Objective To investigate the changes in discharge rates of pain-sensitive neurons (PSN) in thalamic parafaseicular nucleus ( Pf) following coronary artery occlusion (CAO) and the effects of urapidil, a partial 5-HT agonist. Methods One-hundred male SD rats weighing 260-300 g were operated upon under urethane anesthesia and local infiltration of the skin incision. The animals were tracheotomized and mechanically ventilated. A hole was drilled in the skull until the brain was exposed. A single-barrel glass electrode was inserted, aiming at the PSN, the discharges of which were filtered, amplified and recorded. Chest was opened and heart was exposed. A tie was placed around the anterior descending branch of left coronary artery which can be occluded whenever needed. The study was divided into 3 groups : group I CAO alone; group II CAO + urapidil and group III CAO + urapidil + methysergide ( a potent serotonin antagonist). Urapidil 0.21 mg.kg-1 was given intravenously 15 min after CAO. Methysergide 0.1 mg.kg-1 was given iv 20 min after urapidil. Results Discharges of PSN were recorded in 45 animals out of the 100, and the recordings were complete for investigation in 31 animals. CAO evoked significant increase in the discharge frequency of PSN in 18/31 animals. After intravenous urapidil the increased frequency of nociceptive discharges was inhibited; however intravenous methysergide could partially antagonize the antinociceptive effect of urapidil. Conclusion The results indicate that (1 )the nociceptive response could be induced by CAO in rats; (2) Pf nucleus of thafamus is involved in the myocardial ischemia-induced nociceptive response of central nervous system; (3) serotonin plays a critical role in the modulation of the nociceptive signal of acute myocardial ischemia.

Objective:To evaluate the application of puncturing the umbilical cord under the ultrasound guidance in prenatal diagnosis,clinical high risk factors of fetal chromosomal abnormalities were discussed.Methods:283 pregnant women who had the percutaneous umbilical cord blood sampling were recruited and the karyotyping was done.The detection rate of chromosomal abnormalities and correlate factors in different subgroups of prenatal diagnosis were analyzed.Results:The success rate of puncture was 100%.Major complications were bleeing at the puncture site(33 cases,11.67%)and fetal heart brachycadia(5 cases,1.77%).25 chromosomal abnormalities were found and the detection rate was 8.83%.There was significant correlation between the couple's chromosomal abnormalities and the fetal's(r=22.348,P=0.000).Conclusions:The ultrasound guided percutaneous umbilical cord blood sampling proves to be valid in the prenatal diagnosis,especially when couples have chromosomal abnormalities.

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

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 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 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.

10.3969/j.issn.1000-8179.2013.12.004

Objective To analyze the influence of dexamethasone in the hypotonic-induced volume-sensitive chloride currents in human trabecular meshwork cells,and to investigate the possible mechanism of volume-sensitive chloride channels(VACC)in the glucocorticoid-induced glaucoma(GIG)cases.Methods The human trabecular meshwork cells were seeded in 35 mm diameter plastic petri dishes,so that they could grow in monolayer.The cultured cells were divided into normal cell culture medium group and dexamethasone 1 d,3 d,7 d groups.The chlorine current density values of the cells in four groups were recorded respectively by the whole-cell patch-clamp technique. The differences among groups were compared.Results In normal group,after hypotonic stimulation,under+100 and-100 mV voltage clamp,the outward and inward current density values of the trabecular cells were (19.94±0.87) and (-6.53±0.41)pA/pF.In dexamethasone 1 d,3 d,and 7 d groups,under the same condition,the outward and inward current density values of the trabecular cells were (19.39 ± 1.40)and (-6.42 ± 0.28)pA/pF, (17.97±2.35)and (-5.82±0.94)pA/pF,(17.16±1.16)and (-5.65±0.43)pA/pF.The trabecular cells cultured with dexamethasone for 1 d had lower outward and inward current density values under hypotonic stimulation compared with normal group,but there was no significant difference (P>0.05).The trabecular cells cultured with dexamethasone for 3 d and 7 d,when compared with normal group,had significantly lower outward and inward current density values under hypotonic stimulation (P<0.05 ). Conclusion Dexamethasone could reduce the volume-sensitive chloride current in trabecular meshwork cells,which would affect trabecular meshwork cell volume adjustment.This would possibly cause the increase of the aqueous humor outflow resistance among GIG cases.

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 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.