Jove-Journal of Visualized Experiments最新文献

{"title":"In vitro Cannabis Exposures of Lung Epithelial Cells at the Air-Liquid Interface.","authors":"Emily T Wilson, Ilan B Urovitch, Percival Graham, Rui Liu, Cory S Harris, David H Eidelman, Carolyn J Baglole","doi":"10.3791/68102","DOIUrl":"https://doi.org/10.3791/68102","url":null,"abstract":"<p><p>Cannabis is used by an estimated 192 million people around the world. Most people use cannabis through the inhalation of cannabis smoke, which contains combustion by-products that can negatively affect lung health. Knowledge of these risks has led to a growing interest in cannabis vaporizers, which heat the dry cannabis flower without burning. Vaporizing cannabis still releases cannabinoids for inhalation but heats the plant material at a lower temperature. There is currently no standardized in vitro model for assessing the effects of dry cannabis vapor. Therefore, we established a model for the exposure of lung cell cultures at an air-liquid interface (ALI), whereby cells are apically exposed to vaporized cannabis, thereby more accurately simulating lung epithelial cell physiology. This protocol ensures consistent and reproducible delivery of cannabis vapor to the cell surface, providing a reliable platform for investigating the cellular and molecular impacts of vaporized cannabis. This work is the first to standardize an in vitro cannabis vapor delivery method, which can serve as a benchmark for future preclinical cannabis research.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 220","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144585344","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Incorporating Target Protein Structure Flexibility and Dynamics in Computational Drug Discovery Using Ensemble-Based Docking Analysis.","authors":"Ahmad Fadhlurrahman Ahmad Hidayat, Saharuddin Bin Mohamad","doi":"10.3791/67174","DOIUrl":"https://doi.org/10.3791/67174","url":null,"abstract":"<p><p>The drug discovery process is a rigorous, time-consuming, and expensive operation. The computational approach in drug discovery allows researchers to prioritize the most promising compounds for further testing, which would greatly reduce the required resources, leading to an increment of the overall efficiency in the drug discovery pipelines. Structure-based drug discovery is a common approach that requires the structural information of the target protein in a three-dimensional format. However, the current limitation of most computer-aided drug discovery strategies is their inability to introduce the flexibility and dynamics of the target protein structure during the ligand-protein docking simulation. While both induced fit docking and ensemble-based docking aim to address protein flexibility in the docking procedure, the latter can provide a more comprehensive view of dynamic protein behavior by incorporating multiple conformations throughout the simulation. In this report, we demonstrate and discuss the application of a technique called ensemble-based docking analysis that indirectly introduces the target protein structure's flexibility and dynamics in the molecular docking process. The protein and ligand selected for ensemble-based docking studies were lysozyme and Flovokawain B (FB), respectively. FB has been previously reported to have binding activity with lysozyme. A molecular dynamics (MD) simulation was performed on lysozyme in the presence of water, and the total energy, root-mean-square deviation (RMSD), and root-mean-square fluctuation (RMSF) were examined. Conformation clustering was generated based on several clustering cutoff values and was chosen for additional docking analysis with FB. Cluster no 2 gives the lowest binding energy at -29.37 kJ/mol. Molecular docking images were generated to anticipate the presence of binding forces. By incorporating the structural dynamics of the protein, the ensemble-based docking approach can better capture the range of possible binding scenarios, leading to more reliable predictions of binding outcomes.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 220","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144585355","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Rapid Identification of Neisseria gonorrhoeae, Chlamydia trachomatis, and Ureaplasma urealyticum by RNA Amplification Lateral Flow Assay.","authors":"Ruiqi Deng, Zhiyi Tan, Delan Wang, Xiaoqun Luo, Zhigang Wu, Huajin Chen, Huoqiang Chen, Changlin Zhu","doi":"10.3791/68413","DOIUrl":"https://doi.org/10.3791/68413","url":null,"abstract":"<p><p>Neisseria gonorrhoeae (NG), Chlamydia trachomatis (CT), and Ureaplasma urealyticum (UU) are common pathogens associated with sexually transmitted diseases (STDs). NG is the causative agent of gonorrhea, which can result in various disorders of the genitourinary system, including epididymitis, prostatitis, cervicitis, and infertility. CT is a microorganism responsible for non-gonococcal urethritis and pelvic inflammatory disease. UU is associated with non-gonococcal urethritis, cervicitis, prostatitis, and infertility. Current molecular technologies for the analysis of NG, CT, and UU are often complex and time-consuming. Rapid molecular testing for these pathogens may facilitate the early diagnosis of STDs. The RNA amplification lateral flow assay (RGT) is a method that does not require nucleic acid extraction. In this process, samples are lysed to release nucleic acid fragments, which are then amplified through reverse transcription and transcription. The amplified RNA product is recognized and captured by specific probes, forming an RNA-detection probe-gold probe complex that can be immobilized on a nitrocellulose membrane via lateral flow to produce visible bands. The entire procedure takes less than 1 h. The aim of this study was to evaluate the efficacy of RGT for the rapid detection of NG, CT, and UU. A total of 1,416 samples were included. The consistency of NG, CT, and UU when analyzed by RGT compared to the polymerase chain reaction (PCR) fluorescence probing method was 99.03% (307/310), 99.38% (159/160), and 99.02% (303/306), respectively. The consistency of NG, CT, and UU when analyzed by RGT compared to gene sequencing was 99.17% (238/240), 98.95% (188/190), and 98.30% (173/176), respectively. Compared with the pathogen isolation culture method, the detection rates for NG, CT, and UU assays were 100.00% (9/9, 17/17, 8/8). The high sensitivity and specificity, ease of use, and reduced time requirements make this assay ideal for the rapid and accurate detection of NG, CT, and UU.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 220","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144585390","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of Novel Low-DMSO Cryoprotectant for Peripheral Blood Stem Cell Preservation.","authors":"Zhi Guo, Ming-Xin He, Yi-Huizhi Zhang, Qiang Jiang, Qiang Wang","doi":"10.3791/68275","DOIUrl":"https://doi.org/10.3791/68275","url":null,"abstract":"<p><p>Peripheral blood hematopoietic stem cell (PBHSC) cryopreservation is critical for autologous stem cell transplantation (ASCT), but traditional cryoprotective agents (TCPAs) containing 10% dimethyl sulfoxide (DMSO) raise safety concerns due to toxicity risk. This study aimed to validate a novel low-DMSO cryoprotective agent (CPA, 2% DMSO) for PBHSC preservation at -80 °C, eliminating the need for liquid nitrogen storage. PBHSCs from six donors were divided into CPA and TCPA groups. The CPA was mixed with PBHSCs (1:1 vol/vol) and directly stored at -80 °C. TCPA (10% DMSO + 5% human albumin) underwent gradual cooling (1 °C/min) and liquid nitrogen storage. After 1 month, both groups were thawed in a 37 °C water bath. Cell viability, cytoskeletal integrity (microfilaments/microtubules), mitochondrial activity, and colony-forming capacity were compared. After thawing, PBHSC survival was comparable between CPA (91.29%) and TCPA (90.07%). However, CPA outperformed TCPA in cell viability assays (CPA: 89.38% versus TCPA: 79.55%; p < 0.05). Cytoskeletal analysis revealed intact microfilaments and microtubules in CPA-preserved cells, with structural clarity exceeding TCPA. Mitochondrial activity in CPA-treated cells mirrored fresh PBHSCs, exhibiting 8.5% higher activity than TCPA (p < 0.05) and increased mitochondrial complex numbers. Colony-forming assays further confirmed CPA's superiority, with higher colony counts post-induction. CPA enables safe, convenient PBHSC cryopreservation at -80 °C using ultralow DMSO (2%), eliminating liquid nitrogen reliance. Its enhanced cell viability and mitochondrial preservation suggest clinical advantages by reducing infusion toxicity risks. This protocol offers a transformative strategy for ASCT, optimizing safety and operational efficiency.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 220","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144585336","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"In situ Photo-rheology Monitors Viscoelastic Changes in Photo-responsive Polymer Networks.","authors":"Eleanor L Quirk, Michael C Burroughs, Danielle J Mai","doi":"10.3791/68394","DOIUrl":"https://doi.org/10.3791/68394","url":null,"abstract":"<p><p>In situ photo-rheology is a powerful technique for characterizing the evolution of photo-responsive materials during irradiation. This technique enables time-resolved analysis of the mechanical properties of materials undergoing photo-induced molecular changes, such as during additive manufacturing, photo-polymerization, or photo-stimulation. We present a rheological method to monitor dynamic network formation by photo-responsive star polymers. This article describes procedures to configure a rheometer for simultaneous measurement and irradiation, prepare and load photo-responsive polymer samples, identify appropriate measurement parameters, and analyze time-resolved viscoelasticity. Mechanical and kinetic information provides insight into the dynamics of photo-responsive polymer networks. The goal of this protocol is to provide an example of experimental parameter selection, in situ photo-rheological measurements, and mechanical analysis of photo-responsive materials undergoing irradiation. Additionally, this protocol identifies potential modifications to customize the setup for many possible photo-responsive systems, thereby providing guidelines for future users across the materials science community. In situ photo-rheology improves the understanding of photo-responsive material properties, which is important for engineering materials with predictable processing by additive manufacturing, tunable mechanical performance, and stimuli-responsive functions.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 220","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144585343","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The Design and Application of Inhibiting Peptide for Rapid In vitro Downregulation of Post-Translational Modifications Levels at a Specific Site.","authors":"Shu-Ping Zhang, Jia-Qi Liu, Ning Su, Jie Yang, Zhi-Xia Yang, Dong Zhang, Xian-Zhe Gu","doi":"10.3791/68581","DOIUrl":"https://doi.org/10.3791/68581","url":null,"abstract":"<p><p>Proteins are the primary executors of life activities and are regulated at transcriptional, translational, and post-translational modification (PTM) levels. Moreover, PTM represents a more complicated mechanism for regulating protein activities as a protein generally has various types of PTMs and multiple sites for a specific PTM. Plasmid transfection or lentivirus and adenovirus could be used in initial screening of the functionally important PTM site among all identified sites; however, these methods always face challenges such as low-efficiency in cell and tissue entry, time-consuming and high costs, potential immune reaction, etc. To address this, we recently developed and successfully employed a type of recombined peptide called TAT-PIP, TAT-conjugated PTM inhibitory peptide. TAT-PIP consists of a TAT module that facilitates cell and tissue entry and a PIP module that specifically downregulates the PTM at targeted sites through competitive binding. Here, we present a protocol that outlines the design specifications, quality control standards, and functional verification system of TAT-PIP. In the design section, we describe the consistency, the optional length, specificity, and conservation of TAT-PIP. Next, we introduce the quality testing requirements of TAT-PIP, which guarantee its efficacy and safety. In the application of the TAT-PIP part, we introduced the concentration gradient test of TAT-PIP, the incubation process of the tested cells, and the subsequent phenotypic detection. In summary, we describe an effective method for screening PTM sites by selectively knocking down a specific site and observing the resulting phenotype to infer its function. Due to its low synthetic cost and high efficiency, this method overcomes the limitations of existing technologies, such as plasmid transfection.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 220","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144585395","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Whole-mount Retinal Organoid Visualization with Cellular Resolution.","authors":"Marina Cunquero, Helena Isla-Magrané, Maria Marsal, Maddalen Zufiaurre-Seijo, José García-Arumí, Miguel Ángel Zapata, Anna Duarri, Pablo Loza-Alvarez","doi":"10.3791/68384","DOIUrl":"https://doi.org/10.3791/68384","url":null,"abstract":"<p><p>Retinal organoids derived from human induced pluripotent stem cells are intricate 3D structures that mimic the human retina, offering a powerful platform for studying retinal development, disease mechanisms, and potential therapeutic strategies. Moreover, as they are derived from patients, they are becoming increasingly popular as they hold great promise as a tool for personalized medicine. Unlike conventional 2D cell cultures, retinal organoids preserve the native 3D architecture of the retina, allowing for a more realistic representation and enabling more physiologically relevant studies. However, their structural complexity, high cellular density, and diverse composition present significant challenges for characterization. To address these challenges and enhance our understanding of retinal organoid maturation while preserving the 3D context, we combined optical clearing methods with immunolabeling to visualize the entire structure of whole-mount organoids with confocal microscopy. For this, we employed a clearing method compatible with low- and high-numerical-aperture objectives, facilitating full-volume imaging and capturing certain regions of interest with cellular resolution. Using this approach, we identified the morphology and distribution in 3D of the three main neuron paths responsible for the visual information transmission: cone and rod photoreceptors, bipolar and ganglion cells. Our findings shed more light on the visualization techniques to address the spatial organization of retinal cells within the organoid.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 220","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144585396","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Assessment of Physical Activity Intensity with Accelerometers and Oxygen Consumption.","authors":"Matías Briones-Cayupán, Emily Pozo-Santander, Magdalena Vega-Raviola, José Fernández-Fuentes, Paulina Ibacache-Saavedra, Marcelo Cano-Cappellacci","doi":"10.3791/67864","DOIUrl":"https://doi.org/10.3791/67864","url":null,"abstract":"<p><p>The study compares objective methods of assessing the intensity of physical activity. The goal was to identify the differences in physical activity intensity measured through indirect calorimetry and triaxial accelerometry on the wrist and waist, using the counts cut-off points for the accelerometers and ventilatory threshold one (VT1) and two (VT2). This research is a cross-sectional analytical study of university women (n = 35, 22.4 ± 1.9 years old). Triaxial accelerometers and portable ergospirometry were used during an incremental standardized walking-jogging test in an oval track of 160 m length (speed increments every minute with an audible signal from 0.84 m/s to 2.37 m/s). Accelerometry (from wrist and waist), oxygen consumption, and ventilatory response data were collected during the test. VT1 and VT2 were estimated through VSlope (VO2/VCO2) and carbon dioxide production/minute ventilation (VCO2/VE), respectively. An open-source statistical software was used. All the participants were able to finish the test. VT1 was found in stage 2 (n = 5), stage 3 (n = 5), stage 4 (n = 3), stage 5 (n = 8), stage 6 (n = 5), stage 7 (n = 3), stage 8 (n = 1), and in stage 9 (n = 5). VT2 was found during the incremental test only in 9 volunteers, in stage 7 (n = 2), in stage 8 (n = 3), stage 9 (n = 2), and stage 10 (n = 2). Differences between wrist and waist accelerometer measurements were observed in higher intensity stages of the test. Higher PA intensity measurements were found in the wrist, mainly above VT1. Also, there were differences between physical activity intensity levels detected by accelerometers and VT1 and VT2. The commonly used methods to assess physical activity intensity show no agreement in identifying the speed at which vigorous-intensity physical activity begins during an incremental test.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 220","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144585321","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"In vitro Reconstitution of Cytoskeletal Networks inside Phase Separated Giant Unilamellar Vesicles (GUVs).","authors":"Nishu Kanwa, María Reverte-López, Petra Schwille","doi":"10.3791/68530","DOIUrl":"https://doi.org/10.3791/68530","url":null,"abstract":"<p><p>Biomimetic lipid membranes in the form of giant unilamellar vesicles (GUVs) are commonly used to mimic cellular membrane behavior because of the ease of protein reconstitution inside GUVs, visualization, as well as understanding cellular membrane-protein dynamics. However, cell membranes comprise lipid rafts (or domains) arising from the presence of different lipids in the cellular membrane. Such increased complexity in model systems can be incorporated to result into phase separated GUVs, where lipid composition can be finely tuned. While encapsulation methods for the generation of homogeneous GUVs are widely known, methods to encapsulate proteins within phase separated GUVs are limited. Here, this protocol presents a simplified one-pot production of phase separated GUVs, comprised of liquid-disordered (Ld) and liquid ordered (Lo) domains, efficiently encapsulating different cytoskeletal proteins, i.e., FtsZ and actin, making the method a versatile tool for minimal cell production. Specifically, this approach uses an emulsion transfer protocol to produce GUVs with a high encapsulation efficiency. In this method, a lipid-monolayer is first generated by emulsifying a protein solution in a lipid/oil mixture, where lipids of varying phase transition temperatures are chosen to yield phase separation in the resultant GUVs. This emulsion is transferred gently on top of a lipid-in-oil solution in another tube, resulting in the formation of a water-oil interface. The solution is then centrifuged at elevated temperatures (ideally at 37 °C to retain protein activity), after which GUVs are collected for imaging. This method simplifies the in vitro reconstitution of cytoskeletal proteins within phase separated GUVs without using a cumbersome laboratory setup, and thus serves as a convenient method for studying the mechanics of cytoskeletal-membrane interactions in confinement.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 220","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144585345","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"In Vivo Imaging of Neural Activity in Unanesthetized Drosophila Adult Flies.","authors":"Prachi Shah, Isaac Cervantes-Sandoval","doi":"10.3791/68332","DOIUrl":"https://doi.org/10.3791/68332","url":null,"abstract":"<p><p>In vivo imaging is crucial for studying neurobiology in Drosophila as it allows real-time visualization of neuronal activity, development, and plasticity within the intact, behaving fly. This technique provides invaluable insights into dynamic processes, such as synaptic function and circuit connectivity, which cannot be accurately captured in fixed or ex vivo preparations. Most techniques for fly preparation for in vivo imaging involve anesthetizing the flies shortly before functional imaging. Nevertheless, extensive evidence demonstrates that anesthesia impacts several cognitive and physiological processes. For this reason, in vivo imaging of adult Drosophila melanogaster without the use of anesthetics during animal preparation is a challenging yet highly desirable approach. The protocol presented here involves immobilizing the head of awake flies by suctioning the proboscis into a small metal tube connected to a vacuum system. While the head is fixed, the fly is prepared for functional imaging without the need for anesthesia. This protocol is rapid and reproducible, ensuring no harm to the fly. The main advantage of this method lies in its avoidance of anesthetic use, thereby accounting for the potential complex effects of anesthesia on neural activity. This method is both affordable and highly adaptable, utilizing inexpensive, customizable tools. It allows for the successful imaging of live flies, capturing fast changes in neural activity associated with dynamic cognitive processes.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 220","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144585354","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}