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Newcastle disease virus-like particles (NDV VLPs) are composed of matrix protein (M) as the skeleton,with the insertion of hemagglutinin-neuraminidase and/or fusion protein.NDV VLPs are reported to be immunogenic and can induce specific humoral and cellular immune responses.However,its relationship with innate immunity remains elusive.Dendritic cells (DCs) are a group of specialized antigen presenting cells,which are crucial in connecting innate immunity and adaptive immunity.In this study,NDV VLPs and murine DCs were used to investigate the connection between NDV VLPs and innate immunity.The DC maturation induced by NDV VLPs (M+ HN) was evaluated.The results showed that NDV VLPs could be effectively taken up by DC and presented to naive T cells.NDV VLPs-induced DC significantly up-regulated the expression of MHC Ⅱ and costimulatory molecules on DC surface,and subsequently promoted the secretion of proinflammatory cytokines.This experiments also showed that different assembled NDV VLPs induced significant stimulating ability in cytokine levels.In summary,NDV VLPs can induce DC maturation,which gives insights to better understanding of VLPs-mediated innate immunity and provide information in selecting preferred NDV VLPs candidate.
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Characteristic clinical manifestations of Newcastle disease include leukopenia and immunosuppression. Peripheral blood mononuclear cells (PBMCs) are the main targets of Newcastle disease virus (NDV) infection. To survey changes in proteomic expression in chicken PBMCs following NDV infection, PBMC proteins from 30 chickens were separated using two-dimensional electrophoresis (2-DE) and subjected to mass spectrometry analysis. Quantitative intensity analysis showed that the expression of 78 proteins increased more than two-fold. Thirty-five proteins exhibited consistent changes in expression and 13 were identified as unique proteins by matrix assisted laser desorption ionization-time of flight mass spectrometer/mass spectrometer including three that were down-regulated and 10 that were up-regulated. These proteins were sorted into five groups based on function: macromolecular biosynthesis, cytoskeleton organization, metabolism, stress responses, and signal transduction. Furthermore, Western blot analysis confirmed the down-regulation of integrin-linked kinase expression and up-regulation of lamin A production. These data provide insight into the in vivo response of target cells to NDV infection at the molecular level. Additionally, results from this study have helped elucidate the molecular pathogenesis of NDV and may facilitate the development of new antiviral therapies as well as innovative diagnostic methods.
Assuntos
Animais , Proteínas Aviárias/genética , Galinhas , Regulação da Expressão Gênica , Leucócitos Mononucleares/enzimologia , Doença de Newcastle/genética , Vírus da Doença de Newcastle/fisiologia , Doenças das Aves Domésticas/genética , Proteoma , Organismos Livres de Patógenos Específicos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/veterinária , Espectrometria de Massas em Tandem/veterináriaRESUMO
A plasmids of continuous expressing shRNAs targeting the NDV NA-1 Phosphoprotein (P) gene was designed.Virus titration,Real Time RT-PCR,CPE indicated that P-specific siRNA could inhibit virus replication at 36 h post-virus infection.In future studies,a combination of siRNAs targeting the NP and L gene may be used as a tool to study NDV replication and antiviral therapy.
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To investigate whether insulin enhances canine myocardial GLUT4 translocation and glucose uptake during ischemia in vivo, physiological parameters of myocardial metabolism were measured with an autoanalyzer, and GLUT4 distribution was assessed by immunoblot analysis and immunofluorescence. The results showed insulin significantly increased ischemic myocardial sarcolemmal GLUT4 content, and it was associated with up regulation of myocardial glucose uptake. Insulin increased coronary arterio venous difference by 4 fold in ischemic region. It suggested that insulin enhances ischemic myocardial GLUT4 translocation and glucose uptake. Insulin might contribute to increase myocardial glucose uptake and utilization during ischemia.
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Objective: To investigate the relationship between serum HGF levels and clinical severity of essential hypertension (EH). Methods: The serum HGF concentrations of 44 patients with EH were measur ed by ELISA. Results: The serum HGF levels in patients with EH w ere higher than that in control. Furthermore, the serum HGF levels of EH patient s with coronary atherosclerosis (CAS) were significantly higher than those of EH patients without CAS ［(920.8±250.0） pg/ml vs （747.9±132.1） pg/ml, P <0.01］ or control ［（643.8±98.2） pg/ml, P<0.01)］.The changes of HGF l evel were correlated with the clinical courses (r=0.63, P<0.01) and stag es (r=0.69, P<0.01) of hypertension. Conclusion: HGF may be considered as a new index for the severity of hypertension and an useful bio chemical parameter for estimating the development of atherosclerosis.
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Objective: To investigate whether insulin stimulates the translocation of glucose transporter-4 (GLUT4) and glucose uptak e in ischemic myocardium. Methods: Plasma concentration of gluc ose, lactate, free fatty acid and insulin were determined by autoanalyser, and G LUT4 was studied by Western blotting analysis. Results: Insulin increased GLUT4 significantly in sarcolemma of ischemic myocardium ［(25±4)% vs (40±6)%］, and GLUT4 content in intracellular membrane decreased proporti onally. The glucose uptake increased significantly in insulin-ischemic myocardi um. The uptake of insulin-ischemic myocardium was almost 2 times that of ischem ic myocardium. Conclusion: Insulin stimulation results in GLUT4 translocation and increases glucose uptake in ischemic myocardium. When myocardi al ischemia occurs, insulin is helpful in increasing myocardial glucose uptake a nd utilization.
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Objective: To observe the effect of L-carniti ne (L-CN) in the treatment of congestive heart failure (CHF). Meth ods: Fifty-six cases of chronic CHF randomly received routine treatment (Digitalis, diuretics, vasodilator, ACEI or βblocker) or L-CN (3.0 g/d ,V D×10 d) with routine therapy. Results: The treatment efficiency of L-CN group and control group were 89.3% and 60.7% (P<0.01), respect ively. No adverse reactions related to the drug were observed. Conclusio n: L-CN with routine therapy might be a safe way to the treat CHF.
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Objective: To investigate the changes of myo cardial contractile function during myocardial stunning in calcium overload rats and the protective effects of tetrandrine. Methods: Forty-six rats were randomized into control, myocardial ischemia, myocardial stunning, low and high dose of tetrandrine groups. Another 10 rats were used to identify the calcium overload. vitamin D3 (0.3 million Unit/kg) and nicotinic acid were adm inistered. After 16 d when calcium overload occured, left anterior descending ar tery was ligated. Twenty minutes of myocardial ischemia followed by 60 min of re perfusion was induced. The contractile function parameters were determined dynam ically. At the end of experiment, myocardial cytosolic ［Ca2+］i was deter mined in various groups. In tetrandrine groups, tetrandrine (62.2 or 93.6 μmol/ kg ) was administered by gastrogavage daily.After 16 d, the rats undergone the e xperiments mentioned above. Results: Sixteen days after vitamin D3 , nicotinic acid were given, ［Ca2+］i increased by 2.6 folds (146.8±10.8 ) vs (368.5±22.6) nmol/L, (P<0.01). Whereas, ［Ca2+］i in tetrand rine groups were (210.8±16.4) and (198.6±15.3) nmol/L, which were significantl y lower than that of calcium overload group. Twenty minutes of myocardial ische mia resulted in the decrease of dp/dtmax and Vmax in all groups with the most si gnificant in stunning and calcium overload groups. The contractile function rest ored gradually after reperfusion. At all time points, dp/dtmax and Vmax in both tetrandrine groups were higher than those in both stunning and calcium overload groups. And effect with higher dose of tetrandrine were more significant than in low dose of tetrandrine. After 60 min of reperfusion, dp/dtmax in stunning, cal cium overload, low and high dose of tetrandrine groups were 49.7%, 51.5%, 71.0% and 83.4% of that in control, respectively， and Vmax were 55.0%, 49.8%, 73.9% and 77.5% of that in control, respectively. Conclusion: T he myocardial contractile function in vitamin D3-induced calcium overload gro up is impaired. On basis of myocardiocyte calcium overload, transient ischemia l eads to myocardial stunning. At the stage of ischemia, the impaired degree of my ocardial contractile function is similar to that in stunning group, suggesting a t this stage the effect of ischemia on myocardial function is greater than that of calcium overload. Tetrandrine chronically improves the myocardial function in Vitamin D3-induced calcium overload rats.
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Objective: To investigate whether there is additi ve effects of hyperinsulinemia and ischemia on expression of canine myocardial G LUT4 gene in vivo. Methods: The expression of myocardial GLU T4 was determined by semiquantitative immunoblotting.The expression of GLUT4 mRN A was determined by semiquantitative Northern blotting. Results: Dramatic changes were seen in GLUT4 mRNA and GLUT4 expression in the ischemic hearts.After infusing insulin for 8 h,regional GLUT4 mRNA and GLUT4 levels in is chemic hearts were 2.5, 2.3-fold that of expression in normal hearts(P<0.01 ). Myocardial glucose uptake in ischemic hearts was increased by 4-fold when co mpared with normal hearts(P<0.01). Conclusion: There are not only additive effects of hyperinsulinemia and low-flow ischemia on canine myoc ardial GLUT4 mRNA and GLUT4 expression in vivo, but also increase of myocar dial glucose uptake. Enhanced GLUT4 expression may be an important protective m echanism by which myocardial cells enhance glucose uptake and metabolism during low-flow ischemia.
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AIM: To investigate the mechanism underlying insulin-stimulated increase in glucose uptake during low-flow myocardial ischemia. METHODS: The expression of myocardial GLUT1 polypeptide was determined by semiquantitative immunoblotting. The expression of GLUT1 mRNA was determined by semiquantitative Northern blotting. RESULTS: After infusing insulin during low- flow myocardial ischemia for 8 h，the expression of both GLUT1 mRNA and GLUT1 polypeptide was significantly higher in experimental myocardium than that in normal myocardium. The glucose uptake was upregulated at the same time in the exprimental myocardium. CONCLUSION: Insulin enhances the expression of GLUT1 mRNA and GLUT1 polypeptide in ischemic myocardial regions. GLUT1 expression may be an important mechanism by which myocardial cells enhance glucose uptake and metabolism during low-flow myocardial ischemia.
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Objective To investigate the mechanism for that insulin facilitates increase of glucose uptake. Methods The expression of myocardial GLUT4 polypeptide was determined by semiquantitative immunoblotting. The expression of GLUT4 mRNA was determined by semiquantitative Northern blotting. Results After infusing insulin for 8 hours，the expression of GLUT4 mRNA and GLUT4 polypeptide was significantly higher in canine myocardium than in those found normal ones. The glucose uptake was upregulated at the same time．Conclusions Our findings suggest that insulin facilitates the expression of GLUT4 mRNA and GLUT4 polypeptide in canine hearts. Enhanced GLUT4 expression is one of the important molecular mechanism by which myocardial cells enhance glucose uptake by insulin stimulation.
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AIM: To investigate the mechanism underlying insulin-stimulated increase in glucose uptake during low-flow myocardial ischemia. METHODS: The expression of myocardial GLUT1 polypeptide was determined by semiquantitative immunoblotting. The expression of GLUT1 mRNA was determined by semiquantitative Northern blotting. RESULTS: After infusing insulin during low- flow myocardial ischemia for 8 h,the expression of both GLUT1 mRNA and GLUT1 polypeptide was significantly higher in experimental myocardium than that in normal myocardium. The glucose uptake was upregulated at the same time in the exprimental myocardium. CONCLUSION: Insulin enhances the expression of GLUT1 mRNA and GLUT1 polypeptide in ischemic myocardial regions. GLUT1 expression may be an important mechanism by which myocardial cells enhance glucose uptake and metabolism during low-flow myocardial ischemia. [
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The changes of Na+ -pump., and Na+, K+ and Mg2+ ions within erythrocytes were studied during treating supraventricular tachyrhythmia (SVT) by Verapamil (VR). The levels of Na+, K+ and Mg2+ within erythrocytes of 30 normal people were 77.9?2.1, 172.5?3.8 and 16.4? 0.4?mol/g Hb respectively and the level of Na+- pump of the membrane of erythrocytes was 43.9? 6.1 ?mol Pi/g Hb/h. The average levels of Na+, K+ and Mg2+ within erythrocytes of 53 SVT were 83.715.0, 143.2?12. 5, 12.6?0.5 umol/g Hb and the level of Na+-pump was 65.115.2 ?Pi/g Hb/h. After VR treatment, the level of Na+ within erythrocytes was 'obviously higher than that of normal person (P
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To determine the ATPase activity of erythrocytic membrane and intraerythrocyticionic concentrations in 20 patients with chronic renal failure (CRF). Methods: Erythrocyte membrane AT-Pase activites were determined as described by Sha1ev, and erythrocyte Na+ and Ca2+ concentrations weredetected by atomic absorption spectrophotometry. Results: The Na+-K+-ATPase, Ca2 t -ATPase, Mg2+ATPase activities were significantly lower in CRF patients than in normal individuals (P
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Objective: To observe the effect of L carnitine ( L CN) in the treatment of congestive heart failure (CHF). Methods: Fifty six cases of chronic CHF randomly received routine treatment (Digitalis, diuretics, vasodilator, ACEI or ?blocker) or L CN (3.0 g/d ,VD?10 d) with routine therapy. Results: The treatment efficiency of L CN group and control group were 89.3% and 60.7% ( P
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Objective:To investigate the changes of plasma free carnitine (FC) level in type 2 diabetes mellitus patients with hypertension and the influence of L-carnitine(L-CN) on plasma FC,blood glucose and blood viscosity. Methods:Plasma FC, blood glucose and blood viscosity were measured in 20 type 2 diabetic patients with hypertension (group 1) and 20 patients with essential hypertension (group 2) before and after they received 3. 0 g/d /,-CN intravenous infusion for ]0 d. Re-sultstlt was observed that plasma FC level was lower in group 1 [(50. 59?13. 41) ?mol/L] than in group 2 [(63. 32? 15. 23) ?mol/L,P
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Objective:To investigate the effects of chitosan on experimental rabbit atherosclerosis(AS).Methods:The models of rabbit AS were produced by a high fat diet.The rabbits were randomly assigned into normal diet(ND) group,high fat diet(HFD) group,chitosan and high fat diet(CHFD) group,chitosan and normal diet(CND) group.Fasting blood samples were collected for serum lipid assay on 0,4,8 weeks;the animals were killed on the 8th week to make aorta samples.AS plaque areas were estimated in the descending aorta dyed with oil red O and the ascending aorta dyed with H E.Results:TG,TC,LDL C in the CHFD group were lower than those in HFD group on 4 and 8 weeks;serum lipid concentration in ND group was similar with that in CND group;plaque areas in CHFD group were smaller than those in HFD group;intima thickness in CHFD group was thinner than that in HFD group.Conclusion:Chitosan reduces AS degree in rabbits of HFD group,which might be related to the decrease of serum lipid.
