OBJECTIVE: To investigate the effect of form-deprivation on level of gelatinase in the posterior sclera in chicks. METHODS: Fifty 1-day-old chicks were monocularly deprived to establish the animal model of form-deprivation myopia (FDM). According to the duration of form-deprivation the experimental chicks were divided randomly and equivalently into 5 groups, which were deprived for 3, 7, 14, 21 and 30 days respectively. Meanwhile the other eyes of the deprived chicks were used as self-control groups and chicks of the same days were chosen randomly as the normal control groups for each FDM group. At each form-deprivation point the changes of degree of diopters and axial length of chicks in each group were recorded. The levels of gelatinase in posterior sclera of the experimental eyes were measured by gelatin enzymography. RESULTS: Compared with the normal and self-control groups, the levels of MMP-2 activity in FDM groups were much higher (P <0.01). With the increase of the time of monocular deprivation these changes became more significant and reached the top after 14 days' deprivation with an inter-group statistical difference (P <0.01). The dynamic changes of MMP-2 activity were the same as those of axial length and degree of diopters in each experimental groups. There was positive correlation between the MMP-2 activity and axial length (r = 0.989, P < 0.01). But there was a negative correlation between the MMP-2 activity and refractive degree. CONCLUSION Increase of MMP-2 activity in the posterior sclera of chicks would be a direct key factor to trigger sclera ECM remodeling process in chick FDM.
Animals ; Chickens ; Gelatinases ; metabolism ; Matrix Metalloproteinase 2 ; metabolism ; Myopia ; enzymology ; etiology ; Sclera ; enzymology

OBJECTIVETo investigate the time course of changes in the expression of fibroblast activation protein (FAP) during healing of skin scald burns in rats.

METHODSAdult Wistar rats were randomized into two equal groups (n=42) and subject to superficial second degree and deep second degree scald burns on the dorsal skin groups, with 6 normal rats serving as the control group. At 6 h, 12 h, and 1, 3, 7, 14, and 21 days after burns, 6 rats from each group were sacrificed to detect FAP expression by immunohistochemistry and Western blotting.

RESULTSFAP was expressed on the cell membrane and in the cytoplasm of the fibroblasts, especially those around the neovessels. In both burn groups, FAP expression increased significantly at 6 h after burns. In superficial burn group, FAP expression was comparable between 6 and 12 h and between 1 and 3 days (P>0.05), but showed significant differences between the other time points (P<0.05). In deep burn group, FAP expression was comparable between 12 h, 1 day and 3 days (P>0.05) but differed significantly between the other time points (P<0.05). In both burn groups, FAP expression reached the peak level at 7 days followed by a gradual declination. At 21 days after the burns, FAP maintained a significantly higher expression level than the control level (P<0.05).

CONCLUSIONThe time course of the changes of FAP expression following scald burns suggests an important role of FAP in the healing process of scald burns.

Animals ; Burns ; metabolism ; rehabilitation ; Face ; Gelatinases ; metabolism ; Membrane Proteins ; metabolism ; Rats ; Rats, Wistar ; Serine Endopeptidases ; metabolism ; Skin ; metabolism ; Wound Healing

OBJECTIVETo investigate the relationship between the expression of the quorum-sensing related genes during Enterococcus faecalis(Ef) biofilm formation.

METHODSEf biofilms model was established in vitro and film formation process was observed by confocal laser scanning microscope at 6, 12, 24 and 48 hours respectively.Quantification of biofilms was achieved by staining with crystal violet.Real-time fluorescence quantitative PCR method was used to detect the expression of fsrB, gelE and sprE genes in the process of Ef biofilm formation.

RESULTSA lot of live and dead bacteria unevenly distributed in Ef biofilm. The quantity of biofilms increased with time within 24 hours and was 0 h:0.00 ± 0.00, 6 h:1.09 ± 0.13, 12 h:2.10 ± 0.79, 24 h:3.30 ± 0.13, which was significantly different among the 4 time period(P < 0.05). The quantity of biofilm at 48 h(3.51 ± 0.01) increased slightly compared with 24 h(3.30 ± 0.13) , but did not show significant difference.Quantitative real-time PCR showed that the expression of quorum-sensing related fsrB increased with time within 24 hours and was 0 h:9.98 ± 0.46, 6 h:23.45 ± 1.13, 12 h:47.30 ± 2.49, 24 h: 331.30 ± 2.18, which was significantly different among the 4 time period(P < 0.05). The expression of gelE was 0 h: 6.54 ± 0.73, 6 h: 14.26 ± 1.24, 12 h: 37.47 ± 2.35, 24 h:264.80 ± 5.10(P < 0.05). The expression of sprE was 0 h: 7.72 ± 0.74, 6 h: 21.15 ± 0.96, 12 h:49.87 ± 3.18, 24 h:441.89 ± 7.74, which was significantly different among the 4 time period(P < 0.05).

CONCLUSIONSThe fsrB, gelE and sprE genes are closely related to the biofilm formation in Ef.

Bacterial Proteins ; metabolism ; Biofilms ; growth & development ; Enterococcus faecalis ; genetics ; metabolism ; physiology ; Gelatinases ; metabolism ; Gene Expression Regulation, Bacterial ; Genes, Bacterial ; Quorum Sensing ; Serine Proteases ; metabolism

OBJECTIVETo detect the expression of fibroblast activation protein(FAP) in colorectal cancer tissue, and to investigate the association between expression of FAP with pathological parameters.

METHODSFifty-five cancer tissues and 50 normal colorectal samples were examined using immunohistochemistry with anti-FAP polyclonal antibody. The distribution of positive cells in different tissues, and associations of positive cell number with tumor staging, lymph node metastasis and tumor invasion were investigated to evaluate the effects of FAP on pathological progress in colorectal cancer.

RESULTSNo FAP expression was observed in 50 normal colorectal tissue samples. FAP positive cells were seen in carcinoma associated fibroblasts(CAFs), and in few colorectal cancer cells. The numbers of FAP positive cells in tissue samples of TNM III(-IIII((40.1±15.9) was significantly greater than that of TNMI(-II( (18.3±7.7)(P<0.01). Furthermore, the number of FAP positive cells in tissue samples with lymph node metastasis (44.4±13.3) was also significantly higher than those without lymph node metastasis (18.5±8.1)(P<0.01). Significant positive correlations were found between the number of FAP-positive cells with the tumor TNM staging and lymph node metastasis(r=0.544 and r=0.793, respectively)(P<0.01). The number of FAP-positive cells was 25.2±8.9 in T2, 32.41±19.30 in T3, and 29.2±16.5 in T4. The association between number of positive cells and depth of invasion was not statistically significant(P>0.05).

CONCLUSIONSThe FAP mainly expresses in CAFs locating in colorectal cancer tissues. The number of FAP positive cells is positively correlated with TNM staging of colorectal cancer and lymph node metastasis.

Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Colorectal Neoplasms ; metabolism ; pathology ; Female ; Gelatinases ; metabolism ; Humans ; Lymphatic Metastasis ; Male ; Membrane Proteins ; metabolism ; Middle Aged ; Serine Endopeptidases ; metabolism

OBJECTIVETo evaluate the immunophenotype conversion of fibroblasts and its clinical significance in the process of breast tumor stromal remodeling.

METHODSCD34, FAP-α, p63 and a-SMA were detected by immunohistochemistry in 273 breast biopsies, including 60 normal breast tissues, 46 atypical ductal hyperplasia (ADH), 60 ductal carcinoma in situ (DCIS), 47 DCIS microinvasive carcinoma (DCIS-MI) and 60 invasive ductal carcinoma (IDC).

RESULTSThe positive expression rates of CD34, FAP-α and α-SMA in the stromal fibroblasts of normal breast tissues were 93.3%, 6.7% and 18.3%, respectively. Those in the stromal fibroblasts of ADH tissues were 95.7%, 4.3% and 10.9%, respectively. Those in the stromal fibroblasts of DCIS tissues were 95.0%, 8.3% and 15.0%, respectively. Those in the IDC tissues were 35.0%, 85.0% and 93.3%, respectively. The expressions of CD34, α-SMA and FAP-α in the stromal fibroblasts of normal, ASH and DCIS breast tissues did not show significant differences (χ(2) = 1.142, P = 0.896). The main immunophenotype of stromal fibroblasts in the tumor-host interface at the invasive front of ADH and DCIS lesions was CD34(+)α-SMA(+)FAP-α(+). There were statistically significant differences in the expression of CD34, α-SMA and FAP-α between IDC and ADH, DCIS and normal breast tissues (χ(2) = 8.351, P < 0.001). The immunophenotype of stromal fibroblasts in the IDC and DCIS-MI breast tissues was CD34(-) α-SMA(+) FAP-α(+).

CONCLUSIONSImmunophenotype conversion from CD34(+) α-SMA(-) FAP-α(-) to CD34(-) α-SMA(+)FAP-α(+) may be a sensitive indicator to judge whether DCIS has microinvasion. Detection of the immunophenotype conversion of stromal fibroblasts may be helpful to determine the presence of microinvasion, and to improve the diagnostic accuracy rate of DCIS.

Breast ; Breast Neoplasms ; immunology ; pathology ; Carcinoma in Situ ; Carcinoma, Ductal, Breast ; Carcinoma, Intraductal, Noninfiltrating ; Fibroblasts ; immunology ; Gelatinases ; metabolism ; Humans ; Hyperplasia ; Immunohistochemistry ; Immunophenotyping ; Membrane Proteins ; metabolism ; Serine Endopeptidases ; metabolism

Carcinoma, Squamous Cell ; metabolism ; pathology ; Chemokine CCL2 ; metabolism ; Fibroblast Growth Factor 7 ; metabolism ; Fibroblasts ; metabolism ; pathology ; Gelatinases ; metabolism ; Humans ; Membrane Proteins ; metabolism ; Mouth Neoplasms ; metabolism ; pathology ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Serine Endopeptidases ; metabolism

OBJECTIVETo study the effect of glycyrrhetinic acid (GA) on the invasive capacity of leukemic cells and the activity of intracellular gelatinase.

METHODSThe effect of GA, in different concentrations, on the proliferation of cultured K562 and HL-60 leukemic cells in vitro was determined by MTT assay; that on cell invasive capacity was tested by Transwell cubicle matrigel invasion assay; and that on the activity of gelatinase in cells was detected by gelatin zymography.

RESULTSGA showed significant inhibitory effect on the proliferation of K562 and HL-60 leukemic cells; it inhibited the invasive capacity of cells in concentration-dependent manner; and significantly down-regulated the activity of gelatinase A and B in cells.

CONCLUSIONSGA can inhibit invasive capacity of K562 and HL-60 leukemia cells by way of suppressing the activity of gelatinase A and B. This study provides an experimental evidence for preventing extra-medullary infiltration of leukemic cells.

Cell Proliferation ; drug effects ; Down-Regulation ; Gelatinases ; metabolism ; Glycyrrhetinic Acid ; pharmacology ; HL-60 Cells ; Humans ; K562 Cells ; Leukemia ; drug therapy ; enzymology ; pathology ; physiopathology ; Neoplasm Invasiveness

AIMTo investigate the relationship between the expression of MMP-2, MMP-9, TIMP-1 and TIMP-2 and ECM accumulation in rat left ventricle in a mechanical unloaded heart model.

METHODS12-week-old male Lewis rats were subjected to abdominal heterotopic heart transplantation to achieve pressure and volume unloading(mechanical unloading). Age and sex matched in situ heart of Lewis rats were used as control. Collagen volume fraction(CVF) was analyzed by picrosiris-red staining plus polarized microscopy. MMP-2 and -9 gelatinolytic activity were measured by gelatin-zymography. mRNA level of MMP-2, MMP-9, TIMP-1 and TIMP-2 were measured by real-time quantitative PCR. TIMP-1 and TIMP-2 protein level were measured by immunoblotting.

RESULTSMyocardial cross-sectional area of transplanted heart was significantly reduced, and accompanied by excessive ECM deposition (CVF 5.22% +/- 1.6% vs. 2.21% +/- 0.9%, P < 0.05) compared to in situ heart. MMP-2 and MMP-9 activity were significantly increased, as well as mRNA level of MMP-2, MMP-9, TIMP-1 and TIMP-2 compared to in situ heart. TIMP-1 and TIMP-2 protein level in mechanically unloaded heart were significantly upregulated compared to in situ heart, especially for TIMP-1.

CONCLUSIONMechanical unloading of left ventricle may lead to excessive ECM deposition, accompanied by imbalance between MMPs and TIMPs system, especially the upregulation of TIMPs.

Animals ; Extracellular Matrix ; metabolism ; Gelatinases ; metabolism ; Heart Transplantation ; physiology ; Heart-Assist Devices ; Male ; Matrix Metalloproteinases ; metabolism ; RNA, Messenger ; metabolism ; Rats ; Rats, Inbred Lew ; Tissue Inhibitor of Metalloproteinases ; metabolism ; Transplantation, Heterotopic ; physiology ; Ventricular Dysfunction, Left ; metabolism

Animals ; Collagenases ; metabolism ; Dental Bonding ; Dental Caries ; enzymology ; Dentin ; enzymology ; pathology ; Gelatinases ; metabolism ; Humans ; Matrix Metalloproteinase 14 ; metabolism ; Matrix Metalloproteinase 20 ; metabolism ; Matrix Metalloproteinase 3 ; metabolism ; Matrix Metalloproteinases ; metabolism ; Sclerosis

UNLABELLED: OBJECTIVE To investigate the time-dependent expression of fibroblast activation protein (FAP) and alpha-smooth muscle actin(alpha-SMA) during the incised wound healing of the skin in mice. METHODS: The expression of FAP and alpha-SMA in incised wound of mice skin was detected by immunohistochemistry and Western blot. RESULTS: By immunohistochemistry, the expression of FAP and alpha-SMA in the normal skin and the skin 1 h after injury maintained at a very low level, but the positive cells expressing FAP and alpha-SMA started to elevate 6 h after injury and reached its peak on 5 d for FAP and on 3 d for alpha-SMA, then gradually decreased to the normal level on 14 d. The expression of FAP and alpha-SMA was observed throughout the wound healing stages 1 d after injuries by Western blot as well with a peak expression occurring on 5 d for FAP and on 3 d for alpha-SMA after injury. CONCLUSION FAP may be a potentially useful marker for wound age determination and alpha-SMA may be used as an effective indicator for the mid- and late stage incised wound of mice skin. The combination use of FAP and alpha-SMA may be potentially effective indicators for wound age determination.
Actins/metabolism* ; Animals ; Blotting, Western ; Disease Models, Animal ; Endopeptidases ; Female ; Fibroblasts/metabolism* ; Forensic Pathology ; Gelatinases/metabolism* ; Immunohistochemistry ; Male ; Membrane Proteins/metabolism* ; Mice ; Random Allocation ; Serine Endopeptidases/metabolism* ; Skin/metabolism* ; Time Factors ; Wound Healing ; Wounds and Injuries/physiopathology*