Table of Content

28 April 2012, Volume 7 Issue 2

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Construction and identification of recombinant adenovirus of mannose receptor gene

LI Ping, WEN Hai, ZHU Hong-mei, XU Hong, HU Chan, TAN Hong-yue

2012, (2):  70-72. 

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Objective To clone and construct recombinant adenovirus of mannose receptor(MR) gene from RAW264.7 macrophage cell line.Methods PCR technique and recombination methods were used in cloning and construction recombinant adenovirus of MR gene, Western blot was used to verify the MR protein expressed in HEK293.Results MR gene was amplified from genomic DNA of macrophage and cloned into pShuttle-CMV vector.The linearized pShuttle-CMW plasmid was cotransformed with the pAdEasy-1 vector into the BJ5183 cells,producing the recombination adenovirus which can express the MR protein.Conclusion Successful cloning of MR gene and construction of MR gene expression recombinant adenovirus will be useful for the further research on the functions of MR in Dendritic cells.

In vitro activity of terbinafine in combinations with other antifungi drugs against clinical pathogenic fungi of vulvovaginal candidiasis

JI Ya-juan, WANG Ai-ping, LIU Wei, WAN Zhe, LI Ruo-yu

2012, (2):  73-76,81. 

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Objective To investigate the in vitro activity of terbinafine(TEB)in combinations with, itraconazole(ITC) fluconazole(FLC), clotrimazole(CMZ); miconazole(MCZ) and nystatin(NYS) against clinical isolates of vulvovaginal candidiasis.Methods The minimal inhibitory concentrations(MICs)of TEB, FLU, ITR, MCZ, CMZ and NYS against 46 strains of Candida were determined by using CLSI broth microdilution method(M27-A3).Additionally, the interactions of TEB with ITR,FLU,MCZ, CMZ and NYS, were assessed by using the checkerboard technique. The fractional inhibitory concentration index(FICI) was used to categorize drug interactions as following, synergy, FICI≤0.5; indifference, FICI>0.5 and≤4.0; or antagonism, FICI>4.0.Results The MIC ranges of TEB, FLU, ITR, MCZ, CMZ and NYS against candida. spp were 0.5-256 μg/mL,0.5-128 μg/mL,0.5-32.0 μg/mL,0.125-16 μg/mL,0.03-4.0 μg/mL and 2-8 μg/mL, respectively. The combinations of TEB with FLU, ITR, MCZ, CMZ showed synergic effects are 20/20(100%), 16/20(80%), 11/20(55%)and 18/20(90%)in C.albicans isolates, respectively. Besides, the combinations of TEB with FLU, ITR, MCZ and CMZ showed indifference against Candida krusei,Candida parapsilosis and Candida glabrata. The combinations of TEB with all the azoles showed synergic for Candida guilliermondii. The combinations of TEB with FLU, ITR, MCZ, CMZ showed synergic effects are 67%,60%,20% and 20% in Candida tropicalis, respectively. There was not synergic effect when TEB combined with NYS in all strains.Conclusions Different Candida spp show different effects to the combinations of TEB with azoles. There is not synergic effect when TEB in combination with NYS for all strains.

Detecting CPH1,EFG1 gene expression of planktonic cells and biofilm of Candida albicans with fluorescent quantitative PCR assay

YANG Jun, ZHANG Tian-tuo, ZHU Jia-xin

2012, (2):  77-81. 

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Objective To investigate the role of gene expression of transcription factor CPH1 and EFG1 in biofilm formation by detecting the difference of CPH1,EFG1 mRNA expression between biofilm and planktonic cells of C.albicans.Methods Ten C.albicans strains isolated from respiratory tract were used. The total RNA of planktonic and biofilm cells were extracted separately, the mRNA expression of CPH1,EFG1 were measured with fluorescent quantitative PCR assay.The △△Ct that show relative value of mRNA expression were calculated.Results In this study,the mRNA expression of EFG1 in biofilm cells was 1.42~7.14 times higher than that in planktonic cells(P<0.05).The mRNA expression of CPH1 was up-regulated in biofilm cells of 8 isolates, down-regulated in 1 isolate and show no difference in 1 isolate(P>0.05).Conclusion The study show that transcription factors CPH1,EFG1 participated in the regulation of biofilm formation in clinical isolates, and further research in vivo is needed to provide more proffs.

Applications of PCR fingerprinting in the study of epidemiology of Candidal vulvovaginitis and balanitis

JIANG Ai-ping, HUANG Jian-qing, LU Wan-ding, XU Ming, WU Jian-hua

2012, (2):  82-84,115. 

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Objective To establish a molecular biological method of identifying infection, recrudescence or reinfection of candidal vulvovaginitis and balanitis.Methods PCR was used to amplify the microsatellite region of the two pathogens from patients infected twice or couples infected simultaneously. The products were analyzed according to PCR fingerprinting polymorphism to determine the relationship between the two strains.Results Of the 22 tested strains, 8 strains from 4 individual patients got a similarity module above 0.8, which was considered recrudescence; 6 strains from 3 pairs of couple also displayed a similarity module above 0.8, which was regarded as a result of sexual transmission. The rest got a similarity module below 0.8, which could be considered reinfection.Conclusion To some extent, the method of microsatellite primed PCR fingerprint spectrum could identify infection, recrudescence or reinfection, the result of which could be suggestive in clinical treatment.

Effects of TLR2 on IL-8 and γ-IFN secretion of keratinocytes in Trichophyton rubrum infection

TU Sheng-an, ZHANG Yong, CHEN Hui, CHEN Xing-ping

2012, (2):  85-88. 

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Objective Observe the change of γ-IFN and IL-8 secreted by keratinocytes after Trichophyton rubrum infection, and the effects of TLR2 on γ-IFN and IL-8 secretion. To explore the role of TLR2 in anti-dermatophytic infection.Methods Keratinocytes treated before and after anti-TLR2 monoantibody were stimulated by Trichophyton rubrum.The levels of γ-IFN and IL-8 in supernatant were determined with ELISA at different time points.Results The levels of γ-IFN and IL-8 secreted by kerainocytes were significantly increased after induced with Trichophyton rubrum(P<0.05).The level of γ-IFN reached(85.36±4.54) pg/mL at the point of 4h,and(445.58±13.99) pg/mL at 16 h; After treated with anti-TLR2 monoantibody, the levels of IL-8 were reduced at the points of 2 h,4 h,8 h and 16 h(P<0.05); and the levels of γ-IFN were reduced at the points of 2 h, 4 h and 8 h(P<0.05), but had no statistically significant reduction at the point of 16 h(P>0.05).Conclusion After stimulated keratinocytes,Trichophyton rubrum can promote its secretion of γ-IFN and IL-8; TLR2 play an important role onγ-IFN and IL-8 secretion of keratinocytes.

Illegible sporotrichosis on the face

XU Yang, LI Fu-qiu

2012, (2):  89-91. 

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To explore the nonspecific clinical manifestations of sporotrichosis. Sporotrichosis is a deep fungal disease with common cutaneous manifestations as chronic granuloma, ulcer and nodular lesions. Part of patients may not show the specific clinical manifestations becarse of miscellaneous reasons. This situation above increase the difficulty of final diagnosis to some dermatologist with little related clinical experience. The eight patients of sporotrichosis to be discussed in this paper didn't have the same clinical manifestation as those mentioned characteristic in a previous articles.This situation suggestes that the definite diagnosis should be closely according to the medical history, the living environment, and even the process of diagnosis and treatment.

Three cases of human tinea corporis transmitted from rabbits caused by Trichophyton mentagrophytes

KONG Qing-tao, SANG Hong, ZHANG Min, LIU Fang, WANG Xue-lian, WANG Gao-feng

2012, (2):  95-97. 

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Report three cases of human tinea corporis transmitted from rabbits caused by Trichophyton mentagrophytes. Three patients all had pruritic annular erythema with squam and a history of close contact with pet rabbits. Direct microscopic examination of scales revealed hyphae. Fungal cultural lead to the identification of T.mentagrophytes. They were all cured by oral and(or) topical antifungal therapy.

A case of fixed cutaneous sporotrichosis presented with hyperplasia and crusts

LI Yun, MAO Xiao-fei, DU Wei, LIU Yue-hua, SUN Qiu-ning

2012, (2):  98-100. 

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Sporotrichosis is a chronic subacute infection caused by fungi belonging to the Sporothrix Complex.Sporothrix schenckii,as the causative agent, can be isolated from soil, wood, land, and even marine animals. We report a case of cat-associated sporotrichosis. A 16-year-old boy with fixed cutaneous spomtrichosis caused by Sporothrix schenckii presented with hyperplasia, exudation and crusts. We present the clinical manifestation, histopathological presentations, microscopic examination and fungal culture of sporotrichosis. After seven monthes treatment of potassium iodide and terbinafine ointment,the lesion disappeared.

Atypical favus in child:a case report and literature review

LI Li, ZHENG Dong-yan, WANG Yun-ru, ZHU Jun-hao, ZHANG Qiang-qing

2012, (2):  101-103. 

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A case report of atypical favus in child caused by Trichophyton schoenleinii.With mild clinical symptoms, the child lacked some typical clinical signs as: concave disc formed sulfur-yellow crusts, characteristics of the rat smell,et al. Direct microscopic examination of the infected hairs found a large number of irregular hyphae, intercalary swollen spores with unequal size and irregular shape Trichophyton schoenleinii was growth on fungal culture media and confirmed by DNA sequencing. Drug susceptibility(E-test) results showed the isolated strain was sensitive to itraconazole, amphotericin-B, caspofungin and voriconazole. The lesions were recovered in two months by oral itraconazole capsule and topical 2% sertaconazole cream.

Clinical observation of sertaconazole nitrate cream in treatment of skin candidiasis in children

2012, (2):  106-108. 

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Objective To evaluate clinical efficacy and safety of sertaconazole nitrate cream in the treatment of children's skin candidiasis(moniliasis).Methods One hundred patients were enrolled in the test and were divided into 2 groups. Sertaconazole nitrate cream was applied topically to the affected areas of the body twice daily for 14 days in the treatment group.Miconazole nitrate ointment was applied topically to the affected areas of the body twice daily for 14 days in the control group.The efficacy and the adverse reaction were observed.Result Treatment group's curative rate is 92.00% and control group's 64.00%(P<0.01).Compared with the control group, the treatment group produced the effect more quickly with a shorter cured period(P<0.01).There was little adverse effect observed in the test.Conclusion Sertaconazole nitrate cream acts effectively and safely on children's skin candidiasis. It is worthy of application.

Recent insights from Dectin-1-Pattern Recognition Receptor

YI Xiu-li, LIU Bin, QI Xiao-long

2012, (2):  121-124. 

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The Dectin-1 is a non-Toll-like pattern recognition receptor expressed predominantly by myeloid cells, which can induce its own intracellular signalling and can mediate a variety of cellular responses, plays a key role in anti-fungal immunity. The recent studies have focused on the signalling pathways from Dectin-1, interactions between Dectin-1 and other PRRs, Dectin-1 and adaptive immunity and the role of Dectin-1 in anti-microbial immunity.

Biofilm gene expression and biofilm-related genes of Candida albicans:an update

SHANG Qing-hua, CAO Ying-ying, MIAO Hao, JIANG Yuan-ying

2012, (2):  125-128. 

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Candida albicans is an opportunistic fungal pathogen with the ability to form biofilms on in-planted medical devices. Cells in biofilms display a phenotype that is markedly different from their planktonic and free-living counterparts in both form and function.Recent advances in microarray and genetic manipulation have begun to clarify the mechanisms that govern C.albicans biofilm development and acquisition of such unique phenotype. These studies are considered to be important in finding new targets of antifungal drugs.