Table of Content

28 February 2019, Volume 14 Issue 1

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论文

Clinical and laboratory characteristics of Candida bloodstream infection

CAO Yun, SONG Shao-ting, LI Shuai, FANG Wen-jie, LIAO Wan-qing, PAN Wei-hua, XIONG Lin

2019, 14(1):  7-10. 

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Objective To analyze the pathogenicity and drug susceptibility test based on blood culture fungal positive specimens, and to explore the clinical and laboratory characteristics of Candida in bloodstream infection. Methods The clinical data of blood culture were collected from June 2012 to September 2018 in our hospital. The fungal species, positive alarm time (TTP), specimen delivery department, pathogen source and fluconazole susceptibility test results were analyzed. Results Eighty-six strains of fungi were isolated from blood culture, 50 strains were non-repetitive isolates, and 44 strains were Candida spp. The top three Candida species were Candida albicans with 18 strains(40.91%), Candida parapsilosis and Candida glabrata with 10 strains respectively (22.73%); and Candida parapsilosis TTP[(56.14±7.22)h] is the longest, Candida tropicalis TTP[(19.36 ± 1.24) h] is the shortest, Candida albicans[(34.67 ± 2.98) h] and Candida glabrata[(38.16 ± 4.12) h] TTP in the middle; The top three departments of candidiasis was neurosurgery with 10 strains (22.73%), tumor hematology with 6 strains (13.64%), and intensive care unit with 5 strains (11.36%);urinary system was the most common source of infection, followed by abdominal drainage infection and catheter infection; Candida and Candida tropicalis were sensitive to fluconazole at 100%, Candida albicans and Candida glabrata were 88.89% and 70%, respectively, one strain of C.kruseii was resistant and one strain of Saccharomyces cerevisiae was sensitive. Conclusion The most common bloodstream infection of Candida in hospital is Candida albicans, followed by Candida parapsilosis and Candida.

Preparation of recombinant proteinphosphopantetheinyl transferase Ppt2 of Candida albicans

MENG Ling-ning, LIU Jin-yan, ZHAO Yue, WANG Yu-ting, XIANG Ming-jie

2019, 14(1):  11-15. 

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Objective To prepare the phosphopantetheinyl transferase Ppt2 of Candida albicans, which would be used for the drug screening of Ppt2 as the target by fluorescence polarization. Methods The DNA of PPT2 was amplified by PCR using Candida albicans BWP17 genomic DNA as a template and linked with the linearized vector pET43.1a(+) by homologous recombination. The recombinant plasmid pET43.1a(+)/PPT2 was successfully constructed and transformed into E.coli BL21 (DE3) competent cells with IPTG. The protein Ppt2 was further extracted using ultrasonic fragmentation and verified solubility by SDS-PAGE. The column Ni was used to purify the protein and the elution fraction was dialyzed to enrich the target protein. The protein Ppt2 was finally determined by the Bradford method. Results The recombinant protein Ppt2 was highly expressed in E.coli BL21 with final concentration of 0.4 mg/mL. The results of SDS-PAGE electrophoresis showed that Ppt2 had both soluble form and inclusion body form. After amplification of the amount of E.Coli BL21, the soluble protein was purified by column Ni to obtain the target protein. After dialysis and enrichment of protein Ppt2, it was measured by Bradford method that protein Ppt2 concentration is 0.4 mg/mL. Conclusion The phosphopantetheinyl transferase protein Ppt2 of Candida albicans was successfully prepared, which could provide the basis for the future drug screening of Ppt2 as a target by fluorescence polarization.

The distribution and drug resistance rate of 3759 cases of fungal keratitis

WANG Qian, LUO Dan, CHEN Hua-bo, SUN Xiao-yan

2019, 14(1):  16-19. 

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Objective To investigate the pathogenic fungi distribution and drug resistance rate of fungal corneal infection, and to provide reference for clinical diagnosis and treatment. Method Retrospectively analyzed 3759 cases of keratitis patients in Qingdao eye hospital from January 2014 to December 2017. The keratitis patients datas were selected for fungal culture and drug resistance test, the results were collected and analyzed. Results The total positive rate was 22%(828 cases). Most of patients were old men, the most common fungi were Fusarium sp(41%), followed by Alternaria sp(30%). The positive rate of fungal culture in the fourth quarter of the year was significantly higher than those in the other quarters. The drug resistance rates of voriconazole, amphotericin B, itraconazole, ketoconazole and fluconazole were 1.2%, 1.6%, 17.6%, 32% and 96.1%. Conclusion Fungal infection of the cornea was on the rise and seasonal variation. The majority pathogens of fungal keratitis were Fusarium sp,followed by Alternaria sp. The drug resistance rates of fluconazole and ketoconazole were higher than that of other three antifungal drugs.

Clinical analysis of 12 cases with cutaneous mucormycosis caused by Rhizopus arrhizus

LIU Jia, LIANG Guan-zhao, YANG Jia, GE Li-yu, DONG Jia-cheng, LIU Wei-da

2019, 14(1):  20-23. 

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Objective To analyze clinical features of cutaneous mucormycosis caused by Rhizopus arrhizus since 1991 in China, and to further explore the typical clinical manifestation, predisposing factors, treatment strategies and prognosis. Methods Cases infected with rhizopus arrhizus reported in various data bases of China were searched thoroughly, clear diagnostic evidence should be described. Results Twelve cases were included in this paper. There were 10 males and 2 females separately, the average age of those patients was 46.83 years old. Uncontrolled diabetes mellitus was the most common risk factor and traumatic history could always be found in most cases. Aggressive erosion and ulcer were the most common clinical manifestations, and black eschar could be served as a clue to diagnosis. Combination amphotericin B with topical debridement was the most common treatment. Of all the patients, 3 of them resulted in death, 9 patients received favorable results of antifungal agents, but 2 cases underwent amputation surgery pitifully. Conclusions Rhizopus arrhizus was one of the most common pathogens causing mucormycosis, which might lead to death and deformity. Early diagnose and individual treatment are of great importance in saving lives and improving life qualities.

Pulmonary aspergilloma: a case report and laboratory study

SHAO Jin, YU Jin, WAN Zhe, LI Ruo-yu, WANG Ai-ping

2019, 14(1):  24-27. 

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Objective Laboratory study was performed on the surgical specimens of a pulmonary fungus ball patient without fungal culture to determine the pathogen. Methods A patient with hemoptysis was found lung space occupying lesion on left upper lung and received left upper lobectomy. The postoperative histopathologic diagnosis was pulmonary fungus ball (The pathogenic fungi was the most likely Aspergillus) and the tissue was not used for fungal culture. In this study, fungal fluorescence staining, immunohistochemistry and tissue PCR assays were carried on histopathological specimens. Results Fungal fluorescence staining showed bright blue septate mycelium, with sharp-angle branch. The results of immunohistochemistry suggested that the pathogen was Aspergillus. Tissue PCR result was negative. Conclusion When lacking of fungal culture results, immunohistochemistry techniques might help determine the genus of pathogenic fungi, which was helpful for clinical diagnosis and treatment.

The clinical features of chronic pulmonary aspergillosis in 55 patients

KANG Wei-fang

2019, 14(1):  28-31. 

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Objective To study the clinical features of chronic pulmonary aspergillosis(CPA),so as to provide evidence for early diagnosis and treatment. Methods A retrospective analysis was made upon clincial data, imaging features and laboratory tests of 55 patients with CPA. Results Of the 55 patients diagnosed with CPA, 34 patients were diagnosed with chronic cavitary pulmonary aspergillosis,15 patients with semi-invasive aspergillosis,4 patients with simple aspergilloma, and 2 patients with aspergillus nodule. There were 29 cases of chronic obstructive pulmonary disease(52.7%),6 cases of malignant tumor(10.9%),5 cases of pulmonary tuberculosis(9.1%),4 cases of asthma(7.3%),4 cases of bronchiectasis(7.3%) and 1 cases of interstitial lung disease(1.8%) in CPA. The most common symptoms in the CPA patients were cough(94.5%),sputum(81.8%),hemoptysis(54.5%),and fever(36.4%).The most common computerized tomography abnormalities were cavities(96.4%),nodule(63.6%),infiltration or consolidation(7.3%),and pleural thickening(5.5%). Conclusion CPA patients were usually associated with chronic obstructive pulmonary disease, malignant tumor, pulmonary tuberculosis, asthma, bronchiectasis and interstitial lung disease. Chronic cavitary pulmonary aspergillosis was the most common types of CPA. Actively searching for etiological and imaging evidences, including pneumocentesis or pulmonary resection if necessary, early antifungal treatment would improve prognosis of patients with CPA.

The efficacy of Rishuan lotion combined with clotrimazole on treatment of vulvovaginal candidiasis in mice

LIU Dao-fan, WU Qi-kang

2019, 14(1):  32-36. 

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Objective To explore the curative effect of Rishuan lotion (the main component includes kushen) combined with clotrimazole on the treatment of murine vulvovaginal candidiasis. Methods A total of 95 BALB/c female rats (8 weeks old) were randomly divided into four groups, including group A (n=5, blank control group), group B (n=30, model control group), group C (n=30, clotrimazole group) and group D (n=30, Rishuan lotion plus clotrimazole group). Mice in group B,C and D were inoculated with Candida albicans vagina. Mice in group A and B were given equal amount of normal saline to wash the vagina, while mice in group C and D were treated with clotrimazole, Rishuan lotion combined with clotrimazole, respectively. The weight changes of mice in each group were observed before and during the treatment. In group B,C and D, the vaginal lavage fluid was cultured at 1, 2, 3, 6, 8, 10, 14 and 16 days. The vaginal fungal load of mice was detected by coating method, and the levels of IL-2 and IL-8 were detected by ELISA. The expression of NLRP3, ASC and caspase-1 in the vaginal tissue of mice was detected by qRT-PCR. Results At the first 8 days, the weight of mice in the four groups remained stable. After inoculation, the weight of mice in group B,C and D decreased, and then gradually increased, which was close to the level before inoculation. Compared with group A, vaginal fungal load increased in group B, C and D. But there was a statistical difference in vaginal fungal load between group B, C and D. Vaginal fungal load in group D was less than that in group B and group C, while the load in in group C was less than that in group B. Compared with blank control group, IL-2 and IL-8 increased in vaginal lavage fluid of group B, C and D. However, there was a significant difference in the levels of IL-2 and IL-8 in group B, C and D after treatment. The levels of IL-2 and IL-8 in group D were significantly lower than those in group C, and the levels of IL-2 and IL-8 in group C were significantly lower than those in group B (P<0.01). Compared with the blank control group, the expression of NLRP3, ASC and caspase-1mRNA in vagina tissue of group B, C and D was up-regulated. But there was a statistical difference among the three groups. The mRNA expression level in group B was higher than that in group C and group D, and the expression level in group C was higher than that in group D (P<0.01). Conclusion The effect of Rishuan lotion combined with clotrimazole on the treatment of murine vulvovaginal candidiasis was significant.

Candida infection and drug resistance in a hospital from 2015 to 2017

TIAN Huan

2019, 14(1):  37-41. 

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Objective To explore the species distribution and drug resisitance of Candida infection in our hospital from Jan. 2015 to Dec. 2017, and to provide etiological basis for clinical rational use of antifungal agents. Method All the data of patients with Candida infection including gender, age, organism, specimen type, department distribution and drug residence from Jan. 2015 to Dec were retrospectively collected. Result Total 720 Candida isolates mainly came from Geriatric department and Pulmonary department. C. albicans was the most common one, followed by C. tropicalis and C. parapsilosis. The main site of infection was respiratory tracts. There was no difference in the distributions of species from 2015 to 2017. The resistance of Candida albicans to fluconazole, voriconazole, itraconazole and amphotericin B was lower than that of non-Candida albicans (P<0.001). There were 102 multi-drug resistant strains in 720 Candida isolates, among which C. glabrata and C.krusei had the highest multi-drug resistance rates. Conclusion Candida albicans was still the main isolate of Candida, and there is no significant change in the isolation rates during three years; the drug resistance of Candida albicans and non-Candida albicans was different, and the multi-drug resistance rate of non-Candida albicans was higher than that of C. albicans. Clinical antifungal therapy should be based on the results of antifungal susceptibility test.

Tinea incognito on the neck with long-term misdiagnosis

ShaoXia, QiaoJian-jun

2019, 14(1):  45-46. 

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A 35-year-old male presented with rash on the neck with itching for 2 years. He had been diagnosed as dermatitis, eczema, pityriasis rosea, etc.. Topical glucocorticoids were not effective, and the lesion was enlarging. Physical examination revealed scaly papules, plaques and pustules on the neck. Under microscopy, a large number of fungal hyphae were observed by taking scales samples from the lesion. On fungal culture, spindle-shaped macroconidia were detected, which was consistent with Microsporum canis. A diagnosis of tinea incognito was made. It was cured after antifungal therapy.

A case of sporotrichosis in the uterine cervix

ZHANG Yin-an, LIU Xiao-na, YANG Jing, LIU Yue-jiao, SHI Ming, XUE Wen-cheng

2019, 14(1):  47-49. 

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Objective To investigate the clinical characteristics of sporotrichosis in the uterine cervix caused by Sporothrix schenckii). Methods A case of Sporothrix schenckii in the uterine cervixhad been analyzed. Review of major related literatures at domestic and foreign in recent years was also performed. Results Microscope examination of vaginal secretions showed lumpy, Gram-positive, round-shaped and oval-spores, which were larger than normal Gram-positive coccus and smaller than yeast-like fungal spores. Blood plate culture at 37℃ showed white colony with marginal edges, dry and wrinkled surface. It was not easy to pick up sample from the tough surface. The Gram stain of this white colony showed Gram-positive yeast-like spores. The, filamentous fungus-like colonies with white folds in centerhas been formed at 25℃ in the SDA culture. These characteristics were consistent with clinical laboratory diagnostic features of Sporothrix schenckii infection. Conclusion The cervix infection caused by Sporothrix schenckii was rare, and the characteristics of culturing colonies under different conditions would be helpful to timely diagnosis in the laboratory.

Application of guided course in fungal dermatology

YANG Ya-li, CHEN Jun, SUN Qi-ling, WANG Xue, Liu Ke, ZHANG Zhen, SHEN Zheng-yu, XU Hui

2019, 14(1):  50-52. 

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Guided course is an important part of the teaching of dermotology. It's main function is to stimulate students' interest in learning and cognition of dermotology through various forms of instructional design, and to inspire and guide students' clinical diagnosis and treatment thinking. The introduction of the follow-up subject teaching content plays a good role in linking up and down.Our group has long been committed to this teaching work, especially in the teaching of fungal skin diseases, to obtain satisfactory teaching feedback.