Jove-Journal of Visualized Experiments最新文献

{"title":"Retrospective Evaluation of Synthetic Ligament-Based Revision of Anterior Cruciate Ligament Reconstruction for Postoperative Re-Rupture.","authors":"Qiang Ma, Li Shu, Aikeremujiang Muheremu","doi":"10.3791/68721","DOIUrl":"https://doi.org/10.3791/68721","url":null,"abstract":"<p><p>Re-rupture following anterior cruciate ligament (ACL) reconstruction remains a challenging complication, necessitating effective revision strategies. To evaluate the clinical and technical outcomes of synthetic ligament-based revision ACL reconstruction for postoperative re-rupture, a retrospective analysis was conducted on patients undergoing synthetic ligament revision ACL reconstruction between January 2023 and January 2024. Preoperative assessments included laboratory tests (ESR, CRP, tuberculosis antibodies), radiographic imaging (X-ray, CT, MRI), and functional scoring (VAS, IKDC, Lysholm). Surgical protocols emphasized meticulous tunnel planning, arthroscopic debridement, synthetic ligament placement, and dual fixation with metallic interference screws. Postoperative care involved early mobilization, systematic rehabilitation, and regular follow-ups. Results showed that the procedure demonstrated technical feasibility, with successful ligament fixation and no intraoperative complications. Postoperative evaluations revealed improved functional scores (IKDC, Lysholm) and reduced pain (VAS). Imaging confirmed proper ligament positioning and graft integrity. No instances of infection, graft failure, or significant joint instability were reported during follow-up. Those results indicate that synthetic ligament revision ACL reconstruction offers a reliable solution for re-rupture cases, combining precise surgical techniques with rigorous postoperative rehabilitation. This approach addresses anatomical challenges, restores knee stability, and enhances patient outcomes, supporting its adoption in complex revision scenarios.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 222","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144975360","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":"Classroom Behavior Recognition via Transformer and Attention Mechanism: Application in Teaching Quality Evaluation for AI Medical Education.","authors":"Lei Liu, Zhihua He","doi":"10.3791/69052","DOIUrl":"https://doi.org/10.3791/69052","url":null,"abstract":"<p><p>This study introduces a Transformer-based artificial intelligence system designed for classroom behavior recognition, aimed at enhancing teaching quality assessment and mental health monitoring in medical and nursing education. The model utilizes multimodal data, specifically video, audio, and skeletal movement, to evaluate key indicators of student engagement, such as attention span, interaction frequency, and emotional fluctuations. A novel multimodal fusion strategy, combined with spatiotemporal attention mechanisms, enables the system to capture complex classroom behaviors with improved precision and robustness. Experimental results on the EduSense and SBU Kinect datasets demonstrate that the proposed method significantly outperforms traditional models in both accuracy and false alarm rate. Additionally, ablation studies confirm the contribution of spatial and temporal attention modules to the system's recognition capability. The framework supports real-time feedback and visual behavior mapping, offering practical value in experiential learning environments. This approach provides a scalable and adaptive solution for classroom behavior monitoring and supports early intervention strategies in medical education.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 222","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144975275","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":"Establishment and Quantification of De Novo Lytic Infection by Cell-free Kaposi's Sarcoma-Associated Herpesvirus.","authors":"Qing Zhu, Janvhi Suresh Machhar, Pinghui Feng, Chengyu Liang","doi":"10.3791/68959","DOIUrl":"10.3791/68959","url":null,"abstract":"<p><p>Kaposi's sarcoma-associated herpesvirus (KSHV), a gammaherpesvirus implicated in multiple human malignancies, can undergo lytic replication during primary infection, a process that contributes to viral dissemination, immune evasion, and disease pathogenesis. However, the lack of robust in vitro systems for de novo lytic infection has limited insights into early infection events. Here, we present a tractable protocol that employs human colorectal cancer HCT 116 cells as targets for infection with cell-free virions derived from KSHV bacterial artificial chromosome 16 (BAC16)-reactivated iSLK producer cells. This model recapitulates key steps in primary infection, including viral entry, genome delivery, lytic gene expression, and progeny production. Infection is synchronized via spinoculation and monitored temporally. Virus production is quantified using a dual readout system comprising green fluorescence protein (GFP)-based infectious unit (IU) assays and qPCR of encapsidated viral DNA. These approaches enable detailed analysis of viral replication kinetics. While the current protocol focuses on the detection of late-stage events, the system is adaptable for studying early phases of infection through modified sampling time points. By offering a reproducible, scalable platform for productive de novo infection, this system addresses a methodological gap in KSHV research and supports mechanistic studies of herpesvirus-host interaction and antiviral strategies.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 222","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144975265","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 Gerbil Welfare Through Positive Reinforcement Training in Research Environments.","authors":"Dorothea Pichl, Paul Dechert, Anne-Maria Börner, Fernando Gonzalez-Uarquin, Sandra Reichel, Nadine Baumgart, Jan Baumgart","doi":"10.3791/68657","DOIUrl":"https://doi.org/10.3791/68657","url":null,"abstract":"<p><p>Improving laboratory animal welfare by minimizing stress and promoting species-appropriate handling is a central goal of contemporary biomedical research worldwide. Clicker training, a widely recognized form of positive reinforcement training, uses a click sound as a conditioned reinforcer to bridge the desired behavior and the reward, enabling animals to learn more quickly and with reduced stress. Our research group has previously demonstrated that clicker training functions as a form of cognitive enrichment in mice and rats. In the present study, we successfully adapted this training approach for Mongolian gerbils (Meriones unguiculatus), tailoring the protocol to the species-specific behavioral characteristics of gerbils. A cohort of 43 inbred gerbils (27 females and 16 males) underwent a standardized 10-day clicker training protocol, during which they learned to voluntarily approach and interact with the experimenter's hand in exchange for a food reward. Following the training period, animals were subjected to behavioral assessments, including an Open Field Test and a standardized human interaction test, to evaluate the effects of training on exploratory behavior and human-animal interaction. Our findings demonstrate that the implementation of clicker training in gerbils is fast, efficient, and well-tolerated. Trained animals, particularly females, showed increased voluntary interaction with the experimenter's hand and reduced anxiety-like behaviors. These results suggest that species-adapted clicker training protocols can facilitate the development of trust between experimenter and animal, ultimately decreasing stress and improving both animal welfare and the reliability of experimental outcomes.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 222","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144975255","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":"Method to Study Metabolism in Lymphoid Cells using Chemistry to Measure Puromycin Incorporation by Flow Cytometry.","authors":"Zenia Kaul, Hirofumi Shibata, Pamela L Schwartzberg","doi":"10.3791/67377","DOIUrl":"https://doi.org/10.3791/67377","url":null,"abstract":"<p><p>Upon antigen stimulation, naïve T cells undergo rapid proliferation and expansion to effector T cells. Metabolism plays an important role in the generation of biomass needed for these rapidly proliferating cells and for the generation of molecules required for effector T cell differentiation and function, which influence the outcome of the adaptive immune response in infection or cancers. Naïve T cells reprogram their metabolism upon antigenic stimulation to increase the generation of ATP, which is required to support their growth, biosynthesis, and effector functions. ATP can be generated in a cell either by the mitochondrial-oxidative phosphorylation (OXPHOS) pathway or by the glycolytic pathway. Because most of the ATP generated in a dividing, growing cell is used up for the synthesis of proteins, protein synthesis has been used as a surrogate for ATP levels. Protein synthesis can be measured by the incorporation of puromycin, which mimics the 3' adenosine of a tRNA charged with a modified tyrosine and leads to spontaneous termination of protein translation. Metabolic inhibitors like 2-deoxyglucose (2DG), which blocks the glycolytic pathway, and Oligomycin (O), which blocks complex 5 of the electron transport chain (ETC), can be used to study the dependencies of cellular ATP generation on these two pathways in conjunction with evaluation of protein synthesis in a method called SCENITH. We describe here a variation of this method that detects puromycin incorporation by flow cytometry using chemistry. This method of studying metabolism is relatively easy and can be used for evaluating rare cell populations, as well as patient samples, by flow cytometry.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 222","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144975270","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":"Comparative Study on the Polysaccharide Contents and Antioxidant Activities of Hippophae rhamnoides subsp. sinensis and Hippophae gyantsensis.","authors":"Chunqiao Shi, Yaning Lin, Guiying Ren, Yuying Song, Xing Yang, Jihang Xie, Yi Zhang, Yue Liu","doi":"10.3791/68855","DOIUrl":"https://doi.org/10.3791/68855","url":null,"abstract":"<p><p>This paper reports a well-designed and in-depth comparative study on the polysaccharide yields, contents, and antioxidant activities of two Hippophae species of great research value, namely, Hippophae rhamnoides subsp. sinensis Rousi and Hippophae gyantsensis (Rousi) Y. S. Lian. The total polysaccharides of H.rhamnoides subsp. sinensis and H. gyantsensis were obtained through hot water extraction. Yields were compared, and polysaccharide contents were accurately determined through phenol-sulfuric acid colorimetry and compared. Meanwhile, the antioxidant activity of the polysaccharides was evaluated through 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging. Results showed that the polysaccharide yield (percentage of crude extract relative to the raw material) of H.rhamnoides subsp. sinensis (1.18% ± 0.02%) was significantly lower than that of H. gyantsensis (3.12% ± 0.06%) (P < 0.01). The polysaccharide content (defined as pure polysaccharides in the extract) of H.rhamnoides subsp. sinensis (352.97±1.07 mg/g) was significantly higher than that of H. gyantsensis (300.21 ± 1.49 mg/g) (P < 0.01). The half-inhibitory concentration of the DPPH free radical scavenging activity of H.rhamnoides subsp. sinensis polysaccharides (0.026 ± 0.004 mg/mL) was slightly lower than that of H. gyantsensis polysaccharides (0.021 ± 0.004 mg/mL). However, no significant difference was found between the two Hippophae species (P > 0.05). This experiment clarified the differences in polysaccharide yield, content, and antioxidant activity between H.rhamnoides subsp. sinensis and H. gyantsensis. This work can provide a scientific basis for the quality evaluation of sea buckthorn and the further development of polysaccharide active components.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 222","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144975329","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 Prediction of 3D Domain Swapping Proteins in Medicinal Plants.","authors":"Aakanksha Pandey, Atul Kumar Upadhyay","doi":"10.3791/68519","DOIUrl":"https://doi.org/10.3791/68519","url":null,"abstract":"<p><p>3D domain swapping is a protein structural phenomenon in which two or more protein subunits exchange identical structural subunits and form oligomers. Proteins that exhibit 3D domain swapping play a crucial role in various biological functions, such as secondary metabolite biosynthesis, and in coping with several biotic and abiotic stresses in medicinal plants. This study investigates the ability to predict 3D domain swapping patterns among the genomes of medicinal plants using random forest and K-nearest neighbor classifiers models, demonstrating accuracies of 91.6% and 88.7%, respectively. A total of 420 (31%) of sequences were predicted as being putatively involved in 3D domain swapping. An enrichment investigation was also carried out on the predicted 3D domain-swapped protein sequences from various medicinal plants for function annotation based on Gene Ontology (GO) terms, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways analysis, and their domain distribution in secondary metabolites biosynthesis pathways. Functional annotation of predicted sequences infers that 3D domain swapped sequences were involved in diverse molecular functions such as photosynthetic electron transport in photo system II and electron transporters, transferring electrons within the cyclic electron transport pathway of photosynthesis activity, oxidative phosphorylation, and gene regulation of environmental stresses (biotic and abiotic) by synthesizing secondary metabolites (terpenoids, alkaloids, and polyamines). These findings underscore the ability of machine learning to predict the involvement of proteins in the 3D domain-swapping phenomenon, their respective function, and their potential to facilitate drug discovery and bioengineering initiatives.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 222","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144975315","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":"Surgical Technique for the Implantation of a Wireless Telemetry Device on Pigs' Backs.","authors":"Angel Moctezuma-Ramirez, Jordana Goulet, Abdelmotagaly Elgalad","doi":"10.3791/67810","DOIUrl":"https://doi.org/10.3791/67810","url":null,"abstract":"<p><p>Telemetry devices are essential for preclinical cardiovascular research, allowing for continuous monitoring of physiological parameters like electrocardiogram, temperature, and respiration in experimental animals without causing disruption. Traditional methods, such as intra-abdominal placement, can cause discomfort, particularly in pigs due to their habit of lying prone. To address this issue, a novel surgical technique was developed for implanting wireless telemetry devices in pigs, aimed at improving animal comfort and maintaining signal quality. This study involved 40 Yorkshire pigs (20-30 kg). The telemetry device was implanted in the back. A 2 inch incision was made lateral to the spine, followed by blunt dissection to create a pocket under the latissimus dorsi fascia. The pocket was treated with a vancomycin solution to prevent infection before device insertion. The leads were secured, and the site was closed in layers. All pigs showed strong, continuous telemetry signals after surgery. Over the 180-day follow-up period, only two cases of infection occurred: One was managed with antibiotics, and the other required device removal. No significant bleeding or loss of telemetry signal was observed, and all animals survived to the end of the study. This new technique offers a reliable method for implanting telemetry devices in pigs, reducing discomfort and infection risk while providing high-quality physiological data. It holds promise for broader use in preclinical cardiovascular research, improving both animal welfare and data reliability.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 222","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144975523","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":"A Simple Double Centrifugation Tube Method to Obtain Platelet-rich Plasma from Equine Blood.","authors":"Jorge U Carmona, Catalina López, Claudia Jurado-Grisales","doi":"10.3791/67985","DOIUrl":"https://doi.org/10.3791/67985","url":null,"abstract":"<p><p>Platelet-rich plasma (PRP) is widely utilized as a routine treatment for chronic musculoskeletal conditions in horses, such as osteoarthritis, tendinopathies, and desmopathies. This effectiveness stems from the high concentration of growth factors and anti-inflammatory cytokines that are released upon activation of this orthobiologic agent, whether activated endogenously or exogenously. Despite its growing popularity, there is a notable absence of instructional videos that demonstrate the techniques for obtaining PRP in horses. This study examines a double centrifugation tube method for obtaining PRP in horses. Whole blood from six horses was collected in sodium citrate tubes and centrifuged twice at 120 × g for 5 min and 240 × g for 5 min. The mean platelet concentration for PRP was 360.95 ± 56.31 platelets (PLT) × 10³/µL, while the mean leukocyte concentration was 3.05 ± 1.53 cells × 10³/µL. The platelet count in PRP was 2.41x higher than in whole blood, while the leukocyte concentration in PRP was 0.44x lower. The collection efficiency for PLT in PRP was 17%, while the collection efficiency for leukocytes in PRP was 3.1 %. The mean concentrations of transforming growth factor beta 1 and platelet-derived growth factor BB in PRP were 2601.82 (95% confidence interval (CI): 2358.88-2844.76) pg/mL and 1208.05 (95% CI: 1051.09-1365.02) pg/mL, respectively. These results indicate that PRP can be efficiently and reproducibly obtained through a simple, cost-effective method suitable for use by equine practitioners.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 222","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144975031","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":"Lateral Molar Approach-Driven Transoral Endoscopic Procedure for Benign Infratemporal Fossa Tumor Resection.","authors":"Shuai Chen, Bowen Yang, Xiaobo Dai, Guanru Wang, Xingzhi Zeng, Yonglin Song, Guiquan Zhu, Chunjie Li, Bing Yan","doi":"10.3791/68672","DOIUrl":"https://doi.org/10.3791/68672","url":null,"abstract":"<p><p>Benign infratemporal fossa tumors necessitate complete resection while preserving neurovascular integrity. Conventional open approaches risk delayed bone healing, occlusal dysfunction, severe facial scarring, and iatrogenic neurovascular injury. We propose an endoscopic-assisted plasma ablation technique via a lateral molar transoral approach to address these limitations. A 55-year-old female with a 2.5 × 2 cm right infratemporal fossa tumor underwent the novel procedure. A 4-5 cm mucosal incision lateral to the molar established an osteotomy-free corridor along the mandibular ramus. The technique integrated a 4-mm 0° rigid endoscope for magnified visualization and a low-temperature plasma ablation device for extracapsular dissection with simultaneous cutting/coagulation, minimizing thermal damage. Intraoperative neurophysiological monitoring safeguarded the mandibular nerve. Postoperative recovery metrics, MRI, and histopathology were analyzed. Complete tumor resection was achieved in 122 min with 50 mL blood loss. Postoperative MRI confirmed no residual lesions, but they were replaced by fibrotic scar tissue. Mandibular function (physiological mouth opening ≥35 mm) and facial sensation recovered by postoperative day 3. No hematoma, infection, or neurological deficits occurred. Histopathology confirmed a benign vascular malformation (SMA+, CD31/CD34+). Aesthetic outcomes were optimal, with no visible scarring (patient satisfaction: 10/10). The endoscopic-assisted plasma ablation technique via a transoral lateral molar approach enables precise, minimally invasive resection of anteromedial infratemporal fossa tumors (≤4 cm), avoiding osteotomy and external incisions. Synergizing anatomic landmark guidance, energy-selective ablation, and intraoperative neuroprotection, this method optimizes functional preservation and aesthetic outcomes while adhering to microsurgical principles. Standardization requires advanced endoscopic skills and warrants further validation in larger cohorts.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 222","pages":""},"PeriodicalIF":1.2,"publicationDate":"2025-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144975327","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}