Objective： To investigate the clinic values of combining test of serum matrix metalloproteinase 9 （MMP-9）, acetyl heparinase （Hpa） and Cathepsin L （CL） in diagnosis of ovarian cancer. Methods： Serum levels of...Objective： To investigate the clinic values of combining test of serum matrix metalloproteinase 9 （MMP-9）, acetyl heparinase （Hpa） and Cathepsin L （CL） in diagnosis of ovarian cancer. Methods： Serum levels of MMP-9, Hpa and CL were detected in a total of 418 cases, including 217 cases with ovarian malignant tumor, 100 cases with ovarian benign tumor and 101 healthy controls, by using enzyme-linked immunosorbent assay （ELISA）. Their correlation with clinicopathologic feature of ovarian malignant tumor was analyzed and their diagnosis performance was evaluated by receiver operating characteristic （ROC）. The combined diagnosis model was established by logistic regression analysis. Results： The serum levels of MMP-9, Hpa and CL were significantly higher in patients with ovarian malignant tumor than in benign tumor and healthy control, the serum levels of CL and Hpa were higher in epithelial cancer than in non-epithelial tumor, and MMP-9, Hpa and CL were elevated in low grade and advanced stage compared to high grade and early stage. The sensitivity for diagnosis of ovarian malignant tumor from high to low was CL, Hpa and MMP-9, and the specificity was MMP-9, CL and Hpa. The united diagnosis model was established and showed the sensitivity and specificity of combined detection were 84.6% and 82.1%, respectively, which were significantly higher than a single tumor marker. Conclusion： Serum MMP-9, Hpa and CL were correlated with ovarian malignant tumor and the combined detection of which may be valuable for clinical diagnosis of ovarian malignant tumor.展开更多
Heparinase Ⅲ(HepⅢ)is a 73-kDa polysaccharide lyase(PL)that degrades the heparan sulfate(HS)polysaccharides at sulfate-rare regions,which are important co-factors for a vast array of functional distinct proteins incl...Heparinase Ⅲ(HepⅢ)is a 73-kDa polysaccharide lyase(PL)that degrades the heparan sulfate(HS)polysaccharides at sulfate-rare regions,which are important co-factors for a vast array of functional distinct proteins including the well-characterized antithrombin and the FGF/FGFR signal transduction system.It functions in cleaving metazoan heparan sulfate(HS)and providing carbon,nitrogen and sulfate sources for host microorganisms.It has long been used to deduce the structure of HS and heparin motifs;however,the structure of its own is unknown.Here we report the crystal structure of the HepⅢ from Bacteroides thetaiotaomicron at a resolution of 1.6Å.The overall architecture of HepⅢ belongs to the(α/α)5 toroid subclass with an N-terminal toroid-like domain and a C-terminal β-sandwich domain.Analysis of this high-resolution structure allows us to identify a potential HS substrate binding site in a tunnel between the two domains.A tetrasaccharide substrate bound model suggests an elimination mechanism in the HS degradation.Asn260 and His464 neutralize the carboxylic group,whereas Tyr314 serves both as a general base in C-5 proton abstraction,and a general acid in a proton donation to reconstitute the terminal hydroxyl group,respectively.The structure of HepⅢ and the proposed reaction model provide a molecular basis for its potential practical utilization and the mechanism of its eliminative degradation for HS polysaccarides.展开更多
The novel heparinase-producing bacterial strain Sphingobacterium sp. was isolated and screened from soil. The optimum medium composition is （g/L）: Soytone 20, NaCl 1, K2HPO4 2.5, MgSO4 0.5, Heparin 2, Sucrose 15, pH...The novel heparinase-producing bacterial strain Sphingobacterium sp. was isolated and screened from soil. The optimum medium composition is （g/L）: Soytone 20, NaCl 1, K2HPO4 2.5, MgSO4 0.5, Heparin 2, Sucrose 15, pH 7.5. The optimum temperature for growth and enzyme production was 32℃. When cultured at a rotating shaker at 30℃ for 36 hours, 200r/min, 50mL medium in 500mL flask, the production of heparinase reached 4000U/L.展开更多
基金supported by a grant from the Provincial Research Project Funding of Guangxi,China (No. GSR 9817101)
文摘Objective： To investigate the clinic values of combining test of serum matrix metalloproteinase 9 （MMP-9）, acetyl heparinase （Hpa） and Cathepsin L （CL） in diagnosis of ovarian cancer. Methods： Serum levels of MMP-9, Hpa and CL were detected in a total of 418 cases, including 217 cases with ovarian malignant tumor, 100 cases with ovarian benign tumor and 101 healthy controls, by using enzyme-linked immunosorbent assay （ELISA）. Their correlation with clinicopathologic feature of ovarian malignant tumor was analyzed and their diagnosis performance was evaluated by receiver operating characteristic （ROC）. The combined diagnosis model was established by logistic regression analysis. Results： The serum levels of MMP-9, Hpa and CL were significantly higher in patients with ovarian malignant tumor than in benign tumor and healthy control, the serum levels of CL and Hpa were higher in epithelial cancer than in non-epithelial tumor, and MMP-9, Hpa and CL were elevated in low grade and advanced stage compared to high grade and early stage. The sensitivity for diagnosis of ovarian malignant tumor from high to low was CL, Hpa and MMP-9, and the specificity was MMP-9, CL and Hpa. The united diagnosis model was established and showed the sensitivity and specificity of combined detection were 84.6% and 82.1%, respectively, which were significantly higher than a single tumor marker. Conclusion： Serum MMP-9, Hpa and CL were correlated with ovarian malignant tumor and the combined detection of which may be valuable for clinical diagnosis of ovarian malignant tumor.
基金sup-ported in part by funds from Ministry of Science and Technology(No.2011CB910500)the National Natural Science Foundation of China(Grant No.31070661)+6 种基金the Natural Science Foundation of Zhejiang Province(No.R2100439)the Specialized Research Fund for the Doctoral Program of Higher Education(No.20110101110122)the Fundamental Research Funds for the Central Universities(SY)US Public Health Service grants(No.DK56338)(Texas Medical Center Digestive Diseases Center)Texas A&M Health Science Center Enhancement Grant(YL)CA05997P50 CA140388(WLM).
文摘Heparinase Ⅲ(HepⅢ)is a 73-kDa polysaccharide lyase(PL)that degrades the heparan sulfate(HS)polysaccharides at sulfate-rare regions,which are important co-factors for a vast array of functional distinct proteins including the well-characterized antithrombin and the FGF/FGFR signal transduction system.It functions in cleaving metazoan heparan sulfate(HS)and providing carbon,nitrogen and sulfate sources for host microorganisms.It has long been used to deduce the structure of HS and heparin motifs;however,the structure of its own is unknown.Here we report the crystal structure of the HepⅢ from Bacteroides thetaiotaomicron at a resolution of 1.6Å.The overall architecture of HepⅢ belongs to the(α/α)5 toroid subclass with an N-terminal toroid-like domain and a C-terminal β-sandwich domain.Analysis of this high-resolution structure allows us to identify a potential HS substrate binding site in a tunnel between the two domains.A tetrasaccharide substrate bound model suggests an elimination mechanism in the HS degradation.Asn260 and His464 neutralize the carboxylic group,whereas Tyr314 serves both as a general base in C-5 proton abstraction,and a general acid in a proton donation to reconstitute the terminal hydroxyl group,respectively.The structure of HepⅢ and the proposed reaction model provide a molecular basis for its potential practical utilization and the mechanism of its eliminative degradation for HS polysaccarides.
文摘The novel heparinase-producing bacterial strain Sphingobacterium sp. was isolated and screened from soil. The optimum medium composition is （g/L）: Soytone 20, NaCl 1, K2HPO4 2.5, MgSO4 0.5, Heparin 2, Sucrose 15, pH 7.5. The optimum temperature for growth and enzyme production was 32℃. When cultured at a rotating shaker at 30℃ for 36 hours, 200r/min, 50mL medium in 500mL flask, the production of heparinase reached 4000U/L.
