Cell Line, Tumor ; Humans ; Matrix Metalloproteinase 2 ; analysis ; Matrix Metalloproteinase 9 ; analysis ; Melanoma ; enzymology ; Skin Neoplasms ; enzymology

OBJECTIVE: To investigate the correlation between matrix metalloproteinase (MMP)-2/MMP-9 levels and childhood caries, and the saliva levels of MMP-2/MMP-9 among healthy children and those with different degrees of dental caries, both before and after treatment. METHODS: In the study, 368 children aged 3 to 5 years were separated into three groups: severe caries group (112 children), mild caries group (98 children) and caries free group (158 children). The children with severe caries were included in treatment group (83 children) after accepting a comprehensive treatment of caries. MMP-2 and MMP-9 levels were determined by enzyme-linked immunosorbent assay (ELISA) and the data were analyzed by the Statistics Package for Social Science (SPSS 13.0). The differences among severe caries group, mild caries group and caries free group were analyzed by SNK-q (Student Newman Keuls). The severe caries group and treatment group were compared by paired t test. The differences between each group were statistically analyzed. RESULTS: There was no significant difference of the age and gender composition among severe caries group, mild caries group, caries free group and treatment group. The MMP-2 level of severe caries group [(141.3±32.5) μg/L] was higher than those of mild caries group [(107.5±21.3) μg/L] and caries free group [(102.8±18.5) μg/L] (P<0.05). There was no significant difference between mild caries and caries free group (P>0.05). After analysis of 83 children in the treatment group, the level of MMP-2 [(120.1±24.8) μg/L] was lower than before [(144.6±30.3) μg/L] (P<0.05), but was higher than that of caries free group (P<0.05). The MMP-9 levels of severe caries group [(445.8±68.1) μg/L] and mild caries group [(428.6±59.2) μg/L] were higher than that of caries free group [(385.4±60.6) μg/L] (P<0.05), but the difference between severe caries group and mild caries group was not significant (P>0.05). After analysis of 83 children in the treatment group, the alteration of MMP-9 [(432.2±64.7) μg/L] was not significant either (P>0.05). CONCLUSION The saliva levels of MMP-2 and MMP-9 in children with severe caries were higher than those in caries free children, even if the treatment was implemented, which suggests that the MMP-2 and MMP-9 in saliva might be related to the caries in children.
Child, Preschool ; Dental Caries/enzymology* ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Male ; Matrix Metalloproteinase 2/analysis* ; Matrix Metalloproteinase 9/analysis* ; Matrix Metalloproteinases ; Saliva/chemistry*

OBJECTIVE: To investigate the influence of transforming growth factor-alpha (TGF-alpha) on the expression of Matrix metalloproteinase-2 (MMP-2) and Matrix metalloproteinase-9 (MMP-9) mRNA in mouse embryos. MATERIALS AND METHOD: Eight-cell stage mouse embryos were cultured for 48hours with TGF-alpha at concentrations of 1, 10 and 100 ng/ml. Embryos not treated with TGF-alpha served as control. Reverse transcription-polymerase chain reaction (RT-PCR) has been used to examine the expression of MMP-2 and MMP-9 mRNA in developed blastocysts. Following reverse transcription, strategically designed nested primers, optimized for specificity, were used for amplification from the cDNA equivalent of a single embryo. The products were then verified by restriction enzyme digestion and sequence analysis. Results were analyzed with analysis of variance (ANOVA) and statistical significance was defined as p<0.05. RESULTS: The relative quantities (relative volume x intensity) of MMP-2 mRNA expressed in embryos of 10 and 100 ng/m of TGF-alpha treatment groups were significantly increased than in the control and 1 ng/ml of TGF-alpha treatment group (67.2+/-7.5 and 77.4+/-11.6 vs. 38.6+/-4.5 and 43.4+/-6.1, p<0.001). The relative quantities of MMP-9 mRNA of 100 ng/ml TGF-alpha treatment group was significantly increased than in the control and 1 ng/ml TGF-alpha treatment groups (67.6+/-6.5 vs. 36.6+/-14.2 and 40.2+/-11.3, p<0.001, p<0.01, respectively). CONCLUSION: This study suggests that TGF-alpha itself may induce the expression of MMP-2 and 9 mRNA in mouse embryos.
Animals ; Blastocyst ; Digestion ; DNA, Complementary ; Embryonic Structures* ; Matrix Metalloproteinase 2* ; Matrix Metalloproteinase 9* ; Mice* ; Reverse Transcription ; RNA, Messenger* ; Sensitivity and Specificity ; Sequence Analysis ; Transforming Growth Factor alpha

BACKGROUNDMatrix metalloproteinases (MMPs) are the major regulators of collagen degradation involved in the pathogenesis of several diseases of the heart. The purpose of this study was to investigate the dynamic changes in myocardial MMP activity in mice with viral myocarditis (VM), the relationship between MMP activity and both cardiac function and the quantity of myocardial collagen, and the role MMPs playing in the pathological lesions of VM.

METHODSSixty-five six-week-old male DBA/2 mice were divided into two groups. Mice in the infected group (n = 50) were inoculated intraperitoneally with 0.14 ml of Coxsackievirus B3 (CVB3, Nancy strain). Control mice (n = 15) were inoculated intraperitoneally with 0.14 ml of Eagle's medium. Eight infected mice and three control mice were sacrificed on each of days 3, 7, 10, 21 and 30 after inoculation. MMP activity was measured on an SDS-PAGE substrate gel embedded with type I gelatin (zymography). Echocardiographic studies were performed under anesthesia with 3% chloralhydrate administered intraperitoneally (0.01 ml/g - 0.015 ml/g). Cardiac systolic function indices, such as peak velocity of the aorta (Vp), flow velocity integral of the aorta (Vi), ejection fraction (EF), and fractional shortening (FS) were determined by echocardiography. Histological cross sections of the hearts were stained with hematoxylin-eosin and myocardial histopathological scores were determined under an optical microscope. The amount of myocardial collagen was measured by means of hydroxyproline quantification.

RESULTSIn virus-infected mice, both MMP-2 and MMP-9 activities were significantly higher than in control mice, reaching a peak on day 10 (P < 0.01). On day 10, cardiac systolic function indices (EF, FS, Vp, and Vi) were all significantly lower compared both to other stages following viral inoculation and to the control group (P < 0.05). In the acute stage, the amount of myocardial collagen in mice with VM was not significantly different from normal control mice (P > 0.05). However, the amount of myocardial collagen in infected mice at the recovery stage (on days 21 and 30) was significantly greater than those of the control mice. MMP-2 and MMP-9 activities positively correlated with myocardial histopathological scores (r = 0.801, 0.821, P < 0.01) and negatively correlated with Vp (r = -0.649, -0.683, P < 0.01) and Vi (r = -0.711, -0.755, P < 0.01). However, Vp negatively correlated with myocardial histopathological scores (r = -0.756, P < 0.01).

CONCLUSIONSIn mice with VM, the activities of myocardial MMP-2 and MMP-9 increase significantly during the acute stage, and the total quantity of myocardial collagen increases by the time of recovery. These changes are associated with myocardial interstition remodeling and cardiac dysfunction. MMP activity is an important reference marker for myocardial pathological lesions and can be used to evaluate the severity of myocardial interstitial damage and cardiac dysfunction.

Animals ; Collagen ; analysis ; Enterovirus B, Human ; Enterovirus Infections ; enzymology ; pathology ; physiopathology ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Mice ; Mice, Inbred DBA ; Myocarditis ; enzymology ; pathology ; physiopathology

OBJECTIVETo study the role of the synthesis and degradation of collagen at the transcription level during liver fibrogenesis due to schistosomiasis japonica in rabbits.

METHODSNew Zealand rabbits challenged by cercariae of Schistosoma japonicum (S. japonicum) were served as animal models for liver fibrosis. Liver specimens were collected through operations at 4, 6, 8, 10, 12, 16, 20, 24 and 28 wks after challenge. Type I collagen, type III collagen, type IV collagen, MMP-1 and MMP-9 mRNA levels of liver tissue were detected by RT-PCR + Dot blot. The size of egg granulomas and the degree of liver fibrosis were measured by histopathological examinations.

RESULTSType I collagen, type III collagen, type IV collagen, MMP-1 and MMP-9 mRNA levels increased simultaneously in the early stage after challenge. Most of them reached their peak at 10 weeks, and compared with normal controls, type I collagen, type III collagen, type IV collagen, MMP-1 and MMP-9 mRNA levels increased by 12.0-, 11.0-, 6.6-, 10.0- and 11.0-fold, respectively, coinciding with the change of egg granulomas, i.e., the change in the inflammatory process. Then both collagen and collagenase mRNA levels decreased. Type I, III and IV collagen mRNA levels declined to 2-fold to 3-fold as compared with normal controls (P < 0.05), while MMP-1 and MMP-9 mRNA levels declined close to normal levels (P > 0.05) at 28 wks. This study shows that the synthesis and degradation of collagen keep a dynamic balance at the early stage of schistosomiasis japonica challenge, while at the later stages the quantity of collagen synthesis was higher than that of collagen degradation.

CONCLUSIONSIt was confirmed at transcription level that when the quantity of collagen synthesis was higher than that of collagen degradation liver fibrogenesis may be resulted in.

Animals ; Collagen ; metabolism ; Liver Cirrhosis, Experimental ; metabolism ; Matrix Metalloproteinase 1 ; genetics ; Matrix Metalloproteinase 9 ; genetics ; RNA, Messenger ; analysis ; Rabbits ; Schistosomiasis japonica ; metabolism ; Transcription, Genetic

BACKGROUNDTissue inhibitor of matrix metalloproteinase-1 (TIMP-1) is related to the aging of many organs, but few data are available on the change of TIMP-1 in liver aging. The purpose of this study was to investigate the expression and role of TIMP-1, matrix metalloproteinase-2 (MMP-2) and MMP-9 in the process of natural aging in the livers of normal and transgenic mice, and to detect the effects of TIMP-1 on oxidative level and anti-oxidative ability of the livers of transgenic young mice.

METHODSNormal and transgenic mice were divided into 3 groups according to their age: 3-month-old group (n = 5), 12-month-old group (n = 5) and 24-month-old group (n = 5). Histopathological changes of the liver were observed after HE and Masson staining. The messenger RNA (mRNA) levels of TIMP-1, MMP-2 and MMP-9 were determined by semi-quantitative reverse transcriptional polymerase chain reaction; protein expression was measured by Western blot in the livers of normal and transgenic mice of various ages. Changes in levels of superoxide dismutase (SOD), monoamine oxidase (MAO), malondialdehyde (MDA) as well as oxidative and anti-oxidative ability were measured.

RESULTSHistologically, more fatty degeneration and collagen deposition were found in the aging livers of transgenic mice than in those of the normal mice as their age of months increased. The mRNA and protein expressions of TIMP-1 were significantly high in the oldest animals. The histopathological changes, mRNA and protein expressions of TIMP-1 increased significantly in the liver of transgenic mice as compared with normal mice. The expression of MMP-2 and MMP-9 showed a minor change in the process of aging. Liver change and collagen deposition were not observed in young mice, but the activity of SOD decreased (P < 0.05), and the activity of MAO (P < 0.01) and the content of MDA increased in the liver of transgenic mice (P < 0.01).

CONCLUSIONSThe expression of TIMP-1 is significantly high in the liver of transgenic mouse in the process of aging, indicating that the oxidative level increases and the anti-oxidative ability decreases in the liver of transgenic mouse. TIMP-1 plays an important role in the process of liver aging.

Aging ; metabolism ; Animals ; Female ; Liver ; metabolism ; pathology ; Male ; Matrix Metalloproteinase 2 ; analysis ; genetics ; Matrix Metalloproteinase 9 ; analysis ; genetics ; Mice ; Mice, Transgenic ; Monoamine Oxidase ; analysis ; RNA, Messenger ; analysis ; Reactive Oxygen Species ; metabolism ; Superoxide Dismutase ; metabolism ; Tissue Inhibitor of Metalloproteinase-1 ; analysis ; genetics

OBJECTIVETo study the role of changes of matrix metalloproteinase-2, 9 (MMP-2, 9) activity in the development of dimethylnitrosamine (DMN)-induced liver fibrosis in rats.

METHODSThe rat liver fibrosis model was established by peritoneal injection of DMN (at a dose of 10 mg/kg, 3 times a week, for 4 weeks). The dynamic changes of liver fibrosis were observed at different time points (1d, 2d, 3d, 1 week, 2 weeks, 4 weeks, 6 weeks and 8 weeks). The MMP-2, 9 activity was measured by zymogram method. Liver ultrastructure was observed by electron microscope. The expressions of type IV collagen (CIV), laminin (LN), type I collagen (CI) and alpha-smooth muscle actin (alpha-SMA) were examined by immunohistochemistry. The tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) content was measured by Western blot method.

RESULTSThe MMP-2, 9 activity (gray value) significantly increased in the 2d and 3d DMN model rats (2d: normal/model group, MMP-2: 54.72+/-4.56/70.76+/-7.63; F = 16.27, P < 0.05; MMP-9: 25.72+/-4.29/51.76+/-15.33, F=13.38, P < 0.05). The positive staining area percentage of CIV in the sinusoidal walls decreased in the 2d, 3d and 1 weeks model rats (2d: normal/model group, 6.06+/-1.35/2.86+/-0.63, F=69.12, P < 0.05), but significantly increased in the 4w model rats (normal/model group, 6.06+/-1.35/8.04+/-1.50, F=14.42, P < 0.05). There was a remarkable negative correlation between the MMP-9 activity and expression of CIV in the sinusoidal walls (r = -0.729, P < 0.05). Positive expressions of LN and CI increased, and the strongest positive staining of them displayed in the 4w model rats. The formation of basement membrane was also observed in the 4 weeks model rats. Expression of TIMP-2 significantly increased in the late stage of fibrosis.

CONCLUSIONSThe increase of MMPs activity, especially MMP-9 which degrades the CIV normally distributed under the sinusoidal endothelium is the important factor in the formation of sinusoidal capillarization. The deposition and reconstitution of LN and new synthetic CIV, adding the deposition of CI constitute the high density basement membrane. The increase of TIMP-2 expression in the late stage of the fibrosis may be one of reasons why natural resolution of DMN-induced liver fibrosis is difficult.

Animals ; Collagen Type IV ; analysis ; Laminin ; analysis ; Liver ; chemistry ; ultrastructure ; Liver Cirrhosis, Experimental ; enzymology ; pathology ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Rats ; Rats, Wistar ; Tissue Inhibitor of Metalloproteinase-2 ; analysis

Asthma ; physiopathology ; Humans ; Inflammation ; etiology ; Matrix Metalloproteinase 9 ; analysis ; physiology ; Neutrophil Activation ; Neutrophils ; physiology ; Transforming Growth Factor beta ; physiology

BACKGROUNDThe aim of this study was to prospectively study the changes in neutrophil elastase (NE), fibroblast growth factor 9 (Fgf9), matrix metalloproteinase-9 (MMP-9), tissue inhibitor of metalloproteinase 1 (TIMP-1) in sputum induced during the early period after lung volume reduction surgery (LVRS).

METHODSFrom April to October 2005, ten consecutive patients with chronic obstructive pulmonary disease (COPD) underwent LVRS. Ten non-small cell lung cancer patients (stage II - IIIa) received lobectomy as a control group. The induced sputum was collected from both groups at six different times (two weeks before operation and postoperatively at 1, 2, 4, 6 and 10 days). The level of NE, Fgf9, MMP-9 and TIMP-1 were measured using enzyme-linked immunosorbent assay.

RESULTSThe pulmonary function (FEV(1)%) and arterial blood gases (PaO(2) and PaCO(2)) were significantly different between the groups. There were no significant differences in age, ejection fraction (EF), and operation duration, but hemoglobin in the LVRS group was statistically higher than in the controls. At certain times, there were significant differences in NE, MMP-9, TIMP-1 and MMP-9/TIMP-1 (P < 0.05) but not in Fgf9 between the two groups. The levels of NE and TIMP-1 were maximal at 2 days postoperatively and that of MMP-9 and MMP-9/TIMP-1 at 4 days postoperatively in the LVRS group. In the control group, maximal levels of NE and TIMP-1 occurred at 2 days postoperatively and that of MMP-9 and MMP-9/TIMP-1 at 1 day postoperatively. Ten days after surgery, all values of the control group were not significantly different from the baseline. In the LVRS group, the levels were significantly different from the pre-operative values (P < 0.05) apart from TIMP-1.

CONCLUSIONThe levels of NE, MMP-9, TIMP-1 and MMP-9/TIMP-1 of the LVRS group were different from those of the control group. The time course of these changes may be related to LVRS and the underlying process of COPD.

Female ; Fibroblast Growth Factor 9 ; analysis ; Humans ; Leukocyte Elastase ; analysis ; Lung Neoplasms ; surgery ; Male ; Matrix Metalloproteinase 9 ; analysis ; Middle Aged ; Pneumonectomy ; Prospective Studies ; Pulmonary Disease, Chronic Obstructive ; surgery ; Sputum ; chemistry ; Tissue Inhibitor of Metalloproteinase-1 ; analysis

OBJECTIVETo investigate the invasive biologic behavior of ameloblastoma (AB) and to analyze its correlative factors.

METHODSThe specimens of 43 cases of AB (primary AB 16 cases, recurrent AB 21 cases, malignant AB 6 cases) were examined immunohistochemically using the streptavidin-biotin method to determine the expression of E-cadherin (E-cad), matrix metalloproteinase (MMP)-2, MMP-9 and vascular endothelial growth factor (VEGF).

RESULTSThe cells in malignant AB scattered more, grew invasively, and the basal membrane ruptured or lost. The expression of E-cad in AB descended, MMP-2 and MMP-9 were strongly expressed in the epithelia cells of 28/41, 30/43 cases of AB, respectively. The positive rate and intensity of VEGF increased as AB recurred and transformed malignantly. The expression of E-cad, MMP-9 and VEGF were related to recurrence or malignant transformation of AB (r(s) = 0.309, 0.519, 0.381, P < 0.05).

CONCLUSIONAB is a high invasive tumor. The biological behavior of AB is related to lost or abnormal expression of E-cad, the high expression of MMP-2, MMP-9, and VEGF.

Adolescent ; Adult ; Aged ; Ameloblastoma ; chemistry ; pathology ; Cadherins ; analysis ; Child ; Female ; Humans ; Jaw Neoplasms ; chemistry ; pathology ; Male ; Matrix Metalloproteinase 2 ; analysis ; Matrix Metalloproteinase 9 ; analysis ; Middle Aged ; Neoplasm Invasiveness ; Vascular Endothelial Growth Factor A ; analysis