Chinese Journal of Mycology

Effects of transforming growth factor on the function of macrophage against Cryptococcus neoformans

LI Ying-Fang, SANG Jun-Jun, LI Juan, JIANG Wei-Wei, LIAO Wan-Qing, PAN Wei-Hua

2017, 12(2): 65-68.

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Objective To investigate the phagocytosis and killing ability of macrophage under TGF-β intervention.Methods Using TGF-β intervene phagocytosis and killing assay of activated macrophages in vitro.Phagocytic index of macrophage and the intracellular growth of Cryptococcus neoformans was observed respectively.Nitric oxide (NO) production was detected using the Griess regent kit, and then comparing the changes of NO production in macrophages under the TGF-β intervention.Results The phagocytic index with the treatment of TGF-β decreased by 43.9%, significantly lower than control group (P<0.05).While the production of NO increased and the intracellular fungal burden significantly declined under the intervention of TGF-β (P<0.05).Conclusion Chemotaxis and phagocytosis of activated macrophage were inhibited under the intervention of TGF-β in vitro, but the NO production increased and killing ability enhanced.

Phenotypic characterization and virulence of two white nonsporulating Aspergillus fumigatus isolates

JIANG Yuan, ZHANG Cai-yun, WANG Feng-yuan, KONG Qing-tao, ZHANG Zheng, ZENG Qiu-qiong, LONG Nan-biao, SANG Hong

2017, 12(2): 69-73.

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Objective In this study, another white nonsporulating A.fumigatus strain (named as A2j) was compared with reported nonsporulating strain A1j in terms of morphology and virulence.Methods A2j was confirmed by DNA sequencing of internal transcribed spacer (ITS) regions and partialb-tubulin genes.We incubated A1j and A2j through the slide culture method to compare them under a microscope.We observe the morphology of A1j and A2j at different times or on various medium types.An immunosuppressed mice model was created to compare the virulence of A1j, A2j and Af293.Results A2j was accurately identified as A.fumigatus by ITS and β-tubulin sequencing.Both A1j and A2j showed abnormal germination under the microscope compared to those of the reference strain Af293 by the slide culture method.Neither increased time nor different medium types could stimulate the formation of spores and pigment.A1j cannot grow and A2j can grow at 50℃.A1j is hypovirulentin immunosuppressed mice model;however, A2j is virulent compared to the standard strain Af293.Conclusion The newly A.fumigatus A2j and A1j have significant differences in growth rate, thermotolerance and virulence.

Evaluation of ITS, BenA and CaM in identification of clinical Aspergillus

LI Ying, XU Ying-chun

2017, 12(2): 74-77.

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Objective To evaluate the internal transcribed spacer (ITS), β-tubulin gene (BenA) and calmodulin gene (CaM) sequence analysis in identification of clinical Aspergillus.Methods All 169 strains of Aspergillus collected from clinical patients were analyzed by ITS, BenA, CaM sequencing to obtain the species information with reference to the Genbank database.Results The tested strains were identified to species level with identification rates of 52.7% (89/169), 66.3% (112/169), 97.6% (165/169) and to genus level of 47.3% (80/169), 33.7% (57/169), 2.4% (4/169) by ITS, BenA and CaM, respectively.None of the three sequence markers failed in identification of Aspergillus.Conclusion ITS, BenA and CaM can be used to identify clinical Aspergillus.Among them, CaM is the most efficient molecular marker for Aspergillus identification.

Synergistic effect of mangiferin in combination with fluconazole against fluconazole-resistant Candida albicans

DONG Huai-huai, WANG Yuan-hua, LIAO Ze-bin, JIANG Yuan-ying, CAO Ying-ying

2017, 12(2): 78-82,85.

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Objective To explore the synergistic effect of mangiferin in combination with fluconazole against fluconazole-resistant Candida albicans.Methods The minimal inhibitory concentrations (MIC₈₀) of 22 fluconazole-resistant Candida albicans strains by mangiferin in combination with fluconazole were determined by checkerboard microdilution assay.Time-kill curves and cell growth test were used to investigate the synergistic effect on Candida albcians growth process.The expression of CDR1, CDR2, MDR1 were measured by Real time RT-PCR.Results Mangiferin in combination with fluconazole could inhibit the growth of Candida albicans obviously (FICI<0.5).Mangiferin and fluconazole-treated cells expressed lower level of CDR1 mRNA than the cells grown in the presence of fluconazole or mangiferin alone.Conclusions Mangiferin possessed a synergistic effect with fluconazole against fluconazole-resistant Candida albcians.

Antifungal susceptibilities of ten kinds of traditional Chinese herb extracts against Candida in vitro

LU Xiang-ting, HUANG Xin

2017, 12(2): 83-85.

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Objective To observe the antifungal effect of 10 kinds of traditional Chinese herb extracts in vitro.Methods We performed in vitro drug susceptibility testing for ten kinds of traditional Chinese herb extracts, fluconazole and itraconazole against Candida isolates following CLSI M27-A3 guidelines.Results Panax notoginseng saponins (PNS) and ginseng stem-leave saponins (GSLS) were more sensitive to C.albicans.Their 24 h MIC values were 62.5 μg/mL and the 48 h MIC values were 125 μg/mL.PNS and Citric acid were more sensitive to C.krusei at 24 h and their MIC values were both 1.96 μg/mL.PNS was more sensitive to C.krusei at 48 h, the MIC were 31.25 μg/mL.GSLS and Thymolwere more sensitive to C.parapsilosis and their 24 h MICs were both 3.91 μg/mL.GSLS was sensitive to C.parapsilosis at 48 h, the MIC was 15.63 μg/mL.Conclusion PNS, GSLS, Citric acid and Thymol possessed activity against Candida isolates in vitro.There were some differences in antifungal activity of different extracts of Chinese herbs against different kinds of Candida species.The results provide a theoretical basis for further screening of Chinese herb extracts.

Clinical values of (1, 3)-β-D-glucan in cerebrospinal fluid of patients with fungal meningitis

WANG Xing, YANG Liu, HE Xi, ZHANG Min

2017, 12(2): 86-88,91.

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Objective To evaluate the utility of (1, 3)-β-D-glucan (BDG)as an assist diagnostic tool for patients infected with fungal meningoencephalitis.To investigate the change of BDG level following the administration of antifungal drug in fungal meningoencephalitis patients, evaluate the monitor value of BDG in cerebrospinal fluid (CSF) for fungal meningoencephalitis.Methods BDG concentrations of 10 cerebrospinal fluid (CSF) specimens from confirmed cases of fungal meningoencephalitis, and 30 negative control CSF specimens were tested.And we also assessed whether BDG levels correlate with clinical status by using incident samples from 10 patients with serially collected CSF.Results The level of BDG in CSF was obviously rise in those confirmed cases of fungal meningoencephalitis, the data had a statistically significant difference (P<0.05).Patients with serially collected CSF displayed that patients with a decline in CSF BDG had clinical improvement.Conclusions Our data suggest that measuring BDG in CSF is a highly sensitive test for diagnosis of fungal meningoencephalitis.Analysis of BDG levels in serially collected CSF demonstrated that BDG may correlate with clinical response and course of treatment.Routine measurement of BDG in CSF may provide useful adjunctive data for the clinical management of patients with meningoencephalitis.

Colonization or infection of Scedosporium in lower respiratory tract of secondary pulmonary tuberculosis patients

ZENG Jing-si, LI Hong-jie, MAO Ye-hong, LIAN Xin, ZHENG Yue-chen

2017, 12(2): 89-91.

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Objective Understanding colonization or infection of Scedosporium in lower respiratory tract of secondary pulmonary tuberculosis.Methods Major pathogens of Scedosporiosis, 5 Scedosporium spp.(S.apiospermum, S.boydii, S.minutispora, S.aurantiacum and S.dehoogii), were detected in bronchoalveolar lavage fluid from the patients with secondary pulmonary tuberculosis using direct microscope examination, fungal culturing and PCR-based reverse line blot (PCR-RLB) targeting the partial β-tubulin gene.Result No Scedosporium spp.were found in 101 specimens of bronchoalveolar lavage fluid from secondary pulmonary tuberculosis patients.Conclusion Rare Scedosporium spp.colonize or infect in lower respiratory tract of secondary pulmonary tuberculosis.It implies that secondary pulmonary tuberculosis complicated by Scedosporiosis due to increasing colonization of Scedosporium spp.in the respiratory tract will be uncommon.

Administration of voriconazole in disseminated penicilliosis marneffei: a retrospective study

WEI Jin-ying, OUYANG Yan-yin, CAI Shuang-qi, LIANG Hao, CAO Cun-wei

2017, 12(2): 92-97.

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Objective The aim of this retrospective study is to evaluate the efficacy and safety of voriconazole in the treatment of Penicilliosis marneffei (PSM)patients with or without HIV infection in clinical setting.Methods The disseminated PSM patients who received intravenous voriconazole (6 mg/kg every 12 hours for the first 24 hours and then 4 mg/kg every 12 hours afterwards) as primary antifungal treatment were enrolled.The following duration of antifungal treatment varied at the discretion of investigators according to the response of the patients.Primary response was evaluated at Week 16 or at the end of treatment (EOT).Follow-up evaluations were performed at 6 months and 1 year after EOT.Results A total of seventeen patients enrolled in this study, three patients were not evaluable because the treatment discontinued prematurely.Among the remaining fourteen patients, thirteen had satisfied response to treatment, ten patients had complete response and three patients had partial response at Week 16.Only one patient was evaluated as failure.Follow-up assessments in eleven patients showed eight patients were cured and the remaining three patients relapsed at 6 months after EOT.Eight patients were assessed 1 year later, none of them relapsed.No adverse events associated to voriconazole were recorded during the treatment.Conclusion The results from our study suggest that voriconazole is an effective, well-tolerated therapeutic option for disseminated P.marneffei infection.

Causative agents of tinea capitis isolated from the patients in Hubei province and surrounding area in China during 1960-2015

MAO Ye-hong, HE Meng-wen, ZHENG Yue-chen, LIAN Xin, ZENG Jing-si

2017, 12(2): 98-101,105.

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Objective To understand the pathogens of tinea capitis during the period 1960-2015.Methods Fungal strains isolated from the patients with tinea capitis in the Medical Mycology Clinical Laboratory were subjected to a classification and statistical analysis.Morphological, physiological and biochemical methods were applied for fungal identification.Results Tinea capitis prevalence was high in the 1960s, with Trichophyton violaceum (41.9%) and Trichophyton schoenleinii (31.0%) being the most common pathogenic species, while the cases decreased sharply in the 1970s.Later in the 1990s, the proportion of Microsporum canis dropped to 1% after its peak (36.9%).T.violaceum had replaced T.schoenleinii and become the most common species (63%).T.schoenleinii and Trichophyton rubrum tended to infect adults, while T.violaceum, M.canis, Trichophyton interdigitale and Microsporum ferrugineum were more likely to infected children.The proportion of female patients among adults (64.6%) was higher than that of children (50.6%) (T.schoenleinii excluded) (P<0.05).Conclusion The pathogens of tinea capitis changed over time and now T.violaceum had become the most common species in the district.

Clinical infection characteristics and analysis of drug resistance trend of candiduria in hospital

CAO Yan-yun, FANG Hua, CAI Hai-bin, XU Shun-ming

2017, 12(2): 102-105.

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Objective To investigate and analyze the clinical infection characteristics and drug resistance trend of candiduria, to provide evidence for selection of antifungal drugs.Methods Patients who cultured Candida positive in urinary sediment were collected from 2013 to 2016.The strains were identified by CHRO Magar chromogenic medium and the drug susceptibility test was performed with ATB Fungu 3 reagent.Results A total of 202 strains of Candida were isolated, including C.albicans (79.1%), C.tropicalis (10.7%), C.parapsilosis and C.krusei (respectively 5.1%).The Candida were mainly found in emergency department (83 cases), followed by urology (21 cases) and endocrinology (21 cases).Drug sensitivity results showed that Amphotericin B and 5-fluorocytosine had a strong antibacterial activity against Candida.The resistance rate of Candida to fluconazole increased year by year.Candida's resistant to voriconazole and itraconazole was in a fluctuating state, and the resistance rate of Candida was significantly higher in 2016.Conclusion C.albicans was still the most predominant pathogen of candiduria.The drug resistance rates of amphotericin B and 5-fluorocytosine were the lowest, while there was a genus difference in drug resistance of the azoles, and the trends of drug resistance were becoming more serious.

A case report of disseminated mucormycosis

SONG Min, JIANG Lu-ning, SU Na, JIANG Sheng-hua, LIU Yan

2017, 12(2): 108-110,113.

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A 44-year-old female patient was hospitalized with cough, expectoration, headache and fever over ten days;chest computed tomography (CT)examination showed multiple double-pulmonary circular nodules and cavities with different sizes and clear boundary.The maximum cavity diameter was about 1.8 cm, and the density of soft tissue was found in the part of the cavity.Head magnetic resonance imaging (MRI) combined with contrast-enha showed the bilateral frontal lobe, left temporal lobe, left parietal occipital lobe lesion and cerebral ventricle lesions were annular and patchy.CT-guided fine-needle percutaneous biopsy was performed.The biopsy showed many necrotic granulation tissue fungal hyphae was found.The case was diagnosed as DM.Intracranial lesions were improved after treatment with empirical anti mucormycosis treatment.

The transcription factors of phenotypic transition signal pathways in Candida albicans

WANG Li-hong, ZHANG Jin-yu, YAN Lan, JIANG Yuan-ying, CAO Yong-bing, HONG Hao

2017, 12(2): 114-119.

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Candida albicans, responsing signals to adapt to environment by a variety of signal transduction pathways, is the most common human opportunistic fungi.The most important signal transduction pathways that mediate the morphogenesis and virulence in Candida albicans are cAMP-PKA pathway and MAPK pathway.Transcription factors are the endpoints of these pathways, such as Efg1 and Cph1.Transcription factors can regulate the transcription of genes by binding with their promoters in Candida albicans.Regulating transcription factors can promote or inhibit the signal transduction, and thus affecting the proliferation, morphogenesis, virulence, etc, of Candida albicans.In this review, we demonstrated the major transcription factors of phenotypic transition signal pathways in Candida albicans to further understand the phenotype and regulation mechanism of Candida albicans.

Human genetic susceptibility to Candida infections

KAN Si-yue, LI Xiao-fang, LIU Wei-da

2017, 12(2): 120-123.

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Candida spp.are medically important fungi causing severe mucosal and systemic infections, however, not all individuals are at risk to develop Candida infections.With the completion of the human genome project and the development of molecular genetic technology, the genentic susceptibility to Candida infections has attracted the attention of more and more researchers.Severe fungal infections are associated with monogenic primary immunodeficiencies such as defects in STAT1, STAT3 or CARD9.Furthermore, some polymorphisms in genes of the immune system have also been associated with fungal infections such as recurrent vulvovaginal candidiasis and candidemia.The discovery of the genetic susceptibility to Candida infections can lead to a better understanding of the pathogenesis of the disease, as well as the development of novel detection index and therapeutic target.

The resistance mechanisms of the biofilm related Candida albicans

MENG Ling-ning, LIU Jin-yan, LI Wen-jing, ZHAO Yue, XIANG Ming-jie

2017, 12(2): 124-127.

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Candida albicans can form biofilms on medical implant materials, leading to high mortality infection.Mature biofilm related C.albicans infections are difficult to cure because of multi-drug resistance.This review discusses the resistance mechanisms of biofilm related C.albicans from efflux pump genes and matrix and stress response, summarizing the research advance of the mechanism and providing reference for clinical prevention strategies.

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