Lab on a Chip最新文献

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Intelligent optoelectrowetting digital microfluidic system for real-time selective parallel manipulation of biological droplet arrays.
IF 6.1 2区 工程技术
Lab on a Chip Pub Date : 2024-12-11 DOI: 10.1039/d4lc00804a
Tianyi Wang, Shizheng Zhou, Xuekai Liu, Jianghao Zeng, Xiaohan He, Zhihang Yu, Zhiyuan Liu, Xiaomei Liu, Jing Jin, Yonggang Zhu, Liuyong Shi, Hong Yan, Teng Zhou
{"title":"Intelligent optoelectrowetting digital microfluidic system for real-time selective parallel manipulation of biological droplet arrays.","authors":"Tianyi Wang, Shizheng Zhou, Xuekai Liu, Jianghao Zeng, Xiaohan He, Zhihang Yu, Zhiyuan Liu, Xiaomei Liu, Jing Jin, Yonggang Zhu, Liuyong Shi, Hong Yan, Teng Zhou","doi":"10.1039/d4lc00804a","DOIUrl":"https://doi.org/10.1039/d4lc00804a","url":null,"abstract":"<p><p>Optoelectrowetting technology generates virtual electrodes to manipulate droplets by projecting optical patterns onto the photoconductive layer. This method avoids the complex design of the physical circuitry of dielectricwetting chips, compensating for the inability to reconstruct the electrode. However, the current technology relies on operators to manually position the droplets, draw optical patterns, and preset the droplet movement paths. It lacks real-time feedback on droplet information and the ability for independent droplet control, which can lead to droplet miscontrol and contamination. This paper presents a combination of optoelectrowetting with deep learning algorithms, integrating software and a photoelectric detection platform, and develops an optoelectrowetting intelligent control system. First, a target detection algorithm identifies droplet characteristics in real-time and automatically generate virtual electrodes to control movement. Simultaneously, a tracking algorithm outputs trajectories and ID information for efficient droplet arrays tracking. The results show that the system can automatically control the movement and fusion of multiple droplets in parallel and realize the automatic arrangement and storage of disordered droplet arrays without any additional electrodes and sensing devices. Additionally, through the automated control of the system, the cell suspension can be precisely cultured in the specified medium according to experimental requirements, and the growth trend is consistent with that observed in the well plate, significantly enhancing the experiment's flexibility and accuracy. In this paper, we propose an intelligent method applicable to the automated manipulation of discrete droplets. This method would play a crucial role in advancing the applications of digital microfluidic technology in biomedicine and other fields.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" ","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-12-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142805626","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
A 3D-printed multi-compartment organ-on-chip platform with a tubing-free pump models communication with the lymph node.
IF 6.1 2区 工程技术
Lab on a Chip Pub Date : 2024-12-11 DOI: 10.1039/d4lc00489b
Sophie R Cook, Alexander G Ball, Anwaruddin Mohammad, Rebecca R Pompano
{"title":"A 3D-printed multi-compartment organ-on-chip platform with a tubing-free pump models communication with the lymph node.","authors":"Sophie R Cook, Alexander G Ball, Anwaruddin Mohammad, Rebecca R Pompano","doi":"10.1039/d4lc00489b","DOIUrl":"https://doi.org/10.1039/d4lc00489b","url":null,"abstract":"<p><p>Multi-organ-on-chip systems (MOOCs) have the potential to mimic communication between organ systems and reveal mechanisms of health and disease. However, many existing MOOCs are challenging for non-experts to implement due to complex tubing, electronics, or pump mechanisms. In addition, few MOOCs have incorporated immune organs such as the lymph node (LN), limiting their applicability to model critical events such as vaccination. Here we developed a 3D-printed, user-friendly device and companion tubing-free impeller pump with the capacity to co-culture two or more tissue samples, including a LN, under a recirculating common media. Native tissue structure and immune function were incorporated by maintaining slices of murine LN tissue <i>ex vivo</i> in 3D-printed mesh supports for at least 24 h. In a two-compartment model of a LN and an upstream injection site in mock tissue, vaccination of the multi-compartment chip was similar to <i>in vivo</i> vaccination in terms of locations of antigen accumulation and acute changes in activation markers and gene expression in the LN. We anticipate that in the future, this flexible platform will enable models of multi-organ immune responses throughout the body.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" ","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-12-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142805625","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Retinal organoid chip: engineering a physiomimetic oxygen gradient for optimizing long term culture of human retinal organoids.
IF 6.1 2区 工程技术
Lab on a Chip Pub Date : 2024-12-11 DOI: 10.1039/d4lc00771a
Emma Drabbe, Daniel Pelaez, Ashutosh Agarwal
{"title":"Retinal organoid chip: engineering a physiomimetic oxygen gradient for optimizing long term culture of human retinal organoids.","authors":"Emma Drabbe, Daniel Pelaez, Ashutosh Agarwal","doi":"10.1039/d4lc00771a","DOIUrl":"https://doi.org/10.1039/d4lc00771a","url":null,"abstract":"<p><p>An oxygen gradient across the retina plays a crucial role in its development and function. The inner retina resides in a hypoxic environment (2% O<sub>2</sub>) adjacent to the vitreous cavity. Oxygenation levels rapidly increase towards the outer retina (18% O<sub>2</sub>) at the choroid. In addition to retinal stratification, oxygen levels are critical for the health of retinal ganglion cells (RGCs), which relay visual information from the retina to the brain. Human stem cell derived retinal organoids are being engineered to mimic the structure and function of human retina for applications such as disease modeling, development of therapeutics, and cell replacement therapies. However, rapid degeneration of the retinal ganglion cell layers are a common limitation of human retinal organoid platforms. We report the design of a novel retinal organoid chip (ROC) that maintains a physiologically relevant oxygen gradient and allows the maturation of inner and outer retinal cell phenotypes for human retinal organoids. Our PDMS-free ROC holds 55 individual retinal organoids that were manually seeded, cultured for extended periods (over 150 days), imaged <i>in situ</i>, and retrieved. ROC was designed from first principles of liquid and gas mass transport, and fabricated from biologically- and chemically inert materials using rapid prototyping techniques such as micromachining, laser cutting, 3D printing and bonding. After computational and experimental validation of oxygen gradients, human induced pluripotent stem cell derived retinal organoids were transferred into the ROC, differentiated, cultured and imaged within the chip. ROCs that maintained active perfusion and stable oxygen gradients were successful in inducing higher viability of RGCs within retinal organoids than static controls, or ROC without oxygen gradients. Our physiologically relevant and higher-throughput retinal organoid culture system is well suited for applications in studying developmental perturbations to primate retinogenesis, including those driven by inherited traits, fetal environmental exposure to toxic agents, or acquired by genetic mutations, such as retinoblastoma.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" ","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-12-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142805628","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
OoTrap: enhancing oocyte collection and maturation with a field-deployable fluidic device.
IF 6.1 2区 工程技术
Lab on a Chip Pub Date : 2024-12-11 DOI: 10.1039/d4lc00660g
Roksan Franko, Marcia de Almeida Monteiro Melo Ferraz
{"title":"OoTrap: enhancing oocyte collection and maturation with a field-deployable fluidic device.","authors":"Roksan Franko, Marcia de Almeida Monteiro Melo Ferraz","doi":"10.1039/d4lc00660g","DOIUrl":"https://doi.org/10.1039/d4lc00660g","url":null,"abstract":"<p><p>Assisted reproductive technologies (ART) are pivotal for contemporary reproductive medicine and species conservation. However, the manual handling required in these processes introduces stress that can compromise oocyte and embryo quality. This study introduces OoTrap, a novel fluidic device designed to streamline ART workflows by facilitating the capture and maturation of oocytes in a compact unit. The device also reintroduces mechanical forces similar to those in the <i>in vivo</i> environment, which are often missing in conventional systems. OoTrap operates in both static and perfusion-based modes, offering flexibility and optimal conditions for oocyte maturation. Notably, OoTrap achieved higher <i>in vitro</i> maturation (IVM) rates under perfusion, produced oocytes with fewer chromosomal abnormalities, and maintained spindle morphology integrity. The incorporation of a heating system and a 3D-printed syringe pump enabled IVM outside the incubator, making OoTrap suitable for field applications. The results highlight the potential of OoTrap to enhance ART outcomes by reducing manual handling, providing a controlled microenvironment, and offering a practical solution for field-based ART applications.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" ","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-12-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142805627","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Advancing cellular transfer printing: achieving bioadhesion-free deposition via vibration microstreaming.
IF 6.1 2区 工程技术
Lab on a Chip Pub Date : 2024-12-10 DOI: 10.1039/d4lc00601a
Ziyu Huang, Yinning Zhou, Yu Liu, Yue Quan, Qiu Yin, Yucheng Luo, Yimeng Su, Bingpu Zhou, Wenming Zhang, Benpeng Zhu, Zhichao Ma
{"title":"Advancing cellular transfer printing: achieving bioadhesion-free deposition <i>via</i> vibration microstreaming.","authors":"Ziyu Huang, Yinning Zhou, Yu Liu, Yue Quan, Qiu Yin, Yucheng Luo, Yimeng Su, Bingpu Zhou, Wenming Zhang, Benpeng Zhu, Zhichao Ma","doi":"10.1039/d4lc00601a","DOIUrl":"https://doi.org/10.1039/d4lc00601a","url":null,"abstract":"<p><p>Cell transfer printing plays an essential role in biomedical research and clinical diagnostics. Traditional bioadhesion-based methods often necessitate complex surface modifications and offer limited control over the quantity of transferred cells. There is a critical need for a modification-free, non-labeling, and high-throughput cell transfer printing technique. In this study, an adhesion-free cellular transfer printing method based on vibration-induced microstreaming is introduced. By adjusting the volume of the microcavity, the number of cells transferred per microtiter well can be realized to the level of a single cell. Additionally, it allows for precise control of large-scale cellular spatial distribution, leading to the formation of biomimetic patterns. Moreover, the demonstrated biocompatibility and high throughput of this cell transfer printing method highlight its potential utility. The correspondence of the transferred cell amount to the vibration and frequencies allows the system to exhibit excellent tunability of the transferred cell amount and pattern. This bioadhesion-free cell transfer printing method holds promise for advancing cell manipulation in biomedical research and analysis.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" ","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-12-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142798646","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
A gravity-driven microfluidic metering system for automation of multiplexed bioassays.
IF 6.1 2区 工程技术
Lab on a Chip Pub Date : 2024-12-10 DOI: 10.1039/d4lc00800f
Lu Zhang, Johnson Q Cui, Shuhuai Yao
{"title":"A gravity-driven microfluidic metering system for automation of multiplexed bioassays.","authors":"Lu Zhang, Johnson Q Cui, Shuhuai Yao","doi":"10.1039/d4lc00800f","DOIUrl":"https://doi.org/10.1039/d4lc00800f","url":null,"abstract":"<p><p>Automatic and precise fluid manipulation is essential in microfluidic applications. Microfluidic metering, in particular, plays a critical role in achieving the multiplexity of assays, reaction consistency, quantitative analysis, and the scalability of microfluidic operations. However, existing fluid metering techniques often face limitations, such as high complexity, high cost, reliance on external accessories, and lack of precision, which have restricted their use in multiplexed and quantitative analysis, especially in portable applications. In this study, we present a novel portable gravity-driven metering system designed for automated multiplexed fluid metering, multistep fluid control, and multi-chamber signal readout. Our metering chip utilizes gravitational force to dispense sample liquids, allowing for versatile and precise metering. Guided by a series of numerical simulations, we optimized the design of our metering chip to achieve rapid and accurate liquid metering. Furthermore, thermal control valves were employed to facilitate automated and programmable fluid transfer, eliminating the need for external equipment. To enhance user experience, we developed a smartphone-assisted readout pod for seamless integration with the metering chip. We validated the efficacy of our platform through a proof-of-concept multiplexed analysis of urinary biomarkers, demonstrating high sensitivity, specificity, and absolute quantification capabilities. Our gravity-driven metering system shows significant potential for applications in multiplexed diagnostics, drug screening, and material synthesis, effectively addressing critical needs in fluid manipulation and analysis.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" ","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-12-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142798643","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Correction: Distal renal tubular system-on-a-chip for studying the pathogenesis of influenza A virus-induced kidney injury.
IF 6.1 2区 工程技术
Lab on a Chip Pub Date : 2024-12-10 DOI: 10.1039/d4lc90105c
Yueyue Huangfu, Ji Wang, Jiao Feng, Zhi-Ling Zhang
{"title":"Correction: Distal renal tubular system-on-a-chip for studying the pathogenesis of influenza A virus-induced kidney injury.","authors":"Yueyue Huangfu, Ji Wang, Jiao Feng, Zhi-Ling Zhang","doi":"10.1039/d4lc90105c","DOIUrl":"https://doi.org/10.1039/d4lc90105c","url":null,"abstract":"<p><p>Correction for 'Distal renal tubular system-on-a-chip for studying the pathogenesis of influenza A virus-induced kidney injury' by Yueyue Huangfu <i>et al.</i>, <i>Lab Chip</i>, 2023, 23, 4255-4264, https://doi.org/10.1039/D3LC00616F.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" ","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-12-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142798648","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Rapid automated production of tubular 3D intestine-on-a-chip with diverse cell types using coaxial bioprinting.
IF 6.1 2区 工程技术
Lab on a Chip Pub Date : 2024-12-09 DOI: 10.1039/d4lc00731j
Heeju Song, Yeonjin Hong, Hyungseok Lee
{"title":"Rapid automated production of tubular 3D intestine-on-a-chip with diverse cell types using coaxial bioprinting.","authors":"Heeju Song, Yeonjin Hong, Hyungseok Lee","doi":"10.1039/d4lc00731j","DOIUrl":"https://doi.org/10.1039/d4lc00731j","url":null,"abstract":"<p><p>Despite considerable animal sacrifices and investments, drug development often falters in clinical trials due to species differences. To address this issue, specific <i>in vitro</i> models, such as organ-on-a-chip technology using human cells in microfluidic devices, are recognized as promising alternatives. Among the various organs, the human small intestine plays a pivotal role in drug development, particularly in the assessment of digestion and nutrient absorption. However, current intestine-on-a-chip devices struggle to accurately replicate the complex 3D tubular structures of the human small intestine, particularly when it comes to integrating a variety of cell types effectively. This limitation is primarily due to conventional fabrication methods, such as soft lithography and replica molding. In this research, we introduce a novel coaxial bioprinting method to construct 3D tubular structures that closely emulate the organization and functionality of the small intestine with multiple cell types. To ensure stable production of these small intestine-like tubular structures, we analyzed the rheological properties of bioinks to select the most suitable materials for coaxial bioprinting technology. Additionally, we conducted biological assessments to validate the gene expression patterns and functional attributes of the 3D intestine-on-a-chip. Our 3D intestine-on-a-chip, which faithfully replicates intestinal functions and organization, demonstrates clear superiority in both structure and biological function compared to the conventional 2D model. This innovative approach holds significant promise for a wide range of future applications.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" ","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-12-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142793997","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Flow cell for high throughput Raman spectroscopy of non-transparent solutions.
IF 6.1 2区 工程技术
Lab on a Chip Pub Date : 2024-12-04 DOI: 10.1039/d4lc00586d
Filippo Zorzi, Emil Alstrup Jensen, Murat Serhatlioglu, Silvio Bonfadini, Morten Hanefeld Dziegiel, Luigino Criante, Anders Kristensen
{"title":"Flow cell for high throughput Raman spectroscopy of non-transparent solutions.","authors":"Filippo Zorzi, Emil Alstrup Jensen, Murat Serhatlioglu, Silvio Bonfadini, Morten Hanefeld Dziegiel, Luigino Criante, Anders Kristensen","doi":"10.1039/d4lc00586d","DOIUrl":"https://doi.org/10.1039/d4lc00586d","url":null,"abstract":"<p><p>This work introduces a high-throughput setup for Raman analysis of various flowing fluids, both transparent and non-transparent. The setup employs a microfluidic cell, used with an external optical setup, to control the sample flow's position and dimensions <i>via</i> 3-dimensional hydrodynamic focusing. This approach, in contrast to the prevalent use of fused silica capillaries, reduces the risk of sample photodegradation and boosts measurement efficiency, enhancing overall system throughput. The microfluidic cell has been further evolved to laminate two distinct flows from different samples in parallel. Using line excitation, both samples can be simultaneously excited without moving parts, further increasing throughput. This setup also enables real-time monitoring of phenomena like mixing or potential reactions between the two fluids. This development could significantly advance the creation of highly sensitive, high-throughput sensors for fluid composition analysis.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" ","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142764712","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
An integrated microfluidic device for sorting of tumor organoids using image recognition.
IF 6.1 2区 工程技术
Lab on a Chip Pub Date : 2024-12-04 DOI: 10.1039/d4lc00746h
Xingyang Yan, Deng Tan, Lei Yu, Danyu Li, Zhenghao Wang, Weiren Huang, Hongkai Wu
{"title":"An integrated microfluidic device for sorting of tumor organoids using image recognition.","authors":"Xingyang Yan, Deng Tan, Lei Yu, Danyu Li, Zhenghao Wang, Weiren Huang, Hongkai Wu","doi":"10.1039/d4lc00746h","DOIUrl":"https://doi.org/10.1039/d4lc00746h","url":null,"abstract":"<p><p>Tumor organoids present a challenge in drug screening due to their considerable heterogeneity in morphology and size. To address this issue, we proposed a portable microfluidic device that employs image processing algorithms for specific target organoid recognition and microvalve-controlled deflection for sorting and collection. This morphology-activated organoid sorting system offers numerous advantages, such as automated classification, portability, low cost, label-free sample preparation, and gentle handling of organoids. We conducted classification experiments using polystyrene beads, F9 tumoroids and patient-derived tumor organoids, achieving organoid separation efficiency exceeding 88%, purity surpassing 91%, viability exceeding 97% and classification throughput of 800 per hour, thereby meeting the demands of clinical organoid medicine.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" ","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142764709","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","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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