Jove-Journal of Visualized Experiments最新文献

{"title":"Asymbiotic Germination and Leaf Explant-Based Regeneration of the Endangered Medicinal Orchid Hemipilia cucullata from Mature Seeds.","authors":"Yage Tu, Jianming Wang, Zhechen Qi, Weimei Jiang","doi":"10.3791/68541","DOIUrl":"https://doi.org/10.3791/68541","url":null,"abstract":"<p><p>Hemipilia cucullata (Orchidaceae) is a terrestrial orchid widely valued for its medicinal properties and ornamental appeal, yet its wild populations are endangered due to habitat degradation, low seed viability, and limited pollination success. To support its conservation and sustainable utilization, this study developed an optimized protocol for the asymbiotic germination of mature seeds, protocorm development, shoot elongation, and protocorm-like body (PLB) induction from sterile leaf explants. Seeds cultured in 1/2 MS liquid medium supplemented with 0.5 mg/L NAA showed a germination rate of 72 ± 6%, significantly higher than the control (46%) and solid medium (31%), with germination initiating approximately 21 days earlier in liquid culture. Protocorms were subcultured on B5 medium containing 0.5 mg/L BA, 0.2 mg/L NAA, and 100 mL/L coconut water, leading to efficient proliferation. After 5-6 weeks, plantlets were transferred to shoot elongation medium (B5 supplemented with 0.5 mg/L BA and 1 mg/L NAA), where they exhibited robust shoot elongation and healthy leaf formation. For PLB induction, the highest induction rate (44.3 ± 5.1%) was achieved on MS medium containing BA (3 mg/L) and NAA (0.2 mg/L). This comprehensive and reproducible protocol can be an effective platform for the large-scale propagation and ex situ conservation of H. cucullata, offering valuable support for recovery efforts of this endangered orchid.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 223","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145240103","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":"Methodology for the Study of Learning and Memory in Juvenile Rats Using Sevoflurane Anesthesia to Assess Hippocampal p-CREB and c-fos Expression.","authors":"Ye Jiang, Lai Jiang, Guohui Li","doi":"10.3791/68591","DOIUrl":"https://doi.org/10.3791/68591","url":null,"abstract":"<p><p>Studying cognitive impairments during brain development faces challenges due to the dynamic nature of neurodevelopment, methodological constraints, and contextual barriers. This study investigated an integrated methodological approach to study cognitive impairments in the developing brain caused by sevoflurane based on memory and p-CREB and c-fos expression in hippocampal neurons through the NMDAR/p-CREB/c-fos signaling pathway in juvenile rats. Sixty-four 21-day-old SD male rats (body weight <80 g) were divided into five groups: normal (n=8), 60% O2 (n=16), sham training (n=8), single-dose sevoflurane (n=16), and multiple-dose sevoflurane (n=16). The two sevoflurane groups were sub-grouped into sacrifice 1 day (groups sev1a and sev1b) and 30 days (groups sev30a and sev30b) after training (n=8/subgroup); all other rats were sacrificed 1 h after training. The Y-maze test was used to evaluate learning and memory retention. Immunohistochemistry was used to determine the number of p-CREB- and c-fos-positive neurons in the hippocampus specimens. Compared with the 60% O2 group (memory retention of 86.33% ± 17.52%), the memory of rats in the sev1a and sev1b groups showed a significant decrease (sev1a: 44.29% ± 11.26%; sev1b: 62.42% ± 7.27%; all P<0.05), while the sev1b and sev30b groups showed no significant differences (sev30a: 84.41% ± 14.15%; sev30b: 85.21% ± 11.61%; all P>0.05). Compared with the 60% O2 group (c-fos: 92.83 ± 7.88; p-CREB: 72.22 ± 8.89), the expression of p-CREB and c-fos in hippocampal neurons decreased in the sev1a group (c-fos: 23.13 ± 3.28; p-CREB: 22.88 ± 5.18) and the sev30 group (c-fos: 23.22 ± 3.13; p-CREB: 25.58 ± 2.26) (all P<0.05), while the sev30a and sev30b groups showed no significant differences (all P>0.05). Sevoflurane has a short-term negative stimulating effect on the memory maintenance ability of juvenile rats without an impact based on dosing frequency. Sevoflurane decreases the expression of p-CREB and c-fos in hippocampal neurons of juvenile rats.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 223","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145240140","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":"Enhancing Chimeric Antigen Receptor-Extracellular Vesicles (CAR-EV) Technology: The Future of Cancer Therapy.","authors":"Kartini Asari, Sharenya Chelvaretnam, Kol Thida Mom, Sadman Bhuiyan, Quang Pham, Amirah Fitri, Carlos Palma, Mozhgan Shojaee, Ramin Khanabdali, Leearne Hinch, Gregory Rice","doi":"10.3791/68726","DOIUrl":"https://doi.org/10.3791/68726","url":null,"abstract":"<p><p>CAR cell therapies have significantly advanced personalized treatment for several hematological malignancies. Currently, seven CAR- cell products are approved by the Food and Drug Administration (FDA) and six by the European Medicines Agency (EMA) for treating lymphoma, multiple myeloma, and chronic lymphocytic leukemia. Several challenges and limitations remain, with cytokine release syndrome (CRS) and immune effector cell-associated neurotoxicity syndrome (ICANS) being the most significant. Cell-free therapies, such as CAR-EVs, offer substantive advantages over their cellular counterparts. These include enhanced tumor infiltration and the potential for repeat administration while minimizing the risks of CRS, ICANS, and other adverse side effects. Additionally, the potency of CAR-EVs can be tuned by engineering the inclusion of cytotoxic agents and function-modifying ribonucleic acids (RNAs). Herein, we report on the development of a scalable CAR-EV platform for producing tunable CAR-EVs. This platform includes the engineering and pre-conditioning of EV producer cells (e.g., CAR-T and CAR-natural killer (CAR-NK) cells), isolation and enrichment of CAR-EVs using a Good Manufacturing Practice (GMP) grade ion-exchange chromatography (IEX) platform, fully automated high-throughput EV subpopulation analysis, and in vitro evaluation of CAR-EV functional cytotoxic activity. The platform has been validated using CAR-NK-EVs and CAR-T-EVs for both hematological and solid tumor cell lines. The CAR-EV platform represents a promising approach for the rapid development of off-the-shelf therapeutic CAR-EVs tailored to specific disease indications, with the potential to reduce adverse side effects associated with CAR-cell-based therapies.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 223","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145240087","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":"Machine Learning-Based Cough Tone Classification: Diagnostic Exploration of Chronic Obstructive Pulmonary Disease and Respiratory Tract Infections.","authors":"Xiaohan Liu, Tengteng Li, Jingxin Zhang, Jiutang Sun, Jinghua Li, Zhulv Zhang, Benzhang Zhao, Jianjun Wu, Yu Lu, Tao Lu","doi":"10.3791/68222","DOIUrl":"https://doi.org/10.3791/68222","url":null,"abstract":"<p><p>The objective of this study was to develop and evaluate a non-invasive method for distinguishing patients with chronic obstructive pulmonary disease (COPD) from those with respiratory tract infections (RTI) using voice signal analysis and machine learning. Fixed-pattern voice signals were collected from 25 COPD patients and 25 RTI patients (serving as the control/comparison group). Multi-dimensional voice feature analysis was performed to identify features significantly differentiating the two groups. Statistically significant features were selected and subjected to dimensionality reduction. Logistic Regression (LR) and Random Forest (RF) models were then trained and evaluated for classification performance in distinguishing COPD from RTI. Over 400 voice features were initially analyzed. Eighteen features showed highly significant differences between COPD and RTI patients (P <; 0.05). In the task of distinguishing COPD patients from RTI patients, the LR model achieved a test set area under the curve AUC of 0.95, significantly outperforming the RF model (AUC = 0.76). This study demonstrates the feasibility of using voice analysis and machine learning, particularly the LR model, as a promising non-invasive tool for differentiating COPD from RTI. It provides a foundation for the practical application and further optimization of this voice-based approach in clinical settings requiring differential diagnosis of respiratory conditions.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 223","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145240123","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":"Automating Tumor Implantation in Zebrafish Larvae for Cancer Research and Medicine.","authors":"Wietske van der Ent, Yi Ding, Michael Jury, Michele Scotto Di Mase, Kees-Jan van der Kolk, Camila Vincencio Esquerra, Jan de Sonneville, Lasse D Jensen","doi":"10.3791/68441","DOIUrl":"https://doi.org/10.3791/68441","url":null,"abstract":"<p><p>Zebrafish larval injections, including subcutaneous and brain-orthotopic tumor xenografting as well as intravenous drug administration, have become central techniques in zebrafish-based research. These procedures, however, remain tedious, technically challenging, and highly dependent on operator skill, limiting throughput and reproducibility. To address these limitations, we introduce an advanced robotic system that automates microinjections into zebrafish larvae, significantly improving efficiency, precision, and reproducibility over manual injections. Here, we provide a detailed protocol for operating the automated injector, a high-throughput system capable of delivering liquids, particles, or cancer cells at three different anatomical sites with superior accuracy and speed compared to trained zebrafish researchers. We outline the essential steps, including setup, calibrations, injection procedures, and troubleshooting, to ensure optimal performance. Furthermore, we provide representative examples of results from using the robot and discuss its potential to facilitate large-scale larval injections, including for drug screens and precision medicine. By integrating an automated injector into the workflows, researchers can overcome limitations of manual injections and accelerate preclinical drug discovery and personalized treatment selection.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 223","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145240047","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":"Application of Optogenetics Combined with Fiber Photometry Recording in the Regulation of Neural Circuits-visceral Functions by Electroacupuncture.","authors":"Yueyue Liu, Yun Liu, Ying Wang, Zhu Bing, Xinyan Gao","doi":"10.3791/68667","DOIUrl":"https://doi.org/10.3791/68667","url":null,"abstract":"<p><p>This protocol describes a technique for optogenetic manipulation of specific types of neurons and real-time recording of the effects of electroacupuncture at Zusanli (ST36) on nuclear calcium signals and gastric motility in mice under anesthesia in vivo. The exploration of the central mechanisms underlying the efficacy of acupuncture has been limited by technological constraints. However, the advancement of optogenetic and optical imaging technology has propelled the development of neuroscience. Fiber photometry recording, initially leveraging genetically encoded calcium indicators to visualize changes in calcium dynamics indicative of neuronal activity, stands out as a key technology for characterizing brain-behavior correlations in vivo. The application of optogenetics enables the genetic encoding of neurons, enabling their activation or inhibition in response to light stimulation. This capability facilitates the establishment of causal links between neural circuitry function and behavioral outcomes. Simultaneous fiber photometry, calcium recording, and optogenetic stimulation provide a method for real-time recording and manipulation of neuronal activity, serving as an effective approach to investigate the central mechanisms of acupuncture. Nonetheless, there remains a scarcity of studies documenting the concurrent utilization of fiber photometry calcium recording, optogenetics, and visceral function monitoring in acupuncture research. The results of this experiment show that EA-ST36 might regulate gastric motility related to PBN^Glu-NTS^Glu-DMV^ChAT circuit. Optogenetic stimulation of this circuit produces the same effect. After the superposition of the two, the changes in calcium signal and gastric motility reach their maximum. This study established a novel approach for the simultaneous integration of optogenetic, intracerebral calcium signal recording, and gastric motility monitoring, offering a novel paradigm for real-time monitoring of the relationship between somatic stimulation and neural circuit-visceral function interactions.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 223","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145240089","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":"Optimized Analysis of Proteins from Xenopus Oocytes and Embryos by Immunoblotting.","authors":"Charlotte R Kanzler, Michael D Sheets","doi":"10.3791/69139","DOIUrl":"https://doi.org/10.3791/69139","url":null,"abstract":"<p><p>Analysis of proteins by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by immunoblotting (western blotting) is a vital part of the molecular biologist's toolkit. This technique separates a complex protein mixture by molecular weight and then assays the presence of target proteins using specific antibodies. Immunoblotting has a variety of applications. Examples include use as a targeted approach to study protein-protein interactors or as a control to confirm expression or depletion of protein targets. However, the successful execution of immunoblotting requires complicated, multistep experiments. Protocols must be optimized for each organism, target protein, and application. Therefore, knowledge gaps exist for the use of immunoblots in many models, including the model frog Xenopus laevis. Due to their large size, abundant material for biochemical experiments, and facile handling, X. laevis oocytes and embryos have been vital for studying principles of translational control. However, this species lacks specific protocols for robust and routine immunoblotting. Here, we offer an in-depth protocol for western blotting optimized for samples from multiple Xenopus developmental stages. We then analyze translational regulators across development.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 223","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145240160","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":"Standardized Diagnosis and Treatment of Pancreaticobiliary Maljunction.","authors":"Yongcong Yan, Haohan Liu, Zhiyu Xiao, Chuanchao He","doi":"10.3791/69050","DOIUrl":"https://doi.org/10.3791/69050","url":null,"abstract":"<p><p>According to extensive case analyses compiled by the Japanese Study Group on Pancreaticobiliary Maljunction (JSPBM), patients with Pancreaticobiliary Maljunction (PBM) exhibit a significantly elevated incidence of biliary tract tumors, ranging from 34% to 39%. This rate is dramatically higher than the population-based incidence of biliary tract tumors in the general population (approximately 0.002%). Beyond malignancy, PBM is associated with multiple common pancreatobiliary disorders, including recurrent pancreatitis, stones within the common pancreatobiliary channel, and pancreatic duct stones, which substantially impair patients' quality of life. Despite its insidious nature and frequent delayed or missed diagnosis, key clinical features such as recurrent pancreatitis, acalculous cholecystitis, and extrahepatic bile duct dilation provide crucial diagnostic clues. Standard diagnostic modalities include abdominal ultrasound, computed tomography (CT), and magnetic resonance cholangiopancreatography (MRCP). Endoscopic retrograde cholangiopancreatography (ERCP) serves as the definitive diagnostic and classification tool. Upon confirmation of diagnosis, patients should undergo curative surgery to mitigate the risk of biliary tract carcinogenesis. This article provides a detailed protocol for the diagnostic and therapeutic approaches to PBM.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 223","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145240138","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":"Revealing Electromechanical Control of Tissue Homeostasis Using a Two-Layer Microfluidic Device.","authors":"Xiaolu Jiang, Ping Xu, Feng Feng, Gianluca Grenci, Thuan Beng Saw","doi":"10.3791/68894","DOIUrl":"https://doi.org/10.3791/68894","url":null,"abstract":"<p><p>Cell behavior and cell fate are impacted by electric currents or fields that are endogenous or externally applied. Static electric signals can be applied in customized microfluidic devices to mimic the electric environment in slow physiological processes such as development, wound healing, and homeostasis. An important class of cellular electric studies is the control of cell migration by static in-plane galvanic currents in simple microfluidic channels mimicking wound currents, with current densities of ~0.1-1000 A/m². However, due to incompatible geometry, these devices are not appropriate to study electric effects in tissue homeostasis, where cells adopt apico-basal polarity and a transepithelial potential difference (TEPD). Here, we detail a unique microfluidic-based device that applies physiological ion currents perpendicular to the plane of confluent epithelial cell layers to perturb the TEPD and investigate electrical regulation of tissue steady states. The setup is made from a two-layer UV-curable polymer embedded with soft, polyacrylamide gel substrate coated with extracellular-matrix protein of choice. This microfluidic device provides the correct geometry and permeable substrate to induce a relatively uniform ion current across the cell layers of centimetric-scale. The setup is compatible with confocal live-cell imaging and Traction Force Microscopy to infer mechanical stresses induced by the transepithelial currents. Strikingly, the proliferation, extrusion and migration of cells are collectively influenced within the confluent epithelium depending on the direction of ion current, inducing a new tissue state characterized by different cell-cell interaction strengths, cell events (death and proliferation), and tissue structures. The electrically controlled cell behaviors can be understood as an electrically induced mechanical stress and cell response. This novel microfluidic device and protocol provide the tool and documentation required for the mechanobiology and bioengineering communities to study electric effects in tissue homeostasis and develop novel tissue engineering applications.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 223","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145240180","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":"TD-DFT Guided Advanced E-Eye Sensing Technique for On-site Quantification of Fe, Cr, F, and As in the Environmental, Biological, and Food Samples.","authors":"Shrikant Kashyap, Rajasekhar Ravula, Neha Majee, Tapas K Mandal","doi":"10.3791/68767","DOIUrl":"https://doi.org/10.3791/68767","url":null,"abstract":"<p><p>This paper demonstrates the step-by-step protocol of designing and developing an E-Eye-enabled multiplexed point-of-care testing (POCT) kit. We use time-dependent density function theory (TD-DFT) to explore the basic working principle of the sensors before going into the details of the device's hardware. The TD-DFT analysis gives the λmax of the targeted element, which helps find the most accurate route for detecting the pollutant in the analytes. The TD-DFT analysis is performed in Gaussian 09 and Gauss View 5.0 software. An optical sensor called the electronic eye (E-Eye) has been developed based on the λmax value and the Light Emitting Diode/Light Dependent Resistor (LED/LDR) principle. The E-Eye device has been fabricated with an LED, an LDR placed opposite each other, and other hardware, including a liquid crystal display (LCD) interfaced with a microprocessor across a voltage divider, acting as a central microprocessor. Initially, all the samples (environmental, biological, and food and beverages) were pre-processed to extract all the targeted elements in the aqueous phase. The specific (lock and key) reaction has been carried out in the specified reactor, and readings observed in the display unit have been recorded. An indigenously multiplexed device has also been fabricated to detect Fe, Cr, As, and F simultaneously. The efficacy of the sensors has been tested against more than 2000 samples and compared with the gold standard methods. A good precision has been confirmed with the accuracy of 95.3%, 94.7% and 95.4% with respect to the samples of environmental, biological, and food and beverages. It is important to note that the performance of the arsenic sensor has been compared with Atomic Absorption Spectrometry (AAS) results, and all other results, i.e., sensors for iron, chromium, and fluoride, have been compared with the results obtained from UV-Vis analysis.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 223","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145240197","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}