Aim To investigate estradiol inhibition of neointimal proliferation after rat carotid artery balloon injury. Methods Eight to ten-week-old SD rats (male,n=21,female,n=21) were divided into intact control(n=7),gonadectomy control(n=7) and estradiol (n=7, gonadectomy)groups in each sex. Left carotid artery was not injured with 2.0 F PTCA balloon until estradiol was injected for three days. Rats were killed 2 wk after injury. Neointimal areas and media area, ratios of intimal areas/media areas were measured with computer. Results Male neointimal areas and ratios of intimal areas /media areas in estradiol group were less than those in intact control group significantly(all P<0.01) and than those in gonadectomy control group (all P<0.05). Those in female,in estrdiol group were less than those in gonadectomy control group evidently (all P<0.01),and similar to those in intact control group (all P>0.05). Conclusions Estrdiol inhibits neointimal proliferation after the gonadectomy in rats undergoing carotid artery balloon injury.

Aim and Methods To investigate the effect of UCN-01(7-hydroxystaurosporine) on cell migration and invasion ability of DU-145, an invasive human prostate cancer cell line.Results It was found that UCN-01 at non-cytotoxic doses (100 nmol· L-1) significantly inhibited prostate cancer DU-145 cell invasion and migration behaviors.Moreover, this anti-invasion and migration activity of UCN-01 was associated with an up-regulation of cell adhesion molecule E-cadherin. Conclusion These results indicate for first time that UCN-01 inhibits the invasion and migration of human prostate cancer cells.Thus, clinical application of UCN-01 may contribute to the potential benefit for suppression of prostate cancer invasion and metastasis.

To evaluate the protective effect of anti-digoxin antiserum on hypoxic injury myocardium and its mechanism.Methods It was observed that different concentration anti-digoxin antiserum effect on endogenous digitalis-like factor and cell membrane ATPase activity in hypoxic myocardium model.Results The level of endogenous digitalis-like factor was remarkably higher,cell membrane ATPase activity were remarkably lower in hypoxic group than those of normal group;anti-digoxin antiserum can resume membrane ATPase activity.Conclusion Rise of endogenous digitalis-like factor was basic of molecular biology of myocardial damage during myocardial hypoxia.Anti-digoxin antiserum has lightened myocardial injury and has protective effect on hypoxic myocardium by against effect of endogenous digitalis-like factor.

To compare the anxiolytic effects of reduced metabolite of progesterone and benzodiazepine.Methods　The effects of allopregnanolone and diazepam on spontaneous locomotor activity and on exploration in the elevated plus-maze were studied in C57 mice 20 min after vehicle or drug intraperitoneal administration.Results Allopregnanolone (0.1 mg.kg-1,ip) elicited marked anxiolytic effects in terms of significantly reducing the latency to enter the open arm from (31.30±8.39)s to (8.80±6.00)s,(P<0.001),and significantly increasing both the number of open arm entries from 1.20 ± 0.42 to 4.80 ±1.75,(P<0.001) and the proportion of total time spent on the open arm from 7.13％ to 32.50％,(P<0.001).Meanwhile,the diazepam (0.25 mg·kg-1) produced a lower anxiolytic effect comparing to that of the allopregnanolone.Analysis of spontaneous locomotor activity showed while 0.5 mg·kg-1 of diazepam decreased the locomotor activity (P<0.01),neither 0.1 mg·kg-1 of allopregnanolone nor 0.25 mg·kg-1 of diazepam affect the locomotor activity score.Conclusion Together,these results provide evidence for differential behavioral actions of the neurosteroids and benzodiazepines.Since the allopregnanolone produce a selective anxiolytic effect without affecting the spontaneous locomotor activity,the allopregnanolone may be a better alternative for diazepam in treating anxiety.

AIM: To study the pharmacokinetics of phenolic acids from Mailuoning injection in rats. METHODS: SD rats were given a single i.v. administration dose of Mailuoning injection 10 mL/kg, plasma and urine were collected before and after injection. Phenolic acid components in plasma and urine were measured by LC/MS. Pharmacokinetic parameters were determined from the plasma concentration-time data and urinary excretion-time data with the DAS software package. RESULTS: After i.v. of Mailuoning injection, chlorogenic acid (CGA), 1, 5-dicaffeylquinic acid (1,5-DCQA), 3, 4-dicaffeylquinic acid (3,4-DCQA), 3, 5-dicaffeylquinic acid (3,5-DCQA) and caffeic acid (CA) were quickly excrectioned. The t1/2 of CGA, 1,5-DCQA, 3,4-DCQA, 3,5-DCQA and CA were 0.649, 0.334, 0.479, 0.486 and 0.330 h, respectively. AUC0-∞ were (22.522±2.716), (0.353±0.062), (3.620±1.246), (5.287±1.627) and (2.257±0.360) mg·L-1·h, respectively. After i.v. of Mailuoning injection, CGA, 1,5-DCQA, 3,4-DCQA, 3,5-DCQA and CA can all be detected in the urine. The amounts of CGA, 1,5-DCQA, 3,4-DCQA, 3,5-DCQA and CA excreted from urine during 0-24 h were (122.22±26.49)%, (3.30±1.26)%, (0.24±0.11)%, (1.93±0.77)% and (18.61±4.99)% of dose given in rats, respectively. CONCLUSION: After i.v. of Mailuoning injection, phenolic acids can be excreted quickly. Only a small quantity of 1,5-DCQA, 3,4-DCQA, 3,5-DCQA and CA were excreted from urine. 3,4-DCQA and 3,5-DCQA may be metabolized into CGA in the rat plasma.

AIM: To evaluate the protective effect of luteolin on endothelial dysfunction induced by tert-butyl hydro-peroxide (t-BOOH). METHODS: We observed the effect of luteolin on t-BOOH-induced contractions in the aorta rings with or without endot helium, which were incubated with luteolin (10-6 to 10-4 mol/L) for 30 min before determining the concentration-response to t- BOOH. Cultured endothelial cell line (ECV304) was pretreated with different concentrations of luteolin (10-6 to 10-4 mol/L) for 30 min and then exposed to 10 -5 mol/L t-BOOH for 24 hours. Cell morphology was observed , and cell viability was determined by MTT assay. Meanwhile, RT-PCR was used to measure the expression of eNOS and COX-1. RESULTS: Increasing concentrations of t-BOOH produced concentration-dependent cont ractions in aorta rings isolated from rats, luteolin effectively attenuated the contraction in a concentration-dependent manner, and the relaxation response was greater in intact endothelium segments. In MTT and RT-PCR assays, luteolin effectively reduced the cytotoxicity of t-BOOH to endothelium cell and increased the expression of nitric oxide synthase (eNOS) mRNA, which was greatly down-regulated by t-BOOH. CONCLUSION: Luteolin effectively protects the endothelium from the impairment of oxidative stress, and the protection could be related to its negative modulation towards t-BOOH-induced contractions in the aorta.

AIM: To evaluate the cytotoxicity of a novel anticholinergic drug penehyclidine hydrochloride (PHC) and its four optical isomers R-1, R-2, S-1, and S-2. METHODS: Two in vitro assays, MTT assay and neutral red uptake assay, were used to evaluate the cytotoxicity following PHC and its isomers exposure to HepG2 cells at different concentrations. RESULTS: PHC and its isomers induced decreases of viability of HepG2 cells in a concentration-dependent manner. Comparison of the cytotoxicity of the five anticholinergic agents with 50% inhibitory concentration (IC50) values indicated that the order of potency was PHC>R-2>R-1>S-2>S-1 for MTT assay, and R-2>PHC≈R-1>S-2>S-1 for neutral red uptake assay. CONCLUSION: With respect to the cytotoxicity of the four isomers on HepG2 cells, the R configuration was more potent than the S configuration, and R-2 was the most potent isomer whereas S-1 was the least potent isomer among the four optical isomers.

AIM: To evaluate the cardiac contractility and relaxation by Doppler tissue imaging (DTI) combined with myocardial contrast echocardiography (MCE) via injection of contrast media, Albunex. METHODS： Nineteen healthy mongrel dogs were conducted 60 min ligation of left anterior descending coronary artery (LAD), followed by reperfusion of 60, 120 and 180 min to establish an acute myocardial ischemic-reperfused canine model. (1) MCE was performed by bolus injection of Albunex at pre-reperfusion and at post-reperfusion. The perfused defect area defined by MCE at pre-reperfusion was regarded as risk area (RAMCE), while perfused defect area at post-reperfusion was regarded as no-reflow area (NRAMCE). When the ratio of NRAMCE to RAMCE exceeded 25%, myocardial reperfusion was considered incomplete, I.e., no-reflow group; If the ratio was <25%, myocardial reperfusion was considered adequate, I.e., reflow group. (2) Left ventricular ejection fraction (LVEF) and wall thickness ratio (△T%) of LV anterior wall were determined. (3)S-wave, e-wave and a-wave velocities at the LV anterior wall were determined by DTI. The e/a ratio was measured. RESULTS: The results of MCE showed 7 dogs in reflow group and 10 dogs in no-reflow group. (1) LVEF in reflow group gradually increased with time course after myocardial reperfusion, and in no-reflow group, however, LVEF increasingly declined with ongoing myocardial reperfusion. At the same reperfusion time point, LVEF of no-reflow group was significantly lower than that of reflow group. (2) △T% in reflow group improved gradually, and however, it can not come back to that of baseline at 180-min reperfusion. △T% in no-reflow group had no signal of recovery with progressive reperfusion. (3) S-wave, e-wave velocities measured by DTI significantly declined after ligation of LAD, and a-wave velocity increased, leading to decline of e/a. After myocardial reperfusion, s-wave, e-wave velocities and e/a in reflow group gradually increased at post-reperfusion, and a-wave velocity somewhat declined. In no-reflow group, on the other hand, s-wave, e-wave velocities and e/a progressively declined and a significant difference was present between reflow group and no-reflow group (P<0.05). CONCLUSION: Cardiac contractility and relaxation can not be recovered during myocardial microvascular impairment. This change may be further deteriorated with size enlargement of no-reflow area. DTI may provide a sensitive, reliable method for quantifying cardiac contractility and relaxation.

To establish an HPLC mehod for the analysis of pharmacokinetics of salvianolic acid B in rats. METHODS: The biological samples were extracted with acetic ether. The chromatographic conditions were as follows: Hypersil ODS column (200 mm×4.6 mm, 5μm) was used. The mobile phase was acetonitrile-water(with Ammoniom Acetate 0.25 mol/L) was set at 328 nm. RESULTS: Salvianolic acid B was injected intravenously at doses of 1.6, 3.2, 6.4 mg/kg. The terminal elimination half-life(t1/2) of α phase and β phase was (3.1±0.1) min and (31.5±3.2) min. The extents of excrement,urine and biliary excretion of salvianolic acid B were 1.43%±0.90%, 0.77%±1.01% and 8.82%±4.11%. The tissue concentration of salvianolic acid B was as followed in order: Cheart>Cliver>Clung>Cintestine>Ckidney>Cspleen>Cstomach. The plasma protein binding rate of salvianolic acid B in human plasma and in rat was similar(89.2%±1.8%,92.5%±1.5%). CONCLUSION: The method is accurate, stable and reliable, and can be used for the investigation of salvianolic acid B in pharmacokinetics research. Salvianolic acid B eliminates fast and it shows a high plasma protein binding rate, the mainly excretion way of salvianolic acid B is from biliary.

AIM: To illustrate the effects of drug transporters on the bile efflux of ibuprofen glucuronide(IBG), the difference of bile excretion and plasma concentration of ibuprofen(IB) and its glucuronides was studied in EHBR and normal SD rat(SDR). METHODS: After 20 mg/kg of IB enantiomers administrated intravenously, the bile and blood were collected from the rats and the concentration of IB and their glucuronide were measured by HPLC methods. RESULTS: The bile excretion of IBG was obviously (but no totally) suppressed in EHBR (1.7%±1.0%, 0.6%±0.9% of the dose respectively for S-IBG and R-IBG) compared with that in SDR (18.4%±4.0% and 3.0%±2.4% of the dose respectively for S-IBG and R-IBG), for both kinds of rats, there are more S-IBG excreted than that of R-IBG. As the result of reduction of IBG excreted in bile, the concentration of IBG was higher in blood in EHBRs. CONCLUSION: The results suggest that Mrp2 is the most important transporter for IBG, and other transporter(s) may participate in the process.