Table of Content

28 June 2013, Volume 8 Issue 3

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The application of long-fragment fusion PCR in the construction of rho1-compelement strain of Aspergillus fumigatus

HAN Gai-ge, JIA Xiao-dong, HAN Xue-lin, ZHAO shuai, HAN li

2013, 8(3):  129-133. 

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Objective To evaluate the effect of primer design and the amplification condition on long-fragment fusion PCR.Methods With the example of 6.5 kb-long fragment amplification by fusion-PCR during construction of rho 1 gene complement in Aspergillus fumigatus, primer design with diverse homologous regions and different condition for amplification were evaluated.Results The 6.5 kb-long fragment was successfully amplified by fusion-PCR under following conditions: the primers with homologous region of 35 bp; DNA polymerase with high amplification efficiency and high fidelity; the concentration of template fragments higher than 15 ng/μL in whole reaction system.Conclusion With the appropriate primer, DNA polymerase and concentration of templates, the fusion PCR for the long-fragment might be a highly-efficient tool in gene-manufacturing of filamentous fungi.

Construction and activity test of animal model of Cryptococcus neoformans biofilm

ZHOU Jie, PAN Wei-hua, LIAO wang-qing

2013, 8(3):  134-136. 

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Objective To construct a kind of simply operated Cryptococcus biofilm infection animal model in vivo.Methods Build Cryptococcus neoformans biofilm infection model by placing catheter in subcutaneous tissue of rats and observe the structure of biofilm with electron scanning microscope. To test the activity of biofilm in vivo by MTT method. Result Successful construction of Cryptococcus neoformans bioflim rat model was done. The activity of biofilm in vivo was tested by MTT which showed an increase as the time prolonged.Conclusion Construction of Cryptococcus neoformans bioflim rat model is simple, available and easy to be operated. In some degree, it takes on the meaning of practical implication.

Deletion and reconstruction of HOG1 gene in Cryptococcus gattii

HUANG You-ming, LOU Yong-hua, ZHU Hong-mei, WEN mei

2013, 8(3):  137-141. 

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Objective To construct the HOG 1 mutant strain and the HOG 1 reconstituted strain.Methods Cloning HOG 1 gene according to the gene sequences, we got dHOG 1 gene by deleting part of HOG 1 gene.Then the HOG 1 gene and dHOG 1 gene were subcloned into eukaryotic vectors pGAPzα-A respectively. Finally, pGAPzα-dHOG 1 was transferred into Crypcococcus gattii original strain to get HOG 1 mutant strain. By the same method, pGAPzα-HOG 1 vector was transferred into HOG 1 mutant strain to get HOG 1 reconstituted strain.Results Results of RT-PCR showed that the HOG 1 mutant strain did not express whole HOG 1 gene fragment, while HOG 1 reconstituted strain expressed intact HOG 1 gene.Conclusion Successful construction of HOG 1 mutant strain and HOG 1 reconstituted strain would be useful in further research of virulence and pathogenesis of Cryptococcus gattii.

Sterilization of Candida albicans by Atmospheric pressure non-equilibrium plasma

LIAN Xin, CHEN Juan, FENG Ai-ping, ZENG Jing-si, ZHENG yue-chen

2013, 8(3):  142-146. 

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Objective To investigate the sterilization effect of Atmospheric pressure non-equilibrium plasma (APNP) to Candida albicans and its mechanisms.Methods Standard strains of Candida albicans were treated with APNP with different gas flows, operation durations and distances. Growth of the strains was observed, CFU was calculated with plate counting method and the changes of cell structure were observed with electron microscope.Results The growth of Candida albicans strains was suppressed significantly after treated with APNP. The inhibitory effect was greater with the gas flow of 2/0.06 L/min (He/O₂) than with that of 2/0.02 L/min, the operation duration of 10 min was better than 2 min, and the distance of 1 cm was better than 2 cm. After exposure to APNP, the fungal cells showed wizened, broken cell wall and cell membrane, leakage of cell content under electron microscopes.Conclusion s The sterilization effect of APNP on Candida albicans is significant. This effect is promoted with more O₂, longer operation duration and nearer distance. The main mechanism may be attributed to the active particles, which cause the breaking of the cell wall and membrane.

Studies on the relationship between hemolytic activity and biofilm formation ability of Trichosporon asahii isolates and their genotypes

SUN Wei, SU Jian-rong, XU shu-zhen

2013, 8(3):  147-150. 

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Objective To investigate the in vitro hemolytic ability and biofilm formation capacity of Trichosporon asahii strains isolated from urinary tract infected patients and their relationship with corresponding genotypes. To provide laboratory basis for clinical diagnosis and treatment of Trichosporon asahii.Methods Glass beads method and PCR technology were used to extract total DNA and determine the specific genotypes of clinical isolates (primers for IGS1 area), respectively; Plate method and XTT reduction assay were carried out to detect the hemolytic activity and biofilm formation capacity of clinical isolates.Results Ten Trichosporon asahii strains isolated from urinary tract infections belonged to genotype Ⅰ, Ⅲ, Ⅳ, and genotype Ⅳ was the most prevalent. All isolates had varying degrees of hemolytic activity. Except one case isolate, the remaining isolates had the ability to form biofilms in polystyrene surface. Genotype Ⅲ strains had stronger hemolytic activity and biofilm formation capacity.Conclusion s Ten Trichosporon asahii isolates from urinary tract infection were mainly genotype Ⅳ, while genotype Ⅲ strains showed stronger hemolytic activity and biofilm formation ability among all strains.

The diagnostic value of plasma 1,3-β-D glucan to invasive fungal infection in organ transplant recipients

LIU Yao-ting, CHEN Xian-feng, HU Hai-qing, YING xing, CHEN yong, ZHOU Ting-yin, ZHONG ren-qian

2013, 8(3):  151-155. 

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Objective To evaluate the diagnostic value of plasma 1,3-β-D glucan (BG) assay (G test) to invasive fungal infection (IFI) in organ transplant recipients.Methods By retrospectively accessing the clinical data of pediatric inpatients who underwent G test.The patients being recruited from January 2011 to December 2012 were split into two groups: group IFI comprised 64 patients with proven and probable IFI. Group control comprised 66 patients without fungal infection. G test was performed by the GKT-5M Set Kinetic Fungus Detection Kit. It was judged as positive when the plasma BG level≥10 pg/mL. 2 by 2 table was used to calculate the G test for the diagnosis of IFI sensitivity, specificity, positive predictive value and negative predictive values. Receiver operating curve (ROC) of G test was analyzed, and the area under the curve was calculated.Results In IFI group G test was positive in 57 cases, the positive rate was 89.06%. 15 cases of non-IFI group were positive, false positive rate was 22.7%. The G test sensitivity, specificity, positive and negative predictive values were89.1%,77.3%,79.2% and 87.9% respectively. According to ROC analysis, the AUC was 0.875 (95%CI: 0.813~0.937).Conclusion s G test has good diagnostic value of IFI in organ transplant patients. Appropriately higher cutoffs or continuous monitoring may significantly control false positive rate of the test.

The CT features in 17 cases of allergic bronchopulmonary aspergillosis

ZONG Xiao-fu, WANG Hui, YU tong-fu

2013, 8(3):  156-159. 

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Objective To evaluate the CT features of allergic bronchopulmonary aspergillosis (ABPA), and to improve this problem.Methods The CT features in 17 cases of ABPA were analyzed retrospectively, all patients were diagnosed according to the standard diagnostic criterion of ABPA.Results The CT depicted central bronchiectasis in 17 cases.The mucoid impaction of the dilated bronchial tree were seen in 13 cases, presenting as gloved-finger and toothpaste and "Y" appearance.CT showed bronchiectasis and tree buds signs in all patients;patchy areas of consolidation or atelectasis were observed in 9 cases.Fleeting infiltration in 2 cases on following up.Conclusion s The diagnosis of ABPA should be considered in patient with long-standing asthma whose CT demonstrates central bronchiectasis with slightly hyperdense mucoid impaction.

The study of the lectin level in healthy population,vulvovaginal candidiasis (VVC) and recurrent vulvovaginal candidiasis (RVVC) groups

LIU Ge, WANG Wei-hua, SUN Jing, MA Xue-feng, CHEN ai-qun

2013, 8(3):  160-162,168. 

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Objective To value the levels of mannose lectin in healthy population, vulvovaginal candidiasis (VVC) and recurrent vulvovaginal candidiasis (RVVC) groups.Method All the VVC and RVVC cases were given standardized treatment. Compared with healthy population, the MBL levels before and after treatment were observed in these two groups.Results The MBL levels of vvc group,RVVC group,healthy group were arranged from high to low numerically. But there was no significant difference among groups or within every group before and after treatment(P>0.05). There were 14% (5/43) cases of recurrence in VVC group and 48% (13/30) cases of recurrence in RVVC group,respectively. There was no significant differecne of recurrence between VVC and RVVC groups(P>0.05).Conclusion The level of MBL is higher numerically in VVC group than the level in RVVC group. The recurrence in both groups is consistet with the previously literature. The VVC patients with MBL level higer than 0.8 ng/mL seems to have lower risk of suffering RVVC.

Epidemiology of zygomycosis in mainland China

WANG Si-bu, LI Ruo-yu, YU jin

2013, 8(3):  163-168. 

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Objective To understand the epidemiology of zygomycosis and the status of diagnosis and treatment in mainland China.Methods Literature from 1976 to 2010 were collected by CNKI and Pubmed and statistically analyzed.Results A total of 206 cases and 428 patients were included in our study. The most common infections of gastrointestinal, pulmonary, sinus and cutaneous sites were often secondary to diabetes, gastrointestinal ulceration, burns, surgery, trauma, or no underlying conditions. Etiology diagnosis depended on direct microscopy, culture and pathology, and only 6 cases identified by molecular biology. The antifungal therapy was the main treatment and amphotericin B was the first choice in most cases. The mortality was 29.43% (126/428).Conclusion s Zygomycosis in mailand China had emerged as an increasingly important fungal infection which was in concordance with the world. Gastrointestinal tract was the most popular involvement. Morphological and molecular biological identification are important for accurate and prompt diagnosis. In vitro susceptibility test is helpful for clinical treatment.

Development trend analysis of Chinese Journal of Mycology：based on bibliometrics

JIA Ze-jun, DENG Xiao-qun, LIU jian-min

2013, 8(3):  169-172,178. 

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Objective To analysis the publications of Chinese Journal of Mycology, and make clear the development trend of the journal, in order to guide the journal development.Methods The literatures of Chinese Journal of Mycology were searched in CNKI database (2006-2012) using the following terms:soures=(Chinese Journal of Mycology). The searching results were refined and checked manually. Then, the final searching results were analysis by the software based on CNKI.Results From the first issue in 2006 to 2012, 42 issues and 843 articles were publicated, about 20 articles per issue and 120 articles per year. The annual number of publication were steadily from 2006 to 2012. To June 9, 2013, the citation frequency was increasing steadily, the top one was 42 times and the number of the article with citation frequency>=4 was 169, about 20.05%, according with the Pareto principle. The combined citation frequence and comprehensive citation frequence were increasing significantly, and the comprehensive IF was increasing slightly, comprasing with the first issue. While, self-citation rate was high. The majority article source was limited in the facilities of chief-editor, deputy chief-editors. The basic research and TCM research on Mycology were the hotspots.Conclusion From 2006 to 2012, Chinese Journal of Mycology develop well, covering the major institute of Mycology, while the publications volume, citation frequency, impact factor, and funded articles ratio were lower, remaining further improvement.

Treatment effect of naftifine/ketoconazole cream to tinea cruris

LIU Lei, LI zhen-lu

2013, 8(3):  176-178. 

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Objective To observe the clinical efficacy in the treatment of tinea cruris by naftifine/ketoconazole cream.Methods One hundred and sixty patients with tinea versicolor were randomly divided into trial group and control group. Naftifine/ketoconazole cream were applied in the trial group. Miconazole nitrate cream were applied in the control group. Both were used in the morning and evening. Clinical and mycological therapeutic effects were evaluated in 2 weeks of treatment and in 3 weeks of drug withdrawal.Results The effective rates in the treatment group and control group were 92.5% and 76.25% respectively,there was significant difference between the two groups (P<0.05).Fungs clearance rate of the treatment group was 97.50% whereas control group was 87.5% at the end of 2 weeks treatment. There was significant difference between the two groups (P<0.05). The effective rates in the treatment group and control group were 96.25% and 75.00% respectively,there was significant difference between the two groups (P<0.05).Fungs clearance rate of the treatment group was 97.50% whereas control group was 85.00% at discontinued 3 weeks. There was significant difference between the two groups (P<0.05).No obvious adverse reaction.Conclusion Naftifine/ketoconazole cream in treatment of tinea cruris is effect and safe.

The research advance of the medical fungal genomics

ZHAN ping, WANG Chong, LIU wei-da

2013, 8(3):  179-184,191. 

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A serial of medical fungus genome data have been published in recent years, by which human can make a better understanding of the pathogenesis of these pathogens. Here we try to review the history of the medical fungal genomics,genome sequencing technology and introduce the development and progress of clinical important medical fungi genome.

Research progress of Chinese herbal medicine against Candida albicans

LI Shu-yi, XIA Zhi-kuan, YANG rong-ya

2013, 8(3):  185-187. 

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Candida albicans is one of the most common human fungal pathogen, which can cause a variety of superficial and deep mycoses,and often become resistant to commonly used antifungal agents. So looking for broad-spectrum, high efficiency, low toxicity anti-Candida drugs has became a hot research. Extremely rich resources of Chinese herbal medicine have certain advantages of the prevention and treatment of Candida infections. To study the role of Chinese herbal medicine against Candida albicans,articles summarize the three aspects: herbal mechanism of action against Candida albicans and its active ingredient, single herb and compound herbal preparations against Candida albicans,synergistic effect of compound traditional Chinese medicine and western medicine against Candida albicans.The Chinese herbal medicine against Candida albicans clinical and experimental studies in recent years were reviewed. The article further confirmed the role of herbal anti-Candida albicans and showed the broad application prospects of herbal antifungal aspects.