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Table of Content

    28 June 2019, Volume 14 Issue 3
    Original articles
    Construction of RNA-interfering strains of Sporothrix schenckii STE20 gene
    ZHAO Lin, WANG Lin-hui, ZHENG Fang-liang, HOU Bin-bin
    2019, 14(3):  129-133. 
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    Objective This study aimed to construct a universal RNA interference vector of fungi, and to be widely used in the research of gene interference of various fungi. Methods Using the plant double expression vector, multi-cloning site (MCS) of pBlueScriptⅡ was cut down to construct our interference vector. To make the vector suitable for Agrobacterium transformation, we added t-DNA boundary repeats. In order to facilitate screening after transformation, hygromycin resistance genes were constructed into interfering vectors. In order to expand the interference range, we amplified the universal promoter ptrpc, spacer sequence and terminator Ttrpc in psilent-1 plasmid, and introduced hygromycin resistance gene to construct the vector. Results The vector PCB309-PSUST constructed in this experiment and the optimized reaction system were successfully used to effectively interfere with the STE20 gene of Sporothrix, and the operation was simple, the transformation efficiency was high, and the transformants were stable. Conclusion The constructed vector is suitable for Agrobacterium tumefaciens mediation and can be used for RNA interference research of filamentous fungal genes.

    Retrospective analysis of cerebral aspergillosis in mainland China from 2000 to 2019
    XUE Qiang, LEI Yan, ZHANG Chao, LIAO Wan-qing, HOU Li-jun
    2019, 14(3):  134-140. 
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    Objective To help early diagnosis and efficacious treatment of central nervous system(CNS) aspergillosis, we reviewed 39 cases in China since 2001. Methods We collected and analyzed retrospectively the information of patients' sex and aged composition, geographical features, clinical manifestations, associated diseases, therapeutic methods and outcomes. Results CNS aspergillosis could involve immunocompromised patients and immunocompetent patients, predominantly in middle aged men. The infection of CNS often resulted from the dissemination of the Aspergillus from lungs and paranasal sinuses. There were no early specific symptoms. The chief complaint was usually the symptoms of increased intracranial pressure, with half fever(51.28%). The combination of antifungal therapy with neurosurgery seems to be the most efficacious approach, and voriconazole was the best one. This disease had a poor prognosis, with high mortality. Conclusions With atypical clinical symptoms, nonspecific laboratory or imaging findings, and difficult diagnosis, CNS aspergillosis was still a disease with high mortality.

    Evaluation of the in vitro antifungal activity of the domestic caspofungin against clinical Aspergillus isolates
    YU Shu-ying, ZHANG Li, LI Ying, XU Ying-chun
    2019, 14(3):  141-146. 
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    Objective To evaluate the in vitro antifungal activity and consistency of original and domestic caspofungin against clinical Aspergillus isolates. Methods A total of 75 clinical Aspergillus isolates were collected from Peking Union Medical College Hospital. The antifungal susceptibility testing was carried out using the microdilution method recommended by CLSI. Results The MEC values of echinocandins were generally lower than the MIC values of the azoles, while MEC50 and MEC90 of micafungin were lower two dilutions than that of caspofungin. Except for one Aspergillus flavus isolates with MIC of 4 μg/mL, the MICs of remaining 74 Aspergillus isolates to amphotericin B were no more than 2 μg/mL. All the Aspergillus isolates were susceptible to voriconazole and posaconazole, but 16 Aspergillus fumigatus, 2 Aspergillus flavus and 5 Aspergillus niger isolates were resistant to itraconazole. The MEC values of original and domestic caspofungin against 11 Aspergillus isolates were inconsistent, but the MEC values of domestic caspofungin were only lower one dilution than those of original caspofungin. The consistency occurred in the antifungal susceptibility of over 76% of Aspergillus isolates to caspofungin and micafungin between 24h and 48h results. Conclusions Compared with azoles and amphotericin B, caspofungin had good in vitro antifungal activity against Aspergillus isolates. and domestic caspofungin was highly consistent with original caspofungin.

    Inhibitory effect of extract of Sophorae Flavescentis Radix-Cnidii Fructus Couplet medicines against Candida albicans isolate from VVC
    SHI Gao-xiang, JIANG Jing-jing, WANG Yun-xia, ZHAO Ting, SHAO Jing, WANG Tian-ming, Wang Chang-zhong
    2019, 14(3):  147-153. 
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    Objective To investigate the antifungal activity of extract of Sophorae Flavescentis Radix-Cnidii Fructus Couplet medicines against Candida albicans. Methods Microbroth dilution method was performed to determine the minimal inhibitory concentration (MIC). XTT reduction assay was used to evaluate the metabolic activity of C. albicans. Inverted microscopy and scanning electron microscopy (SEM) were used to observe C. albicans yeast-hypha conversion and biofilms respectively. Quantitative real time PCR (qRT-PCR) was performed to measure the expression of hypha and biofilms-related genes. Results ESCC showed significant antifungal effect and the MIC80 range was 512~1024 μg/mL. The metabolic activity of C. albicans was inhibited by ESCC, and induced incomplete three-dimensional structure of C. albicans biofilms. And expression of hypha and biofilms related genes was down-regulated with ESCC exposure. Conclusion This study demonstrated that ESCC showed prominent anti-fungal activity via affecting yeast-hypha conversion and biofilms of C. albicans.

    Clinical observation of long-pulsed 1064nm laser combined with ketoconazole ointment in the treatment of onychomycosis
    CAO Yan-yun, SUN Ying-fei, XU Yang, KONG Wei, XU Shun-ming
    2019, 14(3):  154-158. 
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    Objective To observe the clinical efficacy of long-pulsed 1064nm laser combined with ketoconazole ointment in the treatment of onychomycosis. Methods Seventy-three outpatients with onychomycosis were randomly divided into two groups:the combined group (37 cases, 75 affected nails) was treated with 1064nm long-pulsed laser combined with ketoconazole ointment encapsulation of affected nails; the control group (36 cases, 87 affected nails) was treated with laser only. The patients were treated once a week for 8 weeks and followed up after 6 months. Results The total clinical effective rate (74.67%) of the combined group was higher than that of the control group (63.22%).The clinical effective rate of the three clinical types in the combined group was also higher than that of the control group, especially the proximal subungual type (PSO), but there was no significant difference in statistics. The clinical effective rate of onychomycosis with a score of 9 ≤ SCIO ≤ 15 in the combined group was higher than that of the control group. Conclusion The clinical efficacy of long-pulsed 1064nm laser combined with ketoconazole ointment in the treatment of onychomycosis patients with SCIO value of 9 ≤ SCIO ≤ 15 was significantly higher than that of laser only.

    Clinical efficacy and safety of intravenous injection voriconazole and fluconazole in preventing fungal infection in China: A meta analysis
    LIU Fei-fei, WANG Yan, CHEN Rong, SHEN Jian-zhen
    2019, 14(3):  159-163. 
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    Objective To analyse the clinical efficacy and safety of intravenous injection voricanazole and intravenous injection fluconazole in preventing fungal infection by meta analysis. Method Voricanazole group was the experimental group and fluconazole group was the control group. The randomized controlled trials (RCTs) were retrieved from China National Knowledge Infrastructure (CNKI)、WanFang and VIP Database and references listed in all studies. RCTs meeting inclusive studies were included. The data was extracted, the quality was evaluated and cross-checked by two reviews independently according to Cochrane Handbook for Systematic Reviews of Interventions. Then Meta-analysis were conducted using Stata14.0 software. Results A total of 19 studies and 1492 patients were included. Voriconazole had a better effect and lower adverse events, the difference was statistically significant.(RR=1.20,95%CI=1.14-1.26,P<0.001 AND RR=0.76,95%CI=0.65-0.90,P=0.001,respectively.) Voriconazole also had statistically better effects on the control of infection fever and the clearance of fungi (RR=1.63,95%CI=1.40-1.90,P<0.001 AND RR=1.27,95%CI=1.13-1.44,P<0.001,respectively). Conclusion Meta analysis showed that voriconazole had better effect and prognosis and lower adverse events in preventing fungal infection than fluconazole. voriconazole; fluconazole; fungal infection; randomized controlled trial; meta-analysis

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