Purpose To study on the antithrombotic effect of polysaccharide of Auricularia auriculajudae.Methods The method of hot water extraction for extracting polysaccharide from Auricularia auriculajudae was used. The antithrombotic effect was studied by animal experiments.The length and the weight of arterial thrombosis were determined. Characteristic thrombus formation time (CTFT), and thrombus formation time (TFT),the rate of platelet adhesion and the blood viscosity were determined also.Results As compared with the normal saline group,polysaccharide of Auricularia auriculajudae(ig) remarkedly prolonged CTFT and TFT.The length of thrombus was shortened. Thrombus wet weight, thrombus dry weight,and blood viscosity were decreased, yet had no significant influence on the rate of platelet adhesion.Conclusion Polysaccharides of Auricularia auriculajudae can significantly inhitit the formation of thrombus.

Purpose To test the virus inactivation effect of water bath method at 60 ℃ for 10 hours and alcohol treatment for 3 hours which was used in the production of urinary trypsin inhibitor(UTI).Methods Sindbis virus,Pseudorabies virus(PRV) and poliovirus1(PV1) were used as indicated viruses in this test.After being added separately into the UTI raw material in 10% proportion,the viruses were treated with water bath at 60 ℃ for 10 hours and alcohol for 3 hours and then the samples of UTI were taken to inoculate the cell line for assay of cytopathic effect.Results The water bath at 60 ℃ for 10 hours could inactive Sindbis,PRV and PV1 in more than(6.503±0.102)LgTCID_(50),(6.42±0.158) LgTCID_(50) and(6.587±0.061)LgTCID_(50) respectively,and alcohol treatment for 3 hours could inactive Sindbis,PRV and PV1 in more than(5.88±0.204)LgTCID_(50),(6.378±0.268)LgTCID_(50) and(5.963±0.118) LgTCID_(50) respectively.No cytopathic effect was found in the cell line which was inoculated with treated samples after blind passage for three generations.Conclusion The water bath method at 60 ℃ for 10 hours and alcohol treatment for 3 hours which were used in the production of UTI had good effects on virus inactivation and the inactivation efficiency on Sindbis,PRV and PV1 was more than 6 LgTCID_(50)/mL.

Purpose To observe the effects of umbilical cataplasm on rat gastrointestinal motility and gastrointestinal hormones using rat model of gastric operation,and to explore its possible mechanism.Methods ①The peristalsis length of carbon ink in small intestine of 20 rats was investigated by the use of umbilical cataplasm,compared with the matrix cataplasm group and the untreated control group. ②By means of radioimmunoassay,the levels of motilin(MOT) and somatostatin(SS) in small intestine tissue of 20 rats were investigated after a ten-day course of application of umbilical cataplasm.Results ①The group of umbilical cataplasm could significantly increase the peristalsis length of carbon ink in the rat small intestine and the distance percentage to the whole intestine,compared with the matrix cataplasm group and the untreated control group has significant difference(P＜0.01),and has no significant differernce between the other two groups(P＞0.05). ②Umbilical cataplasm can increase the levels of MOT and decrease the levels of SS in small intestine tissue of rat,compared with the untreated group P＜0.05 or P<0.01.Conclusion Umbilical cataplasm can promote gastrointestinal peristalsis of the rat model of gastric operation, and can regulate the levels of gastrointestinal hormones.

Purpose To investigate the effect of small interfering RNA on the increase of myocyte enhancer factor 2A(MEF2A) expression in PC12 cells exposed to hypoxia.Methods PC12 cells were cultured under normal conditions or were exposed to hypoxic conditions.Small interfering RNA targeted MEF2A gene (MEF2A-siRNA) was chemically synthesized. Eukaryocytic expression vector was built and transfected into PC12 cells with liposome. The expression of MEF2A mRNA was detected by real-time PCR. Western blot was used to detect the MEF2A protein.Results Compared with normal control(2~(-△△CT)=1.01±0.02), the mRNA level of MEF2A gene in PC12 cells with the treatment of MEF2A-siRNA was down-regulated significantly by 88%(2~(-△△CT)=0.12±0.03, P＜0.01).The expression of MEF2A protein in hypoxia-treated PC12 cells was markedly higher than that of normal control(98.4±11.7 and 47.5±7.6,P＜0.01).However, MEF2A-siRNA could significantly suppress the increase of MEF2A protein (P＜0.01).Conclusion MEF2A gene silence induced by siRNA might inhibit the increase of MEF2A protein by hypoxia in PC12 cells.

Purpose To explore the relationship between connective growth tissue factor(CTGF) in serum and the severity of liver fibrosis,and to determine the clinical value of CTGF in the assessment of liver fibrosis.Methods Serums CTGF were tested utilizing enzyme linked immunosorbent assay(ELISA).The correlation between serum CTGF concentration and fibrosis stage was assessed.Results The diagnostic performance of CTGF was assessed by comparing the area under receiver characteristic curves(AUC) with a panel of fibrosis markers.Correlation coefficient was 0.689(P＜0.001) between the levels of serum CTGF and fibrosis stages and AUC of CTGF was 0.841(95% confidence interval,0.762-0.920) in distinguishing mild fibrosis from significant fibrosis.Conclusion The present data revealed that serum CTGF was significantly correlated with the stage of liver fibrosis,suggesting that serum CTGF was an indicator for the stage of liver fibrosis,and serum CTGF could be used as a valuable marker assessing liver fibrosis.

Purpose To investigate the effects and the mechanism of oleum curcumae wenchowensis (OCW) with different concentrations (0, 2.5, 5, 10 and 20 mg/mL) on chronic myeloid leukemia cell line K-562 cells in vitro.Methods The apoptosis of K-562 cells was dyed by Hoechest 33258 and detected by flow cytometry marked with Annexin V/PI. The Expression of Fas/FasL, bcr/abl, bcl-2 and p53 was detected by semi-quantity reverse transcript-polymerase chain reaction (RT-PCR) and Western blot.Results The results showed that the apoptosis rates were gradually elevated. The expression of Fas and FasL protein was increased in a concentration dependent manner, while bcr/abl, bcl-2 and p53 had no significant changes.Conclusion OCW could induce the apoptosis of K-562 cells by up-regulating the expression of Fas/FasL protein.

Purpose To examine the regulatory effect of recombinant human fibroblast growth factor-21 on the expression of liver X receptor α and glucose transporter protein 1 in the type 2 diabetes mellitus rats.Methods The rat models of type 2 diabetic mellitus were divided into four groups at random, ic. rhFGF-21 every day, after eight weeks of these treatment, Inspect the fasting blood glucose (FBG), fructosamine(FA), triglyceride(TG), T-cholesterol(TC), high density lipoprotein cholesterol(HDL-C) and low density lipoprotein cholesterol(LDL-C) of these rats, then detecting the mRNA expression of LXRα and GLUT1 by RT-PCR.Results (1) rhFGF-21 can reduce blood glucose steadily to near normal levels in diabetic rats. (2) The expression of LXRα and GLUT1 level was significantly higher in the rhFGF-21 treatment group than that in the model group. (3) rhFGF-21 megadoses and middle doses decreased FA, TG, TC,and LDL-C and elevated HDL-C.Conclusion rhFGF-21 could regulate the mRNA expression of LXRα and GLUT1 in diabetes rats, increase basal level glucose transport, then reduce blood glucose, improve lipid metabolize dysfunction.

Purpose To compare pharmacokineics of puerarin and crude extract in rats.Methods Rats received 500 mg/kg puerarin and puerarin crude extract by oral administration respectively.Hydroxybenzoic acid was selected as internal standard and the plasma concentration of the puerarin and crude extract was analyzed by HPLC.The pharmacokinetics parameters were calculated with DAS2.0.Results The pharmacokinetics of puerarin and puerarin crude extract was both best fitted with two-compartment models in rats after oral administration,and the pharmacokinetics main parameters of the two formulations were different:the AUC_(0-t) and C_(max) of puerarin were much greater than those of puerarin crude extract,but T_(max),t_(1/(2z)),CL/F and V_z/F were much lesser than those of puerarin crude extract.Conclusion The complex components in pueraria crude extract can affect the pharmacokinetics of puerarin in rat in vivo.

Purpose To investigate the effects of oxytocin(OT) and prostaglandins intervention on oxytocin receptor(OTR) expression of primary culture of human myometrial smooth muscle cell.Methods Using respectively oxytocin, prostaglandin E2(PGE2),and prostaglandin F2alpha(PGF2alpha) intervened on human myometrial smooth muscle cell.The expression of OTR mRNA and protein of cell hemogenate was examined.Results The expression of OTR between oxytocin intervention group and untreated group was similar.The expression of OTR in cell was significantly higher in the PGE2 or PGF2alpha intervention group than that in the untreated group.The expression of OTR in cell was significantly higher in the PGE2 and PGF2alpha joint intervention group than that in the untreated group and than that in the PGE2 or PGF2alpha individual intervention group.Conclusion Oxytocin didn′t increse the expression of OTR in human myometrial smooth muscle cell.PGE2 and PGF2alpha incresed the expression of oxytocin receptor in human myometrial smooth muscle cell.Furthermore PGE2 and PGF2alpha joint intervention more significantly increased the expression of oxytocin receptor than PGE2 or PGF2alpha individual intervention in human myometrial smooth muscle cell.

Purpose A RP-HPLC method was used to determine genioside and breviscapine in plasma and to study its pharmacokinetics in rat, respectively.Methods Rat plasma samples were collected after a single dose of Zhideng injection and pharmacokinetic parameters of genioside and breviscapine were estimated,respectively.Results A good linear relationship was obtained between 0.2-40.0 μg/mL for breviscapine, and 0.5-200.0 μg/mL for genioside.The recoveries from plasma were larger than 85%,and RSDs of inter-day asaay and intra-day assay were below 10%. The pharmacokinetic results showed that genioside and breviscapine were rapidly eliminated from plasma after iv administration of three doses of Zhideng injection.The mean half-life was 72.6 min and 21.6 min,respectively.Conclusion The established HPLC method was suitable for the pharmacokinetic study of genioside and breviscapine.