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Chinese Chemical Letters 2023年第3期34卷 245-249页

作者： Xinqi Chang Nina Wang Depeng Jiang Hong-Yuan Chen Dechen Jiang State Key Laboratory of Analytical Chemistry for Life Science School of Chemistry and Chemical EngineeringNanjing UniversityNanjing 210093China Department of Respiratory Medicine The Second Affiliated HospitalChongqing Medical UniversityChongqing 400010China

Angiotensin-converting enzyme 2(ACE2) is not only an enzyme but also a functional receptor on cell membrane for severe acute respiratory syndrome coronavirus 2(SARS-CoV-2). Here, the activity of ACE2 in single living cell is firstly determined using a nanokit coupled electrospray ionization mass spectrometry(nanokit-ESI-MS). Upon the insertion of a micro-capillary into the living h ACE2-CHO cell and the electrochemical sorting of the cytosol, the target ACE2 enzyme hydrolyses angiotensin II inside the capillary to generate angiotensin 1-7. After the electrospray of the mixture at the tip of the capillary, the product is differentiated from the substrate in molecular weight to achieve the detection of ACE2 activity in single cells. The further measurement illustrates that the inflammatory state of cells does not lead to the significant change of ACE2 catalytic activity, which elucidates the relationship between intracellular ACE2 activity and inflammation at single cell level. The established strategy will provide a specific analytical method for further studying the role of ACE2 in the process of virus infection, and extend the application of nanokit based single cell analysis.

关键词： Nanokit Electrospray ionization mass spectrometry single cell analysis The enzymatic activity ACE2

analysis of Amino Acids in a single Human Red Blood cell by Capillary Zone Electrophoresis with Intracellular NDA- derivatization and Electrochemical Detection

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Chinese Chemical Letters 2002年第7期13卷 655-657页

作者： Qian DONG Xiao Lei WANG Lan Lan ZHU Wen Rui JIN School of Chemistry and Chemical Engineering Shandong UniversityJinan 250100

A novel method for determination of amino acids in individual human red blood cells has been developed. In this method, the derivatization reagents (NDA and CN-) are introduced into living cells by electroporation. After completion of derivatization, the amino acids in a single cell is determined by capillary zone electrophoresis with end-column amperometric detection.

关键词： Capillary electrophoresis electrochemical detection single cell analysis amino acid.

Universal cell and tissue acquisition/analysis system: from single cell adhesion measurement and collection to archival FFPE brain tissue microdissection

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Universal cell and tissue acquisition/analysis system: from ...

Microfluidic adhesion analysis of single glioma cells for evaluating the effect of drugs

中国神经科学学会第十二届全国学术会议

作者： Zhongcai MA Kristen DEIGNAN Anialac ZAVALA Christopher K WILLIAMS Harry V.VINTERS Lili C.KUDO Stanislav L.KARSTEN NeuroInDx Inc. Department of Pathology David Geffen School of Medicine at UCLA

Objective: analysis of specific cells from cultures or isolation of subanatomical regions from heterogeneous tissues is essential in today’s biomedical research. single cell adhesion properties are a key to understanding cellular properties in health and disease. Tissue microdissection from fixed tumor biopsies and archived FFPE samples is much needed yet technically challenging. Commercially available single cell technologies do not provide concurrent single cell adhesion force measurement and acquisition, and current microdissection platforms are sample specific, complex and expensive. Methods: We applied our vacuum impulse based cell and tissue acquisition technology for single cell adhesion force measurement and fixed tissue microdissection. With our recent platform, Unipic K+?, adhesion strength for individual cells from SH-SY5 Y, CHO, and 3 T3 cultures were measured based on capillary aspiration techniques. single cells were collected and dispensed immediately into individual wells for clonal expansion.. A novel DNA extraction method was developed to acquire region specific DNA from 5 to 20 μm thick PFA-fixed and FFPE archived human brain tissue sections using disposable capillary units preloaded with alkaline solution and a microscope stage heater set at 100°C. Results: The new system collects individual cells from adherent cultures in as small as 15 nl, compatible with various downstream single cell analyses. Manual and automatic extraction algorithms were developed for acquisition of tissue region of interest（ROI）. Dissociation and collection of ROIs down to 20 μm in diameter can be performed on fixed tissue sections within 30 s. for DNA isolation. Conclusion: A universal platform has been developed for fresh frozen and fixed tissue microdissection, single cell acquisition, adhesion force measurements and deposition into single wells for various downstream applications.

关键词： adhesion single cell analysis tissue microdissection archival tissue acquisition dispensing

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Science China Chemistry 2020年第6期63卷 865-870页

作者： Qiang Zhang Sifeng Mao Weiwei Li Qiushi Huang Shuo Feng Zhanying Hong Jin-Ming Lin Department of Chemistry Beijing Key Laboratory of Micronalytical Methods and InstrumentationMOE Key Laboratory of Bioorganic Phosphorus Chemistry&Chemical BiologyTsinghua UniversityBeijing 100084China Department of Pharmaceutical analysis School of PharmacyShanghai Key Laboratory for Pharmaceutical Metabolites ResearchNaval Medical UniversityShanghai 200433China

Cancer metastasis is one of the most serious problems for tumor therapy,which is closely related to cell adhesion and deadhesion *** comprehension of cell adhesion ability will benefit drug ***,a biomimetic microfluidic enzyme digestion method was proposed to gently measure the influence of drugs on cell-matrix adhesion ability at the single cell *** method can selectively digest the extracellular matrix(ECM)that linked to a single cell,and the trypsin concentration around the cell is relatively uniform and constant,thus the measured cell adhesion strength should be *** available anti-cancer agents including 5-fluorouracil(5-FU),actinomycin D(Act D),temozolomide(TMZ)and allicin were evaluated,and the data showed only TMZ and allicin can inhibit cell adhesion significantly under our experiment *** influence of TMZ became more and more obvious as the increase of duration and the effect became prominent only after 6 h adhesion process,which could provide a quick evaluation of whether the drugs are effective to cancer cell(compared with Calcein-AM/PI cell viability test).The adhesion strength of U87 cells decreased when the concentration of TMZ increased,and the effect of TMZ can be effectively inhibited by adding lactic acid to culture medium,which indicated acidic tumor microenvironment could promote drug resistance of tumor *** from conventional evaluation methods which focus on the drugs’influence on cellular viability or metabolism,this work provides a new perspective to study the effect of drugs,which is helpful to enrich the drug evaluation system.

关键词： microfluidic single cell analysis cell adhesion drug evaluation

2D profiling of tumor chemotactic and molecular phenotype at single cell resolution using a SERS-microfluidic chip

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Nano Research 2022年第5期15卷 4357-4365页

作者： Yizhi Zhang Lei Wu Kuo Yang Shenfei Zong Zhuyuan Wang Yiping Cui Advanced Photonics Center School of Electronic Science and EngineeringSoutheast UniversityNanjing 210096China Department of Biomedical Engineering Nanjing University of Aeronautics and AstronauticsNanjing 211106China

Emerging single-cell technologies create new opportunities for unraveling tumor ***,the development of high-content phenotyping platform is still at its ***,we develop a microfluidic chip for two-dimensional(2D)profiling of tumor chemotactic and molecular features at single cell *** cells were captured by the triangular micropillar arrays in the cell-loading channel,facilitating downstream single-cell *** 2D phenotyping,the chemotactic properties of tumor cells were visualized through cellular migratory behavior in microchannels,while their protein expression was profiled with multiplex surface enhanced Raman scattering(SERS)nanovectors,in which Raman reporter-embedded gold@silver core-shell nanoparticles(Au@Ag REPs)were modified with DNA aptamers targeting cellular surface *** a proof of concept,breast cancer cells with diverse phenotypes were tested on the chip,demonstrating the capability of this platform for simultaneous chemotactic and molecular *** chip is expected to provide a powerful tool for investigating tumor heterogeneity and promoting clinical precision medicine.

关键词： gold@silver nanoparticles surface enhanced Raman spectroscopy microfluidic chip single cell analysis two-dimensional(2D)phenotype profiling

Determination of Amino Acids in an Individual Erythrocyteby Capillary Electrophoresis with Intracellular FITC-derivatization and Laser-induced Fluorescence Detection

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Chinese Chemical Letters 2003年第9期14卷 952-954页

作者： HuaZHANG WenRuiJIN StateKeyLaboratoryofMicrobialTechnology ShandongUniversityJinan250100 SchoolofChemistry-andChemicalEngineering ShandongUniversityJinan250100

A novel approach for analysis of amino acids in individual erythrocytes was established. In this method, the derivatization reagent was introduced into the living cells by electroporation. After derivatization, the am... 详细信息

关键词： Capillary electrophoresis laser-induced fluorescence detection single cell analysis amino acid.

Transient induction of actin cytoskeletal remodeling associated with dedifferentiation,proliferation,and redifferentiation stimulates cardiac regeneration

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Acta Pharmaceutica Sinica B 2024年第6期14卷 2537-2553页

作者： Wenbin Fu Qiao Liao Yu Shi Wujian Liu Hongmei Ren Chunmei Xu Chunyu Zeng Department of Cardiology Daping Hospitalthe Third Military Medical University(Army Medical University)Chongqing 400042China Key Laboratory of Geriatric Cardiovascular and Cerebrovascular Disease Research Ministry of Education of ChinaChongqing 400042China Chongqing Key Laboratory for Hypertension Research Chongqing Cardiovascular Clinical Research CenterChongqing Institute of CardiologyChongqing 400042China State Key Laboratory of Trauma Burns and Combined InjuryDaping Hospitalthe Third Military Medical UniversityChongqing 400042China Cardiovascular Research Center Chongqing CollegeUniversity of Chinese Academy of SciencesChongqing 400042China State Key Laboratory of Trauma and Chemical Poisoning Chongqing 400042China

The formation of new and functional cardiomyocytes requires a 3-step process:dedifferentiation,proliferation,and redifferentiation,but the critical genes required for efficient dedifferentiation,proliferation,and redifferentiation remain *** our study,a circular trajectory using single-nucleus RNA sequencing of the pericentriolar material 1 positive(PCM1^(+))cardiomyocyte nuclei from hearts 1 and 3 days after surgery-induced myocardial infarction(MI)on postnatal Day 1 was reconstructed and demonstrated that actin remodeling contributed to the dedifferentiation,proliferation,and redifferentiation of cardiomyocytes after *** identified four top actin-remodeling regulators,namely Tmsb4x,Tmsb10,Dmd,and Ctnna3,which we collectively referred to as *** expressed changes of 2D2P,using a polycistronic non-integrating lentivirus driven by Tnnt2(cardiac-specific troponin T)promoters(Tnnt2-2D2P-NIL),efficiently induced transiently proliferative activation and actin remodeling in postnatal Day 7 cardiomyocytes and adult ***,the intramyocardial delivery of Tnnt2-2D2P-NIL resulted in a sustained improvement in cardiac function without ventricular dilatation,thickened septum,or fatal arrhythmia for at least 4 *** conclusion,this study highlights the importance of actin remodeling in cardiac regeneration and provides a foundation for new gene-cocktail-therapy approaches to improve cardiac repair and treat heart failure using a novel transient and cardiomyocyte-specific viral construct.

关键词： single cell analysis Actin remodeling Tmsb4x Tmsb10 Dmd Ctnna3 Myocardial infarction Cardiomyocytes proliferation Cardiac regeneration Genetic therapy

Cross-regulation between CDK and MAPK control cellular fate

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Quantitative Biology 2021年第3期9卷 341-359页

作者： Eric Durandau Serge Peiet Department of Fundamental Microbiology University of LausanneCH-1015 LausanneSwitzerland

Background:Commitment to a new cell cycle is controlled by a number of cellular ***-activated protein kinase(MAPK)pathways,which transduce multiple extracellular cues,have been shown to be interconnected with the cell cycle and can modulate its ***:In budding yeast,we have introduced fluorescent biosensors that monitor in real time the signaling activity of the MAPKs Fus3 and Kssl and the cyclin-dependent kinase(CDK)in individual *** have quantified in hundreds of live single cells the interplay between the MAPKs regulating the mating response and the CDK controlling cell cycle ***:Different patterns of MAPK activity dynamics could be identified by clustering cells based on their CDK activity,denoting the tight relationship between these two cellular *** data suggest that beyond the already well-established mechanisms of regulation between the MAPK and the CDK,additional mechanisms remain to be ***:A tight interplay between MAPK pathways and the cell cycle is essential to control cellular proliferation and cell fate decisions.

关键词： MAPK signaling cell cycle yeast mating single cell analysis fluorescent biosensors

Recent updates on the biological basis of heterogeneity in bone marrow stromal cells/ skeletal stem cells

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Biomaterials Translational 2022年第1期3卷 3-16页

作者： Deepika Arora Pamela Gehron Robey National Institute of Dental and Craniofacial Research National Institutes of HealthDepartment of Health and Human ServicesBethesdaMDUSA Biosystems and Biomaterials Division National Institute of Standards and TechnologyDepartment of CommerceGaithersburgMDUSA Department of Biotechnology School of Biological Engineering&Life SciencesShobhit Institute of Engineering&Technology(Deemed-to-be-University)MeerutIndia

Based on studies over the last several decades,the self-renewing skeletal lineages derived from bone marrow stroma could be an ideal source for skeletal tissue ***,the markers for osteogenic precursors;i.e.,bone marrow-derived skeletal stem cells(SSCs),in association with other cells of the marrow stroma(bone marrow stromal cells,BMSCs)and their heterogeneous nature both in vivo and in vitro remain to be *** review aims to highlight:i)the importance of distinguishing BMSCs/SSCs from other“mesenchymal stem/stromal cells”,and ii)factors that are responsible for their heterogeneity,and how these factors impact on the differentiation potential of SSCs towards *** prospective role of SSC enrichment,their expansion and its impact on SSC phenotype is *** has also been given to emerging single cell RNA sequencing approaches in scrutinizing the unique population of SSCs within the BMSC population,along with their committed *** the factors involved in heterogeneity may help researchers to improvise their strategies to isolate,characterize and adopt best culture practices and source identification to develop standard operating protocols for developing reproducible stem cells ***,more scientific understanding of the molecular basis of heterogeneity is warranted that may be obtained from the robust high-throughput functional transcriptomics of single cells or clonal populations.

关键词： bone marrow stromal cells clonal analysis heterogeneity single cell analysis skeletal stem cells