Table of Content

28 April 2022, Volume 17 Issue 2

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Original articles

Transcriptome study of mouse bone marrow-derived macrophages infected by Aspergillus fumigatus spores

XU Fengjiao, LOU Xinyu, WANG Yuanzhou, HAN Ziyu, ZHONG Guowei

2022, 17(2):  94-98. 

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Objective To reveal the important genes related to phagocytosis, immune responses, and clearance of spores in the mouse bone marrow-derived macrophages (BMMs) infected by Aspergillus fumigatus. Methods Mouse primary bone marrow cells were extracted, induced to differentiate into macrophages, and challenged with A1160^c conidia. After infection, the total RNAs of BMMs in the infected group and the control group were extracted and analyzed by RNA-sequencing. Results The primary mouse bone marrow cells were successfully induced to differentiate into BMMs. Compared to untreated control BMMs, the transcription of Toll-like receptor (TLR) Tlr2 and C-type lectin receptors Clec4e and Clec5a were upregulated in conidia-infected BMMs. The NF-κB pathway was also activated through increasing the transcription levels of inflammation related genes such as Il1b, Tnf, Nfkb1, Tnfsf14, Jun, Gsk3b, Tec and Cd40, etc. Meanwhile, the expression of chemokines such as Cxcl2, Cxcl10 and Ccl3 also increased. In addition, the transcription levels of Bax, Nos2, Glul, and Nlrp3, which were related to the generation of reactive oxygen species (ROS), were obviously upregulated. Conclusion To adapt infection of A. fumigatus spores, BMMs activated TLR and C-type lectin receptors, NF-κB pathway, and other immune pathways, exhibiting an elevated immune response and spore clearance potential.

Clinical and laboratory analysis of bloodstream infection caused by Candida tropicalis

CHENG Wei, DENG Jing, LIU Ya, SHU Ling, CHEN Zhixing, DAI Zhongqiu, KANG Mei

2022, 17(2):  99-102,114. 

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Objective To analyze the clinical characteristics and laboratory results of bloodstream infection caused by Candida tropicalis in order to provide reference for clinical and prevention. Methods The clinical data of patients with bloodstream infection caused by Candida tropicalis in West China Hospital Sichuan University from January 2018 to December 2020 were collected, and the related clinical characteristics and laboratory examination of the patients were analyzed retrospectively. Results The 27 patients with bloodstream infection caused by Candida tropicalis had basic diseases and two or more risk factors. (1,3)-β-D-glucan test had a good sensitivity in the auxiliary diagnosis of Candida tropicalis bloodstream infection. Other inflammatory indexes such as high sensitive C-reactive protein, procalcitonin and interleukin-6 were increased in varying degrees.The sensitivity rates of 27 strains of Candida tropicalis to azole antifungal drugs were itraconazole(77.8%),voriconazole(48.2%), fluconazole(48.2%), posaconazole(37.0%). The sensitivity rate to amphotericin B, caspofungin and micafungin was 100%. Conclusion Candida tropicalis bloodstream infection often occured in patient with underlying diseases and a variety of risk factors Candida tropicalis had poor sensitivity to azole drugs, so we should pay attention to drug sensitivity results in clinical treatment.

Using Galleria mellonella model to investigate the role of SMT3 gene in regulating pathogenicity of Candida albicans

HU Jing, LI Aihong, FENG Jinrong

2022, 17(2):  103-108. 

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Objective To investigate the role of SMT3 gene in regulating pathogenicity of Candida albicans. Methods To delete the SMT3 gene, a transient CRISPR/Cas9 system was used. The genotype of the SMT3 deletion strain was checked by colony PCR. Invasive growth and biofilm formation of the SMT3 deletion strain were studied. To check the pathogenicity of C. albicans cells, wild-type and the SMT3 deletion strain were injected into the larvae of Galleria mellonella. The survival rate of infected larvae was checked daily and analyzed. As well, the morphology of C. albicans in the larvae was checked using a PAS staining. Moreover, the damaging effect of the SMT3 deletion strain on the hemolymph of G. mellonella was investigated. Results The SMT3 homozygous deletion strain was successfully constructed. Deletion of SMT3 gene significantly increased the invasive growth on solid media, and also elevated the ability of biofilm formation. Deleting SMT3 decreased the survival of infected G. mellonella, An average survival of 3.5 days in this group is significantly lower than the wild type group (8.5 days). In addition, deleting SMT3 caused a significant decrease in total hemolymph of infected larvae, (1.4×10⁷ cells per mL). Conclusion SMT3 gene plays important roles in regulating pathogenicity of C. albicans in G. mellonella model.

Analysis of 2 cases of mucormycosis and related literature

MA Shujun, ZHANG Furong, TAO Sujiang, YU Yongtao, WANG Ying

2022, 17(2):  109-114. 

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Objective To improve the understanding of mucormycosis of nostril-brain type. Methods The diagnosis, treatment and prognosis of 2 cases of rhinocerebral mucormycosis secondary to hematologic diseases which we assisted the treatments were analyzed. Relevant case literatures were also reviewed. Results All 2 cases were confirmed to be in remission stage of hematologic disease. Amphotericin B intravenous drip and external wet compress were used. Skin infection was controlled in 2 cases, but only 1 case was successfully treated. A review of 61 cases of rhinocerebral mucormycosis showed that 52 patients had been treated with amphotericin B and surgery and the mortality rate of this group was 55.8%. The high risk factors included uncontrolled diabetes and hematopathy or solid organ transplantation. Conclusion The mortality rate of hematologic diseases combined with mucormycosis was high. Early diagnosis, effective treatment and appropriate operation timing can improve the survival rate of patients.

Application of polymerase chain reaction in the diagnosis of tinea incognito with fungal blisters or pustules

XIA Hui, YANG Hong, ZHANG Pengfei, MO Xiaohui, TAN Fei

2022, 17(2):  115-119,128. 

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Objective To analyze the clinical value of polymerase chain reaction (PCR) in fungal blisters or pustules to diagnose tinea incognito. Methods From July 2020 to October 2021, thirty-four patients presenting tinea incognito with blisters or pustules in the outpatient department of our hospital (Shanghai Skin Disease Hospital) were included. PCR was performed in blister or pustule fluid specimens from 34 patients and compared by fluorescent staining microscopy and fungal culture. Results Blister or pustule fluid specimens were collected from a total of 34 patients. For fluorescent staining microscopy, fungal culture and PCR, the sensitivities were 100%, 77.8% and 100%, the specificities were 87.5%, 100% and 75%, the positive predictive values were 85.7%, 100% and 75%, the negative predictive values were 100%, 80% and 100%, the accuracy were 94.1%,88.2% and 88.2%, respectively. DNA extraction from blister or pustule fluid, PCR amplification and sequencing comparisons were completed within 12 hours, which identified six species and 22 cases of fungi (one case was identified to genus). Conclusion In the diagnosis of tinea incognito with fungal blisters or pustules, PCR has good sensitivity and accuracy. PCR can determine the fungal species compared with fluorescent staining microscopy and shorten the time compared to fungal culture, which can provide reference value for rapid clinical diagnosis and timely treatment of tinea incognito.

Investigation of superficial mycoses in military academy cadets

TANG Renjie, WANG Ruina, WANG Zhi, LIU Haiyan, LIU Xu, LI Honglei, LIU Jiacun, ZHANG Dazhi, YAN Lan

2022, 17(2):  120-123. 

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Objectives To investigate the incidence and treatment of superficial mycoses in cadets of a military academy. Methods A questionnaire survey was carried out during the physical examination. The cases of superficial mycoses were further examined by microscopy. Results A total of 1733 cadets were examined clinically and submitted the questionnaire. There were 186 (10.73%) cadets diagnosed with superficial mycoses after entering the military academy. The incidence of tinea pedis (n=147) was higher than any other superficial fungal infections. The prevalence rate of superficial mycoses in summer and autumn was significantly higher than that in winter and spring (P<0.001). Statistic analysis demonstrated that the incidence of dermatophytosis was significantly different in different period of military service(P<0.001). In this survey, 70.87% of patients took topical antifungal drugs, while 6.80% received the combination of topical and systematic therapy. The azoles were highest administered. It was 50.62% on patients' satisfaction of treatment with antifungal agents. The relapse rate of superficial fungal infections was 51.46%. Conclusions The incidence of superficial mycoses was high in cadets in a military academy. Attention should be paid to enhance the health publicity and education and to promote the rational use of drugs for cadets in military academies.

Study on antifungal activity of nicotinamide and the synergistic effect of nicotinamide to itraconazole against Aspergillus fumigatus in vitro

CHEN Xianzhen, ZHU Xinlin, JANG Weiwei, HU Dongying, ZHANG Keming, FANG Wenjie, PAN Weihua, LIU Xiaogang, LIAO Wanqing

2022, 17(2):  124-128. 

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Objective To investigate the effect of nicotinamide (NAE) alone or in combination with itraconazole (ITR) against Aspergillus fumigatus. Methods We investigates the antifungal effects of ITR/NAE alone and in combination against 15 isolates of Aspergillus fumigatus according to the clinical and laboratory standards institute (CLSI) M38-A2. The minimum inhibitory concentrations (MICs) and the fractional inhibitory concentration index (FICI) were used to classify drug interactions. Real-time PCR was used to test the relative expression of gene in Aspergillus fumigatus treated with nicotinamide/itraconazole. Results Niacinamide had inhibitory effect on Aspergillus fumigatus, the MICs was 12.8-25.6 mg/mL. The FICI of the drugs in combination was less than 0.5, revealing synergism between NAE and ITR. The expression of membrane protein genes and efflux pump related genes was down-regulated under nicotinamide/itraconazole. Conclusions Nicotinamide inhibited Aspergillus fumigatus and enhanced itraconazole against Aspergillus fumigatus. This effect was related to the decrease of membrane protein and efflux pump protein gene expression of Aspergillus fumigatus.

Analysis of 2181 cases of superficial mycosis caused by Trichophyton rubrum in Guangzhou region

XIE Zhenmou, HUANG Jinmei, XUE Yaohua, LIU Hongfang, FANG Mingheng, ZENG Weiying

2022, 17(2):  129-131,151. 

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Objective To understand the clinical characteristics of superficial mycoses caused by Trichophyton rubrum in Guangzhou. Methods The clinical data of patients who came to Dermatology Hospital, Southern Medical University from January 2012 to June 2020 for fungal culture examination and identified Trichophyton rubrum were analyzed. Results In this analysis, 2181 cases of superficial mycosis caused by Trichophyton rubrum were enrolledincluding 1407 cases of onychomycosis (64.51%), 324 cases of tinea pedis (14.85%), 249 cases of tinea cruris (11.42%), 144 cases of tinea corporis (6.60%), 37 cases of tinea manus (1.70%), 20 cases of tinea capitis (0.92%). There was statisticaI significance in the constituent ratio of tinea corporis, tinea cruris and onychomycosis between different gender. Different ages of superficial mycosis distribution had obvious difference. Conclusion The superficial mycoses caused by Trichophyton rubrum in Guangzhou was similar to the superficial mycoses caused by various strains reported in China, but the proportion of onychomycosis was higher.

Antifungal drug susceptibility of Fonsecaea spp.

WANG Huiwei, JIANG Weiwei, XU yuan, LI Yingfang, LI Hang, ZHU Xinlin, CHEN Tianyang, Wang Qilong, CHEN Min, PAN Weihua, LIAO Wanqing

2022, 17(2):  132-136. 

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Objective To study Fonsecaea spp. strains isolated from China to understand the characteristics of antifungal drug susceptibility and to provide data basis for the development of precise antifungal drug regimens for dark fungal diseases caused by Fonsecaea spp. infections in China. Methods Using the broth microdilution method (CLSI M38-A2 method) recommended by the American Clinical Laboratory Standardization Committee for filamentous fungi, in this study, in vitro drug sensitivity tests were conducted on 32 clinical isolates of Fonsecaea spp. against 11 clinical antifungal drugs. We also systematically analyzed the drug sensitivity characteristics and clinical prognosis of 83 reported cases of Fonsecaea spp. infection in China and abroad. Results Fonsecaea spp. was most sensitive to the antifungal activity of second-generation azole antifungal drugs, including:posaconazole (MIC₅₀=0.25 μg/mL, MIC₉₀=0.5 μg/mL), esaconazole (MIC₅₀=0.25 μg/mL, MIC₉₀=0.5 μg/mL), voriconazole (MIC₅₀=0.125 μg/mL, MIC₉₀=0.25 μg/mL). While amphotericin B (MIC₅₀=4 μg/mL, MIC₉₀=8 μg/mL), fluconazole (MIC₅₀=8 μg/mL, MIC₉₀>8 μg/mL), and echinocandins (MEC₅₀=8 μg/mL, MEC₉₀>8 μg/mL) had lower antifungal activity against Fonsecaea spp. Conclusion For the antifungal treatment of Fonsecaea spp. infection, it is advisable to give preference to second-generation azoles (posaconazole, esaconazole, voriconazole, etc.). The in vitro combined drug sensitivity profile of clinical isolates of Fonsecaea spp. warrants further study.

The effects of calcineurin on asexual development, polarity and stress responses in Talaromyces marneffei

HUANG Qihua, LUO Hong, PAN Kaisu, LIAO Liuwei, ZHENG Yanqing, CAO Cunwei

2022, 17(2):  137-145. 

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Objective To determine the functions of calcineurin B (cnaB) and meanwhile further investigate the effects of calcineurin A (cnaA) from Talaromyces marneffei on asexual development, polarity and stress responses. Methods The growth, morphogenesis and external stress responses of strains deleted for cnaB (ΔcnaB strain) or CnaA (ΔcnaA strain) were observed and compared with those of parental strain FRR2161 (wild-type) at both 25℃ and 37℃. Results Both ΔcnaB and ΔcnaA gained severe defects in asexual development, polarity and stress responses. ΔcnaA strain exhibited more severe defects in cellular lysis, separation and resistance to diverse stressors. Conclusion Both cnaB and cnaA exert a key role on Talaromyces marneffei asexual development, polarity and stress responses, but cnaA seems to play a major role in some biological procedures like cellular separation, oxidative stress and salt stress.

Diagnostic value of serum and bronchoalveolar lavage fluid galactomannan test in non-neutropenic patients with different types of pulmonary aspergillosis

ZHANG Chuanming, CAO Hongyu, SUN Jide, NIU Siqiang, WANG Zhu, PU Shuli, XU Xiuyu

2022, 17(2):  146-151. 

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Objective To investigate the diagnostic value of serum and bronchoalveolar lavage fluid (BALF) galactomannan (GM) test in different types of pulmonary aspergillosis, and to determine the best cut-off value of BALF GM test for different pulmonary aspergillosis. Methods From January 2017 to March 2021, clinical data of 146 non-neutropenic patients diagnosed with pulmonary aspergillosis who underwent serum and BALF GM test in the First Affiliated Hospital of Chongqing Medical University were collected, including 97 cases of IPA, 39 cases of CPA and 10 cases of ABPA. 132 patients with other lung diseases in the same period were randomly selected as the control group. The diagnostic performance of serum and BALF GM test for different types of pulmonary aspergillosis was analyzed. Results In non-neutropenic patients, the best cut-off values of BALF GM test for IPA and CPA were 1.0 and 0.91, the sensitivity were 74.2% and 74.4%, and the specificity were 83.3% and 81.1%, respectively. However, the diagnostic value of BALF GM test for ABPA was limited (AUC=0.648). At a cut-off value of 0.5, the diagnostic sensitivity of serum GM test for IPA and CPA was 41.2% and 10.3%, respectively, and the diagnostic value for ABPA was even worse (AUC<0.5). Conclusion In non-neutropenic patients, BALF GM test has a good diagnostic effect on IPA and CPA, but its diagnostic effect on ABPA is limited. The diagnostic sensitivity of serum GM test to IPA and CPA is low, and its diagnostic value of ABPA is worse.

Majocchi granuloma of head and face caused by Trichophyton rubrum

WU Ling, GU Lihong, CHEN Yuchong

2022, 17(2):  155-157. 

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We reported a case of Majocchi granuloma with inflammation around hair follicles in the scalp caused by Trichophyton rubrum, which was diagnosed by histopathology and fungal culture and strain identification. The patient was a 84-year-old male with papules and nodules on the bilateral temporal scalp for more than 1 year. Multiple red papules and nodules were observed on the bilateral temporal scalp, and some of them are fused into pieces. Histopathological examination showed an inflammatory infiltration area in the middle and lower dermis, with predominantly neutrophils and many tissue cells and multinucleated giant cells. Many hyphae and spores were observed in a hair shaft. Trichophyton rubrum was identified by fungal culture and strain identification. This patient was finally diagnosed as Majocchi granuloma on the scalp caused by Trichophyton rubrum. This patient was treated with itraconazole 0.2 g, twice a day, orally for 2 months. The patient's rash subsided. He is currently under follow-up.