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A 22-year-old female patient presented with skin flushing in the bilateral legs for 4 years, which gradually spread throughout the whole lower limbs and forearms 6 months ago. Skin examination showed diffuse flushing and dilated capillaries in the lower limbs and both forearms, and the flushing faded after a press. Histopathological examination of the skin lesion on the leg showed hyperkeratosis in a basket-like shape, increased pigmentation in the basal layer, infiltration of the superficial dermis with scattered lymphocytes, with no obvious red blood cell overflow; periodic acid-Schiff staining showed thickened and homogeneous deposits around the blood vessels; immunohistochemical staining showed thickened blood vessel walls and positive staining for type Ⅳ collagen. Diagnosis: cutaneous collagenous vasculopathy.
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Objective To evaluate the effect of selenomethionine (Se-Met) against ultraviolet B (UVB)-induced oxidative damage to human HaCaT keratinocytes,and to explore its possible mechanisms.Methods Cultured HaCaT cells were divided into several groups:normal control group receiving no treatment,Se-Met groups treated with Se-Met at concentrations of 1,10,50,100,200 nmol/L and 1 μmol/L for 24 hours respectively,UVB groups irradiated with UVB of 30,60 and 90 mJ/cm2 respectively,Se-Met + UVB groups treated with Se-Met at concentrations of 1,10,50,100,200 nmol/L and 1 μmol/L for 24 hours firstly,then irradiated with UVB of 30,60 and 90 mJ/cm2 respectively.Subsequently,methyl thiazolyl tetrazolium (MTT) assay was performed to estimate cellular proliferative activity,flow cytometry to detect cell apoptosis,colorimetry to evaluate superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities and to determine malondialdehyde (MDA) levels.Statistical analysis was carried out by using factorial design analysis of variance (ANOVA),one-way ANOVA and least significant difference (LSD) test.Results Factorial design ANOVA showed that UVB radiation had an inhibitory effect on the proliferative activity of HaCaT cells (F =128.04,P ＜ 0.05),which significantly decreased along with the increase of UVB doses,with significant differences between the three UVB groups (P ＜ 0.05).Se-Met pretreatment also affected cellular proliferative activity (F =5.95,P ＜ 0.05),which was significantly increased in Se-Met (10 nmol/L-1 μmol/L) + UVB groups compared with the UVB groups at corresponding doses (all P ＜ 0.05).There was no significant interaction effect on cellular proliferative activity between UVB radiation and Se-Met pretreatment (F =1.65,P ＞ 0.05).The apoptosis rate of HaCaT cells in the 30-mJ/cm2 UVB group was 31.9％ ± 2.67％,significantly higher than that in the normal control group (4.1％ ± 0.67％,P＜ 0.05) and in the 10-,50-,100-,200-nmol/L and 1-μmol/L Se-Met + 30-mJ/cm2 UVB groups (21.9％ ± 3.72％,17.2％ ± 1.67％,4.6％ ±-0.85％,7.5％ ± 1.86％ and 13.5％ ± 1.95％ respectively,all P ＜ 0.05).Similarly,SOD and GSH-Px activities were significantly weaker (both P ＜ 0.05),while MDA levels were higher (all P ＜ 0.05) in the 30-mJ/cm2 UVB group than in the normal control group;however,there was a significant increase in SOD and GSH-Px activities but a decrease in MDA levels in the Se-Met (10 nmol/L-1 μmol/L) + 30-mJ/cm2 UVB groups compared with the 30-mJ/cm2 UVB group (all P ＜ 0.05).Conclusions Se-Met can reduce UVB-induced oxidative damage to HaCaT cells,likely by enhancing antioxidase activity and decreasing oxygen radicals.
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Objective To study the role of D‐lactate gradient across the lung in the rapid diagnosis of pneumonia and evalua‐tion of therapeutic efficacy .Methods Patients were divided into pneumonia group (n=46) and non‐pneumonia group (n=28) in ICU .D‐lactate gradient across the lung were calculated by the difference between arterial and mixed‐venous D‐lactate concentrations before the treatment ,after 3 and 7 days of treatment .Serum procalcitonin (PCT) ,Oxygenation index ,the lung injury score (LIS) and clinical pulmonary infection score(CPIS) were recorded at the same time .Results The mean D‐lactate gradient across the lung in pneumonia group was significantly higher than that in non‐pneumonia group[(163 .84 ± 10 .72) ng/mL vs .(30 .33 ± 7 .25) ng/mL ,P<0 .01) ]before treatment .Using a cut‐off value of 106 .11 ng/mL ,D‐lactate gradient across the lung′s sensitivity for di‐agnosis pneumonia was 90 .7% and its′specificity was 75 .5% .D‐lactate gradient across the lung correlated with LIS (r= 0 .554 , P<0 .01) and CPIS(r=0 .543 ,P<0 .01) .Conclusion D‐lactate gradient across the lung correlates with lung injury and pulmonary infection positively and may be a potential biomarker for rapid diagnosis of pneumonia .
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AIM: To investigate the protective effect of sodium selenite ( Na2 SeO3 ) on human keratinocytes under ultraviolet-B (UVB) irradiation.METHODS: The cultured HaCaT cells were divided into 4 groups: (1) normal control group;(2) Na2 SeO3 group:pretreated with Na2 SeO3 at doses of 10 nmol/L, 50 nmol/L, 100 nmol/L, 200 nmol/L and 1 μmol/L for 24 h;(3) UVB group: irradiated with UVB at doses of 300, 600 and 900 J/m2; (4) Na2SeO3 +UVB group:after pretreated with Na2SeO3 for 24 h, irradiated with UVB at doses of 300, 600 and 900 J/m2.The cell pro-liferation was detected by MTT assay .The apoptotic rates of HaCaT cells treated with UVB at dose of 300 J/m2 were as-sessed by flow cytometry .RESULTS:Compared with normal control group , the cell proliferation activity in UVB group de-creased significantly ( P<0.05 ) .The cell activity was inversely correlated with the irradiation intensity .No significant difference of the cell activity between Na 2 SeO3 group and normal control group was observed .The cell proliferation in Na2SeO3 +UVB group was higher than that in UVB group significantly (P<0.05).Na2SeO3 at concentration of 100 nmol/L showed the strongest activity to promote cell proliferation .After 300 J/m2 UVB irradiation, the apoptotic rate in Na2SeO3+UVB group decreased significantly ( P<0.05) compared with UVB group .The inhibitory effect of Na 2 SeO3 at concentra-tion of 100 nmol/L on apoptosis was the strongest .CONCLUSION: The damage of human keratinocytes by UVB irradia-tion is in a dose-dependent manner .The photoprotection performance of Na 2 SeO3 reduces the damage of human keratino-cytes induced by UVB irradiation .
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Objective To investigate the impact of Ang-1 on the septic mice′pulmonary vascular endothelial barrier function and VE-cadherin and its mechanism. Methods 80 BALB/c mice were randomly divided into NS, LPS, LPS+Ang-1, LPS+Ang-1+ Ly and Ang-1 groups (n = 16). Measure VE-cadherin, Ang-2 levels in plasma and lung permeability index (LPI).Test the total VE-cadherin of lung and the phosphorylation of VE-cadherin expression. Results Plasma Ang-2 was higher compared with NS group(P<0.01) except Ang-1 group. In LPS+Ang-1 group and LPS+Ang-1+Ly group, plasma Ang-2 was lower compared with LPS group (P <0.05). In LPS+Ang-1+Ly group, plasma Ang-2 was higher compared with LPS+Ang-1 group (P<0.01). LPI, plasma VE-cadherin and lung phosphorylation of VE-cadherin were the same with the trends of the plasma Ang-2 , but the lung total VE-cadherin showed the opposite tendency. Conclusion Through the PI3K/Akt signal transduction pathway , Ang-1 may regulate septic mice′VE-cadherin , hence the pulmonary vascular endothelial barrier function improved.
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Objective To construct a bicistronic expression vector containing HPV type 6b L1 gene, to express the recombinant vector in mammalian cells, and to establish a cell strain stably expressing HPV6b L1 gene. Methods After endonuclease digestion and purification, the gene fragment of HPV6b L1 was cloned into the eukaryotic expression vector pIRES2-enhanced green fluorescent protein (EGFP). The identification of the recombinant was realized via endonuclease digestion and sequence analysis. Then, the recombinant plasmid pIRES2-HPV6bLl-EGFP was transfected into NIH3T3 (a mouse embryonic fibroblast cell line) cells. Subsequently, the expression of EGFP was observed by fluorescent inverted microscopy, and HPV6b L1 mRNA expression by reverse transcription (RT)-PCR. Results The recombinant plasmid pIRES2-HPV6bLl-EGFP was successfully constructed, transfected into N1H3T3 cells, and selected by G418. The expression of EGFP was seen under an inverted fluorescence microscoy. RT-PCR proved the expression of HPV6b LI mRNA in transfected cells. Conclusions The recombinant plasmid pIRES2-HPV6bLl-EGFP was successfully constructed and transfected into NIH3T3 cells. Inverted fluorescent microscopy and RT-PCR confirmed the successful expression of HPV6b L1 in NIH3T3 cells.
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Objective To investigate the effects of intense pulsed light (IPL) irradiation on the content of collagen fibers, elastic fibers and hyaluronic acid in Kunming mouse skin. Methods The dorsal skin of mice was divided into two areas: the right area was irradiated with IPL, and the left remaining unirradiated served as the control. Skin specimens were taken from the back of mice on day 1, 3, week 1, 2, 4 and 8 after the irradiation and subjected to staining with HE, sirius red and Gomori aldehyde-fuchsin for examinations of histological changes, type Ⅰ and Ⅲ collagen fibers and elastic fibers. The hydroxyproline and hyaluronic acid content in skin tissues of mice was determined with ultraviolet spectrophotometry and radioimmunoassay respectively. Results After irradiation, a significant increase was observed in dermal thickness on week 2 (t =4.623, P＜ 0.05), 4, and 8 (t = 3.904, P＜ 0.05), in type Ⅲ collagen fiber (t = 5.129, P＜ 0.05) on week 1,in type Ⅰ and Ⅲ collagen fibers on week 2, 4 and 8 (both P＜ 0.05), in elastic fibers from week 2 to 8 (P ＜0.05), and in hydroxyproline content from week 1 to 8 (all P ＜ 0.05) in the skin of mice compared with unirradiated mice. In detail, dermal thickness increased by 18.71% on week 4, and type Ⅲ collagen fiber by 40.54% in irradiated mice compared with unirradiated mice. Further more, the hyaluronic acid content was elevated from day 1 to 3, but gradually declined from week 1 to 8, and remained statistically higher from day 1 to week 8 (P ＜ 0.05) in irradiated mice compared to unirradiated mice. Conclusion IPL irradiation could induce an increase in the content of collagen fiber, elastic fiber and hyaluronic acid in the dorsal skin of mice.
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Objective To investigate the expressions of survivin, cyclooxyenase-2 (COX-2) and vascular endothelial growth factor (VEGF) and their relationship with angiogenesis in condyloma acuminatum (CA) tissues. Methods Immunohistochemistry using PowerVision staining kit was performed to detect the expression of survivin, COX-2 and VEGF protein in 60 CA tissue samples from patients and 21 normal skin samples from the foreskin of human controls. At the same time, the microvessel density was determined in CA tissues by staining blood vessel endothelium with anti-CD105 monoclonal antibody. Results The positivity rate of survivin and COX-2 expression was 56.67% and 63.33%, in CA tissues, 9.52% and 0 in normal skin tissues, respectively. There was a significant difference between the two groups of tissue samples in the positivity rate and intensity of survivin and COX-2 expression (all P < 0.05). VEGF was expressed in all of the CA tissues and normal skin tissues, while the intensity of VEGF expression was statistically different between the two groups of tissue samples (P < 0.05). The MVD was 16.38 ± 5.46 and 0.62 ± 0.44 in CA tissues and normal skin tissues, respectively (P < 0.05). There was a significant positive correlation between the expressions of survivin, COX-2 and VEGF, as well as between MVD and the expressions of survivin and COX-2 in CA tissues. Conclusion The expression levels of survivin, COX-2 and VEGF are significantly higher in CA tissues than in normal skin tissues.
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Objective To assess the mutation in exon 8 of C1 esterase inhibitor(C1INH)gene in a patient with hereditary angioedema(HAE).Methods Genomic DNA was extracted from a female patient with HAE as well as her mother and a normal human control.The fragment of exon 8 of C1INH gene was amplified by PCR and inserted into plasmid carrier pUC19 with the help of ligase.Then,the recombinant plasmid was transformed into competent cells of E coli TG1 strains.After culture of positive transformant,plasmid DNA Was extracted and subjected to sequencing.SDS-PAGE and We:stem blot were performed on the sera of the patient to detect the concentration and function of C1INH protein.Results An A1677G mutation at exon 8 of C1INH gene.which resulted in a substitution of isoleucine to valine at codon 440,Was found in the patient who SUfiered from HAE type I.Additionally.SDS-PAGE and Western blot revealed that the molecular weight of C1INH protein was 96 000.but not 105 000 observed in noHnal human control.Conclusion The newly identified mutation 1440V.which is located at P4 residue of reactive center loop in C1INH.may result in conformational alteration of C1INH.
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Objective: To investigate the influence of Xuebijing injection (血必净注射液) on hemodynamics of dogs with endotoxic shock and its mechanism. Methods: Endotoxic shock was induced by intravenous infusion of lipopolysaccharide of E.coli. O55:B5 to dogs, and the 24 dogs were randomly divided into control group and Xuebijing injection group. Xuebijing injection was administered in the Xuebijing injection group. The hemodynamic parameters and plasma lactate levels were monitored. Results: After the administration of endotoxin, the mean arterial pressure (MAP) decreased significantly, while the mean pulmonary artery pressure (MPAP) and cardiac index (CI) increased markedly in both groups (all P0.05). Moreover, plasma lactate level was lower in Xuebijing injection group than that in control group (P
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Objectives To investigate the expression of matrix metalloproteinase-9 (MMP-9) and its tissue inhibitor (TIMP-1) in cutaneous squamous cell carcinomas (SCC) and the relationship between the expression and invasive growth and metastasis of SCC. Methods Immunohistochemical method of streptavidin-peroxidase (SP) was used to detect the expression of MMP-9 and TIMP-1 proteins on the paraffin embedded sections of 65 patients with cutaneous SCC and 5 cases of normal epidermis. The immunohistochemical results were analyzed quantitatively using an image analysis system. Results MMP-9 and TIMP-1 proteins were diffusely expressed on the tumor nests and the mesenchymal cells around the nests, while in normal epidermis MMP-9 and TIMP-1 were weakly positive. The expression level of MMP-9 protein and the ratio of MMP-9/TIMP-1 were higher in aggressive SCC group than those SCC in situ group (t = 2.33 and 2.36, respectively, P
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Objective To explore the roles of Bcl-2, Bax, Bcl-xl proteins in the carcinogenesis of basal cell carcinoma(BCC). Methods Forty-six BCC cases were divided into 2 groups (21 in non-aggressive BCC group and 26 in aggressive BCC group) according to their pathological features. Bcl-2, Bax and Bcl-xl were evaluated in formalin fixed, paraffin embedded specimens by immunohistochemical methods and analyzed by image-analysis methods to obtain average optical density (AOD) of each protein for each case. The AOD data of 3 proteins and Bcl-2/Bax ratios were then compared between the 2 BCC groups. Results The level of Bcl-2 expression and Bcl-2/Bax ratio of non-aggressive BCC group were higher than those of aggressive group,(P=0.005 and 0.044, respectively). For the expression level of Bcl-xl and Bax, there was no statistically difference between the two groups (P=0.097 and 0.979, respectively). Conclusion Bcl-2, Bax,Bcl-xl proteins might participate in the carcinogenesis and development of BCC.