Objective To determine whether Trichophyton rubrum isolated from different lesions in the same patient is of different strains. Methods DNA was extracted from the isolates, then subjected to a polymerase chain reaction-based typing which analyzed the number of repetitive elements in the non-transcribed spacer region of the ribosomal DNA gene repeats. Results Thirty-six strains of T. rubrum were isolated from 17 patients with fungal infection on multiple sites. All strains could be classified into 10 genotypes. The genotype distribution was unrelated to sites of infection. It happened to 8 of the 17 patients that multiple genotypes were involved in T. rubrum infection on different sites in the same body. Conclusion The study shows that multiple genotypes are involved in T. rubrum infection on different sites in the same patient, suggesting external sources of infection rather than infection from a different site in the same individual.

Objectives To analyse the antifungal susceptibility of Candida spp. isolated from the patients with recurrent vulvovaginal candidiasis (RVVC), vulvovaginal candidiasis (VVC) and asymptomatic carriers and to study the correlation between different Candida strains and antifungal susceptibility. Method According to the NCCLS-M27-A scheme, the antifungal susceptibility of Candida spp. isolated from the above different groups was measured. Results Almost all the MICs of C. glabrata and C. krusei to 8 antifungal agents were higher than those of C. albicans. The average MIC of C. albicans isolated from RVVC patients was higher than that from asymptomatic carriers. The resistant strains were mainly isolated from the RVVC group. No resistant strains against itraconazole, fluconazole, ketoconazole, econazole and nystatin was found in asymptomatic carriers. Conclusions These results indicate that more attention has to be paid to the low susceptibility of non-Candida albicans in the treatment of vulvovaginal candidiasis, and the resistant strains may result from long-term or irregular antifungal treatment.

Objective To develop a murine model for infection by Cladosporium carrioni. Methods A total of 72 ICR mice were equally divided into 4 groups, group A (healthy mice inoculated by C. Carrioni suspension of 1 × 108 cfu conidia mL-1, group B (immune-suppressed mice inoculated by C. Carrioni sus pension of 1 × 108 cfu conidia mL-1), group C (immune-suppressed mice inoculated by C. Carrioni suspen-sion of 1 × 106 cfu conidia mL-1), group D (healthy mice inoculated by sodium chloride solution). C. Car-rioni suspension or sodium chloride solution was subcutaneously inoculated into foot pads of mice. On day 7, 30 and 60 after inoculation, 6 mice were killed in each group followed by the measurement of thickness of foot pads, pathology and mycology of skin samples taken from foot pads. Results In group A, B and C, there were swelling, blackening, ulceration and crusts at the inoculation site of all mice, with a morbidity of 100%. The thickness of foot pads in group A on day 30 was significantly higher than that on day 7 (2.40 ± 0.45 mm vs 2.85 ± 0.47 mm, P < 0.05), but lower than that on day 60 (1.64 ± 0.13 ram, P < 0.05). In group B, increased thickness of foot pads was observed on day 30 compared with that on day 7 and day 60 (2.19 ± 0.27 mm vs 1.80 ± 0.21 mm and 1.86 + 0.22 mm, respectively, both P < 0.05), which was the case with group C (1.98 ± 0.06 nun vs 1.51 ± 0.11 mm and 1.82 ± 0.09 mm, respectively, both P < 0.05). No significant changes occurred to the thickness of foot pads in group D from day 7 to day 60 (P > 0.05). Pathological changes in group A, B and C included necrosis, abscess and chronic granuloma formation; dark brown sclerotic bodies were observed on HE and PAS staining as well as on direct microscopy; cultures of tissue samples grew Cladosporium carrionii. The mice in group D remained uninfected. Conclusion Mouse model for chromoblastomycosis may be established by subcutaneous inoculation of Cladosporium carrionii suspension into foot pads of healthy or immuno-suppressed mice.

Objective To investigate the characteristic,diagnosis and treatment of the nipple discharge.Methods The clinical data of 174 cases who were diagnosed as nipple dlscharge from January 2001 to December2006 were collected and analyzed retrospectively.Results Among those 174 cases,136 cases were confirmed histopathologieally to be intraduetal papilloma.The confirmed diagnosis rate of intraduetal papilloma by galaetograghy was 85.00%.Carcinoma-charge rate of the intraduetal papilloma was 5.17%.Conclusion Nipple discharge was the most common symptom in the intraductal papillomatesis.The galaetography was a valuable method in the diagnosis of the intraduetal papilloma.There was carcinoma-charge possibility in the intraduetal papilloma.

Objective To investigate the growth inhibition of Aspergillns fumigatus by Candida albicans in vitro and to develop the selective medium for clinical isolation of Aspergillus fumigatus.Methods Aspergillus fumigatus and Candida albicans were single or co-cultured in sabouraud dextrose agar(SDA) medium and SDA broth in dark at 25 ℃,and fungal growth,pigmentation,as well as colony diameter weredocumented.Results ①The sensitivity of culture of Aspergillus fumigatus and Candida albicans on SDAplate was 100CFU/ml.②The growth of 106CFU/ml and 103CFU/ml Aspergills fumigatus was completely inhibited by 106CFU/ml Candida albicans.③Growth inhibition of Aspergillus fumigatus was correlated with the concentration of Candida albicans.④SDA containing 1 mg/L fluconazole inhibited growth of Candida albicans,and no Candida albicans was detected on SDA containing 5 mg/L and 25 mg/L fluconazole.Growth of Aspergillus fumigatus was partially inhibited on SDA containing 25 mg/L fluconazole.Conclusions Candida albicans can inhibit the growth of Aspergillus fumigatus in vitro.SDA containing 5 mg/L fluconazole can be used as the selective medium for the isolation of Aspergillus fumigatus.

Objective To develop a murine model of chromomycosis by using Cladosporium carrioni and explore the pathogenicity of Cladosporium carrio nii in mice. Methods The suspension of Cladosporium carrioni was inoculated to two groups of mice, the immunocompetent mice and the immunosuppressed mice, b y intraperitoneal route using 1 ml inoculum containing 108 conidia/mL. All mice were sacrificed 30 days after inoculation, and then macroscopic examination, his topathology and fungal culture were performed. Results The morbidity in both g roups was 100% according to the dark brown hyphae and sclerotic bodies found in histopathologic examination and fungal culture. Macroscopic examination found th at the adhesion among the internal organs in immunocompetent mice was more sever e than that in immunosuppressed mice. Histopathologic sections showed that necro sis and inflammatory infiltration in immunocompetent mice were more obvious than those in immunosuppressed mice. Conclusions The virulence of Cladosporium car rionii strains is strong enough to construct experimental murine model of chromo mycosis, and animal passage of the strains is unnecessary. This murine model cou ld be used to study the pathogenesis of chromomycosis.

Objective Trichophyton rubrum strains can cause superficial infection and also deep tissue infection, but relevant animal model has not been reported yet.The aim of this study was to construct an animal model of granuloma infected by T.rubrum. Methods Three T.rubrum strains isolated from clinical granuloma tissues, 2 T.rubrum strains isolated from tinea infection and a standard strain of ATCCMYA4438 were selected.Corticosteroids were given to the Balb/C mice before and after the injection of the T. rubrum and mucin suspension and the mice model of granuloma was established.Direct microcopy, culture and histopathologic method were adopt to verify the infection effects. Results The mice inoculated with the T.rubrum granuloma strains with mucin suspension were examined after 21 days in the condition of applying appropriate dose of glucocorticoids.Direct microscopic examination showed the slender mycelium, fungal culture showed the growth of colony and histopathology showed excessive keratinization of foot tissue, formation of granuloma in the dermis with inflammatory cell infiltration of neutro-philic granulocyte and lymphocytes.However, the mice inoculated with the T.Rubrum tinea strains with mucin suspension showed the negative result. Conclusion The rubrum granuloma mice model can be es-tablished using the clinical isolates of T.rubrum granuloma strains with the mucin and glucocorticoids interventions.

Objective To analyze the gene expression of pathogenic factors in vaginal secretions of patients with vulvovaginal candidiasis by using Oligo chips. Methods RNA was extracted from vaginal secretions of 10 patients with vulvovaginal candidiasis and 3 asymptomatic carriers, and hybridized with oligonuscreened followed by a bioinformatic analysis. Results Comparing with the asymptomatic carriers, the patients showed a higher expression of 44 genes and lower expression of 17 genes. Of these differentially expressed (TLR) 4, HWP1, SAP2, SAP5, LIP4, EFG1 and CPH1 were highly expressed in more than 80% of the secretion samples from patients with an average ratio of 4.013, while LIP6 and WH11 were lowly expressed in more IFN-γ and TLR4 were associated with native immunity, HWP1 associated with hyphal adhesion and formation, SAP2, SAP5, LIP4 and LIP6 associated with extracellular hydrolysis, and EFG1, CPH1 and WH11 associated with phenotypic switching. Conclusions Both the host adaptive immunity deficiency and increased virulence of Candida species are involved in the pathogenesis of vulvovaginal candidiasis, and TLR4 possibly plays a certain role in the local immunity of patients with this entity.

Objective To determine the extracellular enzymatic activity of common pathogens for onychomycosis, in the hope of finding virulence factors associated with the pathogenesis of onychomycosis. Methods Strains tested in this study included standard strains of common dermatophyte and non-dermatophyte fungi as well as clinical isolates of Trichophyton rubrum from patients with onychomycosis. All the tested strains were cultured in medium containing nail fragments at 25 ℃ for 10 to 21 days followed by the determination of the nail fragment-containing medium, a significant increase was observed in the activities of esterase, esterase lipase, N-acetyl-β-glucosaminidase and α-mannosidase in dermatophytes compared with non-dermatophytes (all P ＜ 0.05 ), as well as in the activity of N-acetyl-β-glucosaminidase in Trichophyton rubrum compared with the other tested species of fungi (all P ＜ 0.05). No significant difference was noted in the activity of extracellular enzymes, except for that of naphthol-AS-BI-phosphohydrolase, between the isolates of Trichophyton rubrum from patients with different ranges of scoring clinical index for onychomycosis (SCIO). Conclusions In specific conditions, the extracellular enzymatic activity of fungi isolated from patients with onychomycosis is associated with fungal species, and may have a certain influence on the manifestations of anychomycosis.

Objective To estimate the application value of colony PCR in the detection of pathogenic filamentous fungi. Methods Colony PCR was established and performed to amplify the internal transcribed spacer (ITS) region of 19 species (strains) of filamentous fungus followed by sequencing analysis. At the same time, DNA extracts from 8 of the 19 species of filamentous fungus were subjected to conventional PCR. Hha I and Hinf I endonucleases were used for restriction fragment length polymorphism (RFLP) analysis of the conventional and colony PCR products. Comparison analysis was carried out between the colony and conventional PCR. Results Of the 19 strains, 16(84.2%) yielded positive results by colony PCR; sequence analysis of the PCR products of ITS region revealed a 96% - 100% similarity with the reference sequence (NCBI database)of corresponding fungi. The amplification product length and RFLP profile of these products from the 8 species of filamentous fungus, except for those from Aspergillus nidulans, were consistent between the colony and conventional PCR. Conclusions Compared with conventional PCR, colony PCR-based detection of filamentous fungi is easy to operate, time and labor-saving, with high accuracy and reliability, and can be applied to the rapid identification of filamentous fungi.