OBJECTIVETo examine the expression of matrix metalloproteinases (MMP)-2, -9, tissue inhibitor of matrix metalloproteinase (TIMP)-1 and hs-CRP, and their relationship with coronary artery in children with Kawasaki disease.

METHODSOne hundred and fifty-one children with Kawasaki disease (111 cases with coronary artery damage and 40 cases without) and 60 healthy children were enrolled. The expression of MMP-2, MMP-9 and TIMP-1 was detected using ELISA, and the hs-CRP concentration was measured using the endpoint nephelometry.

RESULTSThere were significant differences in the level of MMP-2, MMP-9 and hs-CRP between the patients with or without coronary artery damage and the healthy children (p<0.05). The levels of MMP-2, MMP-9 and hs-CRP were the highest in the cardiovascular damage group (p<0.05). There were positive correlations between MMP-2, MMP-9 and TIMP-1 in children with Kawasaki disease.

CONCLUSIONSMMP-2, MMP-9, TIMP-1 and hs-CRP may play important roles in the development of Kawasaki disease. The combined measurement of MMP-2, MMP-9 and hs-CRP may be useful in the evaluation of the severity in children with Kawasaki disease.

C-Reactive Protein ; analysis ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Matrix Metalloproteinase 2 ; blood ; Matrix Metalloproteinase 9 ; blood ; Mucocutaneous Lymph Node Syndrome ; blood ; etiology ; Tissue Inhibitor of Metalloproteinase-1 ; blood

DNA, Complementary ; chemistry ; Humans ; Leukocytes ; enzymology ; Lung Neoplasms ; enzymology ; pathology ; Matrix Metalloproteinase 2 ; blood ; genetics ; Matrix Metalloproteinase 9 ; blood ; genetics ; Neoplasm Invasiveness ; Neoplasm Metastasis ; RNA, Messenger ; analysis ; Tissue Inhibitor of Metalloproteinase-1 ; blood ; genetics

OBJECTIVETo investigate remodeling characteristics of coronary lesions in patients with acute coronary syndromes (ACS) versus stable angina pectoris (SA) using intravascular ultrasound (IVUS), and to explore the relationship between arterial remodeling and clinical presentation or matrix metalloproteinase (MMPs) or hyper-sensitive C-reactive protein (hs-CRP).

METHODSWe studied culprit lesions of 38 patients with ACS and 18 patients with SA using IVUS before coronary intervention. The lesion site and a proximal or distal reference site including the external elastic membrane (EEM) area and lumen area were analyzed. Plaque area and remodeling index (RI) were calculated, and directions of arterial remodeling were determined. Positive remodeling was defined as RI > 1.05 and negative remodeling as RI < 0.95. We analyzed the culprit lesion qualitatively, identified high risk plaque and compared them in each group. The blood level of MMP-2, MMP-9 and hs-CRP in each group were also determined.

RESULTSThe plaque area at culprit lesions in patients with ACS was significantly larger (11.94 +/- 4.90 versus 9.17 +/- 3.36 mm2; P = 0.035), and also the RI in ACS group was significantly greater than that of patients with SA (0.972 +/- 0.222 versus 0.796 +/- 0.130; P = 0.003). The distribution of remodeling in these two groups was different: positive remodeling was more frequent in ACS group than in SA group (34.2% versus 5.6%, P = 0.047), whereas negative remodeling was more frequent in SA group (52.6% versus 88.9%, P = 0.003). There was higher incidence of high risk plaque in ACS group compared to SA (76.3% versus 50.0%, P = 0.040). The level of serum MMP-2 in ACS group was higher than that of SA group (250.65 +/- 47.97 microg/L versus 214.21 +/- 47.20 microg/L, P = 0.029). The same applied for plasma MMP-9 (84.26 +/- 9.78 microg/L versus 68.46 +/- 22.82 microg/L, P = 0.038) and serum hs-CRP (3.62 +/- 3.37 mg/L versus 1.48 +/- 1.52 mg/L, P = 0.041).

CONCLUSIONSPositive remodeling, larger plaque area and higher incidence of high risk plaque are associated with ACS, whereas negative remodeling is more common in patients with SA. This association between the extent of remodeling and clinical presentation may reflect a greater tendency that plaques with positive remodeling can cause ACS. The change of level of MMP-2, MMP-9 and hs-CRP in ACS patients may be helpful in investigating vulnerable plaques.

Adult ; Aged ; C-Reactive Protein ; analysis ; Coronary Disease ; blood ; pathology ; Coronary Vessels ; diagnostic imaging ; pathology ; Female ; Humans ; Male ; Matrix Metalloproteinase 2 ; blood ; Matrix Metalloproteinase 9 ; blood ; Middle Aged ; Ultrasonography, Interventional

OBJECTIVETo study the expression of gelatinases, including matrix metalloproteinase-9 (MMP-9, gelatinase B) and matrix metalloproteinase-2 (MMP-2, gelatinase A), in CD(34)(+) cells and leukemic cell lines, and explore the significance of gelatinase in migrating and homing capacity of CD(34)(+) cells, as well as the role of gelatinase in leukemia pathogenesis.

METHODSCD(34)(+) cells were isolated from umbilical cord blood and normal bone marrow by Mini MACS system. By zymogram analysis, MMP-2 and MMP-9 were detected in the serum free condition medium of CD(34)(+) cells and cell lines.

RESULTSOne brilliant band with molecular weight of 92 x 10(3) was detected in condition medium of cord blood CD(34)(+) cells. No band was detected in condition medium of bone marrow CD(34)(+) cells. Brilliant bands with molecular weight of 92 x 10(3) and 72 x 10(3) were detected in the condition medium of U937, KG-1a and HL-60 cell lines, but not in that of HEL, Namalva, CEM, K562 and LCL-H cell lines. In the condition media of J6-1 and J6-2 cells only the 92 x 10(3) band was detected.

CONCLUSIONSCord blood CD(34)(+) cells produced MMP-9, but bone marrow CD(34)(+) cells did not, partly explains the fact that cord blood CD(34)(+) cells possessed higher migrating capacity in comparison with bone marrow CD(34)(+) cells. The expression of MMP-9 and MMP-2 in leukemic cell lines varied.

Antigens, CD34 ; analysis ; Culture Media, Conditioned ; metabolism ; Electrophoresis, Polyacrylamide Gel ; Fetal Blood ; cytology ; enzymology ; immunology ; HL-60 Cells ; Humans ; K562 Cells ; Leukemia ; enzymology ; pathology ; Matrix Metalloproteinase 2 ; biosynthesis ; Matrix Metalloproteinase 9 ; biosynthesis ; Tumor Cells, Cultured ; U937 Cells

The levels of tartrate resistant acid phosphatase (TRAP), matrix metalloproteinase-2 (MMP-2), and tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) in synovial fluid (SF) and serum in cases of canine osteoarthritis (OA) were measured. OA was induced by a surgically-created medial patellar luxation in the left stifle of 24 dogs. SF and blood samples were collected at 1.5- and 3-month intervals, respectively. Every 3 months, one dog was euthanatized to collect tissue samples from both stifles. TRAP levels in SF and serum were measured using a spectrophotometer, and TRAP-positive cells in joint tissues were identified by enzyme histochemistry. MMP-2 and TIMP-2 in SF and serum were detected by Western blotting and ELISA, respectively. TRAP in SF from the stifles and serum was significantly increased (p < 0.05) after 3 months. TIMP-2 in SF and serum was significantly decreased (p < 0.05), whereas MMP-2 in SF was significantly increased (p < 0.05) during the progression of OA. Histochemistry revealed an increased number of TRAP-positive cells in tissues from OA-affected joints. Assays measuring TRAP, MMP-2, and TIMP-2 in SF and serum, and methods that detect increased numbers of TRAP-positive cells in the joint tissues can play an important role in identifying the early phases of degenerative changes in canine joint components.
Acid Phosphatase/analysis/blood ; Animals ; Arthritis, Experimental/enzymology/etiology/veterinary ; Biological Markers/*analysis/*blood ; Blotting, Western/veterinary ; Dislocations/complications/*veterinary ; Dog Diseases/*enzymology/etiology ; Dogs ; Enzyme-Linked Immunosorbent Assay/veterinary ; Female ; Isoenzymes/analysis/blood ; Male ; Matrix Metalloproteinase 2/analysis/blood ; Osteoarthritis/enzymology/etiology/*veterinary ; Spectrophotometry/veterinary ; Stifle/physiopathology ; Synovial Fluid/*enzymology ; Tissue Inhibitor of Metalloproteinase-2/analysis/blood

BACKGROUNDIn multiple myeloma (MM), bone marrow angiogenesis parallels tumour progression and correlates with disease activity. Recent studies have proved resveratrol possesses antiangiogenic activity in vitro and in vivo. In this study, we examined the effects of resveratrol on myeloma cell dependent angiogenesis and the effects of resveratrol on some important angiogenic factors of RPMI 8226 cells.

METHODSRPMI 8226 cells were cocultured with human umbilical vein endothelial cells (HUVECs) to evaluate the effects of myeloma cells on angiogenesis. The RPMI 8226 cells were treated with various concentrations of resveratrol (6.25 - 50.00 micromol/L) for different times (12 - 72 hours). Reverse transcriptase polymerase chain reaction (RT-PCR) was used to assay vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), metalloproteinases (MMP)-2 and MMP-9 mRNA. Gelatin zymography was used to analyze MMP-2 and MMP-9 activity. VEGF and bFGF proteins secreted by the cells in the medium were quantified by enzyme linked immunosorbent assay (ELISA).

RESULTSCell proliferation, migration and differentiation of HUVECs markedly increased by coculture with RPMI 8226 cells. Resveratrol inhibited proliferation, migration and tube formation of HUVECs cocultured with myeloma cells in a dose dependent manner. Treatment of RPMI 8226 cells with resveratrol caused a decrease in MMP-2 and MMP-9 activity. Resveratrol inhibited VEGF and bFGF protein expression in a dose and time dependent manner. Furthermore, decreased levels of VEGF, bFGF, MMP-2 and MMP-9 mRNA from cells treated with various concentrations of resveratrol confirmed its antiangiogenic action at the level of gene expression.

CONCLUSIONSResveratrol inhibits multiple myeloma angiogenesis by regulating expression and secretion of VEGF, bFGF, MMP-2 and MMP-9. Resveratrol may be a potential candidate for the treatment of multiple myeloma.

Angiogenesis Inhibitors ; pharmacology ; Antineoplastic Agents, Phytogenic ; pharmacology ; Cell Line, Tumor ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Fibroblast Growth Factor 2 ; analysis ; genetics ; Humans ; Matrix Metalloproteinase 2 ; analysis ; Matrix Metalloproteinase 9 ; analysis ; Multiple Myeloma ; blood supply ; drug therapy ; pathology ; RNA, Messenger ; analysis ; Stilbenes ; pharmacology ; Vascular Endothelial Growth Factor A ; analysis ; genetics

OBJECTIVESTo explore the role of fibroblasts derived from tumor in the tumor angiogenesis.

METHODSTwo-well co-culture system were used to detect the expression of MMP-9, TGF-beta1, TN and bcl-2 in L929-H22 cells, and their ability of promoting angiogenesis of ECV304 cells and invasion of MDA-MB-231 cells respectively, which were established in our laboratory before. Then their adhesion and the effect of their supernatant on H22 cells proliferation were analysed.

RESULTSCompared with L929 cells, the adhesion potential of L929-H22 cells increased (F>or=104.32, P<0.001), with the higher level of expression of MMP-9, bcl-2, TN, and TGF-beta1 in L929-H22 cells in creased (t>or=3.3055, P<0.01). L929-H22 and L929 cells enhanced the invasiveness of human mammary cancer MDA-MB-231 cells through artificial basement membrane (Matrigel) 1.21 and 0.48 times respectively (F=266.3, P<0.001). L929-H22 cells induced morphogenesis of ECV304 cells. L929-H22 stimulated endothelial cells to form more and longer tubes than L929 did (F>or=23.75, P<0.01). 25% CM of L929-H22 cells stimulated the growth of H22 cells (F=266.30, P<0.05).

CONCLUSIONThe results suggested that fibroblasts in tumors secrete more growth factors and angiogenic factors to promote the angiogenesis and invasion of solid tumors.

Animals ; Cell Line, Tumor ; Fibroblasts ; physiology ; Humans ; Matrix Metalloproteinase 9 ; analysis ; Mice ; Neoplasm Invasiveness ; Neoplasms ; blood supply ; Neovascularization, Pathologic ; etiology ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Transforming Growth Factor beta ; analysis

BACKGROUNDExcessive deposition of extracellular matrix (ECM) in the kidney is the hallmark of diabetic nephropathy. Increased matrix synthesis has been well documented but the effects of diabetes on degradative pathways, particularly in the in vivo setting. The renal protective effect of these pathways on matrix accumulation has not been fully elucidated. The present study was undertaken to investigate the activity of matrix metalloproteinase-2 (MMP-2), the expression of MMP-2 and tissue inhibitor of metalloproteinase-2 (TIMP-2) in kidney tissues of diabetic rats, and to explore the degradative pathway of type IV collagen (IV-C) and the renal protective effects of ACE inhibition-benazepril.

METHODSTwenty-four healthy male Wistar rats were divided randomly into normal control group (NC group), untreated diabetes mellitus group (DM group), and diabetes mellitus group treated with benazepril (DL group). The rat model of diabetes mellitus was induced by intraperitoneal injection of streptozocin (60 mg/kg). After the establishment of DM model, benazepril (10 mg.kg(-1).d(-1)) was given to the DL group for 12 weeks, and the same volume of water was given to the other two groups. At the end of 12 weeks, renal function was evaluated with 24-hour urinary protein (Upro), clearance of creatinine (Ccr), and blood urea nitrogen (BUN). MMP-2 activity was determined by gelatin zymography. The levels of MMP-2, TIMP-2 and collagen IV (IV-C) protein in the kidney tissue were assessed by immunohistochemistry. The gene expression of MMP-2 and TIMP-2 was measured by reverse transcription polymerase chain reaction (RT-PCR).

RESULTSThe levels of BUN, Upro and Ccr in the DM group were higher than those in the NC group. In the DM group, the mRNA, enzymatic activity and proteins of MMP-2 decreased, but the expressions of IV-C and TIMP-2 increased. All diabetes-associated changes in renal function and MMP/TIMP were attenuated after benazepril treatment with reduced IV-C accumulation.

CONCLUSIONSThe changes of MMP-2 and TIMP-2 expressions in kidney tissues of diabetes rats may contribute to the occurrence and progression of diabetic nephropathy. Benazepril could exert protective effects on diabetic nephropathy, owing to the upregulation of MMP-2 and downregulation of TIMP-2 expressions, which further inhibits the excessive deposition of extracellular matrix in the glomerulus.

Angiotensin-Converting Enzyme Inhibitors ; pharmacology ; Animals ; Benzazepines ; pharmacology ; Blood Glucose ; analysis ; Body Weight ; Collagen Type IV ; metabolism ; Diabetes Mellitus, Experimental ; metabolism ; pathology ; Diabetic Nephropathies ; prevention & control ; Kidney ; drug effects ; metabolism ; Kidney Glomerulus ; pathology ; Male ; Matrix Metalloproteinase 2 ; analysis ; genetics ; Organ Size ; Rats ; Rats, Wistar ; Streptozocin ; Tissue Inhibitor of Metalloproteinase-2 ; analysis ; genetics

This study was designed to evaluate the inhibition effect of lambda-carrageenan oligosaccharides on neovascularization in vitro by chick chorioallantoic membrane (CAM) model and human umbilical vein endothelial cell ( HUVEC). lambda-Carrageenan oligosaccharides caused a dose-dependent decrease of the vascular density of CAM, and adversely affected capillary plexus formation. At a high concentration of 1 mg x mL(-1), this compound inhibited the endothelial cell proliferation, while low concentration of lambda-carrageenan oligosaccharides (< 250 microg x mL(-1)) affected the cell survival slightly (> 95%). Different cytotoxic sensitivity of lambda-carrageenan oligosaccharides in three kinds of cells was observed, of which HUVEC is the most sensitive to this oligosaccharides. The inhibitory action of lambda-carrageenan oligosaccharides on the endothelial cell invasion and migration was also observed at relatively low concentration (150 - 300 microg x mL(-1)) through down-regulation of intracellular matrix metalloproteinases-2 (MMP-2) expression on endothelial cells. Current observations demonstrated that lambda-carrageenan oligosaccharides are potential angiogenesis inhibitor with combined effects of inhibiting invasion, migration and proliferation.
Angiogenesis Inhibitors ; pharmacology ; Animals ; Carrageenan ; pharmacology ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Chick Embryo ; Chorioallantoic Membrane ; blood supply ; cytology ; drug effects ; Endothelial Cells ; drug effects ; Humans ; Matrix Metalloproteinase 2 ; analysis ; Oligosaccharides ; pharmacology

BACKGROUNDPrevious studies have indicated that thrombin (TM) may play a major role in brain edema after intracerebral hemorrhages (ICHs). However, the mechanism of TM-induced brain edema is poorly understood. In this study, we explored the effect of TM on the permeability of the blood brain barrier (BBB) and investigated its possible mechanism, aiming at providing a potential target for brain edema therapy after ICHs.

METHODSTM or TM + cathepsin G (CATG) was stereotaxically injected into the right caudate nucleus of Sprague-Dawley rats in vivo. BBB permeability was measured by Evans-Blue extravasation. Brain water content was determined by the dry-wet weight method. Brain microvascular endothelial cells were then cultured in vitro. After TM or TM + CATG was added to the endothelial cell medium, changes in the morphology of cells were dynamically observed by phase-contrast light microscopy, and the expression of matrix metalloproteinase-2 (MMP-2) protein was measured by immunohistochemical method.

RESULTSBBB permeability increased at 6 hours after a TM injection into the ipsilateral caudate nucleus (P < 0.05), peaked between 24 hours (P < 0.01) and 48 hours (P < 0.05) after the injection, and then declined. Brain water content changed in parallel with the changes in BBB permeability. However, at all time points, BBB permeability and brain water content after a TM + CATG injection were not significantly different from the respective parameters in the control group (P > 0.05). TM induced endothelial cell contraction in vitro in a time-dependent manner and enhanced the expression of MMP-2 protein. After incubation with TM + CATG, cell morphology and MMP-2 expression did not change significantly as compared to the control group (P > 0.05).

CONCLUSIONSIncreased BBB permeability may be one of the mechanisms behind TM-induced cerebral edema. TM induces endothelial cell contraction and promotes MMP-2 expression by activating protease activated receptor-1 (PAR-1), possibly leading to the opening of the BBB.

Animals ; Blood-Brain Barrier ; drug effects ; Body Water ; metabolism ; Brain Edema ; etiology ; Cathepsin G ; Cathepsins ; pharmacology ; Cerebral Hemorrhage ; complications ; Matrix Metalloproteinase 2 ; analysis ; Permeability ; Rats ; Rats, Sprague-Dawley ; Receptor, PAR-1 ; physiology ; Serine Endopeptidases ; Thrombin ; toxicity