1.Construction of Gadd45a expression plasmid and its expression in human T cells
Yaping LI ; Ming ZHAO ; Qianjin LU
Journal of Central South University(Medical Sciences) 2011;36(5):398-404
Objective To construct Gadd45a expression plasmid and induce its expression in human T cells.Methods Gadd45a was amplified by reverse transcription PCR from human embryonic stem cells,and cloned into the pcDNA3.1 vector.The recombinant plasmid or blank plasmid was transfected into Jurkat cells or normal human CD4+T cells using electroporation,and the expression of Gadd45a was detected by quantitative RT-PCR and Western blot.Results Human Gadd45a expression plasmid was constructed successfully.Gadd45a was overexpressed both in Jurkat cells and normal human CD4+T cells after these cells were transfected with pcDNA3.1-Gadd45a.Conclusion The construction of Gadd45a expression plasmid and induction of Gadd45a overexpression in human T cells lay the foundation for further research on the role of Gadd45a in the epigenetic mechanism.
2.Effects of ultraviolet B (UVB) on the expression of Gadd45a gene and DNA methylation level in Jurkat cells
Yaping LI ; Ming ZHAO ; Qianjin LU
Chinese Journal of Dermatology 2010;43(9):629-632
Objective To explore the effects of UVB on the expression of Gadd45a gene and DNA methylation levels in Jurkat cells. Methods Jurkat cells were irradiated with UVB of 1.0 J/cm2 and 1.5 J/cm2 respectively, and collected at 6, 12, 24 and 48 hours after the irradiation. Real-time RT-PCR was used to detect the mRNA expression of Gadd45a gene and methylation-sensitive genes CD11a and CD70. Global methylation level was also measured by MethylAmp global DNA methylation quantification kit. Results After irradiation with UVB at 1.0 J/cm2, the mRNA level of Gadd45a increased but global methylation level decreased at 6, 12, 24 and 48 hours, and significant changes were observed at 6 and 12 hours for the level of both Gadd45a mRNA expression and global methylation (P < 0.01 or 0.05). Elevated mRNA expressions of CD11 a and CD70 were also noted in Jurkat cells after irradiation with UVB of 1.0 J/cm2, and significant elevation was observed at 12 hours (both P < 0.05 ). After irradiation with UVB of 1.5 J/cm2, there was a statistical increase in the mRNA expressions of Gadd45a, CD11 a, CD70, together with a statistical decrease in global methylation level in Jurkat cells, at 6, 12, 24 and 48 hours (P < 0.01 or 0.05). The mRNA expression of Gadd45a negatively correlated with the global level of DNA methylation in Jurkat cells (r = -0.395, P < 0.05). Conclusion UVB irradiation can upregulate the expression of Gadd45a, but downregulate the global DNA methylation level in Jurkat cells.
3.Current research on pathogenesis and treatment of atopic dermatitis
Tao HUANG ; Yunsheng LIANG ; Qianjin LU
Journal of Chinese Physician 2016;18(2):161-164
Atopic dermatitis (AD) is an allergic skin disease with a genetic predisposition.The pathogenesis is complex,including environment stimulation,epidermal barrier deficiency,and autoimmune disorders.The destruction of epidermal barrier stimulates the inflammatory response.In acute period,Th2 cells are activated to produce IL-4 and induce B lymphocytes to secrete IgE.Thus leads to degranulation of mast cells and basophils.After acute period,epidermis is thickened,accompanied with increasing expression levels of several chemokines and cytokines.In chronic phase,the cellular infiltration includes mainly Th1 and Th2 cells,and less Th17 and Th22 cells.The latter two cells together with their specific cytokines and chemokines are derived from keratinocytes and fibroblasts,which can produce tissue remodeling and fibrosis.So far,the treatment of AD contains allergens exposure avoid,anti-inflammatory,anti-infection,phototherapy,and immune therapy,etc.
4.Th17 cells and psoriasis
Lina CHEN ; Yuwen SU ; Qianjin LU
Journal of Chinese Physician 2016;18(2):165-168
Psoriasis is a common autoimmune disease and mainly affects skin,joints,or both.Psoriasis is also a chronic relapsing disorder that can cause physical and psychological burdens to patients.Currently it is widely accepted that the immune system is involved in the development of psoriasis.Th17 cells,a subtype of CD4 +T lymphocytes,are characterized by its ability to secrete proinflammatory cytokine IL-17.Recent studies indicate that Th17 cells play a predominant role in the pathogenesis of psoriasis and other immune-mediated inflammatory diseases.Moreover,targeted therapies have been developed and approved for the treatment of moderate-to-severe plaque psoriasis or psoriatic arthritis.This review summarizes the role of Th17 cells in the pathogenesis of psoriasis and several therapeutic biologics targeting this pathway in psoriasis.
5.Clinical characteristics of 1 006 Chinese patients with lupus erythematosus: results from the Lupus Erythematosus Multicenter Case-control Study in Chinese populations (LEMCSC)
Hui JIN ; Ming ZHAO ; Qianjin LU
Chinese Journal of Dermatology 2016;49(1):6-11
Objective To explore clinical characteristics of Chinese patients with lupus erythematosus (LE).Methods Data were obtained from the Lupus Erythematosus Multicenter Case-control Study in Chinese populations (LEMCSC).A unified standard was used to recruit patients and collect clinical information.The EpiData 3.1 and SPSS 18 softwares were utilized to input and analyze data respectively.Results One thousand and six patients (87.6% female) with lupus erythematosus (LE) were included in this analysis,of whom,887 (89.9% female) had systemic LE (SLE),and 119 (70.6% female) had isolated cutaneous LE (CLE).The most common involved system in SLE patients was skin (72.7%),followed by joints (69.2%),hematological system (60.8%),kidney (48.5%),serosa (18.2%),and nervous system (5.7%).The appearance of LE-specific skin manifestations was associated with an increased risk of arthritis (odds ratio [OR] =1.612,95% confidence interval [CI]:1.181-2.200),but with a decreased risk of nephritis (OR =0.218,95% CI:0.157-0.303) and serositis (OR =0.311,95% CI:0.218-0.443).The presence of acute CLE (ACLE) lesions was a risk factor for systemic involvement (OR =4.931,95% CI:3.232-7.524),while that of chronic CLE (CCLE) lesions was a protective factor for systemic involvement (OR =0.355,95% CI:0.234-0.541).The appearance of LE-nonspecific skin manifestations was closely correlated with the involvement of internal organs in patients with LE.Conclusion This study revealed main characteristics of LE patients in China and the relationship between LE-related skin lesions and internal organ involvement.
6.Detection of methylation in exon 1 of the survivin gene in condyloma acuminatum (CA) tissue and its significance
Qiuping ZHONG ; Hongwen ZHANG ; Fei GAO ; Qianjin LU ; Ying ZHOU
Chinese Journal of Dermatology 2011;44(5):306-309
Objective To investigate the relationship between the expression of the survivin gene and CpC methylation in exon 1 of the survivin gene in CA tissue, and to study the expression of survivin protein in CA tissue and its modulation mechanism. Methods Tissue samples were obtained from the CA lesions of 30 patients, normal cervix of 10 female controls, and normal foreskin of 10 male controls. Immunohistochemistry was carried out to detect the expression of survivin protein in these specimens, RT-PCR to measure the mRNA expression of survivin, and methylation specific PCR (MSP) to analyze the methylation status of CpG island in the survivin gene exon 1. Results The positivity rate of survivin protein and mRNA was 90% (27/30) and 93.3% (28/30) in CA tissue specimens, respectively, 5% (1/20) and 5% (1/20) in control tissue specimens, respectively; there was a significant difference between the two groups of specimens in both the parameters (x2 = 35.187, 38.437, both P < 0.01). The demethylation of CpG island in the survivin gene exon 1 was observed in 86.7% (26/30) of the CA tissue specimens and in 15% (3/20) of the control tissue specimens (x2 = 10.865, P < 0.01). There was a positive correlation between the demethylation status of CpG island in exon 1 and the mRNA expression of survivin gene (x2 = 13.929, P = 0.014). Conclusions The expression of survivin protein in CA tissues might be associated with the demethylation of CpG island in exon 1 of the survivin gene, and may play a certain role in the development of CA.
7.DNA methylation status of miR-126 and its host gene EGFL7 in CD4+T cells from patients with systemic lupus erythematosus
Yunsheng LIANG ; Sha ZHAO ; Gongping LIANG ; Ming ZHAO ; Qianjin LU
Journal of Central South University(Medical Sciences) 2013;38(8):793-797
Objective:To explore the mechanisms by which DNA methylation regulates miR-126 and its host gene EGFL7 in CD4+T cells from patients with systemic lupus erythematosus (SLE).
Methods:We analyzed the expression and the DNA methylation status within promoter region of EGFL7 and miR-126 by real-time qPCR and bisulifte genomic sequencing analysis.
Results:miR-126 and EGFL7 mRNA expression was upregulated in CD4+T cells from SLE compared with that from healthy controls (P<0.01). EGFL7 mRNA level was positively correlated with miR-126 expression in CD4+T cells from SLE (r=0.538, P=0.015). The average methylation level of EGFL7 promoter in CD4+T cells from SLE was lower than that from healthy controls (P<0.05).
Conclusion:hTe upregulation of miR-126 and its host gene EGFL7 expression in CD4+T cells from SLE is associated with the hypomethylation of the EGFL7 promoter.
8.Clinical significance of detecting food allergic specific IgG in serum of patients with eczema and chronic urticaria
Lina TAN ; Guiying ZHANG ; Lanxiang YAN ; Sheng WANG ; Qianjin LU
Journal of Chinese Physician 2010;12(4):466-469
Objective To investigate the levels of 14 kinds of food allergic specific IgG antibody in eczema and chronic urticaria patients'serum and the role of food allergens in allergic skin diseases and provide scientific basis for clinical treatment and prophylaxis. Methods We used ELISA method to detect the concentration of 14 kinds of food allergic specific IgG antibody in the serum of 900 eczema and chronic urticaria patients and 18 healthy people. Results The food allergen of positive rate in studied group and control group was 90.2% and 11.1%. The positive rate of 14 kinds of food allergic specific IgG antibody in 900 eczema and chronic urticaria patients' serum from high to low was eggs, wheat, shrimp, milk, soja,crab, rice, corn, tomato, chicken, mushroom, beef, pork and ling. The food allergen positive rate in eczema and chronic urticaria patients'serum was 91.9% and 88. 9% ( P >0. 05). The positive rate of shrimp,mushroom and ling in eczema patients'serum was higher than that in chronic urticaria patients'serum ( P <0. 05 ). The positive rate in children's group was 96. 6%, which was higher than other age groups ( P >0. 05 ). The main allergens were egg, wheat and milk in children's group. The sensitization of crab and shrimp increased as people grew up. In 812 positive patients, who were intolerant of three and more than three foods accounted for 53.7%. In 175 follow-up patients, the effective rate was 50.9% in 20 days and 61.1% in 60 days after they avoided the intolerant food. The patients'diet was adjusted according to the test results.In 325 follow-up patients, the effective rate in eczema group was 59.9% in 20 days and 70. 4% in 60 days ( P <0. 05). Thee effective rate in chronic urticaria group was 65.6% in 20 days and 77. 3% in 60 days ( P <0.05 ). Conclusion The intolerant food can be found by detecting serum food allergen specific IgG.It can be used to adjust the diet and relieve the symptom, which is of great importance for preventing and treating allergic skin diseases.
9.RUNX3 gene methylation in a cutaneous malignant melanoma cell line A375
Li ZENG ; Yuwen SU ; Shengbo YANG ; Qianjin LU ; Jing ZHOU
Chinese Journal of Dermatology 2010;43(3):199-201
Objective To investigate the relationship between the methylation of CpG island of RUNX3 gene promoter and its expression in a human cutaneous malignant melanoma cell line A375, and to assess the role of RUNX3 gene methylation in the pathogenesis of human cutaneous malignant melanoma. Methods Cultured A375 cells were treated with various concentrations (0, 1, 5, 10, 20 μmol/l) of 5-azacyti-dine for 24 or 72 hours followed by another 5 days of culture. Then, methylation-specific PCR (MSP) was performed to evaluate the methylation status of RUNX3 promoter region, and Western-blot analysis to detect the protein expression of RUNX3 in A375 cells. Results The RUNX3 gene promoter region was hypermethylated in untreated A375 cells, along with the absence of protein expression of RUNX3. However, after the treatment with 5-azacytidine, the promoter region of RUNX3 gene was demethylated partly, and the expression of RUNX3 protein was restored in A375 cells. Further, the expression intensity was directly correlated with the concentration of 5-azacytidine. Conclusions The promoter hypermethylation of RUNX3 gene may be related to the silencing of RUNX3 gene expression in A375 cells, whereas 5-azacytidine can cause the demethylation of RUNX3 gene, reactivate the gene which has been inactivated by the promoter hypermethylation, and finally induce the re-expression of RUNX3 protein.
10.Diagnosis and treatment of adult-onset Still′s disease
Xiaoqing YI ; Shuaihantian LUO ; Guiying ZHANG ; Qianjin LU
Chinese Journal of Dermatology 2021;54(2):165-169
Adult-onset Still′s disease (AOSD) is a type of systemic inflammatory disease of unknown etiology, with diverse clinical manifestations, and there are some difficulties in its diagnosis and treatment. In recent years, it has been found that some new markers, such as heme oxygenase 1, calreticulin, inflammatory cytokines and advanced glycation end products, can be used for a comprehensive assessment of the activity and severity of AOSD. Moreover, new biological agents, such as tumor necrosis factor inhibitors, interleukin-1 (IL-1) inhibitors, IL-6 inhibitors and recombinant IL-18 binding proteins, bring new hope for the treatment of AOSD. This review mainly summarizes progress in the diagnosis and treatment of AOSD.