Table of Content

28 October 2013, Volume 8 Issue 5

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The effect of prototheca on the differentiation of mice Th17 cells

KANG Yu-li, ZHAO Ying, ZHANG Qiang-qiang

2013, 8(5):  257-260. 

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Objective To study if prototheca could influence the differentiation of mice Th17 cells and the secretion of Th17 associated cytokines.Methods Prototheca was cultured in vitro, and T lymphocytes were isolated from spleen of mice by nylon wool column, and cultured in vitro. Prototheca and T cells were co-cultured in transwell plate with a ratio of 1:5. Cells were culture for 2 h,4 h,8 h,12 h,24 h,48 h,and then T cells and supernatant were collected respectively, and the ratio of Th17 cell was analyzed with flow cytometry, and the level of IL-17 in the supernatant was detected with ELISA.Results From 2 h on, with the prolonged incubation time, the proportion of Th17 cells decreased gradually, and it became stable 8 h later (P<0.05); the level of IL-17 in the supernatant increased at earlier 2-8 h, and decreased from 8 h on, and it became stable 24 h later (P<0.05).Conclusion Being co-cultured with T cells, Prototheca could inhibit the differentiation of mice Th17 cells, but it could promote the secretion of IL-17 at the earlier 2-8 h.

The effect of Cryptococcus neoformans capsular GXM on energy metabolism and apoptosis of mouse microglia

ZHOU Jie, PAN Wei-hua, LIAO Wan-qing

2013, 8(5):  261-264. 

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Objective To detect the effect of Cryptococcus neoformans GXM on energy metabolism and apoptosis of mouse microglia cell.Method ①The capacity of ATP generation in each group was evaluated with UV spectrophotometric determination.②Annexin V-fluorescein isothiocyanate/propidium iodide double label Method was taken to detect apoptosis rate of BV-2 cells after co-incubation with GXM.Results ①The capacity of ATP generation decreased after intervention by GXM.②GXM was able to induce apoptosis in microglial cells in vitro.Conclusion Cryptococcus neoformans could exert effect on host cells via induction of energy metabolism disorder and apoptosis in microglia by GXM.

The study of a simple in vitro onychomycosis model

HU Chan, ZHU Hong-mei, TAN Hong-yue, LI Ping, ZHAO Jin, CHEN Li, WEN Hai, SONG Wei-Fang

2013, 8(5):  265-268. 

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Objective To determine a simple, novel and economic in vitro onychomycosis model.Method An innovatively designed "O" ring was applied to localize the fugal suspension on the ventral surface of the porcine hoof, then 50 μL of fungal suspension was applied. The hoof membranes were cultured in sterile petri dishes. The dynamic penetration of hoof plates was observed with gross appreance and histopathological examination.Result In this study we developed a novel in vitro model of onychomycosis with porcine hoof in which we observed the Trichophyton rubrum,Trichophyton mentagrophytes and Candida albicans dynamic permeation to the dorsal of porcine hoof both by gross appreance and histopathological examination and this model also enables one to evaluate how the fungi behave in the nail plate and changes in the localization.Conclusion The model facilitates monitoring of the nail pathogenicity.

Efficacy evaluation of 5% amorolfine nail lacquer topical treatment in the rabbits’ onychomycosis model

YANG Lian-juan, MO Xiao-hui, YANG Hong, GAO Zhi-qin, ZHAGN Chu-guang, YU Qian, ZHU Hong-mei, WEN Hai

2013, 8(5):  269-273. 

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Objective To establish an animal model of onychomycosis and evaluate the efficacy of 5% amorolfine nail lacquer in the model.Methods We applied Trichophyton mentagrophytes to the nails of the hind limbs of rabbits to establish an experimental,in vivo model of onychomycosis. The nails were taken from the rabbits' feet to histopathological examination after infection success. The growth form of the fungi was evaluated histopathologically. Then 5% amorolfine nail lacquer was applied to the nail for 4 weeks and the efficacy was evaluated by the fungal semiquantitative culture method.Result We successed to establish the onychomycosis model of rabbits that underwent steroid treatment. A cloudy white or pallide-flavens appearance like that of human onychomycosis was observed in the infected nails after inoculation one week. On histopathological examination, hyphae of T.mentagrophytes were seen to penetrate the nail plate, reached the nail bed. The total infection rate at 2, 4, and 6 weeks after inoculation was 52.78%,77.78% and 83.33%, respectively. The culture-positive rate at 2 and 4 weeks after topical treatment was 22.22% and 66.67%, respectively.Conclusion We successed to establish an onychomycosis model in using rabbits and evaluated the efficacy of a topical antifungal agent in this model. The efficacy of 5% amorolfine nail lacque in this model was 66.67% after 4 weeks.

Evaluation of Rapid ID Yeast Plus and API20C AUX systems in identification of yeast species,using molecular identification as ‘gold-standard’

FAN Xin, WANG He, SU Xiao-xiang, XIAO Meng, MA Xue-fei, MA Chao-yue, LIU Xu-guang, MA Ran, WU Yue, CHEN Yan-zhao, XU Ying-chun

2013, 8(5):  274-280. 

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Objective To conduct performance assessment on two commercialised kits for yeast identification: Rapid ID Yeast Plus (RapID YS) and API20C AUX (API20C), using molecular Methods as "gold standard".Methods A total of 194 strains of 2 yeast species were selected from China Hospital Invasive Fungal Surveillance Net (CHIF-NE) 2010 strain collection, which includes 130 strains of the five commonest species (Candida albicans,Candida tropicalis,Candida glabrata,Candida parapsilosis,Cryptococcus neoformans) that constitute 67% of the total. All strains were identified to species level by molecular methods. Identification by RapID YS and API20C were parallel conducted according to the product manufacturer's instructions.Results Amongst the isolates studied, 181 (18 species) were within the RapID YS identification database, and 87.8% (19/181 isolates) was correctly identified to species or species complex level. Comparatively, API20C database compassing a total of 174 strains (18 species), and 92.0% (160/174 isolates) was correctly identified to species or species complex level. For the five commonest yeast species, the correct identification rate of RapID YS and API20C, to species level, was 93.1% and 97.1%, respectively. For spe-cies not included in identification database, the incorrect identification rate was 23.1% (3/13) for RapID YS, and 60.0% (12/20) for API20C AUX. Overall, no significant difference was observed between two commercial Methods (McNemar test,P>0.0).Conclusions he identification capability of two commercial Methods were basically the same. RapID YS held relatively greater advantages in terms of operational convenience and turn-around time for. It possesses comprehensive application prospects for clinical applications.

Evaluation of BD Phoenix^TM in identification of the yeast by using clinical yeasts

DOU Hong-tao, XIAO Meng, FAN Xin, YANG Qi-wen, WANG Yao, WANG He, LIU Juan, XIE Xiu-li, XU Ying-chun

2013, 8(5):  281-284. 

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Objective To evaluate the ability of BD Phoenix^TM in identification of the yeast isolated from clinical speciemen.Methods 18 strains of Candida albicans,22 strains of C.tropicalis,19 strains of C.glabrata,8 strains of C.krusei,20 strains of C.parapsilosis,4 strains of C.guilliermondii,1 strain of C.inconspicua,C.lusitaniae,C.norvegensis,C.catenulata,C.kefyr,C.haemulonii,C.rugosa respectively,3 strains of C.lipolytica,3 strains of C.pelliculosa,2 strains of Geotrichum capitatum,14 strains of Cryptococcus neoformans were tested. The total of 120 stains were all isolated from clinical specimen. They were identified by using automated Phoenix^TM 100 to test the BD Phoenix^TM. The fungus-specific universal primers ITS1 and ITS4 were used to amplify the rDNA of isolates mentioned above. The products of PCR were sequenced and analyzed, which were compared with BD Phoenix^TM as golden standard. MALDI-TOF MS were also used to identify all the tested strains.Results By using BD Phoenix^TM,1 strain of C.norvegensis,C.inconspicua,C.lipolytica,C.kefyr,C.rugosa can't be identified and 1 strain of C.catenulate,1 strain of C.krusei,2 strains of C.pelliculosa and 1 strain of C.lipolytica had the wrong results. The other results were correct. The precision of identification for BD Phoenix^TM was 92.5%. All the results were obtained in 17 hours. It took us less than 6 hours to identify C.albicans,C. tropicalis and C.parapsilosis without influence of proposed media. There was no difference between identification results of rDNA ITS sequencing and MALDI-TOF MS.Conclusions Most yeasts can be identified to the species level quickly and correctly by BD Phoenix^TM. But its ability of identifying some uncommon yeasts need to be tested.

Maintenance White Phoenix bolus therapy for recurrent vulvovaginal Candidiasis

GE Tan, LIU Yan, TENG Zong-rong, SHEN Jian

2013, 8(5):  285-288,293. 

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Objective White Phoenix bolus was used to explore and investigate a new optional management of recurrent vulvovaginal candidiasis (RVVC).Methods After inducing clinical remission and microbiological cure with clotrimazole suppositories given in two 500 mg doses at 72 hour intervals, 486 women with recurrent vulvovaginal candidiasis were randomly divided into three groups. After menstruation, group A were treated with intravaginal clotrimazole (500 mg) monthly for three months. Group B were treated with White Phoenix bolus, one pill daily for three monthly except menstrual period. The others were designated as Control Group. All women were followed by six months of observation without therapy. The outcome measure was the proportion of women in clinical remission at the end of the maintenance treatment and six-month follow-up period.Result The proportions of women who suffered with recurrence at the end of the treatment and 3,6 months follow-up in White Phoenix bolus group were 13.61%, 26.62% and 56.83%, similar to those in clotrimazole group (10.12%, 21.83% and 51.41%,P>0.05), and significantly decreased as compared with 40.27%, 57.30% and 68.54%, respectively, in the control group (P<0.001). The adverse effect was slight and could rarely be seen.Conclusion Long-term treatment with White Phoenix bolus can inhibit the growth of candida, boost up the immunity of host, control the local inflammation and reduce the rate of recurrence of symptomatic vulvovaginal candidiasis. The study may represent a safe and effective alternative to RVVC.

Research progress of host’s immune mechanism to resist Aspergillus infection

YAN Yu-yan, ZHAO Zuo-tao, LIU Hong-gang

2013, 8(5):  302-306. 

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With the wide use of broad-spectrum antibiotics, corticosteroids and immunosuppressive drugs, and the rising incidence of AIDS, diabetes etc, the number of immunocompromised patient is increasing. Follow this, invasive Aspergillus infections are increasing gradually and more attentive than before. To clarify the pathogenesis and find the effective treatment of Aspergillus infections are the common propositions for basic researches and clinical workers. Host's immunity plays an important role in the pathogenesis of Aspergillus infection.To master host's immune mechanisms of Aspergillus is the foundation of studying Aspergillus infection. In our article, progresses on the study of host's immune mechanisms against Aspergillus infections are reviewed.

Virulence factors and the host immune responses of Sporothrix schenckii

WAN Xue, HE Dan, WANG Li

2013, 8(5):  307-310. 

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Sporothrix schenckii is an important dimorphic fungi and it can cause Sporotrichosis which is a common invasive skin infection. A further understanding for the characterization of virulence factors and the host immune responses of this fungus should be done. It is very importantand significant for the study on their pathogenicity,prevention and treatment of this disease. Here we review briefly the advancement on the characterization of virulence factors and the host immune responses of Sporothrix schenckii.

New progress on the drug resistance mechanisms of Candida albicans

CAO Ying-ying, CAO Ying-ying, JIANG Yuan-ying

2013, 8(5):  311-315. 

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In recent years, more and more research about deep fungal infections has been reported, it has increasingly become the common complications during the clinical treatment of several important diseases, among which, the incidence of candidiasis remains unacceptably high. Although there are many kinds of antifungal drugs used in clinic, the drug resistance is becoming more and more serious, which has brought great challenge to the clinical treatment. Recent studies on the mechanism of drug resistance of Candida albicans showed new progress. In this article, the new mechanisms of drug resistance about C.albicans are reviewed briefly.