Objective:To analyze clinical features of coronavirus disease 2019 in patients with autoimmune blistering diseases (AIBDs), and to explore risk factors for the development of severe coronavirus disease 2019 in patients with AIBDs.Methods:Clinical data were collected from patients with AIBDs who were hospitalized in Zhongda Hospital, Southeast University from August 2021 to December 2022, and were retrospectively analyzed. Independent sample t-test, Mann-Whitney U test, or chi-square test (Fisher′s exact test) were used to analyze factors associated with severe coronavirus disease 2019 in AIBD patients. Results:A total of 30 patients with AIBDs accompanied by coronavirus disease 2019 were included, they were aged 62.90 ± 15.72 years, and included 16 males and 14 females; pemphigus vulgaris (12 cases, 40.00%) and bullous pemphigoid (13 cases, 43.33%) predominated in the disease spectrum of AIBDs in these patients. The common clinical manifestations of coronavirus disease 2019 were cough (25 cases, 83.33%), fatigue (22 cases, 73.33%), and fever (15 cases, 50.00%) ; most patients had mild clinical symptoms, while 7 (23.33%) developed into severe coronavirus disease 2019, including 2 out of 14 aged 30 - 59 years and 5 out of 16 elderly patients aged ≥ 60 years, and 2 out of the 5 elderly patients finally died. According to the clinical classification criteria of coronavirus disease 2019, the 30 patients were divided into the mild-to-moderate group (23 cases) and severe group (7 cases) ; the duration of AIBDs was significantly shorter in the severe group (10.71 ± 10.72 months) than in the mild-to-moderate group (27.61 ± 19.67 months, t = 2.16, P = 0.040) ; the remission rate of AIBDs was significantly higher in the mild-to-moderate group (21/23) than in the severe group (4/7, χ2 = 4.36, P = 0.037) ; the average daily dosage of methylprednisolone during the last month was significantly lower in the mild-to-moderate group ( M[ Q1, Q3]: 8 [4, 14] mg) than in the severe group (24 [12, 47] mg, U = 133.50, P = 0.007), and the proportion of patients treated with methylprednisolone at an average dosage of > 8 mg was significantly higher in the severe group (6/7) than in the mild-to-moderate group (8/23, χ2 = 5.59, P = 0.031) . Conclusions:Among the patients with AIBDs accompanied by coronavirus disease 2019, the incidence and mortality of severe coronavirus disease 2019 were higher in elderly patients than in young patients. The short course and no-remission of AIBDs, and the average daily dosage of methylprednisolone being>8 mg/d during the last month were risk factors for the development of severe coronavirus disease 2019 in the patients with AIBDs.

Objective To optimize experimental conditions for transfecting primary human melanocytes with Wnt5A-specific siRNA,and to establish the Wnt5A gene silencing model.Methods Primary human melanocytes (PHM)were cultured in vitro.Positive control GAPDH-siRNAs at different concentrations of 0,20.0,33.3,40.0 and 60.0 nmol/L were transfected into PHM by using liposomes.Realtime fluorescence-based quantitative PCR (qPCR) was performed to select the optimal concentration of siRNA with the highest transfection efficiency.Three Wnt5A-siRNAs including Wnt5A-siRNA-793,Wnt5A-siRNA-943 and Wnt5A-siRNA-1743 were constructed and transfected into PHM separately,and qPCR was conducted to select the most specific Wnt5A-siRNA.Then,the most specific Wnt5A-siRNA was transfected into PHM,and the total mRNA and total protein were extracted after 24-,48-and 72-hour treatment.qPCR and Western blot analysis were performed to determine the optimal time with the highest transfection efficiency.Results There were significant differences in the mRNA expression of GAPDH among cells transfected with GAPDH-siRNAs at different concentrations of 0,20.0,33.3,40.0 and 60.0 nmol/L (1.009 ± 0.161,0.086 ± 0.010,0.140 ± 0.016,0.285 ± 0.095,0.012 ± 0.007 respectively;F =69.469,P ＜ 0.05).Additionally,the 60-nmol/L GAPDH-siRNA group showed the lowest GAPDH mRNA expression (all P ＜ 0.05),as well as the best transfection efficiency.The mRNA expression of Wnt5A differed in the Wnt5A-siRNA-793 group,Wnt5A-siRNA-943 group,Wnt5A-siRNA-1743 group and the blank control group (0.331 ± 0.010,2.229 ± 0.029,0.078 ± 0.006 and 1.000 ± 0.024 respectively;F =7 006.094,P ＜ 0.05),and the Wnt5A-siRNA-1743 group showed the lowest Wnt5A mRNA expression (all P ＜ 0.05),as well as the best transfection efficiency.The mRNA expression of Wnt5A differed among the Wnt5A-siRNA-1743 group after 24-,48-and 72-hour treatment and the blank control group (0.396 ± 0.002,0.026 ± 0.008,0.131 ± 0.079,1.025 ± 0.276 respectively;F =29.215,P ＜ 0.05),so did the protein expression of Wnt5A (112.798 ± 0.218,77.765 ± 0.415,30.540 ± 0.219,130.025 ± 0.158 respectively;F =79 122.889,P ＜ 0.05).Moreover,the Wnt5A mRNA expression was significantly lower in the Wnt5A-siRNA-1743 group at 48 hours compared with that at 24 and 72 hours and the blank control group (all P ＜ 0.05),while the Wnt5A protein expression was significantly lower in the Wnt5A-siRNA-1743 group at 72 hours compared with that at 24 and 48 hours and the blank control group (all P ＜ 0.05).Conclusion The siRNA-mediated Wnt5A gene silencing model of human melanocytes is established successfully.

Objective:To measure the skin thickness in patients with erysipelas by high-frequency ultrasonography (HF-USG), and to compare the clinical efficacy of systemic antibiotics alone versus their combination with glucocorticoids in the treatment of erysipelas.Methods:Hospitalized patients with erysipelas were enrolled from Zhongda Hospital, Southeast University from January to December in 2021, and randomly divided into the study group and control group according to the order of visits. The study group was treated with systemic cefathiamidine for 7 days followed by oral methylprednisolone at a dose of 0.4 mg·kg -1·d -1, while the control group was treated with cefathiamidine alone. Before and after the treatment for 10 days, the thicknesses of the epidermis-dermis layers and subcutaneous tissues were measured by HF-USG at the sites of the most severe skin lesions on the affected limbs and at the corresponding sites on the healthy limbs, and white blood cell (WBC) counts, neutrophil (NEU) counts, as well as C-reaction protein (CRP) levels were determined. The t test and non-parametric test were used to compare the efficacy between two groups. Results:A total of 23 patients with erysipelas were enrolled. Among the 12 patients in the study group, 8 were males and 4 were females, and their age was 71.4 ± 11.4 years. Among the 11 patients in the control group, 7 were males and 4 were females, and their age was 67.4 ± 11.1 years. Before treatment, the thicknesses of the epidermis-dermis layers (0.33 ± 0.12 cm) and subcutaneous tissues (1.08 ± 0.49 cm) in the study group were not significantly different from those in the control group (0.25 ± 0.09 cm, 0.98 ± 0.46 cm; t = -1.83, -0.49, P = 0.081, 0.626, respectively). After the 10-day treatment, the thicknesses of the epidermis-dermis layers and subcutaneous tissues of the skin lesions on the affected limbs significantly decreased in both groups compared with those before treatment (both P < 0.05), and the decrease in the thicknesses of subcutaneous tissues was significantly stronger in the study group (0.32 ± 0.33 cm) than in the control group (0.10 ± 0.07 cm; t = 2.20, P = 0.039). Before treatment, the WBC counts ([11.16 ± 4.42] × 10 9/L), NEU counts ([8.26 ± 4.16] × 10 9/L) and CRP levels (median [ Q1, Q3]: 72.20 [19.28, 140.50] mg/L) in the study group were not significantly different from those in the control group ([10.10 ± 4.53] × 10 9/L, [7.21 ± 3.00] × 10 9/L, 34.40 [8.00, 74.20] mg/L, respectively; t or Z = 0.60, 0.71, -0.85, P = 0.578, 0.496, 0.196, respectively). After the 10-day treatment, the WBC counts, NEU counts, and CRP levels significantly decreased in both groups compared with those before treatment (all P < 0.05) . Conclusion:The combined treatment with systemic antibiotics and glucocorticoids could effectively alleviate skin inflammation, and more rapidly reduce the thicknesses of inflamed subcutaneous tissues in patients with erysipelas compared with systemic antibiotics alone.