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    28 April 2016, Volume 11 Issue 2
    论文
    Transcription factor Pho4 is involved in the regulation of drug susceptibility of Candida albicans
    QIN Yu-lin, XU Hong-tao, ZHANG Lu-lu, ZHANG Jin-yu, JIANG Yuan-ying, CAO Yong-bing
    2016, 11(2):  65-69. 
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    Objective To study the influence of Candida albicans transcription factors (TFs) on drug susceptibility and the relevant regulating mechanisms, a screening of a TF deletion mutant library had been conducted.Methods The broth microdilution method and spot assay were used to determine the susceptibility of TF deletion strains.The expression of multidrug resistance genes MDR1, CDR1 and ERG11 were examined by real-time PCR, and the analysis of rhodamine 6G efflux was performed to evaluate the efflux pump activity.Results The measurement of drug minimal inhibitory concentration and spot assay indicated an increase of susceptibility to fluconazole and miconazole in PHO4 deleted mutant compared to the parental strain.Although the mutant had a higher expression of the drug resistance genes, the amount of rhodamine 6G efflux and the oxidative stress resistance of it were lower than the parental strain.Conclusion Deletion of PHO4 in C.albicans might cause a decrease in oxidative stress resistance and drug efflux ability, finally result in hypersusceptibility to antifungal drugs.

    The amplification and targeting vector construction of a polyketide synthases gene of Fonsecaea monophora
    XIAO Xing, ZHANG Jun-min, FENG Jiao, CHEN Zhi-wen, CHEN Chun-mei, HE Ya, HE Dan, SUN Jiu-feng, XI Li-yan
    2016, 11(2):  70-74. 
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    Objective To amplify and sequence a polyketide synthases gene of Fonsecaea monophora, and to construct its targeting vector.Methods A polyketide synthases gene of Fonsecaea monophora was amplified and sequenced.Then we designed primers to amplify the 5' and 3' flanking region of PKS gene from Fonsecaea monophora genomic DNA, and the Hygromycin B resistance gene (hyg) from plasmid pBHt1.At last, all fragments were inserted into the vector PDHt/sk.Result The sequence of PKS gene was acquired, and its length was 5 389 bp.The targeting vector of PKS gene was constructed, and the vector was identified by restriction enzyme digestion and nucleotide sequencing.Conclusion This study acquired the targeting vector of PKS gene of Fonsecaea monophora successfully, and laid a good foundation for biological functions research of PKS gene and melanin.

    Pterostilbene induces apoptosis in Candida albicans
    HU Dan-dan, BI Shuang, JIANG Yuan-ying, WANG Yan
    2016, 11(2):  75-78. 
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    Objective To explore the activity of Pterostilbene (PTE) in inducing apoptosis of Candida albicans.Methods We used apoptosis, necrosis and overall viability assays, caspase activity determination, ROS measurement and mitochondrial membrane potential assay to evaluate the activity of PTE in inducing apoptosis of C.albicans.Results After C.albicans treated with ≥4 μg/ml PTE, the percentage of apoptosis cells and activity of Caspase notably increased.Moreover, the mitochondrial membrane potential decreased significantly and the intracellular ROS increased.Conclusions PTE has an apoptosis-inducing effect on C.albicans, and this effect may be associated with ROS accumulation and mitochondrial injure.

    A loop-mediated isothermal amplification method for molecular detection of Histoplasma capsulatum
    LI Juan, CHEN Min, PAN Bo, LEI Wen-zhi, FANG Wen-jie, LIU Jia, HONG Nan, LI Ying-fang, LIAO Wan-qing, PAN Wei-hua
    2016, 11(2):  79-84. 
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    Objective To develop a molecular diagnostic approach for the diagnosis of Histoplasma capsulatum based on the LAMP technology.Methods To establish and optimize a specific primer set of the LAMP technology targeting the sequence of M Antigen for rapid and molecular detection of Histoplasma capsulatum at species level.Results In our study, no cross-reaction was observed after 90 min of LAMP reaction, including DNA of the phylogenetically related species such as Coccidioides immitis, Sporothrix schenckii, Penicillium marneffei, etc.The limit-of-detection of the LAMP for genomic Histoplasma capsulatum DNA was approximately 5.3×10-12 g per run (approximately 160 genomic copies).Conclusion Our proposed LAMP assay could represent a potential molecular detection tool for rapid diagnosis of Histoplasma capsulatum as a point of care test to be used in clinics in the future.

    Comparision of different interpretation criterions of fractional inhibitory concentration for evaluating the in vitro combined effect of antifungal agents
    WU Wei-wei, WANG Qian, GONG Jie, WAN Zhe, WANG Ai-ping, LI Ruo-yu
    2016, 11(2):  85-89. 
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    Objective To determine the interaction pattern of ketoconazole combined with terbinafine or naftifine, and to evaluate the effects of different interpretation criterions of fractional inhibitory concentration index (FICI) for the in vitro combined effect of antifungal drugs.Methods According to CLSI M38-A2 and M27-A3, a checkerboard broth microdilution method was performed to investigate the in vitro activities of ketoconazole in combination with terbinafine or naftifine against 100 clinical strains isolated from patients with mycosis, including dermatophyte, Candida and Malassezia.Different criterion for the interpretation of fractional inhibitory concentration were employed to evaluate the in vitro combined effect of ketoconazole in combination with terbinafine or naftifine.Standard A:synergy is defined by FICI ≤0.5, antagonism by FICI>4, and no interaction by FICI>0.5, but ≤4;Standard B:synergy is defined by FICI ≤0.5, additive by FICI>0.5, but ≤1, no interaction by FICI>1, but ≤2, and antagonism by FICI>2;Standard C:synergy is defined by FICI ≤1, additive by FICI=1, no interaction by FICI>1, but ≤2, and antagonism by FICI>2.Results Standard A had good consistency with Standard B, Standard C is relatively lenient.According to Standard C, the interpretation might significantly expand the range of synergy.Conclusions The interpretation criterion of fractional inhibitory concentration (Synergy is defined by FICI ≤0.5, antagonism by FICI>4, and no interaction by FICI>0.5, but ≤4) has the advantage of simplicity, convenience and good consistence, and it is worthy of clinical application.

    In vitro antifungal spectrum and activity of 1% luliconazole cream compared with 2% ketoconazole cream, 1% terbinafine cream and 1% naftifine-0.25% ketoconazole cream
    FENG Xiao-wei, HU Wen-ying, RAN Yu-ping, ZHUANG Kai-wen, DAI Ya-ling
    2016, 11(2):  90-94. 
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    Objective To research the anti-fungal spectrum and activity of the 1% luliconazole cream compared with other three creams that contain of 2% ketoconazole, of 1% terbinafine, of 1% naftifine -0.25% ketoconazole cream respectively.Methods Thirty-two isolates of pathogenic fungi belonging to 14 species isolated from clinic and three species of Malassezia standard stains were enrolled into the experiment.The agar diffusion method was used to judge drug sensitivity.Organism suspension of each species was spread on the surface of the plate of the optimal media containing 2% agar.Then wells were made in the plate and four types of cream were put in each well respectively.After seven-day incubation, the diameter of the inhibition zone around the well full of each cream was observed and recorded.Results The inhibition zone around the well filled with 1% luliconazole cream for all experiment isolates was observed, with the mean diameter of 50.35 mm.Similarly, the mean diameter of inhibition zone of 1% naftifine -0.25% ketoconazole cream for all experiment isolates was 52.46 mm.About 1% terbinafine cream, the mean diameter was 49.24 mm but there was no inhibition zone observed around Candida spp.and Fusarium solani.As for 2% ketoconazole cream, the mean diameter was 33.22 mm but there was no inhibition zone observed around Fusarium solani.There were no significant significances for mean diameters of the inhibition zone when comparing 1% luliconazole cream with 1% naftifine -0.25% ketoconazole cream, but when comparing 1% luliconazole cream with 2% ketoconazole cream and 1% terbinafine cream, the difference was statistically significant (P=0.000).Conclusion The anti-fungal spectrum and activity of 1% luliconazole cream are nearly the same as that of 1% naftifine -0.25% ketoconazole cream, and is stronger than that of 2% ketoconazole cream and 1% terbinafine cream.

    Clinical analysis of 16 cases of tinea of vellus hair in children
    YAN Wei, FU Mei-hua, MEI Huan, CHEN Wei, LIU Wei-da
    2016, 11(2):  95-98. 
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    Objective To investigate the 16 cases of tinea faciei of vellus hair in children, to explore the related factors of the incidence, common pathogenic fungi and effective treatment.Methods We collected 16 patients with tinea faciei of vellus hair from April 2014 to July 2015 and analyzed their basic informations, medicine histories and clinical characteristics and animal contact histories.The specimens of 16 cases were cultured and identificated.All the patients were given terbinafine cream or bifonazole gel and oral terbinafine combined treatment.Results Of the 16 patients, there were 15 cases of animal contact history, 14 cases of previous treatment history.Fifteen strains of pathogens were isolated, including 13 Trichophyton mentagrophytes, 1 Trichophyton violaceum and 1 Microsporium canis.All the patients recovered well after treatment.Conclusion Contacting with infected pets was the significant risk factor to children with tinea faciei of vellus hair.The most commenly pathogenic fungus was Trichophyton mentagrophytes.Timely and effective mycological examination is helpful to diagnosis tinea facial of vellus hair.

    The clinical characteristics of cryptococcal meningitis in apparently immunocompetent patients
    LIU Jia, FANG Wen-jie, HONG Nan, LI Juan, CHEN Min, PAN Wei-hua, LIAO Wan-qing
    2016, 11(2):  99-102. 
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    Objective To analyze the clinical features of cryptococcal meningitis (CM) in apparently immunocompetent patients.Methods Clinical data of 44 CM cases in immunocompetent patients were collected and analyzed in Shanghai Changzheng Hospital from 2000 to 2015.Results Headache, nausea, and fever were the most common leading symptoms of the patients whose mean age was 35.9 years old.The difference in the male-to-female gender ratio of immunocompetent patients was not significant in this survey.And the mean pressure of CSF was (339.3±75.0) mmH2O.Cryptococcus neoformans was positively detected using Indian ink stain in 41 patients (97.6%, 41/42), followed by CSF cryptococcal antigen latex agglutination test 33 patients, CSF culture was positive in 11 patients.Six patients were given AmB and flucytosine, which was the most common induction therapy.The prognosis was somewhat optimistic of our study, resulting only one patient with mild paralysis.Conclusions Cryptococcal meningitis also can occur in immunocompetent hosts without any apparent underling disease, and the clinical manifestations may be not so typical at early stage, which can lead to misdiagnosis and missed diagnosis.To achieve higher survival and lower mortality rate, relevant CSF examinations are recommended for those suspected patients.

    Retrospective analysis of primary cutaneous mucormycosis in China: a review of 32 cases
    LV Hong-li, LI Cheng, LI Yan, TAN Juan, XU Ren-chao, TANG Jin-hui
    2016, 11(2):  103-107. 
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    Objective To investigate and discuss the epidemiology, risk factors, clinical manifestations, pathomycete and treatment strategies for primary cutaneous mucormycosis.Methods Thirty-two cases of primary cutaneous mucormycosis were searched from the electrical database, and all the clinical information was collected and evaluated.Results Twenty-seven articles (32 cases) were included in the study.Median age was 39.97 years old, with a male predominance (M:F=9:7).The major clinical manifestations were local erythema, papula and nodule, which might spread into infiltrating plaque associated with ulceration and necrosis.Diagnosis was mainly dependent on mycologic and/or histopathological examination.Amphotericin B was the first-line antifungal drug for the disease.Conclusion Primary cutaneous mucormycosis is a rare severe devastatind deep fungal infection.Enhancing the understanding of its clinical characterisctics would contribute to the early diagnosis and timely treatment, and amphotericin B was the first-line choice.

    Epidemiological investigation of tinea pedis in naval soldiers coming from different stations in recent 3 years
    LIU Qi-fang, SUN Tao, FANG Wei, YANG Run-ping, LI Xiao-peng
    2016, 11(2):  108-110. 
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    Objective To analyze the prevalence of tinea pedis of naval soldiers coming from different stations in recent 3 years.Methods From 2013 to 2015, survey was preformed yearly to investigate theincidence rate, clinical type and pathogenic fungi of tinea pedis of 600 naval soldiers selected randomly from 4 stations including Beijing, Dalian, Ningbo, Sanya.Results Only in 2013 there was a statistical significant difference in the prevalence of tinea pedis of naval soldiers between 4 stations (P<0.05), the incidence rate of tinea pedis in Sanya station is 18.0%, higher than other 3 stations.In recent 3 years the incidence rateof tinea pedis are elevated annually in Beijing, Dalian stations (P<0.05).In 4 stations, Trichophyton rubrum was themost common pathogenic fungi followed by Trichophyton mentagrophytes, Microsporum canis, Candida albicans, Epidermophyton floccosum.In Sanya, Ningbo stations the detection rates of Trichophyton rubrum were respectively 70.1%, 70.9%, higher than in Beijing, Dalian stations which were respectively 62.8%, 56.5% (P<0.05);but the detection rates of Trichophyton mentagrophytes were respectively 12.9%, 16.1%, lower than in Beijing, Dalian stations which were respectively 26.4%, 26.3% (P<0.05).In Sanya stations the detection rate of Epidermophyton floccosum was the highest among all stations (P<0.05), accounting for 1.9%.In 4 stations there was not statistical significance difference in the constituent ratios of blister-scale tinea pedis.But in Sanya station the constituent ratio of infusion-erosion tinea pedis is 23.0%, which was the highest among all stations (P<0.05);and the constituent ratio of hyperkeratotic cases was the lowest (P<0.05), accounting for 20.3%.Conclusion In recent 3 years the incidence rate of tinea pedis of naval soldiers were elevated annually in Beijing, Dalian stations, and clinical type and pathogenic fungi of tinea pedis between 4 stations carried a significant difference.

    Tinea barbae due to Trichophyton interdigitale: a case report
    WANG Xiao-wen, LI Ya-li, WANG Hong, CHEN Wei, WAN Zhe, TU Ping, LI Ruo-yu, WANG Ai-ping
    2016, 11(2):  113-116. 
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    A 31-year-old goat farmer presented with aggravating lesions on his upper lip, lower jaw, cheek and the front of the neck for 4 months.Physical examination showed red swelling plaques, with marked nodules, pustules, pyorrhea and crusts on them and obvious tenderness.Most beards were off, with a few broken beards left and easy to pull out.Direct microscopic examination of pus showed septate hyaline hyphae with branches.Trichophyton interdigitale was isolated and identified from pus cultures.And histopathological features showed granulomatous changes.The diagnosis of tinea barbae caused by T.interdigitale was made based on the above findings.The patient was successfully treated with oral itraconazole 400 mg/d for 6 weeks, followed by the combination of oral itraconazole 200 mg/d and terbinafine 250 mg/d for another 6 weeks.

    Trichosporon mucoides isolated from the toes in a tinea pedis patient
    HU Wen-ying, XU Feng-ni, RAN Yu-ping, DAI Ya-ling, ZHUANG Kai-wen
    2016, 11(2):  117-118,112. 
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    Trichosporon mucoides isolated from the toes in a tinea pedis patient was reported.The patient had a history of itching and scaling of feet for 6 months.Her toes were sweaty with white scaling, and her big toe of right foot had plantar warts.Direct microscopic examination of scale samples showed septate hyaline hyphae;fungal colonies developed on Sabouraud dextrose agar after inoculating for 2 weeks were identified as Trichosporon mucoides by molecular identification.The diagnoses were tinea pedis and plantar warts.The lesions were completely cured by antifungals, cryotherapy and imiquimod cream.

    A case of cutaneous chromoblastomycosis caused by Fonsecaea monophora
    XIANG Yun, JI Qing, CAI Hong-wei, MA Ling-ling
    2016, 11(2):  119-121. 
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    To report a case of chromoblastomycosis caused by Fonsecaea monophora.A 72-year-old man was admitted to the hospital with a more than 20-year history of plaque on the right shoulder.Direct microscopic examination and fungal culture, Histopathologic examination, PCR and ribosomal internal transcribed spacer (ITS) sequence analysis were performed for diagnosis.The thick-walled, brown sclerotic bodies were observed by direct microscopic examination and histopathologic examination.Fungal culture yielded black brown villiform colonies.Rhinocladiella type and cladosporium type conidia were observed in the slide culture.The isolate was identified as F.monophora according to the gene sequence of internal transcribed spacer (ITS).The patient was cured after treatment with terbinafine (250 mg, qd) alone for 7 months.The case of cutaneous chromoblastomycosis caused by F.monophora was diagnosed by histopathology, mycology and molecular biology methods and terbinafine was effective to it.

    Reasonable application of scientific teaching theory in Medical Mycology Teaching
    FANG Wei, ZHU Yuan-jie, CHEN Min, YANG Ya-li, CHEN Jiang-han, LIAO Wan-qing
    2016, 11(2):  122-124. 
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    In recent years, with the increase of the incidence of fungal infections, teaching about Medical Mycology is particularly important.Guided by scientific teaching theory, instructional design should be produced in following fields such as the task of teaching, teaching objectives, teaching strategies and teaching evaluation, which has a certain significance for the teaching of Medical Mycology.

    The research progress on Candida albicans colonization and infection in the mice gastrointestinal tract
    ZHU Fang, HE Li-juan, HUANG Xin, AN Mao-mao, JIANG Yuan-ying
    2016, 11(2):  125-128. 
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    Candida albicans was regarded as a commensal within the gut in humans and the gut had been considered as an important site for Candida albicans invasive infection.Antibiotic(s)-treated mice orally given Candida albicans could closely mimic the clinical manifestations, including gastrointestinal colonization and invasive infection.Immunosuppressants were required for the worse invasive infection.There we summarized the progress on the models of Candida albicans colonization and/or invasive infection in the gastrointestinal tract from related literatures since 2000.

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