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OBJECTIVE:To investigate the e ffects of Zhuangtongyin water extract (ZTYWE)on hemorheology and blood lipid level of myocardial ischemia-reperfusion model rats ,and preliminarily explore its molecular biology foundation of reducing arrhythmia after myocardial ischemia-reperfusion. METHODS :Totally 75 SD rats were divided into control group ,model group and ZTYWE low-dose ,middle-dose and high-dose groups. Myocardial ischemia-reperfusion model was established by ligating the left ventricle cyclotron endings for 30 min then reperfusing for 60 min. Since the first day after surgery ,control group and model group were intraperitoneally administrated with same volume normal saline ;ZTYWE low-dose ,middle-dose and high-dose groups were intraperitoneally administrated with medicine solution (6.8,13.6,27.2 g/kg),once a day ,for continuous 28 d. At the 7th, 14th,21st,28th day after surgery ,the incidence rate of arrhythmia of rats were detected. The hemorheology indicators and serum contents of TC ,TG,LDL-C,HDL-C were determined after the last administration. The morphology of myocardial tissue was observed by HE staining. The protein expressions of apoptosis-related factor FAS and FAS-L were detected by Western blotting assay. RESULTS :Compared with control group ,the incidence rates of arrhythmia of rats in model group were significantly increased;the whole blood contrast viscosity ,hematocrit(HCT),in vitro forming length and quality of thrombosis ,platelet aggregation rate ,erythrocyte filtration index (IF),as well as the serum contents of TC ,TG and LDL-C were significantly increased;in vivo thrombus formation time as well as the serum content of HDL-C were significantly decreased ;the protein expression of FAS and FAS-L were significantly increased (P<0.05 or P<0.01);obvious pathological changes were observed in myocardial tissue. Compared with model group , the incidence rates of arrhythmia of rats in ZTYWE groups were in obvious reducing trend ; except for the HCT in 163.com low-dose ZTYWE group without significance , the blood contrast viscosity ,HCT,in vitro forming length and quality of thrombosis ,platelet aggregation rate ,IF as well as the serum contents of TC ,TG and LDL-C were significantly decreased ;in vivo thrombus formation time as well as the serum contents of HDL-C were significantly increased ;the protein expressions of FAS and FAS-L were significantly decreased (P<0.05 or P< 0.01);the pathological changes of myocardial tissue were improved. CONCLUSIONS :Through the promoting blood circulation and removing blood stasis effect ,Zhuangtongyin decoction can improve indexes of hemorheology index of ischemia-reperfusion model rats ,reduce the contents of TC ,TG and LDL-C ,enhace the contents of HDL-C ,down-regulate the protein expression of FAS and FAS-L ,so as to inhibit myocardial cells apoptosis and reduce arrhythmia.
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OBJECTIVE: To observe the effects of Zhuangtongyin (ZTY) water extract on blood vessels diastolic and systolic function and cardiac function in rats with coronary heart disease (CHD) and its potential mechanism. METHODS: SD rats were randomly divided into control group, sham operation group, model group, positive group (Compound danshen dripping pills, 0.08 g/kg) and ZTY water extract low-dose, medium-dose and high-dose groups (6.8, 13.6, 27.2 g/kg, by weight of water extract), with 10 rats in each group. Except for control group (no operation) and sham operation group (only open chest without ligation), CHD model were set up in other groups by ligating the left ventricle cyclotron endings. One week after modeling, control group, sham operation group and model group were given constant volume of water intragastrically, and administration groups were given relevant medicine intragastrically; once a day, for consecutive 4 weeks. Two hours after last administration, cardiac function indexes (LVEDV, LVESV, SV, LVEF) of rats were detected in each group. The levels of inflammatory factors (CRP, IL-1β, IFN-γ and ET-1) in myocardial tissue were determined by ELISA. Histomorphological characteristics of myocardial tissue were determined by HE staining. RESULTS: The structure of myocardial fibers in control group and sham operation group was clear and orderly; there was no statistical significance in difference of each index between 2 groups (P>0.05). Compared with control group, myocardial fibers were disorderly arranged in model group, and the phenomena of rupture, dissolution and necrosis were observed, accompanied by infiltration of inflammatory cells. The levels of LVEDV and LVESV as well as the levels of CRP, IL-1β, ET-1 and IFN-γ were increased significantly, while the levels of SV and LVEF were decreased significantly (P<0.05 or P<0.01). Compared with model group, above symptoms of rats in each administration group were improved to different extents; the levels of LVEDV and LVESV as well as the levels of CRP, IL-1β, ET-1 and IFN-γ in positive group, the levels of LVEDV in ZTY water extract medium-dose and high-dose groups, the levels of LVESV and CRP, IL-1β, ET-1, IFN-γ in ZTY water extract groups were decreased significantly. The levels of SV and LVEF in positive group, the levels of SV in ZTY water extract groups as well as the levels of LVEF in ZTY water extract medium-dose and high-dose groups were increased significantly (P<0.05 or P<0.01). CONCLUSIONS: ZTY water extract could reduce the level of ET-1 and improve blood vessels diastolic and systolic function in CHD model rats, then restore blood supply of myocardial tissue and strengthen cardiac function, by acting on inflammation cycle.
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Objective To isolate the glioblastoma multiforme stem cells (GBM-SCs) from GBM specimens and to investigate the biological role ofmiR-153 in GBM-SCs so as to explore the application of gene therapy of GBMs.Methods CD133+ cells were separated using magnetic cell sorting technique (MACS) after primary culture.Immunofluorescence staining was employed to detect the CD133,nestin,glial fibrillary acidic protein (GFAP),microtubule-associated protein 2 (MAP2) expressions; real time-PCR was used to analyze miR-153 mRNA expression in CD 133-cells and CD 133+ cells.Lipofectamine RNAiMAX was used to transfect MiR-153 mimic (miR-153 group) and scrambled control oligonucleotides (NC group) into GBM-SCs; 7 d after that,sphere formation assay was performed to determine the self-renewal ability of GBM-SCs.Real time-PCR and immunofluorescence were carried out to examine the CD133,nestin,GFAP,MAP2 mRNA and protein expressions.At last,the proliferation ability of miR-153 treated GBM-SCs and NC cells was determined by CCK-8 at 24,48,72,96 and 120 h after the transfection and the apoptosis ratio was detected by flow cytometry 3 d after the transfection.Results GBM-SCs isolated from GBM specimens could express stem cell markers CD133 and nestin; after differentiation,the cells could express astrocyte marker GFAP and neuron marker MAP2; miR-153 expression in CD133+ cells was signficantly down-regulated as compared with that in CD133-cells (P<0.05).Seven d after transfection,the number of spheres in the NC group was significantly larger than that in the miR-153 group (P<0.05); real time-PCR indicated that the mRNA expressions of CD 133 and nestin in the miR-153 group were significantly decreased,and the GFAP and MAP2 mRNA expressions were statistically increased as compared with those in the NC group (P<0.05).The results detected by immunofluorescence were in accordance with those by real time-PCR; 48,72,96h after the transfection,cell viability in cells from miR-153 group was statistically significant lower than that in the NC group (P<0.05); flow cytometry showed that the apoptosis rate of cells from the miR-153 group (9.4 1%±1.98%) was significantlyhigher than that in the NC group (4.28%±0.31%,P<0.05).Conclusion Reactivation of miR-153 expression suggests novel therapeutic strategy for GBM-SCs.
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In view of the problems that conventional artificial cartilages have no bioactivity and are prone to peel off in repeated uses as a result of insufficient strength to bond with subchondral bone, we have designed and prepared a novel kind of PVA-BG composite hydrogel as bionic artificial articular cartilage/bone composite implants. The effects of processes and conditions of preparation on the mechanical properties of implant were explored. In addition, the relationships between compression strain rate, BG content, PVA hydrogels thickness and compressive tangent modulus were also explicated. We also analyzed the effects of cancellous bone aperture, BG and PVA content on the shear strength of bonding interface of artificial articular cartilage with cancellous bone. Meanwhile, the bonding interface of artificial articular cartilage and cancellous bone was characterized by scanning electron microscopy. It was revealed that the compressive modulus of composite implants was correspondingly increased with the adding of BG content and the augments of PVA hydrogel thickness. The compressive modulus and bonding interface were both related to the apertures of cancellous bone. The compressive modulus of composite implants was 1.6-2.23 MPa and the shear strength of bonding interface was 0.63-1.21 MPa. These results demonstrated that the connection between artificial articular cartilage and cancellous bone was adequately firm.