OBJECTIVE To explore the effect of fluconazole on proteomics of Trichosporon asahii so as to reveal the responding process of T.asahii to fluconazole stress and the resistance mechanisms to azoles on the protein level.METHODS T.asahii AS 2.2174 was chosen as the research subject,the minimum inhibitory concentration(MIC)of fluconazole was determined by broth microdilution assay.The protein abundance of T.asahii was detec-ted by means of tandem mass tag(TMT)technique combined with liquid chromatography-tandem mass spectrom-etry(LC-MS/MS)before and after the treatment with fluconazole(1× MIC).The differentially expressed pro-teins(DEPs)were identified based on the screening standards of fold change ≥1.20 or ≤0.83 and P＜0.05.Gene ontlogy(GO)and Kyoto encyclopedia of genes and genomes(KEGG)enrichment analysis were performed for the DEPs so as to understand the biological property of the DEPs and the major biological pathways that the DEPs in-volved in.Finally,the targeted validation was carried out for the targeted differentially expressed proteins by using multiple reaction monitoring(MRM).RESULTS The MIC of fluconazole to T.asahii AS 2.2174 was 8 μg/ml.Totally 196 DEPs were identified,including 93 upregulated DEPs and 103 downregulated DEPs.The function en-richment analysis showed that the DEPs mainly participated in synthesis and metabolism of sterols,drug metabo-lism,stress response,energy metabolism and intertranslation.The targeted DEPs showed the consistent expres-sion trends in MRM target validation and TMT-LC-MS/MS.CONCLUSIONS The protein abundance of T.asahii has remarkable change under the fluconazole stress.The bioinformatics analysis reveals the complicated molecular mechanisms of T.asahii in response to the fluconazole stress,which may offer valuable ideas for understanding the drug resistance to azoles and developing new drug targets.

OBJECTIVE To explore the effect of fluconazole on proteomics of Trichosporon asahii so as to reveal the responding process of T.asahii to fluconazole stress and the resistance mechanisms to azoles on the protein level.METHODS T.asahii AS 2.2174 was chosen as the research subject,the minimum inhibitory concentration(MIC)of fluconazole was determined by broth microdilution assay.The protein abundance of T.asahii was detec-ted by means of tandem mass tag(TMT)technique combined with liquid chromatography-tandem mass spectrom-etry(LC-MS/MS)before and after the treatment with fluconazole(1× MIC).The differentially expressed pro-teins(DEPs)were identified based on the screening standards of fold change ≥1.20 or ≤0.83 and P＜0.05.Gene ontlogy(GO)and Kyoto encyclopedia of genes and genomes(KEGG)enrichment analysis were performed for the DEPs so as to understand the biological property of the DEPs and the major biological pathways that the DEPs in-volved in.Finally,the targeted validation was carried out for the targeted differentially expressed proteins by using multiple reaction monitoring(MRM).RESULTS The MIC of fluconazole to T.asahii AS 2.2174 was 8 μg/ml.Totally 196 DEPs were identified,including 93 upregulated DEPs and 103 downregulated DEPs.The function en-richment analysis showed that the DEPs mainly participated in synthesis and metabolism of sterols,drug metabo-lism,stress response,energy metabolism and intertranslation.The targeted DEPs showed the consistent expres-sion trends in MRM target validation and TMT-LC-MS/MS.CONCLUSIONS The protein abundance of T.asahii has remarkable change under the fluconazole stress.The bioinformatics analysis reveals the complicated molecular mechanisms of T.asahii in response to the fluconazole stress,which may offer valuable ideas for understanding the drug resistance to azoles and developing new drug targets.

Objective:To evaluate the efficacy of medical cold patches in relieving burning pain and restoring skin homeostasis after hematoporphyrin monomethyl ether-based photodynamic therapy (HMME-PDT) for the treatment of port-wine stains.Methods:Forty patients with port-wine stains in the middle face, who met the inclusion and exclusion criteria, were collected from Department of Dermatology, the Seventh Medical Center of Chinese PLA General Hospital from November 2019 to April 2021, and randomly and equally divided into test group and control group. Patients in the test group received cold compress with medical cold patches at treatment sites for 1 hour immediately after HMME-PDT, and then once a day for 3 consecutive days, while those in the control group received no special treatment and experienced a spontaneous recovery. Pain numeric rating scale (NRS) scores were recorded immediately, 0.5, 1 and 12 hours after HMME-PDT. Skin surface temperature was measured 10 minutes before, and immediately, 30 minutes and 1 hour after HMME-PDT. Transepidermal water loss (TEWL) and water content of the stratum corneum (WCSC) were measured 10 minutes before, and immediately, 24, 48 and 72 hours after HMME-PDT. The scabbing rate was calculated at weeks 1, 2 and 3 after HMME-PDT. Two-way repeated measures analysis of variance was used for comparisons of observation indicators at different time points before and after treatment, and Bonferroni or Sidak′s test was used for comparisons between groups and within groups.Results:There were no significant differences in age, gender composition, TEWL or WCSC between the test group and control group before HMME-PDT (all P > 0.05) . Immediately after HMME-PDT, no significant difference in the NRS score was observed between the test group and control group (8.00 ± 1.17 vs. 8.20 ± 1.06, F = 0.30, P = 0.592) ; at 0.5 and 1 hour after HMME-PDT, the NRS score was significantly lower in the test group (6.25 ± 1.29, 4.80 ± 0.77, respectively) than in the control group (7.15 ± 0.99, 6.50 ± 0.69, respectively, both P < 0.05) . Immediately after HMME-PDT, the skin surface temperature in the test group and control group increased to 35.21 ± 1.333 ℃ and 35.64 ± 0.832 ℃, respectively, and there was no significant difference between the two groups ( P = 0.062) ; at 30 and 60 minutes after HMME-PDT, the skin surface temperature in the test group was 29.11 ± 1.59 ℃ and 32.46 ± 1.07 ℃ respectively, which were significantly lower than those in the control group (35.01 ± 0.91 ℃, 34.86 ± 0.74 ℃, F = 212.63, 100.20, respectively, both P < 0.001) . At 48 and 72 hours after HMME-PDT, the TEWL in the test group was 12.44 ± 0.67 g·h -1·m -2 and 10.85 ± 0.81 g·h -1·m -2 respectively, which were significantly lower than those in the control group (14.61 ± 0.34 g·h -1·m -2, 14.93 ± 0.24 g·h -1·m -2, F = 195.87, 520.54, respectively, both P < 0.001) , while the WCSC was significantly higher in the test group (57.83 ± 9.29 AU, 52.64 ± 8.09 AU, respectively) than in the control group (43.87 ± 4.82 AU, 38.68 ± 5.33 AU, F = 24.41, 49.22, respectively, both P < 0.001) . At 1 week after HMME-PDT, scab formation was observed in 3 cases in the test group, as well as in 6 cases in the control group, and there was no significant difference in the scabbing rate between the two groups ( P = 0.451) . Conclusion:The application of medical cold patches after HMME-PDT for the treatment of port-wine stains can reduce skin surface temperature, exert analgesic effects, shorten duration of postoperative pain, and promote the recovery of skin permeability barrier function.

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.

Day care practice at children′s specialist hospitals can improve the quality of care for children patients.The hospital adopted such means as improving the service management and regulations, the service process of day care, creating the " green channel" , and moving forward and extending the care service. Their practice proved that the day care mode can significantly shorten the waiting time for hospitalization, cut back medical costs, improve the medical experience of children and their family, and achieve all-win for " community-hospital-patient".

Objective To investigate the genotype and antifungal susceptibility of AspergiIlus species isolated from clinical settings and environmental sources. Methods Random amplification of polymorphic DNA (RAPD) analysis was performed to profile the genotypes of 48 Aspergillus fumigatus strains, 59 Aspergillus flavus strains and 30 Aspergillus niger strains isolated from clinical settings and environmental sources. NCCLS M38-A protocol was carried out for antifungal susceptibility testing of these Aspergillus isolates. Results As RAPD analysis showed, the 48 Aspergillus fumigatus strains were classified into 8 genotypes, 59 Aspergillus flavus strains into 12 genotypes, and 30 Aspergillus niger strains into 5 genotypes. A significant difference was observed in the susceptibility to terbinafine among the 8 genotypes of Aspergillus fumigatus (χ2 = 33.092, P ＜0.01 ) as well as in that to amphotericin B among the 5 genotypes of Aspergillus niger (χ2 = 15.185, P＜ 0.05).No statistical difference was found in the susceptibility to amphoteriein B, itraconazole, fluconazole or flucytosine among the 8 genotypes of Aspergillus fumigatus or in that to terbinafine, itraconazole, fluconazole or flucytosine among the 5 genotypes of Aspergillus niger. Conclusion There is some difference in the susceptibility to some antifungal agents between different genotypes of Aspergillus fumigatus and Aspergillus niger.

Objective To observe morphological characteristics and activity distribution of T. asahii biofilm. Methods The morphological characteristics of T. asahii biofilm were observed under an inverted microscope and scanning electron microscope, and activity was measured and quantitatively analyzed by 2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino) carbonyl]-2H-tetrazo-lium hydroxide (XTT) assay and viable count, respectively. Spatial distribution of dead/vital cells, activity and thickness of biofilm at different layers were assessed under a confocal laser scanning microscope (CLSM) following double staining with FDA/PI. Results T. asahii formed a biofilm in vitro on the surface of polystyrene materials. Under a scanning microscope, the biofilm displayed a complex three-dimensional structure which composed of spores, pseudohy-pha and true hypha. As time prolonged, the activity and quantity of biofilm increased. The results of XTT assay were correlated with those of viable count (r = 0.94, P < 0.01). The activity was of no obvious difference between different layers of the biofilm. The thickness of biofilm varied from 14.3 μm to 31 μm. Conclusions The structure of T. asahii biofilm in vitro is more complex than that of planktonic T. asahii. The activity is of no significant difference between different layers of T. asahii biofilm.

OBJECTIVE To supervise the environmental fungal load and species distribution in transplantation department and intensive care unit of the Southwest Hospital,and to analyze the relationship with season,temperature,humidity,ventilation and personnel activities.METHODS Data from Dec 2005 to Jan 2006 were collected from liver transplantation department(LTD),cerebral surgery intensive care unit(CSICU) and central intensive care unit(CICU).Air,surfaces and tap water were sampled twice a month at each department.RESULTS The air fungal load was 123.63 CFU/m3,139.90 CFU/m3,7 CFU/m3 and 217.71 CFU/m3 at LTD,CSICU,CICU and outdoor,respectively.The five most prevalent fungi collected from air and surface were Penicillium spp,Cladosporium spp,Alternaria spp,Aspergillus spp and Saccharomyces spp in turn.The five most prevalent fungi collected from water were Saccharomyces spp,Candida spp,Aspergillus spp,Penicillium spp and Rhodotorula spp in turn.The fungal load in LTD was positively correlated with the average temperature and the average humidity;the fungal load in CSICU was correlated with the average temperature and the average humidity,but the correlation between air fungal load and personnel activities wasn′t observed.CONCLUSIONS It demonstrated the fungi are found in the environment of the hospital including air,surface and water.The air fungal load varies throughout the year.The crest-time is May to June and September to October.Air fungal load is lower in winter and higher in summer and autumn.The correlation between air fungal load and temperature and humidity is observed.

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 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.