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干细胞脑内移植有效标记研究:绿色荧光蛋白质粒标记的应用价值 被引量:15
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作者 关云谦 陈彪 +2 位作者 刘平 邹春林 张愚 《中国临床康复》 CSCD 2004年第13期2506-2508,T002,共4页
目的:观察绿色荧光蛋白(greenfluorescentprotein,GFP)质粒转化小鼠胚胎干细胞(embryonicstemcell,ESC)的效果,以及将转化的胚胎干细胞移植入正常大鼠纹状体后,GFP质粒作为细胞存活情况示踪剂的效果。方法:首先利用感受态大肠杆菌提取... 目的:观察绿色荧光蛋白(greenfluorescentprotein,GFP)质粒转化小鼠胚胎干细胞(embryonicstemcell,ESC)的效果,以及将转化的胚胎干细胞移植入正常大鼠纹状体后,GFP质粒作为细胞存活情况示踪剂的效果。方法:首先利用感受态大肠杆菌提取大量高纯度GFP质粒DNA,然后将GFP质粒与脂质体孵育形成的转化复合物和小鼠胚胎干细胞共同孵育,使GFP质粒转入胚胎干细胞;筛选表达GFP的胚胎干细胞。将筛选后表达GFP的ESC移植入活体大鼠纹状体内,移植21d后,观察表达绿色荧光蛋白的移植细胞的存活情况。结果:GFP质粒转化以后的ES细胞团和单细胞都表达亮绿色的GFP,细胞打散计数证实大约60%的细胞携带GFP,移植21d后,大鼠脑内可见大量表达GFP的移植细胞。结论:脂质体可以将GFP质粒转入鼠的胚胎干细胞,携带有GFP的ES移植后21d,GFP可以作为示踪剂观察移植细胞的存活状况。脂质体辅助的GFP质粒转化胚胎干细胞是移植示踪的较好方法。 展开更多
关键词 干细胞脑内移植 有效标记 绿色荧光蛋白质标记 胚胎干细胞 转基因技术
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Hep G2细胞中乙肝病毒衣壳包裹绿色荧光蛋白质粒表达的研究 被引量:1
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作者 潘德键 王东林 +2 位作者 王卫兵 周锡建 陈正堂 《中华肿瘤防治杂志》 CAS 2007年第16期1201-1203,共3页
目的:观察乙型肝炎病毒衣壳(hep-atitis B virus envelope,HBVE)包裹绿色荧光蛋白质粒在肝癌细胞株(Hep G2细胞)中的表达效果是否优于脂质体包裹绿色荧光蛋白质粒。方法:采用PEG8000病毒浓缩法、β-丙内脂法从Hep G2.2.15细胞上清液中制... 目的:观察乙型肝炎病毒衣壳(hep-atitis B virus envelope,HBVE)包裹绿色荧光蛋白质粒在肝癌细胞株(Hep G2细胞)中的表达效果是否优于脂质体包裹绿色荧光蛋白质粒。方法:采用PEG8000病毒浓缩法、β-丙内脂法从Hep G2.2.15细胞上清液中制备HBVE基因转移载体,分别用HBVE及脂质体包裹绿色荧光蛋白质粒(pIRS2-EGFP)后转染Hep G2细胞,通过荧光显微镜观察绿色荧光蛋白表达情况,流式细胞仪检测细胞发光率。结果:荧光显微镜下观察可见,脂质体组及HBVE组均可见绿色荧光蛋白的表达,HBVE组较之脂质体组具有更高的荧光强度;流式细胞仪检测结果显示,脂质体组的转染效率为49.97%,而HBVE组转染效率为70.65%,其转染效率及荧光强度明显高于脂质体组,P=0.000。结论:HBVE包裹绿色荧光蛋白质粒较之脂质体包裹在HepG2细胞中具有更好的转染效果。 展开更多
关键词 乙型肝炎病毒衣壳 脂质体 绿色荧光蛋白质 基因转移
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水溶性荧光聚合物纳米粒的制备及双光子成像
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作者 梁淑彩 陈小慧 +2 位作者 刘衍斌 秦蒙 鄢国平 《分析化学》 SCIE EI CAS CSCD 北大核心 2014年第5期648-653,共6页
报道了一种能用于活细胞双光子荧光( TPF)成像的水溶性聚合物纳米粒。首先以含双羧基的萘酰亚胺作为交联剂和荧光标记试剂,通过对聚乙烯亚胺发生交联反应制备纳米粒子,然后对其结构形态、单双光子荧光性能及细胞毒性进行测试。结果... 报道了一种能用于活细胞双光子荧光( TPF)成像的水溶性聚合物纳米粒。首先以含双羧基的萘酰亚胺作为交联剂和荧光标记试剂,通过对聚乙烯亚胺发生交联反应制备纳米粒子,然后对其结构形态、单双光子荧光性能及细胞毒性进行测试。结果表明,获得的纳米粒为球形,粒径为5~10 nm;以443或800 nm为激发波长,荧光发射波长均为536 nm;在pH 4.0~9.0范围内,其荧光无明显变化;在pH 7.4的溶液中和激发光为443 nm 的条件下,对其连续测定1.2万次后荧光强度变化不超过1%,说明其酸碱稳定性和光稳定性较好;浓度在15 mg/L以下及与细胞作用时间在24 h以内细胞毒性较低。最后,用双光子共聚焦荧光显微镜观察了其在 Hela 细胞中的 TPF 成像性能。将Hela细胞与纳米粒共同孵育2 h后,在800 nm激光激发下,在细胞中可观察到其绿色荧光。此纳米粒可望用于靶向性双光子荧光成像探针的开发。 展开更多
关键词 萘酰亚胺 聚乙烯亚胺 荧光聚合物纳米 双光子成像
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新型壳聚糖荧光纳米粒的制备及pH敏感性能研究 被引量:5
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作者 余慧 梁淑彩 +2 位作者 钟海迪 付婷 鄢国平 《分析化学》 SCIE EI CAS CSCD 北大核心 2011年第3期409-413,共5页
以合成的具有羧基官能团的萘酰亚胺类化合物为荧光团,通过酰氯化法活化其中的羧基,并选用生物相容性较好的水溶性高分子聚合物——羧甲基壳聚糖(CMCS)为基质材料,以化学键合的方式将荧光团引入到CMCS基体中,得到新型荧光材料萘酰亚胺修... 以合成的具有羧基官能团的萘酰亚胺类化合物为荧光团,通过酰氯化法活化其中的羧基,并选用生物相容性较好的水溶性高分子聚合物——羧甲基壳聚糖(CMCS)为基质材料,以化学键合的方式将荧光团引入到CMCS基体中,得到新型荧光材料萘酰亚胺修饰羧甲基壳聚糖CMCS3N。通过红外光谱、紫外光谱及透射电子显微镜对CMCS3N的结构和形态进行表征,证明其具有纳米结构,粒径约40 nm。荧光显微镜成像结果表明:在紫外光激发下,CMCS3N发出黄绿色荧光。荧光光谱测试该荧光纳米粒的激发波长和发射波长分别为409和534 nm,在pH 5~8范围内,其荧光对pH值敏感,且1.0×10-5mol/L常见金属离子对其荧光无明显影响。此荧光纳米粒可望作为pH敏感荧光探针,用于生物体内H+成像研究。 展开更多
关键词 萘酰亚胺 羧甲基壳聚糖 荧光纳米 PH敏感
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聚丙烯酸/萘酰亚胺纳米粒荧光增强法测定蛋白质 被引量:1
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作者 李杰鹏 梁淑彩 +2 位作者 余慧 刘衍斌 鄢国平 《分析科学学报》 CAS CSCD 北大核心 2015年第6期779-782,共4页
制备了水溶性的聚丙烯酸/萘酰亚胺荧光纳米粒,并利用牛血清白蛋白(BSA)对其荧光的显著增强效应,建立了测定蛋白质的新方法。在pH=3.0磷酸盐缓冲溶液及室温条件下,该纳米粒与BSA的作用5 min内即完成。以444nm为激发波长,在535nm处测定... 制备了水溶性的聚丙烯酸/萘酰亚胺荧光纳米粒,并利用牛血清白蛋白(BSA)对其荧光的显著增强效应,建立了测定蛋白质的新方法。在pH=3.0磷酸盐缓冲溶液及室温条件下,该纳米粒与BSA的作用5 min内即完成。以444nm为激发波长,在535nm处测定纳米粒/BSA体系的荧光,其荧光强度增加与0.15~7.5μmol·L-1浓度范围内的BSA呈线性关系,检测限为0.095μmol·L-1。该方法简单、快速、灵敏且重现性好,用于BSA合成样品及牛奶样品中蛋白质的测定,回收率范围为96.7%~102.6%,相对标准偏差(RSD)在3.91%以内。 展开更多
关键词 蛋白质 荧光纳米 聚丙烯酸 萘酰亚胺 荧光增强法
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用有色荧光塑料碎粒标志小鱼
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作者 邓思明 《现代渔业信息》 1987年第6期F003-F003,共1页
鱼类标志放流是生态学研究的重要措施之一,在以往多种标志方法中,直接放置标志牌的方法仍被广泛运用,尤其是以重氮酸基胶片做成的各种标志物,更被大家采用,因为这种标志物放置鱼体后的固着率很高,但是对小鱼来说,仍然偏大,而且... 鱼类标志放流是生态学研究的重要措施之一,在以往多种标志方法中,直接放置标志牌的方法仍被广泛运用,尤其是以重氮酸基胶片做成的各种标志物,更被大家采用,因为这种标志物放置鱼体后的固着率很高,但是对小鱼来说,仍然偏大,而且标志时很费时间,取下时还需各别鉴定。 展开更多
关键词 鱼类 标志放流 生态学研究 有色荧光塑料碎 固着率
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超声微泡造影剂携绿色荧光蛋白质粒转染大鼠股骨头的研究
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作者 陈磊 张莉 《韶关学院学报》 2019年第3期57-60,共4页
以SD大鼠为实验对象,研究超声微泡造影剂携绿色荧光蛋白质粒转染股骨头的可行性和安全性.分组实验后发现各组大鼠大体形态状况以及HE染色观察未发现骨组织损伤;应用超声微泡造影剂为载体的大鼠股骨头,绿色荧光蛋白质粒表达高于其他各组... 以SD大鼠为实验对象,研究超声微泡造影剂携绿色荧光蛋白质粒转染股骨头的可行性和安全性.分组实验后发现各组大鼠大体形态状况以及HE染色观察未发现骨组织损伤;应用超声微泡造影剂为载体的大鼠股骨头,绿色荧光蛋白质粒表达高于其他各组.故认为超声微泡造影剂携绿色荧光蛋白质粒转染大鼠股骨头组织是安全和可行的. 展开更多
关键词 超声微泡造影剂 绿色荧光蛋白质 转染 股骨头
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急性心肌梗死患者血浆游离线粒体DNA拷贝数定量分析及临床意义 被引量:5
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作者 周双艳 杜建明 +2 位作者 杜怡清 赵克斌 杨泽华 《检验医学与临床》 CAS 2017年第9期1268-1270,共3页
目的探究急性心肌梗死(AMI)后血浆游离线粒体DNA拷贝数变化及其临床意义。方法采集50例健康体检者血浆标本为对照组,50例AMI患者为AMI组,应用荧光定量PCR法测定其循环游离线粒体DNA拷贝数。结果对照组血浆游离线粒体DNA拷贝数为4×1... 目的探究急性心肌梗死(AMI)后血浆游离线粒体DNA拷贝数变化及其临床意义。方法采集50例健康体检者血浆标本为对照组,50例AMI患者为AMI组,应用荧光定量PCR法测定其循环游离线粒体DNA拷贝数。结果对照组血浆游离线粒体DNA拷贝数为4×10~4(2.5×10~4,9.5×10~4)copies/mL;AMI组血浆游离线粒体DNA拷贝数为2.2×10~5(4.9×10~5,0.8×10~5)copies/mL,差异有统计学意义(P<0.05)。结论 AMI患者血浆游离线粒体DNA拷贝数升高,血浆游离线粒体DNA拷贝数检测可能成为辅助诊断心肌梗死的生物学标志。 展开更多
关键词 急性心肌梗死 线体DNA 荧光定量PCR
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荧光标记豇豆花叶病毒的制备及细胞成像
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作者 陈小慧 梁淑彩 +2 位作者 刘衍斌 仰望 鄢国平 《分析科学学报》 CAS CSCD 北大核心 2013年第1期6-10,共5页
利用与氨基选择性反应的荧光染料5-羧基荧光素琥珀酰亚胺酯(5-FAMSE)对豇豆花叶病毒(CPMV)表面的氨基进行修饰,制备了荧光功能化CPMV纳米粒(5-FAM/CPMV)。对其结构形态和单、双光子荧光进行了测试,用毛细管电泳对其表面特性进行了考察,... 利用与氨基选择性反应的荧光染料5-羧基荧光素琥珀酰亚胺酯(5-FAMSE)对豇豆花叶病毒(CPMV)表面的氨基进行修饰,制备了荧光功能化CPMV纳米粒(5-FAM/CPMV)。对其结构形态和单、双光子荧光进行了测试,用毛细管电泳对其表面特性进行了考察,并将其用于Hela细胞的双光子荧光成像。研究表明,目标纳米粒粒径均匀,仍具有天然CPMV的特性,能进入Hela细胞,在波长800nm激光激发下可以成功地用于肿瘤细胞的成像分析,可望用于肿瘤靶向双光子荧光细胞成像方面的研究。 展开更多
关键词 豇豆花叶病毒 荧光纳米 双光子荧光成像
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饲料中黄曲霉毒素B_1快速检测方法的研究 被引量:2
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作者 齐德生 于炎湖 +1 位作者 刘耘 李玉礼 《粮食与饲料工业》 CAS 北大核心 2000年第11期49-49,共1页
取 2 0g被检样品 ,在 36 0nm紫外线下观察 ,如无亮黄绿色荧光粒 ,可基本判定饲料未受黄曲霉毒素B1污染 ;如有 1~ 4个亮黄绿色荧光粒 ,可疑为饲料黄曲霉毒素B1污染 ;如有 4个以上黄绿色荧光粒 。
关键词 饲料 黄曲霉毒素B1 快速检测 紫外线 荧光粒
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Preparation and Fluorescence Properties of Co-doped Nanocomposite Film Based on Supra Molecular Structure 被引量:3
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作者 李贵安 李貅 +4 位作者 宋建平 李发荣 马少华 张玉荣 方晓玲 《Chinese Journal of Chemical Physics》 SCIE CAS CSCD 北大核心 2006年第2期183-186,共4页
A novel materials design procedure based on the co-doping of metal nanoparticle and azo dye compound (MNPADC) is developed to improve the properties of functional molecules. The synthesized materials were characteri... A novel materials design procedure based on the co-doping of metal nanoparticle and azo dye compound (MNPADC) is developed to improve the properties of functional molecules. The synthesized materials were characterized by transmission electron micrograph (TEM), ultraviolet-visible absorption spectra (UV-Vis) and fluorescence spectra (FS). It was found that the fluorescence intensity of methyl orange (MO) was enhanced by 5 times in the aqueous composite system doped with silver nanoparticles whereas it was reduced by 15% and 20% in composite films with co-mixing and coating structures, respectively. The results indicate that the properties of functional molecules can be greatly improved in composite film with supra molecular structure and that the procedure presented here is effective. 展开更多
关键词 Supra molecular structure Nanocomposite film Silver nanoparticles Methyl orange Fluorescence properties
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SA脂质体介导报告基因转染耳蜗的实验研究 被引量:5
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作者 时 利 翟所强 +1 位作者 杨伟炎 董明敏 《中国耳鼻咽喉颅底外科杂志》 CAS 2002年第2期105-107,I002,共4页
目的 探索直接注射质粒及SA脂质体与质粒复合物在豚鼠耳蜗内表达的情况。方法 以构建的含绿色荧光蛋白基因pEGFP cDNA的重组真核表达质粒为外源基因,与SA脂质体一起转染内耳耳蜗毛细胞;通过荧光显微镜观察绿色荧光蛋白的表达。结果 使... 目的 探索直接注射质粒及SA脂质体与质粒复合物在豚鼠耳蜗内表达的情况。方法 以构建的含绿色荧光蛋白基因pEGFP cDNA的重组真核表达质粒为外源基因,与SA脂质体一起转染内耳耳蜗毛细胞;通过荧光显微镜观察绿色荧光蛋白的表达。结果 使用阳离子脂质体SA的脂质体质粒复合物组转染耳蜗,表达效率高于单纯质粒组,于转染后48h表达出的蛋白量最大。结论 应用脂质体及直接注射法是使外源基因在豚鼠耳蜗内表达的简便、有效途径。 展开更多
关键词 听力障碍/治疗 基因疗法 增强型绿色荧光蛋白质pEGFP SA脂质体 耳蜗
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阳离子脂质体介导pEGFP-N1转染HepG2.2.15细胞效率初探 被引量:2
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作者 朱晓莹 王燕菲 《右江民族医学院学报》 2012年第4期458-460,共3页
目的探讨阳离子脂质体介导绿色荧光蛋白质粒转染HepG2.2.15细胞的转染效率。方法以阳离子脂质体包裹绿色荧光蛋白质粒转染HepG2.2.15细胞,观察不同配比的脂质体与绿色荧光蛋白的转染率。结果对照组未见绿色荧光蛋白表达,各实验组HepG2.2... 目的探讨阳离子脂质体介导绿色荧光蛋白质粒转染HepG2.2.15细胞的转染效率。方法以阳离子脂质体包裹绿色荧光蛋白质粒转染HepG2.2.15细胞,观察不同配比的脂质体与绿色荧光蛋白的转染率。结果对照组未见绿色荧光蛋白表达,各实验组HepG2.2.15细胞均可见绿色荧光蛋白表达,表达量随脂质体浓度增加而增加(P<0.05),最高转染率为23.22%。结论阳离子脂质体能有效地介导绿色荧光蛋白质粒转染HepG2.2.15细胞。 展开更多
关键词 阳离子脂质体 绿色荧光蛋白质 HEPG2.2.15细胞 转染效率
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Preparation of fluorescence starch-nanoparticle and its application as plant transgenic vehicle 被引量:6
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作者 刘俊 王凤华 +4 位作者 王玲玲 肖苏尧 童春意 唐冬英 刘选眀 《Journal of Central South University of Technology》 EI 2008年第6期768-773,共6页
Starch-nanoparticles were synthesized in water-in-oil microemusion at room temperature, and the starch-nanoparticles were coated with poly-L-lysine. The surface of the starch-nanoparticles was combined with fluorescen... Starch-nanoparticles were synthesized in water-in-oil microemusion at room temperature, and the starch-nanoparticles were coated with poly-L-lysine. The surface of the starch-nanoparticles was combined with fluorescence material Ru(bpy)32+·6H2O, and then the particles were characterized via transmission electron microscope. The fluorescence nanoparticles were conjugated with plasmid DNA to form complexes, and then treated with ultrasound and DNase I. pEGAD plasmid DNA-nanoparticle complexes were co-cultured with plant suspension cells of Dioscrea Zigiberensis G H Wright, and treated with ultrasound. The results show that the diameter of the fluorescence starch-nanoparticles is 50-100 nm. DNA-nanoparticle complexes can protect DNA from ultrasound damage as well as from DNase I cleavage. Mediated by ultrasound, pEGAD plasmid DNA-nanoparticle complexes can pierce into the cell wall, cell membrane and nucleus membrane of plant suspension cells. The green fluorescence protein(GFP) gene at a high frequency exceeds 5%. This nano-biomaterial can efficiently solve the problem that exterior genes cannot traverse the plant cell wall easily. 展开更多
关键词 fluorescence nanoparticle PLANT gene vehicle ultrasound-mediation transformation
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Adaption of Au Nanoparticles and CdTe Quantum Dots in DNA Detection 被引量:4
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作者 代昭 张纪梅 +4 位作者 董全喜 郭宁 许世超 孙波 步月华 《Chinese Journal of Chemical Engineering》 SCIE EI CAS CSCD 2007年第6期791-794,共4页
A DNA fluorescence probe system based on fluorescence resonance energy transfer (FRET) from CdTe quantum dot (QD) donors to Au nanoparticle (AuNP) acceptors is presented. CdTe QDs, 2.5nm in diameter, as energy d... A DNA fluorescence probe system based on fluorescence resonance energy transfer (FRET) from CdTe quantum dot (QD) donors to Au nanoparticle (AuNP) acceptors is presented. CdTe QDs, 2.5nm in diameter, as energy donors, were prepared in water. Au nanoparticles, 16nm in diameter, as energy acceptors, were prepared from gold chloride by reduction. CdTe QDs were linked to 5'-NH2-DNA through 1-ethyl-3-(dimethylaminopropyl)car- bodiimide hydrochloride (EDC) as a linker, and the 3'-SH-DNA was self-assembled onto the surface of AuNPs. The hybridization of complementary double stranded DNA (dsDNA) bound to the QDs and AuNPs (CdTe-dsDNA-Au) determined the FRET distance of CdTe QDs and Au nanoparticles. Compared to the fluorescence of CdTe-DNA, the fluorescence of CdTe-DNA-Au conjugates decreased extremely, which indicated that the FRET occurred between CdTe QDs and Au nanoparticles. The fluorescence change of this conjugate depended on the ratio of Au-DNA to CdTe-DNA. When the AuNPs-DNA to QD-DNA ratio was 10:1, the FRET efficiency reached a maximum. The probe system would have a certain degree of fluorescence recovery when a complementary single stranded DNA was introduced into this system, which showed that the distance between CdTe QDs and Au nanoparticles was increased. 展开更多
关键词 CDTE quantum dots Au nanoparficle fluorescence resonance energy transfer DNA
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Blocking NF-kB nuclear translocation leads to p53-related autophagy activation and cell apoptosis 被引量:25
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作者 Bao-Song Zhu Chun-Gen Xing +3 位作者 Fang Lin xiao-Qing Fan Kui Zhao Zheng-Hong Qin 《World Journal of Gastroenterology》 SCIE CAS CSCD 2011年第4期478-487,共10页
AIM: To investigate the anti-tumor effects of nuclear factor-κB (NF-κB) inhibitor SN50 and related mechanisms of SGC7901 human gastric carcinoma cells. METHODS: MTT assay was used to determine the cytotoxic effects ... AIM: To investigate the anti-tumor effects of nuclear factor-κB (NF-κB) inhibitor SN50 and related mechanisms of SGC7901 human gastric carcinoma cells. METHODS: MTT assay was used to determine the cytotoxic effects of SN50 in gastric cancer cell line SGC7901. Hoechst 33258 staining was used to detect apoptosis morphological changes after SN50 treatment. Activation of autophagy was monitored with monodansylcadaverine (MDC) staining after SN50 treatment.Immunofluorescence staining was used to detect the expression of light chain 3 (LC3). Mitochondrial membrane potential was measured using the fluorescent probe JC-1. Western blotting analysis were used to determine the expression of proteins involved in apoptosis and autophagy including p53, p53 upregulated modulator of apoptosis (PUMA), damage-regulated autophagy modulator (DRAM), LC3 and Beclin 1. We detected the effects of p53-mediated autophagy activation on the apoptosis of SGC7901 cells with the p53 inhibitor pifithrin-α. RESULTS: The viability of SGC7901 cells was inhibited after SN50 treatment. Inductions in the expression of apoptotic protein p53 and PUMA as well as autophagic protein DRAM, LC3 and Beclin 1 were detected with Western blotting analysis. SN50-treated cells exhibited punctuate microtubule-associated protein 1 LC3 in immunoreactivity and MDC-labeled vesicles increased after treatment of SN50 by MDC staining. Collapse of mitochondrial membrane potential Δψ were detected for 6 to 24 h after SN50 treatment. SN50-induced increases in PUMA, DRAM, LC3 and Beclin 1 and cell death were blocked by the p53 specific inhibitor pifithrin-α. CONCLUSION: The anti-tumor activity of NF-κB inhibitors is associated with p53-mediated activation of autophagy. 展开更多
关键词 Nuclear factor-κB SN50 AUTOPHAGY P53 Cell apoptosis
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Microbubble-enhanced ultrasound exposure improves gene transfer in vascular endothelial cells 被引量:21
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作者 Fang Nie Hui-Xiong Xu +1 位作者 Qing Tang Ming-De Lu 《World Journal of Gastroenterology》 SCIE CAS CSCD 2006年第46期7508-7513,共6页
AIM: To explore the effects of ultrasound exposure combined with microbubble contrast agent (SonoVue) on the permeability of the cellular membrane and on the expression of plasrnid DNA encoding enhanced green fluor... AIM: To explore the effects of ultrasound exposure combined with microbubble contrast agent (SonoVue) on the permeability of the cellular membrane and on the expression of plasrnid DNA encoding enhanced green fluorescent protein (pEGFP) transfer into human umbilical vein endothelial cells (HUVECs). METHODS: HUVECs with fluorescein isothiocyanatedextran (FD500) and HUVECs with pEGFP were exposed to continuous wave (1.9 MHz, 80.0 mW/cm^2) for 5 min, with or without a SonoVue. The percentage of FD500 taken by the HUVECs and the transient expression rate of pEGFP in the HUVECs were examined by fluorescence microscopy and flow cytornetry, respectively. RESULTS: The percentage of FDS00-positive HUVECs in the group of ultrasound exposure combined with SonoVue was significantly higher than that of the group of ultrasound exposure alone (24.0%± 5.5% vs 66.6% ± 4.1%, P 〈 0.001). Compared with the group of ultrasound exposure alone, the transfection expression rate of pEGFP in HUVECs was markedly increased with the addition of SonoVue (16.1% ± 1.9% vs 1.5% ± 0.2%, P 〈 0.001). No statistical significant difference was observed in the HUVECs survival rates between the ultrasound group with and without the addition of SonoVue (94.1% ± 2.3% vs 91.1% ± 4.1% ). CONCLUSION: The cell membrane permeability of HUVECs and the transfection efficiency of pEGFP into HUVECs exposed to ultrasound are significantly increased after addition of an ultrasound contrast agent without obvious damage to the survival of HUVECs. This non- invasive gene transfer method may be a useful tool for clinical gene therapy of hepatic tumors. 展开更多
关键词 MICROBUBBLE ULTRASOUND Gene transfer Human umbilical vein endothelial cell Enhanced green fluorescent protein
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Synthesis and biochemical properties of fluorescent/magnetic bifunctional starch particles 被引量:1
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作者 王凤华 刘俊 +6 位作者 唐冬英 薛昌刚 肖苏尧 郑元青 童春义 王玲玲 刘选明 《Journal of Central South University》 SCIE EI CAS 2010年第2期211-217,共7页
Magnetic starch particles (MSPs) were synthesized in water-in-oil mieroemulsion at room temperature. MSPs were characterized by transmission electron microscopy (TEM), Fourier transform infrared spectrometry (FTI... Magnetic starch particles (MSPs) were synthesized in water-in-oil mieroemulsion at room temperature. MSPs were characterized by transmission electron microscopy (TEM), Fourier transform infrared spectrometry (FTIR), zeta potential system, thermogravimetric analysis (TGA) and vibrating sample magnetometry (VSM). The average diameter of the MSPs was 220 nm, dispersed with well-proportioned size and magnetic resonance, the saturation magnetization was 3.64 A.mR/kg. MSP was coated with poly-L-lysine (PLL), and then the surface of PLL-MSP was combined with fluorescein isothiocynate (FITC). Results show that fluorescent/magnetic starch particles (FMSPs) are of stable photo-bleaching capability compared with free FITC, with low bio-toxicity and certain function of magnetic separation. It is expected that FMSPs are bifimctional nano-materials including fluorescence labelling and magnetic separation. 展开更多
关键词 starch-particles NANO-MATERIALS SYNTHESIS fluorescent properties magnetic properties biochemical properties
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Fabricating Nanogaps between Nanoelectrodes using Dielectrophoresis Technique for Molecular Fluorescence Enhancement
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作者 蔡洪冰 林珂 +4 位作者 朱三娥 廖源 张杨 王晓平 董振超 《Chinese Journal of Chemical Physics》 SCIE CAS CSCD 2016年第3期279-283,I0001,共6页
Here we demonstrate the fabrication of nanometer-sized gaps by assembling single coreshell nanoparticles between metallic nanoelectrodes. Protein coated SiO2@Au coreshell nanopar- tides arc synthesized and positioned ... Here we demonstrate the fabrication of nanometer-sized gaps by assembling single coreshell nanoparticles between metallic nanoelectrodes. Protein coated SiO2@Au coreshell nanopar- tides arc synthesized and positioned between fluorescent molecules-covered electrodes in a controllable way using dielectrophoretic trapping, forming nanogaps sandwiched between nanoparticle and manoelectrodes. Preliminary photoluminescence measurements show that enhanced molecular fluorescence could be detected from the fluorescent molecules inside the nanogaps. These results pave the way for realizing electrically driven molecular fluorescence based on nanogap electrodes. 展开更多
关键词 NANOGAP NANOELECTRODE Dielectrophoresis Coreshell nanoparticle Fluores-cence enhancement
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Development of a real-time PCR assay(SYBR Green I) for rapid identification and quantification of scyphomedusae Aurelia sp.1 planulae 被引量:2
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作者 王建艳 甄毓 +2 位作者 米铁柱 于志刚 王国善 《Chinese Journal of Oceanology and Limnology》 SCIE CAS CSCD 2015年第4期974-987,共14页
The complicated life cycle ofAurelia spp., comprising benthic asexually-reproducing polyps and sexually-reproducing medusae, makes it hard for researchers to identify and track them, especially for early stage individ... The complicated life cycle ofAurelia spp., comprising benthic asexually-reproducing polyps and sexually-reproducing medusae, makes it hard for researchers to identify and track them, especially for early stage individuals, such as planulae. To solve this problem, we developed a real-time PCR assay (SYBR Green I) to identify planulae in both cultured and natural seawater samples. Species-specific primers targeting Aurelia sp.1 mitochondrial 16S rDNA (mr 16S rDNA) regions were designed. Using a calibration curve constructed with plasmids containing the Aurelia sp. 1 mt 16S rDNA fragment and a standard curve for planulae, the absolute number of mt 16S rDNA copies per planula was determined and from that the total number ofplanulae per sample was estimated. For the field samples, a 100-fold dilution of the sample DNA combined with a final concentration of 0.2 μg/μL BSA in the PCR reaction mixture was used to remove real- time PCR inhibitors. Samples collected in Jiaozhou Bay from July to September 2012 were subsequently analyzed using this assay. Peak Aurelia sp.1 planula abundance occurred in July 2012 at stations near Hongdao Island and Qingdao offshore; abundances were very low in August and September. The real-time PCR assay (SYBR Green I) developed here negates the need for traditional microscopic identification, which is laborious and time-consuming, and can detect and quantify jellyfish planulae in field plankton samples rapidly and specifically. 展开更多
关键词 Aurelia sp. 1 16S rDNA planulae real-time PCR jellyfish blooms
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