Objective To investigate the effect of EGCG on the radiosensitivity of human nasopharyngeal carcinoma CNE-1 cells.Methods CNE-1 cells were divided into four groups:control,EGCG treatment,UVC or X-ray exposure,and EGCG combined with UVC or X-rays.After treatment with different concentrations of EGCG for 24,48 and 72 h and UVC or X-rays,cell growth was determined with MTT assay,cell survival was measured with clonogenic assay,cell cycle was deteeted with flow cytometry,cell apoptosis was detected by the annexin V-FITC cell apoptosis kit,and protein expression was assayed by Westem blot.Results EGCG inhibited cell growth in a dose-and time-dependent manner(r =0.817and 0.364).Compared with UVC or X-ray irradiation alone,the radiosensitivity of CNE-1 cells was enhanced by 2 h pre-treatment of 50 μmol/L EGCG,which disrupted S phase arrest caused by UVC( t =18.68,P ＜ 0.05 ) and increased the population of S and G2/M arrest caused by X-rays ( t =7.11 and 6.99,P ＜0.05 ).UVC could cause a significant increase of sub-G1 population( t =6.67,P ＜ 0.05 ) and Annexin V-FITC assay indicated apoptosis was further elevated by EGCG ( t =10.28,P ＜ 0.05 ).However,no significant induction of apoptosis was observed in the cells either irradiated with X-rays alone or combinationly treated with EGCG and X-rays.The combination treatment of EGCG and UVC significantly increased the expression of Bax and Caspase-3 proteins,but failed to affect Bcl-2 protein expression.Conclusions EGCG enhances the growth inhibition of CNE-1 cells caused by UVC or X-rays,which is relevant to apoptosis induction or cell cycle arrest.

Objective To compare broth microdilution and agar dilution methods for in vitro testing of activities of fluconazole,ketoconazole and itraconazole against clinical Malassezia isolates.Methods Broth microdilution and agar dilution methods were used to determine the minimal inhibitory concentration(MIC)of fluconazole,ketoconazole and itraconazole for 27 clinical strains(5 species)of Malassezia.Results The minimal inhibitory concentration(MIC)ranges of fluconazole,ketoconazole and itraconazole were 0.25-≥64 mg/L,≤0.03-0.5 mg/L and ≤0.03-0.125 mg/L respectively as shown by broth microdilution method,2-≥64 mg/L,≤0.03-0.5 mg/L and ≤0.03-0.25 mg/L respectively as revealed by agar dilution method.Both methods demonstrated that itraconazole possessed the strongest activity against Malassezia species,followed by ketoconazole and fluconazole.The agreement rate in MICs between the two methods was 78.8％,85.2％ and 88.9％,respectively for fluconazole,ketoconazole and itraconazole,with the intraclass correlation coefficients (ICCs)being 0.88,0.80 and 0.76 respectively.Conclusions Fluconazole,ketoconazole and itraconazole are highly active against Malassezia species in vitro,and itraconazole is the most active.Broth microdilution and agar dilution method coincide well in,and are applicable for,the antifungal susceptibility testing of Malassezia species in vitro.

Objective To explore the influence of honokiol on growth,cell cycle and radiosensitivity in nasopharyngeal carcinoma cells,CNE-1 and CNE-2.Methods MTT assay and clonogenic assay were used to detect cell growth and survival respectively.Flow cytometry was employed to analyze cell cycle progression.Annexin V-FITC kit was used to detect cell apoptosis.Western blot assay was applied to examine protein expression.Results Honokiol signficantly inhibited proliferation of CNE-1 and CNE-2 cells in a dose and time dependent manner,the IC50 value was 2.84 and 2.68 μmol/L(24 h)and 2.50 and 2.20 μmol/L (48 h),respectively.After being treated with 2.5 μmol/L honokiol for 24 h,the ratios of early apoptosis,late apoptosis and necrosis were 24.53％,23.05％ and 7.13％ in CNE-1 cells compared with the control group(t =-41.17,-8.18,-6.08,P ＜0.05).The expression levels of pro-apoptotic proteins Caspase-3 and Bax were significantly increased to 2.31 and 1.89 times (t =-15.92,-17.15,P ＜ 0.05),4.43 and 1.85 times (t =-29.39,-13.47,P ＜ 0.05).Simultaneously the expression level of anti-apoptotic protein Bcl-2 was reduced by 2.22 and 2.74 times(t =26.94,66.14,P ＜ 0.05) as compared with controls after being treated with 4 and 3 μmol/L honokiol.Additionally,honokiol at lower doses signicantly enhanced the senstivity of CNE-1 and CNE-2 cells to X-ray irradiation.The SER was 1.41 and 1.88 in CNE-1 and CNE-2 cells.3 Gy irradiation of X-rays increased the proportion at G2/M state in both cell lines (t =-14.96,-19.26,P ＜ 0.05).Honokiol reduced the G2/M cell cycle arrest induced by irradiation significantly (t =7.65,4.98,P ＜ 0.05).Simultaneously,cyclin B1 protein expression obviously elevated (t =-33.07,-73.49,P ＜ 0.05).Conclusions Honokiol is a potent inhibitor of nasopharyngeal carcinoma cell growth by inducing cell apoptosis and necrosis and works as a radiosensitizer by disrupting G2/M cell cycle checkpoint.

ObjectiveTo investigate the predisposing factors and pathogenic fungal species of Malassezia folliculitis in different geographical areas and body sites.MethodsTotally,241 patients diagnosed with Malassezia folliculitis were asked to complete a questionnaire.The content of hair follicles was obtained and subjected to fungal smear and culture examination.Fungal species were identified according to morphological,physiological and biochemical features.Results Of the 241 patients with Malassezia folliculitis,204 (84.65％) were positive for smear examination.A total of 259 specimens were collected from these patients,and fungal culture grew 213(82.24％) strains,of which,209 belonged to Malassezia species,4(1.54％) to Candida species.Among the 209 Malassezia strains,186 were activated and subjected to species identification which resulted in 6 species,including M.furfur (111 strains,59.68％ ),M.sloofiae (43 strains,23.12％ ),M.sympodialis (17 strains,9.14％),M.globosa (9 strains,4.84％),M.pachydermatis (4 strains,2.15％),and M.obtuse(2 strains,1.08％ ).Of the pathogenic fungi of Malassezia folliculitis,M.furfur predominated in the chest,back,abdomen,face and neck,M.sloofiae in the upper limbs,shoulders and vertex,M.globosa in the lower limbs.There were obvious differences in the distribution of pathogenic fungal species at different body sites in a same host,and M.furfur with M.sloofiae or M.sympodialis appeared to be the most common pathogens.ConclusionsIn this study,6 Malassezia species are identified in patients with Malassezia folliculitis in Nantong and Nanjing area,M.furfur and M.sloofiae appear to be the dominant pathogens.

Objective To explore the difference between T cells in the synovial fluid and peripheral blood in patients with rheumatoid arthritis(RA). Method Samples from 22 patients were studied. The differentiation and activation markers expressed on T cell surface were detected by immunofluorscence using flow cytometer. The specific proliferation of collagen Ⅱ and heat shock protein 70 was analyzed using standard 3H-TdR incorporation method. Restricted V beta usage of these T cell was analyzed by semi-quantitied RT-PCR. Results The majority of the T cell subsets in the synovial fluid were demonstrated to be CD4 and CD8 positive cells in which (40?10)% were CD4 positive and (36?16)% were CD8 T cells respectively. The ratio between CD4 and CD8 was much lower than that found in the PBL of RA patients. The percentage of CD3+/CD25+ T cells was (16?6)%. The specific proliferation of collagen Ⅱ and HSP70 to CD3+/CD25+ T cell was higher than that of CD3+/CD25+ negative T cells. The T cell receptor expressed on the T cells from both peripheral blood and synovial fluid were tested for ?? TCR (70?26)%. However, the T cells in the synovial fluid showed V?14,16 and 17 restriction. Conclusion The data here reported indicates that T cell subsets in the synovial fluid and peripheral blood circulation in patients with rheumatoid arthritis are different. The T cells in the synovial fluid demonstrates more activation and higher reactivation to collagen Ⅱ and HSP70. The TCR of T cells showes V?14,16 and 17 restriction.

Four hundred and four male patients with primary gout were enrolled. According to the degree of nonalcoholic fatty liver diseases (NAFLD), the patients were divided into simple gout ( n=121), gout combined with mild NAFLD ( n=149) and gout combined with moderate-severe NAFLD ( n=134). The height, weight, waist, hip, blood pressure and blood biochemistry parameters of patients were measured. The degree of NAFLD was negatively correlated with the age of patients in three groups. The BMI, ratio of waist/hip, count of red cells, hemoglobin, hematocrit, red blood cell distribution width ( SD and CV), triglyceride, alanine aminotransferase and HOMA-IR were increased with the increasing of NAFLD severity (all P<0.05). Red blood cell count, hemoglobin, alanine aminotransferase, serum uric acid increased with the increasing of NAFLD severity (all P<0.05). Platelet, serum urea nitrogen and serum creatinine were decreased with the increase of NAFLD severity. Logistic regression showed that BMI, hemoglobin and HOMA-IR were independent risk factors for NAFLD. The prevalence and the severity of NAFLD was increased with increasing quadrates of hemoglobin. Taking group Q1 as a control, OR of NAFLD in group Q2 was 1.166(95 %CI:0.638-2.133), OR in group Q3 was 2.011(95 %CI:1.122-3.605)and OR in group Q4 was 3.120(95 %CI:1.613-6.034). The result indicates that hemoglobin levels are associated with the development and the severity of NAFLD in male patients with primary gout.