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The construction of drug-resistant cancer cell lines by CRISPR/Cas9 system for drug screening

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Science Bulletin 2018年第21期63卷 1411-1419页

作者： Lingmin Zhang Ying Li Qinghua Chen Yong Xia wenfu zheng Xingyu Jiang CAS Center for Excellence in Nanoscience Beijing Engineering Research Center for BioNanotechnologyCAS Key Laboratory for Biological Effects of Nanomaterials and NanosafetyNational Center for NanoScience and TechnologyBeijing 100190China University of Chinese Academy of Sciences Beijing 100049China Guangdong Provincial Key Laboratory of Molecular Target &Clinical Pharmacology School of Pharmaceutical Sciences and the Third &Fifth Affiliated HospitalGuangzhou Medical UniversityGuangzhou 511436China

Cancer therapy is often hampered by the rapid emergence of drug resistance. Drug-resistant cellular models are essential for understanding the drug resistance and developing new therapeutics. The low efficiency and long time required in creating these models are major obstacles hindering drug resistance research and drug screening. Herein, we report an approach that can accelerate(shortening the time from years to 3 weeks) the establishment of cancer cell line-based, inheritable drug resistance models by specific knockout of MED12 gene using CRISPR/Cas9 system. The resultant MED12^(KO) A375(melanoma)cell line was resistant to inhibitors of B-Raf proto-oncogene, serine/threonine kinase(BRAF), whereas the resultant MED12^(KO) PC9(non-small cell lung cancer) cell line was resistant to inhibitors of epidermal growth factor receptor(EGFR). Evaluation of anti-cancer drugs and their combinations shows that certain combinations of BRAF inhibitors and TGF-β receptor(TGF-βR) inhibitors are active in suppressing the growth of MED12^(KO) A375 cells, and a few combinations of EGFR inhibitors and TGF-βR inhibitors were active in suppressing the growth of MED12^(KO) PC9 cells. The drug-resistant models will be useful in screening novel drugs and drug combinations for multi-drug-resistant cancer therapy.

关键词： CRISPR/Cas9 Drug resistance Cancer Cell line Gene editing

Microscale methods to assemble mammalian cells into tissue-like structures

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Science China(Life Sciences) 2012年第10期55卷 862-871页

作者： GONG PeiYuan zheng wenfu XIAO Dan JIANG XingYu College of Chemistry Sichuan UniversityChengdu 610064China Key Lab for Biological Effects of Nanomaterials and Nanosafety National Center for Nanoscience and TechnologyBeijing 100190China

Different cell types make up tissues and organs hierarchically and communicate within a complex, three-dimensional (3D) en- vironment. The in vitro recapitulation of tissue-like structures is meaningful, not only for fundamental cell biology research, but also for tissue engineering (TE). Currently, TE research adopts either the top-down or bottom-up approach. The top-down approach involves defining the macroscopic tissue features using biomaterial scaffolds and seeding cells into these scaffolds. Conversely, the bottom-up approach aims at crafting small tissue building blocks with precision-engineered structural and functional microscale features, using physical and/or chemical approaches. The bottom-up strategy takes advantage of the repeating structural and functional units that facilitate cell-cell interactions and cultures multiple cells together as a functional unit of tissue. In this review, we focus on currently available microscale methods that can control mammalian cells to assemble into 3D tissue-like structures.

关键词： microscale cell assembly tissue-like structures tissue engineering

Change of substrate geometry and laminin density stimulates axon branching

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医用生物力学 2013年第S1期28卷 116-117页

作者： Wenwen Liu wenfu zheng Xingyu Jiang CAS Key Lab for Biological Effects of Nanomaterials and Nanosafety National Center for NanoScience and Technology

Axon branching enables neurons to contact with multiple targets and respond to their *** to its importance in neuronal network formation,axon branching has been studied extensively during the past *** is reported that ECM(Extra Cellular Matrix)components such as laminin,collagen,and tenascin regulate the morphology and motility of neuronal growth cones in culture,but the effects of their distribution and the change of density on axon branching are not well *** fabricated chemically homogeneous substrate by microcontact printing(μCP)and inhomogeneous substrate with different laminin density

关键词： laminin branching neuronal inhomogeneous chemically understood collagen motility regulate cones

A zwitterionic polymer-inspired material mediated efficient CRISPR-Cas9 gene editing

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Asian Journal of Pharmaceutical Sciences 2022年第5期17卷 666-678页

作者： Lingmin Zhang Langyu Yang Jionghua Huang Sheng Chen Chuangjia Huang Yinshan Lin Ao Shen ZhouYikang zheng wenfu zheng Shunqing Tang Key Laboratory of Molecular Target&Clinical Pharmacology and the State&NMPA Key Laboratory of Respiratory Disease School of Pharmaceutical Sciences&The Third and Fifth Affiliated HospitalGuangzhou Medical UniversityGuangzhou 511436China Department of Biomedical Engineering Jinan UniversityGuangzhou 510632China CAS Key Lab for Biological Effects of Nanomaterials and Nanosafety National Center for NanoScience and TechnologyBeijing 100190China

The typeⅡ prokaryotic CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR/Cas9) adaptive immune system is a cutting-edge genome-editing ***,its applications are still limited by its inefficient ***,we present a novel gene vector,the zwitterionic polymer-inspired material with branched structure (ZEBRA) for efficient CRISPR/Cas9 ***-like kinase 1 (PLK1) acts as a master regulator of mitosis and overexpresses in multiple tumor *** Cas9 and single guide sgRNA (sgRNA)-encoded plasmid was transduced to knockout Plk1 gene,which was expected to inhibit the expression of *** studies demonstrated that ZEBRA enabled to transduce the CRISPR/Cas9 system with large size into the cells *** transduction with ZEBRA was cell line dependent,which showed~10-fold higher in CD44-positive cancer cell lines compared with CD44-negative ***,ZEBRA induced highlevel expression of Cas9 proteins by the delivery of CRISPR/Cas9 and efficient gene editing of Plk1 gene,and inhibited the tumor cell growth *** zwitterionic polymerinspired material is an effective and targeted gene delivery vector and further studies are required to explore its potential in gene delivery applications.

关键词： CRISPR/Cas9 Gene editing Zwitterionic polymers CD44 PLK1

Nanoliposome-encapsulated caged-GABA for modulating neural electrophysiological activity with simultaneous detection by microelectrode arrays

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Nano Research 2020年第6期13卷 1756-1763页

作者： Jingyu Xie Yilin Song Yuchuan Dai Ziyue Li Fei Gao Xuanyu Li Guihua Xiao Yu Zhang Hao Wang Zeying Lu Xingyu Jiang wenfu zheng Xinxia Cai State Key Laboratory of Transducer Technology Institute of ElectronicsChinese Academy of SciencesBeijing100190China University of Chinese Academy of Sciences Beijing100049China Beijing Engineering Research Center for BioNanotechnology CAS Key Laboratory for Biological Effects of Nanomaterials and NanosafetyCAS Center for Excellence in NanoscienceNational Center for NanoScience and TechnologyBeijing100190China Department of Biomedical Engineering Southern University of Science and TechnologyShenzhen518055China

Effective and precise neural modulation with real-time detection in the brain is of great importance and represents a significant ***-encapsulated light-sensitive compounds have excellent characteristics such as high temporal and spatial resolution,delayed drug clearance,and restricted drug biodistribution for neural *** this study,we developed a nanoliposome-based delivery system for ruthenium-based caged GABA compounds(Nanolipo-Ru)to modulate neural activity and allow for real-time monitoring using the microelectrode arrays(MEAs).The Nanolipo-Ru nanoparticles had an average size of 134.10±4.30 nm and exhibited excellent stability for seven *** the in vivo experiment in the rat,release of GABA by Nanolipo-Ru under blue light illumination resulted in an average firing rate reduction in interneurons and pyramidal neurons in the same brain region of 79.4%and 81.6%,***,the average power of local field potentials in the 0–15 Hz range degraded from 4.34 to 0.85 *** addition,the Nanolipo-Ru nanoparticles have the potential to provide more flexible timing of modulation than unencapsulated RuBi-GABA in the *** results indicated that Nanolipo-Ru could be an effective platform for regulating neuronal ***,nanoliposomes with appropriate modifications would render promising utilities for targeting of specific types of neurons in the future.

关键词： neural modulation nanoliposome microelectrode array light-sensitive drug

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直感决策的参考点方法研究

系统工程理论与实践 1999年第1期19卷 51-55,111页

作者：郑文富李怀祖西安交通大学管理学院

高层管理者的直感决策常常建立在一定的参考点基础之上，直感决策的研究涉及决策者选择合适参考点的思维机理．本文定义并区分了四类直感决策参考点：低值参考点、高值参考点、匹配型参考点及背景型参考点，讨论了决策者在分别运用这四... 详细信息

高层管理者的直感决策常常建立在一定的参考点基础之上，直感决策的研究涉及决策者选择合适参考点的思维机理．本文定义并区分了四类直感决策参考点：低值参考点、高值参考点、匹配型参考点及背景型参考点，讨论了决策者在分别运用这四类决策参考点时可能引起的偏差．

关键词：直感决策相似效应参考点效应高层管理者

新时代优化开发区绿色经济绩效评估及时空演变--基于DDF-Global Malmquist-Luenberger指数方法分析

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广西财经学院学报 2020年第2期33卷 69-80页

作者：岳立郑文富兰州大学经济学院甘肃兰州730000

新时代背景下,传统的绩效考核方式对促进经济高质量发展的作用有限,需转变考核方式。作为主体功能区重要组成部分的优化开发区是体现中国经济实力的重要区域,其资源环境等问题也更为突出,开展对其的绿色经济绩效评估对推动中国经济高质... 详细信息

新时代背景下,传统的绩效考核方式对促进经济高质量发展的作用有限,需转变考核方式。作为主体功能区重要组成部分的优化开发区是体现中国经济实力的重要区域,其资源环境等问题也更为突出,开展对其的绿色经济绩效评估对推动中国经济高质量发展具有重要作用。基于优化开发区的视角,以其中的54个城市作为研究对象,运用DDF-GML指数模型评价其2007-2015年考虑非期望产出的绿色经济绩效和未考虑非期望产出的传统经济绩效,分析二者的时空演变特征,探讨优化开发区绿色经济绩效的提升途径。研究发现:开发区传统经济绩效评价低估了实际绩效情况,从横向区域角度分析,绩效呈由北向南递减的趋势,绩效及其动力源泉具有区域异质性,绿色经济绩效和传统经济绩效差异较为明显的城市来自珠江三角洲和环渤海地区;从纵向时间维度分析,技术效率已成为开发区绩效更快提升的主要约束,此外,长江三角洲绩效增长率震荡幅度最大。为此,优化开发区各城市应优化绿色绩效评价指标,加强生态环境的保护作用;调整优化产业结构,注重资源的集约化利用,改善技术效率,从而提升优化开发区的绿色经济绩效。

关键词：优化开发区绿色经济绩效绩效评价 GML指数

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爆炸挤淤在填海造路软基处理工程中应用

土工基础 2023年第5期37卷 707-711页

作者：张文龙郑文富湛江市矿大爆破有限公司广东湛江524000 广东中人爆破工程有限公司广州510000

在软基处理中,爆炸挤淤具有施工便捷、节约成本、工期短、软基处理效果显著等特点。以宁德市某填海造路工程为研究对象,采用干滩(即滩涂爆破)爆炸挤淤法代替原砂桩及管桩处理软基法,根据该工程施工特点、淤泥物理力学性质和施工难点,设... 详细信息

在软基处理中,爆炸挤淤具有施工便捷、节约成本、工期短、软基处理效果显著等特点。以宁德市某填海造路工程为研究对象,采用干滩(即滩涂爆破)爆炸挤淤法代替原砂桩及管桩处理软基法,根据该工程施工特点、淤泥物理力学性质和施工难点,设计了该填海工程软基处理的爆炸挤淤方案和技术参数,阐述了可能造成的爆破危害效应及相应的安全控制措施。监测表明,采用该方法爆破后堤身稳定性好,爆破挤淤满足工后月沉降不大于6 mm的沉降标准,爆破挤淤法处理软基的方式是科学可行的,为其他类似工程提供借鉴。

关键词：爆破挤淤填海工程软基处理安全措施

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框架核心筒结构高层建筑的对向倒塌爆破拆除

爆破器材 2022年第6期51卷 60-64页

作者：杨志红郑文富何慧明陈亚伟广东中人爆破工程有限公司广东广州510640

介绍了城市复杂环境中采用双切口对向倒塌爆破拆除两栋高层框架核心筒结构大楼的工程实例。根据框架核心筒结构特点,结合场地周边环境,选择对向倒塌的爆破方案。先爆建筑采用双切口方式,后爆建筑采用单切口方式,通过四通与簇联相结合的... 详细信息

介绍了城市复杂环境中采用双切口对向倒塌爆破拆除两栋高层框架核心筒结构大楼的工程实例。根据框架核心筒结构特点,结合场地周边环境,选择对向倒塌的爆破方案。先爆建筑采用双切口方式,后爆建筑采用单切口方式,通过四通与簇联相结合的复式起爆网路,楼间间隔3 s延时起爆,实现大楼逐栋倒塌解体;采用主动防护与被动防护相结合的安全防护技术,有效地控制了爆破振动、塌落振动和爆破飞散物,取得了良好的爆破效果。

关键词：框架核心筒结构对向倒塌双切口爆破拆除