Objective To investigate characteristics of endometriotic lesions clone with various pathological subtypes by analyzing the polymorphism of X chromosome linked human androgen receptor allele (HUMARA). Methods Eight frozen tissues of endometriotic leisons were collected from 6 patients who received laparotomy or laparoscopy surgery in Peking Union Medical College Hospital from Nov. 2008 to Jun. 2009. Fifty specimens of epithelial cells from single endometrial glands were isolated and collected from endometriotic lesions by using laser capture microdissection. HUMARA was applied as the gene marker of clonal analysis. Nested palymerase chain reaction, double-enzyme digestion reaction with two methylationsensitive restriction endonuclease (Hha Ⅰ and Hpa Ⅱ), and the automated gene sequencing technique were utilized in this study to evaluate the characteristics of endometriotic lesions clone. Results Of 50 specimens of isolated glands, 34 were informative for clonal analysis, and all of which showed monoclonality. Of 3 ovarian endometriotic tissues, one tissue of HUMARA showed unuseful information, the other 2 ovarian endometriotic tissues respectively had 4 and 7 informative specimens of gland epithelial cells, and all of the glands from each tissue showed uniform clonal pattern. Two peritoneal endometriotic tissues had 1 and 5 informative specimens from individual glands, respectively; and the clonal patterns in 5 glands from the single lesion were divergent. Two abdominal wall endometriotic tissues had 7 and 6 informative specimens,respectively; and variable clonal patterns were seen in different glands from each lesion. One deep infiltrating endometriotic lesion had 4 informative specimens of isolated glands, and all of them showed unique clonal pattern. The disparate clonal patterns were found in endometriotic lesions with variable pathological subtypes, even arising from the same patient. Conclusions The epithelial cells from individual endometriotic gland showed monoclonality , and different glands from the same endometriotic lesion might show divergent patterns.

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 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 investigate the pathogenic factors and the visceral involvement in murine disseminated trichosporonosis caused by Trichosporon asahii. Methods Forty-five mice were immunosuppressed with cyclophospamide 3 days hefore and 7 days after inoculation of T. asahii, and were divided into intravenously inoculated group (n = 15), intradermal inoculated group (n = 7), gastrointestinal infusion group (n = 8), intravenously inoculated + treatment group (n = 15). In the control groups the mice were not immunosuppressed, and were also divided into intravenous, intradermal, and G.I. infusion groups with the same number of mice respectively. In the treatment group the mice were given both liposomal amphotericin B and fluconazole. The main viscera of the mice were examined by mycologic culture and pathologic sections. Results In the intravenous inoculation group of immunized mice, Trichosporon asahii were isolated from at least one organ in 10/12 mice, while T. asahii were only isolated in 2/14 mice in the control group; in 2/7 mice of the intradermal group of immunosuppressed mice, skin lesion appeared at the inoculation site, but no visceral infection was observed. No visceral infection was found in the groups that T. asahii was inoculated by non-intravenous injection in both immunosuppressed and non-immunosuppressed mice. The number of mice died, the number of visceral organs involved and the incidence of systemic infection were significantly less in the treatment group than those in the non-treatment groups (P

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.