Although the recent advances in stem cell engineering have gained a great deal of attention due to their high potential in clinical research,the applicability of stem cells for preclinical screening in the drug discov...Although the recent advances in stem cell engineering have gained a great deal of attention due to their high potential in clinical research,the applicability of stem cells for preclinical screening in the drug discovery process is still challenging due to difficulties in controlling the stem cell microenvironment and the limited availability of high-throughput systems.Recently,researchers have been actively developing and evaluating three-dimensional(3D)cell culture-based platforms using microfluidic technologies,such as organ-on-a-chip and organoid-on-a-chip platforms,and they have achieved promising breakthroughs in stem cell engineering.In this review,we start with a comprehensive discussion on the importance of microfluidic 3D cell culture techniques in stem cell research and their technical strategies in the field of drug discovery.In a subsequent section,we discuss microfluidic 3D cell culture techniques for high-throughput analysis for use in stem cell research.In addition,some potential and practical applications of organ-on-a-chip or organoid-on-a-chip platforms using stem cells as drug screening and disease models are highlighted.展开更多
As an essential component of the Materials Genome Initiative aiming to shorten the period of materials research and development, combinatorial synthesis and rapid characterization technologies have been playing a more...As an essential component of the Materials Genome Initiative aiming to shorten the period of materials research and development, combinatorial synthesis and rapid characterization technologies have been playing a more and more important role in exploring new materials and comprehensively understanding materials properties. In this review, we discuss the advantages of high-throughput experimental techniques in researches on superconductors. The evolution of combinatorial thin-film technology and several high-speed screening devices are briefly introduced. We emphasize the necessity to develop new high-throughput research modes such as a combination of high-throughput techniques and conventional methods.展开更多
为探究湖北襄阳地区高温大曲中芽孢杆菌(Bacillus)菌群多样性,该研究通过下载并筛选高温大曲样品芽孢杆菌相对含量>2.0%的测序数据,采用Illumina Mi Seq高通量测序技术和传统纯培养技术相结合的方法对大曲样品中芽孢杆菌多样性进行...为探究湖北襄阳地区高温大曲中芽孢杆菌(Bacillus)菌群多样性,该研究通过下载并筛选高温大曲样品芽孢杆菌相对含量>2.0%的测序数据,采用Illumina Mi Seq高通量测序技术和传统纯培养技术相结合的方法对大曲样品中芽孢杆菌多样性进行解析。结果表明,高温大曲中芽孢杆菌相对含量>2.0%的样品共有12份,包括8份白色大曲样品和4份黄色大曲样品,所有样品共获得77 825条代表性序列,1 197个操作分类单元(OTUs)。多样性分析结果表明,较之黄色大曲,白色大曲样品中芽孢杆菌的超1指数和发现物种数显著偏高(P<0.05)。OTU分析结果表明,OTU5321和OTU5291在所有样品中均存在,其在白色大曲和黄色大曲中累积包含序列分别占总序列数的49.43%和41.32%。传统纯培养技术分析结果表明,解淀粉芽孢杆菌(Bacillus amyloliquefaciens)为高温大曲中芽孢杆菌的主要可培养菌种。由此可知,白色大曲芽孢杆菌群落结构的丰富度更高,且解淀粉芽孢杆菌为高温大曲中芽孢杆菌的主要可培养菌种,为后续高温大曲中芽孢杆菌的应用及菌种选育具有指导作用。展开更多
目的基于实时细胞分析(real time cell analysis,RTCA)技术对中药挥发性组分的细胞毒性及抗人腺病毒3型(human adenovirus 3,HAdV-3)的作用进行毒/效整合分析,构建抗病毒药物高通量筛选的新策略。方法采用RTCA技术动态监测不同接种密度...目的基于实时细胞分析(real time cell analysis,RTCA)技术对中药挥发性组分的细胞毒性及抗人腺病毒3型(human adenovirus 3,HAdV-3)的作用进行毒/效整合分析,构建抗病毒药物高通量筛选的新策略。方法采用RTCA技术动态监测不同接种密度的A549细胞48 h生长曲线及10倍梯度稀释的HAdV-3感染A549细胞生长曲线,获得A549细胞最佳接种密度及HAdV-3最佳稀释浓度用于后续实验。以利巴韦林为阳性对照,基于RTCA技术采用曲线下面积(Area Under the Curve,AUC)方法对5种中药挥发性组分的细胞毒性及抗HAdV-3的作用进行毒/效整合分析,并与传统终点法数据处理进行对比。结果终点法中艾叶、柴胡、薄荷、荆芥、牛蒡子、酸枣仁和利巴韦林对细胞的保护率分别为0.73%、12.50%、20.99%、44.50%、27.99%、51.50%和82.70%;AUC法中艾叶、柴胡、薄荷的选择性指数(Selective Index,SI)为负数,分别为-0.57、-0.21和-0.08。荆芥、牛蒡子、酸枣仁和利巴韦林的SI值分别为0.14、0.40、0.72和5.33。以利巴韦林的抗病毒活性为参照,终点法中酸枣仁、牛蒡子和荆芥的抗病毒活性为利巴韦林抗病毒活性的62.27%、33.85%和53.81%;AUC法中酸枣仁、牛蒡子和荆芥的抗病毒活性为利巴韦林抗病毒活性的13.51%、7.50%和2.63%。结论传统终点法与研究采用的AUC法计算结果有较大的差异。传统的抗病毒活性筛选研究多采用终点法检测受试药物对病毒感染细胞的保护作用,无法反映整个感染周期的变化,缺乏细胞毒的数据也会对结果的判断产生偏差。研究提示基于RTCA技术对中药挥发性组分开展抗HAdV-3的毒/效整合评价,采用AUC方法计算药物的选择性指数作为高通量筛选新策略,能快速、精准地判断受试药物的抗病毒活性,具有准确性高、重复性好的优势。展开更多
基金supported by the National Research Foundation of Korea (NRF) (NRF2017R1C1B2002377, NRF-2016R1A5A1010148, and NRF2019R1A2C1003111)funded by the Ministry of Science and ICT (MSIT)partly supported by the Technology Innovation Program (No.10067787)funded by the Ministry of Trade, Industry & Energy (MOTE, Korea)
文摘Although the recent advances in stem cell engineering have gained a great deal of attention due to their high potential in clinical research,the applicability of stem cells for preclinical screening in the drug discovery process is still challenging due to difficulties in controlling the stem cell microenvironment and the limited availability of high-throughput systems.Recently,researchers have been actively developing and evaluating three-dimensional(3D)cell culture-based platforms using microfluidic technologies,such as organ-on-a-chip and organoid-on-a-chip platforms,and they have achieved promising breakthroughs in stem cell engineering.In this review,we start with a comprehensive discussion on the importance of microfluidic 3D cell culture techniques in stem cell research and their technical strategies in the field of drug discovery.In a subsequent section,we discuss microfluidic 3D cell culture techniques for high-throughput analysis for use in stem cell research.In addition,some potential and practical applications of organ-on-a-chip or organoid-on-a-chip platforms using stem cells as drug screening and disease models are highlighted.
基金Project supported by the National Key Basic Research Program of China(Grant Nos.2015CB921000,2016YFA0300301,2017YFA0303003,and 2017YFA0302902)the National Natural Science Foundation of China(Grant Nos.11674374,11804378,and 11574372)+3 种基金the Beijing Municipal Science and Technology Project(Grant No.Z161100002116011)the Key Research Program of Frontier Sciences,Chinese Academy of Sciences(Grant Nos.QYZDB-SSW-SLH008 and QYZDY-SSW-SLH001)the Strategic Priority Research Program of the Chinese Academy of Sciences(Grant No.XDB07020100)the Opening Project of Wuhan National High Magnetic Field Center(Grant No.PHMFF2015008)
文摘As an essential component of the Materials Genome Initiative aiming to shorten the period of materials research and development, combinatorial synthesis and rapid characterization technologies have been playing a more and more important role in exploring new materials and comprehensively understanding materials properties. In this review, we discuss the advantages of high-throughput experimental techniques in researches on superconductors. The evolution of combinatorial thin-film technology and several high-speed screening devices are briefly introduced. We emphasize the necessity to develop new high-throughput research modes such as a combination of high-throughput techniques and conventional methods.
文摘为建立一种高同源区段的单核苷酸多态性(SNP)基因分型技术,通过构建本地Blast对SNP所在的200和400 bp区段进行同源性评估,并筛选出高同源区段的SNP。利用第一轮多重长PCR(polymerase chain reaction)捕获329个样本的9个高同源区段SNP所在的长片段,使用纯化后的第一轮PCR产物作为模板进行扩增子建库测序,检测样本共得2 928个SNP位点信息,测序成功率高达98.885 6%。利用Hardy-Weinberg(HWE)法则计算试验研究的9个高同源区段SNP位点的基因频率(p值均大于0.05,符合HWE法则),并与NCBI(national center for biotechnology information)中千人基因组数据库中获取的基因频率相比对,发现二者单碱基基因频率一致(误差限<0.15)。研究表明,利用多重长PCR靶向捕获技术结合二代测序技术为高同源区段的SNP分型提供一个准确、快速、大样本检测方案。
文摘目的基于实时细胞分析(real time cell analysis,RTCA)技术对中药挥发性组分的细胞毒性及抗人腺病毒3型(human adenovirus 3,HAdV-3)的作用进行毒/效整合分析,构建抗病毒药物高通量筛选的新策略。方法采用RTCA技术动态监测不同接种密度的A549细胞48 h生长曲线及10倍梯度稀释的HAdV-3感染A549细胞生长曲线,获得A549细胞最佳接种密度及HAdV-3最佳稀释浓度用于后续实验。以利巴韦林为阳性对照,基于RTCA技术采用曲线下面积(Area Under the Curve,AUC)方法对5种中药挥发性组分的细胞毒性及抗HAdV-3的作用进行毒/效整合分析,并与传统终点法数据处理进行对比。结果终点法中艾叶、柴胡、薄荷、荆芥、牛蒡子、酸枣仁和利巴韦林对细胞的保护率分别为0.73%、12.50%、20.99%、44.50%、27.99%、51.50%和82.70%;AUC法中艾叶、柴胡、薄荷的选择性指数(Selective Index,SI)为负数,分别为-0.57、-0.21和-0.08。荆芥、牛蒡子、酸枣仁和利巴韦林的SI值分别为0.14、0.40、0.72和5.33。以利巴韦林的抗病毒活性为参照,终点法中酸枣仁、牛蒡子和荆芥的抗病毒活性为利巴韦林抗病毒活性的62.27%、33.85%和53.81%;AUC法中酸枣仁、牛蒡子和荆芥的抗病毒活性为利巴韦林抗病毒活性的13.51%、7.50%和2.63%。结论传统终点法与研究采用的AUC法计算结果有较大的差异。传统的抗病毒活性筛选研究多采用终点法检测受试药物对病毒感染细胞的保护作用,无法反映整个感染周期的变化,缺乏细胞毒的数据也会对结果的判断产生偏差。研究提示基于RTCA技术对中药挥发性组分开展抗HAdV-3的毒/效整合评价,采用AUC方法计算药物的选择性指数作为高通量筛选新策略,能快速、精准地判断受试药物的抗病毒活性,具有准确性高、重复性好的优势。
