Chinese Journal of Mycology

Preliminary screening active constituent against Candida albicans resistant to fluconazole from Rubus chingii Hu fruits

HAN Bing, LIN Hui, CHEN Jia, YU Yi-Qun, CAO Yong-Bing, JIANG Yuan-Ying

2012, 7(6): 326-329.

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Objective To compare three methods (cold-soak method, ultrasonic wave method and hot reflux method) of extracting antifungal constituents from Rubus chingii Hu fruits and preliminary screen active part of the extracts.Methods The three extractive methods were evaluated through antifungal activity in vitro of extracts from Rubus chingii Hu fruits combining with fluconazole against durg-resistant Candida albicans and extraction rate. The compositions of these extracts were separated with petroleum ether, chloroform, ethyl acetate, n-butyl alcohol and water. Screening out the active part were performed through evaluating antifungal activity in vitro of parts from the extracts of Rubus chingii Hu fruits combining with fluconazole against durg-resistant Candida albicans.Results The extraction rate of the Cold-soak method, ultrasonic wave method and hot reflux method was 21.0%, 16.8% and 12.8%. The FICIs of the extracts using three methods combining with fluconazole against Candida albicans 100 were 0.035 2, 0.032 2 and 0.046 9, and 0.033 2, 0.031 7 and 0.039 1 against Candida albicans 103. The FICIs of petroleum ether, chloroform, ethyl acetate, n-butyl alcohol and water part combining with fluconazole against Candida albicans 100 were ＞1, 0.502 0, 0.507 8, 0.033 2 and ＞1, and ＞1, 0.531 3, 0.507 8, 0.033 2 and ＞1 against Candida albicans 103.Conclusions Ultrasonic wave method is simple, reliable and efficient. Compared with the other methods, it is the best method of extracting antifungal constituents from Rubus chingii Hu fruits. The active part of the extracts from Rubus chingii Hu fruits is the n-butyl alcohol part.

Rapid identification of clinical important Aspergillus using real-time PCR combined with melting curve analysis

SHI Jun-yan, GUO Li-na, XU Ying-chun, WANG Yao, WANG He, LIU Wen-jing

2012, 7(6): 330-334,338.

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Objective Real-time PCR combined with melting curve analysis were carried out for rapid identification of clinical important Aspergillus to the species level.Method (1) PCR amplification of fungal ITS region, and then sequence alignment were done to identificate Aspergillus isolates correctly and design species specfic primers and probes. (2) Five species of clinical important Aspergillus were identified to species level according to different melting temperatures (Tm) of species specific probes in a biprobe real-time PCR.Results (1) The Tm value of A.terrus specific probe were 64.5℃ and 55.2℃, and that of A.flavus,A.fumigatus,A.nidulans and A.niger specific probe was 57.4℃,61.4℃,65.8℃,and 67.7℃ respectively.Phialophora verrucosa and Rhizopus microspores had cross reactions with A.fumigatus specific probe.(2)The detection limit of A.fumigatus was 56.8 fg, and that of A.flavus 1.11×10³ fg,A.niger 13.7 fg,A.terrus 1.23×10² fg and A.nidulans 7.80×10² fg.(3)The Tm value fluctuation was within 0.5℃ in repeated reactions.Conclusion The real-time PCR is time-saving and with high sensitivity, specificity and repeatablity. Therefore, it is helpful for diagnosis of invasive aspergillosis and the use of antifungal drugs.

The relationship between AQP4 and brain edema of cryptococcal meningitis model in mice

FAN Yi-bin, WEN Hai, ZHU Hong-mei, XU Hong, ZHAO Jin

2012, 7(6): 335-338.

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Objective To investigate the relationship between AQP4 and brain edema in murine model with central nervous system cryptococcal infection.Methods Mice were randomLy devided into two groups.Test group mice were injected with Cryptococcus neoformans via the tail vein,while control group mice received saline. Western Blotting technology was used in 6 h, 12 h, 24 h, 48 h, 72 h to detected AQP4 protein expression changes in the brain tissue of mice.Results Comared with the control group, AQP4 protein expression in the test group began to rise at 24 h and reached maximum at 48 h.Conclusions AQP4 play protective role in cerebral edema caused by Cryptococcal meningitis.

Fermentation culture of Cryptococcus neoformans benefiting for purification of mannoprotein

LI Ping, TAN Hong-yue, ZHONG Bin, CHEN Li-hua, HU Chan, ZHU Hong-mei, WEN Hai

2012, 7(6): 339-341,361.

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Objective To study the possibility of fermentation culture of Cryptococcus neoformans and the advantages of fermentation culture supernatant used for purificating mannoprotein.Methods ConA Sepharose 4B affinity column was used to purificate mannoprotein from fermentation culture supernatant of CAP67.Results Up to milligram of mannoprotein was obtained from fermentation supernatants of Cryptococcus neoformans acapsular strain CAP67.Conclusion Fermentation method can be used to culture Cryptococcus neoformans, and the fermentation supernatants can be used for the purification of mannoprotein,which possessed the superiority of shorter time-consuming and higher yields compared to the traditional method.

Antifungal susceptibility of 30 clinical isolates of Rhodotorula spp. and literature review

JIANG Li-xiao, HUANG Qin, SHANG Pan-pan, SUN Ting-ting, LI Dong-ming

2012, 7(6): 342-347.

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Objective To test and analysis the antifungal activity of 5 drugs against clinical isolates of Rhodotorula spp. in vitro, and review the drug susceptibility data of another 249 strains of Rhodotorula spp..Methods Rhodotorula spp. were isolated and identified from clinical fungi bank of our hospital, susceptibility profile of these strains were tested with ATB Fungus3. A systematic MEDLINE search of English literature was performed to identify articles on Rhodotorula antifungal susceptibility in the past 20 years.Results Our results show that itraconazole, fluconazole and voriconazole are almost inactive against all of Rhodotorula spp.in vitro, Amphotericin B and flucytosine exhibit good antifungal activity,which is consistent with literature review.Conclusions The results show that, commonly used azole antifungal agents are inactive against Rhodotorula spp..Amphotericin B and Flucytosine are both active in vitro, being rational choice for empirical treatment.

Monitoring and analysis of pathogenic bacteria in 183 cases tinea capitis in Jiujiang and surrounding areas

SUN Xiao-hong, HONG Xia, SHUAI Li-hua, SONG Qiu-he, PAN Bin, HE Wei-wei

2012, 7(6): 348-351.

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Objective To understand tinea capitis in type and distribution of pathogen in Jiujiang and surrounding areas in three years.Methods Using retrospective methods to analysis 183 cases dermatopathic outpatients of tinea capitis in our hosipital from 2008 May to 2011 June.Results There were 183 cases with age from 5 months to 71 years, 164 cases (89.6%) were from 2 to 13 years old. There were tinea alba 138 (75.4%) cases; kerion 28 (15.3 %) cases; black-dot ringworm 17 (9.3%) case; 144 (78.7%) with postive microscopic and 154 (84.2%) with postive culture. The main pathogenic fungi were Microsporum gypseum 41 (26.6%),Trichophyton mentagrophytes 36 (23.4%),Trichophyton rubrum 33 (21.4%),Microsporum canis 24 (15.6%) and Trichophyton violaceum 12 (7.8%).Conclusions Children were the main tinea capitis patients in Jiujiang and its surrounding areas, tinea alba is the highest one, and Microsporum gypseum is the most common pathogenic fungi,Trichophyton mentagrophytes,Trichophyton rubrum and Microsporum canis contributes second, thrid and fourth respectively.

Analysis of etiology and clinical data of 106 onychomycosis cases in Shijiazhuang

ZHU Jing-xian, HAO Hong-yi, Li Li-cui, LIN Yuan-zhu, WAN Li, GAO Shun-qiang

2012, 7(6): 352-355.

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Objective To study the clinical classification and pathogenic fungi and to analyze the related factors with regard to the patients of onychomacosis in the Fourth Hospital of Hebei Medical University.Methods The patients were from dermatology outpatient within the period from January to December in 2011. In each case, the most serious nail was selected as target nail and the method of multiple point fungal culture was adopted. The fungal culture examination was positive and diagnosis was onychomycosis. The strains were identified to species. Onychomycosis-related factors were logged into the questionnaire.Results The clinical classification of all the 106 cases of onychomycosis were DLSO 47.17%, TDO 45.28%, PSO 4.95%, and SWO 2.97%. There were 111 fungal stains including dermatophytes 72 (64.86%), yeast 34 (30.63%), and NDM (nondermatophyte moulds, NDM)5 (4.50%). The patient's age (year) and duration (month) showed positive correlation: the older the age, the longer the duration. The number of infected nails and duration were positive correlation, that is, longer duration, more numbers. The most common nail changes were discoloration (84.91%) and thickening (74.53%). Single nail infection was found in 101 cases, accounting for 95.28%; mixed fungal infection was found in 5 cases that there were two kinds of fungi existing together in one patient, one of them was dermatophytes, and the other was yeast.Conclusion Onychomycosis was a chronic progression disease. The clinical classification of onychomycosis was complex and the pathogenic funguses were multiple. So the disease shall be paid much attention and given early treatment.

A case of Chromoblastomycosis caused by Fonsecaea pedrosoi and literature review

LIU Fang, FENG Jiao, KONG Qing-tao, WU Fan, REN Fang, SHEN Yong-nian, LV Gui-xia, SANG Hong

2012, 7(6): 356-358,364.

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Reported one case of left opisthenar and wrist chromoblastomycosis caused by Fonsecaea pedrosoi ,and statistical analysis the epidemiology and clinical characteristic in our country through literature retrieve. A 48-year-old man with infiltrated erythematous plaque,verrucous hyperplasia accompany with pruritus in left opisthenar and wrist for ten-years. Lesions Histopathology showed mixed inflammatory cell and multinucleated giant cells infiltrate with sclerotic bodies in the dermis. Positive periodic acid-schiff staining of sclerotic bodies. The isolate was identified as Fonsecaea pedrosoi by fungal examination, PCR and sequence analysis. The patient was treated with itraconazole 600mg daily for 3 months, with a complete resolution of the dermatosis.

A case of facial skin candidal granuloma

ZHU Wen-jing, LI Xue, ZHANG Xu-yan, MU Yan, JIN Xian-hua, YU Kai, LI Fu-qiu, XIA Jian-xin

2012, 7(6): 359-361.

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A case of facial skin candidal granuloma was reported.A 14-year-old girl had a history of erosion around mouth for 10 years and interspersed plaques,scab on face for six years.Physical examination showed dark-red plaques with tan thick scab of various sizes on her face. Fungal microscope examination showed mass hyphae and that was identified as Candidas albicans.Histopathological examination revealed infiltration of diffuse lymphocytes,sparse neutrophils and multinucleated giant cells in upper dermis.Mass hyphae was found in cuticle and spores were found in multinucleated giant cells by PAS stain.The diagnosis was made as candidal granuloma.The lesions improved after treatment of two-month with Griseofulvin,six-month with BCG-PSN,seven-month with Itraconazole, most of the scab was dropped. The patient is still under treatment.

A case of AIDS with erythroderma and Candida infection as the primary manifestations

YI Xue-mei, DING Yang-feng, TU Hong-gang

2012, 7(6): 362-364.

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A case of AIDS characterized by erythroderma was reported. A 64-year-old man had a history of whole body rash for two months and fever for a week. A severe body weight loss, conjunctival congestion, white dense guttate pseudomembranous of the palate and buccal mucosa, and diffuesed dark erythema with brown crusts scaling throughout the body were found in the examination. According to Oral Mucous Membrane smear being positive for fungi and HIV-1, the man was diagnosed as AIDS, erythroderma and oral candidiasis. In the end, he died of severe pulmonary infection.

Adjuvant management of patients with candida vulvovaginitis

CHANG Lu-lu, HU Su-quan, ZHU Hong-di, WU Guo-hua

2012, 7(6): 365-366.

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Objective To report 80 cases of candida vulvovaginitis received adjuvant management.Methods The adjuvant management is including methods of psychological counseling, guidance, medication and health education.Results Seventy-two cases (90.00%) recovered after a course of treatment, 6 cases (7.50%) were cured after two courses and 2 cases (2.50%) changed into the refractory candida vulvovaginitis.Conclusion Psychological counseling, guidance, medication, skin care and health education are benefit for the treatment of patients with candida vulvovaginitis.

Glucocorticoids and invasive fungal infections:a review

HAO Fei, WANG Li, ZHOU Cun-jian

2012, 7(6): 367-371.

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It is well-known that hyperglucocorticoids play an important role in inducing the fungal infection,especially opportunistic fungal infection,which is attributed to the complex dysregulation of immunity caused by glucocorticoids.The patients, both those with endogenous Cushing's syndrome and, those receiving exogenous glucocorticoid therapy, are especially at risk of invasive fungal infections. A better knowledge of the interplay between glucocorticoid-induced immunosuppression and invasive fungal infections should assist in earlier recognition and establishment of specific prevention and treatment strategies in order to improve outcomes of such infections.

Advances in gene knockout technology of Candida albicans

TAN Hong-yue, CHEN Li-hua, HUANG You-ming, ZHU Hong-mei, GU Jun, WEN Hai

2012, 7(6): 375-379.

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Nowadays,candidiasis has been one of the most common hospital-acquired infection,and Candida albicans is the most important pathogen.As C.albicans is capable of developing drug-resistance, understanding the overall mechanism of C.albicans pathogenesis, and further finding new drug targets are important goals. To study this pathogen, it is advantageous to manipulate its genome. This review focused on recent progress of C.albicans gene knockout strategies, and discussed their merits and drawbacks.

Advances of Candida albicans biofilm construction in vitro and in vivo

SHAO Jing, WANG Chang-zhong, WANG Tian-ming, CHENG Hui-juan

2012, 7(6): 380-384.

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The chronic and refractory candidal infections caused by Candida albicans have close relations with its biofilms formation. Many biological characteristics of biofilm are different from those of planktonic counterparts. Proper constructions of biofilm model are critical to the researches of the mechanism of biofilm formation and medicinal intervention. In this paper, the authors make a comprehensive review of in vitro and in vivo biofilm constructions of Candida albicans at home and abroad according to their studies.

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