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.

Recently, biochemical and genomic studies have specified new classification methods and renamed Propionibacterium acnes as Cutibacterium acnes ( C. acnes) to better study its phylotypes and distinguish it from other Propionibacterium species . C. acnes, an important commensal bacterium in human skin, is involved in maintaining skin health, and can also turn into an opportunistic pathogen causing acne vulgaris. Latest studies have showed that the balance between different phylotypes of C. acnes and its interaction with other microorganisms play a key role in the occurrence and development of acne vulgaris. This review summarizes correlations between C. acnes phylotypes and acne vulgaris, as well as antimicrobial susceptibility and interaction with other microorganism of C. acnes.

Objective To investigate the effect of intensive follow-up on the correction of mild anemia among infants in the community. Methods Eighty four infants aged 6-12 months with mild anemia managed systematically in the child health clinic of Changzhou Lanling Community Health Service Center were enrolled from April 2016 to January 2017. Seventy five cases completed the study, including 36 cases managed with intensive follow-up (study group) and 39 cases managed with routine procedure (control group). The hemoglobin(Hb) levels, Hb changes, the rates of correction and closure of anemia and the medication of iron treatment were compared between two groups during the 2 and 6 month-follow-up. Results The Hb value, Hb changes, the rate of correction in study group were significantly higher than those in control group after 2 monthsof treatment [(113.50 ± 5.40)g/L vs. (109.13 ± 7.91)g/L, t=2.772, P=0.007;(13.22 ± 4.56)g/L vs. (7.54 ± 6.75)g/L, t=4.240, P<0.001;77.8％(28/36) vs. 46.2％(18/39),χ2=6.617, P=0.010, respectively]. In 6 month-follow-up, the Hb value, the rate of correction and closure rate in study group were significantly higher than those in control group [(116.22±3.72)g/L vs. (112.23±5.90)g/L, t=3.471, P=0.001;88.9％(32/36) vs. 66.7％(26/39),χ2=4.082, P=0.043;77.8％(28/36) vs. 41.0％(16/39),χ2=8.967, P=0.003, respectively]. For infants receiving iron treatment, the proportion of standardized medication in study group was significantly higher than that in control group after 2 months (14/14 vs. 4/8, χ2=5.525, P=0.019); the Hb values in study group were significantly higher than those in control group after 2 months[ (109.93 ± 3.51)g/L vs. (102.88 ± 2.88)g/L, t=4.820, P<0.001)and 6 months[(114.50±4.00)g/L vs. (108.55±5.04)g/L, t=3.297, P=0.003] of follow-up. Conclusion The intensive follow-up management can significantly improve the Hb values, the rates of correction and closure rate of infants with mild anemia in the community.

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 differences in the expression of Ras 1,Rac1 and Rho1 genes between yeast and hyphal phases of Trichosporon asahii (T.asahii),and to explore their roles in the formation of hyphae.Methods The yeast phase and hyphal phase of T.asahii were cultured and served as yeast phase group and hyphal phase group respectively.Total RNA was extracted from the 2 groups,and real -time fluorescence-based quantitative PCR (RT-PCR) was performed to measure the mRNA expression of Ras1,Rac1 and Rho1.Results The hyphal formation rate was significantly lower in the yeast phase group than in the hyphal phase group (0.40％ ± 0.53％ vs.99.33％ ± 0.57％,t =13.93,P ＜ 0.05).When the mRNA expression of Ras1,Rac1 and Rho1 in the yeast phase group was all set as 1,that in the hyphal phase group was 25.17 ± 10.99,16.81 ± 7.80,42.61 ± 18.50,respectively,with significant differences between the two groups in the three parameters (t =3.81,3.51,3.90,respectively,all P ＜ 0.05).Conclusion Ras1,Rac1 and Rho1 genes may participate in the regulation of hyphal formation in T.asahii.

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.

Bibliometrics ; China ; Humans ; Periodicals as Topic