ACS Synthetic Biology最新文献_第8页

Optimization of Exon-Skipping Riboswitches and Their Applications to Control Mammalian Cell Fate 优化外显子跳转核糖开关及其在控制哺乳动物细胞命运中的应用

IF 3.7 2区生物学

ACS Synthetic Biology Pub Date : 2024-09-24 DOI: 10.1021/acssynbio.4c0029510.1021/acssynbio.4c00295

Yoko Nomura, Narae Kim, Bochen Zhu, Muhammad Hamzah, Haifeng Zhang and Yohei Yokobayashi*,

{"title":"Optimization of Exon-Skipping Riboswitches and Their Applications to Control Mammalian Cell Fate","authors":"Yoko Nomura, Narae Kim, Bochen Zhu, Muhammad Hamzah, Haifeng Zhang and Yohei Yokobayashi*, ","doi":"10.1021/acssynbio.4c0029510.1021/acssynbio.4c00295","DOIUrl":"https://doi.org/10.1021/acssynbio.4c00295https://doi.org/10.1021/acssynbio.4c00295","url":null,"abstract":"Mammalian riboswitches that can regulate transgene expression via RNA–small molecule interaction have promising applications in medicine and biotechnology, as they involve no protein factors that can induce immunogenic reactions and are not dependent on specially engineered promoters. However, the lack of cell-permeable and low-toxicity small molecules and cognate aptamers that can be exploited as riboswitches and the modest switching performance of mammalian riboswitches have limited their applications. In this study, we systematically optimized the design of a riboswitch that regulates exon skipping via an RNA aptamer that binds ASP2905. We examined two design strategies to modulate the stability of the aptamer base stem that blocks the 5′ splice site to fine-tune the riboswitch characteristics. Furthermore, an optimized riboswitch was used to generate a mouse embryonic stem cell line that can be chemically induced to differentiate into myogenic cells by activating Myod1 expression and a human embryonic kidney cell line that can be induced to trigger apoptosis by activating BAX expression. The results demonstrate the tight chemical regulation of transgenes in mammalian cells to control their phenotype without exogenous protein factors.","PeriodicalId":26,"journal":{"name":"ACS Synthetic Biology","volume":null,"pages":null},"PeriodicalIF":3.7,"publicationDate":"2024-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/acssynbio.4c00295","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142450541","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Impact of Chromatin Organization and Epigenetics on CRISPR-Cas and TALEN Genome Editing 染色质组织和表观遗传学对 CRISPR-Cas 和 TALEN 基因组编辑的影响

IF 3.7 2区生物学

ACS Synthetic Biology Pub Date : 2024-09-24 DOI: 10.1021/acssynbio.4c0009910.1021/acssynbio.4c00099

Surbhi Jain, Guanhua Xun and Huimin Zhao*,

{"title":"Impact of Chromatin Organization and Epigenetics on CRISPR-Cas and TALEN Genome Editing","authors":"Surbhi Jain, Guanhua Xun and Huimin Zhao*, ","doi":"10.1021/acssynbio.4c0009910.1021/acssynbio.4c00099","DOIUrl":"https://doi.org/10.1021/acssynbio.4c00099https://doi.org/10.1021/acssynbio.4c00099","url":null,"abstract":"DNA lies at the heart of the central dogma of life. Altering DNA can modify the flow of information in fundamental cellular processes such as transcription and translation. The ability to precisely manipulate DNA has led to remarkable advances in treating incurable human genetic ailments and has changed the landscape of biological research. Genome editors such as CRISPR-Cas nucleases and TALENs have become ubiquitous tools in basic and applied biological research and have been translated to the clinic to treat patients. The specificity and modularity of these genome editors have made it possible to efficiently engineer genomic DNA; however, underlying principles governing editing outcomes in eukaryotes are still being uncovered. Editing efficiency can vary from cell type to cell type for the same DNA target sequence, necessitating de novo design and validation efforts. Chromatin structure and epigenetic modifications have been shown to affect the activity of genome editors because of the role they play in hierarchical organization of the underlying DNA. Understanding the nuclear search mechanism of genome editors and their molecular interactions with higher order chromatin will lead to improved models for predicting precise genome editing outcomes. Insights from such studies will unlock the entire genome to be engineered for the creation of novel therapies to treat critical illnesses.","PeriodicalId":26,"journal":{"name":"ACS Synthetic Biology","volume":null,"pages":null},"PeriodicalIF":3.7,"publicationDate":"2024-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142449024","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Construction of Whole Cell Bacterial Biosensors as an Alternative Environmental Monitoring Technology to Detect Naphthenic Acids in Oil Sands Process-Affected Water 构建全细胞细菌生物传感器，作为检测油砂加工影响水中环烷酸的替代环境监测技术

IF 3.7 2区生物学

ACS Synthetic Biology Pub Date : 2024-09-23 DOI: 10.1021/acssynbio.4c0026010.1021/acssynbio.4c00260

Tyson Bookout, Steve Shideler, Evan Cooper, Kira Goff, John V. Headley, Lisa M. Gieg and Shawn Lewenza*,

{"title":"Construction of Whole Cell Bacterial Biosensors as an Alternative Environmental Monitoring Technology to Detect Naphthenic Acids in Oil Sands Process-Affected Water","authors":"Tyson Bookout, Steve Shideler, Evan Cooper, Kira Goff, John V. Headley, Lisa M. Gieg and Shawn Lewenza*, ","doi":"10.1021/acssynbio.4c0026010.1021/acssynbio.4c00260","DOIUrl":"https://doi.org/10.1021/acssynbio.4c00260https://doi.org/10.1021/acssynbio.4c00260","url":null,"abstract":"After extraction of bitumen from oil sands deposits, the oil sand process-affected water (OSPW) is stored in tailings ponds. Naphthenic acids (NA) in tailings ponds have been identified as the primary contributor to toxicity to aquatic life. As an alternative to other analytical methods, here we identify bacterial genes induced after growth in naphthenic acids and use synthetic biology approaches to construct a panel of candidate biosensors for NA detection in water. The main promoters of interest were the atuAR promoters from a naphthenic acid degradation operon and upstream TetR regulator, the marR operon which includes a MarR regulator and downstream naphthenic acid resistance genes, and a hypothetical gene with a possible role in fatty acid biology. Promoters were printed and cloned as transcriptional lux reporter plasmids that were introduced into a tailings pond-derived Pseudomonas species. All candidate biosensor strains were tested for transcriptional responses to naphthenic acid mixtures and individual compounds. The three priority promoters respond in a dose-dependent manner to simple, acyclic, and complex NA mixtures, and each promoter has unique NA specificities. The limits of NA detection from the various NA mixtures ranged between 1.5 and 15 mg/L. The atuA and marR promoters also detected NA in small volumes of OSPW samples and were induced by extracts of the panel of OSPW samples. While biosensors have been constructed for other hydrocarbons, here we describe a biosensor approach that could be employed in environmental monitoring of naphthenic acids in oil sands mining wastewater.","PeriodicalId":26,"journal":{"name":"ACS Synthetic Biology","volume":null,"pages":null},"PeriodicalIF":3.7,"publicationDate":"2024-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142450438","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Implementing Optogenetic-Controlled Bacterial Systems in Drosophila melanogaster for Alleviation of Heavy Metal Poisoning 在黑腹果蝇体内实施光遗传学控制细菌系统以减轻重金属中毒症状

IF 3.7 2区生物学

ACS Synthetic Biology Pub Date : 2024-09-23 DOI: 10.1021/acssynbio.4c0040910.1021/acssynbio.4c00409

Junyi Wang, Ying Li, Dawei Sun, Jingyi Li, Lianyue Li, Xinyu Zhang, Xinyu Liu, Zhijie Feng, Huimin Xue, Yuhui Cui, Yiwen Wang*, Duo Liu* and Hanjie Wang*,

{"title":"Implementing Optogenetic-Controlled Bacterial Systems in Drosophila melanogaster for Alleviation of Heavy Metal Poisoning","authors":"Junyi Wang, Ying Li, Dawei Sun, Jingyi Li, Lianyue Li, Xinyu Zhang, Xinyu Liu, Zhijie Feng, Huimin Xue, Yuhui Cui, Yiwen Wang*, Duo Liu* and Hanjie Wang*, ","doi":"10.1021/acssynbio.4c0040910.1021/acssynbio.4c00409","DOIUrl":"https://doi.org/10.1021/acssynbio.4c00409https://doi.org/10.1021/acssynbio.4c00409","url":null,"abstract":"Drosophila melanogaster (fruit fly) is an animal model chassis in biological and genetic research owing to its short life cycle, ease of cultivation, and acceptability to genetic modification. While the D. melanogaster chassis offers valuable insights into drug efficacy, toxicity, and mechanisms, several obvious challenges such as dosage control and drug resistance still limit its utility in pharmacological studies. Our research combines optogenetic control with engineered gut bacteria to facilitate the precise delivery of therapeutic substances in D. melanogaster for biomedical research. We have shown that the engineered bacteria can be orally administered to D. melanogaster to get a stable density of approximately 28,000 CFUs/per fly, leading to no detectable negative effects on the growth of D. melanogaster. In a model of D. melanogaster exposure to heavy metal, these orally administered bacteria uniformly express target genes under green light control to produce MtnB protein for binding and detoxifying lead, which significantly reduces the level of oxidative stress in the intestinal tract of Pb-treated flies. This pioneering study lays the groundwork for using optogenetic-controlled bacteria in the model chassis D. melanogaster to advance biomedical applications.","PeriodicalId":26,"journal":{"name":"ACS Synthetic Biology","volume":null,"pages":null},"PeriodicalIF":3.7,"publicationDate":"2024-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142450436","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Design and Characterization of a Transcriptional Repression Toolkit for Plants 植物转录抑制工具包的设计与表征

IF 3.7 2区生物学

ACS Synthetic Biology Pub Date : 2024-09-23 DOI: 10.1021/acssynbio.4c0040410.1021/acssynbio.4c00404

Kasey Markel, Jean Sabety, Shehan Wijesinghe and Patrick M. Shih*,

{"title":"Design and Characterization of a Transcriptional Repression Toolkit for Plants","authors":"Kasey Markel, Jean Sabety, Shehan Wijesinghe and Patrick M. Shih*, ","doi":"10.1021/acssynbio.4c0040410.1021/acssynbio.4c00404","DOIUrl":"https://doi.org/10.1021/acssynbio.4c00404https://doi.org/10.1021/acssynbio.4c00404","url":null,"abstract":"Regulation of gene expression is essential for all life. Tools to manipulate the gene expression level have therefore proven to be very valuable in efforts to engineer biological systems. However, there are few well-characterized genetic parts that reduce gene expression in plants, commonly known as transcriptional repressors. We characterized the repression activity of a library consisting of repression motifs from approximately 25% of the members of the largest known family of repressors. Combining sequence information with our trans-regulatory function data, we next generated a library of synthetic transcriptional repression motifs with function predicted in advance. After characterizing our synthetic library, we demonstrated not only that many of our synthetic constructs were functional as repressors but also that our advance predictions of repression strength were better than random guesses. Finally, we assessed the functionality of known transcriptional repression motifs from a wide range of eukaryotes. Our study represents the largest plant repressor motif library experimentally characterized to date, providing unique opportunities for tuning transcription in plants.","PeriodicalId":26,"journal":{"name":"ACS Synthetic Biology","volume":null,"pages":null},"PeriodicalIF":3.7,"publicationDate":"2024-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/acssynbio.4c00404","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142450435","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Modular Cloning Tools for Streptomyces spp. and Application to the De Novo Biosynthesis of Flavokermesic Acid 链霉菌的模块化克隆工具及其在黄酮苷酸新生物合成中的应用

IF 3.7 2区生物学

ACS Synthetic Biology Pub Date : 2024-09-22 DOI: 10.1021/acssynbio.4c0041210.1021/acssynbio.4c00412

Jean-Malo Massicard, Delphine Noel, Andrea Calderari, André Le Jeune, Cyrille Pauthenier and Kira J. Weissman*,

{"title":"Modular Cloning Tools for Streptomyces spp. and Application to the De Novo Biosynthesis of Flavokermesic Acid","authors":"Jean-Malo Massicard, Delphine Noel, Andrea Calderari, André Le Jeune, Cyrille Pauthenier and Kira J. Weissman*, ","doi":"10.1021/acssynbio.4c0041210.1021/acssynbio.4c00412","DOIUrl":"https://doi.org/10.1021/acssynbio.4c00412https://doi.org/10.1021/acssynbio.4c00412","url":null,"abstract":"The filamentous Streptomyces are among the most prolific producers of bioactive natural products and are thus attractive chassis for the heterologous expression of native and designed biosynthetic pathways. Although suitable Streptomyces hosts exist, including genetically engineered cluster-free mutants, the approach is currently limited by the relative paucity of synthetic biology tools facilitating the de novo assembly of multicomponent gene clusters. Here, we report a modular system (MoClo) for Streptomyces including a set of adapted vectors and genetic elements, which allow for the construction of complete genetic circuits. Critical functional validation of each of the elements was obtained using the previously reported β-glucuronidase (GusA) reporter system. Furthermore, we provide proof-of-principle for the toolbox inS. albus, demonstrating the efficient assembly of a biosynthetic pathway to flavokermesic acid (FK), an advanced precursor of the commercially valuable carminic acid.","PeriodicalId":26,"journal":{"name":"ACS Synthetic Biology","volume":null,"pages":null},"PeriodicalIF":3.7,"publicationDate":"2024-09-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142450432","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Fundamentals and Exceptions of the LysR-type Transcriptional Regulators LysR 型转录调节因子的基本原理和例外情况

IF 3.7 2区生物学

ACS Synthetic Biology Pub Date : 2024-09-22 DOI: 10.1021/acssynbio.4c0021910.1021/acssynbio.4c00219

Wouter Demeester, Brecht De Paepe and Marjan De Mey*,

{"title":"Fundamentals and Exceptions of the LysR-type Transcriptional Regulators","authors":"Wouter Demeester, Brecht De Paepe and Marjan De Mey*, ","doi":"10.1021/acssynbio.4c0021910.1021/acssynbio.4c00219","DOIUrl":"https://doi.org/10.1021/acssynbio.4c00219https://doi.org/10.1021/acssynbio.4c00219","url":null,"abstract":"LysR-type transcriptional regulators (LTTRs) are emerging as a promising group of macromolecules for the field of biosensors. As the largest family of bacterial transcription factors, the LTTRs represent a vast and mostly untapped repertoire of sensor proteins. To fully harness these regulators for transcription factor-based biosensor development, it is crucial to understand their underlying mechanisms and functionalities. In the first part, this Review discusses the established model and features of LTTRs. As dual-function regulators, these inducible transcription factors exude precise control over their regulatory targets. In the second part of this Review, an overview is given of the exceptions to the “classic” LTTR model. While a general regulatory mechanism has helped elucidate the intricate regulation performed by LTTRs, it is essential to recognize the variations within the family. By combining this knowledge, characterization of new regulators can be done more efficiently and accurately, accelerating the expansion of transcriptional sensors for biosensor development. Unlocking the pool of LTTRs would significantly expand the currently limited range of detectable molecules and regulatory functions available for the implementation of novel synthetic genetic circuitry.","PeriodicalId":26,"journal":{"name":"ACS Synthetic Biology","volume":null,"pages":null},"PeriodicalIF":3.7,"publicationDate":"2024-09-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142450431","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Approaching Optimal pH Enzyme Prediction with Large Language Models. 利用大型语言模型接近最佳 pH 值酶预测。

IF 3.7 2区生物学

ACS Synthetic Biology Pub Date : 2024-09-20 Epub Date: 2024-08-28 DOI: 10.1021/acssynbio.4c00465

Mark Zaretckii, Pavel Buslaev, Igor Kozlovskii, Alexander Morozov, Petr Popov

{"title":"Approaching Optimal pH Enzyme Prediction with Large Language Models.","authors":"Mark Zaretckii, Pavel Buslaev, Igor Kozlovskii, Alexander Morozov, Petr Popov","doi":"10.1021/acssynbio.4c00465","DOIUrl":"10.1021/acssynbio.4c00465","url":null,"abstract":"Enzymes are widely used in biotechnology due to their ability to catalyze chemical reactions: food making, laundry, pharmaceutics, textile, brewing─all these areas benefit from utilizing various enzymes. Proton concentration (pH) is one of the key factors that define the enzyme functioning and efficiency. Usually there is only a narrow range of pH values where the enzyme is active. This is a common problem in biotechnology to design an enzyme with optimal activity in a given pH range. A large part of this task can be completed in silico, by predicting the optimal pH of designed candidates. The success of such computational methods critically depends on the available data. In this study, we developed a language-model-based approach to predict the optimal pH range from the enzyme sequence. We used different splitting strategies based on sequence similarity, protein family annotation, and enzyme classification to validate the robustness of the proposed approach. The derived machine-learning models demonstrated high accuracy across proteins from different protein families and proteins with lower sequence similarities compared with the training set. The proposed method is fast enough for the high-throughput virtual exploration of protein space for the search for sequences with desired optimal pH levels.","PeriodicalId":26,"journal":{"name":"ACS Synthetic Biology","volume":null,"pages":null},"PeriodicalIF":3.7,"publicationDate":"2024-09-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11421216/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142085835","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Algorithms for Autonomous Formation of Multicellular Shapes from Single Cells. 单细胞自主形成多细胞形状的算法。

IF 3.7 2区生物学

ACS Synthetic Biology Pub Date : 2024-09-20 Epub Date: 2024-08-28 DOI: 10.1021/acssynbio.4c00037

Evan Appleton, Noushin Mehdipour, Tristan Daifuku, Demarcus Briers, Iman Haghighi, Michaël Moret, George Chao, Timothy Wannier, Anush Chiappino-Pepe, Jeremy Huang, Calin Belta, George M Church

引用次数: 0

Advances in Flavonoid and Derivative Biosynthesis: Systematic Strategies for the Construction of Yeast Cell Factories. 黄酮类化合物和衍生物生物合成的进展：构建酵母细胞工厂的系统策略》。

IF 3.7 2区生物学

ACS Synthetic Biology Pub Date : 2024-09-20 Epub Date: 2024-08-15 DOI: 10.1021/acssynbio.4c00383

Jian Wang, Cheng Chen, Qi Guo, Yang Gu, Tian-Qiong Shi

{"title":"Advances in Flavonoid and Derivative Biosynthesis: Systematic Strategies for the Construction of Yeast Cell Factories.","authors":"Jian Wang, Cheng Chen, Qi Guo, Yang Gu, Tian-Qiong Shi","doi":"10.1021/acssynbio.4c00383","DOIUrl":"10.1021/acssynbio.4c00383","url":null,"abstract":"Flavonoids, a significant group of natural polyphenolic compounds, possess a broad spectrum of pharmacological effects. Recent advances in the systematic metabolic engineering of yeast cell factories (YCFs) provide new opportunities for enhanced flavonoid production. Herein, we outline the latest research progress on typical flavonoid products in YCFs. Advanced engineering strategies involved in flavonoid biosynthesis are discussed in detail, including enhancing precursor supply, cofactor engineering, optimizing core pathways, eliminating competitive pathways, relieving transport limitations, and dynamic regulation. Additionally, we highlight the existing problems in the biosynthesis of flavonoid glucosides in yeast, such as endogenous degradation of flavonoid glycosides, substrate promiscuity of UDP-glycosyltransferases, and an insufficient supply of UDP-sugars, with summaries on the corresponding solutions. Discussions also cover other typical postmodifications like prenylation and methylation, and the recent biosynthesis of complex flavonoid compounds in yeast. Finally, a series of advanced technologies are envisioned, i.e., semirational enzyme engineering, ML/DL algorithn, and systems biology, with the aspiration of achieving large-scale industrial production of flavonoid compounds in the future.","PeriodicalId":26,"journal":{"name":"ACS Synthetic Biology","volume":null,"pages":null},"PeriodicalIF":3.7,"publicationDate":"2024-09-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141981168","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0