Objective To report Trichosporon asahii first isolation and identification of TH chosporon asahii in chain.Methods A clinical infection fungus strain was isolaled from mouth pseudomembrane vaginal,nasal secretions,urine,faces,and live puncture tissue,injured skin tissue of a female patient,who had no clear underlying disease.Through cultured in medium,biochemical assay,identification of the bacterial race molecular biology test and DNA sequencing.Result We demostrated that the fungus strain was unanimously identified as Trichosporon asahii.Conclution The Trichosporon asahii could induce the disseminated Trichosporonosis.

Objective To explore the feasibility of the nested PCR technique to detect the Trichosporon asahii DNA in the blood and liver samples from the murine model of disseminated trichosporonosis.Methods On day 3,7,14 after intravenous inoculation of Trichosporon asahii suspension,blood and liver samples of 30 BALB/c mice were collected.Another ten normal mice serve as control.DNA was amplified by nested PCR,with a pair of primer.Fungal culture was done as control.Results On day 3,7,14 after inoculation,the positive rate of the serum fungal cultivation were 0,20%,0;the positive rate of the serum after nested PCR assay was 80%,90%, 88.9%;the positive rate of liver fungal cultivation were 50%,40%,22.2%;the positive rate of liver after nested PCR assay was 90%,90%,88.9%.There were statistical differences between the positive rate by nested PCR assay and fungal cultivation to check the serum and liver samples on day 3,7,14 after inoculation.The survival mice to the 14~(th) day were too less to make statistical analysis.Conclusion The results suggest that T.asahii DNA assay by nested PCR is more specific,more sensitive,and faster than fungal cultivation.The nested PCR assay could be a potential assay for the diagnosis of disseminated trichosporonosis.

Objective To report the first case of disseminated trichosporonosis in China.Methods A series of clinical,histopatholog ic,and mycologic studies were carried out in a 20year-old-female patien t,who had no definite underlying disease.The causative fungus was identified according to culture,biochemical t est and DNA sequencing.Results Abundant fungal spores,yeasts and h yphae were shown in the specimens tak en from infectious granulomas.The fungal c olonies in the culture of skin lesion s or crusts,liver,oral psedomembra ne,vagina,nasal cavity,urine,and sto ol were the same.The colonies exhibi ted creamy white to yellowish in colo r,rugose on the surface,and hyphae at t he periphery.Under microscopy,there were a lot of rectangular arthrospores,round or oval spores w ith or without buddings,as well as br anched and septate filaments.The isolated fungus was unanimously identified as Trichosporon asahii(No.AS 2.2174)through culture,bio-chemistry and molecular biology tests.The patient's condition was impr oved obviously after the combinatio n treatment of amphotericin B liposom e(Amphotec)and fluconazole.Conclusion The extensive and lasting systemic impairments caused by T.asahii have scarcely been reported at home a nd abroad.Combination therapy with amphotericin B and fluconazole is effective.

Objective To investigate the role of TGF-? 1 and -? 2 in the pathogenesis of scleroderma(SD) and the relationship between this kind of cytokines and the clinical types of SD. Methods The TGF-? 1 and -? 2 mRNA and protein expressions in lesions of 17 patients with SD and 10 normal controls were detected using in situ RT-PCR technique and immunohistochemistry SP assay respectively. Results ①The positive rate and the expression strength of TGF-? 1 mRNA expression in the SD group were much higher than those in control group. There was no difference in TGF-? 1 mRNA expression between the localized SD and SSc patients. ②The positive rate and expression strength of TGF-? 1 and -? 2 proteins in SD group were much higher than those in control group. There was no difference in TGF-? 1 and -? 2 protein expressions between localized SD and SSc cases. Conclusion ①The positive rate and expression strength of TGF-? 1 mRNA in SD patients increased, which implied that TGF-? 1 mRNA may play an important role in fibrosis of SD. ②The positive rate and expression strength of TGF-? 1 and -? 2 proteins were more elevated in SD,which suggested that TGF-? 1 and -? 2 proteins were associated with skin fibrosis of SD. ③There was no relationship between the expression of TGF-? 1 and TGF-? 2 mRNA or proteins and the clinical types of SD, which indicated that there may be a similar pathogenesis for localized SD and SSc.

Objective Dentritic cell-specific ICAM-3 grabbing nonintegrin (DC-SIGN) is an important receptor for pathogenic microbes, and it acts as pathogen-recognition receptor (PAMP) and provides a bridge between the innate immunity and acquired immunity. It is meaningful to clarify whether DC-SIGN takes part in the course of cutaneous Trichosporon asahii infection and how it manages the other cytokines. Methods Twenty-five male mice were hypodermically injected with 3.2?107 CFU/ml of Trichosporon asahii suspension on one side of the back, the other side of the back received no injection to serve as control. Another five mice were treated with 0.9% saline solution as control. The cutaneous specimens were obtained on the 1st, 3rd, 7th and 14th day after inoculation. The expression of DC-SIGN gene and the other genes related to cutaneous immunity after cutaneous infection of Trichosporon asahii were investigated by RT-PCR and microarray. Results The DC-SIGN products containing 242 bp of nucleic acid from the skin specimens of the inoculated sites were amplified with RT-PCR on the 3rd, 7th and 14th day after inoculation. The results of microarray showed that a series of genes were down-regulated, including complement component 2, TNF-inducible protein cg12-1 (CG12-1), interferon-induced protein with tetratricopeptide repeats 3 (Ifit3), CD53 antigen, CD22 antigen, and prostaglandin E receptor 4. Conclusions The results suggest that DC-SIGN takes part in the course of Trichosporon asahii infection. DC-SIGN not only inhibits Th1 type of cell-mediated immunity, but also inhibits some of complement factors, prostaglandin E receptor and immunoglobulin binding protein 1b, thus resulting in chronic and intractable infection of Trichosporon asahii. The detailed roles of DC-SIGN in the course need to be further studied.

Objective To investigate the role of the ERG11 gene in the drug resistance of Trichosporon asahii (T.asahii), and to explore the relationship between the gene expression and drug concentrations. Methods Stable fluconazole-resistant strains of T.asahii were induced in vitro following exposure to a series of concentrations of fluconazole. Fluconazole-sensitive and-resistant strains of T.asahii were separately cultured in the medium containing fluconazole at concentrations of 0, 0.25, 0.5, 1, 2, 4, 8, 16, 32 and 64 μg/ml. Real-time quantitative PCR was performed to determine the mRNA expression of ERG11 gene. Results In fluconazole-free medium, the fluconazole-resistant strain of T.asahii showed significantly increased mRNA expression of the ERG11 gene compared with the fluconazole-sensitive strain (7.542 ± 5.311 vs. 1.014 ± 0.012, t=3.002, P=0.03). Additionally, the mRNA expression of ERG11 gene was also significantly higher in the fluconazole-resistant strains than the fluconazole-sensitive strains in the culture medium containing fluconazole at different concentrations of 0.25 (9.183 ± 3.226 vs. 3.281 ± 2.068), 0.5(13.657 ± 5.428 vs. 3.459 ± 1.923), 1(15.292 ± 7.007 vs. 3.242 ± 2.530), 2(13.720 ± 8.550 vs. 3.651 ± 0.728), 4(13.949 ± 2.960 vs. 3.969 ± 1.924)and 8(13.123 ± 6.429 vs. 3.824 ± 1.875)μg/ml(all P<0.05). However, no significant correlation was observed between the mRNA expression of ERG11 gene and fluconazole concentrations(fluconazole-resistant strains: rs = 0.229, P = 0.096; fluconazole-sensitive strains:rs=0.166, P=0.357). Conclusion Overexpression of ERG11 gene is associated with fluconazole resistance in T.asahii, but there is no correlation between the mRNA expression of ERG11 gene and fluconazole concentrations.

Objective To investigate the role of transforming growth factors(TGF-?1 and ?2) in the pathogenesis of scleroderma(SD). Methods The mRNA expression of TGF-?1 and ?2 in the skin lesions from 17 patients with SD and skin from 10 normal controls were detected with in situ RT-PCR technique. Results A higher positive rate and stronger expression of TGF-?1 mRNA in SD skin lesions were seen, compared with those in controls(P0.05).The higher positive rate and stronger expression of TGF-?1 mRNA than TGF-?2 mRNA in SD were seen(P

Objective To evaluate the efficacy and safety of a superpulse-mode fractional carbon dioxide(CO2) laser for the treatment of onychomycosis. Methods Patients with typical clinical manifestations of onychomycosis and positive for direct microscopic examinations of fungi were enrolled into this study, and treated with a superpulse-mode fractional CO2 laser for eight sessions. The scoring clinical index for onychomycosis (SCIO)and onychomycosis severity index (OSI)were calculated according to patients′ age, clinical type of onychomycosis, thickness of nails, area and length of nail involvement before the treatment, at the end of treatment, 1 month and 3 months after completion of treatment. Mycological clearance was also evaluated according to direct microscopy and fungal culture results. Adverse reactions to laser therapy were recorded. Statistical analysis was carried out by using the chi-square test and Wilcoxon signed-rank sum test with the SPSS 17.0 software. Results Totally, 20 patients with onychomycosis were enrolled into this study, and 75 affected nails were treated. Finally, 18 patients with 71 target nails completed the treatment and follow-up. The SCIO and OSI were 13.07 ± 6.47 and 21.11 ± 11.94 in these patients at baseline respectively, both significantly different from those at the end of treatment(9.03 ± 6.14 and 13.63 ± 12.10, respectively, both P < 0.05), 1 month((8.51 ± 6.99 and 14.18 ± 13.65, respectively, both P < 0.05)and 3 months(7.89 ± 7.26 and 13.70 ± 13.93 respectively, both P <0.05)after completion of treatment. No significant differences were observed in mycological clearance rates between the posttreatment time points(57.75%(41/71)at the end of treatment vs. 59.15%(42/71)at 1 month vs. 61.97%(44/71) at 3 months after completion of treatment, P > 0.05). The SCIO and OSI decreased from 12.48 ± 5.41 and 16.44 ± 9.89 at the baseline to 5.01 ± 5.56 and 6.44 ± 8.26 at 3 months after the treatment, respectively, in patients with distal and lateral subungual onychomycosis (DLSO), and from 17.86 ± 3.98 and 34.05 ± 2.56 to 15.88 ± 4.10 and 31.00 ± 7.28 respectively in patients with total dystrophic onychomycosis (TDO). During the treatment, several patients felt transient mild pain, but no subungual hemorrhage or other adverse reactions occurred. Conclusions The fractional CO2 laser in superpulse mode shows a reliable efficacy for the treatment of mild to moderate onychomycosis such as DLSO, especially when the nail plate is superficially invaded and grows rapidly. It directly inhibits and kills fungi, and treatment duration should be prolonged according to conditions.

Objective To summarize the progress in dermatological researches in the past 5 years achieved domestically and abroad,and venture to propose the developmental orientation and problems to be emphasized in dermatology in the next 5 years. Methods The achievements and progresses gained in dermatology in the recent 5 years were retrieved by employing the well-know informatics technology. Results Remarkable achievements have been gained in dermatological researches during recent 5 years,especially those concerning psoriasis,connective tissue diseases,vitiligo and dermatomycosis. Two professional journals in dermatology were founded by military medical service in recent 5 years. Several awards for the achievements in dermatology were rewarded,such as,one each,the first-and second-class Army Science and Technology Progress Award,second-class Army Medial Achievements Award,second-class Chinese Science and Technology Award in medicine,provincial level second-class Science and Technology Progress Award,etc. Apart from those awards,researches on investigation and prevention of skin diseases during military training had made important breakthrough. Conclusions In the next 5 years,the points of focus in dermatological research should still be put on serving the basic army units,raising the level of prevention and treatment of skin diseases in the course of training,ensuring the psychological health of soldiers,supporting the important research items and promoting further the development of dermatology.

Objective:To evaluate the effect of microevolution on phenotypes and drug resistance of the Trichosporon asahii biofilm. Methods:The standard strain of Trichosporon asahii was obtained from the Fungal Biodiversity Institute of the Royal Netherlands Academy of Arts and Sciences, the fluconazole-sensitive primary strain (TO) of Trichosporon asahii was isolated from a case of trichosporonosis diagnosed in the Department of Dermatology, the Seventh Medical Center of Chinese People′s Liberation Army General Hospital in 2000, and the fluconazole-resistant evolved strain (TEVO) of Trichosporon asahii was isolated from the above patient in 2014. Biofilms of the above-mentioned strains were formed in vitro, and tetrazolium salt XTT reduction assay was performed to evaluate growth kinetics of the Trichosporon asahii biofilm, and laser scanning confocal microscopy to determine the thickness of the biofilm; the sessile minimum inhibitory concentrations (SMICs) of fluconazole, itraconazole and voriconazole against the biofilms at different growth stages were determined in vitro for the evaluation of the resistance of the biofilms. One-way analysis of variance was used for comparisons among multiple groups, and Hartley test for testing homogeneity of variance. If the variance was homogeneous, least significant difference test was used for multiple comparisons; if the variance was heterogeneous, Tamhane′ T2 test was used for multiple comparisons. Results:In the adhesion (0 h) and formation stages (4- 24 hours) of the Trichosporon asahii biofilm, the metabolic activity of the evolved strain TEVO was the weakest (adhesion stage: F = 35.705, P < 0.001; formation stage: F = 15.042, P < 0.001) . At 48 hours after adhesion, the biofilms matured, and the TO strain showed the weakest metabolic activity ( F = 10.985, P < 0.001) . In the maturation stage, the biofilm thickness of the TEVO strain (26.1 ± 1.18 μm) was significantly higher than that of the TO strain (22.8 ± 1.73 μm, P = 0.001) , but significantly lower than that of the standard strain (29.5 ± 1.28 μm, P = 0.001) . As drug susceptibility testing showed, the SMICs of azole antifungal agents against the TEVO strain were higher than those against the TO strain in the adhesion and formation stages of the Trichosporon asahii biofilm, and the SMICs of azole antifungal agents against the biofilms of the 3 strains of Trichosporon asahii were all over 1 024 mg/L in the maturation stage of the biofilm. Conclusion:Under the dual pressure of host environment and antifungal drugs, adaptive changes took place in the phenotypes of the Trichosporon asahii biofilm with an increase in the resistance to azole antifungal drugs.