Objective To isolate and identify pathogenic f ungus in a patient with intracranial infection.Methods Specimens were taken from the spinal fluid of the patient.Then,microsco py and fungal culture were done to identify the pathogen.The hi stopathologic features were reprod uced through animal pathogenicity s tudy in a mice model.Results According to the colony appearance i n culture medium,the morphological features in microscopy,such as conidia arrangement and size,germ tube forming site,this fungus was identified as Bipolaris spicifera.Hyphae and swollen hyphal cells resembling chlamydospores,septate pi gmented hyphae were observed in brain tissue specimen of mice experimental model,which were consiste nt with phaeohyphomycosis.Conclu-sion This is the first case of phaeohyphom ycosis caused by Bipolaris spicifera reported in China.

Eleven salt-losers (SL group) and eleven non-salt-losers (NSL group) of congenital a-drenal hyperplasia due to 21-hydroxylase deficiency (21-OHD) were evaluated for sodium status under routine regimen of glucocorticoid (4 salt-losers with mineralocorticoid at the same time). Venous samples were collected for the measurement of plasma renin activity (PRA), aldosterone (Aldo) , and a-drenocorticotropin (ACTH), and for serum cortisol (F), 17-OH progesterone (17-OHP), and testosterone (T). Urine samples were collected for 17-ketosteroid (17-KS). 10 age-matched normal children were chosen as control. The results showed that concentration of PRA, Aldo, 17-OHP, and T, were higher in both groups than in normal subjects (P

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.

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 evaluate and monitor the efifcacy of GnRH analogs (GnRHa) therapy. Methods Thirty girls with central precocious puberty diagnosied by LHRH stimulation test were treated with GnRHa for 6-24 months. The LHRH stimula-tion test were performed again at 3 months after initiation of therapy and then every 6 months during treatment. The relationship of peark LH and clinical suppressing pubertal (including Turner stage, bone age, grwoth speed) were compared. The monitor effect of peak LH to efifcacy of GnRHa were eveluated. Results Ninety LHRH stimulation tests were performed, only 7 cases were found to have clinical pubertal development. After 6 months treatment, the base LH level of thirty girls (0.48±0.20) IU/L was signiifcantly lower than that before the treatment (0.75±0.35 IU/L) (P=0.000). The correlation coefifcient between base LH and peak LH was 0.62. The best correlation between clinical suppressing pubertal and LHRH stimulation test was achieved when peak LH was less than 2 IU/L (85.7%sensitivity, 100%speciifcity). Conclusions Base LH value can be used in preliminary as-sessment of the efifcacy of GnRHa therapy for girls with central precocious puberty. The peak LH less than 2 IU/L can be as the indicator of treatment efifcacy.

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 compare broth microdilution and agar dilution methods for in vitro testing of activities of fluconazole,ketoconazole and itraconazole against clinical Malassezia isolates.Methods Broth microdilution and agar dilution methods were used to determine the minimal inhibitory concentration(MIC)of fluconazole,ketoconazole and itraconazole for 27 clinical strains(5 species)of Malassezia.Results The minimal inhibitory concentration(MIC)ranges of fluconazole,ketoconazole and itraconazole were 0.25-≥64 mg/L,≤0.03-0.5 mg/L and ≤0.03-0.125 mg/L respectively as shown by broth microdilution method,2-≥64 mg/L,≤0.03-0.5 mg/L and ≤0.03-0.25 mg/L respectively as revealed by agar dilution method.Both methods demonstrated that itraconazole possessed the strongest activity against Malassezia species,followed by ketoconazole and fluconazole.The agreement rate in MICs between the two methods was 78.8％,85.2％ and 88.9％,respectively for fluconazole,ketoconazole and itraconazole,with the intraclass correlation coefficients (ICCs)being 0.88,0.80 and 0.76 respectively.Conclusions Fluconazole,ketoconazole and itraconazole are highly active against Malassezia species in vitro,and itraconazole is the most active.Broth microdilution and agar dilution method coincide well in,and are applicable for,the antifungal susceptibility testing of Malassezia species in vitro.

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.