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

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 toolbox.However,its applications are still limited by its inefficient transduction.Herein,we present a novel gene vector,the zwitterionic polymer-inspired material with branched structure (ZEBRA) for efficient CRISPR/Cas9 delivery.Polo-like kinase 1 (PLK1) acts as a master regulator of mitosis and overexpresses in multiple tumor cells.The Cas9 and single guide sgRNA (sgRNA)-encoded plasmid was transduced to knockout Plk1 gene,which was expected to inhibit the expression of PLK1.Our studies demonstrated that ZEBRA enabled to transduce the CRISPR/Cas9 system with large size into the cells efficiently.The transduction with ZEBRA was cell line dependent,which showed~10-fold higher in CD44-positive cancer cell lines compared with CD44-negative ones.Furthermore,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 significantly.This 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

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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

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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 microenvironment.Owing to its importance in neuronal network formation,axon branching has been studied extensively during the past decades.It 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 understood.We 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

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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：哺乳动物细胞组织结构组装微尺度自底向上支架材料细胞生物学组织工程

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One-pot synthesis of N-doped petroleum coke-based microporous carbon for high-performance CO_(2) adsorption and supercapacitors

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Journal of Environmental Sciences 2024年第5期139卷 93-104页

作者： wenfu Zhu Yuqin Wang Fan Yao Xiaohong Wang Haoming zheng Guangzheng Ye Hairong Cheng Junliang Wu Haomin Huang Daiqi Ye School of Environment and Energy South China University of TechnologyGuangzhou 510006China National Engineering Laboratory for VOCs Pollution Control Technology and Equipment Guangzhou 510006China Guangdong Provincial Key Laboratory of Atmospheric Environment and Pollution Control(SCUT) Guangzhou 510006China Guangdong Provincial Engineering and Technology Research Centre for Environmental Risk Prevention and Emergency Disposal South China University of TechnologyGuangzhou 510006China

Waste resource utilization of petroleum coke is crucial for achieving global carbon emission reduction.Herein,a series of N-doped microporous carbons were fabricated from petroleum coke using a one-pot synthesis method.The as-prepared samples had a large specific surface area(up to 2512 m^(2)/g),a moderate-high N content(up to 4.82 at.%),and high population(55%)of ultra-micropores(<0.7 nm).Regulating the N content and ultra-microporosity led to efficient CO_(2)adsorption and separation.At ambient pressure,the optimal N-doped petroleum coke-based microporous carbon exhibited the highest CO_(2)uptake of 4.25 mmol/g at 25℃ and 6.57 mmol/g at 0℃.These values are comparable or even better than those of numerous previously reported adsorbents prepared by multistep synthesis,primarily due to the existence of ultra-micropores.The sample exhibited excellent CO_(2)/N_(2)selectivity at 25℃ owing to the abundant basic pyridinic and pyrrolic N species;and showed superior CO_(2)adsorption-desorption cycling performance,which was maintained at 97% after 10 cycles at 25℃.Moreover,petroleum coke-based microporous carbon,with a considerably high specific surface area and hierarchical pore structure,exhibited excellent electrochemical performance over the N-doped sample,maintaining a favorable specific capacitance of 233.25F/g at 0.5 A/g in 6 mol/L KOH aqueous electrolyte.This study provides insight into the influence of N-doping on the porous properties of petroleum coke-based carbon.Furthermore,the as-prepared carbons were found to be promising adsorbents for CO_(2)adsorption,CO_(2)/N_(2)separation and electrochemical application.

关键词： One-pot synthesis N-doped microporous carbon CO_(2)adsorption CO_(2)/N_(2)selectivity Electrochemical performance

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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 challenge.Nanoliposome-encapsulated light-sensitive compounds have excellent characteristics such as high temporal and spatial resolution,delayed drug clearance,and restricted drug biodistribution for neural modulation.In 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 weeks.For 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%,respectively.Simultaneously,the average power of local field potentials in the 0–15 Hz range degraded from 4.34 to 0.85 mW.In addition,the Nanolipo-Ru nanoparticles have the potential to provide more flexible timing of modulation than unencapsulated RuBi-GABA in the experiments.These results indicated that Nanolipo-Ru could be an effective platform for regulating neuronal electrophysiology.Furthermore,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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最优影响因素下超燃冲压发动机煤油流动水平管冷却

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机械设计与研究 2024年第2期40卷 29-31,39页

作者：芮雪郑少梅孙文福长治职业技术学院机械电子工程系山西长治046000 青岛理工大学机械与汽车工程学院山东青岛266109 三亚学院理工学院海南三亚572022

水和煤油在管道中的温度分布是不均匀的,且超燃冲压发动机煤油流动水平管冷却过程,没有针对不同区域换热过程进行进一步细分,导致煤油流动水平管冷却通道内速度场的分布规律不够精准,最优冷却因素分析结果失真,影响煤油的冷却效果。为... 详细信息

水和煤油在管道中的温度分布是不均匀的,且超燃冲压发动机煤油流动水平管冷却过程,没有针对不同区域换热过程进行进一步细分,导致煤油流动水平管冷却通道内速度场的分布规律不够精准,最优冷却因素分析结果失真,影响煤油的冷却效果。为解决这一问题,提出一种超燃冲压发动机煤油流动水平管最优冷却因素分析。通过构建航空煤油的对流换热控制模型,将整个换热过程分成不同的阶段,获取进口温度、压力、热流密度对传热的影响,计算出加热壁面内、壁面与流体三者间的传热系数,得出的流动水平管冷却的边界条件,并以此为依据获取对水平管内超临界航空煤油传热的影响因素,完成对超燃冲压发动机煤油流动水平管优化冷却。实验结果证明:所提方法的管壁温度拟合度与实际值基本一致,且冷却效果好,表明所提方法在超燃冲压发动机煤油流动水平管冷却方面具有一定的应用价值。

关键词：超燃冲压发动机煤油流动水平管冷却仿真方法局部高温分布规律

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A critical review of biochar-based nitrogen fertilizers and their effects on crop production and the environment

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Biochar 2022年第1期4卷 577-595页

作者： Yurong Gao zheng Fang Lukas Van Zwieten Nanthi Bolan Da Dong Bert F.Quin Jun Meng Fangbai Li Fengchang Wu Hailong Wang wenfu Chen Agronomy College Shenyang Agricultural UniversityShenyang 110866China Biochar Engineering Technology Research Center of Guangdong Province School of Environmental and Chemical EngineeringFoshan UniversityFoshan 528000China Key Laboratory of Biochar and Soil Improvement Ministry of Agriculture and Rural AffairsBeijingChina Department of Primary Industries New South Wales1243 Bruxner Highway2477 WollongbarNSWAustralia School of Agriculture and Environment The University of Western AustraliaPerthWA 6001Australia The UWA Institute of Agriculture The University of Western Australia6001 PerthWAAustralia School of Environmental and Resource Sciences Zhejiang A&F UniversityHangzhouZhejiang 311300China Quin Environmentals(NZ)Ltd PO Box 125122St.HeliersAuckland 1740New Zealand Guangdong Key Laboratory of Integrated Agro-Environmental Pollution Control and Management Institute of Eco-environmental and Soil SciencesGuangdong Academy of Sciences510650 GuangzhouChina State Key Laboratory of Environmental Criteria and Risk Assessment Chinese Research Academy of Environmental SciencesBeijing 100012China Guangdong Green Technologies Co. LtdFoshan 528100China

Globally,nitrogen(N)fertilizer demand is expected to reach 112 million tonnes to support food production for about 8 billion people.However,more than half of the N fertilizer is lost to the environment with impacts on air,water and soil quality,and biodiversity.Importantly,N loss to the environment contributes to greenhouse gas emissions and climate change.Nevertheless,where N fertilizer application is limited,severe depletion of soil fertility has become a major constraint to sustainable agriculture.To address the issues of low fertilizer N use efficiency(NUE),biochar-based N fertilizers(BBNFs)have been developed to reduce off-site loss and maximize crop N uptake.These products are generally made through physical mixing of biochar and N fertilizer or via coating chemical N fertilizers such as prilled urea with biochar.This review aims to describe the manufacturing processes of BBNFs,and to critically assess the effects of the products on soil properties,crop yield and N loss pathways.

关键词： Nitrogen use efficiency Controlled-release Retention Coating Biochar removing contaminants from

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Organs on microfluidic chips:a mini review

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Science China Chemistry 2014年第3期57卷 356-364页

作者： JIANG Bo zheng wenfu ZHANG Wei JIANG XingYu CAS Key Laboratory for Biological Effects of Nanomaterials and Nanosafety National Center for Nanoscience and Technology Graduate School of the Chinese Academy of Sciences

Microfluidic technology provides opportunities to create in vitro models with physiological microenvironment for cell study.Introducing the identified key aspects,including tissue-tissue interfaces,spatiotemporal chemical gradients,and dynamic mechanical forces,of living organs into the microfluidic system,"organs-on-chips"display an unprecedented application potential in a lot of biological fields such as fundamental physiological and pathophysiological research,drug efficacy and toxicity testing,and clinical diagnosis.Here,we review the recent development of organs-on-chips and briefly discuss their future challenges.

关键词：微流控芯片器官迷你微流体系统流控技术生物领域临床诊断毒性测试

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Effects of Early-stage Phased Rehabilitation Training on Acute Respiratory Distress Syndrome:A Systematic Review and Meta-analysis

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Medicinal Plant 2023年第5期14卷 72-75页

作者： wenfu ZHANG Chenyang zheng Feng ZHAO Department of Pediatric Rehabilitation Rehabilitation Medicine CenterTaihe HospitalHubei University of MedicineShiyan 442000China Department of Hydrotherapy and Physiotherapy Rehabilitation Medicine CenterTaihe HospitalHubei University of MedicineShiyan 442000China

[Objectives]To systematically evaluate the effects of early-stage phased rehabilitation training on the oxygenation index,ICU length of stay,duration of mechanical ventilation,and occurrence of complications(ventilator-associated pneumonia,pressure ulcers,delirium)in ARDS patients,thus contributing evidence for the clinical application of early-stage phased rehabilitation training.[Methods]The China National Knowledge Infrastructure(CNKI),Wanfang,and other databases were searched.Literature screening,data extraction,and systematic analysis of the included studies were performed using Revman software.[Results]Thirteen randomized controlled trials involving a total of 860 patients were included in this review.The results of the meta-analysis showed that compared to the traditional rehabilitation training group,the early-stage phased rehabilitation training group demonstrated a significant increase in the oxygenation index of ARDS patients[SMD=1.18,95%CI(1.01,1.35),P<0.01],with statistically significant differences.Furthermore,there were significant reductions in ICU length of stay[SMD=-0.70,95%CI(-0.90,-0.50),P<0.01],duration of mechanical ventilation[SMD=-1.15,95%CI(-1.36,-0.94),P<0.01],and occurrence of complications[OR=0.16,95%CI(0.10,0.26),P<0.01],all of which were statistically significant.[Conclusions]Early-stage phased pulmonary rehabilitation training for ARDS patients effectively improves the oxygenation index,shortens ICU length of stay and duration of mechanical ventilation,and reduces complications.These findings support the clinical application and promotion of early-stage phased rehabilitation training.

关键词： Early-stage phased rehabilitation training Acute respiratory distress syndrome(ARDS) Oxygenation index Systematic review

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