Aim To study the subtypes distribution of G-proteins and modulation by carbachol and pilocarpine on gene expression in endothelial cells.Methods The rat aorta endothelial cells were cultivated and incubated with carbachol and pilocarpine at the dose of 10~(-4) mol?L~(-1)for 6 h.Then the total RNA was extracted.The mRNA levels of G-protein ? subunits was measured by RT-PCR.Results Gq/11,Gs and Gi mRNA was detected in rat aorta endothelial cells,while G12/13 mRNA was not detected.Carbachol and pilocarpine treatment induced no changes in Gs,Gi and G11 mRNA.Gq mRNA was 72.7% up-regulated by carbachol and unchanged by pilocarpine.Conclusion In all G-protein ? subunits,only Gq mRNA was changed after activation of non-neuronal muscarinic receptor by carbachol.We can conclude that Gq-protein may play an important role in signal transduction of nonneuronal muscarinic receptor.

To examine the role and effect of nitric oxide synthase type II (NOS II) in cirrhotic rats, expression of NOS II mRNA was detected by real time RT-PCR. The enzymatic activity of nitric oxide synthase and the circulating levels of NO, systemic and portal hemodynamics and quantification of cirrhosis were measured. Chinese traditional medicine was used to treat cirrhotic rats and the effect of NO was evaluated. Double-blind method was used in experiment. Our results showed the concentration of NO and the enzymatic activity of NOS increased markedly at all stages of cirrhosis and iNOSmRNA was strongly expressed. Meanwhile, the portal-venous-pressure (PVP) and portal-venous-flow (PVF) were significantly increased. NO, NOS and iNOSmRNA were positively correlated to the degree of hepatic fibrosis. Tetrandrine significantly inhibited NO production and the expression of iNOSmRNA. Our results suggested that increased hepatic expression of NOS II is one of the important factors causing cirrhosis and portal hypertension. Tetrandrine can significantly ameliorate cirrhosis and portal hypertension.

The functions of vascular endothelial cells (VEC) are closely related with the etiology of cardiovascular diseases. The present paper reviews the bioactive substances secreted from endothelial cells and their regulatory effects on the tone of blood vessel wall, on the functions of platelet adhesion and aggregation, on the hemostatic and fibrinolytic systems.

Aim To investigate the effects of gene expression through activating endothelial target for acetylcholine (ETA).Methods Comparative studies were carried out to explore the distinct gene expression of carbachol and pilocarpine.Results Human coronary aortic endothelial cells were incubated with carbachol which activates ETA and muscarinic receptors and with pilocarpine which activates muscarinic receptors distinctly for 10 hours at the concentration of 100 ?mol?L -1.The endothelial gene expression was detected by BiostarH-14112S cDNA expression profiling microarrays containing 14 000 human unigenes. There were 801 differential genes totally (491 differentially expressed both by carbachol and by pilocarpine, 310 differentially expressed by carbachol exclusively). We found a significant increase in expression of 119 and a significant decrease in expression of 191 genes after treatment by carbachol preferentially. And these genes were not affected by pilocarpine. Further globally functional catagorization indicates that there are seven types of distinct expressed genes induced by carbachol selectively including membrane receptors and G proteins, ion channels, G protein coupled receptors, transcriptional factors, apoptosis-related genes, cell adhesive factors, thrombosis and atherosclerosis-related genes.Conclusion The differentially expressed genes evoked by carbachol exclusively are closely associated with ETA and its effectors.

There is another factor involving in endothelium dependent relaxation besides NO, which is independent of cGMP, EDHF. Its hyperpolarization mechanism may be related to the open of calcium dependent K + channels(K + Ca). NO, PGI 2,H 2O 2 and cytochrome P450 derived metabolite of arachidonic acid are all the candidates of EDHF. In the normal blood vessels, the influence of EDHF on endothelium dependent relaxation is little. But in some diseases, when the production and effect of NO is disturbed ,the mechanism of EDHF may play a specially important role.

AIM To examine antithrombotic effects of arecoline on the arterial thrombosis induced by carrageenin in mice through modulating the functions of endothelium and determine its mechanisms from hemostatic system, the platelet aggregative functions and the bioactive factors released by vascular endothelial cells. METHODS Kappa carrageenin was given ip in mice and mice were fed at the temperature of 20 to 21 degrees and at the humidity of 30 percent to 50 percent. RESULTS On the foregoing models of thrombosis, arecoline could antagonize the formation of thrombosis through activating the endothelial target for acetylcholine in a dose dependent manner and its antithrombotic potency was 250 to 500 times greater than aspirin; while under the same conditions, pilocarpine could not antagonize the formation of thrombosis. The levels of TT, PT, KPTT and MAR had no prominent changes compaired with control groups. The levels of t-PA became higher greatly than normal and the levels of PAI 1 became lower greatly than normal 2 hours after intravenous injection of arecoline in rats. Arecoline could decrease the higher plasma levels of thromboxane A2 and increase the lower plasma levels of prostacyclin in a dose dependent manner in the mice tail thrombosis induced by carrageenin. CONCLUSION The antithrombotic effects of arecoline are associated with activating the endothelial target for acetylcholine closely, but are not associated with muscarinic receptors,and not relevant to hemostatic systems or functions of platelet aggregation directly.

Objective: To investigate the effect of no-touch harvesting technique in reducing vein graft intimal hyperplasia. Methods: This longitudinal trial compared graft angiostenosis of two groups undergoing jugular vein to carotid artery interposition grafting in rabbit model. Conventional group: 12 rabbits had their veins stripped, distended, and stored in heparinized saline solution. No-touch group: 12 rabbits had veins removed with surrounding tissues, but were not distended, and stored in heparinized blood. The grafts were removed 4 weeks following grafting, and morphometry and immunohistochemistry assessment were performed. Results: The intimai thickness, degree of angiostenosis and proliferation index of vascular smooth muscle cells of no-touch group were significantly reduced (P<0.01) compared with those of the conventional group. The proliferating cell nuclear antigen positive-staining cells were significantly increased (P<0.01) in the conventional group compared with whose in the no-touch group. Conclusion: Harvesting the vein graft with no-touch harvesting technique could significantly reduce intimai hyperplasia of the vein graft.

Objective: To investigate the effect of no-touch harvesting technique in reducing vein graft intimal hyperplasia. Methods: This longitudinal trial compared graft angiostenosis of two groups undergoing jugular vein to carotid artery interposition grafting in rabbit model. Conventional group: 12 rabbits had their veins stripped, distended, and stored in heparinized saline solution. No-touch group: 12 rabbits had veins removed with surrounding tissues, but were not distended, and stored in heparinized blood. The grafts were removed 4 weeks following grafting, and morphometry and immunohistochemistry assessment were performed. Results: The intimai thickness, degree of angiostenosis and proliferation index of vascular smooth muscle cells of no-touch group were significantly reduced (P<0.01) compared with those of the conventional group. The proliferating cell nuclear antigen positive-staining cells were significantly increased (P<0.01) in the conventional group compared with whose in the no-touch group. Conclusion: Harvesting the vein graft with no-touch harvesting technique could significantly reduce intimai hyperplasia of the vein graft.

Aim To investigate the effects of newly synthesized 2, 3-dimethyl-2-butylamine derivatives on potassium channels in cortical neurons from rats.Method On cultured cortical neurons, the influences of those novel compounds on I A and I K currents were evaluated using whole-cell patch clamp technique. Result On cortical neurons, these compounds (5),(6),(7),(9) showed no effects on outward potassium currents, while the compounds (4),(8) exhibited inhibition of I K currents. But on a rterial smooth muscle cells(SMCs), all the six compounds had potassium channels opening activities. While the other three compounds had no effects on potassium channels of SMCs, the compound (3) decreased the I K currents evoked on neu rons.Conclusion The novel 2,3-dimethyl-2-butylamine derivati ves had the different modulation of potassium channels on cortical neurons as SM Cs.

Aim To establish a cell model heterologously expressing Kir6.x and SUR subunits of K_(ATP)channels and to study the selectivity of iptakalim on different subtypes of K_(ATP) channels.Methods Cell were transfected with the pcDNA vector containing the coding sequenced of Kir6.x and SUR using liposome and the pEGFP-N1 vector encoding for green fluorescent protein was added for easy identification of transfected cells.Using immunocytochemical technique,we detected the expression of Kir6.x and SUR protein in[FL(2K2]transfected cells.The electrophysiological experiment was performed after transfection 48-72 h.In whole cell configuration,the effects of K_(ATP) channel openers iptakalim,pinacidil and diazoxide on the clone currents in transfected HEK-293 cells and the antagonism of K_(ATP) channel inhibitor glibenclamide were evaluated.Results Immunocytochemical study identified the expression of Kir6.x and SUR protein in transfected cells.The electrophysiological experiment showed that in cells with recombinant expression of the Kir6.1/SUR2B channel complex,iptakalim(100 ?m),pinacidil(100 ?m)and diazoxide(200 ?m)significantly increased the clone current from(-88.0?30.8) pA to(-2042.6?127.3) pA,(-1431.9?142.4) pA and(-1104.7?228.9) pA,respectively,at-100 mV,and the actions were inhibited by glibenclamide(10 ?m).In cells expressed with Kir6.2/SUR2A channel,both iptakalim and pinacidil activated the currents,which was sensitive to glibenclamide.While diazoxide had no effects on the clone currents.In contrast,in cells with Kir6.2/SUR1channel,diazoxide increased the activity of recombinant K_(ATP) channels,while iptakalim and pinacidil had little effects.Conclusion From these observations,the effects of K_(ATP) channels openers with different chemical structures on the subtypes of K_(ATP) channels were diverse.Iptakalim showed selective action on Kir6.1/SUR2B and Kir6.2/SUR2A,but not Kir6.2/SUR1 K_(ATP) channels