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Science China(Life Sciences) 2000年第4期43卷 433-441页

作者：王维刚张胜华李毅徐琳娜周京华甄永苏 Department of Oncology Institute of Medicinal Biotechnology Chinese Academy of Medical Sciences and Peking Union Medical College Beijing 100050 China

It was documented that type IV collagenase with two subtypes of 72 ku/MMP-2 and 92 ku/MMP-9 plays an important role in tumor invasion and metastasis. The endoplasmic reticulum (ER)- retained, single chain Fv antibody fragment (scFv) was used to inhibit the function of type IV collagenase. For expression in mammalian cells, the assembled scFv M97 gene with ER retention signal encoding 6 additional amino acids (SEKDEL) was reamplified by PCR. The amplified fragments were cloned into the pcDNA3.1 vector. The resulting plasmid was sequenced and then introduced into PG cells, a highly metastatic human lung cancer cell line, by lipofectAMINE method. The result of intrabody gene therapy showed that type IV collegenase expression was down regulated significantly as measured by ELISA. The biological behavior of PG cell, such as the ability of in vitro invasion through Matrigel, colony formation on soft agar, was also inhibited by scFv M97 transfection. Animal experiments in a xenograft model of human lung cancer showed that scFv M97 transfection significantly prolonged the survival time of nude mice. The results indicate that intracellular antibody technology represents a novel and efficient way to abrogate selectively the activity of type IV collagenase.

关键词： intracellular antibody (intrabody) type IV collagenase cancer invasion and metastasis gene therapy.

intracellular NHX-Type Cation/H＋ Antiporters inPlants

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Molecular Plant 2014年第2期7卷 261-263页

作者： Maria Reguera Elias Bassil Eduardo Blumwald Department of Plant Sciences Mail Stop 5 University of California One Shields Ave Davis CA 95616 USA

Cells depend on the homeostatic maintenance of pHwithin specific cellular compartments to ensure optimalconditions for metabolic and enzymatic processes as wellas protein structure and function. In the animal secre-tory pathway, cells maintain distinct luminal pHs withinvarious compartments （Paroutis et al., 2004）. Among themany molecular players that contribute to pH and ionhomeostasis in plants, Na＋（K＋）/H＋ exchangers （also knownas NHX-type cation/H＋ antiporters） appear to be particu-larly important for the regulation of a wide variety ofphysiological processes, including cell expansion, cellvolume regulation, osmotic adjustment, pH regulation,membrane trafficking, protein processing, and cellularstress responses （Pardo et al., 2006; Rodriguez-Rosaleset al., 2009; Bassil et al., 2012）. In plants, NHX antiportersappeared early in evolution and are ubiquitously encodedmembers of the CPA1 cation/H＋ antiporters subgroupthat belongs to the large family of monovalent cation/H＋ transporters CPA （Brett et al., 2005）. NHX antiport-ers are found, thus far, in all sequenced plant genomes（Bassil et al., 2012; Chanroj et al., 2012）. In Arabidopsis,the NHX family consists of eight isoforms, six of whichare intracellular （AtNHXl-AtNHX6）, located either to thevacuole （AtNHXl to AtNHX4） or endosomes （AtNHX5 andAtNHX6） and an additional two more divergent members（AtNHX7/SOSl and AtNHX8） at the plasma membrane（Bassil et al., 2012）. Orthologous sequences in each of thethree classes （plasma membrane, vacuolar, or endosomal）appear in all sequenced genomes, suggesting that distinctfunctional NHX classes appeared early in evolution andmay have conserved roles that are compartment-specific（Bassil et al., 2012）. Emerging new evidence highlightsthe importance of particular intracellular NHX antiport-ers in the regulation of vesicular and vacuolar pH andK＋ homeostasis. Vacuolar NHXs are needed to maintainK＋ homeostasis between the vacuole and cytosol, with-out which cell expansion is compromi

关键词： ion concentration plant genome Plasma membrane PH Regulation (biology) cell expansion intracellular antiporters Vacuoles Cellular processes

intracellular recording and morphology of tectal neurons activated by contralateral nucleus isthmi in toads

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Chinese Science Bulletin 1997年第19期42卷 1647-1651页

作者： Wang, SR Wu, GY Laboratory for Visual Information Processing Institute of Biophysics Chinese Academy of Sciences Beijing China

THE nucleus isthmi (NI) in lower vertebrates and its mammalian analogue, the nucleus para-bigeminalis, are all visual centres. They receive their tectal input ipsilaterally and project backipsilaterally in teleosts and birds, or bilaterally in amphibians and reptiles, as well as in mam-mals. It is well known that NI receives its visual excitation through the ipsilateral optic tec-tum (OT). NI mainly exerts inhibition on the ipsilateral OT in frogs.A recent study

关键词： nucleus isthmi tectum intracellular recording morphology toad

One-dimensional microstructure-assisted intradermal and intracellular delivery

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Bio-Design and Manufacturing 2019年第1期2卷 24-30页

作者： Wensen Jiang Liang Ma Xiaobin Xu Materials Science and Engineering School. Tongji University Shanghai 201804. China State Key Laboratory of Fluid Power and Mechatronic Systems. Zhejiang University. Hangzhou 310058. China School of Mechanical Engineering. Zhejiang University Hangzhou 310058 China

The advancement in the materials manufacturing at micrometer and nanometer scales has already enabled numerous applications in electronics, optics, chemistry, biology and medicine. Biomedical devices carrying micro-/nanostructures are currently being widely used in drug delivery, drug release, biosensing and therapy. New clinical methods for disease diagnosis and treatments are being developed enabled by nanotech no logy. One-dimensional (ID) structures are playi ng an importa nt role in the direct drug delivery both in vivo and ex vivo among various micro-/nanostructures. Here, in this paper, we reviewed recent progresses made on next-generation intradermal and intracellular delivery strategies and applications with focus on ID microstructure-based approaches.

关键词： Biomedical devices Intradermal intracellular ID nanostructure Drug delivery

intracellular chloride regulates the G_1/S cell cycle progression in gastric cancer cells

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World Journal of Gastrointestinal Oncology 2011年第8期3卷 119-122页

作者： Atsushi Shiozaki Eigo Otsuji Yoshinori Marunaka Division of Digestive Surgery Department of Surgery Kyoto Prefectural University of Medicine Kyoto 602-8566 Japan Department of Molecular Cell Physiology Graduate School of Medical Science Kyoto Prefectural University of Medicine Kyoto 602-8566 Japan Japan Institute for Food Education and Health St. Agnes' University Kyoto 602-8013 Japan

Recent studies show that ion channels/transporters play important roles in fundamental cellular functions. Several reports indicating the important roles of Cl- channels/transporters on cell proliferation suggest that the intracellular chloride concentration ([Cl-]i) regulated by them would be one of critical messengers. We investigated whether the [Cl-]i controls cell proliferation and cell cycle progression in human gastric cancer cells. Our studies indicated that furosemide, a blocker of Na+ /K+ /2Cl- cotransporter (NKCC), diminished cell growth by delaying the G1-S phase progression in gastric cancer cells with high expression and activity of NKCC. Furthermore, we found that the culture in the low Cl- medium (replacement of Cl- by NO3-) decreased the [Cl-]i and inhibited cell growth of gastric cancer cells and that this inhibition of cell growth was due to cell cycle arrest at the G0/G1 phase caused by diminutionof CDK2 and phosphorylated Rb. The culture of cells in the low Cl- medium significantly increased expressions of p21 mRNA and protein. In addition, the low Cl- medium induced phosphorylation of mitogen activated protein kinases (MAPKs). Treatment with an inhibitor of p38 or JNK significantly suppressed p21 upregulation caused by culture in a low Cl- medium and rescued gastric cancer cells from the low Cl- -induced G1 cell cycle arrest. These findings revealed that the [Cl-]i affects the cell proliferation via activation of MAPKs through upregulation of p21 in gastric cancer cells. Our results suggest that the [Cl-]i regulates important cellular functions in gastric cancer cells, leading to the development of novel therapeutic strategies.

关键词： intracellular chloride Cell proliferation Cell cycle Gastric cancer Cl- channel Cl- transporter

Preferential binding between intracellular organic matters and Al_(13) polymer to enhance coagulation performance

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Journal of Environmental Sciences 2019年第2期31卷 1-11页

作者： Ruiping Liu Tingting Guo Min Ma Mingquan Yan Jing Qi Chengzhi Hu Gang Liu Huijuan Liu Jiuhui Qu Walter van der Meer Key Laboratory of Drinking Water Science and Technology Research Center for Eco-Environmental SciencesChinese Academy of Sciences University of Chinese Academy of Sciences Beijing University of Technology Beijing Waterworks Group Department of Environmental Engineering Peking University Sanitary Engineering Department of Water ManagementFaculty of Civil Engineering and GeosciencesDelft University of Technology State Key Laboratory of Environmental Aquatic Chemistry Research Center for Eco-Environmental SciencesChinese Academy of Sciences Science and Technology University of Twente

Coagulation is the best available method for removing intracellular organic matter(IOM),which is released from algae cells and is an important precursor to disinfection by-products in drinking water treatment. To gain insight into the best strategy to optimize IOM removal, the coagulation performance of two Al salts, i.e., aluminum chloride(AlCl_3) and polyaluminum chloride(PACl, containing 81.2% Al_(13)), was investigated to illuminate the effect of Al species distribution on IOM removal. PACl showed better removal efficiency than AlCl_3 with regard to the removal of turbidity and dissolved organic carbon(DOC), owing to the higher charge neutralization effect and greater stability of pre-formed Al_(13) species. High pressure size exclusion chromatography analysis indicated that the superiority of PACl in DOC removal could be ascribed to the higher binding affinity between Al_(13) polymer and the low and medium molecular weight(MW) fractions of IOM. The results of differential log-transformed absorbance at 254 and 350 nm indicated more significant formation of complexes between AlCl_3 and IOM, which benefits the removal of tryptophan-like proteins thereafter. Additionally,PACl showed more significant superiority compared to AlCl_3 in the removal of <5 kD a and hydrophilic fractions, which are widely viewed as the most difficult to remove by *** study provides insight into the interactions between Al species and IOM, and advances the optimization of coagulation for the removal of IOM in eutrophic water.

关键词： intracellular organic matter Coagulation Polyaluminum chloride Molecular weight distribution Hydrophilic fraction

Effect of intracellular water release on hydro-mechanical behaviors of high kitchen waste content municipal solid waste

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Science China(Technological Sciences) 2019年第11期62卷 1907-1915页

作者： XU XiaoBing NI Da CHEN YunMin XU Hui YANG QinFeng XU SiFa Institute of Geotechnical Engineering Zhejiang University of TechnologyHangzhou 310023China MOE Key Laboratory of Soft Soils and Geoenvironmental Engineering Institute of Geotechnical EngineeringZhejiang UniversityHangzhou 310058China Department of Civil Engineering&Architecture Zhejiang Sci-Tech UniversityHangzhou 310018China The Architectural Design&Research Institute of Zhejiang University Co.Ltd Hangzhou 310028China

The release of intracellular water during degradation process contributes to the great leachate production and settlement of landfilled high kitchen waste content MSW(HKWC-MSW). An oven-drying and absorbent-paper combined method was proposed to measure the intracellular and interparticle water contents of HKWC-MSW. Two degradation experiments were carried out to study the release process of intracellular water and its effect on the hydro-mechanical behaviors of *** was found that the two degradation experiments showed similar degradation behaviors with BOD/COD decreasing with time in the early stage. In the first degradation experiment, most intracellular water was released during the first two months, and the degradation of degradable matter in kitchen waste(KW) was much slower than the release process of intracellular water. The particle size became smaller and the overall grain specific gravity increased during the fast release process of intracellular *** the second degradation experiment, after two-year degradation, the total leachate production was about 45.2% of the initial wet weight of HKWC-MSW specimen. Water retention capacity θ_f increased from 0.23 to 0.58 during 1–69 d, which might be caused by the decrease of particle size and compression of waste skeleton. As almost all the intracellular water was released after 80-day degradation, during the latter stage of leachate drainage under gravity, θ_f decreased and was close to the total volumetric water content. The total compression strain was about 0.39. The secondary compression strain during 1–80 d(i.e., about 0.07)was considered to be mainly resulted by the release of intracellular water and the subsequent drainage of leachate, and it accounted for about 22.6% of the total secondary compression strain.

关键词： municipal solid waste landfill degradation intracellular water particle size grain specific gravity water retention capacity settlement

Single-wavelength Excited Ratiometric Fluorescence pH Probe to Image intracellular Trafficking of Tobacco Mosaic Virus

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Chinese Journal of Polymer Science 2020年第6期38卷 587-592页

作者： Si-Jia Gao Zhuang Li Zhi-Cheng Sun Jin-Yue Wen Fu-Rong Li Xiao-Yang Du Yan Liu Ye Tian Zhong-Wei Niu Key Laboratory of Photochemical Conversion and Optoelectronic Materials Technical Institute of Physics and ChemistryChinese Academy of SciencesBeijing 100190China University of Chinese Academy of Sciences Beijing 100049China Beijing Engineering Research Centerof Printed Electronics Beijing Institute of Graphic CommunicationBeijing 102600China School of Future Technology University of Chinese Academy of SciencesBeijing 100049China

As a typical plant virus which has biocompatibility and high transfection efficiency,tobacco mosaic virus(TMV)has shown broad application potential in drug or gene delivery *** its intracellular trafficking is of great importance in investigation of its cytotoxicity,targeting site,and delivery efficiency,and is advantageous to designing new TMV-based drug delivery systems with different *** taking advantage of the regulated pH value of different organelles in a mammalian cell,we exploit a pH detection strategy to investigate the intracellular trafficking pathway of ***,we report a single-wavelength excited ratiometric fluorescent pH *** probe is constructed by simultaneously coupling pH-sensitive fluorescein isothiocyanate(FITC)and pH-insensitive rhodamine B isothiocyanate(RBIRC)onto the inner surface of *** fluorescence intensity ratio of FITC to RBITC excited at 488 nm responds specifically towards pH value over other interferential *** taking use of this single-wavelength excited ratiometric pH probe and confocal laser scanning microscopy,it is shown that the endocytosed TMV is located in a pH decreasing microenvironment and eventually enters *** work may provide important guidance on construction of TMV-based nano carriers.

关键词： Tobacco mosaic virus Ratiometric fluorescence pH probe intracellular trafficking

Spatiotemporal dynamics of cell abundance,colony size and intracellular toxin concentrations of pelagic and benthic Microcystis in Lake Caohai,China

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Journal of Environmental Sciences 2019年第10期31卷 184-196页

作者： Bing Feng Chunbo Wang Xingqiang Wu Cuicui Tian Meng Zhang Yingying Tian Bangding Xiao Key Laboratory of Algal Biology of Chinese Academy of Sciences Institute of Hydrobiology Chinese Academy of Sciences Wuhan 430072China Jiangxi Academy of Environmental Sciences Nanchang 330039 China Henan institute of technology Henan 453003 China

Lake Caohai has experienced extensive Microcystis blooms in recent years,and to improve its water quality,the local government carried out a series of water control *** better understand the dynamics of both pelagic and benthic Microcystis and their characteristics in Lake Caohai,we conducted a 1-year investigation from December 2015to December 2016 to gain a seasonal outlook on the distribution and dynamics of cell abundance,colony size and intracellular microcystins(MCs)of *** results indicated that the Microcystis bloom occupied primarily the northeastern region and then moved gradually from lakeshore to lake *** perennial southwesterly winds and the water inflow from northeast to southwest in Lake Caohai determined the spatiotemporal distribution of pelagic *** Microcystis was mainly distributed in the northeastern region in summer,occupied the lake center in autumn and then occupied the southeastern region in winter,determined by the sedimentation of pelagic Microcystis and the death of benthic *** colonies(20-60μm)overwintered more easily in both water column and *** concentrations of intracellular toxin of benthic Microcystis were observed to be significantly higher than those of pelagic *** might be because Microcystis synthesized large amount of MCs to acclimate to an unfavorable benthic *** knowledge on the dynamics of Microcystis expands our understanding of mechanisms underpinning the formation of Microcystis blooms.

关键词： Spatiotemporal dynamics Pelagic Microcystis Benthic Microcystis Cell abundance Colony size intracellular MCs

Visually intracellular Detection of Telomerase Activity Based on DNA Strand Displacement Reaction and Gold Nanoparticle Labeling

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Chinese Journal of Chemistry 2022年第6期40卷 693-698页

作者： Yajun Zhang Shen Yan Zhaoxin Chen Xin Jiang Shuang Rao Zhuoran Jiang Shanshan Qin Xiang Zhou Yuhao Du College of Chemistry and Molecular Sciences Key Laboratory of Biomedical Polymers of Ministry of EducationWuhan UniversityWuhanHubei 430072China

Telomerase,which is regarded as a common biomarker for early cancer diagnostics and a potential target for clinical therapies,has attracted considerable interests concerning its detection and ***,we propose a sensitive method by designing a gold nanoparticle(AuNP)probe for visually intracellular detection of telomerase *** AuNPs were functionalized with a telomer-ase substrate primer(SH-prime).A 6-carboxy-fluorescein(FAM)modified strand(FAM-probe)was attached to the surface of AuNP through its complementary stand(SH-attach).In the absence of telomerase,the fluorescence resonance energy transfer(FRET)from FAM to AuNPs results in efficient fluorescence *** the presence of telomerase,SH-primers on AuNPs were extended with the repeat units(TTAGGGG)*** extension sequence triggered the strand displacement of FAM-probe to restore the fluorescence *** is worth mentioning that the proposed strategy does not need to design complex hairpin structure and allows the meas-urement of telomerase in crude cell extracts down to 0.5 HeLa cells/μL in 2 *** addition,the present sensing platform can be ap-plied to the visually intracellular detection of telomerase activity in living cells.

关键词： Telomerase Nanoparticles DNA strand displacement intracellular FRET