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Fang C.,Shandong University | Fang C.,Queen Mary, University of London | Wen G.,Queen Mary, University of London | Wen G.,Guangzhou University | And 7 more authors.
Cardiovascular Research | Year: 2013

AimsGrowing evidence suggests a close association of plaque angiogenesis with atherosclerotic plaque formation and progression, and an important role of matrix metalloproteinase (MMP) in angiogenesis and atherosclerosis. We attempted to investigate the functional involvements of MMP8 in angiogenesis.Methods and resultsKnockdown of MMP8 in human umbilical vein endothelial cells (HuVECs) with MMP-8 shRNA lentivirus resulted in a decrease in in vitro capillary-like network formation, cell proliferation and migration, and impaired its capacity of in vivo angiogenesis. Less nuclear accumulation of β-catenin and lower β-catenin target gene expression levels was observed in the HuVECs expressing lower levels of endogenous MMP8. Knockdown of endogenous MMP8 in HuVECs down-regulated platelet/endothelial cell adhesion molecule-1 (PECAM-1) expression by converting less angiotensin I to II, which is an inducer for PECAM-1 gene expression. Aortic rings isolated from MMP8-/-/apoE -/- mice had less endothelial cell sprouting, and endothelial cells in MMP8-/-/apoE-/- mice had a lower ability to migrate into Matrigel plugs and less capacity of proliferation and angiogenesis. Moreover, immunohistochemical analyses revealed that MMP8 was expressed in microvessels within human atherosclerotic plaques and aneurysm. Finally, analyses of MMP8-/-/apoE-/- and MMP8+/+/ apoE-/- mice fed a Western diet for 12 weeks showed that MMP8-deficient mice had small lesion size and less endothelial cells within atherosclerotic lesions.ConclusionWe demonstrated for the first time that MMP8 plays an important role in angiogenesis in vitro and in vivo. Our findings provide new insights into the molecular mechanisms of plaque angiogenesis and suggest that MMP8 is a potential therapeutic target of cardiovascular diseases. © 2013 Published on behalf of the European Society of Cardiology. All rights reserved. © The Author 2013. Source

Li C.-X.,University of South China | Li C.-X.,Dongguan Institute of Dermatology | Han C.-L.,Dongguan Institute of Dermatology | Zeng K.,University of South China | And 2 more authors.
British Journal of Dermatology | Year: 2014

Background A series of cases of symmetrical acral keratoderma have been described recently in China. However, no studies about its demographic information and epidermal barrier function have been documented. Objectives To describe the clinical manifestation, demographic information and clinicopathological features of 71 cases with symmetrical acral keratoderma. Patients and methods Seventy-one cases with symmetrical acral keratoderma were retrospectively reviewed. Their demographic information, clinical manifestations, histopathology and epidermal barrier function were analysed. Results Among these patients, there were 64 males and seven females, ranging in age from 4 to 53 years with an average age at onset of 27 ± 8·9 years. Clinical manifestation was characterized by brown hyperkeratotic patches over the dorsum of the hands, palms and feet, dorsal digits and wrists, elbows, knees and ankles. The lesions became dramatically whitish with mild swelling immediately after soaking in water and resolved spontaneously in winter. In patients, a moderate increase of transepidermal water loss (TEWL) from 16·16 ± 6·15 to 9·9 ± 4·21 g m -2 h-1 (P = 0·0054) and a moderate decrease of skin hydration from 65·9 ± 5·06 to 42·58 ± 10·73 (P < 0·01) in comparison with the control group were observed. Histopathological examination revealed epidermal hyperkeratosis, acanthosis and papillomatous hyperplasia as well as dermal infiltration with a few lymphocytes. Conclusions Symmetrical acral keratoderma is characterized by symmetry, acra, keratinization and marked seasonal changes. The epidermal barrier function of the skin was negatively affected. What's already known about this topic? A series of cases of symmetrical acral keratoderma have been described recently in China. However, no studies of demographic information and epidermal permeability barrier function have been documented. What does this study add? Symmetrical acral keratoderma is characterized by symmetry, acra, keratinization and marked seasonal changes. The epidermal barrier function of the skin was negatively affected. © 2013 British Association of Dermatologists. Source

Ge C.,Chinese Academy of Sciences | Ge C.,Hefei University of Technology | Luo Q.,Guangzhou Institute of Dermatology | Wang D.,Chinese Academy of Sciences | And 6 more authors.
Analytical Chemistry | Year: 2014

G-quadruplex-forming sequence can be formed through a copper(I) ion (Cu+)-catalyzed click chemistry between azide-and alkyne-modified short G-rich sequences in aqueous solution, eliminating immobilization and washing steps of conventional assays. The source for Cu+ was generated from the reduction of Cu2+ with the reductant of sodium ascorbate. In the presence of hemin and K+, the self-assembly of hemin/G-quadruplex structure has the activity of horseradish peroxidase (HRP), which can catalyze its colorless substrate tetrazmethyl benzidine (TMB) into a colored product. Hence, the concentration of Cu2+ can be evaluated visually for qualitative analysis according to the color change of the solution, and the optical density (OD) value of the resulting solution at 450 nm was also recorded using a microplate reader for quantitative analysis. © 2014 American Chemical Society. Source

Sanquan Z.,Guangzhou Institute of Dermatology
National Medical Journal of China | Year: 2014

Objective: To explore the effects of cell cycle distribution and expression of cell cycle related proteins before and after transglutaminase 1 (TGM1) gene silencing by small interfering RNA (siRNA) in immortalized human keratinocytes (HaCaT cells). Methods: Before and after RNA interference (RNAi) silencing TGM1 gene expression in HaCaT cells, the changes of cell cycle status were detected by flow cytometry. And the expressions of cell cycle related proteins cyclin Dl, cyclin Bl and CDK4 were detected by immunocytochemistry and Western blot. Results: After TGM1 silencing, the HaCaT cells increased in G0/G1 phase (78. 27% ± 1. 83% vs 56. 84 % ± 2. 72% and 57. 19% ± 3. 72%, both P <0. 05) while decreased in S phases (32. 78% ±5. 48% vs 57. 32% ±2. 91% and 55. 71 % ±2. 84%, both P <0. 05) and G2/M phases(3. 66% ±0. 30% vs 9. 39% ±0. 68% and 9. 77% ±0. 52%,both P < 0. 05). Moreover, arrest of cell cycle was induced in S phase. After TGM1 silencing, cell proliferation was significantly inhibited while wild-type TGM1 displayed an accelerated growth rate. Inmmnohistochemistry indicated that cyclin Dl, cyclin Bl and CDK4 had a strongly negative expression after transfecting with TGM1 siRNA. Western blot indicated that a very low endogenous expression of cyclin Dl, cyclin Bl and CDK4 proteins was observed after transfecting with TGM1 siRNA. Conclusions: TGM1 siRNA may block the cell cycle of HaCaT cells. It suggests that TGM1 expression in HaCaT cells are closely related with cell cycle. Source

He Y.,Guangzhou Institute of Dermatology | He Y.,North Dakota State University | Zeng K.,Southern Medical University | Zhang S.,Guangzhou Institute of Dermatology | And 4 more authors.
Biosensors and Bioelectronics | Year: 2012

Here, we describe a simple and sensitive approach for visual detection of gene mutations based on isothermal strand-displacement polymerase reactions (ISDPR) and lateral flow strip (LFS). The concept was first demonstrated by detecting the R156H-mutant gene of keratin 10 in Epidermolytic hyperkeratosis (EHK). In the presence of biotin-modified hairpin DNA and digoxin-modified primer, the R156H-mutant DNA triggered the ISDPR to produce numerous digoxin- and biotin-attached duplex DNA products. The product was detected on the LFS through dual immunoreactions (anti-digoxin antibody on the gold nanoparticle (Au-NP) and digoxin on the duplex, anti-biotin antibody on the LFS test zone and biotin on the duplex). The accumulation of Au-NPs produced the characteristic red band, enabling visual detection of the mutant gene without instrumentation. After systematic optimization of the ISDPR experimental conditions and the parameters of the assay, the current approach was capable of detecting as low as 1-fM R156H-mutant DNA within 75. min without instrumentation. Differentiation of R156H- and R156C-mutant DNA on the R156 mutation site was realized by using fluorescein- and biotin-modified hairpin probes in the ISDPR process. The approach thus provides a simple, sensitive, and low-cost tool for the detection of gene mutations. © 2011 Elsevier B.V. Source

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