Table of Content

28 June 2018, Volume 13 Issue 3

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论文

Loss of phospholipase D in mice against Aspergillus fumigatus infection

LIU Xiao-yu, ZHANG Chang-jian, HU Ying-song, CHEN Fang-yan, HAN Li

2018, 13(3):  129-133. 

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Objective To reveal the important role and the possible mechanism of the host phospholipase D in response to the infection of Aspergillus fumigatus. Methods Wild type mice and phospholipase D double genes knockout mice (pld1^-/-pld2^-/-) were infected with Aspergillus fumigatus conidia by nasal instillation. After infection, the lung tissure burden of Aspergillus fumigatus was detected by tissue homogenization and CFU counting. The secretion of inflammatory factors in BALF were detected by multi factor detection kit from eBioscience Company. Bone marrow cells of two kinds of mice were isolated and differentiated into mature macrophages (BMDM), their ability to phagocyte and kill Aspergillus fumigatus was detected by the nystatin protection method. Results Six hours post infection, the load of pulmonary fungi in pld1^-/-pld2^-/- mice was significantly higher than that of wild type group; and a large number of Aspergillus fumigatus spores were found in BALF and alveolar macrophages of pld1^-/-pld2^-/- mice. Meanwhile, the concentration of inflammatory factors in BALF of pld1^-/-pld2^-/- mice was significantly higher. In vitro experiments showed that the phagocytosis and killing ability of BMDM of pld1^-/-pld2^-/- mice was significantly reduced. Conclusion The simultaneous knockout of the pld1 and pld2 genes in mice significantly decreased the phagocytosis and fungicidal ablities of the macrophage, which might interfere mice againsting A.fumigatus infection. Meanwhile, the pld1^-/-pld2^-/- mice might secrete a large number of inflammatory factors to recruit other immune cells to kill Aspergillus fumigatus conidia.

Evaluation of fungicidal effect of low frequency ultrasound and amphotericin B-loaded nanoparticles on C. albicans biofilm

YANG Min, XIE Shuang, DONG Yu, DU Yong-hong, LI Dai-rong

2018, 13(3):  134-138. 

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Objective To evaluate the synergistic fungicidal effect of low frequency ultrasound combined with amphotericin B-loaded nanoparticles (AmB-NPs) on C. albicans biofilm in vitro. Method AmB-NPs were prepared by a polylactic-co-glycolic acid (PLGA) double emulsion method. XTT reduction method was performed to evaluate the effect of ultrasound with AmB-NPs on the activity of mature biofilm. The morphology of biofilm was observed by laser scanning confocal microscope. The protease and phospholipase activities of biofilm were detected by UV spectrophotometer. Results The particle size and shape of AmB-NPs were uniform. The biofilm activity of C. albicans decreased significantly after the joint action of ultrasonic irradiation and AmB-NPs compared with the control group (P<0.01). Simultaneously, the biofilm became thin and loose, with significant decreased activities of protease and phospholipase aenzyme. Conclusion Low-frequency ultrasound combined with AmB-NPs showed significant synergistic antifungal effect against C. albicans biofilm in vitro.

In vitro study of the 5-aminolevulinic acid photodynamic therapy for Trichophyton tonsurans strain isolated from granuloma

WU Qian-qian, SHI Lei, ZHANG Hai-yan, ZHANG Ling-lin, GAO Zhi-qin, YANG Hong, YANG Lian-juan, WANG Xiu-li

2018, 13(3):  139-143. 

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Objective To demonstrate the fungicidal effect and action mechanism of 5-aminolevulinic acid photodynamic therapy (ALA-PDT) against Trichophyton tonsurans strain isolated from granuloma. Methods The clinical isolate of Trichophyton tonsurans granuloma was cultured in potato dextrose agar medium to prepare the suspension. After 6 hours of incubation, the production of PpⅨ was observed by fluorescence microscopy. Following PDT (at light energy density of 50, 100, 150, 175, or 200 J/cm²), the growth inhibitory of this T.tonsurans strain was observed by colony cultivation. Results Fluorescence microscopy showed obvious PpⅨ production of brick red fluorescent substance. At the light energy density of 175 and 200 J/cm², the significantly restricted growth of colonies was evidenced by decreased concentration of suspension. The suspension concentrations were (0.53±0.058)×10⁵ CFU/mL and (0.13±0.057)×10⁵ CFU/mL at light energy density of 175 and 200 J/cm² respectively, which were significantly lower than those of control group, single drug group, single light group and other low energy density ALA-PDT group (P<0.05). Conclusion ALA-PDT exerted fungicidal effect against Trichophyton tonsurans granuloma isolate, which might provide a theoretical basis for the clinical treatment of dermatophytes.

The effect of rifapentine and fluconazole on the cell cycle of FLC-resistance Candida albicans

WANG Yu-lian, YUAN-Gong, CHEN Ya-nan, CAO Yong-bing, WU Jian-hua

2018, 13(3):  144-147. 

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Objective To explore the effect of rifapentine and flueonazole on the cell cycle of FLC-resistance Candida albicans in vitro. Methods Candida albicans suspension were cocultured with rifapentine alone or combined with flueonazole for 12 h. A blank control group without drugs was established. The DNA content of Candida albicans in each group was detected by Flow Cytometrya after Propidium iodide (PI) staining. The effects of drugs on cell cycle of FLC-resistance Candida albicans including the rifapentine alone or combined with flueonazole were analyzed. Results Compared with growth control group and single drug group, DNA content decreased in G0/G1 phase (P<0.01) of combination group, but significantly increased in G2/M phase (P<0.01).This showed that cell cycle was inhibited at G2/M phase when rifapentine and fluconazole were conbined. Conclusion The results showed that the combination of rifapentine and fluconazole had a significantly effect on arresting of FLC-resistance Candida albicans'cell cycel and inhibiting its normal division and proliferation.

Clinical characteristics on 11 immunocompetent patients with pulmonary cryptococcosis and literature review

WANG Yan-ling, YANG Yu-heng, YU Nong, DENG Shu-wen

2018, 13(3):  148-151,163. 

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Objective To investigate the clinical characteristics of PC in 11 immunocompetent patients. Methods Eleven PC cases diagnosed histopathologically and mycologically from January 2004 to December 2016 were investigated retrospectively. General clinical data, radiology, way of diagnosis were analyzed. Results Eleven patients were immunocompetent. Seven PC patients (63.6%) had obesity, hypertension or coronary heart disease; one patient had smoking history for 20 years; only one case was found during checkup. Six cases were misdiagosised (54.54%,6/11), carcinoma of lung was the most misdiagnosised disease. The most common CT finding in those patients with pulmonary cryptococcosis was pulmonary solitary nodules with halo sign in middle and lowerlung predominance. Areas of hazy increased attenuation, and areas of cavitation were uncommon. Nine of 11 PC patients were diagnosised by histopathological examination; 2 of them found the Cryptococcus spp. in ink KOH test; one had positive serum cryptococcal antigen (CrAg). Conclusion PC could occurred in immunocompetent individuals with obesity, hypertension or coronary heart disease. Smoking could also be a risk factor. CT commonly demonstrated pulmonary solitary nodules in middle and upper lung. Definitive diagnosis could be made histopathologically. Better diagnostic outcomes could be achieved by bronchoscopy, bronchoalveolar lavage/trans-bronchial biopsy, or via thoracotomy-directed biopsy.

Molecular epidemiology and clinical characteristics of 34 cases of cryptococcal infection in HIV patients

CHEN Yan-hui, YU Feng, BIAN Ze-yuan, HONG Jian-ming, ZHONG Qiao-shi, HANG Ya-ping, ZHANG Nan, HU Long-hua

2018, 13(3):  152-157. 

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Objective To analyze the molecular epidemiology and clinical characteristics of cryptococcosis occurred in HIV-infected patients, and to provide evidence for the prevention and treatment of cryptococcosis. Methods A total of 34 Cryptococcus neoformans strains isolated from HIV-infected patients were collected and the general data of patients were investigated retrospectively. The isolates were first identified based on the matrix-assisted laser desorption/ionization time-of flight mass spectrometry (MALDI-TOF MS), the MICs of the five kinds of antifungal drugs were carried out according to ATB Fungus3. PCR with specific primers were used to identify variants and mating types. The clonal relatedness was characterized by multilocus sequence typing (MLST) analysis. Results Most Cryptococcus neoformans strains were isolated from middle-aged men and mainly came from cerebrospinal fluid (73.5%). Cerebrospinal fluid pressure was (27.26±11.52) mmH₂O on average,median CD4 T-cell count was 28 cells/μL (3~163 cells/μL), median CSF WBC count was 32 (2~110)×10⁶/L, the median protein was 362 mg/L (160~2 730 mg/L), median glucose 2.28 mmol/L (1.50~5.98 mmol/L). All strains were sensitive to five kinds of antifungal drugs and were identified as serotype A, mating type α and molecular type VNⅠ. MLST analysis divided the 34 isolates into two sequence types (STs), including ST5 (n=32), ST32 (n=1) and ST186 (n=1). Conclusion In recent two years, cryptococcosis occurred in HIV-infected patients were more common in middle-aged men. The laboratory tests lacked specificity. The resistance of Cryptococcus neoformans isolates from HIV-infected patients was not serious, ST5 is the main clonal.

Central nervous system phaeohyphomycosis: literature review of 203 cases

GE Gai, LIANG Guan-zhao, SHI Dong-mei

2018, 13(3):  158-163. 

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Objective To review 203 central nervous system phaeohyphomycosis (CPHM) cases at home and abroad to help early diagnosis and reduce mortality. Methods Sex composition, age trait, geographical features, early clinical manifestations, associated diseases, therapies and outcome in 203 CPHM cases before August 2017 in home and abroad were collected. Results CPHM occured more commonly in immunocompetent patients than in immunedebilitated patients, predoiminantly in young and adult males. Cladophilophora bantiana was the most common etiologic agent followed by Ramichloridium mackenziei and Exophiala dermatitidis. There were no early specific symptoms of CPHM,except increased intracranial pressure. The combination of antifungal therapy and surgical resection could improve survival. However, few knowledge of CPHM and lacking specific detecting method might led to misdiagnosis or delayed treatment. An overall mortality was 80%. Conclusions With untypical features, CPHM had high mortality due to delayed diagnosis and the lack of standardized therapy.

Efficacy of 1% sulconazole nitrate spray in the treatment of tinea pedis and preventive effect of spraying the shoes and socks

WANG Mei-fang, LI Lin-feng, CHENG hai-yan, CHEN Ze-yu, TIAN Ke

2018, 13(3):  164-167. 

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Objective To evaluate the efficacy and safety of sulconazole nitrate spray 1% in the treatment of tinea pedis and to observe the effect of spraying the socks and shoes on preventing the disease from recurrence. Methods One hundren and twenty patients diagnosed as tinea pedis were randomly divided into two groups, being treated with sulconazole nitrate spray 1% and miconazole nitrate cream 2% respectively.The clinical efficacy was evaluated after two weeks and four weeks. 30 clinically cured patients would continue to spray their shoes and socks to control the local environmental fungi for one month. Then the recurrence rate was compared with that of the other 30 patients without any further treatment. Results Clinical efficiency of experimental and control groups were 90.48% and 49.23% respectively after 4 weeks. There was no significant difference between the two groups (P=0.085). The recurrence rates of the group who had continued to spray the socks and shoes and the group who had not continued to use drugs were respectively 10% and 26.7%.The difference was statistically significant (P=0.038). Conclusion This study suggested that sulconazole nitrate spray 1% was effective and safe in the treatment of tinea pedis. Spraying the socks and shoes with sulconazole nitrate spray 1% would reduce the recurrence of tinea pedis.

Clinical significance of plasma (1-3)-β-D glucan for diagnosis of invasive fungal infection

Shang Yuan-yuan, Yu Nan, Dong Ling-di, Li Sha-sha, Liu Yu-xi

2018, 13(3):  168-172. 

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Objective To explore the diagnostic value of plasma (1-3)-β-D-glucan test in the nosocomial invasive fungal infection (IFI). Methed The MB-80 microbial dynamics detection system was used to conduct the G test in 132 inpatients with suspected nosocomial IFI in the General Hospital of NingXia Medical University from September 2016 to April 2017. Meanwhile, the result of the conventional fungal culture and the result of combined detection were analyzed by ROC curve. Results Among 132 cases, 43 cases were diagnosed as IFI. The sensitivity of fungal culture was 67%,the specificity was 96%, PPV and NPV were 0.72 and 0.80. The sensitivity of G Test was 70%,the specificity was 88%, PPV and NPV were 0.73 and 0.86. Combined the fungal culture and G test, the sensitivity was improved to 84%,the specificity was improved to 92%. The Youden exponents of fungal culture,G test and combined detection were 0.63,0.58 and 0.76. The areas under the ROC curves of fungal culture,G test and combined detection were 0.757,0.717 and 0.798. Conclusion G test was a simple, rapidly practical early diagnostic method of fungial infection. The combined detection could increase the sensitivity,specificity and diagnostic value for diagnosing nosocomial IFI.

Analysis on pathogenic distribution and sensitivity of nosocomical candidemia

HUA Hai-kang, LAI Mei-ling, HU Ren-jing, ZHANG Hai-ping, ZHOU Cheng-long, ZHANG Xiao-li, GE Xiao-li

2018, 13(3):  173-176. 

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Objective To investigate the pathogen distribution and drug resistance of Candida bloodstream infection. Methods The data of Candida isolated from blood culture were retrospectively analyzed in the Wuxi No.2 People's Hospital from 2013 to 2016. Results Fifty-six pathogenic Candida strains were isolated, which included Candida albicans (57%,32/56), Candida parapsilosis (20%,11/56) and Candida glabrata (15.8%,9/56). The patients mainly came from the ICU (23.2%,13/56) and the department of Nephrology (17.9%, 10/56). The difference of sensitivity rates between Candida albicans and non-albicans candida was notable, which were 93.8% and 62.5%. The sensitivity rates of 56 Candida strains to amphotericin B and flucytosine were 100%. The sensitivity rates of Candida glabrata and Candida parapsilosis to fluconazole were 55.6% and 72.7%. All the Candida krusi strains were resistance to fluconazole. Conclusions The main pathogenic Candida isolated from bloodstream infection was Candida albicans. The sensitivity tests showed that non-albicans candida species had relatively strong endurance to fluconazole. Therefore, drug resistance should be considered in the selection of antifungal drugs.

Pathogens distribution and risk factors of pulmonary infection in elderly patients with acute exacerbation of chronic obstructive pulmonary disease

WEI Yong-gang, YAN Hong-ying, XIAO Yuan-hong

2018, 13(3):  177-180,183. 

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Objective To investigate the distribution of pathogens and risk factors of acute exacerbation of chronic obstructive pulmonary disease (COPD) in elderly patients with pulmonary infection. Methods Two hundred and sixty elderly patients with acute exacerbation of chronic obstructive pulmonary disease in our hospital were selected. All patients were divided into infection group and non-infection group accorded to the occurrence of pulmonary infection. The clinical data of all cases were recorded, and the pulmonary infection patients were given analysis of the risk factors and the pathogens distribution. Results There were 30 patients of pulmonary infection in the 260 patients (11.5%). In the infection group, a total of 45 strains of pathogen including 25 strains of gram negative bacteria, 18 strains of gram positive bacteria, 2 strains of candida spp. were isolated. The top 4 pathogens were Pseudomonas aeruginosa, Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus aureus etc. The Pseudomonas aeruginosa, Klebsiella pneumoniae had relatively high rate of resistance to erythromycin, gentamicin, and the Pseudomonas aeruginosa, Staphylococcus aureus had relatively high rate of levofloxacin and meropenem. Univariate regression analysis showed that the history of diabetes, smoking, long-term use of antibiotics, long-term use of hormones, long-term bed were significant different in the infection group and in non-infection group (P<0.05); Non conditional Logistic regression analysis showed that the diabetes (OR=10.593, P=0.000), smoking (OR=6.194, P=0.008), long-term use of antibiotics (OR=3.255, P=0.014) were the risk factors of pulmonary infection (P<0.05). Conclusion Pulmonary infection was common in acute exacerbation of chronic obstructive pulmonary disease, Gram-negative bacteria might be the main pathogen. Diabetes, smoking, long-term use of antibiotics were the risk factors of pulmonary infection.

Monilial granuloma caused by Candida parapsilosisis: a case report

WEN Yi-yang, CHEN Yu-chong, CAI Qing, YANG Lian-juan

2018, 13(3):  181-183. 

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A case of monilial granuloma caused by Candida parapsilosisis was reported. A-15-year-old boy had a history of rash on the face and neck for 2 months. Physical examination showed 3 annular plaques with diameters of 1-3 cm on his right face and neck. The margin of the lesions was edematous and swollen, and the centre was necrotic and crusted. There was a slight yellow exudation on the surface. A biopsy of neck lesion revealed pseudoepitheliomatous hyperplasia of epidermis, diffuse inflammatory cell infiltration into the dermis, mainly in lymphocytes, eosinophils, and plasma cells, with local necrosis and vascular proliferation. The isolate from the biopsy specimen was identified as Candida parapsilosisis. The patient was discharged on oral itraconazole 100 mg twice daily and external bifonazole cream for 6 week, while using dermoscopy to evaluate the efficacy of treatment, the lesions were slightly improved. The patient is still under treatment.

The current therapeutics and research progress of topical antifungal agents

SONG Na-na, ZHOU Xiao-wei, LI Xiao-fang, LIU Wei-da

2018, 13(3):  184-187. 

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Onychomycosis is widespread with a low initial treatment rate, high recurrence rate and high re-infection rate. The commonly used methods include topical and oral medications. Due to the side effects and drug-drug interactions of oral antifungal agents, it is mainly focused on the development of topical antifungal antimycotics in recent years. The mechanism, efficiency related to experiments in vitro or vivo and research progress of topical antifungal formulations including traditional drugs (amorolfine and ciclopirox), novel drugs (efinaconazole and tavaborole) and drugs under development (terbinafine hydrochloride, luliconazole, ME1111 and tazarotene) are reviewed in this paper.