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    28 December 2015, Volume 10 Issue 6
    论文
    The inhibitory effects of Butyl alcohol extract of BaiTouWeng decoction on biofilm formation of Candida albicans
    ZHANG Meng-xiang, LU Ke-qiao, XIA Dan, XIA Xue, SHI Gao-xiang, SHAO Jing, WU Da-qiang, WANG Tian-ming, WANG Chang-zhong
    2015, 10(6):  321-327. 
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    Objective To investigate the effects of Butyl alcohol extract of BaiTouWeng decoction (BAEB) on the biofilm formation of Candida albicans clinical strains isolated from vulvovaginal Candidiasis (VVC).Methods Microdilution methods was used to determine the MIC.XTT reduction assay was applied to determine the SMIC80.Time-kill curve method was applied to detect the effects of BAEB on living cells of Candida albicans.Crystal violet staining method was used to determine the biomass of the biofilm.Scanning electron microscopy (SEM) was applied to observe the morphological changes of the biofilm.Confocal laser scanning microscopy (CLSM) was applied to determine the thickness of the biofilm.The quantification real-time PCR (qRT-PCR) was used to detect expression changes of genes (HSP90,UME6 and PES1) of the biofilm treated by BAEB.Results The MICs of BAEB against C.albicans strains are determined as 64~256 μg/mL.The SMIC80s of BAEB against the biofilm of C.albicans strains are determined as ≥ 1024 μg/mL.Time-kill curve results indicate that BAEB has a promise antifungal effect at concentrations of 512 and 1 024 μg/mL.Crystal violet staining results show that the biomass of C.albicans is reduced by BAEB at 512 and 1 024 μg/mL.SEM results indicate that the formation of C.albicans biofilm carriers is inhibited by BAEB on different adhesion,and the morphology of biofilm is also affected by BAEB.The thickness of C.albicans biofilm is reduced by BAEB according to CLSM results.Furthermore,qRT-PCR results indicate that expression of UME6 is significantly down-regulated by BAEB at 256,512,1 024 μg/mL,and HSP90 is up-regulated at 512 and 1 024 μg/mL of BAEB,and PES1 is not affected by BAEB at any concentration.Conclusion BAEB inhibits effectively the biofilm formation of VVC strains of C.albicans.

    Effect of mandelic acid and its metabolism made on virulence of Cryptococcus neoformans
    ZHOU Zhao-jing, FANG Wei, PAN Wei-hua, LIAO Wan-qing
    2015, 10(6):  328-331. 
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    Objective To explore the effect mandelic acid and its metabolism made on virulence of C.neoformans.Methods Constructed recombinant fragments of mandelate racemase,mandelate dehydrogenase and hydrogen peroxide copper amine oxidase by nested PCR,and put them into JEC21 through gene gun.Identified the positive clones by PCR,DNA sequencing.What's more,detected mutant strains in vitro responses to various stress conditions.Results Successfully constructed mutant strains (mreΔ,lmdΔ,pcaoΔ),and found mreΔ took little effect on virulent factors,pcaoΔ and lmdΔ showed weaker in high temperature,high permeability,oxidative stress but secreted more melanin.Conclusion Mandelic acid and its metabolism play role in mediating the virulence of C.neoformans,but the exact mechanism needs further study.

    Identification and knocking-out of polarized growth factor gene kipA in Aspergillus fumigates
    ZHU Jun-hao, LI Li, ZHANG Qiang-qiang, CHEN Yu-chong
    2015, 10(6):  332-335. 
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    Objective To construct the kipA mutant strain.Methods The vector containing upper and downstream genes of kipA linked with hygromycin resistance gene in the middle was constrctied via In-Fusion assembly.The kipA gene of Aspergillus fumigatu was replaced with hygromycin resistance gene via the method of homologous recombination.The mutant strain was identified and validated by realtime-PCR.Results The kipA deletion strains were successfully constructed.Conclusions Successful construction of kipA mutant strain would provide the foundation for the research of polarized growth and vascular invasion of Aspergillus fumigatus.

    The evaluation of Aspergillus fumigatus infecting human bronchial epithelial cell in vitro
    SHEN Liang-liang, GAO Qing-yun, HUANG Xin, ZHENG Jian-feng, HAN Yu, ZHAO Jing-jun
    2015, 10(6):  336-339. 
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    Objective To establish a model for the relative research that simulate Aspergillus fumigatus invading human bronchial epithelial cell (HBE) and analyze the change of HBE in different time.Methods Using Aspergillus fumigatus to infect HBE in vitro and researching the change rules (biological form of HBE by adding liquid supernatant from the model,the apoptosis rate of the cells by Fluorescence Activating Cell Sorter and apoptosis gene (Bax) expression by real-time PCR).Results As the action time of Aspergillus fumigatus infecting HBE gone by,the biological form of HBE has changed,the retractile and round cells grew in number,the apoptosis rate of the cells also increased gradually and BaK expression obviously increased.Conclusion The changes of biological form,apoptosis rate and the Bax expression of HBE are evaluation system of simulating Aspergillus invading HBE in IPA which establishes in vitro and can use for relative basic and clinical research.

    Protective effect of thioredoxin reductase GliT of Aspergillus fumigatus in a murine model of invasive aspergillosis
    LIU Qian, SHI Li-ning, SHANG Xiu-juan, CHEN Xiao, HU Yu-an, CHEN Yong, LI Fang-qiu
    2015, 10(6):  340-344. 
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    Objective To investigate the immunoprotection in mice immunized with thioredoxin reductase GliT of Aspergillus fumigatus.Methods C57BL/6 mice were randomly assigned into two groups:immunization group intramuscularly immunized with recombinant TR for two times;control group intramuscularly immunized with PBS instead of TR.Survival rate,A.fumigatus lung burdens,pathologic examination and cell count of alveolar lavage were evaluated.Results The survival rate of immunization group was 64.7% within 7 days after infection while that of control group was 0.The medians of CFUs from the lung tissues of the suvivors and the nonsuvivors were 0 and 44 (P25,P75 were 21,70) respectively (P<0.01).Pulmonary pathology indicated that the lungs of the dead immunized mice and control mice showed alveolar hemorrhage,inflammatory cells recruitment,focal tissue necrosis and hyphal elements throughout the entire lung.In contrast,the lung tissues of the survivors were damaged slightly and free of hyphal elements.Alveolar lavage fluid smear showed that the counting of the survival is given priority to with mononuclear cells,accounting for about 84.2%,neutrophils accounted 15.8%.The counting of the nonsurvival is given priority to with neutrophils,accounting for about 66.4%,while 33.6% mononuclear cells.Conclusion Recombinant TR of Aspergillus fumigatus can induce a modest protection in a murine model of invasive aspergillosis and might be a potential protective antigen.

    A Study of fluconazole resistance mechanism of Candida albicans based on Hog-MAPK signal pathway
    WEN Yi-yang, HUANG Xin, SHEN Shuai-shuai, ZHAO Jing-jun
    2015, 10(6):  345-351. 
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    Obejectives To investigate the role of Hog-MAPK signaling pathway in the fluconazole resistance mechanism of Candida albicans mechanism.Methods Using Real-Time PCR and Western blot to compare the expression of HOG1 gene and Phosphorylation p38 MAPK protein from Hog-MAPK signaling pathway in fluconazole sensitive strain and resistance strain,then dectect the fluconazole MIC of Candida albicans HOG1 mutant and its original parent strain/standard strain.Results The difference of the expression of Hog-MAPK signaling pathway related gene and protein is found between Candida albicans fluconazole sensitive strain and resistant strain,the expression of sensitive strain is less than resistant strain to a certain extent,and HOG1 mutant strain is more sensitive to fluconazole.Conclusion The fluconazole resistance mechanism of Candida albicans may be associated with some gene and protein from Hog-MAPK pathway.The decreased expression of these genes and proteins may cause change of resistance to fluconazole.

    Five cases of nasal sinus mycoses clinical and laboratory study
    YAO Li-min, ZHANG Xue, WAN Zhe, WANG Quan-gui, YU Jin
    2015, 10(6):  352-356. 
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    Objective To report five cases of nasal sinus mycoses caused by filamentous fungi and related laboratory investigation.Methods Analyze five clinical cases of nasal sinusitis.Direct smear examination,fungal culture,PCR and sequence analysis were performed for diagnosis.Results Morphological study could not identify fungi to species level but sequences analysis can identify the 5 isolates as Fusarium solani,Aspergillus fumigatus,Aspergillus flavus,Pseudallescheria boydii,Fusariam napiforme.Endoscopic surgeries were applied to 4 patients;1 patient received voriconazole and amphotericin B therapy;treatments were effective to all patients.Conclusions Nasal sinus mycoses could be cause by various fungi.Molecular Biology can identify pathogens to species level,and endoscopic surgery is a reliable treatment for nasal sinus mycoses.

    Posaconazole for prophylaxis of invasive fungal infections:a meta-analysis
    HUANG Jin-Long, XIAO Shi-Ji, TIAN Li-Hong, ZHANG Yuan-Yuan, ZHOU Hua-Rong, SHEN Jian-Zhen
    2015, 10(6):  357-364. 
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    Objective To assess the efficacy and safety of posaconazole for prophylaxis of invasive fungal infections.Methods Databases such as PUBMED,Medline,Embase,Cochrane Library,SinoMed,CNKI,VIP Information and conference proceedings were searched for relevant randomized controlled trials (RCTs) about posaconazole for prophylaxis of invasive fungal infections.The cut-off time is December 2014.And reference of the included articles were also manually searched.Quality of included studies were assessed and data of which were extracted.Both Review Manager 5.2 and Stata 12.0 software were used.Results Four RCTs which included 1 476 patients met the search criteria were assessed.Meta analysis results:the IFI rate[RR=0.41,95%CI (0.27,0.63),P<0.000 01],especially in prophylaxis for invasive aspergillosis (posaconazole group significantly reduced:the IFI rate,especially in prophylaxis for invasive aspergillosis (posaconazole vs fluconazole,P<0.000 1);and mortality from IFI.However,no significant differences were observed in adverse reactions,therapy discontinued because of adverse reactions,all-cause mortality rate between posaconazole and controlled groups (P>0.05).For subgroup analysis,there was no significant difference in both the rates of adverse reactions and invasive candidiasis infection between posaconazole and fluconazole (P>0.05).Conclusion Posaconazole can be used as a good alternative of amphotericin B lipid complex,fluconazole or itraconazole for prophylaxis of invasive fungal infections.Posaconazole significantly improves the effect of prophylaxis and decreases IFI associated mortality but with no risk of increase in adverse events.

    A case of generalized superficial mycosis with SLE
    GUO Yan-yang, WU Zhuo-yan, GAO Ji-xin, ZHAO Di, LIAO Wen-jun, WANG Sheng-chun
    2015, 10(6):  368-370. 
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    A 57 years old woman with 18 years long systemic lupus erythema history had developed erythema and scales with itching for 3 years.Fungus microscope examination disclosed mycelium and fungus culture indicated the Trichophyton rubrum.The patient was treated by oral itraconazole capsules and topical Sertaconazole Nitrate Cream to tinea corporis,and topical Sertaconazole Nitrate Cream following CO2 fractional laser treating to onychomycosis.The lesions resolved with negative fungal check after continuous treatment for 9 weeks to tinea corporis and 9 weeks to onychomycosis,respectively.

    Progress of fungi in gut and the related diseases
    NIU Hai-jing, SU Bing-zhong
    2015, 10(6):  377-379,384. 
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    There were 66 fungal genera and 184 fungal species (with Candida as the dominant fungal genera) colonizing in human gastrointestinal tract.Similar to commensal bacteria,commensal fungi could be an important part of microbiome,which helps organ developing,participates of nutrients metabolism,induces immunity and triggers (or represses) inflammatory responses.When specific defense systems damaged (e.g.immune deficiency) or microbiome unbalanced (e.g.after administration of antibiotics),opportunistic pathogenic fungi could contribute to mycotic esophagitis in susceptible individuals,and might also play a role in gastrointestinal diseases such as infective colitis,irritable bowel syndrome,inflammatory bowel disease,and acute pancreatitis.

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