopsilosis by this way.Conclusions The most part of clinical isolates of vulvovaginal candidiasis in Fujian area were still C.albicans, but the non C.albicans also occupied a certain proportion (16/94). Our result showed that the molecular biological methods were more accurate than the CHROM agar test in identification.

Trend of global mycology research based on bibliometric study (2004-2013)

JIA Ze-jun, YIN Cha, DENG Xiao-qun

2014, 9(4): 228-230.

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Objective To analyze developing trend of global mycology research.Methods The global and China mycology research-related publications were retrieved from the Web of Science database from 2004 to 2013, and Excel 2007, Matlab softwares were adopted to analyze the searching results quantitatively, including the cited frequency, h-index, fund paper ratio, high contribution organization etc.Results Global mycology research is on the rise in recent years, the United States has the absolute leading, China, Germany and Japan are also importmant contribution to the research. The total Chinese literature on mycology research ranked the 2th, with main contribution since 2007 (98.79%). Chinese universities are the main research institutions; the main sustentation fund were from the National Natural Science Foundation of China.Conlusion Since 2007 Chinese mycology research is in the good development opportunity, but still research level remains to be improved.

A case of fixed cutaneous sporotrichosis treated by cryotherapy combined with potassium iodide

ZHU Chen-yu, SUN Qiu-ning, LI Yun, FANG Kai

2014, 9(4): 235-236,230.

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This is a report of nasal fixed cutaneous sporotrichosis.A 51-year-old female farmer with 2-months history of verrucous hyperplasia on her nasal tip with exudation and crusts.It was diagnosed as fixed cutaneous sporotrichosis by the clinical manifestation, histopathological presentations,microscopic examination and fungal culture of sporotrichosis.After three months of cryotherapy combined with potassium iodide, the lesion disappeared.

Clinical analysis of 33 cases of tinea capitis in children

GAO Yu, QIAN Gui-ping

2014, 9(4): 237-240.

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Objective To observe epidemiology, transmission, pathogens and treatment effects of tinea capitis in children in Jiangyan areas.Methods In August 2006-May 2013,33 cases of tinea capitis in children were analyzed for diagnosis and treatment.Results In the 33 cases of tinea capitis in children,there were 24 cases of white ringworm,5 cases of black ringworm and 4 case of kerions,19 males and 14 females.The pathogenic fungi were Microsporum canis (22 cases),Trichophyton violaceum (2 cases),Trichophyton mentagrophytes (3 cases),Trichophyton tonsurans (3 cases),Trichophyton rubrum (1 case),Microsporum gypseum (2 cases). Patients with animals contact history accounted for to 42.42% (14/33).Administrated with terbinafine or itraconazole orally for 6 to 8 weeks, daily washing hair with sulfur soap,using ketoconazole naftifine cream topically,patients all got satisfied curative effect.The cure rate were 87.5% and 88.24%, the fungal clearance rates were 100% and 94.12% respectively at 8 weeks.Conclusion In recent years, the reason for the increase of the disease is related to feed cats, dogs and other pets, the main pathogens were Microsporum canis dear animal.The comprehensive treatment, curative effect is satisfied.

Progress on the transcription factors associated with azole resistance in Candida albicans

WANG Ying, LIU Jin-yan, SHI Ce, XIANG Ming-jie

2014, 9(4): 241-244,240.

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Lately, the rate of Candida albicans infection kept increasing, more and more resistant strains had been detected with the wide use of azoles. Sogreat challenge had been brought to clinical treatment. Some molecular mechanisms associated with azole resistance had been described, including the point mutation ,overexpression of target enzyme, increased expression of efflux pump and the formation of biofilm. Since the transcription factors could regulate the expression of drug resistance genes, more and more attention has been paid to the contribution of zinc cluster transcription factors to drug resistance, such as TAC1, MRR1, MRR2, UPC2, NDT80, and so on. Lots of researches revealed that the point mutations of transcription factors could cause overexpression of their regulated resistance genes and finally led to drug resistance. The review focused on the research progress of the transcription factors.

P2X7 receptor and inflammation cytokine

XI De-e, HAN Li,LU Jian-ming

2014, 9(4): 252-256.

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P2X7 receptor is a special subtype of purine receptors, it functions as the ion channel which controlled by ATP. It has high expression on monocytes, macrophages and neutrophils. Its activation will result in Ca²⁺ inflow, K⁺ outflow, non-selective membrane channel formation, and will launch a series of signaling pathway including NALP3 formation, inflammatory cytokines transcription activated by NF-κB through mitogen-activated protein kinase, the production of ROS and nitrogen medium, and will mediate the release of many inflammatory cytokines such as IL-1β, IL-6, IL-18, TNF-α, MIP-2, CCL2 and HMGB1, and will take part in the development of inflammation, which is closely related to inflammatory disease including alzheimer's disease, rheumatoid arthritis and asthma and fungal infection.

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