Table of Content

28 June 2020, Volume 15 Issue 3

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Original articles

Study on the expression of dual specificity phosphatase in alveolar epithelial cells during Aspergillus fumigatus infection

ZHAO Rui, CHEN Fang-yan, HAN Li

2020, 15(3):  129-133. 

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Objective To study the expression of dual specificity phosphatases (DUSPs) in alveolar epithelial cells in response to Aspergillus fumigatus infection and the release of inflammatory factors. Methods During Aspergillus fumigatus infection, ELISA method was used to detect the release of inflammatory factors in the supernatant of alveolar epithelial cells, western blot was used to detect the expression of DUSP2, DUSP6 and DUSP8 and the expression and phosphorylation level of p65, ERK1/2 and p38 in alveolar epithelial cells. Results During the infection of alveolar epithelial cells by Aspergillus fumigatus B5233 conidia, the content of TNF-α, MCP-1, IL-12p40, IL-1β, IL-6 and IL-8 in the cell supernatant gradually increased significantly. And the expression of intracellular DUSP2 gradually increased, the expression of DUSP6 gradually decreased, but the expression of DUSP8 did not change significantly. The phosphorylation level of ERK1/2 and p65 increased significantly. Conclusion In response to Aspergillus fumigatus infection, the expression of DUSP2 increased and the expression of DUSP6 was inhibited in alveolar epithelial cells, but there was no change about DUSP8 expression. At the same time, intracellular NF-κB signal and MAPK signal were activated to release a variety of inflammatory factors.

Construction and identification of yeast two-hybrid cDNA library of Cryptococcus neoformans

ZHAO Jing-yu, LIU Yi-shu, FANG Wei, LIAO Wan-qing, WANG Gui-zhen, GU Ju-lin

2020, 15(3):  134-137. 

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Objective To lay foundation for investigating interaction mechanism between Cryptococcus neoformans and host cells, yeast two-hybrid cDNA library of H99 cells was constructed. Methods mRNA was isolated from Cryptococcus neoformans (H99) in the mid-log phase. cDNA were then synthesized using biotin-conjugated Oligo (dT) primer in the 5’end. The double-strand cDNA was ligated to adapter and passed the cDNA size fractionation columns. The primary cDNA library was constructed by BP recombination and the cDNA expression library was constructed by LR recombination. Results The primary library contained a total clones of 1.13×10⁷ CFU with a recombination rate of 90.63%. The constructed cDNA library contained total clones of 1.25×10⁷ CFU, and the recombination rate was 100%. Conclusion A high-quality cDNA library of Cryptococcus neoformans was constructed successfully. These data might be useful for screening the host proteins interacting with Cryptococcus neoformans and for following functional studies of Cryptococcus neoformans in the future.

The effects of calcineurin A on the pathogenicity of Talaromyces marneffei

HUANG Chun-yang, PAN Kai-su, LI Xiu-ying, CAO Cun-wei, ZHENG Yan-qing

2020, 15(3):  138-144. 

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Objective To investigate the effects of calcineurin A from T. marneffei on pathogenicity. Methods The morphogenesis, cell wall integrity, external stress responses, growth of co-cultured with macrophages, and constructed animal infection model were observed to determine the roles of the CnaA on the pathogenicity of T. marneffei. Results Deletion of the CnaA gene in T. marneffei resulted in a strain with significant defects in morphogenesis, cell wall integrity, and resistance to various stressors. The conidia of ΔcnaA strain co-incubated with RAW264.7 macrophages were rapidly phagocytosed. By 24 h, the ΔcnaA strain showed much slower growth than the wild-type cells. At 48 h, very few ΔcnaA strain remaining in macrophagesshowed that most of the ΔcnaA strain had been killed and degraded. In the infected mouse model, the mortality and tissue fungi burden of ΔcnaA mutant infected mice were significantly lower than those of wild-type. Conclusion CnaA of T. marneffei plays a key role in controlling fungal morphogenesis and the response to internal and external stresses, as well as in the host immune response and fungal pathogenicity.

In vitro antifungal susceptibility of 10 Chinese herbal monomers against Sporothrix globosa

QU Xin-yao, LI Shan-shan, QU Yue, Abdulhakim Salad Nageye, SONG Yang, CUI Yan

2020, 15(3):  145-149. 

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Objective To investigate the antifungal effects of 10 Chinese herbal monomers against Sporothrix globosa in vitro. Methods The effects of 10 Chinese herbal monomers against 20 strains of S. globosa were determined by micro-broth dilution method with reference to CLSI M38-A2 (3rd). Results The geometric mean(GM) of the minimum inhibitory concentration (MIC) of 10 Chinese herbal monomers ranged from 53.82 μg/mL to>1024 μg/mL. The effects of allicin, berberine, matrine, osthole and paeonol were the best, and the geometric mean were 53.82 μg/mL, 84.45 μg/mL, 103.97 μg/mL, 137.19 μg/mL and 284.05 μg/mL, respectively. Allicin was the most active antifungal against S. globosa, showing the range of MIC was 16-128 μg / mL. The minimum fungicidal concentration (MFC) ranged from 64 μg/mL to >1024 μg/mL. Allicin had the lowest MFC value and GM was 415.87 μg/mL. Conclusion Allicin, berberine, matrine, osthole and paeonol could against S. globosa, and the effects among Chinese herbal monomers were different.

Clinical features and treatment outcome of 30 cases of AIDS complicated with invasive cryptococcosis

TAO Peng-fei, YANG Xin-ping, LI Zheng-lun, ZHOU Qi-wen, MIN Hai-yan, QIAN Qing-jin

2020, 15(3):  150-153. 

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Objective To analyze the clinical characteristics of AIDS patients with invasive cryptococcus, and to provide reference for diagnosis and treatment. Methods The clinical data of inpatients diagnosed with AIDS complicated with invasive cryptococcosis in our hospital from January 2017 to December 2018 were retrospectively analyzed, and the data of clinical manifestations, laboratory examination, clinical treatment plan and outcome were systematically analyzed. Results A total of 30 patients were included. The clinical manifestations were fever, weight loss and fatigue. The CD4 T lymphocyte count was lower than 100cells/ L, and the white blood cells were significantly decreased by routine blood examination. The strain was sensitive to 5-fluorocytosine, amphotericin B, fluconazole injection and voriconazole. Lumbar puncture in patients with invasive cryptococcal meningitis indicated increased intracranial pressure, and cerebrospinal fluid biochemistry indicated low glucose, low chloride and high protein. Clinically, four treatment regimens were mainly used, among which amphotericin B combined with fluconazole was more common, and the average length of stay was more than 40d. Conclusion AIDS patients with invasive cryptococcosis infection of blood flow, patients with infection of blood flow with cryptococcus isolates have a good prognosis, patients with infection of blood flow with cryptococcus isolates have a high fatality rate, and high intracranial pressure is one of the prognostic factors. Clinical timely HAART is the key to good treatment and prognosis.

Clinical characteristics and pathogen of patients with candidemia

ZHANG Wen, LUO Ying, GUO Cheng-jie, XU Ying-chun, XIAO Meng, LEI Cheng-bin, ZHAO Yan, BI Lei

2020, 15(3):  154-160. 

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Objective To explore clinical features, pathogen distribution, antifungal resistance and prognosis of patients with candidiamia. Methods The clinical data of inpatients with candidemia from January 2011 to December 2018 in Zibo Central Hospital were retrospectively studied, and the risk factors, clinical features, species distribution, drug sensitivity and clinical outcomes were described and analyzed. Results The inpatients with candidemia were mainly from ICU (35.9%), and most of them were elderly patients (>60 years old, 75.7%). Candida albicans (40.8%) were the primary species, followed by C. tropicalis (20.4%), C. parapsilosis (20.4%), C. glabrata (10.7%) and C. krusei (3.9%). The separation rate of C. albicans showed the tendency of firstly increased and then decreased from 2011 to 2018, and non-albicans candidemia had risen in recent years. Azole resistance mainly arised in C. tropicalis and C. glabrata, and the sensitivity of C. albicans and C. parapsilosis to azole also declined. The 30-day crude mortality rate was 39.8%. Multivariate regression analysis revealed that agedness (OR=1.091, =0.001) and diabetes (OR=9.709, P=0.005) were independent risk factors, and targeted antifungal therapy (OR=0.142, P=0.008) could improve the prognosis of patients. Conclusion The incidence of candidemia can be reduced by treating basic diseases and reducing invasive operations in high-risk hospitalized patients, and the prognosis of patients can be improved by timely and targeted application of antifungal drugs.