Table of Content

28 December 2013, Volume 8 Issue 6

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Establishment of a real-time fluorescence quantitative PCR method for the detection of clinical Exophiala dermatitidis

WANG Fang, MU Chao, ZHAO Jing-ya, GAO Li, HU Xiao-hua, HAN Li, TIAN Shu-guang

2013, 8(6):  321-324. 

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Objective To establish a novel fluorescent quantitative PCR (qPCR) method for the detection of Exophiala dermatitidis based on TaqMan probe technology.Methods Specific primers and fluorescent labeling probes were designed, according to the sequence of internal transcribed spacer (ITS) regions of E.dermatitis genomic sequence (GenBank:JN675373.1). And the condition of qPCR was optimized.E.dermatitis isolated from clinical specimens were tested as positive control, and other species of fungi and bacteria were used as negative control, to evaluate the detective effect of specificity, sensitivity and repeatability.Results In this study, primers and probes were designed to amplify Exophiala dermatitidis specific sequence, and Exophiala dermatitidis isolated from clinical specimens obtained obvious amplification curve in the reaction, whereas the 20 negative control strains, such as Exophiala spinous,Aspergillus fumigatus,Candida albicans,Cryptococcus neoformans,Penicillium marneffei,et al were not amplified in the CT value≤38 range. The qPCR exhibited high sensitivity (10 Cp/μL) and a good linearity from 10³ to 10⁷ copies (R²=1.000).Conclusion We successfully established a quantitative PCR method with high specificity, sensitivity, good repeatability for detection of E.dermatitis.The method could contribute to the early diagnosis of E.dermatitis infection and targeted therapy.

Up-regulation of miR-146a by Cryptococcus neoformans via Dectin-1

HUANG Hong-yu, LIU Jing-bo, WEI Yun-yu, YAN Zi-he, HAN Zhi-jun

2013, 8(6):  325-328. 

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Objective To investigate whether miR-146a involved in the immune response against Cryptococcus neoformans.Methods The relative expression of miR-146a in peripheral blood mononuclear cell of cryptococcal meningitis patients and healthy individuals was determined by RT-PCR. Heat-killed Cryptococcus and laminarin, a Dectin-1 blocker, were used to stimulate PBMCs from healthy individuals, miR-146a expression were analyzed.Results miR-146a was increased in the group of cryptococcal meningitis patients. Heat-killed Cryptococcus could increase miR-146a expression in PBMCs, and this effect could be partially inhibited by laminarin.Conclusions Cryptococcus up-regulate miR-146a expression via Dectin-1. Mir-146a was involved in the immune response against Cryptococcus and further study should be taken to identify its potential mechanisms.

In vitro interactions of 28 kinds of drugs with fluconazole against Candida albicans

WANG Li, MAO Wei-an, CAO Yong-bing, WU Jian-hua

2013, 8(6):  329-333. 

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Objective To screen the drugs which may have synergistic effects with fluconazole from 28 kinds of drugs including immunosuppressive agents, antilipemic agents, antidepressants, antibiotics and non-steroidal anti-inflammatory drugs.Methods 28 kinds of drugs within 5 main types were formulated intothe uniform concentrations of 100 μg/mL, then were dripped into the fungal suspensions of Candida albicans including fluconazole-resistant strains (100,103) and sensitive strains (sc5314,y0109), singly or together with fluconazole of low concentrations, respectively. Then the clarity of the fungal suspensions were observed respectively, the sensitive strains after 24 h, while the resistant strains after 48 h. "+" was noted for the completely clarified cases, while "-" for the turbid cases accordingly. The test was repeated three times simultaneously.Results Sertraline and other 6 kinds of drugs could inhibit the growth of all the Candida albicans under the concentration of 100 μg/mL singly, which had more significant effects when combined with fluconazole. Rifapentine and other 5 kinds of drugs could only inhibit the growth of the sensitive strains under that concentration singly, but were still effective on the resistant strains when combined with fluconazole of low concentrations.Conclusion Some drugs such as sertraline, rifapentine may have synergistic inhibition effect on Candida albicans with fluconazole.

The effect to growth of superficial fungi by exposure to germicidal UV light

LIU Li, LAI Wei

2013, 8(6):  334-337. 

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Objective To investigate the effect to growth of superficial fungi by exposure to germicidal UV light.Methods Laboratorial fungi consisted of Trichophyton rubrum,T.mentagrophytes,M.canis Bodin,E.floccosum,C.albicans,C.glabrata,C.parapsilosis,C.tropicalis,C.krusei,Penicillium spp.,Scopulariopsis spp.,Acremonium spp. were inoculated on SDA that were exposed to UV light. All the tested fungi were divided into groups by exposure power, exposure distance and exposure time and morphological variations of hyphae and spores were observed with transmission electron microscope.Results M.canis Bodin,E.floccosum,C.albicans,C.parapsilosis,C.krusei stoped growing under exposure to germicidal UV light while others managed to be alive.Conclusions Germicidal UV light has different disinfection to some common superficial fungi, the degree of the disinfection depended on exposure power, distance and time.

A case of cutaneous chromoblastomycosis caused by Fonsecaea monophora and laboratory study

WANG Run-chao, WAN Zhe, CHEN Wei, TU Ping, LI Ruoyu, WANG Ai-ping

2013, 8(6):  338-341. 

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Objective To report a case of chromoblastomycosis caused by Fonsecaea monophora and related laboratory investigation.Methods A 60-year-old woman was admitted with skin granulomatous lesions on the left hand for more than one year.Direct smear examination and fungal culture,Histopathologic examination, scanning electron microscope examination, PCR and ribosomal internal transcribed spacer(ITS)sequence analysis were performed for diagnosis.The antifungals susceptibility was tested according to M38-A2 published by CLSI.Results Sclerotic bodies were observed by direct smear examination.Histopat hologic examination showed chronic granuloma and round,thick-walled sclerotic bodies.Fungal culture produced olivaceous to black colonies.Rhinocladiella type and cladosporium type conidia were observed in the slide culture and scanning electron microscope examination. Ribosomal DNA ITS domains were amplified,and the product was sequenced, which was aligned with 100%sequential similarity to multiple sequences of F.monophora in the GenBank database. The MICs for itraconazole and terbinafine were 1.0 μg/mL and 0.015 6 μg/mL against this strain, After treatment with terbinafine (250 mg,QD) alone for 6 weeks, and itraconazole (200 mg, QD) combined with terbinafine (250 mg,QD)for 8 weeks,the patient was cured.Conclusions The case of cutaneous chromoblagtomycosis was caused by F.monophora identified by histopathology, mycology and molecular biology, Itraconazole with terbinafine has good effect.

Penicilliosis marneffei with non-HIV-infected：two cases report and literature review

YE Feng, LUO Qun, ZHOU Ying, XIE Jia-xing, GU Ying-ying, SU Dan-hong, CHEN Rong-chang

2013, 8(6):  342-347,377. 

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Objective To describe the clinical feature of penicilliosis marneffei and heighten the ratio of diagnosis and treatment.Methods Two cases of disseminated penicilliosis marneffei in immunocompetent hosts from this hospital were presented.Result Case 1: 37-year old, male, was admitted because of recurrent episodes of coughing and fever for one month and developed pain in both shoulders and dizziness. An enlarged left supraclavicular and inguinal lymph node were palpated. Cranial MRI showed intracalvarium and retropharyngeal abscess. Disseminated penicilliosis marneffei with secondary seizure was diagnosed by transbrochial lung biopsy (TBLB)and pus fungal culture. The patient received a treatment with amphotericin B liposome and recovered. Case 2: 32-year old, male, was admitted due to a productive cough for 5 months, subcutaneous mass with fever for 3 months. During the treatment with amphotericin B liposome intravenously patient developed septic shock and DIC.Conclusion As the first time in China, penicilliosis marneffei is reported to disseminate to central nervous system. TBLB and pus culture are useful for the the diagnosis. Amphotericin B liposome is effective for the recurrent disseminated Penicillium marneffei and early diagnosis is the key for the outcome of the patients with this treatment.

Retrospective review of 12 cases of penicilliosis marneffei

TANG Xu-hua, ZHUANG Niang-qiao, ZHOU Hui, HU Rong-xin, ZHANG Xing-qi

2013, 8(6):  348-351. 

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Objective To study the clinical characteristics and outcome of patients with penicilliosis marneffei.Methods Retrospective review of 12 inpatients with penicilliosis marneffei in our hospital during ten years from January 2003 to October 2012.Results In addition to previously reported clinical manifestations including fever, rash, cough, dyspnea, serous effusion, hepatosplenomegaly, lymphadenopathy, digestive disorder, and osteolysis, we also observed pharyngalgia, edema of lower limbs, and vascular stenosis or occlusion as well. Laboratory examinations indicated 6 patients had abnormal routine urine examination, of whom 4 had elevated serum creatinine. Three out of four HIV positive patients were response to itraconazole combined with or without amphotericin B. Five out of eight HIV negative patients were response to voriconazole combined with or without other antifungal agents, and the other 3 were dead 2 to 11 days after the initiation of antifungal agents. Two of the 3 dead presented with ascites and dyspnea.Conclusions Pharynx, urinary system, peripheral circulation can also be involved in penicilliosis marneffei in addition to mononuclear phagocyte system.Penicillium marneffei is sensitive to voriconazole and itraconazole in vivo. Ascites and dyspnea possibly suggests poor prognosis. Early diagnosis and treatment of penicilliosis marneffei could improve the outcome.

Effectiveness of itraconazole in the prophylaxis of fungal infections in AIDS patiens：a meta-analysis of randomized controlled trials

YANG Xiao-yi, JIANG Jun-jun, YE Li, HUANG Jie-gang, ZHOU Bo, LIANG Bing-yu, TAO Ren-chuan, CAO Cun-wei, SHAO Yi-ming, LIANG Hao

2013, 8(6):  352-357. 

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Objective To evaluate the effectiveness of itraconazole in the prophylaxis of fungal infections in AIDS patiens.Methods A computerized literature searching had been carried out in Cochrane Database of Clinical Controlled Trials, PubMed, EMbase, EBSCO, Wanfang and Tsinghua Tongfang database, from January 1981 to July 2013. All randomized controlled trial (RCT) papers about prophylactic use of itraconazole to prevent fungal infections in AIDS patiens were enrolled. Meta-analysis was conducted using Stata 10.0 to calculate the pooled RR value (95%CI). Consistency test was performed.Results Finally 3 pieces of RCT papers were enrolled. It showed that prophylactic use of itraconazole can significantly reduce the rate of fungal infections in AIDS patients, with the pooled relative risk (RR) and 95% confidence interval (CI) was 0.54 (95%CI:0.44~0.67),P<0.05. However it showed no statistically significant differences in the rate of invasive fungal infections, with the pooled relative risk (RR) and 95% confidence interval (CI) was 0.38 (95%CI:0.13~1.10),P >0.05. There was no statistically significant differences in case fatality rate, with the pooled relative risk (RR) and 95% confidence interval (CI) was 1.14 (95%CI:0.79~1.65),P >0.05.Conclusion Prophylactic use of itraconazole in AIDS patients can significantly reduce the overall rate of fungal infections, but for invasive fungal infections has not been shown. Whether AIDS patients should use itraconazole to prevent fungal infections, it needs to be assessed carefully, and generate different prevention strategies according to the rates of different disease stages.

A survey of airborne fungi and and correlation study with bronchial asthma from 2007 to 2011 in Ezhou

XIONG Han-zhong, YIN Liang-qiong, FU Da-dong, ZHANG Fu-yu, HUO Shu-ping, WANG Hao-yi, CHEN Kai-qi, WANG Zhi-an

2013, 8(6):  358-360. 

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Objective To discuss the relationship betweenin airborne fungi and bronchial asthma from May 2007 to April 2011 in Ezhou,in order to understand the bronchial asthma in this area.Methods Using survey mathods of sheet glass and culture plate of exposure determination to count the species and quantity of airborne fungi in Ezhou. Fungi allergen skin test was performed on 100 cases which were diagnosed bronchial asthma and find out that 35 patients showed positive results on the intradermal skin test, Serum specific IgE was given to 35 patientsahd 40 healthy people.Results 35 cases showed positive result in 100 cases of bronchial asthma patients in Intradermal group, The positive rate was 35%,while 31 cases showed positive in specific IgE assay, coincidence of detection rate was 88.58%, there were 2 cases showed positive reaction in healthy people, the positive rate was 5%, The difference in positive rate between the two groups was significant (P<0.05).Conclusion airborne fungi is closely related to bronchial asthma, and it is one of sensitinogen to cause bronchial asthma. And also need to pay attention to environmental health to reduce the number of airborne fungi.

Clinical observation of Compound Kushen lotion plus itraconazol capsule in the treatment of malassezia folliculitis

BIAN Xian-li, ZHOU Xiang-ling

2013, 8(6):  364-365,373. 

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Objective To observe the curative efect of Compound Kushen Lotion plus itraconazol capsule in the treatment of malassezia folliculitis.Methods 48 cases of patients with malassezia folliculitis were divided into treatment group (24 cases) and control group (24 cases).Both two groups were treated with oral itraconazol capsule 200 mg,once a day for 2 weeks;treatment group were treated with additional Compound Kushen Lotion (external application),twice a day for 4 weeks.All the patients observed the curative efect and retested the microscopic fungi 2 weeks and 8 weeks later from the first day they have medicine.Results After 2 week's treatment,the total efectiveness of treatment group (75.0%) was higher than control group (41.7%). The difference was statistically significant (χ²=5.49,P<0.05).At 8 weeks,the total efectiveness of treatment group (91.7%) was higher than control group (62.5%). The difference was statistically significant (χ²=5.78,P<0.05).Conclusion The curative effect of Compound Kushen Lotion plus itraconazol capsule in the treatment of patients with malassezia folliculitis is significant,and it can be generalized.

A study of the mycological knowledge in the Zhiwu Xue of the Late Qing Dynasty

LU Di

2013, 8(6):  366-368. 

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As the first Chinese book introducing modern Western botany, the Zhiwu Xue written by Li Shanlan of the late Qing dynasty contains a small amount of mycological knowledge that enlightened the development of modern Chinese mycology. Based on the analysis of this part, it is found that fungi are taken as a group of plants, which is called "Xun Lei" and belong to "Tong Zhang Lei" (lower plants). However, compared with the creative translations of botanical terms, the mycological terms in the Zhiwu Xue were translated randomly, and there are mistakes in the relevant text. This book mentions the notions of the modern Chinese mycological terms, such as "Zhenjun", "Baozi" and "Zinang", but doesn't invent the corresponding Chinese terms. The reasons for this situation are closely related to the inferior status of mycology (mycology was undeveloped, and the proportion of mycological knowledge in botany books was small) then in addition to the problems of the translators and times.

Advances in the application of Matrix-assisted laser desorption ionization time-of-flight mass spectrometry for identification of clinically pathogenic fungi

WANG Chong, ZHAN Ping, LIU Wei-da

2013, 8(6):  369-373. 

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Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS)-based systems is a rapid and reliable method in identification of microorganism.With the improvement of identifiable fungal species spectrum,the application of MALDI-TOF MS systems have been introduced to identification of clinically pathogenic yeasts,yeast-like fungi,filamentous fungi and dermatophytes.

Advances in antifungal effect of sertraline

CAO Wen-juan, LUO Zhi-cheng

2013, 8(6):  378-380,384. 

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The incidence of fungal infections is rising worldwide. Along with the widespread use of antifungal agents, drug resistance is becoming more prominent. However, effective and safe antifungal agents are very scarce in clinics. Therefore, looking for new antifungal agents is a hot focus of researching works at present. Sertraline is a selective 5-serotonin reuptake inhibitors, commonly used in depression, obsessive-compulsive disorder, premenstrual syndrome, and so on. Ten years ago, a few of scholars found that sertraline has a certain inhibitory effect on the vitality of Candida and Aspergillus, and carried on the preliminary discussion about the antifungal mechanism. According to collecting and reading literature at home and abroad, this paper summarize the antifungal effects of sertraline.