Table of Content

28 February 2015, Volume 10 Issue 1

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Functional studies of neutrophils from phaeohyphomycosis patients with CARD9 deficiencies against Candida albicans

LIANG Pin, WANG Xiao-wen, WAN Zhe, LI Ruo-yu

2015, 10(1):  1-5. 

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Caspase recruitment domain-containing protein 9 (CARD9) is an adaptor molecule that is critical for NF-κB activation,and forms a complex with B-cell lymphoma 10 (BCL10) and mucosa-associated lymphoid tissue lymphoma translocation gene 1 (MALT1) that mediates C-type lectin receptors (CLRs)-triggered intracellular signaling during antifungal immunity.In the present study,we investigated the functions of polymorphonuclear neutrophils (PMNs) from phaeohyphomycosis patients with CARD9 deficiencies against Candida albicans (C.albicans).PMNs were isolated from patients and healthy blood donors and subsequently challenged with C.albicans.By calculating colony-forming units and MTT assay,the killing of C.albicans by CARD9-deficient neutrophils was detected,and the ROS production and phagocytotic ability of CARD9-deficient neutrophils was assayed by Flow CytoMetry (FCM).The phagolysosome ultrastructure and the secretion of IL-8 were analyzed separately by EM and CBA kit.As a result,we demonstrated that,compared with healthy donors,CARD9-deficient PMNs exhibited defects in C.albicans killing and IL-8 productions,which can be rescued in the presence of serum;however,the CARD9-deficient PMNs exhibited normal reactive oxygen species generation,phagocytotic ability and phagolysosome ultrastructure.In conclusion,our results indicate that CARD9 is indispensable for C.albicans killing by PMNs,and serum-opsonization acts as a CARD9-independent way,which could be a promising immunotherapy in the future.

Study of improved colony PCR technologies for the identification of Trichosporon asahii

ZHANG De-quan, LIU Han, LIAO Yong, XIA Zhi-kuan, YANG Dong-qian, YANG Yang, LV Xue-lian, YANG Rong-ya

2015, 10(1):  6-10. 

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Objective To explore a direct colony PCR method for rapid identification of T.asahii.Methods Four methods including direct colony,direct colony with centrifugation,enzymolysis,and improved enzymolysis method were used to preliminarily sample treatment.The sensitivity and the positive rate of colony PCR based on four methods for isolates of T.asahii were evaluated and compared with the PCR results of the traditional DNA extraction.Results The sensitivity of the improved enzymolysis method,direct colony method and the traditional DNA extraction were both 10² CFU/mL,while the positive rates were 93.75%,75% and 50%,respectively.The direct colony with centrifugation presented the sensitivity of 10⁴ CFU/mL and the positive rate of 43.75%.The enzymolysis method presented total negative results in the detection of both sensitivity and positive rate.Conclusion The improved enzymolysis method which presented the advantages of high sensitivity and positive rate of trace samples,could be applied for the rapid identification of T.asahii.

Clinical presentation and laboratory study for a case of cryptococcal meningitis caused by Cryptococcus gattii in Hebei province

DOU Hong-tao, WAN Zhe, YANG Qi-wen, WANG Yao, WANG He, XIE Xiu-li, LIU Juan, LI Ruo-yu, XU Ying-chun

2015, 10(1):  11-15. 

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Objective To report a case of meningitis caused by Cryptococcus gattii in North China and related laboratory investigation.Methods The strain isolated from the patient mentioned above was inoculated on the Chromagar media and Canavanine-glycine-bromthymol blue (CGB)media at 35℃.API 20C AUX、Vitek 2 Compact、Matrix-Assisted Laser Desorption Ionization-Time-of-Flight Mass Spectrometry (MALDI-TOF MS)、ribosomal ITS and IGS sequence analysis were performed for identification.Antifungal susceptibility test was also performed with ATB FUNGUS 3.Results The color of the strain on the Chromagar media was colorless and slight pink.It turned the CGB media blue five days later.It was identified as Cryptococcus neoformans by API 20C AUX、Vitek 2 Compact and MALDI-TOF MS.It was considered as Cryptococcus gattii by sequence analysis of genes of ribosomal ITS and IGS.The minimal inhibitory concentrations of 5-Flucytosine、AmphotericinB、Fluconazole、Voriconazole、Itriconazole against the isolate were ≤4 μg/mL、≤0.5 μg/mL、≤1 μg/mL、≤0.125 μg/mL、≤0.06 μg/mL respectively.Conclusions CGB media、ribosomal ITS and IGS sequence analysis can discriminate Cryptococcus gattii from Cryptococcus neoformans. CGB is an easy method to identify C.gattii.Isolation and identification of C.gattii need to be paid more attention.

Effects of polyubiquitin UBI4 downregulation on growth and pathogenesis of Cryptococcus neoformans

MENG Yun-fang, ZHOU Zhao-jing, YANG Ya-li, ZHAO Jing-yu, ZHANG Chao, FA Zhen-zong, SANG Jun-jun, FANG Wei, LIAO Wan-qing

2015, 10(1):  16-21. 

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Objective To construct Cryptococcus neoformans strain with downregulated UBI4 expression and examine the effects of UBI4 downregulation on growth and pathogenesis of C.neoformans.Methods UBI4 DNA fragment was amplified and infused into linear plasmid pCH20 containing histone promoter.The fused plasmid was then transformed into C.neoformans through gene gun.The positive transformants were screened by diagnostic PCR and DNA sequencing.Various phenotype of C.neoformans reconstituted strain H99^His-UBI4 were examined.Results Cryptococcus neoformans strain with low expression of UBI4 were constructed and phenotypic assays demonstrated that down-regulated UBI4 lead to slower growth rate and enhanced susceptibility to various stresses such as H₂O₂,NO and antifungal agents in C.neoformans.Moreover,C.neoformans with down-regulated UBI4 was also attenuated dramatically in wax moth infection model.Conclusion Our study suggested that polyubiquitin UBI4 played a critical role in cell proliferation,stress response and pathogenesis in C.neoformans.Our work laid a foundation for following studies about functions and mechanisms of ubiquitin system.

In vitro susceptibility testing of domestic terbinafine hydrochloride against dermatophytes

TAN Xin, PAN Wei-hua, DENG Shu-wen, GUO Yun, LEI Wen-zhi, ZHANG Chao, ZHOU Zhao-jing, FANG Wen-jie, WEN Yi-yang, SHEN Shuai-shuai, ZHU Shuang, YAO Zhi-rong, ZHENG Jie, DENG Dan-qi, LIAO Wan-qing

2015, 10(1):  22-24,5. 

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Objectives To investigate the in vitro susceptibility testing of terbinafine against 138 strains of dermatophytes.Methods CLSI M38-A2 guideline was applied.The media containing the fungi in the concentration of(0.4-5)×10⁴ CFU/mL were incubated at 30 for 7 days.The MIC endpoint was determined as the first concentration of antifungal agent that reached 100% inhibition of growth compared with the growth control.The MEC endpoint was determined as the first concentration of antifungal agent that reached 50% inhibition of growth compared with the growth control.Results The tested 138 strains of Dermatophytes were susceptible to terbinafine,MEC range:0.008-0.125 μg/mL,MIC 0.008-0.25μg/mL.There was no statistical differences between Beijia terbinafine and imported terbinafine(P>0.05).Conclusion Beija Terbinafine is active aganist Dermatophytes.It can be an effective choice in superficial mycosis treatment.

Anti-adhesion effect of Butyl alcohol extract of BaiTouWeng decoction on Candida albicans isolated from VVC

ZHANG Meng-xiang, XIA Dan, SHI Gao-xiang, LU Ke-qiao, SHAO Jing, WU Da-qiang, WANG Tian-ming, WANG Chang-zhong

2015, 10(1):  25-30. 

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Objective To investigate anti-adhesion effect of Butyl alcohol extract of BaiTouWeng decoction (BAEB)on Candida albicans.Methods Agar plate assay was used to determine CFU of Candida albicans for 2h and 4h.XTT reduction assay was applied to evaluate the adherent Candida albicans merabolic activity.Test tube assay was utilized to evaluate flocculation.The water-hydrocarbon two-phase assay was employed to measure the cell surface hydrophobicity of Candida albicans.Scanning electron microscope(SEM)and fluorescein diacetate(FDA)staining was used to observe the morphological changes of adherent Candida albicans.qRT-PCR was adopted to inspect the expression of adhesion-related genes such as INT1,ALS3,MNT2,ALA1,ALS1,ALS5,HWP1 and CSH1.Results 512,1 024 μg/mL BAEB could effectively inhibit adherence of Candida albicans.Flocculation was delayed by 512,1 024 μg/mL BAEB.The cell surface hydrophobicity in the BAEB treated groups was lower than that of the control group.The expression of ALS1,CSH1,ALA1,ALS3,INT1,HWP1 were downregulated dramatically after BAEB treatment,but ALS5 and MNT2 were unchanged.Conclusion BAEB could effectively inhibit adherence of Candida albicans isolated from VVC by regulating adhesion-associated genes.

Retrospective analysis of hepatitis complicated with zygomycosis in mainland China:a review of 19 cases

HU Zun-qi, ZHAO Jing-yu, FANG Wei, PAN Wei-hua, LIAO Wan-qing, CAI Qing-ping

2015, 10(1):  31-34. 

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Objective To investigate the epidemiology and clinical characteristics of hepatitis complicated with zygomycosis in mainland China.Methods Literatures about hepatitis complicated with zygomycosis in China were collected from the electrical databases,such as CNKI,CJDF,VIP,Chinese physician full-text journals,and also Pubmed since 2000.Results 10 articles (19 cases) were included in the study.Median age was 37.6 (range,20-62) years old,with a male predominance (M:F=18:1).Without specific clinical and radiographic features,its diagnosis was mainly dependent on mycologic and/or histopathological examination.Amphotericin B was the first-line antifungal drug for hepatitis complicated with zygomycosis,and better efficacy could be got if combined with surgical treatment.Conclusions In recent years,there is an increasing trend in the prevalence of hepatitis complicated with zygomycosis in mainland china.For highly suspected hepatitis complicated with zygomycosis,it's necessary to initiate early anti-fungal therapy,and combined surgical treatment may improve the outcome.

A case of infection caused by Aspergillus flavus after anterior cruciate ligament reconstruction

ZHOU Wan-qing, HONG Jun, ZHANG Zhi-feng, ZHANG Kui

2015, 10(1):  37-38. 

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This is a report of knee joint infection caused by Aspergillus flavus after anterior cruciate ligament reconstruction.A 26-year-old man was performed a surgery of cruciate ligament reconstruction with autologous tendon because of old cruciate ligament rupture.The symptoms of knee joint infection happened one week after the surgery and conventional anti-bacterial drugs showed ineffective.According to the history,clinical symptoms and the fungal culture positive with Aspergillus flavus,the patient was diagnosed as a fungal infection after surgery caused by Aspergillus flavus.After rinsing the joint cavity with 5% sodium bicarbonate solution and the use of amphotericin B (1% amphotericin B 20 mL,qd) infusion administration,combination use of voriconazole (200 mg,q12 h) oral for anti-fungal treatment,the patient's infection condition improved and was discharged after 28 days.

Dendritic cells in anti-fungal immunity

WANG Rui-li, LIAO Yong, AO Jun-hong

2015, 10(1):  39-43,21. 

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The number of life-threatening fungal infections has skyrocketed recently as a result of the abused of antibiotics,chemotherapy drugs and immunosuppressive drugs,as well as the increased immunocompromised patients.Dendritic cells(DCs)are the most powerful antigen-presenting cells.As the bridge between the innate and adaptive immune system,DCs play a central role in recognizing a variety of pathogens and presenting antigens.Many investigations demonstrate that DCs can effectively recognize fungal pathogens through several receptors on the cellular surface and promote the immune responses against invasive fungi.Here,we describe the characteristics of the various DCs subsets,the way of DC recognizing fungi,and their functions in immunity against fungal pathogens.

The research progress on antifungal resistance mechanisms of Candida spp.

HE Xiao-yuan, ZHAO Ming-feng

2015, 10(1):  49-53. 

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In recent years,the incidence and mortality of fungal infection,especially the opportunistic fungal infection have been increasing.This is mainly associated with antifungal resistance and the restricted number of available antifungal drugs.Candida species are opportunistic fungi in human body.The common antifungal resistance mechanisms include the changes of the target enzyme,the lack of target site,the drug permeability decline of fungal cell membrane,the increased expression of multi-drug resistance protein,the change of cholesterol synthesis bypass,the biofilm formation and so on.The latest literature reports that secreted aspartic proteinases have a role in antifungal resistance of Candida spp..In this article,the epidemiology of Candida spp.infection and its possible antifungal resistance mechanisms are reviewed briefly.

Research progress in the pathogenesis of Candida albicans

ZHU Dan-ting, LI Jing, YU Li-ying

2015, 10(1):  54-58. 

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Candida albicans is one of the most common fungal pathogens of humans that colonize the skin and mucosal surfaces of the healthy hosts. Candida albicans can cause the widespread infection in the skin mucous membrane and viscera of the hosts.This microorganism is the single cell pseudohyphae yeast,which can grow as both of yeast and filamentous forms in the host,and viscerarmed fungal dimorphism or,more precisely,fungal polymorphism.The relative attributes of the yeast and filamentous forms of Candida albicans,during the colonization of skin and mucosa,and later in the invasion of the blood stream and deep tissues,had long been debated.The pathogenesis of Candida albicans had been attributed to multi-factors,including adherence to host cells,secretion of degradative enzymes,morphology switching,successful biofilm formation,quorum sensing and so on.In this paper,the recent research progress in the pathogenesis of Candida albicans was reviewed.

Advances in external treatment of tinea pedis by traditional Chinese medicine

LI Su, FANG Liang-zhu, FAN Bin

2015, 10(1):  59-61. 

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Tinea pedis which caused by dermatophytes is called“Foot moisture”in Chinese medicine.In recent years,the treatment of tinea pedis by traditional Chinese medicine had showed better curative effects.This paper evaluated the methods of external treatment by traditional Chinese medicine and their curative effects.