This study was to investigate the protective effects of puerarin on myocardial ischemia/reperfusion (MI/R) injury and the underlying mechanism. The MI/R-model was established by ligating the left anterior descending artery (LAD) for 60 min followed by 24 h reperfusion, puerarin (10, 30, and 100 mg·kg^-1) was orally administered 20 min before reperfusion. Cardiac function, myocardial infarct index, cardiac damage markers, inflammatory cytokines, and apoptosis index were measured to evaluate the protective effects of puerarin on MI/R injury. The activation of Nod-like receptor protein 3 (NLRP3) inflammasome and Toll like receptor 4 (TLR4)/myeloid differentiation factor 88 (Myd88)/nuclear factor kappa B (NF-κB) pathway were determined by Western blot. All animal experimental procedures were approved by the ethics committee of the Institute of Materia Medica, Peking Union Medical College, Chinese Academy of Medical Sciences. The results showed that puerarin could significantly improve cardiac function, reduce myocardial infarct size, decease the levels of lactic dehydrogenase (LDH), aspartate transaminase (AST), creatine kinase-MB (CK-MB), and cardiac troponin T (cTnT) and suppress cardiomyocyte apoptosis. Meanwhile, puerarin could notably decrease the levels of inflammatory cytokines such as interleukin-1β (IL-1β), IL-6, and tumor necrosis factor-α (TNF-α). Western blot analysis revealed that puerarin could downregulate the expression of TLR4, Myd88, NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), cleaved-caspase 1, cleaved-gasdermin-D (GSDMD), IL-1β, and IL-18, as well as the phosphorylation levels of inhibitor of NF-κB α (IκBα), IκB kinase β (IKKβ), and NF-κB. These findings demonstrated that puerarin could alleviate MI/R injury by suppressing NLRP3 inflammasome activation, possibly via TLR4/Myd88/NF-κB pathway.

OBJECTIVETo study the changes of inducible nitric oxide synthase (iNOS) activity and apoptosis-related genes Bcl-2, Bax and caspase-3 mRNA expressions in endotoxemia-induced rat diaphragm injury and analyze the related apoptosis mechanism.

METHODSThirty-two male SD rats were randomly divided into 4 groups (n = 8): control group (saline 0.5 ml ip), endotoxin 24 h, 48 h and 96 h group (endotoxin 12 mg/kg ip, animals were killed either 24, 48 or 96 h after injections). Body weight were measured, the ratio between diaphragm weight and body weight, activities of constitutive nitric oxide syntheses (cNOS), iNOS and succinate dehydrogenase (SDH) were also measured. The expressions of Bcl-2, Bax and caspase-3 mRNA were detected by RT-PCR analysis.

RESULTSEndotoxin induced significant reductions in diaphragm mass in endotoxin 96 h group (P < 0.05). Endotoxin increased diaphragm cNOS or iNOS activities, and they were significantly higher in endotoxin 96 h group than those in endotoxin 24 h and 48 h groups, diaphragm SDH activity was reduced, and it was lower in endotoxin 96 h group than that in endotoxin 24 h and 48 h groups (P < 0.01). Endotoxin significantly increased Bax and caspase-3 mRNA expressions, and they were higher in endotoxin 48 h and 96 h groups than those in endotoxin 24 h group (P < 0.01). Endotoxin significantly reduced Bcl-2 mRNA expression and the ratio of Bcl-2/Bax, and they were lower in endotoxin 48 h and 96 h groups than those in endotoxin 24 h group (P < 0.01).

CONCLUSIONiNOS is activated in endotoxemia-induced rat diaphragm injury. It damages mitochondria, upregulates Bax expression and downregulates Bcl-2 expression, then induces caspase-3 related apoptotic pathway. These changes may cause diaphragm injury and atrophy.

Animals ; Apoptosis ; Caspase 3 ; metabolism ; Diaphragm ; metabolism ; physiopathology ; Endotoxemia ; metabolism ; Gene Expression ; Male ; Nitric Oxide Synthase Type II ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley ; bcl-2-Associated X Protein ; metabolism

Baculoviridae ; genetics ; metabolism ; Phylogeny ; Viral Proteins ; chemistry ; classification ; genetics ; metabolism

HPLC-ELSD was applied to explore the absorption mechanism of pulchinenosides (B3, BD, B7, B10, B11) in rats. The experimental results showed that the absorption rate constant, Ka value (B3, BD) and Permeability coefficient, Peff value (B3, B7) displayed significant difference (P <0.05) in various intestinal segments, The Ka value and Peff value of PRS was different from each other with the highest absorption in duodenum (duodenum > jejunum > colon > ileum); The PRS displayed excessive satuation as the concentration increased over 0.05-2.5 g · L(-1). There were no obvious linear correlations between Peff values and concentrations in duodenum (0.6007 ≤ R2 ≤ 0.7727); Ka and Peff value declined when the PRS was perfused with P-glycoprotein promoter digoxin, on the other hand, inclined when perfused with P-glycoprotein inhibitor verapamil with significant difference among PRS B3, BD, B7, B11 (P <0.05). All the above results demonstrated that B3, BD, B7 were greatly influenced by absorption sites, duodenum was the main absorption site; PRS didn't entirely transported in a concentration dependent manner, and the transporter-protein involved the transportation, so the intestinal absorption of the five pulchinenosides was not entirely passive diffusion; and PRS might be the substrates of P-glycoprotein.
ATP-Binding Cassette, Sub-Family B, Member 1 ; physiology ; Animals ; Intestinal Absorption ; Male ; Oleanolic Acid ; pharmacokinetics ; Pulsatilla ; chemistry ; Rats ; Rats, Wistar ; Saponins ; pharmacokinetics

Objective To study on the effects of Cinnamic aldehyde on leukemia cell line K562 by Caco -2 cells in vitro absorption model.Methods The effective components of cinnamon(0,50,100,200,400, 600,800,1000 μg· mL-1 ) were determined by Caco-2 cell model of Transwell, and the concentration was determined by HPLC.No cytotoxic concentration range of Cinnamic aldehyde acting on K562 cells for 72 h is detected by MTT assay.After 72 h incubation of Cinnamic aldehyde standard(50,75 μg· mL-1 ) and leukemia K562 cells, the cells surface antigens including CD235a, CD36, CD41, CD61, CD13, CD33 and CD14 were determined by Flow cytometry.Results The active ingredi-ent of cinnamon is extracted by transwell transport pool of Caco-2 cell model and no cytotoxic concentration is 200 μg · mL-1.The cinnamicaldehyde is the component which goes through the model by HPLC.The 24 h inhibition rates ( IRs ) of Cinnamic aldehyde on K562 cells are (25.29 ±0.97)%and (36.60 ±0.18)%at the concentrations of 50 and 75 μg· mL-1 , respectively;IRs for 48 h are ( 48.23 ±0.63 )% and ( 57.15 ±0.58 )%; IRs for 72 h are ( 58.23 ±0.63 )% and (57.15 ±0.58)%.Compared with the control group, the inhibitory activity is obvious(P<0.05).After incubation 72 h, the expressions of myeloid differentiation phenotypes including CD13, CD33, CD36 on K562 cells are (0.33 ±0.21)%, ( 32.89 ±0.19 )%, ( 7.73 ±0.57 )% and ( 0.72 ±0.43 )%, ( 38.80 ±0.03 )%, (10.90 ±0.82)%at the concentrations of 50 and 75 μg· mL-1 , respectively.Compared with the control group, the inhibition increased ( P <0.05 ).The phenotypic expressions of erythroid differentiation are ( 52.38 ±0.65 )%, (57.48 ±0.70)%.Compared with the control group, the inhibition increased( P<0.05).Megakaryocyte differentia-ted phenotype CD41, CD61 expression has no significant change ( P >0.05 ).Conclusion The Cinnamic aldehyde can go through the Caco-2 in vitro absorption model and enables the K562 cells to differentiate into myeloid and erythroid.

Mammalian oocyte maturation and early embryo development processes are Ca(2+)-dependent. In this study, we used confocal microscopy to investigate the distribution pattern of Ca2+ and its dynamic changes in the processes of bovine oocytes maturation, in vitro fertilization (IVF), parthenogenetic activation (PA) and somatic cell nuclear transfer (SCNT) embryo development. During the germinal vesicle (GV) and GV breakdown stage, Ca2+ was distributed in the cortical ooplasm and throughout the oocytes from the MI to MII stage. In IVF embryos, Ca2+ was distributed in the cortical ooplasm before the formation of the pronucleus. In 4-8 cell embryos and morulas, Ca2+ was present throughout the blastomere. In PA embryos, Ca2+ was distributed throughout the blastomere at 48 h, similar to in the 4-cell and 8-cell phase and the morula. At 6 h after activation, there was almost no distribution of Ca2+ in the SCNT embryos. However, Ca2+ was distributed in the donor nucleus at 10 h and it was distributed throughout the blastomere in the 2-8 cell embryos. In this study, Ca2+ showed significant fluctuations with regularity of IVF and SCNT groups, but PA did not. Systematic investigation of the Ca2+ location and distribution changes during oocyte maturation and early embryo development processes should facilitate a better understanding of the mechanisms involved in oocyte maturation, reconstructed embryo activation and development, ultimately improving the reconstructed embryo development rate.
Aniline Compounds/chemistry ; Animals ; Calcium/*physiology ; Cattle/*physiology ; Embryonic Development/*physiology ; Female ; Fertilization in Vitro/*veterinary ; Microscopy, Confocal/veterinary ; Oocytes/*physiology ; Parthenogenesis/*physiology ; Xanthenes/chemistry

Angiogenic gene therapy and cell-based therapy for peripheral arterial disease(PAD) have been studied intensively currently. This study aimed to investigate whether combining mesenchymal stem cells(MSCs) transplantation with ex vivo human hepatocyte growth factor(HGF) gene transfer was more therapeutically efficient than the MSCs therapy alone in a rat model of hindlimb ischemia. One week after establishing hindlimb ischemia models, Sprague-Dawley(SD) rats were randomized to receive HGF gene-modified MSCs transplantation(HGF-MSC group), untreated MSCs transplantation (MSC group), or PBS injection(PBS group), respectively. Three weeks after injection, angiogenesis was significantly induced by both MSCs and HGF-MSCs transplantation, and capillary density was the highest in the HGF-MSC group. The number of transplanted cell-derived endothelial cells was greater in HGF-MSC group than in MSC group after one week treatment. The expression of angiogenic cytokines such as HGF and VEGF in local ischemic muscles was more abundant in HGF-MSC group than in the other two groups. In vitro, the conditioned media obtained from HGF-MSCs cultures exerted proproliferative and promigratory effects on endothelial cells. It is concluded that HGF gene-modified MSCs transplantation therapy may induce more potent angiogenesis than the MSCs therapy alone. Engraftment of MSCs combined with angiogenic gene delivery may be a promising therapeutic strategy for the treatment of severe PAD.
Animals ; Bone Marrow ; metabolism ; pathology ; Bone Marrow Transplantation ; Cells, Cultured ; Hepatocyte Growth Factor ; genetics ; Hindlimb ; pathology ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; metabolism ; pathology ; Neovascularization, Physiologic ; genetics ; Rats

OBJECTIVETo summarize the clinical characteristics of secondary coagulation disorders caused by exposure to poison (raticide) in children and to investigate the diagnosis and corresponding treatment.

METHODThe process of diagnosis, clinical characteristics, response to treatment and the prognosis were analyzed.

RESULTSThe main clinical manifestation was mucosal bleeding (66.6%), including epistaxis, gingival bleeding, hematomas and so on. All these children were previously well and had no history of bleeding. Activated partial thromboplastin time (APTT) and prothrombin time (PT) were prolonged, factor II was undetectable and the levels of factors VII, IX, and X were lower. The fibrinogen was normal. A raticide was detected in blood and urine of 13 children although 12 of the patients had no definite history of raticide ingestion. Prothrombin complex, fresh frozen plasma and vitamin K(1) were effective in these cases. However, 2 - 3 weeks later, 6 patients presented with recurrent bleeding.

CONCLUSIONFor children with secondary coagulation disorders of unknown cause, intoxication of raticide should be considered. The administration of blood coagulation factors and vitamin K(1) are effective in early treatment, and the treatment period should be more than 2 months. The PT and APTT should be followed up. Vitamin K(1) should be stopped when PT and APTT are normal.

Blood Coagulation Disorders ; chemically induced ; diagnosis ; therapy ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Rodenticides ; poisoning ; Vitamin K 1 ; administration & dosage ; therapeutic use

OBJECTIVESTo evaluate the effect of Happy 10 program on the promotion of physical activity, physical growth and development of primary-school students, and on obesity control and prevention.

METHODSTwo similar primary schools from one district of Beijing, China were selected, one as an intervention school and the other as a control school. Happy 10 program was implemented at least once every school day in the intervention school for two semesters, whereas no intervention was adopted in the control school. The information on energy expenditure and duration of physical activity was collected by a validated 7-day physical activity questionnaire. Height and weight were measured by trained investigators following the standardized procedure. Energy expenditure and intensity of each Happy 10 session was measured by a physical activity monitor.

RESULTSThe average energy expenditure and duration of total physical activity per day among students in the intervention school increased significantly from 15.0 to 18.2 kcal/kg, and 2.8 to 3.3 h respectively, whereas the figures significantly decreased in the control school. There was a significant difference in change of weight and BMI between girls in the intervention and control school (2.4 kg vs 4.6 kg, -0.47 kg/m2 vs 0.66 kg/m2). The prevalence of overweight and obesity in the intervention school decreased by 0.4%-5.6%, as compared to the increase by 0.6%-4.5% in the control school. The average energy expenditure and intensity per 10-minute session ranged from 25.0-35.1 kcal, 4.8-6.2 kcal/kg/h respectively in grades 1-5.

CONCLUSIONHappy 10 program provides a useful strategy to promote physical activity among school children and also plays a positive role in building up physical growth and development of girls.

Body Height ; Body Weight ; Child ; China ; Energy Metabolism ; Exercise ; physiology ; Female ; Government Programs ; Health Promotion ; methods ; Humans ; Male ; Obesity ; prevention & control ; Physical Fitness ; physiology ; Schools

Adult ; Aged ; Female ; Femoral Fractures ; surgery ; Femur ; injuries ; surgery ; Fracture Fixation, Internal ; methods ; Fractures, Ununited ; surgery ; Humans ; Male ; Middle Aged ; Young Adult