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Objective To investigate the effects of different wavelength (595 nm,755 nm and 1064 nm) lasers on the mRNA expression of types Ⅰ and Ⅲ procollagen in cultured fibroblasts.Methods Fibroblasts from Kunming mice were cultured in vitro.They were divided into 595 nm laser group,755 nm laser group,1064 nm laser group and no laser irradiating group.The mRNA expression of the types Ⅰ and Ⅲ procollagen was detected by RT-PCR.Results The mRNA expression level of type Ⅰ procollagen in 1064 nm group was higher than that of 755 nm group,595 nm group and control group (P＜0.05).The expression levels of 755 nm group and 595 nm group were higher than that of control group (P＜0.05).But the difference between 755 nm group and 595 nm group was not statistically significant (P＞0.05).The mRNA expression level of type Ⅲ procollagen in 1064 nm group was higher than that in 755 nm group,595 nm group and control group (P＜0.05).755 nm group had higher expression than 595 nm group and control group (P＜0.05).But the difference between 595 nm group and the control group was not statistically significant (P ＞ 0.05).Conclusions Three wavelength lasers can directly promote mRNA expression of types Ⅰ and Ⅲ procollagen in fibroblasts.595 nm laser mainly promotes mRNA expression of type Ⅰ procollagen,and 755 nm laser promotes more mRNA expression of type Ⅲ procollagen than 595 nm laser.The most mRNA expression of types Ⅰ and Ⅲ procollagen is promoted by 1064 nm laser.
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Objective To investigate the role of epidermal growth factor receptor (EGFR) in the growth and migration of melanoma.Methods A brain-metastatic human melanoma cell line H1,which had been developed in our laboratory,was used in this study.Some H1 melanoma cells were stably transfected with wild-type EGFR via lentiviral vectors to overexpress EGFR (H1EGFRwt),and some H1 cells transfected with green fluorescent protein (GFP) served as the control group (H1GFP).After transfection,the cells were sorted and purified.Then,scratch wound healing assay was performed to estimate the migratory activity of H1 cells; colony-forming assay was conducted to investigate the effects of EGFR on the survival ability and colony-forming ability of H1 cells,and resazurin reduction test was used to evaluate colony formation results; Western blot was carried out to measure the expressions of signaling pathways activated by EGFR.Statistical analysis was done by repeated measures analysis of variance (ANOVA) for the comparison of wound-area healing rate,and by paired t test for the comparison of metabolic activity,between the two groups of H1 cells by using SPSS software version 20.0.Results Flow cytometry showed that H1 cells were successfully transfected with EGFR and GFP respectively,and purified H1EGFRwt and H1GFP cells were obtained after cell sorting.The wound-area healing rate at 6,18,24 and 48 hours after epidermal growth factor (EGF) stimulation was 0.145 ± 0.066,0.479 ± 0.096,0.571 ± 0.198 and 0.672 ± 0.199 respectively for H1EGFRwt cells,0.051 ± 0.036,0.254 ± 0.038,0.303 ± 0.077 and 0.498 ± 0.111 respectively for H1GFP cells.The degree of wound healing in H1EGFRwt cells was significantly higher than that in H1GFP cells (F =68.49,df=5,P ＜ 0.05).Colony-forming assay revealed that the average metabolic activity score of H1EGFRwt cells was significantly higher than that of H1GFP cells (92 225.2 ± 6 632.1 vs.62 935.7 ± 8 159.2,t =2.26,df=9,P ＜ 0.01).Western blot showed that EGF might induce the phosphorylation of downstream signaling proteins such as phosphatidylinositol-specific phospholipase C-γ (PLCγ),signal transducer and activator of transcription 5 (STAT5) and 3 (STAT3) in H1EGFRwt cells,but not in H1GFP cells.In addition,the expressions of phosphorylated serine/threonine protein kinase (pAKT) and mitogen-activated protein kinase (pMAPK) were significantly higher in H1EGFRwt cells than in H1GFP cells after EGF stimulation.Conclusions EGFR may play an important role in the metastasis of H1 melanoma cells,and might serve as a target in the treatment of metastatic melanoma.
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Objective To investigate the expression of mannose-binding lectin (MBL) in lesions of patients with psoriasis vulgaris,and to explore the relationship between MBL and psoriasis pathogenesis.Methods Immunohistochemistry and Western blot were performed to detect the expression of MBL in lesional and normalappearing perilesional skin of 30 patients with progressive psoriasis vulgaris,as well as in normal skin of 30 healthy human controls.Statistical analysis was carried out by t test using SPSS13.0 software.Results Immunohistochemistry showed that MBL was expressed in lesional psoriatic skin,but weakly expressed or absent in normalappearing perilesional skin and normal control skin,with the relative expression level of MBL in lesional skin significantly higher than that in perilesional skin and normal control skin (0.636 7 ± 0.515 1 vs.0.416 3 ± 0.160 1 and 0.381 6 ± 0.310 9,t =2.24,2.32,respectively,both P ＜ 0.05).Western blot revealed a positive expression of MBL protein in all the skin specimens,and the expression intensity of MBL protein in lesional psoriatic skin was significandy increased compared with perilesional psoriatic skin and normal control skin (0.273 1 ± 0.129 4 vs.0.186 3 ± 0.193 1 and 0.149 2 ± 0.268 7,t =2.05,2.28,respectively,both P＜ 0.05).No significant difference was shown in the expression of MBL protein between perilesional psoriatic skin and normal control skin by immunohistochemistry (t =1.51,P ＞ 0.05) or Western blot (t =0.61,P ＞ 0.05).Conclusion There is a high expression of MBL protein in lesions of patients with psoriasis vulgaris,which may be somewhat associated with the pathogenesis of psoriasis.
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Objective To determine the gene polymorphism and serum concentration of mannose-binding lectin (MBL) in patients with psoriasis,and to analyze the relationship between MBL and psoriasis.Methods Totally,67 patients with psoriasis vulgaris and 69 healthy human controls were enrolled in this study.Venous blood samples were obtained from all the subjects.Genomic DNA was extracted,and PCR-restriction fragment length polymorphism (PCR-RELP) analysis was conducted to determine the polymorphism at codon 54 of the MBL gene.Enzyme-linked immunosorbent assay was performed to measure the serum level of MBL.A chi-square goodness-of-fit test was carried out to evaluate Hardy-Weinberg equilibrium,t test to compare the serum concentration of MBL,and chi-square test to compare the frequency of genotypes and alleles of MBL gene codon 54.Results The patients with psoriasis showed higher frequency of GGC/GAC heterozygote but lower frequency of GGC/GGC homozygote (x2 =10.36,P ＜ 0.05),together with increased frequency of GAC allele but decreased frequency of GGC allele (x2 =8.31,P ＜ 0.05),at codon 54 of the MBL gene compared with the healthy controls.The variant allele GAC at codon 54 of the MBL gene was markedly associated with psoriasis (OR =3.383,95％ CI 1.585-7.211,P ＜ 0.05).The serum concentration of MBL was (2.193 7 ± 0.816 3) mg/L in patients with psoriasis,significantly lower than that in the healthy controls ((3.269 5±1.205 8) mg/L,t=6.11,P＜ 0.05).Conclusion MBL might be associated with the pathogenesis of psoriasis to some degree.
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A 16-year-old male presented with a 11-year history of progressively enlarging erythema and crusting on the right cheek.Physical examination revealed an irregularly shaped,sharply marginated,dark erythematous patch sized 6 cm x 10 cm and plaques with mild verrucous proliferation.There were strip-like scar at the margin of lesions and multiple ulcers measuring 0.5 to 1 cm in diameter with firm crusts.No small jellycolored nodules were observed.Direct microscopy of multiple scrapings under the crusts showed many light brown,septate,branching and irregular hyphae.Olivaceous-black woolly colonies grew at 25 C and 35 C on Sabouraud's dextrose agar and potato dextrose agar; flask-shaped conidiogenous cells with funnel-shaped collarettes and ellipsoidal conidia arranged in flower-like shape were observed microscopically.PAS staining showed numerous septate and branching hyphae,pseudohyphae and yeast-like cells.There was a 99.73％ similarity in the species-specific rDNA sequence between the isolate and phialophora verrucosa standard strain CDC-B2152.The patient was diagnosed with cutaneous phaeohyphomycosis caused by Phialophora verrucosa.The lesion subsided after treatment with amphotericin B and itraconazole,but recurred after drug withdrawal.Itraconazole and terbinafine were administered for the retreatment of this patient.
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Objective To study the role of psychological factors in the onset of psoriasis,to investigate the role of monoamine substances in the course of psychological factors-associated psoriasis,and to reveal the psychoneurologic pathogenesis of psoriasis.Methods The psychological factors were assessed with life events scale and questionnaire of A character.The serum levels of norepinephrine(NE),dopamine(DA)and5-hydroxyindoleacetic acid(5-HIAA)were detected with high-performance liquid chromatography(HPLC).Results It was shown that there was an overall tendency of weak A character and moderate depression and anxiety in psoriasis patients.The concentrations of NE,DA and5-HIAA were higher in psoriatic patients than those in normal controls(P