Annual Review of Biomedical Engineering最新文献

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CRISPR/Cas Technology: The Unique Synthetic Biology Genome-Editing Tool Shifting the Paradigm in Viral Diagnostics, Defense, and Therapeutics. CRISPR/Cas 技术:独特的合成生物学基因组编辑工具,改变了病毒诊断、防御和治疗的范式。
IF 12.8 1区 工程技术
Annual Review of Biomedical Engineering Pub Date : 2024-07-01 Epub Date: 2024-06-20 DOI: 10.1146/annurev-bioeng-081723-013033
Lang Zhou, Aleksandr L Simonian
{"title":"CRISPR/Cas Technology: The Unique Synthetic Biology Genome-Editing Tool Shifting the Paradigm in Viral Diagnostics, Defense, and Therapeutics.","authors":"Lang Zhou, Aleksandr L Simonian","doi":"10.1146/annurev-bioeng-081723-013033","DOIUrl":"10.1146/annurev-bioeng-081723-013033","url":null,"abstract":"<p><p>The emergence of the COVID-19 pandemic has starkly exposed our significantly limited ability to promptly identify and respond to emergent biological threats. Consequently, there is an urgent need to advance biotechnological methods for addressing both known and unforeseen biological hazards. Recently, the CRISPR/Cas system has revolutionized genetic engineering, enabling precise and efficient synthetic biology applications. Therefore, this review aims to provide a comprehensive introduction to the fundamental principles underlying the CRISPR/Cas system and assess the advantages and limitations of various CRISPR/Cas-based techniques applicable to the detection of, defense against, and treatment of viral infections. These techniques include viral diagnostics, the development of antiviral vaccines, B cell engineering for antibody production, viral activation/interference, and epigenetic modifications. Furthermore, this review delves into the challenges and bioethical considerations associated with use of the CRISPR/Cas system. With the continuous evolution of technology, the CRISPR/Cas system holds considerable promise for addressing both existing and unforeseen biological threats.</p>","PeriodicalId":50757,"journal":{"name":"Annual Review of Biomedical Engineering","volume":" ","pages":"247-272"},"PeriodicalIF":12.8,"publicationDate":"2024-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139724899","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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
Histotripsy: A Method for Mechanical Tissue Ablation with Ultrasound. 组织切削术:利用超声波进行机械组织消融的方法。
IF 12.8 1区 工程技术
Annual Review of Biomedical Engineering Pub Date : 2024-07-01 Epub Date: 2024-06-20 DOI: 10.1146/annurev-bioeng-073123-022334
Zhen Xu, Tatiana D Khokhlova, Clifford S Cho, Vera A Khokhlova
{"title":"Histotripsy: A Method for Mechanical Tissue Ablation with Ultrasound.","authors":"Zhen Xu, Tatiana D Khokhlova, Clifford S Cho, Vera A Khokhlova","doi":"10.1146/annurev-bioeng-073123-022334","DOIUrl":"10.1146/annurev-bioeng-073123-022334","url":null,"abstract":"<p><p>Histotripsy is a relatively new therapeutic ultrasound technology to mechanically liquefy tissue into subcellular debris using high-amplitude focused ultrasound pulses. In contrast to conventional high-intensity focused ultrasound thermal therapy, histotripsy has specific clinical advantages: the capacity for real-time monitoring using ultrasound imaging, diminished heat sink effects resulting in lesions with sharp margins, effective removal of the treated tissue, a tissue-selective feature to preserve crucial structures, and immunostimulation. The technology is being evaluated in small and large animal models for treating cancer, thrombosis, hematomas, abscesses, and biofilms; enhancing tumor-specific immune response; and neurological applications. Histotripsy has been recently approved by the US Food and Drug Administration to treat liver tumors, with clinical trials undertaken for benign prostatic hyperplasia and renal tumors. This review outlines the physical principles of various types of histotripsy; presents major parameters of the technology and corresponding hardware and software, imaging methods, and bioeffects; and discusses the most promising preclinical and clinical applications.</p>","PeriodicalId":50757,"journal":{"name":"Annual Review of Biomedical Engineering","volume":" ","pages":"141-167"},"PeriodicalIF":12.8,"publicationDate":"2024-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139724900","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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
3D Traction Force Microscopy in Biological Gels: From Single Cells to Multicellular Spheroids. 生物凝胶中的三维牵引力显微镜:从单细胞到多细胞球。
IF 12.8 1区 工程技术
Annual Review of Biomedical Engineering Pub Date : 2024-07-01 Epub Date: 2024-06-20 DOI: 10.1146/annurev-bioeng-103122-031130
Brian C H Cheung, Rana J Abbed, Mingming Wu, Susan E Leggett
{"title":"3D Traction Force Microscopy in Biological Gels: From Single Cells to Multicellular Spheroids.","authors":"Brian C H Cheung, Rana J Abbed, Mingming Wu, Susan E Leggett","doi":"10.1146/annurev-bioeng-103122-031130","DOIUrl":"10.1146/annurev-bioeng-103122-031130","url":null,"abstract":"<p><p>Cell traction force plays a critical role in directing cellular functions, such as proliferation, migration, and differentiation. Current understanding of cell traction force is largely derived from 2D measurements where cells are plated on 2D substrates. However, 2D measurements do not recapitulate a vital aspect of living systems; that is, cells actively remodel their surrounding extracellular matrix (ECM), and the remodeled ECM, in return, can have a profound impact on cell phenotype and traction force generation. This reciprocal adaptivity of living systems is encoded in the material properties of biological gels. In this review, we summarize recent progress in measuring cell traction force for cells embedded within 3D biological gels, with an emphasis on cell-ECM cross talk. We also provide perspectives on tools and techniques that could be adapted to measure cell traction force in complex biochemical and biophysical environments.</p>","PeriodicalId":50757,"journal":{"name":"Annual Review of Biomedical Engineering","volume":" ","pages":"93-118"},"PeriodicalIF":12.8,"publicationDate":"2024-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139693513","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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
Direct Electron Transfer-Type Oxidoreductases for Biomedical Applications. 生物医学应用中的直接电子转移型氧化还原酶。
IF 12.8 1区 工程技术
Annual Review of Biomedical Engineering Pub Date : 2024-07-01 Epub Date: 2024-06-20 DOI: 10.1146/annurev-bioeng-110222-101926
Keisei Sowa, Junko Okuda-Shimazaki, Eole Fukawa, Koji Sode
{"title":"Direct Electron Transfer-Type Oxidoreductases for Biomedical Applications.","authors":"Keisei Sowa, Junko Okuda-Shimazaki, Eole Fukawa, Koji Sode","doi":"10.1146/annurev-bioeng-110222-101926","DOIUrl":"10.1146/annurev-bioeng-110222-101926","url":null,"abstract":"<p><p>Among the various types of enzyme-based biosensors, sensors utilizing enzymes capable of direct electron transfer (DET) are recognized as the most ideal. However, only a limited number of redox enzymes are capable of DET with electrodes, that is, dehydrogenases harboring a subunit or domain that functions specifically to accept electrons from the redox cofactor of the catalytic site and transfer the electrons to the external electron acceptor. Such subunits or domains act as built-in mediators for electron transfer between enzymes and electrodes; consequently, such enzymes enable direct electron transfer to electrodes and are designated as DET-type enzymes. DET-type enzymes fall into several categories, including redox cofactors of catalytic reactions, built-in mediators for DET with electrodes and by their protein hierarchic structures, DET-type oxidoreductases with oligomeric structures harboring electron transfer subunits, and monomeric DET-type oxidoreductases harboring electron transfer domains. In this review, we cover the science of DET-type oxidoreductases and their biomedical applications. First, we introduce the structural biology and current understanding of DET-type enzyme reactions. Next, we describe recent technological developments based on DET-type enzymes for biomedical applications, such as biosensors and biochemical energy harvesting for self-powered medical devices. Finally, after discussing how to further engineer and create DET-type enzymes, we address the future prospects for DET-type enzymes in biomedical engineering.</p>","PeriodicalId":50757,"journal":{"name":"Annual Review of Biomedical Engineering","volume":" ","pages":"357-382"},"PeriodicalIF":12.8,"publicationDate":"2024-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139998219","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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
Kidney Disease Modeling with Organoids and Organs-on-Chips. 利用有机体和芯片器官建立肾脏疾病模型
IF 12.8 1区 工程技术
Annual Review of Biomedical Engineering Pub Date : 2024-07-01 Epub Date: 2024-06-20 DOI: 10.1146/annurev-bioeng-072623-044010
Samira Musah, Rohan Bhattacharya, Jonathan Himmelfarb
{"title":"Kidney Disease Modeling with Organoids and Organs-on-Chips.","authors":"Samira Musah, Rohan Bhattacharya, Jonathan Himmelfarb","doi":"10.1146/annurev-bioeng-072623-044010","DOIUrl":"10.1146/annurev-bioeng-072623-044010","url":null,"abstract":"<p><p>Kidney disease is a global health crisis affecting more than 850 million people worldwide. In the United States, annual Medicare expenditures for kidney disease and organ failure exceed $81 billion. Efforts to develop targeted therapeutics are limited by a poor understanding of the molecular mechanisms underlying human kidney disease onset and progression. Additionally, 90% of drug candidates fail in human clinical trials, often due to toxicity and efficacy not accurately predicted in animal models. The advent of ex vivo kidney models, such as those engineered from induced pluripotent stem (iPS) cells and organ-on-a-chip (organ-chip) systems, has garnered considerable interest owing to their ability to more accurately model tissue development and patient-specific responses and drug toxicity. This review describes recent advances in developing kidney organoids and organ-chips by harnessing iPS cell biology to model human-specific kidney functions and disease states. We also discuss challenges that must be overcome to realize the potential of organoids and organ-chips as dynamic and functional conduits of the human kidney. Achieving these technological advances could revolutionize personalized medicine applications and therapeutic discovery for kidney disease.</p>","PeriodicalId":50757,"journal":{"name":"Annual Review of Biomedical Engineering","volume":" ","pages":"383-414"},"PeriodicalIF":12.8,"publicationDate":"2024-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11479997/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139998220","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Recent Developments in Aerosol Pulmonary Drug Delivery: New Technologies, New Cargos, and New Targets. 肺部气溶胶给药的最新发展:新技术、新载体和新目标。
IF 12.8 1区 工程技术
Annual Review of Biomedical Engineering Pub Date : 2024-07-01 Epub Date: 2024-06-20 DOI: 10.1146/annurev-bioeng-110122-010848
Ian R Woodward, Catherine A Fromen
{"title":"Recent Developments in Aerosol Pulmonary Drug Delivery: New Technologies, New Cargos, and New Targets.","authors":"Ian R Woodward, Catherine A Fromen","doi":"10.1146/annurev-bioeng-110122-010848","DOIUrl":"10.1146/annurev-bioeng-110122-010848","url":null,"abstract":"<p><p>There is nothing like a global pandemic to motivate the need for improved respiratory treatments and mucosal vaccines. Stimulated by the COVID-19 pandemic, pulmonary aerosol drug delivery has seen a flourish of activity, building on the prior decades of innovation in particle engineering, inhaler device technologies, and clinical understanding. As such, the field has expanded into new directions and is working toward the efficient delivery of increasingly complex cargos to address a wider range of respiratory diseases. This review seeks to highlight recent innovations in approaches to personalize inhalation drug delivery, deliver complex cargos, and diversify the targets treated and prevented through pulmonary drug delivery. We aim to inform readers of the emerging efforts within the field and predict where future breakthroughs are expected to impact the treatment of respiratory diseases.</p>","PeriodicalId":50757,"journal":{"name":"Annual Review of Biomedical Engineering","volume":" ","pages":"307-330"},"PeriodicalIF":12.8,"publicationDate":"2024-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11222059/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139998221","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Electronic Skin: Opportunities and Challenges in Convergence with Machine Learning. 电子皮肤:与机器学习融合的机遇与挑战。
IF 12.8 1区 工程技术
Annual Review of Biomedical Engineering Pub Date : 2024-07-01 DOI: 10.1146/annurev-bioeng-103122-032652
Ja Hoon Koo, Young Joong Lee, Hye Jin Kim, Wojciech Matusik, Dae-Hyeong Kim, Hyoyoung Jeong
{"title":"Electronic Skin: Opportunities and Challenges in Convergence with Machine Learning.","authors":"Ja Hoon Koo, Young Joong Lee, Hye Jin Kim, Wojciech Matusik, Dae-Hyeong Kim, Hyoyoung Jeong","doi":"10.1146/annurev-bioeng-103122-032652","DOIUrl":"https://doi.org/10.1146/annurev-bioeng-103122-032652","url":null,"abstract":"<p><p>Recent advancements in soft electronic skin (e-skin) have led to the development of human-like devices that reproduce the skin's functions and physical attributes. These devices are being explored for applications in robotic prostheses as well as for collecting biopotentials for disease diagnosis and treatment, as exemplified by biomedical e-skins. More recently, machine learning (ML) has been utilized to enhance device control accuracy and data processing efficiency. The convergence of e-skin technologies with ML is promoting their translation into clinical practice, especially in healthcare. This review highlights the latest developments in ML-reinforced e-skin devices for robotic prostheses and biomedical instrumentations. We first describe technological breakthroughs in state-of-the-art e-skin devices, emphasizing technologies that achieve skin-like properties. We then introduce ML methods adopted for control optimization and pattern recognition, followed by practical applications that converge the two technologies. Lastly, we briefly discuss the challenges this interdisciplinary research encounters in its clinical and industrial transition.</p>","PeriodicalId":50757,"journal":{"name":"Annual Review of Biomedical Engineering","volume":"26 1","pages":"331-355"},"PeriodicalIF":12.8,"publicationDate":"2024-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141499598","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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
Fly Me to the Micron: Microtechnologies for Drosophila Research. 让我飞向微米:果蝇研究的微技术》。
IF 12.8 1区 工程技术
Annual Review of Biomedical Engineering Pub Date : 2024-07-01 DOI: 10.1146/annurev-bioeng-050423-054647
Utku M Sonmez, Nolan Frey, Philip R LeDuc, Jonathan S Minden
{"title":"Fly Me to the Micron: Microtechnologies for <i>Drosophila</i> Research.","authors":"Utku M Sonmez, Nolan Frey, Philip R LeDuc, Jonathan S Minden","doi":"10.1146/annurev-bioeng-050423-054647","DOIUrl":"https://doi.org/10.1146/annurev-bioeng-050423-054647","url":null,"abstract":"<p><p>Multicellular model organisms, such as <i>Drosophila melanogaster</i> (fruit fly), are frequently used in a myriad of biological research studies due to their biological significance and global standardization. However, traditional tools used in these studies generally require manual handling, subjective phenotyping, and bulk treatment of the organisms, resulting in laborious experimental protocols with limited accuracy. Advancements in microtechnology over the course of the last two decades have allowed researchers to develop automated, high-throughput, and multifunctional experimental tools that enable novel experimental paradigms that would not be possible otherwise. We discuss recent advances in microtechnological systems developed for small model organisms using <i>D. melanogaster</i> as an example. We critically analyze the state of the field by comparing the systems produced for different applications. Additionally, we suggest design guidelines, operational tips, and new research directions based on the technical and knowledge gaps in the literature. This review aims to foster interdisciplinary work by helping engineers to familiarize themselves with model organisms while presenting the most recent advances in microengineering strategies to biologists.</p>","PeriodicalId":50757,"journal":{"name":"Annual Review of Biomedical Engineering","volume":"26 1","pages":"441-473"},"PeriodicalIF":12.8,"publicationDate":"2024-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141499599","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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
Medical Microrobots 医疗微型机器人
IF 9.7 1区 工程技术
Annual Review of Biomedical Engineering Pub Date : 2024-04-10 DOI: 10.1146/annurev-bioeng-081523-033131
Veronica Iacovacci, Eric Diller, Daniel Ahmed, Arianna Menciassi
{"title":"Medical Microrobots","authors":"Veronica Iacovacci, Eric Diller, Daniel Ahmed, Arianna Menciassi","doi":"10.1146/annurev-bioeng-081523-033131","DOIUrl":"https://doi.org/10.1146/annurev-bioeng-081523-033131","url":null,"abstract":"Scientists around the world have long aimed to produce miniature robots that can be controlled inside the human body to aid doctors in identifying and treating diseases. Such microrobots hold the potential to access hard-to-reach areas of the body through the natural lumina. Wireless access has the potential to overcome drawbacks of systemic therapy, as well as to enable completely new minimally invasive procedures. The aim of this review is fourfold: first, to provide a collection of valuable anatomical and physiological information on the target working environments together with engineering tools for the design of medical microrobots; second, to provide a comprehensive updated survey of the technological state of the art in relevant classes of medical microrobots; third, to analyze currently available tracking and closed loop control strategies compatible with the in-body environment; and fourth, to explore the challenges still in place, to steer and inspire future research.","PeriodicalId":50757,"journal":{"name":"Annual Review of Biomedical Engineering","volume":"15 1","pages":""},"PeriodicalIF":9.7,"publicationDate":"2024-04-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140584920","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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
Patient-Specific, Mechanistic Models of Tumor Growth Incorporating Artificial Intelligence and Big Data 结合人工智能和大数据的肿瘤生长患者特异性机制模型
IF 9.7 1区 工程技术
Annual Review of Biomedical Engineering Pub Date : 2024-04-10 DOI: 10.1146/annurev-bioeng-081623-025834
Guillermo Lorenzo, Syed Rakin Ahmed, David A. Hormuth II, Brenna Vaughn, Jayashree Kalpathy-Cramer, Luis Solorio, Thomas E. Yankeelov, Hector Gomez
{"title":"Patient-Specific, Mechanistic Models of Tumor Growth Incorporating Artificial Intelligence and Big Data","authors":"Guillermo Lorenzo, Syed Rakin Ahmed, David A. Hormuth II, Brenna Vaughn, Jayashree Kalpathy-Cramer, Luis Solorio, Thomas E. Yankeelov, Hector Gomez","doi":"10.1146/annurev-bioeng-081623-025834","DOIUrl":"https://doi.org/10.1146/annurev-bioeng-081623-025834","url":null,"abstract":"Despite the remarkable advances in cancer diagnosis, treatment, and management over the past decade, malignant tumors remain a major public health problem. Further progress in combating cancer may be enabled by personalizing the delivery of therapies according to the predicted response for each individual patient. The design of personalized therapies requires the integration of patient-specific information with an appropriate mathematical model of tumor response. A fundamental barrier to realizing this paradigm is the current lack of a rigorous yet practical mathematical theory of tumor initiation, development, invasion, and response to therapy. We begin this review with an overview of different approaches to modeling tumor growth and treatment, including mechanistic as well as data-driven models based on big data and artificial intelligence. We then present illustrative examples of mathematical models manifesting their utility and discuss the limitations of stand-alone mechanistic and data-driven models. We then discuss the potential of mechanistic models for not only predicting but also optimizing response to therapy on a patient-specific basis. We describe current efforts and future possibilities to integrate mechanistic and data-driven models. We conclude by proposing five fundamental challenges that must be addressed to fully realize personalized care for cancer patients driven by computational models.","PeriodicalId":50757,"journal":{"name":"Annual Review of Biomedical Engineering","volume":"94 1","pages":""},"PeriodicalIF":9.7,"publicationDate":"2024-04-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140585094","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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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