Jove-Journal of Visualized Experiments最新文献

{"title":"Matrix-based DNA Extraction for Targeted Next-Generation Sequencing on Decontaminated Sputum Samples.","authors":"Jennifer Williams, Janré Steyn, Emilyn Costa Conceição, Felicia Bernita Wells, Melanie Grobbelaar, Nabila Ismail, Yonas Ghebrekristos, Christoffel Johannes Opperman, Sarishna Singh, Jason Limberis, Fahd Naufal, Brendon Coenrad Mann, Rebecca E Colman, Timothy Rodwell, Robin Mark Warren, John Z Metcalfe","doi":"10.3791/68147","DOIUrl":"https://doi.org/10.3791/68147","url":null,"abstract":"<p><p>Next-generation sequencing (NGS) is now recognized as a powerful tool for timely and accurate drug-resistant tuberculosis (DR-TB) diagnosis. Targeted NGS (tNGS) offers a streamlined approach by focusing on specific genes associated with drug resistance, bypassing the need for traditional culture-based methods with turnaround times ranging from weeks to months. The World Health Organization (WHO) has recommended tNGS as a valuable strategy for improving tuberculosis (TB) diagnosis to guide treatment and improve treatment outcomes, particularly in resource-limited settings. Among the WHO-recommended tNGS assays, we have selected a method that provides rapid and comprehensive drug susceptibility testing, lineage determination, and strain typing. While standardized DNA extraction methods are available, they can be time-consuming and labor-intensive. To address this challenge, we optimized a simplified, matrix-based DNA extraction protocol in combination with magnetic bead purification. This method offers a rapid and efficient approach for extracting DNA directly from decontaminated sputum sediments, enabling rapid downstream tNGS analysis. By streamlining the DNA extraction process from sputum sediment, this protocol could facilitate wider adoption of tNGS in routine clinical settings, ultimately contributing to improved patient outcomes and lending to global TB control efforts.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 220","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144477488","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":"Treatment with Locking Intramedullary Nailing for Intertrochanteric Fracture of the Femur Utilizing a New Awl with a Distal Positioner.","authors":"Zongpei Lian, Binbin Liu, Zehang Zheng, Chuang Huang, Fei Xu, Yang Liu, Zhengqiang Luo","doi":"10.3791/64841","DOIUrl":"10.3791/64841","url":null,"abstract":"<p><p>Interlocked intramedullary nailing is now established as the preferred method of managing femoral intertrochanteric fractures. Choosing the ideal entry point and inserting the guidewire accurately are key steps of the procedure. Several factors make the procedure more challenging, such as the supine position, obese patients, and the tip of the trochanter not aligning co-linear with the anatomic axis of the femoral medullary canal in the frontal plane. Our team has developed a new awl with a distal positioner that assists the guidewire insertion and entry portal of femurs. This comparative study analyzed 40 intertrochanteric fracture cases treated with locking intramedullary nailing, which were randomized to receive either the new awl incorporating a distal positioner (n = 20) or the conventional guide apparatus (n = 20). Operation time, blood loss, the success rate of the one-time insertion, radiation times, and bone healing time were recorded. The patients (21 males and 19 females) were treated with Gamma 3 and proximal femur nail antirotation (PFNA) with the help of the new awl with a distal positioner or conventional guide apparatus. There were no transoperative complications in the two groups. The surgical time in the new guide apparatus group was significantly shorter compared to the control group. The new awl group achieved a 100% success rate for one-time needle insertion, surpassing the control group's rate of 66.7%. The fluoroscopy time required for the new awl group was obviously shorter compared to that of the control group. However, no significant differences were observed in terms of intraoperative blood loss or bone healing time between the two groups. The newly designed guide awl with a distal positioner could reduce the difficulty in opening the femur for inserting the interlocked intramedullary nail. This tool is especially suitable for obese patients.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 220","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144477453","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":"Using Computer Vision Libraries to Streamline Nuclei Quantification.","authors":"Danielle E Levitt, Alexandra L Khartabil, Rylea E Hall, Matthew R DiLeo, Connor J Mills, Ashley K Williams, Casey R Appell, Ronald G Budnar, Hui-Ying Luk","doi":"10.3791/67945","DOIUrl":"https://doi.org/10.3791/67945","url":null,"abstract":"<p><p>Live cell assays and image-based cell analyses require data normalization for accurate interpretation. A commonly used method is to stain and quantify nuclei, followed by data normalization to nuclei count. This nuclei count is often expressed as cell count for uninucleate cells. While manual quantification can be laborious and time-consuming, available automated methods may not be preferred by all users, may lack validation for this specific application, or may be cost-prohibitive. Here, we provide step-by-step instructions for capturing quantifiable images of nuclei stained with fluorescent DNA stains and subsequently quantifying the nuclei using an automated object counting software program developed using Python computer vision libraries. We also validate this program across a range of cell densities. Although the exact time for program execution varies based on the number of images and computer hardware, this program consolidates hours of work counting nuclei into seconds for the program to run. While this protocol was developed using images of fixed, stained cells, images of stained nuclei in live cells and immunofluorescence applications can also be quantified using this program. Ultimately, this program provides an option that does not require a high degree of technological skill and is a validated, open-source alternative to aid cell and molecular biologists in streamlining their workflows, automating the tedious and time-consuming task of nuclei quantification.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 220","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144477456","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":"Separation and Differential Characterization of Gut Microbial Extracellular Vesicles in Salt-Sensitive Rats under High-Salt Diet Conditions.","authors":"Lisha Wang, Lijuan Cui, Luming Qi, Youli Tan, Shuyu Wang, Hongling Liu, Changhong Wei, Zhangmeng Xu, Jie Wang","doi":"10.3791/68279","DOIUrl":"https://doi.org/10.3791/68279","url":null,"abstract":"<p><p>High salt intake is a major risk factor for hypertension, and its underlying mechanism may be closely linked to extracellular vesicles (EVs) secreted by gut microbiota. These EVs, produced by gut microbiota, carry various bioactive components that may play a crucial role in the development of hypertension induced by a high-salt diet (HSD). To investigate this mechanism, we developed an efficient extraction method based on density gradient centrifugation to isolate EVs from the gut microbiota of salt-sensitive rats fed an HSD. Through particle size analysis, transmission electron microscopy (TEM), and lipopolysaccharide (LPS) detection, we identified the gradient distribution of gut microbiota EVs and achieved precise extraction. Furthermore, 16S rRNA gene sequencing was employed to analyze the origin and compositional differences of EVs between the normal and HSD groups, revealing the impact of high salt intake on the genetic characteristics of gut microbiota EVs. This study provides valuable tools and scientific insights into the gut microbiota mechanisms underlying salt-induced hypertension and offers new perspectives for the prevention and treatment of related diseases.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 220","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144477507","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":"Intracellular Phosphoflow Cytometry of Acute Myeloid Leukemia Patient-Derived Xenotransplants.","authors":"Victor Gife, Bahram Sharif-Askari, Anavasadat Sadr Hashemi Nejad, Raquel Aloyz, Laura Hulea, François E Mercier","doi":"10.3791/68244","DOIUrl":"10.3791/68244","url":null,"abstract":"<p><p>To adapt and resist approved treatments, acute myeloid leukemia (AML) cells activate specific molecular pathways that lead to changes in gene expression, protein levels and activity. In this protocol, an approach is reported to explore targets phosphorylated downstream of oncogenic signaling in AML: p-STAT5 (Tyr694), p-4EBP1 (Thr37/46), p-RPS6 (Ser240/244), and p-ERK1/2 (Thr202/Tyr204). This method enables the assessment of how these pathways-major regulators of stemness maintenance, immune evasion, protein synthesis, and adaptation to oxidative and metabolic stress-are modulated by one or more tested compounds in bone marrow cells harvested from live mice by aspiration before and after the treatment phase. This minimally invasive method preserves cell integrity and reduces stress compared to bone-crushing techniques, which can induce damage and potentially affect experimental outcomes. To optimize intracellular antibody staining for flow cytometric analysis, a protocol was developed using paraformaldehyde fixation and methanol permeabilization. This approach ensures high staining precision and minimizes background noise, enabling reliable detection of intracellular signaling markers. One of the main advantages of this protocol is the development of a multiparametric antibody panel, allowing for simultaneous assessment of the four pathways within the same sample. Using a next-generation spectral flow cytometer with high sensitivity, dynamic shifts in pathway activation were observed depending on treatment conditions compared to pretreatment baseline levels in the same mice. This methodology enables precise in vivo analysis of signaling pathway modulation in patient-derived xenograft bone marrow samples without requiring euthanasia of the animals, providing valuable insight into the adaptive mechanisms of AML cells, and can guide the evaluation of therapeutic strategies aimed at targeting these pathways to overcome resistance.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 220","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144477486","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":"Transperineal Prostate Biopsy Using a Cone-shaped Double-hole Method with Dual-plane Probe Guidance.","authors":"Gang Fu, Mingyi Shui, Xiaoting Wang","doi":"10.3791/68038","DOIUrl":"10.3791/68038","url":null,"abstract":"<p><p>Prostate cancer, the second most common malignancy in men, frequently necessitates biopsy for accurate diagnosis. This study presents a transperineal cone-shaped double-hole prostate biopsy method guided by micro-ultrasound, performed under local anesthesia. This innovative technique offers a minimally invasive approach that maintains high diagnostic accuracy while enhancing patient comfort and reducing adverse events. The cone-shaped double-hole method is designed to minimize tissue trauma and reduce recovery times. The dual-plane ultrasound probe facilitates precise, real-time needle navigation and improves lesion-targeting accuracy. Our retrospective analysis of 526 cases demonstrated that the improved method significantly reduced the procedure time for lesion localization from 30 min to 10 min using dual-plane probe guidance. Furthermore, statistical comparisons revealed a marked reduction in pain scores, and notably, the infection rate has remained at zero to date. Compared to previous techniques that required general or spinal anesthesia, this method minimizes trauma and expedites recovery, with only two small needle punctures visible on the skin post-procedure. The findings underline the efficacy and safety of this simple, minimally invasive prostate biopsy technique under micro-ultrasound guidance, offering an alternative that improves patient outcomes without compromising diagnostic accuracy.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 220","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144477511","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":"Quantification of Polybutylene Adipate Terephthalate-based Micro- and Nano-plastics from Soil Using Proton Nuclear Magnetic Resonance Spectroscopy.","authors":"Alexis B Gillmore, Carlos A Steren, Douglas G Hayes, Sean M Schaeffer","doi":"10.3791/67471","DOIUrl":"10.3791/67471","url":null,"abstract":"<p><p>A method to recover and quantify micro- and nano-plastics (MPs and NPs) formed in the soil during biodegradation is needed to accurately assess the degradation and environmental impact of biodegradable plastic products. The presence of MPs and NPs in soil may alter soil properties like aggregation behavior or have toxic effects on soil biota. Existing MP recovery methods are not always suitable for measuring biodegradable polymers like polybutylene adipate terephthalate (PBAT); some common digestion procedures with acids or oxidizers can destroy PBAT-based biodegradable MPs. Identification methods like micro-FTIR and micro-Raman spectroscopy are also limited by the minimum size of particles that can be recovered and analyzed. Therefore, this method was developed to extract and quantify PBAT from soil to assess the mass fraction of MPs and NPs in the soil without chemically transforming PBAT. In the protocol, a chloroform-methanol solution is used to selectively extract PBAT from the soil. The solvent is evaporated from the extract, and then the extract is redissolved in deuterated chloroform. The extract is analyzed by proton nuclear magnetic resonance spectroscopy (¹H-QNMR) under quantitative parameters to quantify the amount of PBAT in each sample. Solvent extraction efficiencies for PBAT range from 76% in a shady loam soil to 45% in an Elkhorn sandy loam soil. PBAT recovery may be reduced for photo-oxidized materials compared to pristine ones and may be reduced in soils with high clay content. Extraction efficiencies do not depend on PBAT concentration within the test range, but lower extraction efficiencies were observed for NPs than for MPs. PBAT quantification results were comparable to the quantification of plastic degradation by measuring cumulative soil respiration in a laboratory incubation study.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 220","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144477504","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":"Randomized, Triple-Blind, and Parallel-Controlled Trial of Transcranial Direct Current Stimulation for Cognitive Rehabilitation after Stroke.","authors":"Begoña González Rodríguez, Elena M Marrón, Marcos Ríos-Lago, Aida Arroyo Ferrer, Yeray González-Zamorano, Francisco José Sánchez Cuesta, David De Noreña, Juan Pablo Romero","doi":"10.3791/67809","DOIUrl":"https://doi.org/10.3791/67809","url":null,"abstract":"<p><p>Right hemisphere stroke frequently results in hemispatial neglect, a disabling condition that can significantly impede the recovery process. The chronic presence of neglect has been linked to poorer outcomes in both cognitive and motor domains. As an adjunct to conventional neuropsychological interventions, transcranial direct current stimulation (tDCS)-a noninvasive technique that modulates neural excitability through low-intensity electrical currents-has gained attention for its potential to enhance cortical plasticity and support functional improvement in affected individuals. In this study, we propose a combined intervention protocol aimed at reducing symptoms of post-stroke hemispatial neglect. It consists of a cathodal tDCS protocol combined with a computerized neuropsychological rehabilitation program specifically designed for the rehabilitation of hemispatial neglect. The neuromodulation strategy is to reduce the hyperactivation of the undamaged hemisphere based on the interhemispheric rivalry model. The intervention consists of 2 weeks, 10 sessions (from Monday to Friday), 45 min each, of tDCS and conventional cognitive stimulation concurrently applied. The tDCS is applied by an 8-channel high definitiontDCS (HD- tDCS) device for 20 min and at 2 mA of intensity. The cathode is positioned over the left posterior parietal cortex (P3 following 10/20 system for electroencephalogram [EEG] electrode placement), and return electrodes are placed at C3, CP5, CP1, Pz, PO3, PO7, and P7. A neuropsychological and functional assessment was carried out at baseline and after the end of the intervention. The primary aim of the present study is to describe the protocol for a parallel, randomized, triple-blind experimental design. To ensure the feasibility of the protocol and its potential efficacy, a comprehensive description of the procedures applied to a single pilot participant is provided. Incorporating tDCS neuromodulation strategies into cognitive rehabilitation processes may lead to shortened intervention times and improve the functional status and quality of life of patients.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 220","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144477505","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":"Chronic Cranial Window Technique for Repeated Cortical Recordings During Anesthesia in Pigs.","authors":"Suzan Meijs, Felipe R Andreis, Benedict Kjærgaard, Taha A M Janjua, Winnie Jensen","doi":"10.3791/67931","DOIUrl":"https://doi.org/10.3791/67931","url":null,"abstract":"<p><p>Cortical recordings are essential for extracting neuronal signals to inform various applications, including brain-computer interfaces and disease diagnostics. Each application places specific requirements on the recording technique, and invasive solutions are often selected for long-term recordings. However, invasive recording methods are challenged by device failure and adverse tissue responses, which compromise long-term signal quality. To improve the reliability and quality of chronic cortical recordings while minimizing risks related to device failure and tissue reactions, we developed a cranial window technique. In this protocol, we report methods to implant and access a cranial window in juvenile landrace pigs, which facilitates temporary electrocorticography (ECoG) array placement on the dura mater. We further describe how cortical signals can be recorded using the cranial window technique. Cranial window access can be repeated several times, but a minimum of 2 weeks between implant and access surgeries is advised to facilitate recovery and tissue healing. The cranial window approach successfully minimized common electrode failure modes and tissue responses, resulting in stable and reliable cortical recordings over time. We recorded event-related potentials (ERPs) from the primary somatosensory cortex as an example. The method provided highly reliable recordings, which also allowed the assessment of the effect of an intervention (high-frequency stimulation) on the ERPs. The absence of significant device failures and the reduced number of electrodes used (two electrodes, 43 recording sessions, 16 animals) suggest an improved research economy. While minor surgical access is required for electrode placement, the method offers advantages such as reduced infection risk and improved animal welfare. This study presents a scalable, reliable, and reproducible method for chronic cortical recordings, with potential applications in various fields of neuroscience, including pain research and neurological disease diagnosis. Future adaptations may extend its use to other species and recording modalities, such as intracortical recordings and imaging techniques.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 220","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144477466","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":"Generation and Characterization of Human Induced Pluripotent Stem Cell-derived Astrocytes Lacking Fragile X Messenger Ribonucleoprotein.","authors":"Bharath Kumar Reddy, Nikhita Annaiyappa, Aditi Bhattacharya, Sumantra Chattarji, Rakhi Pal","doi":"10.3791/68081","DOIUrl":"https://doi.org/10.3791/68081","url":null,"abstract":"<p><p>Fragile X syndrome (FXS), a leading inherited cause of autism spectrum disorder and intellectual disability, has been studied extensively using rodent models. More recently, human stem cell-derived model systems have also been used to gain mechanistic insights into the pathophysiology of FXS. However, these studies have focused almost exclusively on neurons. Further, despite growing evidence for a key role of glia in neuronal function in health and disease, little is known about how human astrocytes are affected by FXS. Therefore, in this study, we successfully developed a protocol that captures key spatiotemporal milestones of brain development and aligns with the process of gliogenesis as well. Together this offers a useful framework for studying neurodevelopmental disorders. First, we patterned the human induced pluripotent stem cells into the neuroectodermal lineage with dual Suppressor of Mothers against Decapentaplegic (SMAD) inhibition and small molecules. Subsequently, we utilized specific growth factors and cytokines to generate control (CTRL) and FXS patient-derived astrocytic progenitor cells (APCs). Treatment of APCs with ciliary neurotrophic factor, a differentiating cytokine, regulated and drove the progenitor cells towards astrocytic maturation, yielding forebrain-specific glial fibrillary acidic protein-expressing astrocytes. We found that these astrocytes are functional, as evidenced by their calcium responses to ATP application, and they exhibit dysregulated glycolytic and mitochondrial metabolism in FXS. Taken together, these findings provide a useful experimental platform of human origin for the investigation of cell-autonomous and non-cell-autonomous consequences of alterations in astrocytic function caused by neurodevelopmental disorders.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 220","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144477482","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}