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 The aim of this prospective observational study was to analyze the prevalence and the predictive factors of hemorrhagic events after abdominal paracentesis in patients with acute-on-chronic liver failure (ACLF).Methods ACLF patients who received at least one episode of abdominal paracentesis were prospectively enrolled between January 2010 to December 2013. Prevalences of intraperitoneal and abdomen hemorrhage complications were examined. t test was performed for continuous variables and chi-square test was performed for categorical variables.Binary Logistic regression was used to analyze the risk factors of hemorrhage.Results A total of 525 abdominal paracenteses were carried out on 185 ACLF patients within a 4-year period,with 289 (55 .0%)for diagnostic purpose and 236 (45 .0%)for therapeutic purpose.A total of 16 (3.0%)hemorrhagic complications were identified, with 4 cases of abdominal wall hematomas and 12 cases of intraperitoneal hemorrhage.Patients were divided into hemorrhage group and non-hemorrhage group according to this complication.Age,gender, Child-Pugh score,volume of ascitic fluid removed,underlying cirrhosis,platelet count and thrombin time were not significantly different between two groups (all P > 0.05 ).Patients with bleeding events had lower fibrinogen levels and higher prothrombin time,international normalized ratio,activated partial thromboplastin time and model for end-stage liver disease score (all P <0.05).After adjustment of other factors,multivariate regression analysis indicated that low fibrinogen level was the only independent predictor of hemorrhagic complication (OR=0.105,95%CI :0.018-0.608,P =0.012).Conclusion Low fibrinogen level is the independent predictor of severe hemorrhagic complications following paracenteses in patients with ACLF.

The practice of general practice emphasizes that the general practitioner is the backbone, and the primary health institution is the main body, which solves the main health problems of the commu-nity and meets the basic health service needs of the residents. Rehabilitation medicine occupies a prominent position in the community health service, and its related knowledge and skills are the important part of the whole medical education. However, rehabilitation medicine has more teaching contents, less training time and out-of-date teaching model, which have made the effect of rehabilitation medicine teaching not good. In the rehabilitation medicine teaching for training general practitioners, by strengthening the concept of reha-bilitation, elaborating teaching content, optimizing teaching ward-round, and outstanding ability as well as reforming evaluation way, we try to exercise the students' clinical thinking and improve clinical skills, to cultivate outstanding qualified general medical practitioners for community rehabilitation ultimately.

The present paper is aimed to explore the biological osteoinductive activity of recombinant human bone morphogenetic protein 9 (rhBMP-9) by various biological technologies. In this study, we firstly obtained hBMP-9 cDNA by PCR and inserted it into vector pcDNA4/His Max to reconstruct hBMP-9 eukaryotic expression vector pcDNA4/His Max-BMP-9. Recombinant Chinese hamster ovary (rCHO) cell line expressing high-level rhBMP-9 was reconstructed by co-transfecting the expression vectors pcDNA4/His* Max-hBMP-9 and plasmid pSV2-dhfr into dihydrofolate reductase (dhfr)-deficient CHO cells and the subsequent gene amplification by the methotrexate. We finally obtained a monoclonal cell line expressing the highest level protein. We purified the medium after culturing the highest-producing monoclonal by Ni-NTA His-Bind Resin columns and concentrated to by a Centricon 50 at 4 degrees C and stored at 70 degrees C until it was used. Western blot and SDS-PAGE analyses showed a specific band of about 32kD in pro-region lane and a specific band of about 50kD in pro-region complex lane. Biological activities of rhBMP-9 were tested by colorimetric determination and histochemical staining of Alkaline Phosphatase (ALP) Activity, osteocalcin and oesteopontin for C3H10 T1/2 cells, which were stimulated culture by different concentration (20, 50, 100 microg/mL) of rhBMP-9. The results showed that the rhBMP-9 could induce osteogenic differentiation of mesenchymal stem cells (MSCs) in vitro, and were proportional to the amount. This study can provide experimental data for further tests in vivo and clinical applications.
Animals ; CHO Cells ; Cricetinae ; Cricetulus ; Growth Differentiation Factor 2 ; biosynthesis ; isolation & purification ; pharmacology ; Humans ; Osteogenesis ; drug effects ; Recombinant Proteins ; biosynthesis ; isolation & purification ; pharmacology ; Transfection

OBJECTIVETo establish quality control criteria for medicinal herb Cajanus cajan based on the determination of longistylin A and longistylin C, two bioactive and specific stilbenes of the plant.

METHODLongistylin A and longistylin C were obtained from the leaves of C. cajan by silica gel column chromatography and identified as marker compounds of this plant by spectroscopic analysis. A RP-HPLC method was established to determine the two compounds.

RESULTLongistylin A and longistylin C were well separated on a Thermo BDS Hypersil C18 column (4.6 mm x 250 mm, 5 microm) with a mobile phase methanol-water (8:2), and showed good linearity in the range of 0.00288 - 0.0576 microg and 0.0112 - 0.224 microg, respectively. The average recoveries were 98.9% and 97.2% with RSD of 2.4% and 2.2% for these two compounds, respectively.

CONCLUSIONThe established analysis method is simple and accurate, whicn can be used for quality control of C. cajan.

Cajanus ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Diethylstilbestrol ; analysis ; isolation & purification ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Plant Leaves ; chemistry ; Plants, Medicinal ; 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.