Biomedical Research ; Humans ; MicroRNAs ; Neoplasm Invasiveness ; genetics ; Neoplasm Metastasis ; genetics

Bleeding Time ; Child, Preschool ; Female ; Hemorrhage ; diagnosis ; Humans ; von Willebrand Diseases ; diagnosis

Objective:To investigate if chloride channel ClC-2 could be phosphorylated by mitogen activated protein kinase(MAPK) for further study of its regulation mechanism in proliferation and differentiation of cells.Methods:The coding sequence containing the cytosolic C-terminus of ClC-2 was amplified from pSPORT1/ClC-2 plasmid,including rabbit ClC-2 cDNA, by polymerase chain reaction(PCR),the fragment was cloned into pGEX-4T-1 plasmid for the construction of GST-tagged fusion protein expressing vector, pGEX-4T-1/ClC-2CT.After being identified by enzyme digestion and sequencing, the recombinant vector was transformed into a strain of E.coli BL21. The expression of GST-tagged fusion protein was induced with IPTG and purified with Gluthathion Sepharose 4B affinity chromatography. Then the phosphorylation of ClC-2 by MAPK was examined by using phosphorylation assays in vitro.Results:The construction of pGEX-4T-1/ClC-2CT recombinant vector was proved by enzyme digestion and sequencing. The purified fusion protein GST/ClC-2CT could be phosphorylated by MAPK, however the GST could not.Conclusion:Chloride channel ClC-2 can be phosphorylated by MAPK.

AIM: To evaluate the effect of tissue factor on intravascular migration of tumor cells.METHODS: Expression of tissue factor in tumor cells (HT1080) was analyzed by flow cytometry and immunofluorescence staining. An in vitro model was used to observe intravascular migration of tumor cells. RESULTS: High expression of tissue factor was observed in tumor cells (HT1080). The antibody for tissue factor inhibited intravascular migration of tumor cells. CONCLUSION: Tissue factor stimulated tumor metastasis through promoting intravascular migration of tumor cells.

Objective To evaluate the effect of hepatocyte growth factor(HGF) on transvascular metastasis of sarcoma cells.Methods The models of intravascular and extravascular migration of tumor cells in vitro were used to observe transvascular metastasis of sarcoma cells(HT1080).Results HT1080 migrated through basement membrane into blood vessels,and migrated through a gap between adjacent endothelial cells into extracellular matrix.The greater number of transmigrated HT1080 treated with HGF was observed(P

AIM: To evaluate the effect of endothelial myosin light chain kinase on extravasation migration of sarcoma cell. METHODS: An in vitro model of sarcoma cell transmigration across a monolayer of HUVEC cultured on collagen gel was used to observe extravasation migration of sarcoma cell and calculated the electrical resistance of HUVEC monolayer in extravasation migration of sarcoma cell. RESULTS: Sarcoma cell migrated through a gap between adjacent endothelial cells into collagen gel. The electrical resistance of a HUVEC monolayer reduced in extravasation migration of sarcoma cell.Endothelial myosin light chain kinase inhibitor(ML-7) inhibited extravasation migration of sarcoma cell and inhibited reduction of electrical resistance of a HUVEC monolayer in extravasation migration of sarcoma cell in a dose-dependent manner. CONCLUSION: Endothelial myosin light chain kinase regulates sarcoma cell transendothelial migration by phosphorylation of myosin light chain.

AIM: To evaluate the effect of endothelial Rho and Rho kinase in extravasation migration of sarcoma cell. METHODS: We used an in vitro model of sarcoma cell transmigration across a monolayer of HUVEC cultured on collagen gel to observe extravasation migration of sarcoma cells and calculated the electrical resistance of HUVEC monolayer in extravasation migration of sarcoma cells. RESULTS: Sarcoma cells migrated through endothelial cells into collagen gel, the electrical resistance of a HUVEC monolayer reduced in extravasation migration of sarcoma cells.Endothelial Rho inhibitor(C3 transferase) and Rho kinase inhibitor(Y-27632) inhibited extravasation migration of sarcoma cells and inhibited reduction of electrical resistance of a HUVEC monolayer in extravasation migration of sarcoma cells. CONCLUSION: Endothelial Rho and Rho kinase regulates sarcoma cell transendothelial migration through modification of endothelial cytoskeleton.

0.05). Phagocytic index of RPE cells was (28.7?1.9)% in the presence of 10 ?mol?L~ -1 DIDS,10 ?mol?L~ -1 DIDS significantly inhibited the nonspecific phagocytic process of human RPE cells(P

AIM:To investigate the protective effect of nerve growth factor combined with Ginkgo biloba extract on retinal ischemia-reperfusion (RIR) injury in rabbits with experimental high intraocular pressure.METHODS:Establishment of rabbit glaucoma ischemia reperfusion model.Twenty-four New Zealand white rabbits were randomly divided into three groups:nerve growth factor group, Ginkgo biloba extract group and combination group.Respectively, in the continuous administration of 1, 7, 14d.We observed the morphological changes of the tissues of the retina.The levels of superoxide dismutase(SOD), nitric oxide(NO) and malondialdehyde(MDA) in retinal tissue were measured.RESULTS:Respectively, first, in the continuous administration of 1, 7, 14d, the contents of MDA and NO in Ginkgo biloba extract group and nerve growth group were higher than that in combination group (P<0.05).Secondly, the SOD content of Ginkgo biloba extract group and nerve growth group were lower than that of combination group at each time point (P<0.05).At each time point, the number of HE staining of retinal ganglion cells (RGCs) showed that the loss of RGCs in the combination group was significantly lower than that in the other groups, and the ganglion cell count showed that the Ginkgo biloba extract group and the neuronal growth group were lower (P<0.05).CONCLUSION:Nerve growth factor combined with Ginkgo biloba extract has better protective effect on retinal ischemia-reperfusion injury.The mechanism may be related to the decrease of free radicals and increase the activity of SOD in retinal tissue.

Aged ; Epiglottis ; microbiology ; pathology ; Humans ; Larynx ; microbiology ; pathology ; Male ; Syphilis