Objective To evaluate the effect of human dermal mesenchymal stem cells (DMSCs) on the expression and secretion of interleukin (IL)-13 by perilesional CD8+ T lymphocytes from patients with vitiligo.Methods Tissue specimens were obtained from the perilesional region of six patients with active vitiligo,and CD8+ T lymphocytes were isolated from both the tissue specimens and peripheral blood of these patients.DMSCs and melanocytes were obtained from the foreskin tissue of healthy males.The 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium,inner salt (MTS) assay was performed to estimate the effect of different concentrations of recombinant IL-13 on the proliferation of melanocytes,reverse transcripition-PCR and Western blotting to detect the mRNA and protein expressions of IL-13 in perilesional and peripheral blood CD8+ T lymphocytes respectively,real-time quantitative reverse transcription (RT)-PCR and enzyme-linked immunosorbent assay (ELISA) to detect the IL-13 mRNA expression in,and IL-13 protein expression in the culture supematant of,CD8+ T lymphocytes before and after coculture with DMSCs,respectively.Statistical analysis was done by t test.Results No obvious changes were observed in the proliferation of melanocytes treated with different concentrations (10,50,100,250,500 μg/L) of recombinant IL-13 for various durations (24,48,72 and 96 hours)compared with untreated melanocytes (all P ＞ 0.05).Both perilesional and peripheral blood CD8+ T lymphocytes expressed IL-13,and the expression was stronger in perilesional than in peripheral blood CD8+ T lymphocytes.A significant decrease was noted in IL-13 mRNA expression (0.100 0 ± 0.002 4 vs.0.383 2 ± 0.018 7,P ＜ 0.05) and protein level in the culture supernatant ((1 509.62 ± 48.44) ng/L vs.(5 507.98 ± 34.11) ng/L,P ＜ 0.05) of CD8+ T lymphocytes cocultured with DMSCs compared with monocultured CD8+ T lymphocytes.Conclusions There is a strong expression of IL-13 by perilesional CD8+ T lymphocytes in patients with vitiligo,which may be inhibited by DMSCs and serve as a target for the treatment of vitiligo.

Objectives To study the difference in the expression of monocyte chemoattractant protein-1 (MCP-1) and soluble intercellular adhesion molecule-1 (sICAM-1) between vitiliginous and non-vitiliginous patches in patients with stable vitiligo, and to compare the number of peripheral regulatory T cells between vitiligo patients in stable stage and normal controls. Methods Suction blister fluid was collected at 1 to 3 h after the suction. The expression of MCP-1 and sICAM-1 in skin tissue fluid was detected with ELISA. The number of regulatory T cells in the blood of vitiligo patients and controls was detected by flow cytometry. Results There was no significant difference in the number of regulatory T cells between vitiligo patients and normal controls. The expression of MCP-1 and sICAM-1 in skin tissue fluid was significantly higher in vitiliginous skin than that in non-vitiliginous patches in patients with common type vitiligo, while there was no significant difference between the two kinds of patches in patients with segmental type vitiligo. Conclusions The immune function is abnormal in vitiliginous skin of the common type vitiligo patients in stable stage, which might explain the lack of success in transplantation for this disease.

Objective To study the effects of Fructus ligustri lucidi and its monomers, tyrosol and oleanotic acid, on the migration of mouse melanoblast cell line (NCCmelb4M5). Methods Cultured NCCmelb4M5 cells were treated with Fructus ligustri lucidi (0.0625, 0.125, 0.25, 0.5, 2 mg/mL), tyrosol (0.02, 0.04, 0.08, 0.16, 0.8 mg/mL) and oleanolic acid (0.0625, 0.125, 0.25, 0.5, 2.5 mg/mL), respectively,for 48 hours followed by the detection of cell proliferation with MTT assay. The working concentration of the three drugs was determined according to the results of MTT assay. Scratch and transwell assays were performed to observe the effect of Fructus ligustri lucidi and its monomers at working concentration on the migration of NCCmelb4M5 cells. Results Based on the results of MTT assay, the working concentration of Fructus ligustri lucidi, tyrosol and oleanolic acid was determined at 0.125 mg/mL, 0.08 mg/mL and 0.0625 mg/mL respectively, and at these concentrations, these drugs exhibited a cytotoxity lower than that of absolute alcohol with no obvious stimulation of cell proliferation. Scratch and transwell assay revealed a promoting effect of both Fructus ligustri lucidi and tyrosol on melanoblast migration (P<0.05), while oleanolic acid had little effect on melanoblast migration. Conclusions The extract of Fructus ligustri lucidi has a significant stimulatory effect on the migration of mouse melanoblasts, and tyrosol may be an active component of Fructus ligustri lucidi associated with confirmative effect on migration of mouse melanoblasts.

Objective To evaluate the effect of injuries on monobenzone-induced vitiligo-like depigmentation in mice.Methods Forty C57BL/6 mice were randomly and equally divided into four groups:negative control group topically treated with vaseline cream,model group induced by topical monobenzone (40％) cream,acupuncture group receiving acupuncture treatment (15 times) once every three days,and acupuncture combined with monobenzone group receiving both monobenzone induction and acupuncture treatment.The treatment lasted 50 days and mice were sacrificed 15 days after the end of treatment.Hair decolorization was observed with naked eyes,and skin decolorization with reflectance confocal microscopy (RCM) on a daily basis.Tissue specimens were obtained from depigmented skin at monobenzone-uninduced sites,and subjected to hematoxylin and eosin (HE) staining for the cvaluation of lymphocytic infiltration as well as immunofluorescence staining for the detection of CD8+ T cell expression.Statistical analysis was done by t test.Results Varying degrees of depigmentation were observed in both monobenzone-induced and-uninduced sites in both the model group and acupuncture combined with monobenzone group,and the latter group showed earlier,larger and more stable depigmentation than the former group.At 15 days after the end of treatment,the decolorization area index in the model group and acupuncture combined with monobenzone group was 3.45 ± 0.17 and 3.90 ± 0.25 at monobenzone-induced sites respectively(t =7.433,P ＜ 0.05),1.90-± 0.12 and 2.85 ± 0.27 at monobenzone-uninduced sites respectively (t =7.529,P ＜ 0.05).Significant differences were observed in the fluorescence intensity of CD8 + T cells at monobenzone-uninduced depigmented sites between the model group and acupuncture combined with monobenzone group (175.528 ± 10.711 vs.645.928 ± 12.652,t =8.105,P ＜ 0.05),and there was a more evident infiltrate with lymphocytes and CD8+T cells in the monobenzone-uninduced depigmented sites in the acupuncture combined with monobenzone group.Conclusion Local destruction of skin barrier may promote monobenzone-induced vitiligo-like decolorization in mice.

Objective To evaluate the performance of the hypersensitive C-reactive protein(hs-CRP) rapid quantitative chemilu-minescent detection kit .Methods According to National Committee for Clinical Laboratory Standards (NCCLS) EP10-A2 docu-ment ,hs-CRP rapid quantitative chemiluminescent detection kit was employed to measure the CRP at low ,medium and high con-centration levels of quality control serum .Bias ,total imprecision and their slope rates ,intercepts ,carryover ,non-linearity and drift were calculated ,and its clinical acceptability was evaluated .Results Bias and total imprecision of hs-CRP rapid quantitative chemi-luminescent detection kit were within the allowable ranges ,the average values of slope rates ,intercepts ,carryover ,non-linearity and drift were 1 .005 7 ,0 .537 8 ,0 .789 6% ,0 .019 2 ,0 .036 0 ,respectively ,the differences showed no statistically significance ( P>0 .05) .Conclusion hs-CRP rapid quantitative chemiluminescent detection kit has good accuracy and precision ,stable perform-ance ,and consistent with the clinical testing requirements .

ObjectiveTo assess the relationship of interleukin-17(IL-17) and transforming growth factor-β(TGF-β) with the development of vitiligo.MethodsEnzyme-linked immunosorbent assay (ELISA) was carried out to measure the levels of IL-17 and TGF-β in sera from 120 patients with vitiligo and 60 healthy controls.The correlations of serum IL-17 and TGF-β levels with patients' gender,stage and duration of disease,involved body area and presence of family history were assessed.ResultsThe level of serum IL-17 was significantly higher in patients with active vitiligo than in the controls and patients with stable vitiligo (both P ＜0.05).With the rise in involved body area,the level of serum IL-17 gradually increased (x2 =12.656,P ＜0.05).The level of TGF-β in patients with active vitiligo was a little higher than that in the controls and patients with stable vitiligo,with no significant difference between these groups(both P ＞ 0.05).Conclusions The levels of serum IL-17 and TGF-β are somewhat correlated with the activity of vitiligo,and both of them may play a certain role in the pathogenesis of vitiligo.

Objective To evaluate the protective effects of tea polyphenols against the destruction of melanocytes by CD8+ T cells from vitiligo patients.Methods Skin tissue was resected from the margin of vitiligo lesions followed by the isolation and culture of CD8+ T lymphocytes,and from the normal skin of vitiligo patients followed by the isolation and culture of melanocytes.Flow cytometry was carried out to evaluate the purity of CD8+ T cells.The melanocytes were cocultured with the CD8 + T cells at different ratios followed by the evaluation of killing effect of CD8+ T cells.Various concentrations (200 and 400 μg/ml) of tea polyphenols were added into the co-culture system of CD8+ T cells and melanocytes at a ratio of 5 ∶ 1 followed by an additional culture of 48 hours.Then,flow cytometry was performed to detect the apoptosis in melanocytes in the coculture system.Results CD8+ T lymphocytes were successfully obtained from the marginal area of vitiligo lesions with a purity of more than 90％,which highly expressed the antigens CD137 and CD69.The coculture with CD8+ T cells markedly accelerated the apoptosis in melanocytes,while the accelerative effect was inhibited by tea polyphenols of 200 and 400 μg/ml.Conclusions The CD8+ T cells infiltrating the edge of vitiligo lesions display a potential destructive effect on autologous melanocytes from vitiligo patients,and tea polyphenols have a protective effect against the destruction of melanocytes by CD8+ T cells.

Objective To detect the levels of soluble vascular cell adhesion molecule-1 (sVCAM-1) and soluble intercellular adhesion molecule-l (sICAM-1) in the sera of patients with systemic lupus erythe-matosus (SLE) and their clinical significance was analysed. Methods Serum level of sVCAM-1 and sICAM-1 of 30 controls and 60 SLE patients were measured by enzyme linked immunosorbent assay (ELISA). Results 1 Serum levels of sVCAM-1 were significantly increasd in patients with SLE compared with those in normal controls (P

Objective To investigate the relationship between the serum level of soluble vascular cell adhesion molecule-1(sVCAM-1) and soluble intercellular adhesion molecule-1(sICAM-1) and the disease activity in systemic lupus erythematosus(SLE). Methods The serum concentrations of sVCAM-1 and sICAM-1 were measured by ELISA in 60 SLE patients and age- and sex-matched normal controls. Results ① Serum levels of sVCAM-1 and sICAM-1 were significantly increased in SLE patients compared with those in normal controls (P

Objective To investigate the relationship between the activity of vitiligo and the serum levels of IgG and IgM anti-tyrosinase antibodies and sICAM-1. Methods Anli-tyrosinase, anti-melanocyte antibodies and sICAM-1 were detected by enzyme-linked immunosorbent assay. Results The average titres of IgG and IgM antibodies in the sera of patients with vitiligo were 0.316 and 0.238 respectively, which were significantly higher than those of the healthy control (both P 0.05) in the serum level of sICAM-1 between the healthy control and patients with stable vitiligo or localized vitiligo. Conclusions Both the serum levels of IgG and IgM anti-tyrosinase antibodies and sICAM-1 have a stong correlation with the activity and severity of vitiligo.