Objective: To establish a UPLC-MS/MS analysis method for determination of baicalin, geniposide, chlorogenic acid, cholic acid and hyodeoxycholic acid in Qingkailing (lyophilized) for injection in rat plasma, and to investigate the pharmacokinetic behavior of this preparation in normal and cerebral ischemic rats. Method: Rats were randomly divided into normal group and cerebral ischemia model group. The rat model of cerebral ischemia was established by suture embolization. The rats were given by intraperitoneal injection, and normal saline was used as the solvent. Blood samples were taken at the corresponding time points. After treatment, UPLC-MS/MS was used to determine the blood concentration of five components. The main detection conditions were mobile phase of 0.1%formic acid aqueous solution-acetonitrile for gradient elution (0-0.25 min, 90%A; 0.25-1 min, 90%-75%A; 1-2 min, 75%-50%A; 2-2.6 min, 50%-45%A; 2.6-2.65 min, 45%-90%A; 2.65-4.0 min, 90%A), the flow rate of 0.4 mL·min^-1, the column temperature at 40℃, electrospray ionization under negative ion mode. The pharmacokinetic parameters were fitted and the bioavailability was calculated, the differences of treatment process of five components from Qingkailing (lyophilized) for injection in normal and cerebral ischemic rats were analyzed. Result: Compared with the normal group, the area under the curve (AUC_0-t) of geniposide in rats from cerebral ischemia model group decreased significantly after intraperitoneal injection of Qingkailing (lyophilized) for injection (P<0.05), and the time to peak (T_max) of chlorogenic acid in rats from cerebral ischemia model group was significantly earlier than that in the normal group (P<0.01). Pharmacokinetic parameters of baicalin, cholic acid and hyodeoxycholic acid had no significant difference between these 2 groups. Conclusion: Qingkailing (lyophilized) for injection has a certain difference in the treatment process between normal and cerebral ischemic rats, which has certain guiding significance for the clinical treatment of cerebral ischemic diseases with this preparation.

Objective:To discuss effect of nicorandil on unstable angina patients With persistent Weakly positive for troponin I (TnI).Methods:A total of 111 unstable angina patients With persistent Weakly positive for TnI Were randomly divided into control group (received routine treatment,55 cases) and intervention group (received nic-orandil 5mg,3 times/d based on routine treatment,56 cases).The relief of chest pain in one Week,the recurrent hospitalization for chest pain aggravation in 3 months and the cardiac mortality rate in one year betWeen tWo groups Were observed in tWo groups. Results:Compared With control group,the relief of angina pectoris in one Week (63.6% vs. 91.1%,χ2=11.97,P=0.0005)significantly increased,re-hospitalization for chest pain aggravation in three months (56.4% vs.19.6%,χ2=15.91,P=0.0001)significantly decreased in intervention group;but cardiac mortality rate during one year betWeen tWo groups Was no significant difference (5.5% vs. 8.9%,χ2=0.50,P=0.4792).Conclusion:Nicorandil can significantly reduce the unstable angina and re-hospitalization for chest pain aggravation in patients With persistent Weakly positive for TnI,but there Was no significant difference in reducing mortality Within one year betWeen tWo groups.

Objective To investigate the protective effect and mechanism of epigallocatechin-3-gallate against myocardial ischemia and reperfusion injury in rats.Methods The Sprague-Dawley rats underwent 30min of left anterior descending(LAD)coronary occlusion and 6 hours reperfusion to make ischemia/repefusion(I/R)injury model in vivo.Sixty male rats were randomly divided into 3 groups:sham group,I/R group,epigallocatechin-3-gallate group.Creatine kianse isoenzyme-MB(CK-MB)and the activity of Caspase-3 and the apoptotie index(AI)by TUNEL staining were measured in each group,I/R and EGCG group were measured the infarcted size(IS/AAR%).In addition,pathologic changes of myoeardial tissue were observed under electron microscopy.Results Compared with I/Rgroup,EGCG group markedly decreasedthe activity of CK-MB in serum[(951.57±123.71)vs(1826.38±205.32),P＜0.01]and the activity of Caspase-3 in myocardiaI tissue[(0.56±0.17)vs(0.81±0.20),P＜0.01],the value of IS/AAR% in EGCG group was lower than that in I/R group[(26.73±5.22)vs(41.56±6.81),P＜0.01].AI were significantly decreased in EGCG group compared with I/R group[(7.39±2.43)vs(15.62±4.28),P＜0.01].The electron microscopic examination showed that pathologic changes of myocardiocytes in the EGCG group were significantly milder than that of the I/R group.Conclusion Epigallocatechin-3-gallate has protective effect against myocardial ischemia and reperfusion injury in rats,and the protective mechanism may be related to decreasing the cardiomyocytes apoptosis by inhibition the activity of Caspase-3.

Objective To investigate the change of transmembrane potential during Noradrenaline's early preconditioning phase and probe the mechanism of action.Methods Primary culture myocardial cell of neonate rat were divided into three groups at random.GroupⅠ is control group,Group Ⅱis treatment group by hypoxia,Group Ⅲ is treatment group by NA.Group Ⅰ didn't give any drug to culture for 40minutes;Group Ⅱ treated for 40minutes by hypoxia,Group Ⅲ exposed to Noradrenaline(0.2?g/ml)for 10 minutes then treated with hypoxia for 40 minutes.The fluorescence intensity of rhodamine-123 marked mitochondria were detected by flow cytometry.Results The fluorescence intensity of three Group were test by analysis of variance(F=461.3,P

This study examined the effect of artesunate (Art) on the proliferation, DNA replication, cell cycles and apoptosis of vascular smooth muscle cells (VSMCs). Primary cultures of VSMCs were established from aortas of mice and artesunate of different concentrations was added into the medium. The number of VSMCs was counted and the curve of cell growth was recorded. The activity of VSMCs was assessed by using MTT method and inhibitory rate was calculated. DNA replication was evaluated by [3H]-TdR method and apoptosis by DNA laddering and HE staining. Flowmetry was used for simultaneous analysis of cell apoptosis and cell cycles. Compared with the control group, VSMCs proliferation in Art interfering groups were inhibited and [3H]-TdR incorprating rate were decreased as well as cell apoptosis was induced. The progress of cell cycle was blocked in G0/G1 by Art in a dose-dependent manner. It is concluded that Art inhibits VSMCs proliferation by disturbing DNA replication, inducing cell apoptosis and blocking cell cycle in G0/G1 phase.
Aorta/cytology ; Apoptosis/*drug effects ; Artemisinins/*pharmacology ; Cell Cycle/drug effects ; Cells, Cultured ; DNA Replication/*drug effects ; Muscle, Smooth, Vascular/*cytology ; Sesquiterpenes/*pharmacology

This study examined the effect of artesunate (Art) on the proliferation, DNA replication, cell cycles and apoptosis of vascular smooth muscle cells (VSMCs). Primary cultures of VSMCs were established from aortas of mice and artesunate of different concentrations was added into the medium. The number of VSMCs was counted and the curve of cell growth was recorded.The activity of VSMCs was assessed by using MTT method and inhibitory rate was calculated.DNA replication was evaluated by [3 H]-TdR method and apoptosis by DNA laddering and HE staining. Flowmetry was used for simultaneous analysis of cell apoptosis and cell cycles. Compared with the control group, VSMCs proliferation in Art interfering groups were inhibited and [3H]-TdR incorprating rate were decreased as well as cell apoptosis was induced. The progress of cell cycle was blocked in G0/G1 by Art in a dose-dependent manner. It is concluded that Art inhibits VSMCs proliferation by disturbing DNA replication, inducing cell apoptosis and blocking cell cycle in G0/G1 phase.

To examine the protective effect of insulin on reoxygenation-induced injury and explore the underlying mechanisms, the model of anoxia/reoxygenation (A/R) injury was established by inducing anoxia for 2 h and reoxygenation for 4 h in cultured cardiomyocytes of neonatal rats. The rats were randomized to four groups receiving vehicle, insulin, LY294002, insulin plus LY294002at the onset of reoxygenation after 2 h of anoxia. At the end of reoxygenation of 4 h, activity of lactate dehydrogenase (LDH) and content of malondialdehyde (MDA) were spectrophotometrically determined, apoptosis of cardiomyocytes were detected by using TUNEL and DNA Ladder, and Western blotting was employed to examine the expression of phosphorylated Akt in all groups. Our results showed that compared with vehicle-treated group, activities of LDH, contents of MDA, apoptosis index (AI) were significantly decreased, and expression of phosphorylated Akt was in creased significantly in insulin-treated group. However, changes in LDH, MDA, AI and phospho rylated Akt resulting from insulin were attenuated or abolished by LY294002 (PI3K inhibitor).These data strongly suggest that early administration of insulin at reoxygenation protects cardiomyocytes from reoxygenation-induced apoptosis through PI3K/Akt signaling pathway.

Aim To determine the effects of sodium tashinoneⅡA sulfonate(TSN) on monophasic action potential(MAP) and tachycardia-induced electrical remodeling of rabbit atria in vivo.Methods Twenty-four rabbits were equally divided into two groups randomly: control group and TSN group.Electrical catheters were localized in the right atrium through right internal jugular vein.Right atrial MAP was recorded by multiple channel recording. ERP of right atrial(AERP) was assessed by programmed electrical stimulation before pacing and from 0～8 hours after the onset of the pacing.Results The AERP_(200 ms) of control group was shortened from (105.9?3.8) ms to(114.7?7.2) ms and the rate-dependent of control group's atrium was lost through the pacing process compared with TSN group before pacing(P

Objective To study the effect of homocysteine(Hcy) on the expression of c-fos and c-jun mRNA and neointimal hyperplasia in rat common carotid artery, and to explore the possible mechanism of homocysteine exacerbating neointima formation after balloon injury. Methods The mRNA expression of c-fos and c-jun were detected by semi-quantitative RT-PCR in the common carotid artery. The morphology of arterial intima and media was studied by optical microscopy and image analysing system. Results The mRNA expression of c-fos and c-jun was significantly elevated in the low methionine(LM)〔(1.40?0.21),(1.43?0.25)〕 and high methionine(HM) 〔(1.68?0.27),(1.71?0.30)〕 than that in the control group 〔(1.10?0.15),(1.00?0.13), P

AIM: To evaluate whether tolerogenic dendritic cells (DC) loaded with heat shock protein 60 (HSP60) inhibit the progression of aortic atherosclerotic plaque in hypercholesterolemic apolipoprotein E (Apo-E) -null mice. METHODS: Bone marrow derived DC of the mice were loaded with HSP60 and co-cultured with rapamycin to generate tolerogenic DC. The tolerogenic DC, DC loaded only HSP60 and PBS were injected into the ApoE-null mice at 8 weeks of age for three times at a one-week interval. 8 weeks after the last injection, aorta were harvested for HE staining and anti-CD4~+T cell immunostaining. Responses of pleenic cells to HSP60 were also evaluated. RESULTS: Compared with DC, DC_ HSP60 expressed higher levels of CD86, and stimulated T lymphocytes to proliferation significantly, while the tolerogenic DC expressed lower levels of CD86, and inhibited T lymphocytes to proliferation. After immunization with different injection, the numbers of CD4~+ T cells in plaque were increased significantly in DC_ HSP60 group vs in PBS group (P