Chinese Journal of Mycology

Detecting mannan-binding lectin and T-lymphocyte subsets for the diagnosis of pulmonary aspergillosis

YANG Yan, TENT Ge-ling, WU Yan-hua, ZHANG Lin-lin, GE Li-zhen, CHEN Ying, HU Hua

2017, 12(6): 328-331,336.

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Objective To investigate the role of mannan-binding lectin (MBL) level and changes of T-lymphocyte subsets on the patients with pulmonary aspergillosis.Methods From May 2013 to May 2015,51 patients were enrolled into disease group,52 healthy adults were enrolled into normalcontrol group.The levels of MBL and Galactomannanwere detected by Enzyme-linked Immunosorbent Assay (ELISA).The percentages of CD3⁺CD4⁺T cells,CD3⁺CD8⁺T cells,and the ratio of CD3⁺CD4⁺/CD3⁺CD8⁺T cells were determined by flow cytometer.Results The express of MBL in disease group (197.96±148.16 μg/ml) was significantly higher than that of normal group (120.25±98.65)(P<0.05).The serum galactomannan (GM) level of disease group (0.94±0.77 μg/L) was significantly higher than that of normal group (0.32±0.16 μg/L).The percentages of CD3⁺CD4⁺T cells and CD3⁺CD8⁺T cells were (33.07±7.9)%,(40.32±7.30)% (P<0.05),(33.00±8.29)%,(25.98±6.65)%(P<0.05)in disease group and normal group,respectively.The ratios of CD3⁺CD4⁺/CD3⁺CD8⁺T cells in disease group and normal group were 1.09±0.47,1.68±0.65 (P<0.05),respectively.Conclusion There was significant change of MBL level and the percentages of T-lymphocyte subset in patients with pulmonary aspergillosis compared with control group.Immunotherapy may improve the efficacy of pulmonary aspergillosis.Through the change of T-lymphocyte subset can preliminary assessment the body's immune status in patients,at the same time,providing a theoretical basis for immune enhancement therapy in pulmonary aspergillosis.

Transcriptome analysis of Candida biofilm formation exposed to Ultraviolet A

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

2017, 12(6): 332-336.

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Objective To explore the genetic expression of hyphal growth and biofilm formation of Candida albicans exposed to ultraviolet A using the transcriptome analysis.Methods In vitro Candida albicans (ATCC90028) biofilms were irradiated by a new type of LED-UVA antimicrobial device (wavelength:365 nm,intensity:13 mW/cm²,frequency:100 Hz).The differentially expressed genes between the irradiated groups and the control group were detected by RNA-Seq technology analysis and GO,KEGG enrichment analysis of genetic differences.Results A total of 1221 gene expression levels exhibited a significant difference,including 862 up-regulated genes,359 down-regulated genes.The expressions of MCR1,CBR1,MUQ1 and SHP1 were significantly increased.The expressions of RIO2 and MPP were significantly decresed.Conclusion UVA could induce apoptosis of Candia albicans by un-regulating the expression of MCR1,CBR1,MUQ1 and SHP1 and inhibit hyphal growth and development by un-regulating the expression of RIO2 and MPP.

Butyl alcohol extract of BaiTouWeng decoction inhibit vulvovaginal candidiasis in murine model via down-regulating NLRP3 inflammasome

DUAN Qiang-jun, YUN yun, CHEN Zhi-zhen, SHAO Jing, WANG Ting-ming, WANG Chang-zhong

2017, 12(6): 337-341.

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Objective To study the effects of Butyl alcohol extract of BaiTouWeng decoction (BAEB) on vulvovaginal candidiasis (VVC) in murine model.Methods Seventy-two female mice were randomly divided into normal group,model group,BAEB high-dose group,BAEB middle-dose group,BAEB low-dose group,and fluconazole group.The mice were firstly injected with estradiol to induce oestrum,and then intravaginally administrated with 2×10⁶ CFU/mL Candida albicans.After 7-day treatment,The fungal load (CFU/mL) was assessed from vaginal lavage fluid,the pathological changes of vaginal tissues were observed by HE staining.The levels of IL-1β and IL-18 in vaginal tissues were detected by ELISA.The mRNA expressions of NLRP3,ASC,caspase-1 were analyzed by qRT-PCR.Results Compared with the normal group,the vaginal mucosa of the model group was completely destroyed,and the infiltrations of plenty of inflammatory cells were observed.Compared with the model group,BAEB groups showed improvements to varying degrees.The levels of IL-1β and IL-18 in the model group were significantly higher than those in the normal group (P<0.001).The mRNA expressions of TLR2,TLR4,NLRP3,ASC and caspase-1 were up-regulated significantly (P<0.001).The levels of IL-1β and IL-18 in the BAEB groups were much lower than those in the control group (P<0.001).Conclusion The BAEB could be used to treat the vulvovaginal candidiasis in mice.

Quantitative levels of anti-Pneumocystis jirovecii major surface glycoprotein IgG and IgM antibodies in HIV and non-HIV groups

GUO Shuang, LIU Yi, TIAN Xiao-jun, LI Shao-gang, SONG Ying-gai, REN Yi

2017, 12(6): 342-345.

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Objective To investigate the quantitative levels of anti-Pneumocystis jirovecii (Pj) major surface glycoprotein (Msg) IgG and IgM antibodies in HIV and non-HIV groups to explore its value in epidemiological investigation.Method One target sequence of Pj Msg gene was selected and expressed using prokaryotic expression system.Enzyme-linked immunosorbent assay (ELISA) was established to detect anti-Pj Msg IgG and IgM antibodies using recombinant Pj Msg as antigen.Multiple proportion dilution methods were established to measure quantitative levels of anti-Pj Msg IgG and IgM antibodies.A cohort of individuals with HIV infection (n=176) and non HIV controls (n=266) in Beijing China were investigated.All patients were not diagnosed with a situation of pneumocystis pneumonia (PCP).Results The positive rate of anti-Pj Msg IgG antibody in non-HIV group was 42.5% (113/266).Among these 113 positive cases,there were 77 cases (68.1%),15 cases (13.3%),6 cases (5.3%),and 15 cases (13.3%) in the titer of 1:100,1:200,1:400,and 1:800 levels respectively.The positive rate of anti-Pj Msg IgG antibody in HIV infection group was 24.4% (43/176).Among these 43 positive cases,there were 28 cases (65.1%),10 cases (23.3%),3 cases (7.0%),and 2 cases (4.7%) in the titer of 1:100,1:200,1:400,and 1:800 levels respectively.There was no statistical significance in the difference of anti-Pj Msg IgG antibody titer levels compared with non-HIV controls (χ²=4.254,P=0.235).The positive rate of anti-Pj Msg IgM antibody in non-HIV group was 41.7% (111/266).Among these 111 positive cases,there were 57 cases (51.4%),21 cases (18.9%),19 cases (7.1%),and 14 cases (12.6%) in the titer of 1:100,1:200,1:400,and 1:800 levels respectively.The positive rate of anti-Pj Msg IgM antibody in HIV infection group was 12.5% (22/176).Among these 22 positive cases,there were 10 cases (45.4%),2 cases (9.1%),4 cases (18.2%),and 6 cases (27.3%) in the titer of 1:100,1:200,1:400,and 1:800 levels respectively.There was no statistical significance in the difference of anti-Pj Msg IgM antibody titer levels compared with non-HIV controls (χ²=3.788,P=0.285).Conclusions Among these non-PCP individuals,the anti-Pj Msg IgG antibody titer levels were mostly between 1:100 and 1:200.The methods of antibody quantitative levels and their clinic significance need further studies.

Clinical analysis of invasive fungal diseases in 327 patients with hematological malignancies

ZHAO Wei, SUN Fang

2017, 12(6): 346-351.

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Objective To investigate clinical features and pathogenic distributions of invasive fungal diseases in patients with hematologic malignancies,analyze risk factors of the therapeutic effect and the usage of antifungal agents in order to provide reference for clinical diagnosis and treatment.Methods The clinical data of 327 cases of invasive fungal diseases in patients with hematologic malignancies,who were treated in the hospital from Jan 2014 to Dec 2015,were retrospectively analyzed,and the pathogenic distributions,infectious sites,therapeutic effect,risk factors and use of antifungal agents were statistical analyzed by the use of software SPSS 21.0.Results Totally 410 fungal strains were isolated from the 327 patients,including Candida albicans (80.73%) and Aspergillus (9.76%);and the lung was the main infection sites (80.81%).The total cure rate was 61.77%,and the mortality was 12.84%.The therapeutic effect was positively correlated with the patients treated with glucocorticoid and more than three broad-spectrum antibiotics;while there was a negative correlation between the age of patients and therapeutic effect.The irrational use of antifungal agents included lack of loading dose,dosage and drug interaction.Conclusion The Candida albicans is the predominant species of pathogens causing invasive fungal infections in patients with hematologic malignancies.The main risk factors are aging and application of glucocorticoid and broad-spectrum antibiotics.It is important to focus on the elderly patients,and to strengthen the reasonable applications of glucocorticoid and broad-spectrum antibiotics.The unreasonable use of antifungal agents is relatively common.Usage and dosage,and drug interaction should be noted,and the supervision of antifungal agents use should be strengthened.

The cinlical effency of caspofungin and voriconazole on fungal infections in severe burn patients

YIN Xi-Yang

2017, 12(6): 352-355.

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Objective To explore the clinical effency of caspofungin and voriconazole on patients with severe burn complicated with fungal infections.Methods The clinical data of 62 patients with severe burn complicated with fungal infectionsin our hospital from June,2012 to June,2016 were collected.They were randomly divided into two groups according random number table.30 subjects who were treated with caspofungin were assigned to the control group,while 32 subjects in the observation group were treated with voriconazole.The clinical characteristics of the 62 patients were analyzed,and the clinical effects and the drug safety of the two groups were compared.Results The clinical symptoms included fever,pulmonary rales,sore throat,cough and expectoration,mental disorders,lung shadow and rale.The main infection sites were wound infection.And Candida albicans and Candida tropical were the main strains of fungi,which accounted for 55.93%and 20.33%,respectively.The total effective rates of the observation group and the control group were respectively 84.38% and 86.67%,and the difference between the two groups had no statistical significance (P>0.05).In addition,the incidence rates of the adverse reaction of the two groups were low.Conclusion Caspofungin and voriconazole had good clinical effect with less adverse reactions in the treatment of fungal infection secondary to severe burn.

A rapid and efficient extraction method of fungus DNA for PCR

CHAI Hai-yun, ZHANG Lei, YUAN Mu-ge, ZHU Qing-yi

2017, 12(6): 359-361,367.

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Objective To explore a new convenient and efficient fungi DNA extraction method,which could be used in many kinds of fungi.Methods We associated enzymic digestion with Chelex-100 to make up a new reagent for fungi DNA extraction.Results During the experiment,34 strains were tested.The DNAs of 32 strains could be obtained by our reagent,while only 27 strains' DNAs could be extracted by Takara lysis buffer.For the strains we did not successfuly obtain the DNA by the above methods,we grinded the fungi by liquid nitrogen.After that,our reagent obtained their DNA by,and only one by Takara lysis buffer.Conclusion The DNA extraction efficiency of our reagent is much better than that of Takara lysis buffer.It can be used in many kinds of fungi.For some fungi,we should firstly grind it before DNA extraction.

First case of pseudomembraneous lesions of the scrotum due to Trichosporon asteroids in China

YIN Song-chao, LI Mei-rong, WEI Ling, HUANG Huai-qiu, FENG Pei-ying

2017, 12(6): 362-364.

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Objective To report a case of scrotal trichosporonosis presenting with powdery and pseudomembraneous lesions on the scrotum in an immunocompetent patient.Methods A 25-year-old man complained of white crust on the scrotum for 6 weeks.We confirmed the diagnosis by KOH mount of the skin scrapings,fungal culture,CHROM agar culture,urease test,cycloheximide and molecular verification.Results KOH mount of the skin scrapings was positive for septate hyphae.Trichosporon asteroides was isolated from the lesions and confirmed by morphological and molecular analysis,a basidiomycetous yeast showing meristematic as well as hyphal growth with arthroconidia.The isolate colonies were blue in color on CHROM agar Candida agar.The urease test was positive,and the isolate was resistant to cycloheximide.The patient responded well to the use of oral and topical terbinafine,and his lesions cleared completely after two weeks.Conclusions Scrotal trichosporonosis due to Trichosporon asteroids was rarely reported in clinic,study on this case will assist in diagnosis and treatment of cutaneouse trichosporonosis.

One rare case of ankle abscess caused by Cladosporium tenuissimum

ZHENG Wen-ai, XU He-ping, YANG Xian-xu

2017, 12(6): 365-367.

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One rare case of ankle abscess caused by Cladosporium tenuissimum was reported.A 68-year-old peasant woman developed swollen,abscess and ulcer at his right ankle after leech bite one year ago.Histopathological examination revealed brown chlamydospores in tissue and fungal culture showed olive filamentous fungi growth which was identified as Cladosporium tenuissimum by morphology and ITS sequencing.This patient was diagnosed as chromablastomycosis and was prescribed itraconazole 0.2 g bid.The follow-up of four weeks showed obvious improvement.

A case of fungal peritonitis caused by Exophiala dermatitidis

TENG Yuan-ji, YI Xue-li, LUO Bin, GU Xian-jun, WEI Ye-sheng, DENG Ying, WEI Ying, NONG Xue-juan

2017, 12(6): 368-370.

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Report a case of fungal peritonitis in CAPD patient caused by Exophiala dermatitidis.Ed mainly caused skin,subcutaneous tissue,central nervous infection,but rarely was found in human peritonitis infection.The analyzed of characteristic,molecule examination and antifungal susceptibilities of pathogen were carefully performed.The patient showed improved after CAPD was replaced by hemodialysis and oral itraconazole (0.2g/d) treatment.

Enhancing the monitoring of Sporothrix schenckii complex for biosecurity

HUANG Meng-ya, ZHOU Xun

2017, 12(6): 371-374.

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Revolutionary changes in the molecular biology and taxonomy of most fungal groups had resulted in the tremendous changes of numerous names and classification including quarantine pests.Sporothrix schenckii was regarded as the causative agent of sporotrichosis in China,which however,represented a complex comprising six species.The causal agent of sporotrichosis in China had been proved to be S.globosa and S.schenckii in several independent studies.Other species in the complex had not been discovered in China.Since these species present different characters on virulence and medicine sensitivity,it is a necessity to enhance the monitoring and preventing of these species from entering the country.

Developments of MALDI-TOF MS in identification of clinical filamentous fungi

LI Ying, XU Ying-chun

2017, 12(6): 378-381.

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Invasive infections caused by filamentous fungi have been increasing in recent years.Accurate identification of filamentous fungi isolates has important role in clinical treatment of filamentous fungi infections owing to the great variety of antifungal susceptibility profiles among different species of those microorganisms.Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) has been revealed to be an emerging technology in the field of microbe identification and performed outstandingly in bacteria and yeasts identification.The performance of this novel technology in filamentous fungi identification,however,remained inferior and less-employed than that in bacteria and yeasts identification.This paper reviews some representative research aiming on working this dilemma out in the global and the remarkable developments obtained during this process.

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