CD4~+T淋巴细胞Kv1.3通道在大鼠动脉粥样硬化中的作用

目的:探讨大鼠动脉粥样硬化(AS)模型中CD4+T淋巴细胞电压门控钾通道(Kv)Kv1.3的表达、功能及其在AS中的作用。方法:采用高脂饮食法建立大鼠动脉粥样硬化模型,流式细胞术分析淋巴细胞的比例,采用免疫磁珠法分离CD4+T淋巴细胞,研究脾组织CD4+T淋巴细胞Kv1.3mRNA表达、细胞内钙离子浓度及细胞因子分泌的变化。结果:(1)AS组脾组织CD4+T淋巴细胞占总T淋巴细胞比例较对照组明显升高(74.93%±2.15%vs67.80%±2.54%,P<0.05)。(2)经刀豆蛋白A(ConA)刺激,AS组T淋巴细胞增殖程度明显高于对照组(1.1321±0.1750vs0.7971±0.0955,P<0.05)。(3)AS组脾组织CD4+T淋巴细胞在ConA刺激状态下胞内钙离子浓度明显高于对照组(H=82,P<0.05)。(4)AS组脾组织CD4+T淋巴细胞在刺激48h后较刺激24h后细胞因子(IL-2,TNF-α)分泌显著增加。(5)AS组脾组织CD4+T淋巴细胞Kv1.3mRNA表达明显高于对照组(3.670±1.579vs1)。结论:AS组脾组织CD4+T淋巴细胞比例高于对照组,CD4+T淋巴细胞Kv1.3mRNA表达增多,提示高表达Kv1.3的CD4+T淋巴细胞可能在AS的发生发展中发挥重要的作用

Role of Kv1. 3 channel of CD4--+ T lymphocytes in the formation of atherosclerosis in rat spleen

目的:探讨大鼠动脉粥样硬化(AS)模型中Cd4+T淋巴细胞电压门控钾通道(kV)kV1.3的表达、功能及其在AS中的作用。方法:采用高脂饮食法建立大鼠动脉粥样硬化模型,流式细胞术分析淋巴细胞的比例,采用免疫磁珠法分离Cd4+T淋巴细胞,研究脾组织Cd4+T淋巴细胞kV1.3MrnA表达、细胞内钙离子浓度及细胞因子分泌的变化。结果:(1)AS组脾组织Cd4+T淋巴细胞占总T淋巴细胞比例较对照组明显升高(74.93%±2.15%VS67.80%±2.54%,P<0.05)。(2)经刀豆蛋白A(COnA)刺激,AS组T淋巴细胞增殖程度明显高于对照组(1.1321±0.1750VS0.7971±0.0955,P<0.05)。(3)AS组脾组织Cd4+T淋巴细胞在COnA刺激状态下胞内钙离子浓度明显高于对照组(H=82,P<0.05)。(4)AS组脾组织Cd4+T淋巴细胞在刺激48H后较刺激24H后细胞因子(Il-2,Tnf-α)分泌显著增加。(5)AS组脾组织Cd4+T淋巴细胞kV1.3MrnA表达明显高于对照组(3.670±1.579VS1)。结论:AS组脾组织Cd4+T淋巴细胞比例高于对照组,Cd4+T淋巴细胞kV1.3MrnA表达增多,提示高表达kV1.3的Cd4+T淋巴细胞可能在AS的发生发展中发挥重要的作用。AIM: To investigate the function of voltage-gated potassium channel Kv1.3 and its possible role in CD4 + T lymphocytes in the formation of atherosclerosis ( AS) in rat spleen.METHODS: The rat atherosclerosis model was established by feeding high-fat diet.The proportion of lymphocytes was determined by flow cytometry.The CD4 + T lymphocytes were separated using immunomagnetic bead.The mRNA expression of Kv1.3 in CD4 + T lymphocytes was detected.The concentrations of intracellular calcium and cytokines were also measured.RESULTS: ( 1) The proportion of CD4 + T lymphocytes in AS group was significantly higher than that in control group ( 74.93% ± 2.15% vs 67.80% ± 2.54% ,P < 0.05) .( 2) After stimulated with concanavalin A ( ConA) ,the proliferation of CD4 + T lymphocytes in AS group was significantly higher than that in control group ( 1.1321 ± 0.1750 vs 0.7971 ± 0.0955,P < 0.05) .( 3) After stimulated with ConA,the concentration of intracellular calcium in AS group was higher than that in control group.( 4) In AS group,the releases of cytokines of IL-2 and TNF-α in AS group were significantly higher when stimulated with ConA for 48 h than that for 24 h.( 5) The mRNA expression of Kv1.3 in CD4 + T lymphocytes was greatly higher in AS group than that in control group ( 3.670 ± 1.579 vs 1) .CONCLUSION: In AS rats,the increase in CD4 + T lymphocytes as well as the augmentation of Kv1.3 mRNA expression in the cells suggest that up-regulation of Kv1.3 mRNA expression in CD4 + T lymphocytes may be involved in the mechanism of atherosclerotic formation in rat spleen.国家自然科学基金资助项目(No30871045

Ionic mechanism underlying distinctive excitability in atrium and ventricle of the heart

心肌细胞兴奋性是维持正常的心脏活动的一个重要生理因素。本研究旨在使用全细胞膜片技术探讨豚鼠心房和心室肌细胞不同兴奋性的离子机理。结果显示,心室肌细胞兴奋性比心房肌细胞低。虽然豚鼠心室肌细胞的电压门控快nA+电流(InA)密度较低,但与其兴奋性较低并不相关,因为其在阀电位附近的可用度比率比心房肌细胞高。经典内向整流钾电流(Ik1)在心室肌细胞比在心房肌细胞更大,这可能是心室肌细胞兴奋性较低的部分原因。此外,去极化引起的有内向整流特性的瞬时外向钾电流(ITOIr)在心室肌细胞较大,并可能是其兴奋性较低的主要原因。在心室肌细胞,5μMOl/l bA2+显著抑制ITOIr,增强细胞兴奋性,并使InA激活的阈电位更负,其作用独立于对InA的影响。本研究结果证明,除经典的Ik1外,ITOIr在豚鼠心房肌和心室肌细胞的兴奋性差异形成和心肌兴奋性维持中起着主要作用。然而,ITOIr增加是否会通过降低兴奋性以保护心脏,还需要进一步研究。Cellular excitability is an important physiological factor in maintaining normal cardiac activity.The present study was designed to investigate the ionic mechanism underlying different excitability in atrial and ventricular myocytes of guinea pig heart using a whole-cell patch configuration.We found that excitability is lower in ventricular myocytes than that in atrial myocytes.Although the density of voltage-gated fast Na+ current(INa) was lower in ventricular myocytes, it would not correlate to the lower excitability since its availability was greater than that in atrial myocytes around threshold potential.Classical inward rectifier K+ current(IK1) was greater in ventricular myocytes than that in atrial myocytes, which might contribute in part to the lower excitability.In addition, the transient outward K+ current with inward rectification(Itoir) elicited by depolarization was greater in ventricular myocytes than that in atrial myocytes and might contribute to the lower excitability.In ventricular myocytes, Ba2+ at 5 μmol/L significantly inhibited Itoir, enhanced excitability, and shifted the threshold potential of INa activation to more negative, and the effect was independent of affecting INa.Our results demonstrate the novel information that in addition to classical IK1, Itoir plays a major role in determining the distinctive excitability in guinea pig atrial and ventricular myocytes and maintaining cardiac excitability.More effort is required to investigate whether increase of Itoir would be protective via reducing excitability.supportedbySunChiehYehHeartFoundationofHongKongandaGeneralResearchFund(HKU771712M)fromResearchGrantCouncilofHongKon

轮状病毒VP4~*高聚体的制备及其免疫保护性评价

在前期工作中发现,截短的轮状病毒VP4~*蛋白(aa26–476)在大肠杆菌中能够以可溶形式表达,且在小鼠模型中具有较高的免疫原性和免疫保护性。本研究通过颗粒化进一步提高VP4~*蛋白的免疫保护性。通过37℃水浴加热处理24h使VP4~*蛋白多聚化,通过高效液相色谱、透射电镜、分析超离等分析VP4~*蛋白颗粒化程度,通过酶联免疫吸附试验分析颗粒化对VP4~*蛋白与中和抗体反应性的影响;通过差示量热法分析VP4~*高聚体的热稳定性;最后,通过小鼠母传抗体模型研究颗粒化对VP4~*免疫原性和免疫保护性的影响。结果表明,VP4~*蛋白高聚体结构均一,并且相比三聚体,具有更高热稳定性和中和抗体结合活性;在内毒素<20 EU/mg的条件下,与铝佐剂混合,刺激小鼠产生更高滴度的中和抗体;对轮状病毒导致的腹泻具有更高的免疫保护性。综上所述,VP4~*高聚体的研究为轮状病毒基因工程亚单位疫苗的研制提供了更广阔的思路。国家自然科学基金(No.81501741);;福建省科学技术创新平台(No.2014Y2004)资助~

Synthesis and characterization of cellulose acetate with high degree of substitution from miscanthus

以芒草为原料,用Na OH/H_2O_2溶液体系预处理制备芒草纤维,在冰醋酸环境下,以浓硫酸为催化剂与醋酸酐酯化制备芒草醋酸纤维素。优化了预处理条件:温度、时间、次数和酯化条件:催化剂量、温度、时间、醋酸酐量,最佳条件下制备出的芒草纤维的纤维素、半纤维素和木质素的质量分数分别为75.3%、17.3%、5.1%,制备出芒草醋酸纤维素的取代度DS=2.8,特性黏度[η]=1.24 d L/g,达到美国联邦贸易委员会指南认定的三醋酸纤维素标准。利用扫描电镜(SEM)和热分析(TG、DSC)对制得样品进行表征。结果表明,可以利用Na OH/H_2O_2水溶液体系预处理芒草原料制备芒草纤维,并进一步酯化制备出高取代度的醋酸纤维素。A procedure for synthesizing cellulose acetate with high degree of substitution from miscanthus biomass is developed. The miscanthus fiber is prepared by pretreatment of miscanthus biomass with Na OH / H_2O_2,which is then reacted with acetic anhydride in an acetic acid solvent to synthesize cellulose acetate by using concentrated sulfuric acid as catalyst. The effects of the pretreatment factors( such as pretreatment temperature,time and number of times) and the esterification factors( such as catalyst volume,reaction temperature,reaction time and acetic anhydride volume) are studied. Under the optimal conditions,the contents of cellulose,hemicellulose and lignin for prepared miscanthus fiber are75. 3%,17. 3% and 5. 1%,respectively. The degree of substitution( DS) and the intrinsic viscosity( [η ]) of the obtained miscanthus cellulose acetate are 2. 8 and 1. 24 d L / g,respectively. The miscanthus biomass,fiber and cellulose acetate are characterized by SEM,TG and DSC. This study shows that cellulose acetate with high degree of substitution can be prepared from miscanthus biomass pretreated with Na OH / H_2O_2.国家自然科学基金(21303142;31170067);; 福建省中青年教师教育科研项目(JA14010);; 厦门市海洋经济发展专项资金项目(14GZP59HJ29);; 福建省海洋高新产业发展专项项目(闽海洋高新[2014]25号);; 厦门大学校长基金(20720150090

支撑应力对骨小梁分布的影响及股骨头坏死因素的研究

目的通过犬股骨颈骨折螺钉内固定模型的力学检测及组织学观察,从微观角度认识骨小梁重建对股骨头坏死的影响。方法选取18只成年田园犬制作成股骨颈骨折螺钉内固定模型,于造模后12周确认所有股骨颈骨折已愈合随机分为取钉组、取钉植骨组及不取钉组,于造模后20周分离所有犬的股骨并进行股骨颈力学测试及组织学观察。结果取钉组、取钉植骨组及不取钉组断裂点载荷、最大载荷差异有统计学意义(P0.05),不取钉组与取钉植骨组断裂点载荷大于取钉组;不取钉组最大载荷大于取钉植骨组与取钉组,取钉植骨组最大载荷大于取钉组。取钉组、取钉植骨组及不取钉组骨小梁宽度与新鲜骨面积差异有统计学意义(P<0.05);不取钉组骨小梁宽度、新鲜骨面积大于取钉植骨组,且取钉植骨组大于取钉组。结论支撑应力的改变将导致骨小梁重新分布,骨小梁再分布是影响股骨头坏死塌陷的重要因素。福建省卫生系统中青年骨干人才培养项目(2014-ZQNJC-34

大鼠动脉粥样硬化斑块中T淋巴细胞亚群及Kv1.3通道蛋白的表达

目的探究Kv1.3通道蛋白与动脉粥样硬化(AS)模型中活化的T淋巴细胞间的关系。方法 Wistar雄性大鼠24只,随机分为对照组(n=10)和AS组(n=14),采用高脂饲料喂养方法建立AS模型。分别于实验开始前,实验第8周,实验第12周观察各组大鼠体重变化。于第12周处死大鼠前取血,检测血清中总胆固醇(TC)、低密度脂蛋白(LDL-L)、高密度脂蛋白(HDL-L)和甘油三酯(TG)的水平。通过病理HE染色及免疫组织化学方法观察AS斑块内T淋巴细胞亚群分布和Kv1.3通道蛋白表达的改变。结果 AS组体重、TC、LDL-C较对照组明显升高(P均<0.05);HDL-C和TG两组无差异。AS组主动脉管壁可见明显斑块形成,对照组血管壁各层的组织结构正常。AS组动脉斑块部位内膜下及中膜层可见CD4+与CD8+T淋巴细胞聚集,以CD4+T淋巴细胞聚集为主,在病变部位Kv1.3通道蛋白表达增加。对照组血管内膜、中膜中未见T淋巴细胞聚集及KV1.3通道蛋白的表达。结论 Kv1.3通道可能在调节AS斑块部T淋巴细胞亚群的激活中起着重要作用

Direct experimental evidence for detailed growth of SiO_x nanowire during CVD

在纳米线的制备中,气-液-固(VlS)生长机制得到了人们的广泛认可,但该机制的很多细节还停留在模型阶段.依托实验室自行设计的一台生长条件高度可控的高温化学气相沉积(CVd)系统,采用较为简便的方法,直接在SI片衬底上制备出了SIOX纳米线.通过严格控制实验参数,用离位观测捕捉到了纳米线的催化、形核和长大的一系列过程及其相关细节,并发现纳米线从细到粗的气-液-固(VlS)生长机制.讨论了气-液-固(VlS)机制中气态SI原子的来源以及纳米线的催化、形核和长大过程中的纳米曲率效应和“纳米熟化“现象,取得了对SIOX纳米线VlS催化生长机制的理解的突破.Among the mechanisms for nanowire growths, the vapor-liquid-solid (VLS) mechanism is the most widely accepted.Nevertheless, the growth process and relevant details for the VLS mechanism are not yet fully understood for the complicated nano processes involved.In the present article, with a precise control of temperature, gas flow, pressure, and reaction periods in a home- built high-temperature chemical vapor deposition (CVD) system, detailed processes of catalyzing, nucleation, and growth of the SiOx nanowires and a stepwise non uniformity in diameter of nano- wire were successfully traced.With analysis of these experimental results via nanocurvature and nano ripening effects, a further understanding of the vapor-liquid-solid mechanism, especially the mechanism for formation of the stepwise non uniformity in diameter of nanowires, was achieved for the first time.国家科技计划国际科技合作与交流专项(编号:2008DFA51230);国家重点基础研究发展计划(编号:2007CB936603);国家自然科学基金(批准号:60776007、90401022);中澳科技合作特别基金(编号:20050222);教育部科技重点项目(编号:105099)资

梅毒螺旋体47 ku抗原的分段表达及抗原表位分析

通过大肠杆菌表达系统表达了梅毒螺旋体47 ku膜蛋白及其N端和/或C端缺失的重组蛋白共15个,经亲和层析纯化,并通过免疫印迹和酶联免疫实验检测这组蛋白与梅毒病人血清的反应性.结果发现所有包含D结构域的重组蛋白的免疫反应性显著高于不含D结构域的蛋白,提示D结构域中可能包含47 ku膜蛋白上的一个免疫优势表位

Expression profile of voltage-gated potassium channel Kv1.3 in aorta of atherosclerosis rats

目的探讨动脉粥样硬化(AS)大鼠主动脉病变局部离子通道kV1.3的表达水平及作用。方法雄性WISTAr大鼠16只,随机分为两组:正常组(8只,予普通饮食+0.9%氯化钠溶液)和AS组(8只,予高脂饮食+维生素d3负荷)。采用组织病理学检查,观察主动脉粥样硬化病变。采用实时定量反转录-聚合酶链反应(rT-PCr)和WESTErn印迹法检测主动脉病变局部kV1.3和白细胞介素(Il)-2、干扰素(Ifn)-γ的表达水平。结果组织病理学检查证实纤维增生性AS斑块。AS组的kV1.3MrnA为(31.48±8.64)x10-3,显著高于正常组的(3.28±0.79)x10-3(P=0.012)。AS组的kV1.3、Il-2、Ifn-γ蛋白表达量分别为0.691±0.067、0.611±0.055、0.759±0.050,均显著高于正常组的0.490±0.052、0.299±0.058、0.273±0.052(P值均<0.01)。结论 kV1.3在主动脉粥样硬化病变局部的表达增高。kV1.3可能在AS的发生和发展过程中发挥着重要的作用。Objective To investigate the expression of voltage-gated potassium channel Kv1.3 in the aorta of a rat model of atherosclerosis and its role in the progress of atherosclerotic plaque formation.Methods A total of 16 male Wistar rats were randomly divided into normal control(normal diet and saline)and atherosclerosis group(high lipid diet+Vitamin D3 overload),with 8 rats in each group.In 16 weeks later,all rats were killed after weighing,and their blood samples and aorta were collected.Pathological changes of the rat aortic artery were observed with HE staining.Real time RT-PCR and Western blotting analysis were used to determine the mRNA and protein expression of Kv1.3 and the protein expressions of interleukin(IL)-2 and interfereron(IFN)-γ.Results Pathological changes showed that fiber proliferative atherosclerotic plaques were found in the aorta of atherosclerosis group,with inflammatory cells infiltrating in the local lesion.Real time RT-PCR analysis showed that the expression of Kv1.3 mRNA in the aorta was increased significantly in the atherosclerosis rats than that in the controls([31.48±8.64]×10-3 vs.[3.28±0.79]×10-3,P<0.05).Western blotting analysis showed that the protein expression of Kv1.3,IL-2 and IFN-γ in the aorta were also increased significantly in the atherosclerotic rats than that in the controls(Kv1.3[0.691±0.067] vs.[0.490±0.052],IL-2 [0.611±0.055] vs.[0.299±0.058],IFN-γ [0.759±0.050] vs.[0.273±0.052],P<0.01 n=8).Conclusion The expression of Kv1.3 potassium channels is increased in the plaques of atherosclerotic rats.Kv1.3 may play an important role in the development and progression of atherosclerosis.国家自然科学基金资助项目(30871045