Chinese Journal of Mycology

The role of EGFR in the pathogenesis of vaginal candidiasis

ZHANG Jing-yun, PENG Jing-wen, WANG Qiong, GAO Ying, CHEN Wan-xin, SHEN Yong-nian, LI Dong-mei, SHE Xiao-dong, LIU Wei-da

2020, 15(2): 65-71.

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Objective To investigate the role and molecular mechanism of EGFR in the pathogenesis of vaginal candidiasis. Methods Vaginal epithelial cells (VK2/E6E7 cells) were cultured. After stimulation with Candida albicans, the expression of immune factors in EGFR and possible related immune pathway factors were detected by real-time PCR. The EGFR-siRNA VK2/E6E7 cell model was constructed and co-cultured with Candida albicans. The cytokines secreted by vaginal epithelial cells and the protective ability against Candida albicans infection were detected by ELLISA and automatic cell detector. A mouse model of vaginal Candida infection was constructed. qPCR was used to detect the expression of EGFR and immune pathway factors in vaginal tissues. The changes of fungal load and inflammatory cells in vaginal tissues were detected after EGFR phosphorylation inhibitor blocking pathway. Results qPCR assay showed that EGFR, STAT3, GM-CSF and IL-1β were up-regulated in VK2/E6E7 cells infected with Candida albicans, and were statistically significant (P <0.05). ELLISA assay showed the expressions of GM-CSF, IL-8, IL-1β and MIP-3α were significantly decreased in EGFR-siRNA VK2/E6E7 cells infected with Candida albicans (P<0.05). The results of automatic cell detector showed that the defense capacity and cell activity of EGFR knockout cells after 30 hours' infection of Candida albicans were significantly lower than normal cells. The results of qPCR showed that the expression of EGFR, HER2, STAT3 and IL-8 following the vaginal infection of Candida albicans in the mouse was statistically significant (P<0.05); after local application of phosphorylation inhibitor, the lavage fluid load in the mouse vagina was significantly higher than that of the control group with EGFR pathway being blocked and there was a statistical difference on the 7th day after infection (P<0.05). Conclusion The EGFR and its pathway factors should play an important role in the pathogenesis of vaginal candidiasis.

Study on the antifungal effect of Aspergillus fumigatus, Trichophyton mentogrophtes and Cryptococcus neoformans to alkaloids from Sophora alopecuroides

ZHANG Juan, LI Xiao-hui, Feng Xue-ling, ZHANG Jing-jing

2020, 15(2): 72-77.

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Objective To compare the susceptibility of Aspergillus fumigatus, Trichophyton mentogrophtes and Cryptococcus neoformans cultured with Sophora alopecuroides total alkaloids, sophocarpine, matrine, sophoridine and oxymatrine. The results will lay a good foundation for further development and utilization of alkaloids from Sophora alopecuroides. Methods Antifungal activity of five kinds of alkaloids against Aspergillus fumigatus, Trichophyton mentogrophtes and Cryptococcus neoformans were detected with both standard broth microdilution method and cylinder-plate assay. Results Sophora alopecuroides total alkaloids, sophocarpine, matrine, sophoridine and oxymatrine were active against Aspergillus fumigatus, Trichophyton mentogrophtes,Cryptococcus neoformans by cylinder-plate method. The MFCs of Sophora alopecuroides total alkaloids, sophocarpine, matrine and sophoridine for Aspergillus fumigatus reached 25.0, 4.25, 5.0, 3.125 mg·mL^-1 respectively. The MFC of oxymatrine for Aspergillus fumigatus was over 500mg·mL^-1.The MFCs of the five types of alkaloids for Trichophyton mentogrophtes were 3.125,2.125,5.0,1.5625,125 mg·mL^-1; the MFCs of Cryptococcus neoformans were 3.125,4.25,10,6.25,1000mg·mL^-1 respectively. Conclusion Sophora alopecuroides total alkaloids, sophocarpine, matrine and sophoridine showed good antifungal activities. But the inhibitory effects of oxymatrine against Aspergillus fumigatus, Trichophyton mentogrophtes and Cryptococcus neoformans were less than that of the other alkaloids.

Analysis of distribution and drug resistance of clinical Candida strains in 497 cases

WANG Jun-ting, LIU Yong

2020, 15(2): 78-82.

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Objective To explore the type, distribution and drug resistance of inpatient's Candida infection in Shenyang, so as to provide theoretical basis for clinical prevention and rational drug use of Candida infection. Methods The clinical data of Candida strains isolated from inpatients in Sheng Jing Hospital affiliated to China Medical University from 2015 to 2018 were collected retrospectively, and the data and results of strain identification and drug sensitivity test were statistically analyzed. Results A total of 497 strains of Candida were detected, of which the main strains were Candida parapsilosis (175, 35.2%), followed by Candida albicans (127, 25.6%), Candida pelliculosa (51, 10.3%), Candida glabrata (42, 8.5%) and Candida tropicalis (36, 7.2%). The distribution of the strains was statistically different (P=0.004). The hosts were mainly of patients aged 0-9 and 50-69 years old. In 497 strains of Candida, the isolates of from blood was accounted for 81.5%,followed by those from hydrothorax or ascitic fluid (4.2%), drainage fluid (4.2%) cerebrospinal fluid (1.4%) and urine were (1.4%). There was significant difference in the distribution of strains among different types of samples (P=0.003). The resistance rates of clinical common Candida to fluconazole, voriconazole, flucytosine, itraconazole, and amphotericin B were 3.0%, 2.2%, 4.0%, 5.0% and 4.0%, respectively. Conclusion The infection of Candida albicans in hospital was the most common. In the treatment of newborns, elderly patients and patients with digestive system diseases, we should be alert to the occurrence of double infection of Candida.

Analysis of strain distribution and drug resistance of Candida vaginal isolates in women from a region of Guizhou Province

ZHAO De-jun, YANG Lin

2020, 15(2): 83-87.

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Objective To investigate the distribution characteristics and drug resistance of Candida isolates from vaginal secretion specimens in women from a region of Guizhou Province, and to provide a reference for clinical treatment. Methods A total of 2055 female cases in a region of Guizhou Province from January to December 2018 were collected. Culture and drug sensitivity test of Candida vaginal isolates from secretion samples, and statistical analysis of the results were performed. Results A total of 543 Candida strains were isolated and identified, with the highest isolation rate of Candida albicans, accounting for 40.5%,Candida glabrata and Candida Parapsilosisis accounting for 35.0% and 19.7%, respectively. Infected patients were most common in the age group of 26~30 (39.2%), followed by 21~25 age group (25.4%) and 31~35 age group (19.2%); 90.4% of infected patients were 21 to 40 years old. The resistance of different Candida antifungal drugs was quite different, and no resistant Candida isolate to micafungin, nystatin and caspofungin was found. Non-Candida albicans isolates were significantly more resistant to itraconazole, fluconazole and ketoconazole(P<0.01), there was no significant difference in the rates of 5-fluorocytosine resistance(P>0.05). Conclusion Candida albicans is more common isolate among women in this area of Guizhou Province, and the drug resistance of different Candida species was not similar. Clinical reasonable treatment plan should be developed according to different pathogenic resistant characteristics.

The inhibitory effects of natural antibacterial agent on 5 pathogenic microorganisms

LIU Guang-rong, ZHAI Chun-tao, JIN Min-rong, LI Cheng-liang

2020, 15(2): 88-92.

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Objective To study the inhibitory effects of natural antibacterial agent on Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Candida albicans and Aspergillus niger. Methods The diameter of inhibition zoon and the minimum inhibitory concentration (MIC) of natural antibacterial agent against 5 pathogenic microorganisms and MICs of 10 strains of Staphylococcus aureus were determined by cup and dish method,microporous plate method. Result The diameter of inhibition zoon of 1% natural antibacterial agent against S.aureus, E.coli, P.aeruginosa, C.albicans and A.niger were 14.47mm, 10.67mm, 9.13mm, 28.13mm and 10.53mm, respectively. The MICs of natural antibacterial agents against S.aureus, E.coli, P.aeruginosa, C.albicans and A.nigerwere 0.08%, 0.16%, 0.63%, 0.08% and 0.16%, respectively. The MICs of Sensiva SC50 against these five microorganisms were 0.16%, 0.16%, 0.31%, 0.16% and 0.13%, respectively. The MICs of 10 strains of S. aureus were 0.08%、0.06%、0.05%、0.09%、0.06%、0.07%、0.10%、0.06%、0.09%、0.07%, respectively. Conclusion The natural antibacterial agent had a certain inhibitory effect on the five microorganisms, the inhibitory effects on S. aureus and C.albicans were better than the positive control, the inhibitory effects on E.coli, and A.niger were similar to the positive control, and the effect on P.aeruginosa was slightly worse than the positive control. And all the 10 stains of S. aureus showed good inhibitory effect.

In vitro antifungal susceptibility of polyhexamethylene biguanide against common superficial fungal species

TAN Jing-wen, XU Hong, SONG Jin-feng, GAO Zhi-qin, YANG Hong, YANG Lian-juan, WEN Hai

2020, 15(2): 93-96.

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Objective To determine the in vitro antifungal susceptibility of polyhexamethylene biguanide (PHMB) against common superficial fungal species. Methods We used the standard broth microdilution method and agar dilution method to evaluate the minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC) of PHMB against the clinical isolates of Trichophyton rubrum, T. mentagrophytes, Microsporum canis and Candida albicans. Results Broth microdilution method showed that MIC range of PHMB powder against T. rubrum was 1.0~4.0 μg/mL, the MFC range was 2.0~8.0 μg/mL. The MIC range of T. mentagrophytes was 2.0~4.0 μg/mL and the MFC was 4.0 μg/mL. To M. canis the MIC range was 2.0~4.0 μg/mL and MFC range was 2.0~4.0 μg/mL. To C. albicans the MIC range turned out to be 0.3~5.0 μg/mL and the MFC range was 5.0~10.0 μg/mL. The agar dilution method showed there were none T. rubrum, T. mentagrophytes and M.canis colony growth on agar plate when PHMB liquid at the concentration of 1.25%. There was none C. albicans colony growth at the concentration of 0.32%. After added the penetration enhancer of azone, the result showed no difference. Conclusions PHMB showed great in vitro efficacy against T. rubrum, T. mentagrophytes, M. canis and C. albicans, suggesting a potential therapeutic choice for treatment of superficial fungal infection.

Study on mucocutaneous candidiasis in infants: the clinical characteristics and in vitro susceptibility of pathogenic yeasts

KAN Si-yue, CAI Qing, JING Yun, CHEN Wei, HU Su-quan, YANG Lian-juan, LIU Wei-da

2020, 15(2): 97-100.

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Objective To analyze and investigate the clinical characteristics of mucocutaneous candidiasis in infants, and to determine in vitro antifungal susceptibility of pathogenic yeasts. Method 105 patients with clinical and mycological diagnosis of mucocutaneous candidiasis, aged from 0~3 years old, were enrolled. Meanwhile, the questionnaires of related data from patients were collected and analyzed. The strains were identified by CHROMagar medium and API20AU test card. Antifungal susceptibility testing was performed according to CLSI documents M27-A3, using 5 drugs including miconazole, econazole, ketoconazole, bifonazole and terbinafine. Results The subjects were between 0.4 and 36 months old (average age, 10.67 months old), and most of them were within one year old (75%). Skin lesions were mainly found in the diaper area, followed by neck and armpit. 189 strains were isolated from those patients; 81.5% were C.albicans, 10.6% were C.parapsilosis, 3.2% were C.glabrata. Miconazole and ketoconazole showed the lowest MIC₉₀ (0.25 μg/mL) against C.albicans. The MIC₉₀ of Ketoconazole and econazole among all strains including non-albicans Candida were 0.25 μg/mL and 2 μg/mL respectively. Bifonazole and terbinafine showed the highest MIC₉₀ (16 μg/mL) against C.albicans, and the MIC₉₀ among non-albicans Candida were 8 μg/mL and 16 μg/mL respectively. Conclusion Mucocutaneous candidiasis was more frequent in infants within one year old. The lesions mainly located in the warm and humid areas such as the diaper area, neck and armpit. C.albicans was still the predominant species of mucocutaneous candidiasis in infants, while C.parapsilosis was the main species in non-albicans Candida. Miconazole and ketoconazole, especially ketoconazole, were better external drug s for the treatment of mucocutaneous candidiasis in infants.

Retrospective analysis of chromoblastomycosis in mainland China: a review of 52 cases

SUN Qi-lin, YU Min, CHEN Jun, LIU Yang, LIAO Wan-qing, XU Hui, YANG Ya-li

2020, 15(2): 101-105.

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Objective To summarize and explore the epidemiology, diagnosis and treatment of chromoblastomycosis in mainland China. Methods The relevant literatures about chromoblastomycosis in mainland China were collected from the electrical databases since 1994, such as Chongqing Weipu, China knowledge Network, Wanfang Database and PubMed database. Results A total of 29 literatures and 52 cases were searched. The average age of onset was 53.9 years old, and the ratio of male to female was 5.5:1; the disease was common in farmers and workers, with a history of trauma in the distal part of the extremities; most of the early onset is mainly papules, followed by plaques, nodules and ulcers; mycological examination strains is mainly Fonsecaea pedrosoi, and the most common treatments are itraconazole and terbinafine. The prognosis is good. Conclusions Chromomycosis is a chronic and refractory subcutaneous granulomatous disease with various clinical manifestations. It can cause teratogenic disability and even cancer in the late stage. Early diagnosis and treatment of the disease is very important.

The role of oxidative stress in the treatment of onychomycosis with the 1064-nm long pulse Nd: YAG laser

XING Zhao-jing, CAI Hong, LIU Wei

2020, 15(2): 106-110.

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Objective To investigate the inhibition/killing effect of 1064-nm long pulse Nd:YAG laser on Trichophyton rubrum in vitro-the most common pathogen causing onychomycosis-and the role of oxidative stress mediated by reactive oxygen species (ROS). Methods Colonies of Trichophyton rubrum were irradiated by using 1064-nm long pulse Nd:YAG laser with various pulse numbers (7 pulses, 9 pulses, 11 pulses and 13 pulses) and other parameters were fixed. Colony areas were photographed to draw growth curves before treatment and 1, 3, 5 and 7 days after treatment and assess the minimum number of pulses with the effect of growth inhibition. The colony and spore suspension of Trichophyton rubrum were irradiated with this pulse number to compare the changes of colony morphological structure, the relative activity of the spore suspension by WST-8, and the expression level of ROS after different treatments by using DCFH-DA probe. Results Nine pulses were the minimum numbers to induce the inhibitory effect. With the increase of energy, the inhibition/killing effect of laser on Trichophyton rubrum was enhanced, while the relative vitality decreased with the increase of time, showing the laser intervention had energy and time dependence. After laser intervention, mycelia lost their normal morphological structure, and the fluorescence intensity and ROS content of the spore suspension were higher than those of the unirradiated, P=0.0157 <0.05. Conclusion The 1064-nm long pulse Nd:YAG laser could inhibit/kill Trichophyton rubrum. The oxidative stress mediated by ROS may play an important role in the treatment of onychomycosis inducing by Trichophyton rubrum with 1064-nm long pulse Nd:YAG laser.

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