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Objective To study the effect of a Chinese medicine, Huanglian Jiedu Decoction ( HLJDD) , on ath-erosclerotic plaque, inflammatory factors and regulatory T cells in ApoE-/- mice. Methods High fat diet was given to ApoE-/- mice to establish an atherosclerosis model. 40 male ApoE-/- mice were randomly divided into five groups:model group, simvastatin group, low, moderate and high dose HLJDD groups (n =8). HLJDD was intragastrically administered in a dose of 3. 5, 7. 0, or 14. 0 g/( kg?bw) once dailg for 16 weeks. The dose of simvastatin was 5. 0 g/( kg?bw). An-other 8 male C57BL/6J mice were taken as control group. At the end of the 29-week experiment, all of the mice were sacri-ficed. The aortic plaques, level of blood lipids, inflammatory factors, Tregs number and the level of Foxp3 mRNA were de-tected and analyzed by ELISA, flow cytometry and RT-PCR, respectively. Results Compared with the control group, the aortic plaques were much larger in the model group, and the levels of TC, TG, LDL-C, IL-6, hs-CRP, and TNF-α were significantly higher than those in the control group (P<0. 01 for all). Meanwhile, the levels of HDL-C, IL-10, TGF-βand Foxp3 mRNA were much lower than those in the control group ( P<0. 01 for all) , and the Tregs numbers were less than that in the control group (P<0. 01). HLJDD regulated the blood lipids in ApoE-/ - mice and decreased the levels of IL-6, hs-CRP, andTNF-α, however increased the levels of IL-10, TGF-β and Foxp3 mRNA. At the same time, it in-creased Tregs number in the ApoE-/ - mice. Compared with the model group, the difference was statistically significancet (P<0. 01). Conclusions HLJDD can significantly alleviate the aortic plaque damages in ApoE-/ - mice. It may be re-lated to the up-regulation of Tregs, which can lead to decrease the expression of serum pro-inflammatory factors such like IL-10, hs-CRP and TNF-α.
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Object To study the effect of puerarin on the expression of inflammatory factors and miR-155-3p in human umbilical vein endothelial cells ( HUVEC) induced by visfatin.Methods The HUVEC cell injury model was es-tablished with visfatin.Cell proliferation was measured by MTT assay.Cell apoptosis was detected by flow cytometry.The level of CRP and NF-κB was detected by ELISA, and the expression of miR-155-3p was detected by RT-PCR.The expres-sion of myeloid differentiation factor 88 ( MyD88) was identified by western blotting.Results Visfatin induced cell prolif-eration and inhibited apoptosis in HUVEC, meanwhile the expressions of both CRP and NF-κB were significantly increased, compared with that of the control group (P<0.01).Puerarin at moderate and high concentrations obviously reduced the HUVEC injury induced by visfatin, mainly through down-regulating the expression of CRP and NF-κB, as well as up-regu-lating the level of miR-155-3p in the HUVEC.MiR-155-3p mimic markedly decreased the level of MyD88, CRP and NF-κB in the HUVEC induced by visfatin (P<0.05).Conclusions Pueprarin obviously alleviates HUVEC injury induced by visfatin, probably related to down-regulating the level of MyD88, CRP, NF-κB, and up-regulating the expression of miR-155-3p in HUVEC.
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BACKGROUND: siRNA transfection is a key step in RNA interference. The methods of cardiomyocytes transfection were most use of plasmids as vector to transfect long-chain shRNA. However, the processes were complex. It was a simple efficient and low-toxic method that liposomes-mediatad chernosynthesis siRNA transfection. It was useful for expanding RNA interference application.OBJECTIVE: To choice the optimal concentration of Iiposomes-mediated chemosynthesis siRNA transfection, and to discover a simple efficient RNA interference application.METHODS: CY3-Negative siRNA was mediated by lipid-besed agent siPORT~(TM) NeoFX~(TM) to transfect cardiomyocytes. A blank control group was set. After 24 hours, the transfection efficiency and apoptotic rate were evaluated by fluorescent microscope and flow cytometer to select an optimal concentration. Based the best concentration, siRNA PHB was transfected to cerdiomyocytes. 48 hours later, the expression of PHB was tested.RESULTS AND CONCLUSION: With the increased concentration of CY3-Negative siRNA, the number of cells emitted red fluorescence grew under fluorescence microscope, and the transfection efficiency was also increased (P＜0.05). The best concentration was 30 nmol/L (P＜0.05). There was no significant difference in apoptotic rate between transfected groups and the control group (P ＞ 0.05). The PHB expression of cardiomyocytes transfected siRNA PHB was dropped by 74.11% on average (P＜0.05). The results indicated that lipid based agent siPORT~(TM) NeoFX~(TM) was suitable to transfect chemosynthesis siRNA to cardiomyocytes, and the best transfection concentration of siRNA was 30 nmol/L.
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Objective To investigate the anti-apoptotic mechanism of tanshinone ⅡA and the function of prohibitin(PHB)onmyocardial cells apoptosis induced by hydrogen peroxide(H2O2).Methods Myocardial cells were primary culturedneonate rat were cultured in medium with 200 μmol/L H2O2,and the medium was supplemented with tanshinone ⅡA(1 × 10-4 mol/L)in advance for 24 h.PHB in myocardial cells was knocked down by RNA interference,and theexpression level of PHB was determined by Western blotting analysis.Flow cytometric analysis was used to detectapoptosis rate,intracellular calcium concentration([Ca2+]i),and mitochondrial membrane potential(MMP).ResultsH2O2-mediated cell apoptosis resulted in activation of PHB,increasing of[Ca2+]i,and decreasing of MMP.TanshinoneⅡA profoundly inhibited myocardial cell apoptosis induced by H2O2,decreased[Ca2+]i,and increased MMP.Specificsilence of PHB by siRNA down-regulated the expression level of PHB,increased apoptosis rate and[Ca2+]i,and decreasedMMP.Conclusion The results demonstrate that tanshinone ⅡA could attenuate apoptosis induced by H2O2,and theactivation of PHB induced by H2O2 is the major regulatory pathway of cyto-protective gene expression against oxidativestress.
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Objective To anlayze the imaging diagnosis of spinal primary tumor.Methods Fifty and nine cases of spinal primary tumors confirmed by pathology were analyzed retrospectively.Results Of 59 cases,there were chordoma in 23,hemangioma in 7,giant cell tumor of bone in 11,aneurysmal bone cyst in 5,osteoblastoma in 7,osteogenous sarcoma in 2,myeloma in 2,spinal eosinophilic granuloma in 1 and primary bone lymphoma in 1.These tumors had its own imaging characteristics.Conclusion Imaging examinations are of significant value for the clinical treatment of spinal primary bone tumor.
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Objective To observe the effect of tanshinoneⅡA (TⅡA) on arrhythmia and levels of ischemia modified albumin (IMA) and heart type-fatty acid binding protein (H-FABP) in rats with myocardial ischemia-reperfusion injury.Methods Forty Wistar rats were equally randomized into 5 groups:pseudo-operation group (normal saline),model group (normal saline),and high-,middle-and low-dose TⅡA groups (in the dose of 20,10 and 5 mg?kg-1?d-1 respectively).The rats were treated for 7 days,and received operation 2 hours after the last medication.Rats in pseudo-operation group received threading without ligation,while animal model of myocardial ischemia-reperfusion injury was established by ligating the left-anterior coronary artery for 15 minutes and then loosening for 30 minutes.During the operation,electrocardiogram was traced continuously to observe the arrhythmia.HE staining was used to observe the pathological changes of myocardium.Levels of IMA and H-FABP in myocardium were determined by enzyme-linked immunosorbent assay (ELISA).Results Ischemic changes of the myocardium and arrhythmia occurred,myocardial IMA level increased and H-FABP level decreased in the model rats.TanshinoneⅡA obviously attenuated arrhythmia caused by ischemia-reperfusion injury (P
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Objective: The caracteristics of rapid arrhythmia in the south region of our country are simultaneous appearance of phlegm. heat, blood stasis and deficiency, and slow arrhythmia are intermingled deficiency. blood stasis and phlegm. Thus, they were treated respectively by clearing away heat, eliminating Yang and removing blood stasis, and warming Yang to remove blood stasis respectively. Results: The total effective rate was 96. 55% in 261 cases of rapid arrhythmia treated by modified Ding Xin Decoction, and 78. 46% in the control group with a significant difference between the two groups (P