1.Proliferation of desmoglein 3-stimulated T lymphocytes from patients with pemphigus vulgaris
Jie HU ; Zhongxiang WEI ; Yi JIANG ; Zaifu LIANG ; Long GENG
Chinese Journal of Dermatology 2010;43(7):474-477
Objective To investigate the impact of desmoglein 3 (Dsg3) on the proliferation of peripheral T lymphocytes from patients with pemphigus vulgaris (PV).Methods Peripheral blood mononuclear cells (PBMCs) were obtained from 12 patients with PV and 22 normal human controls,cultured with or without the presence of Dsg3 or phytohemagglutinin for 3 days.Flow cytometry was performed to detect the changes of T-lymphocyte subsets in PBMCs stimulated with Dsg3 and proliferation of T-lymphocytes.Results In patients with PV,the percentage of Th2 and Th1 cells was 12.17%±5.32% and 4.08%±1.50%,respectively in Dsg3stimulated PBMCs,9.84%±5.41% and 3.91%±1.38%,respectively in non-stimulated PBMCs.Increased percentage of Th2 cells was observed in Dsg3-stimulated and non-stimulated PBMCs from patients with PV compared with those from normal human controls (both P<0.05).After stimulation with Dsg3,there was a significant proliferation of T cells from patients,and the proliferation rate of CD4+T cells was 4.65%±3.28%,which was significantly higher than that from normal controls(P<0.05).Conclusion Dsg3 can induce the specific proliferation of CD4+T cells,especially Th2 type CD+4 T cells,from patients with PV.
2.Expression of Aiolos transcription factor in peripheral blood mononuclear cells (PBMCs) from patients with systemic lupus erythematosus (SLE)
Yangman LIU ; Xiaohong CHEN ; Ying YANG ; Bilan CAO ; Wei YUAN ; Guorong WEN ; Lingfen ZENG
Chinese Journal of Dermatology 2010;43(7):471-473
Objective To investigate the expression of Aiolos transcription factor in peripheral blood mononuclear cells (PBMCs) from patients with systemic lupus erythematosus (SLE),and to explore their clinical significance.Methods Western blot was performed to detect the protein expression of Aiolos transcription factor in PBMCs from 19 patients with active SLE,13 patients with stable SLE and 30 healthy volunteers.The relationship between the expression of Aiolos transcription factor and SLE disease activity index (SLEDAI) was analyzed.Results A significant difference was noted in the relative expression of Aiolos transcription factor between SLE patients and normal controls(0.56±0.17 vs 0.81±0.09,P<0.01) and between patients in active stage and those in stable stage (0.52±0.14 vs 0.65±0.19,P<0.05).In addition,the expression level of Aiolos protein was negatively correlated with SLEDAI in patients (r=-0.65,P<0.01).Conclusion A decrease is observed in the expression of Aiolos transcription factor in SLE patients.which is correlated with the disease activity in SLE.
3.Effects of curcumin on mRNA expression of cytokines related to Tou-like receptor 4 signaling in THP-1 cells
Lixin PENG ; Xinyu LI ; Shasha SONG ; Yongfang WANG ; Lanfang XU
Chinese Journal of Dermatology 2010;43(7):493-496
Objective To investigate the effects of curcumin on mRNA expression of cytokines related to Toll-like receptor (TLR) 4 signaling in THP-1 cells.Methods After pretreatment with different concentrations (50,25,12.5 mg/L) of curcumin or dexamethasone for 12 hours,THP-1 cells were stimulated by lipopolysaccharide (LPS.1 mg/L) for 4 hours followed by the collection of cells.Then total RNA was isolated from these cells and subjected to reverse transcription-polymerase chain reaction (RT-PCR) for the detection of mRNA expression of tumor necrosis factor receptor-associated factor 6 (TRAF6),interleukin-1 receptorassociated kinase (IRAK1) and nuclear factor (NF)-κB.THP-1 cells without pretreatment or stimulation served as negative control,and those only stimulated with LPS served as LPS group.Results After stimulation with LPS (1 mg/L) for 4 hours,the mRNA expressions of TRAF6,IRAK1 and NF-κB were significantly upregulated in THP-1 cells compared with negative control cells (f=38.69,39.13,23.99,all P<0.01).Curcumin of 50 mg/L and 25 mg/L significantly inhibited the mRNA expressions of TRAF6.IRAK 1 and NF-κB upregulated by LPS with an inhibition rate of more than 50% (all P<0.0 1).Conclusions Certain concentrations of curcumin can inhibit the mRNA expressions of TRAF6.IRAK1 and NF-κB.which demonstrates the regulatory effect of curcumin on the mRNA expressions of TLR4 signaling pathway-associated cytokines.
4.Ultrastructure and transcriptional activity of MITF in epidermal melanocytes from patients with vitiligo
Ping WANG ; Weisong HONG ; Li ZHANG ; Jian HONG ; Junhui ZHENG ; Aie XU
Chinese Journal of Dermatology 2010;43(7):485-488
Objective To analyze the relationship of melanocyte ultrastructure and expression of microphthalmia-associated transcription factor (MITF) as well as tyrosinase-related proteins (TRP) transcriptionally modulated by MITF to clinical types and duration of vitiligo.Methods Epidermal sheets were taken by suction blisters respectively from lesional,perilesional,and normal skin of 12 patients with vitiligo vulgaris (VV) and 8 with segmental vitiligo (SV).The duration of vitiligo varied from 3 to 300 months in these patients. Transmission electron microscopy was performed in 10 patients with vitiligo,including 6 cases of VV and 4 cases of SV.Epidermal melanocytes from normal skin of 20 patients were subjected to culture followed by Western blot to detect the expression level of MITF and some molecules transcriptionally modulated by MITF,including tyrosinase (TYR),TYR-related protein-1(TYRP1),and TYR-related protein-2 (TYRP2) in cultured melanocytes.Results Epidermal melanocytes were absent in lesional skin of 7 out of 10 patients observed for ultrastructural alterations,whereas melanocytes with reduced or absent melanin (melanosome) could accidently be seen in lesional skin of 1 patient with short-standing vitiligo and 2 patients with long-standing vitiligo.In perilesional skin.abnormal ultrastructure of melanocytes was found in 3 with a duration of vitiligo less than 15 months among 6 patients with VV,and in 1 out of 4 patients with SV.The down-regulated expression of MITF was consistent with that of TYR,TYRP1 and TYRP2 in cultured epidermal melanocytes from normal skin of patients with VV;in those from patients with SV,the down-regulated expression was observed only in MITF,while the expressions of TYR,TYRP1 and TYRP2 were nearly normal.Conclusion Differences may exist between VV and SV in the ultrastructure as well as mechanisms of transcriptional modulation by MITF in epidermal melanocytes.
5.Proliferation of and production of interferon-γ by drug-specific peripheral T cells from pafients with severe drug eruption
Yang GAO ; Meng FU ; Xiaowei SHI ; Wenjun LIAO ; Lin GAO ; Tianwen GAO ; Gang WANG ; Chengxin LI ; Yufeng LIU
Chinese Journal of Dermatology 2010;43(7):478-481
Objective To detect the proliferation of and production of interferon-γ by drug-specific peripheral T cells from patients with severe drug eruption.Methods Peripheral blood mononuclear cells (PBMCs) were isolated from 10 patients with severe drug eruption,10 patients with mild or moderate drug eruption and 10 normal human controls,stimulated with causative drugs to obtain drug-specific T cells.Then,both PBMCs and drug-specific T cells were stimulated with causative drugs or unrelated drugs followed by the detection of secretion levels of IFN-γ with ex vivo enzyme-linked immunodotting (ELISpot) assay and cultured ELlSpot assav respectively.Results After stimulation with causative drugs,a higher level of IFN-γ was secreted by PBMCs and drug-specific T cells from patients with severe drug eruption compared with those from normal human controls (both P<0.01).and by drug-specific T cells than by PBMCs (P<0.01).The culture with unrelated drugs could neither induce the generation of drug-specific T cells nor promote the secretion of IFN-γ by PBMCs from the patients.Drug-specific T cells still existed in the peripheral blood of 3 patients within 1 to 3 years after recovery of drug eruption.Conclusions There are drug-specific T cells in peripheral blood of patients with severe drug eruption,and they may persist for a certain period of time after recovery of drug eruption.Ex vivo ELISpot combined with cultured ELISpot may be applied to the identification of causative drugs in vivo.
6.Prevalence of vitiligo in China:an epidemiological survey in 6 provinces
Xiaoyan WANG ; Tinglin WANG ; Cheng ZHOU ; Yiwei SHEN ; Xiaolan DING ; Shan TIAN ; Ying LIU ; Guanghui PENG ; June ZHOU ; Shuqi XUE ; Renli WANG ; Ying TANG ; Xuemei MENG ; Guangde PEI ; Yunhua BAI ; Qing LIU ; Hang LI ; Juan DU ; Jianzhong ZHANG
Chinese Journal of Dermatology 2010;43(7):463-466
Objective To investigate the prevalence of vitiligo in China through a multi-center and larse-population epidemiological survey.Methods A community-based survey was conducted in 6 cities from 6 provinces.Cluster sampling method was used.Subjects were required to fulfill the self-report questionnaires and received physical examination by dermatologists.EpiData and SPSS11.5 were utilized for statistical analysis. Results Totally,19 974 patients participated in this study,and 17 345 valid questionnaires were retrieved with a return rate of 86.84%.Of them,122 were found to have vitiligo.The prevalence and standardized prevalence of vitiligo was 0.70% and 0.56% in all patients,0.95% (75) and 0.69% in male patients and 0.50% (47) and O.45% in female patients.respectively.A significant elevation was observed in the prevalence of vitiligo in males than in females (P<0.01).The prevalence of vitiligo was increased with age and peaked in patients aging from 60 to 69 years and those over 70 years.The age at onset of vitiligo varied from 0 to 19 years in 21.85% of these patients,from 20 to 49 years in 47.05%.The most connnon type was focal vitiligo,which accounted for 36.06%,while the rarest type wag segmental type (2.46%).The pesitivity rate of family history of vitiligo was 9.84% in patients and 1.31% in community population (P<0.01).About 31.97% of the patients complained of negative influence of vitiligo on quality of life.Conclusions The standardized prevalence of vitiligo is 0.56%in 6 provinces from China.Males seem to have a higher prevalence of vifiligo than females.
7.Inhibitory effect of live-attenuated Listeria monocytogenes-based vaccines carrying MART-1 gene on mouse malignant melanoma
Ping JIANG ; Yue QIAN ; Aiping FENG ; Siyuan CHEN ; Shujuan CHU ; Li ZHANG ; Yan WU ; Na ZHANG ; Qin LUO
Chinese Journal of Dermatology 2010;43(7):455-459
Objective To investigate the inhibitory effect of live-attenuated Listeria monocytogenes (LM)-based vaccines expressing the gene encoding a melanoma differentiation antigen,MART-1,on malignant melanoma and their mechanism.Methods The constructed plasmid pERL3-MART-1 was used to transform live-attenuated LM by electroporation to construct recombinant LM.i.e.△inlB LM-MART-1 and △actA/△inlB LM-MART-1.The half lethal dose (LD50) of attenuated listeria strains was determined by concentration gradient dilution method.C57BL/6 mice were randomly divided into three groups,namely PBS group,△inlB LM-MART-1 group and △actA/△inlB LM-MART-1 group.Mice were inoculated by intraperitoneal injection of O.1 LD50 of each rLM strain or PBS only.One week later,the mice were injected subcutaneously with 1×105 B16F10 cells(a mouse melanoma cell strain)in 200μl of PBS.Reimmunization was performed on day 14 and 21.Subsequently,the growth of tumor and survival of tumor bearing mice were observed.All mice were killed on day 28,and tumor tissue as well as splenocytes were obtained from these mice for the detection of MART-1 gene expression by real-time quantitative PCR and the percentage of CD4+CD25+T cell by flow cytometry.Results The recombinant △inlB LM-MART-1 and △actA/△inlB LM-MART-1 were constructed successfully.The LD50 of △inlB LM and △actA/△inlB LM was lower than LM-EGDe by 100 and 10 000 times respectively.Compared with PBS,the tumor growth was inhibited with △inlB LM-MART-1 by 46.95%(F=6.3,P<0.05),and by 83.96% with △actA/△inlB LM-MART-1(F=37.8,P<0.01).The relative expression level of MART-1 in △inlB LM-MART-1 group and △actA/△inlB LM-MART-1 group was 8.988±0.207 and 11.315±0.445 times that in PBS group (both P<0.05).The percentage of CD4+CD25+T cells in splenocytes was (2.52±0.20)%,(1.14±0.13)% and (0.44±0.15)% in PBS group,△ialB LM-MART-1 group and △actA/△inlB LM-MART-1 group,respectively;the differences were statistically significant between the three groups (all P
8.Skin cancer and precancerous skin lesions:clinical analysis of 632 cases
Yuanshen HUANG ; Hang LI ; Ping TU ; Xixue CHEN ; Shuxia YANG ; Lingshen WU ; Junyu ZHAO ; Xueying LI
Chinese Journal of Dermatology 2010;43(7):452-454
Objective To investigate the clinical features of skin cancer.Methods Clinical data of skin cancer and precancerous skin lesions confirmed pathologically from 2005 to 2008 in Peking University First Hospital were retrospectively analyzed by using statistical methods.Results A total of 632 cases of skin cancer and precancerous skin lesions were studied.The most common skin cancer was basal cell carcinoma and squamous cell carcinoma (invasive and in situ) which accounted for 29.3%and 24.2%,respectively.The average age at onset was older than 60 years in 55.4%of the patients,between 35 and 59 years in 34.3%,younger than 35 years in 10.3%.The concordance between clinical and pathological diagnosis reached nearly 90.O%for Paget's disease,70.0% for other common skin cancer and precancerous skin lesions.Conclusions Skin cancer and precancerous skin lesions have a predilection for scalp and face.Patients aged from 35 to 59 years account for a significant proportion not only in cutaneous lymphoma but also in melanoma and epithelium-derived nonmelanoma skin cancer.
9.SAPHO syndrome:clinical analysis of 22 cases
Qing YANG ; Lina QU ; Benqing SHI ; Changping YU ; Hongqing TIAN ; Jian PENG ; Fufen ZHANG
Chinese Journal of Dermatology 2010;43(7):449-451
Objective To investigate the clinical features of SAPHO syndrome.Methods Clinical data of 22 cases of SAPHO syndrome were analysed.Results There were 7 males and 15 females among the 22 patients.The average age at onset of cutaneous and osteoarticular lesions was 45 years and 44 years, respectively.Of the 22 patients,21 had palmoplantar pustulosis and 1 had acne fulminans.Anterior chest wall (ACW) was involved in 19 patients,peripheral joints in 4 patients and sacroiliac joints in 2 patients.Osteoarticular manifestations occurred prior to the onset of skin lesions in 10 cases.after that in 9 cases,and simultaneously in 3 cases.The mean interval between the onset of cntaneous and osteoarticular lesions was 2.7 years and the longest interval was 20 years.Conclusions Middle-aged females predominate in patients with SAPHO syndrome seen in dermatological clinics.Palmoplantar pustulosis and ACW involvement are the most common clinical manifestations of SAPHO syndrome.
10.Tumor necrosis factor-α gene-308G/A single nucleotide polymorphism in patients with syphilis
Chun LIU ; Shuzhen QI ; Qianqiu WANG
Chinese Journal of Dermatology 2010;43(7):501-503
Objective To explore the association of tumor necrosis factor(TNF)-α gene-308G/A single nucleotide polymorphism with susceptibility to syphilis.Methods The study includes 56 patients with early symptomatic syphilis,38 with eady latent syphilis,and 102 normal human controls.The-308G/A single nucleotide polymorphism of TNF-α gene was detected in all subjects by real-time fluorescent quantitative PCR.Results The frequency of TNF-α-308 G and A allele was 0.926 and 0.074 in patients with early syphilis,0.941 and 0.059 in the controls,0.929 and 0.071 in patients with early symptomatic syphihs,0.921 and 0.079 in patients with early latent syphilis.No significant difference was found between patients with early syphilis and the controls or between patients with early symptomatic syphilis and those with early latent syphilis in the frequency of either allele (MG) at-308 position of TNF-α gene(all P>0.05).Conclusion There seems to be no evidence for association between TNF-α gene-308G/A single nucleotide polymorphism and susceptibility to syphilis.