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.

Objective To evaluate the clinical value of laparotomy,transvaginal surgery,and lapaioscopic surgery in treating benign ovarian cyst.Methods We retrospectively analyzed 227 patients with benign ovarian cyst who underwent laparotomy,transvaginal surgery,or laparoscopic surgery.The duration of the operation,amount of blood loss,hospitalization time,anal exhaust time,temperature recovery time.outof-bed time,usage of analgesics,postoperative morbidity,and hospitalization cost were analyzed.Results There were significant differences in above indicators between the patients undergoing transvaginal surgery or laparoscopic surgery and those undergoing laparotomy.Conclusion Transvaginal surgery and laparoscopic surgery are minimally invasive and safe in treating benign ovarian cyst.These 3 surgical approaches have distinct advantages and disadvantages.The clinicians should choose the appropriate surgical approach for benign ovarian cyst according to the patient's condition.

Objective To evaluate the efficacy and safety of conventional 2940 nm Er∶YAG laser and pixel laser combination therapy.Methods From Jan.2010 to Jun.2013,32 cases of different types of superficial scars had been treated with this combination therapy.All patients were treated with the low-power 2940 nm Er∶YAG laser,and then immediately with pixel laser at higher power.The morphological changes of the scar before and after treatment,and adverse reaction were recorded.The satisfaction of patients was collected.Results Based on imaging data,significant efficacy was achieved in 14 cases (43.8％),effective in 12 cases (37.5％),relatively effective in 5 cases (15.6％) and ineffective in 1 case (3.1 ％).The total efficacy rate of combined therapy reached 95.3 ％.Very satisfaction to the treatment was 13 cases (40.6％),satisfaction in 10 cases (31.2％),relative satisfaction in 7 cases (21.9％) and no satisfaction in 2 cases (6.3％).The total satisfactory rate reached 93.8％.The only observed side effect was the erythema reaction at irradiated area.Conclusions The combined therapy of conventional 2940 nm Er∶YAG laser and pixel laser is an uncomplicated and safe way to treat superficial scars with little reverse reaction.

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.

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

[ Objective ] To screen the duck model with congenital infection of duck hepatitis B virus (DHBV) by polymerase chain reaction (PCR) method. [Methods] Serum DHBV-DNA level in one-day-old ducklings was detected by PCR method and was compared with that by Dot-blot method. Ducklings with serum DHBV-DNA being negative confirmed by PCR method were inoculated DHBV-DNA positive serum to establish acquired infection models. Pathological features of liver tissues in the congenital infec tion model and the acquired infection model were also observed. [Results] Sensitivity and specificity of PCR detecting serum DHBV-DNA were superior to those of Dot-blot method. In the congenital infection model, viremia maintained long time, the titer of serum DHBV-DNA was high and the inflammatory af fection in liver tissues was slight as compared with those in the acquired infection model. [Conclusion] The duck model with congenital infection of DHBV screened by PCR method is more suitable for the phar macological and pharmacodynamic research of drugs for chronic hepatitis B.

Objective To explore the influence of interbody fusion intervertebral fixation on adjacent joint degeneration in patients with lumbar diseases .Methods 56 patients with lumbar diseases who were taken interbody fusion intervertebral fixation were selected as the research subjects ,and the adjacent intervertebral joint degeneration in patients were followed up .Results 56 patients were followed up for 24-72 months,the average time was (41.2 ± 8.3)months.The new clinical symptoms were occurred in 12 cases(21.4%),including 8 cases of low back pain, 3 patients with leg pain,1 case of lumbocrural pain.Radiographic changes occurred in 9 cases(16.1%),9 cases of patients with new clinical symptoms ,including 5 cases of osteophyte formation or exacerbation ,2 cases for adjacent segment instability ( I degree of vertebral spondylolisthesis ) ,1 case of small joint hyperplasia stage spinal stenosis , 1 case of protrusion of intervertebral disc .In 9 patients with imaging performance ,internal fixation of upper adjacent segment degeneration in 7 cases,internal fixation of the adjacent segment degeneration in 2 cases.Conclusion Inter-body fusion intervertebral fixation in the treatment of lumbar diseases ,adjacent joint is prone to degeneration ,so the destruction of adjacent joint operation should be reduced .

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.

BACKGROUND:Both high glucose and lipopolysaccharide have been proved to promote the apoptosis of human periodontal ligament fibroblasts (HPLFs), but their interactions on the HPLF apoptosis in vitro have not yet been reported. OBJECTIVE:To investigate the effect of different concentrations of lipopolysaccharide and high glucose on the proliferation, apoptosis and the expression levels of Bax and Bcl-2 in HPLFs in vitro. METHODS:The primarily cultured HPLFs were identified. The 5-8 generations of HPLFs were col ected and used in the subsequent experiment. The HPLFs were cultured in different concentrations of glucose (5.5 and 25 mmol/L) and lipopolysaccharide (0, 1 and 10 mg/L) for 24 and 48 hours, respectively. RESULTS AND CONCLUSION:Lipopolysaccharide (10 mg/L) could significantly inhibit the cel proliferation, promote the cel apoptosis, upregulate the expression levels of Bax and Bcl-2 mRNA and induce a significant decrease in Bcl-2/Bax ratio in the cel s cultured with 5.5 mmol/L glucose (P<0.05). The lipopolysaccharide-induced suppression of cel proliferation, cel apoptosis, the expressions of Bax and Bcl-2 mRNA as wel as decrease in Bcl-2/Bax ratio were significantly strengthened in the HPLFs treated with 25 mmol/L glucose (P<0.05). Analysis of variance found that high glucose and lipopolysaccharide had a significant interaction on the cel apoptosis (P<0.05). These results reveal that lipopolysaccharide-induced suppression of cel proliferation, cel apoptosis and the expressions of Bax and Bcl-2 mRNA are augmented in HPLFs cultured under high glucose condition, indicating lipopolysaccharide and high glucose interactively act in inducing cel apoptosis.

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.