Unable to write in log file ../../bases/logs/gimorg/logerror.txt Search | Global Index Medicus
Show: 20 | 50 | 100
Results 1 - 2 de 2
Add filters

Year range
Chinese Journal of Dermatology ; (12): 302-309, 2019.
Article in Chinese | WPRIM | ID: wpr-745785


Objective To evaluate the inductive effect of interferon-γ(IFN-γ) combined with tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) on programmed necrosis of the human immortalized keratinocyte cell line HaCaT,and to explore its mechanisms.Methods In vitro cultured HaCaT cells were divided into several groups:negative control group receiving no treatment,IFN-γ group treated with 50 μg/L IFN-γ,TRAIL group treated with 4 μg/L TRAIL,and cytokine combination group treated with 50 μg/L IFN-γ and 4 μg/L TRAIL or zVAD combination group pretreated with 40 μmo/L zVAD for 1 hour followed by the treatment with 50 μg/L IFN-γand 4 μg/L TRAIL.After 48-hour treatment,the morphology of HaCaT cells were observed under a light microscope,methyl-thiazolyl-tetrazolium assay was performed to evaluate the inhibitory effect of the treatment on the proliferation of HaCaT cells,and double staining flow cytometry to detect the necrosis of HaCaT cells.Meanwhile,real-time fluorescence-based quantitative PCR (qPCR) was conducted to determine the mRNA expression of receptor interaction protein kinase 3 (RIP3) and mixed lineage kinase domain-like protein (MLKL),Western blot analysis to determine the expression of RIP1,RIP3,MLKL proteins and their phosphorylated forms (pRIP1,pRIP3,pMLKL),immunofluorescent staining to observe the distribution of pRIP3 and pMLKL in HaCaT cells,and the level of reactive oxygen species (ROS) in HaCaT cells in the above groups was detected by the fluorescence probe DCFH-DA.Statistical analysis was carried out with SPSS 22 software by using one-way analysis of variance (ANOVA) for comparing indices among different groups,and least significant difference (LSD)-t test for multiple comparisons.Results After 48-hour treatment,HaCaT cells in the cytokine combination group and zVAD combination group showed necrosis-like morphologic features.Methyl-thiazolyl-tetrazoliumassay revealed significant differences in the survival rate of HaCaT cells among the IFN-γgroup,TRAIL group,cytokine combination group,zVAD combination group and negative control group (73.16% ± 5.71%,81.46% ± 4.68%,72.18% ± 2.93%,69.67% ± 3.24% and 100%,respectively;F =24.34,P < 0.001).The necrosis rate of HaCaT cells was notably higher in the cytokine combination group and zVAD combination group (9.86% ± 1.31%,10.33% ± 2.16%,respectively) than in the negative control group (5.26% ± 0.91%,t =4.61,5.07,respectively,both P < 0.05).qPCR revealed that the mRNA expression of RIP3 and MLKL significantly increased in the cytokine combination group and zVAD combination group compared with the negative control group (tRIP3 =0.99,1.84,tMLKL =1.51,2.17,respectively,all P < 0.05).Western blot analysis suggested that the protein expression of RIP1,RIP3,MLKL,pRIP1,pRIP3 and pMLKL significantly increased in the cytokine combination group compared with the negative control group (all P < 0.05),and the zVAD combination group showed significantly decreased caspase 8 expression and increased expression of the above proteins compared with the cytokine combination group.Fluorescence microscopy showed that enhanced green dot-like or clump-like fluorescent spots (representing pRIP3) could be observed in the cytoplasm,and red fluorescent spots (representing pMLKL) could be seen on the cell membrane in the cytokine combination group.The average fluorescence intensity of ROS was significantly higher in the cytokine combination group than in the negative control group (t =702.00,P < 0.05).Conclusion IFN-γcombined with TRAIL can induce the programmed necrosis of HaCaT cells with increased level of ROS.

Chinese Journal of Dermatology ; (12): 439-442, 2017.
Article in Chinese | WPRIM | ID: wpr-686695


Objective To compare clinical and laboratory characteristics between systemic lupus erythematosus (SLE) patients with and without cutaneous vasculitis,and to investigate the correlation of cutaneous vasculitis with severe visceral involvement and laboratory biomarkers.Methods A total of 152 SLE patients with various skin manifestations were enrolled from Department of Dermatology of Huashan Hospital affiliated to Fudan University from July 2011 to October 2014.The clinical and laboratory data were collected and retrospectively analyzed.SLE patients with cutaneous vasculitis were divided into upper/lower extremity vasculitis group and livedo reticularis group.A logistic regression model was used to analyze the correlation between cutaneous vasculitis and various clinical and laboratory variables.Results Of 152 SLE patients,62 (41%) presented with cutaneous vasculitis,including 55 with upper/lower extremity vasculitis and 7 with livedo reticularis,and 90 (59%) did not have cutaneous vasculitis.Patients with upper/lower extremity vasculitis showed significantly younger age (30.54 ± 12.67 years vs.37.77 ± 12.17 years),and lower prevalence of aberrantly elevated 24-hour protein excretion (39.39% vs.64.00%) and serum urea level (2.08% vs.16.43%),but significantly higher percentage of females (98.18% vs.84.44%),higher proportions of patients with abnormal brain MRI (37.5% vs.12.19%),anemia (87.03% vs.70.93%) and positive antiribosomal P protein antibodies (77.77% vs.53.65%),and higher SLE disease activity index (SLEDAI) (14.71 ± 7.75 vs.10.68 ± 5.61) than those without vasculitis (all P < 0.05).The proportion of patients with decreased C3 level did not differ between patients with upper/lower extremity vasculitis and those without cutaneous vasculitis (P =0.362),but was significantly lower in the patients with livedo reticularis than in those without cutaneous vasculitis (28.57% vs.79.76%,P =0.008).However,no significant differences in the other variables were observed between patients with livedo reticularis and those without cutaneous vasculitis (all P > 0.05).Additionally,body mass index (BMI),abnormal lung function and other laboratory variables all did not differ among patients with upper/lower extremity vasculitis,patients with livedo reticularis and patients without cutaneous vasculitis (all P > 0.05).Logistic regression analysis revealed that after exclusion of potential effects of age and gender,cutaneous vasculitis was significantly positively correlated with abnormal brain MRI (OR =4.24,95% CI:1.17-16.13,P =0.028),and positive anti-ribosomal P protein antibodies (OR =3.97,95% CI:1.86-8.47,P =0.0004),but negatively correlated with abnormally elevated 24-hour protein excretion (OR =0.25,95% CI:0.09-0.69,P =0.009).Furthermore,cutaneous vasculitis showed no significant associations with abnormal serum urea level (OR =0.12,95% CI:0.01-1.06),decreased C3 level (OR =0.93,95% CI:0.38-2.28),anemia (OR =1.38,95% CI:0.56-3.40) or SLEDAI (OR =1.05,95% CI:0.98-1.14).Conclusions Cutaneous vasculitis is closely associated with central nervous system damage and emergence of anti-ribosomal P protein antibodies,so SLE patients with cutaneous vasculitis should be closely monitored for central nervous system damage.SLE patients without cutaneous vasculitis are more liable to kidney injury,so they also need to be closely monitored.