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Objective To evaluate the role of complement receptor 3 (CR3) on murine macrophages in the recognition of Penicillium marneffei.Methods RAW264.7 murine macrophage cells were cultured in vitro,and divided into four groups to be cocultured with inactivated and live Penicillium mameffei yeast cells as well as inactivated and live Penicillium marneffei conidia respectively at 37 ℃ in 5％ CO2 for one hour.The RAW264.7 cells incubated with phosphate-buffered saline (PBS) served as the blank control group.Then,reverse transcription-PCR was conducted to detect CR3 mRNA expression,Western blot to measure CR3 protein expression,flow cytometry to determine phagocytosis rate,enzyme-linked immunosorbent assay (ELISA) to quantify cytokine levels in culture supernatant.Some RAW264.7 macrophages were transfected with a specific siRNA targeting CR3 gene and cocultured with inactivated Penicillium marneffei conidia,subsequently,phagocytosis rate and supematant cytokine levels were determined.Data were processed by the SPSS 16.0 software,and one-way analysis of variance (ANOVA) was conducted for inter-group comparisons of these parameters.Results No significant differences were observed in the mRNA or protein expressions of CR3 among the four groups of RAW264.7 cells cocuhured with different forms of Penicillium marneffei (both P ＞ 0.05).The phagocytosis rate was 95.14％,89.56％,91.03％ and 90.78％ in RAW264.7 cells cocultured with inactivated conidia and yeast cells,as well as live conidia and yeast cells of Penicillium marneffei,respectively (P ＞ 0.05).The levels of interleukin (IL)-2,interferon (IFN)-γ,IL-4 and IL-10 in culture supernatant were increased at different degrees after one-hour coculture in the four coculture groups compared with the blank control group,but no statistical difference was noted among the four coculture groups in the supernatant levels of these cytokines (all P ＞ 0.05).After coculture with inactivated Penicillium marneffei conidia,the siRNA-transfected RAW264.7 cells showed a statistical decrease in phagocytosis rate (10.89％ vs.92.78％,P ＜ 0.05) and supernatant levels of IL-2,IFN-γ IL-4 and IL-10 compared with untransfected RAW264.7 cells.Conclusions In early stage of innate immunity,CR3 on macrophages may be one of the pattern recognition receptors participating in the recognition and mediation of phagocytosis of Penicillium marneffei.It's possible that both Thl-and Th2-type cytokines,such as IL-2,IFN-γ,IL-4 and IL-10,are involved in the immune response of macrophages against Penicillium marneffei.
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Objective To make a clinical and mycological analysis of tinea capitis in Guangzhou region. Methods A retrospective analysis was performed on 241 cases of tinea capitis collected from Feb, 1997 to Aug, 2010 in the Department of Dermatology, Sun Yet-sen Memorial Hospital. Results Among the 241 cases, 179 (74.27%) were tinea alba, 34 (14.11%) tinea kerion, 28 (11.62%) black dot ringworm, and no favus was observed. The dominant pathogenic fungi in decreasing order were Microsporum canis (182,80.89%), Trichophyton violaceum (25, 11.11%), Trichophyton mentagrophytes (10, 4.44%), Trichophyton tonsurans (3, 1.33%), Trichophyton rubrum (2, 0.89%), Microsporum gypseum (2, 0.89%) and Trichophyton verrucosum (1, 0.44%). Children were the main population (39.00%) suffering from tinea capitis. Conclusions In Guangzhou region, tinea alba is the most common type of tinea capitis, Microsporum canis is the main causative pathogen, and children are the predominate population affected by tinea capitis.
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Objectives To identify the route and time of transmission by Candida species from mothers' vagina to their neonates' mouth.Methods Specimens for fungal cultures were obtained from vaginal discharge of mothers just before delivery and also from the mouth of their offspring just after birth.Eleven mother-infant pairs were investigated.Candida species was identified based on morphology,biochemical analysis,and sequencing of D1/D2 domain of the large subunit of ribosomal DNA (LSUrDNA).Electrophoretic karyotyping (EK) and random amplified polymorphic DNA analysis (RAPD) were performed to search for DNA homology.Results Candida isolates (16 strains) from 8 mother-infant pairs were identified as Candida albicans by 100% homology of their D1/D2 sequences with reference strain C.albicans Y-12983 (GenBank access No.U45776).Similarly,4 strains from two mother-infant pairs and 2 isolates from the other pair were identified as Candida glabrata and Candida krusei,respectively,by 100% homology in sequences alignment of the domains with reference strains,C.glabrata Y-65(U44808) and C.krusei Y-5396 (U76347).The same EK profiles were found for each C.albicans or C.krusei strain pair from both mother and her neonate.Although different EK bands with various molecular size were generated for each C.glabrata isolate pair,they were still considered to be homologous based on the fact that main EK bands were identical.Each isolate pair from mother and her infant presented almost the same RAPD profile,except for one pair,isolates F7n and F7m,which showed minor diverse DNA bands.Conclusion Eleven Candida isolates from neonates have identical molecular characteristics with their mother's isolates.Vertical transmission may be the main pathway of Candida spp.from mothers to their neonates.
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Objective To analyze the incidence of the disease, clinical features, diagnostic criteria, therapy and prognosis of Penicilliosis marneffei found in Guangdong province. Methods To analyze patients data, clinical features, laboratory findings, response to therapy, and prognosis of 15 cases Penicilliosis marneffei found in Guangdong province of China. Results The male was predominant compared with the female (ratio 2 to 1) and without occupational preference, but the patients with AIDS as underlying disease were mostly drivers and the unemployed. Thirteen patients were immunocompromised such as AIDS, connective tissue disease, and kidney transplant. Clinical features showed different manifestations, such as high fever, loss of weight, skin lesion, and respiratory system symptoms. Biopsy of the skin lesion showed PAS stain positive yeast-like, or sausage-form spores. Four patients were localized infection of the skin, eleven patients were systemic infection. Nine patients died, five recovered, 1 patient refused to be treated. Fifteen isolates from different anatomic sites of the patients were identified to be Penicillium marneffei by morphology and dimorphism in the culture, and eleven isolates among these 15 isolates were also confirmed by DNA sequence analysis. Conclusion The incidence rate of Penicilliosis marneffei become higher in the recent years and many patients were accompanied with AIDS in Guangdong province. Attention should be paid to the disease.
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Objective To investigate the relationship between the concentration of plasma (1→3)-?-D-glucan (?-D-glucan) and deep fungal infection. Methods Thirteen patients were recruited in this study, who were suspected with deep fungal infection. G-test TE reagent for ?-D-glucan measurement was used to detect the plasma (1→3) ?-D-glucan in the patients by using UV-2450 spectrophotometer at 545 nm wavelength. The final concentrations were calculated according to concentration conversion formula. Results Nine of thirteen patients were confirmed as deep fungal infection by positive tissue culture, in whom high concentrations of ?-D-glucan were detected, the highest concentration was 352.94 pg/mL (mixed infection), with a mean value of 203.47 pg/mL. In the other four patients with negative culture, the ?-D-glucan concentration was over 54.40 pg/mL in three patients and 16.16 pg/mL in the another. Our results showed that the sensiti-vity, specificity, positive predictive value and negative predictive value of this test were 92.31%, 100%, 100% and 98.36%, respectively. Conclusion G-test TE method is a simple and rapid test and may be used for the diagnosis of patients with deep fungal infection.