Chinese Journal of Mycology

Effects of SakA on the drug stress and virulence in Talaromyces marneffei

NING Xin-qiang, WEI Jin-ying, ZHENG Yan-qing, LIANG Hao, CAO Cun-wei

2018, 13(2): 65-70.

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Objective Environmental stress signals are transmitted by evolutionarily conserved MAPKs in most fungi, such as Hog1 in the budding yeast Saccharomyces cerevisiae, Spc1 in the fission yeast Schizosaccharomyces pombe and SakA in the Aspergillus nidulans. It has been shown that in Talaromyces marneffei, a pathogenic dimorphic fungus, the SakA (HOG1) gene is involved, not only in tolerance against oxidative and heat stresses, but also in playing a role in growth, morphology, asexual development and the integrity of the cell wall. In this study, the role of the TM SakA gene on the drug Stress and virulence were investigated. Methods We constructed a SakA knockout strain (ΔsakA) of TM to observe the co-culture of spores with macrophages and compare the growth status of wild plants and ΔsakA at drug pressures. In addition, we infected mice with wild and knockout strains, respectively, and the mortality and fungi loading were compared between these two groups. Results After being phagocytosed by macrophages for 24 h, the sausage-like yeast cells were observed in the wild-type strain while the ΔsakA was still in the form of spores and could not be transformed into yeast cells. The number of colonies formed by ΔsakA co-cultivation was less than that of the wild-type strain. Compared with the wild strain, ΔsakA is more sensitive to echinodermycin drugs. In the infected mouse model, the mortality and fungi loading of ΔsakA-infected mice were significantly lower than those of wild strains. Conclusion SakA gene plays an important role in the pathogenesis of TM and is associated with why the wild strains of TM is not sensitive to echinodermycin.

The research of quantitative real-time PCR testing different specimen in diagnosis of talaromycosis

ZHENG Yan-qing, SHI Na-na, CAO Cun-wei

2018, 13(2): 71-74.

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Objective To evaluate real-time fluorescence quantitative PCR (qPCR) in determining the load of Talaromyces marneffei in different samples of talaromycosis marneffei (TSM). Methods The loads of Talaromyces marneffei in pathological slices, biopsy tissues from suspected patients of TSM were measured by means of qPCR. Results The average load of Talaromyces marneffei in pathological slices was 3.01×10⁴ copies (1.07×10⁴~7.26×10⁴ copies), the load of Talaromyces marneffei in biopsy tissues was 4.68×10⁵ copies. Conclusion This study showed that qPCR determining the load of Talaromyces marneffei in pathological slices and biopsy tissues could be a useful method in TSM diagnosis.

Construction and identification of ORF19.6012-MYC strain in Candida albicans

LU Ren-yi, ZHANG Xiao-long, YAN Lan, JIANG Yuan-ying

2018, 13(2): 75-78.

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Objective To construct ORF19.6012-MYC strain in Candida albicans. Methods The recombinant fragment composed of the upstream, downstream of the ORF19.6012 and the MYC-SAT1 flipper was constructed by In-Fusion kit. Then the fragment was homologously recombined into the genomic DNA of SN152 by lithium acetate transfection. The MYC tag was added to the C terminus of the open reading frame of ORF19.6012. PCR and Western blotting were used to verify the positive strain. Finally, the growth curve and hyphal formation ability of the positive strain were determined. Results ORF19.6012-MYC fusion strain was successfully constructed. The growth and proliferation ability of ORF19.6012-MYC strain and the ability of hyphal formation were consistent with those of the parental strain. Conclusion The ORF19.6012-MYC strain was successfully constructed and would be used in the following experiments.

Susceptibility of Candida albicans to 5 antifungal agents: comparison of yeast and hyphal forms

LI Rong-yu, CHEN Jing, OU Cheng-ju

2018, 13(2): 79-82.

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Objective To compare the susceptibility of yeast and hyphal forms of Candida albicans to commonly used antifungal agents, so that to provide theoretical guidance for the pathogenic mycology research and clinical medication. Methods Susceptibility of 220 clinical isolates of Candida albicans to antifungal agents was determined using ATB Fungus 3 Kits (bioMerieux, France). Results The susceptibility rates of yeast and hyphal forms to amphotericin B, fluconazole, itraconazole, voriconazole and 5-fluorocytosine were 97.27% vs 97.73%, 81.82% vs 85%, 87.27% vs 90.46%, 92.72% vs 95%, 96.82% vs 97.27% respectively. MICs to fluconazole, itraconazole, voriconazole of yeast forms were higher than those of hyphal forms (P<0.05), No satistically difference was found to amphotericin B (P>0.05).The overall sensitivity of two tested forms to 5 antifungal agents was 85.46% (188/220). 32 yeast strains showed resistance and intermediation, but hyphal strains showed intermediation and sensitivity. Conclusion There were differences between the susceptibility of yeast and hyphal forms of Candidia albicans to antifungal agents. The antifungal activity of hyphal-form was higher than that of yeast-form.

Clinical study of domestic amphotericin B or in combination with other antifungal drugs therapy 54 cases of malignant hematologic patients with invasive fungal disease

LIU Jing, JI Hai-long, ZHAO Fang, ZHANG Rui, YIN Feng-lei, CAO Ting-ting, MA Hong-yu, XU Wei-xing, WANG Juan

2018, 13(2): 83-86,90.

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Objective To evaluate the efficacy and safety of domestic amphotericin B or in combination with other antifungal drugs in malignant hematologic patients with invasive fungal disease (IFD). Methods A retrospective clinical analysis was performed based on 54 patients with hematologic malignancies who firstly infected IFD were treated by amphotericin B or combinated with other antifungal drugs during April 2013 and April 2016 in our department. Results Overall response rate was 88.9%. The response rates of amphotericin B, amphotericin B combined with voriconazole, amphotericin B combined with caspofungin were 76.2% (16/21 cases),96.4% (27/28 cases),100% (5/5 cases). The main risk factors of the patients were the recovery of neutrophils and smoking history (P<0.05). The common lung CT findings of the patients were patchy ground glass shadow 63.0% (34/54 cases), consolidation 11.1% (6/54 cases), nodule 14.8% (8/54 cases). The major adverse reactions of the patients were hypokalemia, liver damage. Conclusion The domestic amphotericin B for malignant hematologic patients with IFD are economic, safe and effective antifungal drugs.If it is ineffective,then combined with other antifungal drugs can enhance curative effect. Although increased incidence of adverse reactions, but they can be controlled, so as to achieve more aggressive antifungal treatment, improve the treatment effectiveness and reduce the case fatality rate.

The analysis of pathogens and clinical features of 131 cases of children' superficial mycosis

HAI Le, MA Yue-e, SHI Yu-ling

2018, 13(2): 87-90.

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Objective To investigate the prevalence and pathogenic fungal species of superficial mycoses in children in our region. Methods Culture and fungal identification were performed on clinical specimens collected from 131 children with superficial mycoses in our hospital from Jan.2015 to Jan.2016. Results In these 131 cases, superficial mycoses on foot were most (47 cases, 35.9%), followed by superficial mycoses in the body trunk (32 cases, 24.4%), and there were the other superficial mycoses, onychomycosis (24 cases, 18.3%), in femoral region (12 cases, 9.2%), on hand (9 cases, 6.9%), on head (7 cases, 5.3%), respectively; boy, 84 cases (64.1%), girl, 47 cases (35.9%); the age distribution of boy was between eight months to 14 years old, and the girl's was 3 months to 14 years old; among the 96 isolated pathogenic fungi strains, the most common species were Trichophyton rubrum (37 strains, 38.5%), followed by Candid albicans (28 strains, 41.7%) and Microsporum canis (12 strains, 12.5%). After above, there were 3 Malassezia strains, 2 Candida tropicalis strains, other 10 Candida strains, 1 Epidermophyton floccosum strain, 1 Trichophyton mentagrophytes strain, and 2 strains of unclassified fungi. Conclusions In 131 cases of children's superficial mycosis, tinea pedis was the most common clinical type; the major pathogenic fungi were Trichophyton rubrum and Candida albicans.

Progess of topical antifungal agents for the treatment of onychomycosis

WANG Ruo-jun, LI Ruo-yu, WANG Ai-ping

2018, 13(2): 114-119.

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In this article, we reviewed topical antifungal agents for the treatment of onychomycosis at present, including the widely used agents, 8% ciclopirox nail lacquer and 5% amorolfine nail lacquer, and the agents recently approved by the FDA, 10% efinaconazole and 5% tavaborole solution, as well as luliconazole and tazarotene which being still in studies. The use of new formulations of topical antifungal drugs were also included in this review.

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