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    28 April 2014, Volume 9 Issue 2
    论文
    Correlation study on macrophage-derived cytokines and disease course in Cryptococcus neoformans infection mouse model
    LEI Wen-zhi, SANG Jun-jun, PAN Wei-hua, LIAO Wan-qing
    2014, 9(2):  65-69. 
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    Objective We detected the expression of macrophage-derived cytokines in macrophage of Cryptococcus neoformans infection mouse model to explore the role of those cytokines in pathogenesis of infection mouse.Methods A mouse model of Cryptococcus neoformans infection was established by inhalation of Cryptococcus neoformans through one nostril. On day 1, 4, 7, 11, 14, 18, 21 after inoculation, the infected lung were observed by histopathological analysis. Macrophage-derived cytokines (IL-6、IL-8、TGF-β、TNF-α)were detected by RT-PCR.Results Histopathological examination showed few Cryptococcus neoformans distributed in lung on 4 days and granuloma formation on 7 days after infection. There were significant inflammatory cellular infiltration in the lung tissue on 11 days and a large number of Cryptococcus neoformans in granuloma on 14 days after infection. At 18 and 21 days,diffuse distribution of Cryptococcus neoformans in the lung and the lung necrosis were observed. The mRNA expression of TGF-βand IL-6 reached to the peak at 14 days, then decreased gradually.Conclusion TGF-βwas involved in regulating the anti-fungal immunity in Cryptococcus neoformans infection mouse.

    Synergistic effect of baicalein in combination with fluconazole on Candida albicans biofilm
    ZHAO Liu-ya, JIANG Jing-chen, YAO Xiang-wen, CAO Ying-ying, JIANG Yuan-ying
    2014, 9(2):  70-74. 
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    Objective To investigate the effect of baicalein (BE) in combination with fluconazole (FLC) on Candida albicans biofilm formation.Methods The inhibition of C.albicans biofilm by BE in combination with FLC was determined by confocal scanning laser microscopy and XTT methods. The water-hydrocarbon two-phase assay was used to measure the cell surface hydrophobicity (CSH). The expression of CSH1, EFG1, HWP1, ALS1 mRNA was measured by Real time RT-PCR.Results BE in combination with FLC could inhibit the formation of C.albicans biofilm. The cell surface hydrophobicity in the BE and FLC-treated group was lower than that in the groups treated by BE or FLC alone. Consistent with this, BE and FLC-treated biofilm cells expressed lower levels of CSH1, EFG1, HWP1 mRNA than the cells grown in the presence of BE or FLC alone.Conclusions BE in combination with FLC could inhibit the formation of C.albicans biofilm.

    Antifungal susceptibility testing of clinical isolates of fluconazole-resistant Candida spp. and itraconazole-resistant Aspergillus funmigates to posaconazole
    WANG Xue-jie, WAN Zhe, LI Ruo-yu, LIU Wei
    2014, 9(2):  75-78. 
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    Objective To determine the in vitro susceptibility of fluconazole (FLU)-resistant Candida spp. and itraconazole (ITC)-resistant Aspergillus spp. to posaconazole (POS).Methods The MICs of FLU, ITC, voriconazole (VRC) and POS were determined according to CLSI M27-A3 and M38-A2 methods in eleven isolates of Candida spp. and three isolates of Aspergillus spp.Results MICs of FLU, ITC, VRC and POS against FLU-resistant Candida spp. were 64 μg/mL, 0.5~16 μg/mL, 0.25~2 μg/mL and 0.125~1 μg/mL. And MICs of ITC, VRC and POS against ITC-resistant Aspergillus spp. were >16 μg/mL, 1~2 μg/mL and 0.25~0.5 μg/mL.Conclusion POS is effective to FLU-resistant Candida spp. and ITC-resistant Aspergillus spp. in vitro.

    Expression and significance of Th9 and Th17 in the peripheral blood of patients with cryptococcal meningitis
    LEI Wen-zhi, SANG Jun-jun, PAN Wei-hua, LIAO Wan-qing
    2014, 9(2):  79-82,95. 
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    Objective To explore the role of Th9 and Th17 cells in the pathogenesis of CM by mesureing the ratio of Th9 and Th17 cells in CD4+T lymphocyte of patients with cryptococcal meningitis (CM).Methods Twelve CM patients without antifungal treatment and 12 healthy controls were selected. The peripheral blood was collected from patients prior treatment and three weeks after treatment and the healthy controls. The ratio of Th9 and Th17 cells in peripheral blood were analyzed by flow cytometry.Result The ratio of Th17 cells in patients with CM before treatment was less than those of healthy people, and patients who received treatment showed the increased level of Th17 cells which was higher than those in patients before treatment and the healthy controls. The ratioof Th9 also increased in the patients who received treatment while there was no significant different between the patients before treatment and the healthy controls.Conclusion Th17 immune pathway is an important immune mechanism against Cryptococcus infection in patients with CM. The lack of Th17 cells may play a role in the pathogenesis and treatment-resistant of CM.

    Malassezia species constitution in youth from some countries of Southeast Asia
    ZHANG Hao, XIONG Xin-cai, RAN Yu-ping, LIU Ting, CHEN Yu-juan
    2014, 9(2):  83-87. 
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    Objective To investgate the distribution of Malassezia microflora in youth from some countries of Southeast Asia.Methods Two hundred and eighty-five volunteers came from China, Nepal, India and Pakistan were included, which were healthy volunteers or patients related with Malassezia species. The samples were collected from normal skin of face or lesions skin and cultured on Leeming & Notman mediums. The isolates were identified to species through physiological and morphological characteristics.Results Eight species were identified out from 501 isolated strains, mainly included 203 strains (40.5%) of M.globosa, 95 strains (19.0%) of M.sympodialis, 80 strains (16.0%) of M.furfur, 55 strains (11.0%) of M.restricta and 31 strains (6.2%) of M.slooffiae. There were no obvious geographical differences between these four countries.Conclusions The most frequently isolated species were M.globosa in youth from some countries of Southeast Asia, which were healthy volunteers or patients related with Malassezia species.

    Post-antifungal effect of Meifute antifungal solution against fungal induced tinea pedis in vitro
    SHI Dong-mei, GU Ke-bing, SHEN Yong-nian, LÜ Gui-xia, LIU Wei-da
    2014, 9(2):  88-92. 
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    Objective To investigate antifugal activity and the post-antifungal effect (PAFE) of Meifute antifungal solution against fungal induced tinea pedis.Methods We detected the minimum inhibitory concentration (MIC) of Meifute antifungal solution against fungal common induced tinea pedis using the United States CLSI M27-A3 and M38-2 scheme. Time-killing curve and post antifungal effects of Meifute antifungal solution against Candida albicans (ATCC90028) as the indicator fungal were measured.Result The MIC range, MIC50 and MIC90 of Meifute antifungal solution against 57 species were 1:40~1:160, 1:80 and 1:40 respectively. The ranges of the solution against Candida albicans and dermatophytes were 1:40~1:80 and 1:40~1:160. This antifungal solution has a strong antifungal effect. The degree and speed of the killing fungal were changed with the decrease of drug concentration. The PAFE of 0.5MIC, 1MIC and 2MIC were 0.85 h, 2.1 h and 3.59 h respectively, and the prolonged of PAFE was positively correlated to the antifungal solution concentration.Conclusion Meifute antifungal solution has a fast, sustained effect against Candida albicans and dermatophtes. Comparing with the effect on Candida albicans, the anigfungal solution have stronger antifungal effect on dermatophtes. The antifunal solution also have a long period of PAFE. These results showed that Meifute antifungal solution can be widely used to treat the superficial fungal infection of dermatophytes and Candida albicans.

    Penicillium marneffei sepsis in an AIDS patient
    HUANG Qin, LI Dong-ming, SUN Ting-ting, SHANG Pan-pan, GE Jie, FU You-gang
    2014, 9(2):  99-101. 
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    Penicillium marneffei (PM) infection is prevalent in South-East Asia and Southern China, which has been considered an acquired immunodeficiency syndrome (AIDS)-defining disease. In this paper, we describe Penicillium marneffei sepsis in an HIV-infected patient without skin presentation. A 39-year-old man from Guangdong province came to our clinic with 1-month history of fevers and lymphadenectasis. He was found continuously decrease of leukocyte, erythrocyte and thrombocyte. Anti-HIV antibody screening test and confirmation test both appeared positive. Blood cultures were positive for fungi. Bone marrow biopsy showed intracellular septate spores. The P.marneffei was identified by rDNA PCR. The patient was diagnosed with AIDS and Penicillium marneffei sepsis. He was treated immediately with fluconazole and transferred to the infectious disease hospital. During the follow-up, he died of multiple organ failure. This case suggests that fungemia could be the initial symptom of penicilliosis marneffei, which should be paid enough attention by clinicians. Patient's blood should be strictly isolated because of its infectivity, and detection of HIV antibody is necessary. The patients with fever of unknown origin and unsafe sex history may be diagnosed as infection of HIV and Penicillium marneffei.

    Three cases of fungal keratitis caused by Candida
    XU Xue, HUANG Yun-li, ZHANG Dao-jun, LI Hong-bin
    2014, 9(2):  105-106. 
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    Fungal keratitis is a serious blinding eye disease. We reported three cases of fungal keratitis caused by Candida in 2011 in our hospital. It gave us a glance at the pathogen and the main features of the disease in our region.

    Culture preservation of medical fungi
    SANG Jun-jun, DENG Shu-wen, GUO Kai, PAN Wei-hua, LIAO Wan-qing
    2014, 9(2):  107-110. 
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    In recent years, mycoses have emerged as an important public health problem. The research about fungal epidemiology,pathogenesis,drug-resistant mechanism, prevention and treatment of mycoses is particularly important. Culture collections are responsible for providing the microbial resources for development of biological sciences. There are many preservation techniques of medical fungi. The choice of the method will depend to a large extent on the equipment and facilities available in the laboratory. The paper discusses quality management options and reviews different methods of fungal preservation, describing how the reproducibility and quality of these techniques are maintained in order to retain the full potential of fungi.

    Recent progress of polyamines in microorganisms
    YAO Xiang-wen, ZHOU Mi, CAO Ying-ying, JIANG Yuan-ying
    2014, 9(2):  114-118. 
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    Polyamines including putrescine, spermidine, spermine and cadaverine are polycationic molecules with hydrocarbon backbone and multiple amino groups. These subjects are essential metabolites presented in microorganisms. And polyamines involve many physiological processes of microbe, from growth to development and differentiation. Some studies found that polyamines could promote microbial cell proliferation, keep cell membrane, DNA and RNA stable, and play an important role in physiological stress, of late. This review focuses on the metabolism, transport and physiological functions of polyamines in microorganisms to provide more useful information.

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