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Objective To measure the expressions of miR-203 and its downstream target genes p63 and survivin in psoriasis vulgaris lesions.Methods Tissue specimens were collected from lesions of 30 patients with psoriasis vulgaris and normal skin of 30 healthy human controls.Real-time PCR was performed to detect miR-203 mRNA expression with U6 as the internal control,as well as p63 and survivin mRNA expressions with glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as the internal control,and Western blot to measure the protein expressions of miR-203 targets p63 and survivin.Statistical analysis was carried out by t test for intergroup comparisons and by Pearson correlation analysis for the analysis of correlation between miR-203,p63 and survivin expressions in psoriasis vulgaris lesions.Results Compared with the normal control skin,psoriasis vulgaris lesions showed significantly decreased mRNA expression level (2-△△C△△Ct) of miR-203 (0.41 ± 0.11,t =3.16,P ＜ 0.05),but increased mRNA expression levels of p63 (4.79 ± 0.63,t =4.72,P＜ 0.05) and survivin (3.43 ± 0.46,t =4.35,P＜ 0.05).The protein expression levels of p63 and survivin in these lesions were (2.40 ± 0.23) times (t =3.87,P ＜ 0.05) and (3.49 ± 0.14) times (t =4.36,P ＜ 0.05) those in the normal control skin respectively.In psoriasis vulgaris lesions,the mRNA expression level of miR-203 showed a significantly negative correlation with that of survivin (r =-0.36,P ＜ 0.05) and p63 (r =-0.43,P ＜ 0.05).Conclusion miR-203 and its downstream target genes p63 and survivin may participate in the occurrence of psoriasis vulgaris.
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Objective To measure the expression of activin receptor-like kinases 1(ALK1)in dermal fibroblasts from patients with systemic scleroderma(SSc)and to estimate its role in the production of fibronectin and plasminogen activator inhibitor-1(PAI-1).Methods Dermal fibroblasts were isolated from the lesions of 12 patients with SSc as well as the normal skin of 14 healthy controls,and subjected to a primary culture.The third-passage fibroblasts were used in the next experiment.Western blot and indirect immunofluorescence technique were utilized to quantify the expression of ALK1.A specific siRNA targeting ALK1 was designed,constructed,and transiently transfected into the control dermal fibroblasts,which were then classified into 2 groups to be cultured with or without the presence of transforming growth factor(TGF)-β1 for 72 hours followed by the detection of fibronectin and PAI-1 expression with Western blot.Results As Western blot and direct immunofluorescence technique showed,both control and SSc fibroblasts showed an expression of ALK1 in the cytoplasm and membrane,and the expression intensity of ALK1 in SSc fibroblasts was significantly higher than that in the control fibroblasts(1.97 ± 0.05 vs.1.12 ± 0.03,t =50.96,P ＜ 0.05).The expression of ALK1,fibronectin and PAI-1 was decreased by 90％,58％ and 31％ respectively in specific siRNA-transfected SSc fibroblasts compared with the control siRNA-transfected fibroblasts.TGFβ1 significantly increased the expression of ALK1,fibronectin and PAI-1 in the control siRNA-transfected fibroblasts,but the increase was markedly inhibited by the siRNA-targeting ALK1.Conlusion TGFβ1 can promote the production of fibronectin and PAI-1 via ALK1 in fibroblasts,and ALK1 may be involved in the development of sclerosis in SSc.
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Objective To investigate the possibility of hair follicle reformation induced by hair follicle bulb cells implanted into collagen/chitosan porous scaffolds in vivo, and to observe the angiogenesis in implanted scaffolds.Methods Hair follicle bulb cells obtained by enzyme digestion from the hack skin of C57BL/6J mice were implanted into collagen/chitosan porous scaffolds followed by 2-week organotypie culture.Then,these collagen/chitosan porous scaffolds were transplanted subcutaneously into the dorsal skin of nude mice.Those nude mice transplanted with empty collagen/chitosan porous scaffolds served as the controls.The growth of hair was observed with naked eyes.Six weeks after the transplantation,skin samples were obtained from the recipient site and subjected to histological examination.Results Five weeks after the transplantation,hair growth was observed in the dorsal skin of nude mice.Six weeks later,histological examination revealed fully differentiated hair follicles and vessel-like structures in the center of collagen/chitosan porous scaffolds.However,the transplantation with empty collagen/chitosan porous scaffolds failed to elicit the same response.No hair or follicles were observed in the control mice alpng with small number of vessel-1ike structures.Con-clusions Hair follicle bulb ceils implanted into collagen/chitosan porous scaffolds in vivo could induce hair follicle reformation and promote the formation of vessel-like structure in the scafffold center.
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Objective To investigate the performance of surgical management in facial skin malignancies.Methods From January 2000 to December 2006,65 patients with facial skin malignancies,including47 cases of basal cell carcinoma.10 cases of squamous cell carcinoma,3 cases of dermatofibrosarocoma protuberans,2 cases of malignant melanomas,and one case of malignant acanthoma,hemangioendotheliosar-coma and sebaceous carcinoma,respectively,were collected and managed with wide resection followed by reconstruction.In order to achieve a thorough resection,frozen sections were prepared and subjected to pathological examination during the operation process to ensure the margins of resection were free of malignancy.Reconstruction was carried out by direct closure,or with local random flaps,extended flaps,free skin grafts.Resuits All defects were managed by one-stage reconstruction.The survival rate of skin flaps/grafts was 100%,and a satisfactory appearance and function was achieved.During the follow-up from 6 months to 5 years,local relapse was observed in one patient with basal cell carcinoma and one with squamous eell carcinoma,lymphatic metastasis in one with squamous cell carcinoma.Distant metastasis occurred in a patient with malignant melanoma.who died consequently.Conclusions Thorough resection is the key to prevent relapse of facial skin malignancies after surgery.Appropriate reconstruction may favor the restoration of facial appearance,and local random flaps appear to be the best reconstruction strategy.
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Objective To address the question whether circulating T cells from the patients with primary and metastatic malignant melanoma show altered chemotaxis to monocyte chemotactic protein 1 (MCP 1) and its relation to tumor infiltrating lymphocyte (TIL) and metastasis. Methods Chemotactic responsiveness of T cells towards MCP 1 and immuno histochemistry study were investigated in patients with primary and metastatic melanoma compared to patients with basal cell carcinoma and healthy persons. Results T cells from primary and metastatic melanoma patients showed a significantly decreased chemotactic migration towards MCP 1 and that T cells from patients with basal cell carcinoma showed normal chemotactic response. Immuno histochemistry study showed that there was no correlation between the density of TIL and the decreased chemotaxis of circulating T cells to MCP 1 in patients with primary melanoma. Conclusion Circulating T cells from patients with primary and metastatic malignant melanoma show a MCP 1 specific decrease in chemotactic migration, this may be due to abnormal expression or modulation of MCP 1 receptor on these cells.
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Objective To investigate the correlation between expression of four adhesion molecules in sera and in skin lesions of patients with psoriasis vulgaris, as well as their relevance to clinical severity of the disease. Methods Enzyme-linked immunosorbent assay (ELISA) was used to determine the serum levels of ICAM-1, ICAM-3, VCAM-1 and ELAM-1 in 36 patients with plaque type psoriasis vulgaris and 36 healthy controls. Avidin biotin immunoperoxidase staining system was employed to quantitate expression of the four adhesion molecules in psoriatic skin lesions before and after treatment. Results Significantly increased expression of four adhesion molecules was found in psoriatic skin lesions (P
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Objective To study the relationship between the in situ expression of infiltrating T lymphocyte subsets and leukocyte adhesion molecules in psoriatic patients. Methods Immunohistochemical technique was employed to study the expression of infiltrating T lymphocyte subsets (CD3, CD4, CD8) and leukocyte adhesion molecules (ICAM 1, ELAM 1, VCAM 1) in the lesional and non lesional psoriatic skin. Results There was a significant increase of T lymphocyte subsets (CD3, CD4 and CD8) and adhesion molecules (ICAM 1, ELAM 1, VCAM 1) in the lesional skin than those in non lesional skin in psoriasis, and that there is a significant positive correlation between the density of infiltrating T lymphocyte subsets and the intensity of in situ expression of adhesion molecules. In comparison to the materials from normal controls, the upregulation expression of adhesion molecules was observed in non lesional skin and normal appearance skin after short duration topical treatment with corticosteroid. Conclusion Increased upregulation expression of adhesion molecules is closely associated with the infiltration of T lymphocytes in lesional skin of psoriasis and may be one of the factors for the recurrence of psoriasis.
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Objective To study the chemotactic activity of T lymphocytes from patients with psoriasis to proinflammatory cytokines in vitro and its role in the pathogenesis in psoriasis. Methods A 48 microchemotaxis chamber was employed to determine T cell chemotactic activity. In addition, the expression of T cell activation markers such as HLA DR and interleukin 2 receptor was analysed with fluorescence activated cell sorting technique and serum IL 8 level was measured with ELISA. 45 patients with psoriasis (23 patients with severe psoriasis and 22 patients with mild psoriasis) and 21 patients with atopic dermatitis were investigated, and 20 healthy controls were tested equally. Results ①T cell chemotactic responses were significantly decreased in patients with severe psoriasis and atopic dermatitis as compared to healthy controls. ②Increased expression of activation markers such as HLA DR and interleukin 2 receptor was demonstrated in circulating T cells from severe psoriatic patients and atopic dermatitis patients in comparison to healthy controls. ③Serum IL 8 level was significantly increased in patients with psoriasis and atopic dermatitis. Conclusion The in vitro chemotactic response of circulating T cells in patients with severe psoriasis to IL 8 is significantly impaired, furthermore, the in vivo activation state of T lymphocytes in these patients and increased level of serum IL 8 seem to be associated to the decreased in vitro T cell chemotactic response.
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Objective To observe the ability of cultured dermal papilla cells(DPCs) to induce hair follicle regeneration and to sustain hair growth in vivo and in vitro. Methods The expression of basic fibroblast growth factor (bFGF), endothelin-1 (ET-1) and stem cell factor (SCF) in different passages of DPCs, and their possible effects on biologic behaviour of DPCs were measured by in situ hybridization and immunohistochemistry. Hair follicle regeneration induced by DPCs in hair follicle organotypic culture model and the model implantated into nude mice were studied. Results The expression of ET-1 and SCF in the early passages of cultured DPCs was strong, but became weak and even negative after 6 passages. Hair follicle-like structures were formed in the hair follicle organotypic cultures, composed of DPCs and hair follicle epithelium cells. When the hair follicle organotypic cultures were implanted into the subcutis of nude mice, relatively intact hair follicles were formed. Injection of the early passage DPCs mixed with hair follicle epithelial cells into the subcutis of nude mice resulted in the formation of hair follicle-like structures, while the structures were not formed after the injection of the mixture of hair follicle epithelial cells with dermal sheath fibroblasts or with scalp fibroblasts. There was a positive correlation between the expression levels of ET-1 and SCF in DPCs and the ability of DPCs to induce hair follicle regeneration . Conclusions Cultured DPCs can induce hair follicle regeneration and sustain hair growth in vivo and in vitro. Moreover, the expression levels of ET-1 and SCF are positively correlated with the ability of DPCs to induce hair follicle regeneration.