Objective To analysis changes in species distribution of pathogenic dermatophytes from patients with tinea capitis in the Hangzhou Third Hospital between 2015 and 2017.Methods Demographic and mycological data were collected from 321 patients with confirmed tinea capitis in the Hangzhou Third Hospital between 2015 and 2017,and analyzed retrospectively.Results Of the 321 patients with tinea capitis,180 were males,and 141 were females.Their age of onset ranged from 1 month to 70 years.There were 154 (47.9％) preschool children and 13 (4.0％) adults.A total of 321 fungal strains were isolated,including 159 (49.5％) strains of Microsporum canis,110 (34.3％) strains of Trichophyton mentagrophytes,38 (11.8％) strains of Trichophyton violaceum,8 (2.5％) strains of Trichophyton tonsurans,4 (1.2％) strains of Microsporum gypseum,and 2 (0.6％) strains of Trichophyton rubrum.There was no significant difference in the distribution of fungal species (x2 =9.09,P ＞ 0.05) or the proportion of anthropophilic and non-anthropophilic dermatophytes among different age groups (x2 =1.85,P ＞ 0.05).The proportion of non-anthropophilic dermatophytes significantly differed among 2015,2016 and 2017 (x2 =23.82,P ＜ 0.01).From 2015 to 2017,the proportions of Microsporum canis were 32.5％,47.6％ and 59.6％ respectively,and the proportions of Trichophyton mentagrophytes were 53.0％,35.4％ and 23.7％ respectively.Conclusions During 2015-2017,the main population with tinea capitis in Hangzhou Third Hospital was preschool children.Moreover,the main pathogenic dermatophyte was Microsporum canis,whose proportion gradually increased year by year,followed by Trichophyton mentagrophytes,whose proportion gradually decreased year by year.

Objective To investigate the distribution and epidemiology of fungal pathogens in zoonotic dermatophytoses.Methods Seventy-four patients with dermatophytoses and 72 pets from 64 families,who were all culture positive for dermatophytes,were included in this study and classified into 64 family-based groups.Fungal culture and direct microscopic examination were carried out for species identification of fungal isolates,internal transcribed spacer (ITS) sequence analysis and random amplified polymorphic DNA (RAPD) analysis were performed for molecular identification and homology analysis.Results Dermatophyte species were consistent among the patients and pets from the same families for all the 64 family-based groups.A total of 146 fungal strains were isolated,including 93 Microsporum canis (M.canis) strains and 53 Trichophyton interdigitale (T.interdigitale) strains.M.canis was isolated from 42 (65.7％) family-based groups including 34 groups keeping cats and 8 groups keeping dogs,while T.interdigitale from 22 (34.3％) groups,including 14 groups keeping rabbits,6 groups keeping cats and 2 groups keeping dogs.There were 54 (75.0％) pets with obvious clinical symptoms (erythema,desquamation,depilation,etc),and 18 (25.0％) asymptomatic pets which were all cats.Among the 18 asymptomatic cats,14 carried M.canis,and 4 T.interdigitale.ITS sequencing and RAPD analysis revealed a high homology between the fungal pathogens in the same family-based groups.Conclusions M.canis and T.interdigitale are common species of dermatophytes in zoonotic dermatophytoses,and both of them have host specificity.Zoonotic dermatophytes can be transmitted between human and domestic animals,and attention should be paid to asymptomatic animals (carriers).

A 37-year-old male patient presented with persistent generalized itching erythema and papules for more than 1 month.The patient had received surgical treatment for type B3 thymoma in stage Ⅳ.Five months prior to the presentation,he developed myasthenia gravis.He also complained of chronic diarrhea for two years.Physical examination revealed white patches in the oral mucosa as well as scaly erythema of varying size on the face,trunk and extremities.Lamellar scales could be seen after scratching,while removal of scales could not result in the appearance of bleeding points.There was scaly hyperkeratotic erythema of palms and soles.Histopathological examination revealed psoriasis-like epidermal hyperplasia and parakeratosis with multiple dyskeratocytes,and some of the dyskeratocytes were surrounded by lymphocytes.There was a perivascular infiltration with a small number of lymphocytes.Immunohistochemical study showed positive staining for CD3,CD4 (dermis),CD8 (epidermis) and CD45RO,but negative staining for CD20,CD68 and CD30.The patient was diagnosed as thymoma-associated graftversus-host-like disease,myasthenia gravis and mucosal candidiasis.After treatment with tacrolimus and prednisone,the condition was gradually relieved.

A 27-year-old man presented with a 3-year history of persistent asymptomatic papules on the left chest and axirlary fossa. Multiple skin biopsies were performed and histopathology revealed mild acanthosis, extension of the dermal papilla, lichenoid lymphoid infiltrates in upper dermis. Some lymphoid cells migrated into the epidermis and formed Pautrier's microabscesses. Immunohistochemistry revealed that the infiltrating cells were positive for LCA, CD45RO, CD3, CD4 and CD8 (scattered), but negative for CD68 or CD30. Cutaneous laser confocal microscopy showed the shadow of scattered, oval or round, slightly refractive cells measuring 4-8 pm in diameter. A diagnosis of papular mycosis fungoides was made. The papules were softened with the lightening of lesional color after treatment with narrow-band ultraviolet B, topical fluticasone propionate cream and isotretinoin gel.

Objective To genotype Trichosporon spp. with rDNA-ITSAGSl-RFLP analysis followed by cluster analysis, and attempt to apply this method to rapid species identification of human pathogenic Trichosporon spp.. Methods Fourteen strains of Trichosporon, which belonged to 8 species, were collected. The rDNA-ITS/IGSl regions were amplified by PCR and sequenced. Simultaneously, the amplicons were digested separately with restriction enzymes, including Hae III, Hha I , Hae IH and Hha I , Hinf I , Msp I and Taq I . Results The 8 species of Trichosporon could be classified into 4 subgroups with rDNA-ITS-RFLP, while inter-species identification of all the 14 strains from 8 species of Trichosporon could be realized with rDNA-IGSl-RFLP. Also, those genotypes of T. asahii which had relative long phylogenic distance could even be discriminated with rDNA-IGSl-RFLP. Conclusion The rDNA-ITS/IGSl-RFLP analysis is expected to be used in rapid interspecific identification of genus Trichosporon.

A 16-year-old woman presented plaques on the left auricle and face over a period of 3 years. Fungal culture grew black-grey or dust velvety colony on Sabouraud's dextrose agar plate. A slide culture on potato dextrose agar plate showed conidiophores which were unbranched or occasionally loosely branched. The conidia were sympodial, zero- to two- septate, with rounded apices and truncated bases. The optimum growth temperature was 26℃ - 30℃. The fungus had the ability to liquefy glutin and hydrolyze starch. Anti-fungal susceptibility test showed the fungus was susceptible to itraconazole, terbinafine and amphoterecin B, but resistant to fluconazole. Cutaneous biopsy specimens revealed brown hyphae and budding yeast cells. The sequence of internal transcribed spacer (ITS) 1-ITS4 region of the isolate rDNA was assessed and compared against the Genebank databases. A 99% consistence was observed in the ITS sequence between clinical isolate and reference strain of Veronaea botryose Ciferri et Momtemartini. Based on the above findings, the mold was identified as Veronaea botryose Ciferri et Momtemartini. The lesions gradually subsided after 8-month treatment with oral itraconazole of 100 mg twice daily.

Objective To investigate the effect of electron transfer system on the hyphal formation of Candida albicans. Methods Candida albicans was cultured in RPMI 1640 supplemented with 10% new-born calf serum in 5% CO2 at 37 ℃ with or without the presence of inhibitors or activators of electron transfer system. Growth curve, morphology and percent of filamentation were observed for Candida albicans. MTT assay was used to assess the viability of Candida albicans. Results The solvents (chloroform and dimethyl sulfoxide) had no significant effect on the growth of and filamentation in Candida albicans. After incubation with thenoyltrifluoroacetone (TTFA) or benzhydroxamic acid for 24 hours, yeast cells of Candida albicans predominated in the culture. The growth of Candida albicans was significantly inhibited in log phase by the incubation with classic respiratory chain inhibitors such as rotenone, antimycin A, oligomycin, sodium azide, TTFA and sodium malonate, compared with the controls (all P < 0.01). Benzhydroxamic acid, an inhibitor of alternative oxidative pathway, also significantly inhibited the growth of Candida albicans in log phase (t = 10.92, P < 0.01). After incubation with rotenone, antimycin A, oligomycin, sodium azide, TTFA, sodium malonate, benzhydroxamic acid and disodium gnanylate, the percentage of filamentation in Candida albicans at 12 hours was 87.49 ± 0.52, 48.75 ± 4.44, 50.33 ± 8.50, 99.00 ± 1.00, 1.60 ± 0.53, 94.01 ± 0.99, 0.00 ± 0.00 and 92.33 ± 2.08, respectively, and the growth of Candida albicans at 7 hours was inhibited by (1.34 ± 0.15)%, (70.61 ± 1.02)%, (50.63 ± 5.38)%, (17.80 ± 7.89)%, (45.17 ± 1.27)%, (10.75 ± 3.62)%, (72.46 ± 1.14)% and -(5.96 ± 4.07)%, respectively. Conclusions Hyphal formation of Candida albicans could be suppressed by inhibitors of classic respiratory chain or alternative oxidative pathway, and is mainly regulated by alternative oxidative pathway.

Objective To analyze the clinical characteristics, mycology and therapeutics of 5 cases of cutaneous zygomycosis collected in recent 3 years. Methods A retrospective study was performed using clinical data on 5 cases of cutaneous zygomycosis collected in recent 3 years. Also, previous reports of this entity were reviewed. Results There were 1 male and 4 females among the 5 patients with cutaneous zygo-mycosis confirmed by mycology and/or pathology. The onset of age varied from 5 to 49 years, and course of disease from 7 months to 16 years. Of the 5 patients, 1 presented with superficial cutaneous zygomycosis, and the other 4 with gangrenous cutaneous zygomycosis; 3 had a history of trauma or surgery, 2 had no obvious inducements. Eruptions were located in the face of 2 patients and in the extremities of 3 patients. The isolate was identified as Rhizomucor variabilis in 3 cases, and species remained unclear in 2 cases. Four patients were treated by amphotericin B, and 1 by oral flueonazole as well as oral and injected itraconazole. Finally, 2 patients were healed, 1 was improved, 1 experienced no obvious improvement, and 1 died. Con-clusions Cutaneous zygomycosis is a rare severe devastating deep fungal infection. The first choice of drug is amphoteracin B for it. To improve the understanding of this disease may benefit the early diagnosis and therapy of it.

Objective To establish a diagnosis method for fungal infection using two-round PCR,and to evaluate its sensitivity in the detection of clinical specimens suspected to be infected with fungi.Methods A total of 29 specimens of clinical sputum and alveolar wash solution were collected from patients with suspicious fungal infection.All specimens uaderwent direct microscopy with 10%KOH,fungal culture,one-round PCR and two-round PCR.The fungal universal primer targeting ITS regions of rDNA was used in PCR.The detection rate for fungi was compared between these methods.Results The detection rate for fungi was 20.69%by direct microscopy,37.9%by fungal culture,17.2%by one-round PCR,and 48.3%by two-round PCR.More than one species of fungus were detected in 6.9%(2/29),3.4%(1/29)and 24.1%(7/29)of these specimens by fungal culture.one-round PCR and two-round PCR, respectively.There was a significant difference in the detection rate between two-and one-round PCR(x2=6.34,P＜0.05).With regard to the detection rate for more than one species of fungus,two-round PCR was significantly higher than one-round PCR and fungal culture(x2=4.09,6.30.bom P＜0.05).Conclusion Two-round PCR may help to improve the sensitivity of molecular diagnosis of fungus-infected specimens.

Objective To investigate the relationship between the clinical manifestations,prognosis and histopathological findings of mucormycosis.Methods The clinical data on and pathological findings from 7 cases of mucormycosis confirmed by fungal culture in the institute from 1989 to 2006 were analyzed retrospectively.Results There was 1 case of hinocerebral mucormycosis and 6 cases of cutaneous mucormycosis,among them,2 were mucormycotic necrotizing fasciitis (MNF).The condition of patients with rhinocerebral mucormycosis or MNF aggravated rapidly and all the 3 patients died from mucormycosis. Histopathological examination showed mixed infiltrates of inflammatory cells as well as necrosis and angioin vasion.On the contrary,the condition of the remaining 4 patients with cutanesus mucormycosis,who presented mainly with indurated erythematous patch,progressed slowly,and 2 patients were cured.Histologically,the lesions were characterized by granulbmatous infiltration with a few hyphae;no typical angioinva sion phenomenon was noted.There was no evidence of perineural invasion with hyphae in any of the 7 cases.ConclusionIn patients with mucormycosis,histopathological findings characterized by mixed infiltrates of inflammatory cells,numerous hyphae and typical angioinvasion phenomenon may herald a poor prognosis.