Objective: To observe the effects of Chinese patent medicines with the function of replenishing qi to activate blood (RQAB) plus ischemic postconditioning (IPoC) in protecting myocardium of rats from ischemia-reperfusion (I/R) injury, and to explore the possible mechanisms. Methods: Seventy-five Sprague-Dawley rats were randomly divided into sham-operated group (the suture was penetrated around the left anterior descending coronary artery, but not tied, n=15), I/R group (30 minutes of in situ transient occlusion of the left anterior descending artery, followed by 1 hour of reperfusion, n=15), IPoC group (30 minutes occlusion of the left anterior descending artery, followed by 3 cycles of 10 s of reperfusion/10 s of ischemia before 1-hour reperfusion, n=15), RQAB plus IPoC group (pretreated with 0.162 g/kg Xinyue Capsule and 0.135 g/kg Xiongshao Capsule for 14 days, and treated with IPoC 2 h after the final gavage, n=15), fosinopril sodium plus IPoC group (pretreated with fosinopril sodium, 0.9 mg/kg, n=15). Serum creatine kinase-MB (CK-MB) activity and cardiac troponin T (cTnT) level were detected; myocardial infarction size was measured by nitrotetrazolium blue chloride staining; Toll-like receptor 2 (TLR2) and TLR4 in myocardial tissue were examined by immunohistochemical method; interleukin-1beta (IL-1beta) and interleukin-6 (IL-6) levels in myocardial tissue were examined by enzyme-linked immunosorbant assay. Results: Compared with the I/R group, myocardial enzymes and infarction size were decreased significantly in the IPoC group (P<0.01); expressions of TLR2 and TLR4 and levels of IL-1beta and IL-6 in myocardial tissues were also significantly lower than those in the I/R group (P<0.05). Compared with the fosinopril sodium plus IPoC group, expressions of TLR2 and TLR4 were decreased significantly in the RQAB plus IPoC group (P<0.05, P<0.01). Compared with IPoC, RQAB plus IPoC reduced the infarction size and the release of myocardial enzyme CK-MB (P<0.01), and decreased the expressions of TLR2 and TLR4 and the levels of IL-1beta and IL-6 (P<0.05, P<0.01) in myocardial tissues. Conclusion: Pretreatment with Chinese herbs for nourishing qi and activating blood circulation can enhance the protective effect of IPoC on rat myocardial I/R injury, and its mechanism may be related to inhibition of TLR expression and expressions of the downstream proinflammatory cytokines.

This experiment was designed to search and identify the active principle as well as the optimal proportion of water-soluble extractives of traditional Chinese medicine (water-soluble extractives) Liqi Huoxue medicinals com-patibility (Qixue Bingzhi Fang-CWQB) in the prevention and treatment of atherosclerosis (As) by optimal uniform design method. The water-soluble extractives of CWQB were divided into 6 sections (A, B, C, D, E, F) through macroporous resin. The effect intensity and step of every component were compared through its effect on blood fat level, platelet aggregation, inflammatory factors, vascular endothelial growth factor (VEGF) and so on among hyper-lipoidemia rat models. The pharmacological experimental results and statistical analysis showed that CWQB water-soluble extractives of component D (mainly is paeoniflorin, accounted for 49.12%), component E (mainly is total flavonoids, accounted for 30.0%) compatibility had better effects on decreasing blood fat and triglyceride (TG). Com-pared with the model group, there was significant difference (P < 0.01). It also had inhibiting effect on endothelin (ET) and maximum platelet aggregation rate (P < 0.01). The component F (mainly is total acids, accounted for 32.7%) had inhibiting effect on serum IL-6 and IL-8 (P< 0.01). It was concluded that different compatibility of wa-ter-soluble extractives of CWQB can be applied to different targets or steps of the body. The active principle extrac-tives include main component of paeoniflorin, flavonoids and total acids. The best proportion is about 1:1:1.

Objective: in order to provide rapid and reliable method. Methods: Encoded Annexin Ⅴ cDNA was amplifyed from U937 cDNA libary by PCR and then subcloned into E coli expression vector. MS2-Annexin Ⅴ fusion protein could be overexpressed in E　coli. The MS2 bacteria protein could be removed by thrombin digestion.The mature Annexin Ⅴ was obtained by ion exchange chromatography and the FITC labled Annexin Ⅴ could be used in the detection of apoptosis. Results:Up to 37% of the total bacterial proteins was rhAnnexin Ⅴ as showed by SDS-PAGE. The purification of Annexin Ⅴ is over 99%. The FITC labled Annexin Ⅴ could efficiently detect apoptosis. Conclusion: We successfully established the technique procedure of obtaining a large quantity of Annexin Ⅴ and provided the basic routine for popularizing the detection of apoptosis' with high effciency.

OBJECTIVETo separate and characterize aqueous extracts of Salvia miltiorrhiza and Carthamus tinctorius to efficient, high-throughput and strong polar components, to observe effects of their aqueous effective components compatibility on rat myocardial ischemic reperfusion injury.

METHODMyocardial ischemic reperfusion injury model were established on SD rats by 40 min ligation of the left anterior descending artery and 120 min reperfusion. The rats were injected experimental drugs intravenously from femoral vein after 10 min ischemia. Rats were randomly divided into sham group (the suture around the left anterior descending coronary artery was not tied), model group, Danhong injection group (content of protocatechualdehyde is 0.05 g x L(-1), injection dosage equivalent to 1.80 g x kg(-1)), aqueous effective component of S. miltiorrhiza group (content of salvianolic acid B is 49 g x L(-1), injection dosage equivalent to 30.68 g x kg(-1)), aqueous effective component of S. miltiorrhiza group (content of hydroxysafflor yellow A is 31.76 g x L(-1), injection dosage equivalent to 17.87 g x kg(-1)), aqueous effective components compatibility of S. miltiorrhizae and C. tinctorius group (injection dosage is respectively 24.28 g x kg(-1) and 48.55 g x kg(-1)), each group have ten rats. Drugs were diluted with an equal dose of normal saline. The rats of sham group and model group were injected equivalent dosage of saline. The myocardial infarction size and the contents of serum cTnT and CK-MB were detected. The level of TXB2, 6-keto-PGF(1alpha) and platelet aggregation in blood plasma were investigated.

RESULTCompared with sham group, serum cTnT and CK-MB contents in model group increased significantly (P < 0.01). Compared with model group, myocardial infarction size and serum cTnT and CK-MB contents in aqueous effective component of S. miltiorrhiza group, aqueous effective component of C. tinctorius group and aqueous effective components compatibility of S. miltiorrhiza and C. tinctorius groups decreased significantly. Aqueous effective component of S. miltiorrhiza increased the level of 6-ke-to-PGF(1alpha), as well as decreased content of TXB2 and inhibited platelet aggregation (P < 0.01). Aqueous effective component of C. tinctorius also decreased the content TXB2 (P < 0.01). Improved extent of some detected markers in aqueous effective components compatibility of S. miltiorrhiza and C. tinctorius groups were better than that of Danhong injection group.

CONCLUSIONEffective components compatibility of aqueous extracts from S. miltiorrhiza and C. tinctorius may reduce myocardial infarct size and leakage of myocardial enzyme, and increase the level of 6-keto-PGF1alpha, so as to inhibit platelet aggregation and prevent thrombosis, the result of which is to reduce myocardial ischemic reperfusion injury.

Animals ; Carthamus tinctorius ; chemistry ; Disease Models, Animal ; Drug Interactions ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Myocardial Reperfusion Injury ; drug therapy ; Rats ; Rats, Sprague-Dawley ; Salvia miltiorrhiza ; chemistry

This study was aimed to observe the protective effects of components of water extract from Qi-Xue Bing-Zhi recipe (CWQB) on myocardial cells from hypoxia/reoxygenation (H/R) injury,by optimizing the metabolism of highenergy phosphates and then preventing cell damage and early apoptosis correlatively.Myocardial cells were separated and extracted from newborn SD rats.And then,H/R models were made by depriving oxygen for 3 hours and then regaining for 2 hours.Cardiomyocytes were divided into four groups,which were the control (normal oxygen) group,H/R (H/R model) group,TMZ (H/R model + 100 μmol/L Trimetazidine,TMZ) group,and CWQB (H/R model + 1 mmol/L CWQB) group.High-performance liquid chromatography (HPLC) was used in the content determination of adenosine monophosphate (AMP),adenosine diphosphate (ADP),adenosine triphosphate (ATP) in each group for the calculation of total adenylic acid (TAN) and energy charge (EC).Enzyme linked immunosorbent assay (ELISA) was used in the detection of myocardial damage markers,such as creatine kinase-MB (CK-MB) and cardiac troponin T (cTnT).The early apoptosis of myocardial cells were assessed by flow cytometry (FCM).The results showed that compared with H/R group,contents of AMP in the TMZ group and CWQB group were decreased,while contents of ADP,ATP,TAN and EC were all increased in both groups.The increasing degrees of ADP,ATP and TAN in TMZ group were higher than those of the CWQB group (P ＜ 0.05).Contents of CK-MB and cTnT were significantly decreased in the TMZ group and the CWQB group.Content of cTnT decreased more significantly in the CWQB group (P ＜ 0.05).In the TMZ group and the CWQB group,early apoptosis rate was decreased.The decreasing in the TH group was more significant (P ＜ 0.05).The correlation analysis showed that the level of CK-MB and concentration of ADP had significant negative correlation.The early apoptosis rate and AMP had significant positive correlation.The early apoptosis rate had negative correlation with concentrations of ADP and ATP (P ＜ 0.01).It was concluded that CWQB recipe can decrease the concentration of AMP in H/R cardiomyocytes,increase the concentrations of ADP,ATP,TAN and EC,and decrease myocardial damage makers such as CK-MB and cTnT,depress early apoptosis rate in H/R cardiomyocytes,in order to display its cardioprotective effects in H/R injury.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection can cause acute respiratory distress syndrome, hypercoagulability, hypertension, and multiorgan dysfunction. Effective antivirals with safe clinical profile are urgently needed to improve the overall prognosis. In an analysis of a randomly collected cohort of 124 patients with COVID-19, we found that hypercoagulability as indicated by elevated concentrations of D-dimers was associated with disease severity. By virtual screening of a U.S. FDA approved drug library, we identified an anticoagulation agent dipyridamole (DIP) , which suppressed SARS-CoV-2 replication . In a proof-of-concept trial involving 31 patients with COVID-19, DIP supplementation was associated with significantly decreased concentrations of D-dimers ( < 0.05), increased lymphocyte and platelet recovery in the circulation, and markedly improved clinical outcomes in comparison to the control patients. In particular, all 8 of the DIP-treated severely ill patients showed remarkable improvement: 7 patients (87.5%) achieved clinical cure and were discharged from the hospitals while the remaining 1 patient (12.5%) was in clinical remission.