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Objective:To analyze measurement results of serum allergen-specific immunoglobulin E (IgE) in patients with eczema/dermatitis.Methods:A retrospective analysis was conducted on serum allergen-specific immunoglobulin E (IgE) levels in 3 051 patients with eczema/dermatitis, who visited the allergy clinic of Huashan Hospital from April 1, 2021 to March 31, 2022. The serum allergen-specific IgE level was detected by using the Phadia allergen detection system, and positive rates of allergens were calculated to determine common inhaled allergens and food allergens in patients with eczema/dermatitis. Comparisons of enumeration data between groups were performed by chi-square test.Results:Among the 3 051 patients with eczema/dermatitis, there were 1 412 with atopic dermatitis and 1 639 were other eczema/dermatitis. Detection of serum allergen-specific IgE showed that 1 629 (53%) patients were positive for allergens, and the number of positive allergen-specific IgEs in each patient was 3.0 ± 1.6. The top 3 common inhaled allergens in patients with eczema/dermatitis were Dermatophagoides farinae (904/1 522, 59%) , Dermatophagoides pteronyssinus (891/1 513, 59%) and Alternaria alternata (206/1 068, 19%) , and the top 3 common food allergens were shrimps (251/1 432, 18%) , egg white (165/992, 17%) and cow milk (149/994, 15%) . Among the 3 051 patients, 25 (1%) were aged < 2 years, 571 (19%) aged 2 - 12 years, 285 (9%) aged 12 - 18 years, and 2 170 (71%) were aged > 18 years. The most common food allergens were both egg white in the age groups of < 2 years and 2 -12 years (77%, 37%, respectively) , and were both shrimps in the age groups of 12 - 18 years and > 18 years (31%, 17%, respectively) . Dermatophagoides pteronyssinus and Dermatophagoides farina were the top 2 common inhaled allergens in all age groups, with the positive rate ranging from 36% to 84%; in addition, the positive rate of molds was relatively high in the age group of 2 - 12 years (mold mixture: 37%; Alternaria alternata: 27%) . From April 2021 to March 2022, the positive rate of outdoor allergens ranged from 10% to 15% among outpatients in every month; the positive rates of tree pollen and grass pollen increased from April 2021, and peaked in October 2021. The patients with atopic dermatitis showed a significantly increased positive rate of allergens (73%) compared with those with other eczema/dermatitis (37%, χ2 = 389.36, P<0.001) , and the rank of common allergens in the patients with atopic dermatitis was basically the same as that in those with eczema/dermatitis. Conclusions:The common allergens were Dermatophagoides farina, Dermatophagoides pteronyssinus and Alternaria alternata in the patients with eczema/dermatitis. Food allergy was more common in infant patients, and inhalation allergy was more common in child, adolescent and adult patients. The positive rate of allergen-specific IgEs was markedly higher in the patients with atopic dermatitis than in those with other eczema/dermatitis.
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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.
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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.