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Real-time cell analysis and heat shock protein gene expression in the TcA Tribofium castaneum cell line in response to environmental stress conditions 被引量:3
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作者 Andres Garcia-Reina Maria Juliana Rodriguez-Garcia +1 位作者 Guillermo Ramis Jose Galian 《Insect Science》 SCIE CAS CSCD 2017年第3期358-370,共13页
The rust red flour beetle, Tribolium castaneum (Herbst, 1797) (Coleoptera: Tenebrionidae), is a pest of stored grain and one of the most studied insect model species. Some of the previous studies involved heat re... The rust red flour beetle, Tribolium castaneum (Herbst, 1797) (Coleoptera: Tenebrionidae), is a pest of stored grain and one of the most studied insect model species. Some of the previous studies involved heat response studies in terms of survival and heat shock protein expression, which are regulated to protect other proteins against environ- mental stress conditions. In the present study, we characterize the impedance profile with the xCELLigence Real-Time Cell Analyzer and study the effect of increased temperature in cell growth and viability in the cell line BCIRL-TcA-CLG 1 (TcA) of T castaneum. This novel system measures cells behavior in real time and is applied for the first time to insect cells. Additionally, cells are exposed to heat shock, increased salinity, acidic pH and UV-A light with the aim of measuring the expression levels of lisp27, Hsp68a, and Hsp83 genes. Results show a high thermotolerance of TeA in terms of cell growth and viability. This result is likely related to gene expression results in which a significant up-regulation of all studied Hsp genes is observed after 1 h of exposure to 40 ~C and UV light. All 3 genes show similar expression patterns, but Hsp27 seems to be the most affected. The results of this study validate the RTCA method and reveal the utility of insect cell lines, real-time analysis and gene expression studies to better understand the physiological response of insect cells, with potential applications in different fields of biology such as conservation biology and pest management. 展开更多
关键词 environmental stress gene expression heat shock proteins insect cells real-time cell analysis (rtca red flour beetle
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A novel method for evaluating the dynamic biocompatibility of degradable biomaterials based on real-time cell analysis 被引量:5
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作者 Xiaoxiao Gai Chenghu Liu +4 位作者 Guowei Wang Yang Qin Chunguang Fan Jia Liu Yanping Shi 《Regenerative Biomaterials》 SCIE 2020年第3期321-329,共9页
Degradable biomaterials have emerged as a promising type of medical materials because of their unique advantages of biocompatibility,biodegradability and biosafety.Owing to their bioabsorbable and biocompatible proper... Degradable biomaterials have emerged as a promising type of medical materials because of their unique advantages of biocompatibility,biodegradability and biosafety.Owing to their bioabsorbable and biocompatible properties,magnesium-based biomaterials are considered as ideal degradable medical implants.However,the rapid corrosion of magnesium-based materials not only limits their clinical application but also necessitates a more specific biological evaluation system and biosafety standard.In this study,extracts of pure Mg and its calcium alloy were prepared using different media based on ISO 10993:12;the Mg^2+ concentration and osmolality of each extract were measured.The biocompatibility was investigated using the MTT assay and xCELLigence real-time cell analysis(RTCA).Cytotoxicity tests were conducted with L929,MG-63 and human umbilical vein endothelial cell lines.The results of the RTCA highly matched with those of the MTT assay and revealed the different dynamic modes of the cytotoxic process,which are related to the differences in the tested cell lines,Mg-based materials and dilution rates of extracts.This study provides an insight on the biocompatibility of biodegradable materials from the perspective of cytotoxic dynamics and suggests the applicability of RTCA for the cytotoxic evaluation of degradable biomaterials. 展开更多
关键词 degradable biomaterials BIOCOMPATIBILITY real-time cell analysis magnesium-based biomaterials
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Synergistic cytotoxicity of binary combinations of inorganic and organic disinfection byproducts assessed by real-time cell analysis
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作者 Mengge Fan Longfei Shu +4 位作者 Xinran Zhang Miao Yu Yongting Du Junlang Qiu Xin Yang 《Journal of Environmental Sciences》 SCIE EI CAS CSCD 2022年第7期222-231,共10页
Chlorine, chlorine dioxide, and ozone are widely used as disinfectants in drinking water treatments. However, the combined use of different disinfectants can result in the formation of various organic and inorganic di... Chlorine, chlorine dioxide, and ozone are widely used as disinfectants in drinking water treatments. However, the combined use of different disinfectants can result in the formation of various organic and inorganic disinfection byproducts(DBPs). The toxic interactions, including synergism, addition, and antagonism, among the complex DBPs are still unclear. In this study, we established and verified a real-time cell analysis(RTCA) method for cytotoxicity measurement on Chinese hamster ovary(CHO) cell. Using this convenient and accurate method, we assessed the cytotoxicity of a series of binary combinations consisting of one of the 3 inorganic DBPs(chlorite, chlorate, and bromate) and one of the 32 regulated and emerging organic DBPs. The combination index(CI) of each combination was calculated and evaluated by isobolographic analysis to reflect the toxic interactions. The results confirmed the synergistic effect on cytotoxicity in the binary combinations consisting of chlorite and one of the 5 organic DBPs(2 iodinated DBPs(I-DBPs) and 3 brominated DBPs(Br-DBPs)), chlorate and one of the 4 organic DBPs(3 aromatic DBPs and dibromoacetonitrile), and bromate and one of the 3 organic DBPs(2 I-DBPs and dibromoacetic acid). The possible synergism mechanism of organic DBPs on the inorganic ones may be attributed to the influence of organic DBPs on cell membrane and cell antioxidant system. This study revealed the toxic interactions among organic and inorganic DBPs, and emphasized the latent adverse outcomes in the combined use of different disinfectants. 展开更多
关键词 Disinfection byproducts(DBPs) real-time cell analysis(rtca) Synergistic cytotoxicity
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纳米氧化铜作用于CHO细胞的细胞毒性实时动态研究
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作者 段链 顾雯 张宏伟 《环境卫生学杂志》 2015年第5期403-407,413,共6页
目的对纳米氧化铜作用于CHO细胞在连续时间点所产生的细胞毒性效应进行研究,得到准确数据,为进一步毒性机制研究提供参考依据。方法应用RTCA(real-time cell analysis)对不同密度的CHO细胞进行长时间连续测定,绘制不同密度CHO细胞的... 目的对纳米氧化铜作用于CHO细胞在连续时间点所产生的细胞毒性效应进行研究,得到准确数据,为进一步毒性机制研究提供参考依据。方法应用RTCA(real-time cell analysis)对不同密度的CHO细胞进行长时间连续测定,绘制不同密度CHO细胞的完整生长曲线。对不同剂量的纳米氧化铜作用于CHO细胞所产生的毒性效应进行实时监测,得到不同作用时间的准确IC50值,绘制连续时间点IC50变化曲线,以判定纳米氧化铜作用于CHO细胞所产生的毒性效应。结果 4种不同密度细胞接种后对数生长期时间有所不同,其中5×10^3个/孔密度最适合进行毒性效应试验。不同剂量纳米氧化铜染毒后,毒性作用起始时间约4~5 h,作用12、24、36和48 h的IC50值分别为0.249、0.207、0.236和0.412 mg/m L。结论纳米氧化铜作用于CHO细胞,毒性作用起始时间约为染毒后3 h开始,作用6 h后,IC50值明显下降,且随着时间延长呈现下降趋势。作用时间达到18 h后,IC50值持续在低于0.2 mg/m L以下,且数值接近。 展开更多
关键词 纳米氧化铜 CHO细胞 rtca实时监测
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shRNA沉默HuR抑制肝细胞癌细胞系SMMC-7721的增殖、迁移及侵袭能力 被引量:4
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作者 湛钊 周莉莉 +4 位作者 高义沙 余畅 陈相屹 胡桐 孙达权 《中国生物化学与分子生物学报》 CAS CSCD 北大核心 2020年第8期952-960,共9页
人抗原R (human antigen R,HuR)基因在肺癌、乳腺癌等多种肿瘤组织中高表达。推测HuR基因也参与肝癌的发展过程。为探索HuR对肝细胞癌细胞系SMMC-7721的增殖、迁移和侵袭的作用,本研究通过蛋白质印迹实验,检测HuR在肝细胞癌细胞系和正... 人抗原R (human antigen R,HuR)基因在肺癌、乳腺癌等多种肿瘤组织中高表达。推测HuR基因也参与肝癌的发展过程。为探索HuR对肝细胞癌细胞系SMMC-7721的增殖、迁移和侵袭的作用,本研究通过蛋白质印迹实验,检测HuR在肝细胞癌细胞系和正常肝细胞中蛋白质的表达水平。结果显示,肝细胞癌细胞系SMMC-7721 HuR的表达量显著高于正常肝细胞HL-7702。合成特异性靶向HuR基因的shRNA,转染肝细胞癌细胞系SMMC-7721,检测结果发现,HuR基因表达量下调90%。沉默HuR,实时细胞分析技术(real time cell analysis,RTCA)结果显示,细胞增殖能力降低55%,侵袭能力下降75%;细胞划痕实验结果显示,迁移能力下降80%;克隆形成实验中细胞克隆数减少85%;此外,在HuR敲低稳转肝细胞癌细胞系SMMC-7721细胞中过表达Bcl-2,其细胞学现象部分获得逆转。激光共聚焦扫描系统检测结果显示,Bcl-2主要定位于肝细胞癌细胞系SMMC-7721的核膜及胞质。蛋白质印迹法检测结果显示,沉默HuR,Bcl-2下调92%,Survivin下调55%,Twinst1下调69%,N-钙黏着蛋白(N-cadherin)下调48%,E-钙黏着蛋白(E-cadherin)上调1.5倍。以上结果表明,HuR可能通过调控定位于核膜及胞质上的Bcl-2来参与肝细胞癌细胞系SMMC-7721的增殖、迁移、侵袭及克隆形成过程,HuR基因有望成为临床上治疗肝癌的一个新的潜在靶点。 展开更多
关键词 人抗原R SMMC-7721 实时细胞分析技术 增殖 激光共聚焦系统
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Review of Some Advances and Applications in Real-time High-speed Vision: Our Views and Experiences 被引量:2
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作者 Qing-Yi Gu Idaku Ishii 《International Journal of Automation and computing》 EI CSCD 2016年第4期305-318,共14页
The frame rate of conventional vision systems is restricted to the video signal formats (e.g., NTSC 30 fps and PAL 25 fps) that are designed on the basis of the characteristics of the human eye, which implies that t... The frame rate of conventional vision systems is restricted to the video signal formats (e.g., NTSC 30 fps and PAL 25 fps) that are designed on the basis of the characteristics of the human eye, which implies that the processing speed of these systems is limited to the recognition speed of the human eye. However, there is a strong demand for real-time high-speed vision sensors in many application fields, such as factory automation, biomedicine, and robotics, where high-speed operations are carried out. These high-speed operations can be tracked and inspected by using high-speed vision systems with intelligent sensors that work at hundreds of Hertz or more, especially when the operation is difficult to observe with the human eye. This paper reviews advances in developing real-time high Speed vision systems and their applications in various fields, such as intelligent logging systems, vibration dynamics sensing, vision-based mechanical control, three-dimensional measurement/automated visual inspection, vision-based human interface, and biomedical applications. 展开更多
关键词 real-time high-speed vision target tracking abnormal behavior detection behavior mining vibration analysis 3D shapemeasurement cell sorting.
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基于实时细胞分析技术评价注射用益气复脉(冻干)类过敏反应 被引量:4
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作者 范姗姗 陈瑞 +6 位作者 尹清晟 韩娟 张艳军 周大铮 鞠爱春 李德坤 庄朋伟 《药物评价研究》 CAS 2018年第3期411-416,共6页
目的建立中药注射剂体外类过敏反应评价方法,快速评价不同批次注射用益气复脉(冻干)类过敏反应表现。方法体外培养嗜碱性白血病细胞株RBL-2H3细胞,选择Compound 48/80为阳性药,采用实时细胞分析(real-time cell analysis,RTCA)系统检测... 目的建立中药注射剂体外类过敏反应评价方法,快速评价不同批次注射用益气复脉(冻干)类过敏反应表现。方法体外培养嗜碱性白血病细胞株RBL-2H3细胞,选择Compound 48/80为阳性药,采用实时细胞分析(real-time cell analysis,RTCA)系统检测药物干预后引起的细胞指数(CI)值变化,并利用甲苯胺蓝、鬼笔环肽染色观察细胞形态、骨架变化,以及检测组胺和β-已糖苷酶释放量验证RBL-2H3细胞的脱颗粒情况。选择20个批次的注射用益气复脉(冻干,100μg/m L)作用于RBL-2H3细胞,进行基于RTCA技术的类过敏反应评价。结果阳性药Compound 48/80(20μg/m L)能够使RBL-2H3细胞的CI值在加药后30 min内呈先快速上升后下降趋势;形态学研究发现,Compound 48/80使细胞形态和细胞骨架均发生明显改变,发生明显的脱颗粒现象;组胺和β-己糖苷酶释放实验进一步证实Compound 48/80导致炎症介质的释放,引起了明显的脱颗粒现象;提示RTCA系统可以用于快速敏感的评价RBL-2H3细胞脱颗粒。不同批次的注射用益气复脉(冻干)对RBL-2H3细胞CI值无明显影响,提示所选批次为合格批次,无类过敏反应现象的发生。结论建立了一套基于RTCA系统的类过敏反应体外快速评价技术,可用于注射用益气复脉(冻干)等中药注射剂类过敏反应的体外快速评价。 展开更多
关键词 实时细胞分析(rtca) Compound 48/80 注射用益气复脉(冻干) 类过敏反应 RBL-2H3细胞 脱颗粒
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基于实时细胞分析技术验证人诱导多能干细胞分化的心肌细胞毒性评价模型
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作者 陈高建 潘东升 +6 位作者 陈思蓉 张颖丽 李芊芊 石茜茜 王雪 黄芝瑛 王三龙 《药物分析杂志》 CAS CSCD 北大核心 2020年第1期90-103,共14页
目的:利用人诱导多能干细胞分化的心肌细胞(hiPSC-CMs)联合实时细胞分析(RTCA)技术,建立hiPSC-CMs体外心脏毒性评价模型,并选用阳性化合物进行验证。方法:研究选用商业化来源的hiPSC-CMs,选用不同作用机制的心脏毒性药物:蒽环类抗癌药... 目的:利用人诱导多能干细胞分化的心肌细胞(hiPSC-CMs)联合实时细胞分析(RTCA)技术,建立hiPSC-CMs体外心脏毒性评价模型,并选用阳性化合物进行验证。方法:研究选用商业化来源的hiPSC-CMs,选用不同作用机制的心脏毒性药物:蒽环类抗癌药阿霉素(0.3、1、3μmol·L^-1)、K+通道阻滞剂多非利特(0.03、0.1、0.3μmol·L^-1)和E-4031(0.1、0.3、1μmol·L^-1)、Na+通道阻滞剂美西律(3、10、30μmol·L^-1)、多离子通道阻滞剂维拉帕米(0.1、0.3、1μmol·L^-1)和苄普地尔(0.3、1、3μmol·L^-1)、抗组胺药物特非那定(0.3、1、3μmol·L^-1)和西沙必利(3、10、30μmol·L^-1),分别给予hiPSC-CMs作用48 h,采用RTCA技术连续监测心肌细胞搏动的频率、振幅、细胞指数、搏动图谱等指标在给药前后的变化。结果:阿霉素对心肌细胞的抑制作用呈现时效和量效依赖性特点;多非利特显著降低心肌细胞搏动幅度及频率;E-4031为1μmol·L^-1时显著降低心肌细胞搏动频率,缩短振幅,而0.1、0.3μmol·L^-1时则对心肌细胞无影响;30μmol·L^-1美西律作用2 h内心肌细胞搏动频率相对空白对照组降低了66.4%,振幅缩短了60.7%,并导致心肌细胞搏动周期延长;苄普地尔浓度低于1μmol·L^-1时对心肌细胞无影响,3μmol·L^-1苄普地尔作用2 h导致心肌细胞搏动间歇性停搏,并伴随细胞活力轻微下降;维拉帕米作用2 h内各剂量组心肌细胞搏动频率分别增强了12.2%、13.1%、52.3%,而振幅分别降低了20.0%、61.7%、67.6%。0.3μmol·L^-1以上浓度的维拉帕米导致心肌细胞搏动骤停(3/9);特非那定浓度≥1μmol·L^-1时短时间内即导致心肌细胞间歇性搏动骤停;西沙必利短期作用2 h内显著降低心肌细胞搏动频率和幅度,10μmol·L^-1以上浓度的西沙必利持续抑制心肌细胞搏动频率。结论:hiPSC-CMs结合RTCA技术可用于预测药物的心脏毒性风险。 展开更多
关键词 诱导多能干细胞分化的心肌细胞 心律失常 实时细胞分析(rtca) 综合性离体致心律失常风险评估(CiPA) 频率 振幅
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苯并[a]芘的细胞毒性及诱导IL-1β和IL-8表达作用研究
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作者 陈丝秦 银星月 +3 位作者 何庆华 严茂胜 王知源 杨彬珧 《中国工业医学杂志》 CAS 2022年第2期105-109,183,共6页
目的探讨不同浓度苯并[a]芘(BaP)对人肺癌(淋巴结转移)NCI-H292的细胞毒性及其对NCI-H292细胞炎性因子分泌的影响。方法取对数期生长的NCI-H292细胞,分别以不同剂量BaP(0、4、8、16和32μmol/L)染毒24、48和72 h后,采用实时无标记细胞... 目的探讨不同浓度苯并[a]芘(BaP)对人肺癌(淋巴结转移)NCI-H292的细胞毒性及其对NCI-H292细胞炎性因子分泌的影响。方法取对数期生长的NCI-H292细胞,分别以不同剂量BaP(0、4、8、16和32μmol/L)染毒24、48和72 h后,采用实时无标记细胞分析系统(RTCA)法检测细胞存活率,ELISA法检测细胞内白介素-1β(IL-1β)和白介素-8(IL-8)水平。本实验按照5×3析因设计。结果与对照组相比,各剂量BaP染毒组在24、48和72 h时间点细胞存活率均显著降低(P<0.001),细胞存活率与染毒剂量之间呈高度负相关关系(r;=-0.986、-0.974和-0.993,P<0.01);与同剂量组24 h时比较,各实验组在染毒48 h和72 h细胞存活率均显著升高(P<0.001),细胞存活率与染毒时间之间呈高度正相关关系(r;=0.958、0.996、0.994、0.999和1.000,P<0.01)。NCI-H292细胞内IL-1β水平在染毒剂量和染毒时间上有交互效应(P<0.001),在48 h和72 h时BaP高剂量(8~32μmol/L)染毒组IL-1β水平均呈随着染毒剂量升高而降低(P<0.01),72 h时低剂量BaP染毒组IL-1β水平则呈随着染毒剂量升高而升高的剂量-效应关系(P<0.01);同时,各实验组细胞IL-1β水平在24、48和72 h均呈随着染毒时间增加而上升的时间-效应关系(P<0.01)。NCI-H292细胞内IL-8水平在染毒剂量和染毒时间上有交互效应(P<0.01):在染毒72 h,BaP染毒剂量为0~16μmol/L时IL-8水平呈随着染毒剂量升高而升高的剂量-效应关系(P<0.01);4、8μmol/L染毒组细胞IL-8水平在24、48和72 h呈随着染毒时间增加而上升的时间-效应关系(P<0.01)。结论BaP染毒可引起NCI-H292细胞的存活率下降,且随着染毒剂量增加毒性作用增强;BaP可刺激NCI-H292细胞分泌IL-1β和IL-8,其可能在BaP致肺肿瘤炎症过程中发挥重要作用。 展开更多
关键词 苯并[a]芘(BaP) NCI-H292细胞 细胞毒性 实时无标记细胞分析法(rtca) 白介素-1β(IL-1β) 白介素-8(IL-8)
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