arXiv - QuanBio - Cell Behavior最新文献

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Persistent pseudopod splitting is an effective chemotaxis strategy in shallow gradients 持续的伪足分裂是浅梯度中一种有效的趋化策略
arXiv - QuanBio - Cell Behavior Pub Date : 2024-09-14 DOI: arxiv-2409.09342
Albert Alonso, Julius B. Kirkegaard, Robert G. Endres
{"title":"Persistent pseudopod splitting is an effective chemotaxis strategy in shallow gradients","authors":"Albert Alonso, Julius B. Kirkegaard, Robert G. Endres","doi":"arxiv-2409.09342","DOIUrl":"https://doi.org/arxiv-2409.09342","url":null,"abstract":"Single-cell organisms and various cell types use a range of motility modes\u0000when following a chemical gradient, but it is unclear which mode is best suited\u0000for different gradients. Here, we model directional decision-making in\u0000chemotactic amoeboid cells as a stimulus-dependent actin recruitment contest.\u0000Pseudopods extending from the cell body compete for a finite actin pool to push\u0000the cell in their direction until one pseudopod wins and determines the\u0000direction of movement. Our minimal model provides a quantitative understanding\u0000of the strategies cells use to reach the physical limit of accurate chemotaxis,\u0000aligning with data without explicit gradient sensing or cellular memory for\u0000persistence. To generalize our model, we employ reinforcement learning\u0000optimization to study the effect of pseudopod suppression, a simple but\u0000effective cellular algorithm by which cells can suppress possible directions of\u0000movement. Different pseudopod-based chemotaxis strategies emerge naturally\u0000depending on the environment and its dynamics. For instance, in static\u0000gradients, cells can react faster at the cost of pseudopod accuracy, which is\u0000particularly useful in noisy, shallow gradients where it paradoxically\u0000increases chemotactic accuracy. In contrast, in dynamics gradients, cells form\u0000textit{de novo} pseudopods. Overall, our work demonstrates mechanical\u0000intelligence for high chemotaxis performance with minimal cellular regulation.","PeriodicalId":501321,"journal":{"name":"arXiv - QuanBio - Cell Behavior","volume":"38 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142259007","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Geometric Effects in Large Scale Intracellular Flows 大规模细胞内流动的几何效应
arXiv - QuanBio - Cell Behavior Pub Date : 2024-09-10 DOI: arxiv-2409.06763
Olenka Jain, Brato Chakrabarti, Reza Farhadifar, Elizabeth R. Gavis, Michael J. Shelley, Stanislav Y. Shvartsman
{"title":"Geometric Effects in Large Scale Intracellular Flows","authors":"Olenka Jain, Brato Chakrabarti, Reza Farhadifar, Elizabeth R. Gavis, Michael J. Shelley, Stanislav Y. Shvartsman","doi":"arxiv-2409.06763","DOIUrl":"https://doi.org/arxiv-2409.06763","url":null,"abstract":"This work probes the role of cell geometry in orienting self-organized fluid\u0000flows in the late stage Drosophila oocyte. Recent theoretical work has shown\u0000that a model, which relies only on hydrodynamic interactions of flexible,\u0000cortically anchored microtubules (MTs) and the mechanical loads from molecular\u0000motors moving upon them, is sufficient to generate observed flows. While the\u0000emergence of flows has been studied in spheres, oocytes change shape during\u0000streaming and it was unclear how robust these flows are to the geometry of the\u0000cell. Here we use biophysical theory and computational analysis to investigate\u0000the role of geometry and find that the axis of rotation is set by the shape of\u0000the domain and that the flow is robust to biologically relevant perturbations\u0000of the domain shape. Using live imaging and 3D flow reconstruction, we test the\u0000predictions of the theory/simulation, finding consistency between the model and\u0000live experiments, further demonstrating a geometric dependence on flow\u0000direction in late-stage Drosophila oocytes.","PeriodicalId":501321,"journal":{"name":"arXiv - QuanBio - Cell Behavior","volume":"148 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142182177","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Motion Ordering in Cellular Polar-polar and Polar-nonpolar Interactions 细胞极性-极性和极性-非极性相互作用中的运动排序
arXiv - QuanBio - Cell Behavior Pub Date : 2024-09-09 DOI: arxiv-2409.05333
Katsuyoshi Matsushita, Taiko Arakaki, Koichi Fujimoto
{"title":"Motion Ordering in Cellular Polar-polar and Polar-nonpolar Interactions","authors":"Katsuyoshi Matsushita, Taiko Arakaki, Koichi Fujimoto","doi":"arxiv-2409.05333","DOIUrl":"https://doi.org/arxiv-2409.05333","url":null,"abstract":"We examine the difference in motion ordering between cellular systems with\u0000and without information transfer to evaluate the effect of the polar--polar\u0000interaction through mutual guiding, which enables cells to inform other cells\u0000of their moving directions. We compare this interaction with the\u0000polar--nonpolar interaction through cell motion triggered by cellular contact,\u0000which cannot provide information on the moving directions. We model these\u0000interactions on the basis of the cellular Potts model. We calculate the order\u0000parameter of the polar direction in the interactions and examine the cell\u0000concentration and surface tension conditions of ordering. The results suggest\u0000that the polar--polar interaction through mutual guiding efficiently induces\u0000the motion ordering in comparison with the polar-nonpolar interaction for\u0000contact triggering, except in cases of weak driving. The results also show that\u0000the polar--polar interaction efficiently accelerates the collective motion\u0000compared with the polar--nonpolar interaction.","PeriodicalId":501321,"journal":{"name":"arXiv - QuanBio - Cell Behavior","volume":"49 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142182176","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Modelling how lamellipodia-driven cells maintain persistent migration and interact with external barriers 模拟叶状薄片驱动的细胞如何保持持续迁移并与外部障碍相互作用
arXiv - QuanBio - Cell Behavior Pub Date : 2024-09-07 DOI: arxiv-2409.04772
Shubhadeep Sadhukhan, Cristina Martinez-Torres, Samo Penič, Carsten Beta, Aleš Iglič, Nir S Gov
{"title":"Modelling how lamellipodia-driven cells maintain persistent migration and interact with external barriers","authors":"Shubhadeep Sadhukhan, Cristina Martinez-Torres, Samo Penič, Carsten Beta, Aleš Iglič, Nir S Gov","doi":"arxiv-2409.04772","DOIUrl":"https://doi.org/arxiv-2409.04772","url":null,"abstract":"Cell motility is fundamental to many biological processes, and cells exhibit\u0000a variety of migration patterns. Many motile cell types follow a universal law\u0000that connects their speed and persistency, a property that can originate from\u0000the intracellular transport of polarity cues due to the global actin retrograde\u0000flow. This mechanism was termed the ``Universal Coupling between cell Speed and\u0000Persistency\"(UCSP). Here we implemented a simplified version of the UCSP\u0000mechanism in a coarse-grained ``minimal-cell\" model, which is composed of a\u0000three-dimensional vesicle that contains curved active proteins. This model\u0000spontaneously forms a lamellipodia-like motile cell shape, which is however\u0000sensitive and can depolarize into a non-motile form due to random fluctuations\u0000or when interacting with external obstacles. The UCSP implementation introduces\u0000long-range inhibition, which stabilizes the motile phenotype. This allows our\u0000model to describe the robust polarity observed in cells and explain a large\u0000variety of cellular dynamics, such as the relation between cell speed and\u0000aspect ratio, cell-barrier scattering, and cellular oscillations in different\u0000types of geometric confinements.","PeriodicalId":501321,"journal":{"name":"arXiv - QuanBio - Cell Behavior","volume":"116 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142182178","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Synchronized Memory-Dependent Intracellular Oscillations for a Cell-Bulk ODE-PDE Model in $mathbb{R}^2$ $mathbb{R}^2$ 中细胞群 ODE-PDE 模型的同步记忆依赖性胞内振荡
arXiv - QuanBio - Cell Behavior Pub Date : 2024-09-01 DOI: arxiv-2409.00623
Merlin Pelz, Michael J. Ward
{"title":"Synchronized Memory-Dependent Intracellular Oscillations for a Cell-Bulk ODE-PDE Model in $mathbb{R}^2$","authors":"Merlin Pelz, Michael J. Ward","doi":"arxiv-2409.00623","DOIUrl":"https://doi.org/arxiv-2409.00623","url":null,"abstract":"For a cell-bulk ODE-PDE model in $mathbb{R}^2$, a hybrid\u0000asymptotic-numerical theory is developed to provide a new theoretical and\u0000computationally efficient approach for studying how oscillatory dynamics\u0000associated with spatially segregated dynamically active ``units\" or ``cells\"\u0000are regulated by a PDE bulk diffusion field that is both produced and absorbed\u0000by the entire cell population. The study of oscillator synchronization in a PDE\u0000diffusion field was one of the initial aims of Yoshiki Kuramoto's foundational\u0000work. For this cell-bulk model, strong localized perturbation theory, as\u0000extended to a time-dependent setting, is used to derive a new\u0000integro-differential ODE system that characterizes intracellular dynamics in a\u0000memory-dependent bulk-diffusion field. For this nonlocal reduced system, a\u0000novel fast time-marching scheme, relying in part on the\u0000emph{sum-of-exponentials method} to numerically treat convolution integrals,\u0000is developed to rapidly and accurately compute numerical solutions to the\u0000integro-differential system over long time intervals. For the special case of\u0000Sel'kov reaction kinetics, a wide variety of large-scale oscillatory dynamical\u0000behavior including phase synchronization, mixed-mode oscillations, and\u0000quorum-sensing are illustrated for various ranges of the influx and efflux\u0000permeability parameters, the bulk degradation rate and bulk diffusivity, and\u0000the specific spatial configuration of cells. Results from our fast algorithm,\u0000obtained in under one minute of CPU time on a laptop, are benchmarked against\u0000PDE simulations of the cell-bulk model, which are performed with a commercial\u0000PDE solver, that have run-times that are orders of magnitude larger.","PeriodicalId":501321,"journal":{"name":"arXiv - QuanBio - Cell Behavior","volume":"10 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142182179","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
A kinetic chemotaxis model and its diffusion limit in slab geometry 板块几何中的动力学趋化模型及其扩散极限
arXiv - QuanBio - Cell Behavior Pub Date : 2024-08-30 DOI: arxiv-2408.17243
Herbert Egger, Kathrin Hellmuth, Nora Philippi, Matthias Schlottbom
{"title":"A kinetic chemotaxis model and its diffusion limit in slab geometry","authors":"Herbert Egger, Kathrin Hellmuth, Nora Philippi, Matthias Schlottbom","doi":"arxiv-2408.17243","DOIUrl":"https://doi.org/arxiv-2408.17243","url":null,"abstract":"Chemotaxis describes the intricate interplay of cellular motion in response\u0000to a chemical signal. We here consider the case of slab geometry which models\u0000chemotactic motion between two infinite membranes. Like previous works, we are\u0000particularly interested in the asymptotic regime of high tumbling rates. We\u0000establish local existence and uniqueness of solutions to the kinetic equation\u0000and show their convergence towards solutions of a parabolic Keller-Segel model\u0000in the asymptotic limit. In addition, we prove convergence rates with respect\u0000to the asymptotic parameter under additional regularity assumptions on the\u0000problem data. Particular difficulties in our analysis are caused by vanishing\u0000velocities in the kinetic model as well as the occurrence of boundary terms.","PeriodicalId":501321,"journal":{"name":"arXiv - QuanBio - Cell Behavior","volume":"50 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142182184","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Graph polyhedral divisions in growing cell aggregates 生长细胞聚集体中的图形多面体分裂
arXiv - QuanBio - Cell Behavior Pub Date : 2024-08-14 DOI: arxiv-2408.07551
Urban Železnik, Matej Krajnc, Tanmoy Sarkar
{"title":"Graph polyhedral divisions in growing cell aggregates","authors":"Urban Železnik, Matej Krajnc, Tanmoy Sarkar","doi":"arxiv-2408.07551","DOIUrl":"https://doi.org/arxiv-2408.07551","url":null,"abstract":"In the recently proposed Graph Vertex Model (GVM), cellular rearrangements\u0000are implemented as local graph transformations of the cell aggregate,\u0000represented by a knowledge graph [1]. This study extends GVM to incorporate\u0000cell division, a critical biological process involved in morphogenesis,\u0000homeostasis, and disease progression. Like cellular rearrangements, cell\u0000division in GVM begins by identifying a subgraph of nodes and links, involved\u0000in the division, by matching suitable graph patterns or templates within the\u0000full knowledge graph. The matched subgraph is then transformed to incorporate\u0000topological changes within the knowledge graph, caused by the division event.\u0000Importantly, when this transformation is applied to a polygon in a 2D tiling,\u0000it performs the transformation, required to divide a polygon, indicating that\u0000the 3D graph transformation is general and applicable also to 2D vertex models.\u0000Our extension of GVM enables the study of the dynamics of growing cell\u0000aggregates in 3D to offer new insights into developmental processes and cancer\u0000growth.","PeriodicalId":501321,"journal":{"name":"arXiv - QuanBio - Cell Behavior","volume":"55 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142182180","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Nonthermal driving forces in cells revealed by nonequilibrium fluctuations 非平衡波动揭示细胞中的非热驱动力
arXiv - QuanBio - Cell Behavior Pub Date : 2024-08-13 DOI: arxiv-2408.06683
Yuika Ueda, Shinji Deguchi
{"title":"Nonthermal driving forces in cells revealed by nonequilibrium fluctuations","authors":"Yuika Ueda, Shinji Deguchi","doi":"arxiv-2408.06683","DOIUrl":"https://doi.org/arxiv-2408.06683","url":null,"abstract":"The mechanical properties within living cells play a critical role in the\u0000adaptive regulation of their biological functions upon environmental and\u0000internal stimuli. While these properties exhibit nonequilibrium dynamics due to\u0000the thermal and nonthermal forces that universally coexist in\u0000actin-myosin-active proliferative cells, quantifying them within such complex\u0000systems remains challenging. Here, we develop a nonequilibrium framework that\u0000combines fluorescence correlation spectroscopy (FCS) measurements of\u0000intracellular diffusion with nonequilibrium theory to quantitatively analyze\u0000cell-specific nonthermal driving forces and cellular adaptability. Our results\u0000reveal that intracellular particle diffusion is influenced not only by common\u0000thermal forces but also by nonthermal forces generated by approximately 10-100\u0000motor proteins. Furthermore, we derive a physical parameter that quantitatively\u0000assesses the sensitivity of intracellular particle responses to these\u0000nonthermal forces, showing that systems with more active diffusion exhibit\u0000higher response sensitivity. Our work highlights the biological fluctuations\u0000arising from multiple interacting elements, advancing the understanding of the\u0000complex mechanical properties within living cells.","PeriodicalId":501321,"journal":{"name":"arXiv - QuanBio - Cell Behavior","volume":"22 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142182181","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Stochastic Gene Expression Model of Nuclear-to-Cell Ratio Homeostasis 核细胞比平衡的随机基因表达模型
arXiv - QuanBio - Cell Behavior Pub Date : 2024-07-26 DOI: arxiv-2407.19066
Xuesong Bai, Thomas G. Fai
{"title":"Stochastic Gene Expression Model of Nuclear-to-Cell Ratio Homeostasis","authors":"Xuesong Bai, Thomas G. Fai","doi":"arxiv-2407.19066","DOIUrl":"https://doi.org/arxiv-2407.19066","url":null,"abstract":"Cell size varies between different cell types, and between different growth\u0000and osmotic conditions. However, the nuclear-to-cell volume ratio (N/C ratio)\u0000remains nearly constant. In this paper, we build on existing deterministic\u0000models of N/C ratio homeostasis and develop a simplified gene translation model\u0000to study the effect of stochasticity on the N/C ratio homeostasis. We solve the\u0000corresponding chemical master equation and obtain the mean and variance of the\u0000N/C ratio. We also use a Taylor expansion approximation to study the effects of\u0000the system size on the fluctuations of the N/C ratio. We then combine the\u0000translation model with a cell division model to study the effects of extrinsic\u0000noises from cell division on the N/C ratio. Our model demonstrates that the N/C\u0000ratio homeostasis is maintained when the stochasticity in cell growth is taken\u0000into account, that the N/C ratio is largely determined by the gene fraction of\u0000nuclear proteins, and that the fluctuations in the N/C ratio diminish as the\u0000system size increases.","PeriodicalId":501321,"journal":{"name":"arXiv - QuanBio - Cell Behavior","volume":"11 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-07-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141863942","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Learning a phenomenological theory for contact-interactions between motile cells from collision experiments 从碰撞实验中学习运动细胞间接触互动的现象学理论
arXiv - QuanBio - Cell Behavior Pub Date : 2024-07-24 DOI: arxiv-2407.17268
Tom Brandstätter, Emily Brieger, David B. Brückner, Georg Ladurner, Joachim Rädler, Chase P. Broedersz
{"title":"Learning a phenomenological theory for contact-interactions between motile cells from collision experiments","authors":"Tom Brandstätter, Emily Brieger, David B. Brückner, Georg Ladurner, Joachim Rädler, Chase P. Broedersz","doi":"arxiv-2407.17268","DOIUrl":"https://doi.org/arxiv-2407.17268","url":null,"abstract":"The migration behavior of colliding cells is critically determined by\u0000transient contact-interactions. During these interactions, the motility\u0000machinery, including the front-rear polarization of the cell, dynamically\u0000responds to surface protein-mediated transmission of forces and biochemical\u0000signals between cells. While biomolecular details of such contact-interactions\u0000are increasingly well understood, it remains unclear what biophysical\u0000interaction mechanisms govern the cell-level dynamics of colliding cells and\u0000how these mechanisms vary across cell types. Here, we develop a\u0000phenomenological theory based on eleven candidate contact-interaction\u0000mechanisms coupling cell position, shape, and polarity. Using high-throughput\u0000micropattern experiments, we detect which of these phenomenological\u0000contact-interactions captures the interaction behaviors of cells. We find that\u0000various cell types - ranging from mesenchymal to epithelial cells - are\u0000accurately captured by a single model with only two interaction mechanisms:\u0000polarity-protrusion coupling and polarity-polarity coupling. The qualitatively\u0000different interaction behaviors of distinct cells, as well as cells subject to\u0000molecular perturbations of surface protein-mediated signaling, can all be\u0000quantitatively captured by varying the strength and sign of the\u0000polarity-polarity coupling mechanism. Altogether, our data-driven\u0000phenomenological theory of cell-cell interactions reveals polarity-polarity\u0000coupling as a versatile and general contact-interaction mechanism, which may\u0000underlie diverse collective migration behavior of motile cells.","PeriodicalId":501321,"journal":{"name":"arXiv - QuanBio - Cell Behavior","volume":"15 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-07-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141770806","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
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