Aim To investigate the in vitro vasorelax-ant effect of DL0805-0, a Rho kinase inhibitor, on iso-lated rat thoracic aorta and explore its underlying mechanism. Methods Tension was measured to eval-uate the vasorelaxant effect of DL0805-0 on rat endo-thelium-intact and endothelium-denuded thoracic aorta rings. Rho kinase inhibitor fasudil, nitric oxide syn-thase inhibitor Nω-nitro-L-arginine methyl ester ( L-NAME), guanylate cyclase inhibitor methylene blue, cyclooxygenase inhibitor indomethacin, calcium-activa-ted potassium channel blocker tetraethyl ammonium ( TEA ) , ATP-sensitive potassium channel blocker glibenclamide and voltage-dependent potassium chan-nel blocker 4-aminopyridine ( 4-AP ) were used to il-lustrate the mechanisms of vasorelaxant effect of DL0805-0 . Results DL0805-0 exerted vasorelaxation in a dose-dependent manner in KCl (60 mmol·L-1 ) or NE ( 0. 1 μmol · L-1 ) -induced contraction. DL0805-0-induced vasorelaxation was significantly re-duced by L-NAME. However, methylene blue and in-domethacin did not significantly affect vasorelaxation of DL0805-0. In endothelium-denuded rings, TEA re-markably attenuated the vasorelaxant effect of DL0805-0 , while glibenclamide and 4-AP did not affect vasore laxation of DL0805-0 significantly. DL0805-0 also re-duced NE-induced transient contraction and inhibited contraction induced by increasing extracellular calci-um. Conclusion These results suggest that DL0805-0 induces vasorelaxation through an endothelium-depend-ent pathway. The opening of calcium-activated K+channels and blocking of Ca2+ channels in vascular smooth muscle cells may be one of the mechanisms of DL0805-0-induced vasorelaxation.

Aim ToestablishthemethodofHighper-formance liquid chromatography ( HPLC ) for detecting plasma concentration of indazole compound DL0805-1 , a Rho kinase inhibitor, and to investigate its pharma-cokinetics in rats with intravenous injection. Methods ThedetectingsystemwasAgilent1200-DAD;chro-matographic column was Agilent TC-C18 ( 4. 6 mm × 250 mm, 5 μm); the ultraviolet detection wavelength was 235 nm; the column temperature was 35 ℃; the flow rate was 1 ml·min-1;the mobile phase was ace-tonitrile-0. 05% H3 PO4 gradient elute. Rat blood sam-ples were collected at different intervals after intrave-nous injection of a single dose of DL0805-1 , and the concentration of DL0805-1 in rat plasma were deter-mined by HPLC method for estimating pharmacokinetic parameters.Results Afterintravenousinjectionof DL0805-1 in rats, prototype and its metabolite were detected in plasma. T1/2 of DL0805-1=(2. 34 ± 1. 42) h, Cmax=(3. 51 ± 0. 44) mg·L-1, T1/2 of metabolite of DL0805-1 = ( 1. 27 ± 0. 45 ) h, Cmax = ( 3. 55 ± 0.22)mg·L-1.Conclusion Theseresultssuggest that DL0805-1 may be metabolized into another sub-stance in vivo and play biological functions. The meth-od is sensitive, simple, and accurate, and can be used for the determination of DL0805-1 in rat plasma and pharmacokinetic studies.

Aim To evaluate the vasorelaxant effect of two new chemical entities, J35242 and J35243, on iso-lated rat thoracic aorta rings as Rho-kinase inhibitors, and further to explore the underlying mechanisms of these two compounds. Methods Isolated rat thoracic aorta rings pre-contracted by KCl or norepinephrine ( NE) were used to evaluate the vasodilatory effect of J35242 and J35243 . Through the interventions of sev-eral tool drugs, the mechanisms of compounds concern-ing endothelium, K+ channels and Ca2+ were studied. Results J35242 and J35243 showed potent relaxant effect on both KCl and NE pre-contracted vessels, and exhibited partial endothelium dependency. L-NAME and Methylene Blue( MB) could influence the relaxant effect of these compounds. Meanwhile, the compounds could inhibit intracellular Ca2+ release and extracellu-lar Ca2+ influx, which indicated that the compounds might block the calcium channels to relax the vessels. In addition, the two compounds probably did not dilate the aorta rings through opening potassium channels. Conclusions J35242 and J35243 have vasorelaxant effects on vessels in vitro and the potency of J35242 is stronger than that of J35243 . The underlying mecha-nisms might be endothelium-dependent. Also the com-pounds might block Ca2+ channels, lowering intracel-lular Ca2+ concentration to relax the vessels.

Objective: To detect the expression level of astrocyte elevated gene-1 (AEG -1) in human bladder cancer cell lines, and to investigate the effects of down-regulation of AEG-1 expression on proliferation and invasion of bladder cancer T24 cells. Methods: The expression level of AEG-1 protein in normal bladder urinary epithelial cell line SV-HUC-1 and bladder cancer cell lines RT4, J82, 5637 and T24 was detected by Western blotting and immuno?uorescence staining. The recombinant virus with specific shRNA targeting AEG -1 gene was infected into bladder cancer T24 cells with high-expression of endogenous AEG-1. The stable infected clones were screened by puromycin, and the silencing efficiency of AEG -1 gene was confirmed by Western blotting. Then the effects of AEG -1 gene-silencing on the cell proliferation, migration and invasion were examined by CCK-8 method, wound healing assay and Transwell chamber assay, respectively. Results: The expression levels of AEG-1 in 4 bladder cancer cell lines were significantly higher than that in normal bladder urinary epithelial cell line (all P < 0.05). After infection AEG-1 shRNA with the recombinant virus carrying AEG-1 shRNA, the expression level of AEG-1 protein was effectively down-regulated in bladder cancer T24 cells (P < 0.05). As compared with the negative shRNA transfection group, the proliferation of T24 cells after AEG -1 genesilencing was obviously inhibited (P < 0.05), and the migration and invasion abilities of T24 cells after AEG -1 gene-silencing were significantly decreased (both P < 0.05). Conclusion: Silencing AEG -1 gene expression can inhibit the proliferation, migration and invasion of bladder cancer cells, which suggests that AEG-1 maybe paly a role in promoting the malignant biological behavior of bladder cancer cells.

Objective To determine the value of acute phase protein in the early diagnosis of recurrence after curative gastric cancer surgery. Methods Acute phase serum protein level was measured before and after the gastric cancer surgery in 120 patients,and was compared with that of the control group. At 1, 3, 6, 9, 12 months after the operation in 87 patients undergoing curative gastric cancer surgery, the levels of acute phase proteins were measured. They were followed up for at least 12 months or until death. Results The levels of serum C reactive protein(CRP), ?1 antitrypsin (?1 AT) and ? acid glycoprotein (? AG) in gastric cancer group were significantly higher than those in the control group (P0.05). In patients who underwent curative gastric cancer surgery in a certain period after the operation, the serum levels of CRP, ?1 AT and ? AG were significantly lower than those before the surgery(P0.05).There were significant difference in CRP,?1 AT and ? AG between the recurrence groups and nonrecurrence group before and after the operation (P