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 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 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.

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 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.

Objective. Report of first case of Protothecosis zopfii in China and causes the skin infection in the world.Method. By clinical and laboratory examinations to confirm the diagnosis and the response to treatment. By the review of literatures to confirm the first case of human skin infection in the world.Result.From the literatures and the clinical pictures, it is confirmed that this is the first case report of Protothecosis zopfii of skin in the world.Conclusion.The first case of Protothecosis zopfii in human being was reported and successfully treated with local infiltration of Diflucan (fluconazole) 3ml (2mg/ml)/week×4.

Objective To evaluate the cell-mediated immune tissue reactions of mice model of chromoblastomycosis. Methods First we developed a murine model of chromoblastomycosis subcutaneous infection with F. pedrosoi inoculated into the footpads using immunocompetent and immunocompromised mice immuned by CTX, and then by immunohistochemistry methods analyzed the expression of cytokines IL-4, IL-10, TNF-α and IFN-γ at the seventh and thirtieth day, the expression of these cytokines at the same time in the healthy mice footpads were used as control. Results In the immunocompetent mice, The expression of IL-4, IL-10 and TNF-α was significantly higher when compared with healthy control at the seventh day,showing an up-regulation pattern of Th2 and Th1 type cellular immune functions with a predominance of the Th2 response. At the 30th day, the expression of IL-10 was significantly lower when compared with the 7th day and no difference with healthy controls, while IFN-γand TNF-α were gradually increased, the T cellmediated immune drives to Th1 response from Th2. In the immunocompromised mice, the expression of IL4 and IL-10 were significantly higher, meanwhile IFN-γwas lower than those in immunocompetent or healty mice, the levels of TNF-α was not significantly different fiom healty control at the 7th day, it showed that Th2 response was more increased with the Th1 responses was significantly inhibited in the early immune reactions. At the 30th day, the Th1 type cytokines IFN-γ and TNF-α were highly significantly higher than early stage but still lower than immunocompetent levels, the level of Th2 type cytokine IL-10 rradually decreased although it was still above the other two rroups. Conclusion In different immune state, there was an immune defence translation from the predominance of Th2 type cellular immunity to Th1 type in the process of murine model of chromoblastomycosis. Thl type cytokines which favors resistance to fungal disease, played a major role at controlling the development of chromoblastomycosis.

Objective To establish a method of gene mutation detection for congenital adrenal hyperplasia (CAH) by using sequencing, single nucleotide polymorphisms (SNP) analysis and T-A cloning. Methods The blood samples of 33 patients with 21-hydroxylase deficiency (21-OHD) , 2 patients with 17α-hydroxylase deficiency (17-OHD) , the parents of all the patients and 105 healthy children were collected. Genomic DNA were extracted form the blood samples. To detect the gene mutation of CYP21A2,highly specific primers for CYP21A2 gene were designed according to the sequence differences between CYP21A2 gene and its pseudogene. The whole CYP21A2 gene was amplified and sequenced. SNP analysis and TA cloning of PCR products were also carried out. The molecular diagnosis of 17-OHD was based on the amplification and sequencing of CYP17A1 gene. Results The corresponding gene mutations was determined in all the patients based on the method established in this study. Thirteen mutations of CYP21A2 gene were identified in 33 patients with 21-OHD. The 3 most frequent mutation of CYP21A2 gene were IVS2-13A/C >G, p. I172N and chimeric mutation, which accounted for 32% (21/66) ,27% (18/66) and 15% (10/66) respectively. Ninety-one persent mutations of CYP21A2 gene resulted from pseudogene conversion. In 2 patients with 17-OHD, homozygous mutations of CYP17A1 gene, IVS4-6A > G and p. 487_489del were identified separately. All the gene mutations detected in the patients were inherited from their parents. No mutation of CYP21A2 gene or CYP17A1 gene was found in the healthy children. Conclusion A method of gene mutation detection for CAH has been established. It will be beneficial to clinical diagnosis of CAH.

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.