Table of Content

28 April 2019, Volume 14 Issue 2

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Original articles

The role of ALS3 and SSA1 gene expression of Candida albicans in the immune mechanism of vaginal candidiasis

GAO Ying, WANG Qiong, LIANG Guan-zhao, SHE Xiao-dong, SHI Dong-mei, SHEN Yong-nian, SU Xiao-hong, LI Dong-mei, LIU Wei-da

2019, 14(2):  65-69. 

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Objective To investigate the effect of ALS3 and SSA1 genes in Candida albicans on immune response of vaginal epithelial cells. Methods Wild strains and ALS3, SSA1 gene knockout strains (SC5314, Δals3, Δssa1) of Candida albicans were cultured and their morphology was determined. Human vaginal epithelial cell line VK2/E6E7 was infected in a certain proportion. The damage of different strains to epithelial cells was evaluated by trypan blue staining and lactate dehydrogenase (LDH) activity detection. The co-culture of inflammatory cytokines and chemokines during infection was evaluated by enzyme-linked immunosorbent assay (ELISA).Results There was no significant difference in the effect of Δals3 on the germ tube length of Candida albicans, while the Δssa1 reduced the germ tube length by about 30%~40% compared with the other two strains (P<0.001). Trypan blue staining and LDH assay showed that the cell damage ability of the three strains was proportional to the fungal load when infected with epithelial cells. Compared with wild strain, the cell damage ability of Δssa1 was significantly reduced when infected with epithelial cells at the same MOI, and the difference was statistically significant (P<0.05), while the impact of Δals3 mutant was weak or even slightly higher. Detection of inflammatory cytokines and chemokines showed that the ability of mutants to induce epithelial cells to produce pro-inflammatory factors and chemokines (GM-CSF, G-CSF, IL-1a, IL-8) was significantly weakened (P<0.05).Conclusion ALS3 and SSA1 gene expression played an important role in the local immune response of vaginal epithelial cells to Candida albicans infection, and SSA1 gene expression was more significant.

Study on the mechanism of NT-89 against Candida albicans based on quantitative proteomics

LIU Yu, YAN Lan, JIANG Yuan-ying

2019, 14(2):  70-77. 

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Objective To analyze the differentially expressed proteins after the treatment of anti-fungal compound NT-89 against Candida albicansby quantitative proteomics iTRAQ technology. Methods Total protein and cell wall proteins of C. albicans were extracted with or without NT-89 treatment. The relative abundanceand differential proteins in protein extracts were detected by iTRAQ. Then the function classification of the differential proteins was performed using GO database. Results 295 differential proteins were detected in extract of total protein (TP), and Ywp1p and Pga10p were most significantly down-regulated in total protein. 6 GPI-anchored proteins in cell wall protein (CWP) extract were found significantly down-regulated. Conclusion NT-89 disrupted the structural integrity and function of cell wall in C. albicans. The iTRAQ technology can provide effective reference information for drug mechanism research.

Clinical value of G test in invasive candidiasis

SHEN Wang, YANG Wen-li, YEL-yan, TAN X-yu, HUANGC-juan, ZHANG Xin

2019, 14(2):  78-82. 

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Objective To explore the diagnostic value of G test in invasive candidiasis (IC) and its relationship with neutrophils, pathogen, and body temperature in patients. Methods Fifty-five patients with IC, 42 patients with invasive aspergillosis(IA), 30 cases of Candida mucosa colonizer, 50 healthy people were collected from September 2015 to June 2018 in Wuyi Traditional Chinese Medicine Hospital Affiliated of Ji nan University, the level of G test were detected by immuneturbidimetry. Diagnostic value of G test in IC patients was assessed by ROC analysis. The relationships between positive rate of G test and neutrophils, pathogen, body temperature in patients with IC were also compared. SPSS19.0 was used for statistical analysis. Results The level of G test in IC patients was significantly higher than these of healthy people, Candida colonizers and IA patients(P<0.05). Among 55 patients with IC, the number of non-Candida albicans (32 cases) was higher than that of Candida albicans (23 cases). Non-IFD group and non-IC group were used as the control group, the area under the ROC curve (AUC) of the G test diagnostic IC patients was 0.899 and 0.782. The positive rate of G test in IC patients with neutrophil normal group was not significant different from those of neutrophil increase group and neutrophil reduction group (P>0.05), but it was significantly higher than that of neutrophil deficiency group (P<0.05).The positive rate of G test in Candida albicans group was not significantly different from that in the Candida tropicalis and Candida glabrata group (P>0.05), but significantly higher than that in other Candida infection group(P<0.05). There were no significant differences in the positive rate of G test between IC patients in different temperature groups(P>0.05). Conclusion G test has high diagnostic value in IC patients. It can distinguish Pathogenic bacteria and colonization bacteria, and has certain value for the identification of IC and IA. The positive rate of G test is related to the neutrophil deficiency and the type of pathogen, and not related to body temperature in IC patients.

Mata analysis of accuracy of clinically isolated fungi by flight mass spectrometry

YU Tian, LIN Fu-gui, HU Li-dong, LI Ke-ke, CHEN Xiao-qing, XIE Yue, SHI Wen-jing, WEI Lian-hua

2019, 14(2):  83-89. 

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Objective To determine the accuracy of clinical isolates based on evidence-based systematic evaluation of flight mass spectrometry and compare it with the accuracy of conventional methods. Methods The main English databases PubMed, Cochrane, Web of Science, and Chinese database CBM, Wanfang, Weipu, and HowNet database were searched for the original literatures of clinical isolates being identified by flight mass spectrometry. Results After screening, 19 articles(6609 strains) were included. The results of meta-analysis showed that the correct rate of lidentification fungi to species was 0.9368 (95% CI=0.9091-0.9598) by flight mass spectrometry. The correct rate of lidentification fungi to species was 0.9104 (95% CI=0.8874-0.9340)by clinical routine methods. Subgroup analysis of the results, including:strain type, research type, sample processing method, gold standard detection range, identification threshold, etc.. Sensitivity analysis showed stable and reliable results.Begg and Egger's results indicated that there was no publication bias in the identification of fungi by flight mass spectrometry, and conventional methods hadidentified a certain publication bias. Conclusion Matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) method is a reliable alternative to clinical routine methods for identifying clinical pathogenic fungi with high accuracy.

In vitro chessboard detection for anti-Cryptococcus effect of rattlerin and fluconazole

LEI Yan, FANG Wei, ZHANG Chao, LIAO Wan-qing

2019, 14(2):  90-94. 

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Objective To determine whether there is synergistic effect of rattlerin combined with fluconazole against Cryptococcus in vitro by the checkerboard method. Methods According to the microdilution method (M27-A3), the fluconazole and rattlerin single drug susceptibility test[FLZ, RAT] and the Fractional Inhibitory Concentration Index (FICI) were used as the judgment indicators of combined drug susceptibility test[FLZ, RAT]. Results The MIC values of rattlerin to 37 strains of Cryptococcus were 32-256 μg/mL. When combined with fluconazole, FICI was ≤ 0.50, which reduced the MIC value of fluconazole by 2-16 times. Conclusion Rattlerin had a certain antifungal effect on Cryptococcus, and it could synergize with fluconazole to improve the sensitivity of fluconazole to Cryptococcus.

Study on the diagnosis of cryptococcal meningoencephalitis by molecular biological techniques

WANG Guang-yu, CHEN Min, HONG Nan, WANG Sheng-qi, LIAO Wan-qing, YANG Ying

2019, 14(2):  95-98. 

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Objective To diagnose the case of meningitis suspected of cryptococcal infection by molecular detection technique. Methods Cerebrospinal fluid samples of patients were collected, DNA was extracted, primers were designed for PCR amplification, and gene detection was performed on the amplified products by DNA chip technology. Results Sample showed positive results for Cryptococcus neoformans. Conclusion PCR amplification of primers designed from ITS conserved sequence and non-culture detection of cerebrospinal fluid samples by DNA chip technology have clinical reference value for deep fungal infections that are difficult to diagnose by traditional methods.

Analysis of 1031 cases of dermatophytosis and the pathogenic dermatophytes

JIANG Wei-wei, HU Dong-ying, HOU Qing, DU Ming-wei, LI Hang, DENG Yu-chen, LI Shuai, CAO Yun, ZHENG Fang-wei, LING Li-yan, YU Xiao-tian, FANG Wen-jie, ZHAO Jin, PAN Wei-hua, LIAO Wan-qing

2019, 14(2):  99-103. 

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Objective To understand the clinical characteristics, susceptible factors, distribution of fungal pathogens of dermatophytosis in Shanghai. Methods All 1031 cases of dermatophytosis presented with typical clinical features and positive culture results were collected during January 2013 to December 2017 in our outpatient department for analysis. SPSS 23.0 was used in statistic analysis. Results Of 1031 cases of dermatophytosis, 535 were males and 496 were females, 1033 strains of dermatophytosis were isolated. The peak incidence of dermatophytosis was 21-30 years old and 31-40 years old. 71 cases (6.89%) had one or more other fungi isolated from patients with dermatophytosis, and 2 cases (0.19%) had two kinds of dermatophytes, in which the most isolated other fungi were Rhodotorula. The most common dermatophytosis was onychomycosis (n=344,33.37%), followed by tinea cruris (n=200,19.40%), tinea corporis (n=162,15.71%), tinea pedis (n=138,13.39%), tinea manus (n=108,10.48%), tinea facialis (n=45,4.36%) and tinea capitis (n=34, 3.30%), in which there were significant gender differences in tinea cruris and onychomycosis. In the distribution of strains, the top three were Trichophyton mentagrophytes (n=542, 52.47%), Trichophyton rubrum (n =397, 38.43%) and Microsporum canis (n=66, 6.39%), in which there were significant gender differences between Trichophyton rubrum and Microsporum canis. Conclusion The main type of dermatophytosis in our hospital was onychomycosis, and the main pathogen was Trichophyton mentagrophytes. The distribution and epidemic trend of dermatophytosis and dermatophytosis are similar to those in most areas of China in recent years, but it has its own characteristics.