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Objective Through clinical observation and statistics, to get the best curative effect of surgical operation in treating pigment nevus method and provide clinical guidance. Methods We reviewed of face and neck patients (1100 patients) with pigmented nevus in the department in the department of the dermatology,plastic surgery from January 2013 to October 2015, two different methods was designed on each parts and effect of the treatment, especially satisfaction degree was analyzed by statistical methods.Results In 100 cases of patients, only 20 patients had mild scar hyperplasia at the neck incision and the rest of the patients were satisfactory. For special parts such as mouth,nose and eye around,along thedirection of the muscle, arc and along the direction of dermatoglyph incision was designed respectively, patients obtained with higher postoperative satisfaction (P<0.05) . Conclusion In pigmented nevus of face and neck surgery treatment, surgical incision design requires dynamic and static combining method,incision design is important for the postoperative effect and patients' satisfaction.
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Objective To estimate the effect of different doses of ultraviolet (UV) radiation on the proliferation of and apoptosis in kertatinocytes,as well as on the expression of p53,matrix metalloproteinase-2 (MMP2) and-9 (MMP9) in actinic keratosis (AK) lesions and normal human skin.Methods Tissue specimens were obtained from the lesions of 20 patients with AK and sun-exposed normal skin of 20 healthy human subjects,and subjected to an air-exposed culture.Each of the specimens was divided into 4 areas to remain untreated (control area) or be irradiated with UV of 5,10 and 20 J/cm2 (irradiated areas) for 4 consecutive days.After another 24-hour culture,the tissue cultures were collected followed by the evaluation of apoptosis in and proliferation of keratinocytes by using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) and Ki-67 staining,and determination of mRNA and protein expressions of p53,MMP2 and MMP9 by using real time PCR and immunohistochemistry respectively.Results A statistical increase was observed in the percentage of apoptotic cells in the normal skin irradiated with UV of 10 and 20 J/cm2 (46.8％ ± 2.1％ and 56.7％± 2.4％,both P ＜ 0.05) and in the AK lesions irradiated with UV of 20 J/cm2 (43.5％ ± 1.5％,P ＜ 0.05)compared with the corresponding unirradiated tissues.The normal skin showed a higher percentage of apoptotic cells than the lesional skin after irradiation with UV of 10 and 20 J/cm2 (both P ＜ 0.05).The percentage of Ki67-positive cells was significantly decreased in the normal skin after irradiation with UV of 20 J/cm2 (3.34％ ±0.76％,P ＜ 0.05),but experienced no statistical changes in the lesional skin after different doses of UV irradiation (all P ＞ 0.05).There was a statistical elevation in the expression of p53 mRNA (5 J/cm2:1.106 ± 0.025,10 J/cm2: 1.259 ± 0.045,20 J/cm2:1.425 ± 0.053,all P ＜ 0.05) and protein(10 J/cm2:0.1169 ± 0.0032,20 J/cm2:0.1454 ± 0.0047,both P＜ 0.05) in the normal skin,but a statistical reduction in the expression of p53 mRNA(10 J/cm2.0.611 ± 0.050,20 J/cm2:0.578 ± 0.070,both P ＜ 0.05) and protein (20 J/cm2:0.0404 ± 0.0027,P＜ 0.05) in the lesional skin after irradiation compared with the corresponding unirradiated skin tissues.Further more,a statistical increment was observed in MMP2 mRNA and protein expression in normal skin irradiated with UV of 10 J/cm2 (1.086 ± 0.013,0.0843 ± 0.0024,respectively,both P ＜ 0.05) and 20 J/cm2 (1.417 ± 0.036,0.1236 ±0.0042,respectively,both P ＜ 0.05) and in lesional skin irradiated with UV of 20 J/cm2 (1.296 ± 0.028,0.0744± 0.0032,respectively,both P ＜ 0.05),as well as in MMP9 mRNA and protein expression in normal skin irradiated with UV of 20 J/cm2 (1.395 ± 0.026,0.3065 ± 0.0162,respectively,both P ＜ 0.05) and in lesional skin irradiated with UV of 10 J/cm2 (1.298 ± 0.035,0.0992 ± 0.0053,respectively,both P ＜ 0.05) and 20 J/cm2(1.286 ± 0.032,0.1010 ± 0.0063,respectively,both P ＜ 0.05) compared with the corresponding unirradiated tissues.Conclusion Ultraviolet may accelerate the progression of AK by down-regulating p53 expression but up-regulating MMP2 and MMP9 expression.
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Objective To detect the expression and content of decorin in fibroblasts of keloid to deeply reveal the mechenism and the role of decorin plays in scar formation.Methods Fibroblasts of keloid,normal scar and normal skin were cultured in vitro,and the morphology,activity,apoptosis of fibroblast were observed under light microscope and electron microscope; the mRNAs of decorin and TGF-β1 were detected and analyzed with real-time fluorescent quantitative-PCR (FQ-PCR).Results Fibroblasts of keloid showed irregular morphology,larger size and disorder arrangement.There were a large number of mitochondria,swelling rough endoplasmic reticulum,and euchromatin-rich in nucleus of fibroblasts,suggesting the protein synthesis of keloid fibroblast was very active.Compared with normal skin,the expression of decorin was significantly lower in keloid fibroblast; On the contrary,the expression of TGF-β1 was significantly higher in keloid fibroblast than in normal scar and normal skin.Conclusions Compared with normal skin,the expression of decorin in keloid fibroblast is significantly lower.Lower content of decorin in early stage of wound healing may induce weakly suppression of proliferation and synthesis of fibroblast,and up-regulate the activity of TGF-β1,which promotes the proliferation,migration and excessive collagen synthesis of the fibroblast of keloid.Thus,decorinis an suppressor factor of keloid formation.
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Objective To evaluate the performance of actinic keratosis (AK) tissue as a culture model for the study of interference in transduction pathway,and to explore the mechanism underlying the p53 regulation though TGFβ1/Smads pathway by using the tissue culture model.Methods Twenty-five skin samples from the lesions of patients with AK were cultured,and divided into 5 groups to be treated with TGFβ1 of 10 μg/L for 24 and 48 hours,the tran sforming growth factor (TGF) β1 receptor kinase inhibitor SB431542 of 10 μmol/L for 24 and 48 hours,respectively,or remain untreated.Real time PCR and Western blot were performed to quantify the mRNA expression of p53 and protein expression of p53 and phosphorylated Smad2 in these tissue specimens respectively.Results A significant elevation was observed in the expressions of p53 mRNA ( 13.4968 ± 0.9903 vs.1,P ＜ 0.05) and phosphorylated Smad2 (0.700 ± 0.023 vs.1,P ＜ 0.05) in AK tissues after treatment with TGFβ1 for 24 hours,and in the expressions of p53 mRNA (13.3882 ± 1.6772 vs.1,P ＜ 0.05) and protein (1.009 ± 0.001 vs.0.512 ± 0.005,P ＜ 0.05) after treatment with TGFβ1 for 48 hours,compared with the untreated AK tissues.No significant differences were observed in the expression of p53 protein between the AK tissues treated with TGFβ1 for 24 hours and 48 hours (P ＞ 0.05).SB431542 induced a statistical reduction in the level of phosphorylated Smad2 at 48 hours (0.116 ± 0.003 vs.0.306 ± 0.023,P ＜ 0.05),but no significant changes were observed in the expression of p53 mRNA or protein after SB431542 treatment for 24 or 48 hours.Conclusions AK tissue cultures can serve as a model for the study of interference in signal transduction pathway.TGFβ1 might regulate the expression of p53 protien through Smads pathway in AK.
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BACKGROUND: Nerve growth factor is secreted and synthetized by a variety of cells, such as inflammatory calls and repairing calls, its biological effects are diverse and closely related to the process of wound repair, but its mechanism is not yet clear.OBJECTIVE: To observe the influence of nerve growth factor on the biological characteristics of scar fibroblasts.METHODS: Eight clinical surgical resection specimens, including 5 face and neck hyperplastic scar or keloid specimens, did not receive any treatment; three were prepuce specimens following circumcision (normal tissue). By use of tissue block method, the scar and normal skin fibroblasts were cultured, followed by digestion passage. The scar tissue and normal tissue flbroblasts at 3-6passages in the logarithmic phase were seeded in 96-well plate and divided into the experimental group (scar flbroblest group) and the control group (normal skin fibroblasts group), with two parallel holes in each group were added with 3,33, 0.33 mg/L nerve growth factor, 50 μL. Inverted microscope was used to observe fibroblast morphology. At 24, 48, 72 hours after culture, the absorbanca value was measured using MTT. Fibroblast DNA content and cell apoptosis were determined by flow cytometry.RESULTS AND CONCLUSION: The fibroblasts were adherent cells, the scar and normal skin tissues were shown to cell free out of tissue block and gradual expansion at 4-6 days after incubation. Compared with normal skin fibroblasts, the pathological scar fibroblasts became larger, irregular shape and arrangement. MTT results showed that nerve growth factor could promote the normal and hypertrophic scar fibroblasts growth, which becomes more apparent. Flow cytometry results showed that by adding nerve growth factor, the percentage of scar fibroblasts at proliferating S-G_2-M phase was higher than that in the control;group; with a Iower level of apoptosis. It is indicated that nerve growth factor plays an obviously promoting role on normal and scar skin fibroblasts growth and proliferation, especially on the scar skin.
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Objective To explore the mechanism of cytokines for the scars,and to study the effect of platelet derived growth factor(PDGF)on the biological behavior of fibroblasts in scars.Methods Fibroblasts of scars and normal skins were cultured in vitro.The results were observed and analyzed by light inverted microscopy(LM),and 3-(4,5-dimethyl-thiazol-2-yl)-2,5 ciphenyl tetrazolium bromide (MTT)assay.The effects of PDGF on the biological behaviors of fibroblasts of scars were also determined. Results In vitro study,using LM,FCM and MTT assay,showed that proliferation of fibroblasts were inereased significantly when PDGF was added to the cultures,as compared to the control groups.Conclusions PDGF can increase fibroblast proliferation.These results demonstrate that PDGF is beneficial for wound healing at early stage.