Jove-Journal of Visualized Experiments最新文献

Post-stroke depression (PSD) is a common, treatable complication of stroke, characterized by depressive and somatic symptoms that impair patient recovery and quality of life. This study describes a protocol combining middle cerebral artery occlusion (MCAO) with behavioral restraint and isolation housing to establish a rat PSD model, and evaluates the therapeutic effect of Yi-nao-jie-yu Prescription (YNJYP) on adult neurogenesis in PSD rats. For model establishment, rats in the stroke, PSD, fluoxetine hydrochloride (FXT), and YNJYP groups underwent MCAO: a monofilament suture was advanced from the internal carotid artery to the middle cerebral artery (MCA) for 2 h of ischemia, followed by reperfusion. From post-MCAO day 7, rats in the PSD, FXT, and YNJYP groups were single-housed and restrained in a custom T-shaped platform for 2 h daily for 7 days. Behavioral assessments included the forced swim test (FST, for despair), sucrose consumption test (SCT, for anhedonia), and open-field test (OFT, for exploratory behavior). At 4 and 8 weeks post-stroke, PSD rats showed longer immobility time in FST and lower sucrose preference in SCT than stroke rats (P < 0.01). YNJYP reversed these depressive-like behaviors (P < 0.01), with efficacy comparable to FXT. This protocol confirms the validity of the PSD model and YNJYP's therapeutic potential, supported by rigorous experiments and data analysis.

Measuring the viral titer, the concentration of a virus in a sample, is a fundamental procedure in virology research. While essential, traditional methods like the plaque assay and TCID50 assay can be time-consuming, require staining or labeling reagents, and often involve subjective interpretation, particularly when cytopathic effects (CPE) are subtle or difficult to quantify through imaging. For example, TCID50 assays may employ viability dyes like MTT or MTS, while plaque assays rely on imaging and staining to visualize viral plaques, both of which can introduce variability. Moreover, these traditional methods only offer a static snapshot of viral infectivity, limiting the ability to capture the dynamic interactions between viruses and permissive cells. To overcome the current limitations in measuring virus titer, a streamlined TCID50 assay was developed using impedance-based technology to objectively, noninvasively, and in real-time measure CPE, eliminating the need for labels. In this study, two virus-permissive cell models were used to validate the impedance-TCID50 assay: the GFP-labeled adenovirus (Adv-GFP) in HEK293A cells and influenza A virus (IAV) in MDCK cells. The workflow is simple, encompassing cell seeding, virus inoculation, real-time monitoring, and automatic analysis of TCID50 by the software. Throughout infection, TCID50 values were automatically calculated by the system's software using the Reed-Muench formula at all recorded time points. TCID50 values of IAV obtained via impedance readouts were comparable to those generated by the conventional crystal violet staining-based TCID50 assay. These results demonstrate that virus quantification can be precisely and efficiently achieved using impedance measurement in combination with the Virology Module of the system's software. The new assay streamlines traditional methods while providing enhanced insight into viral dynamics, thereby supporting advanced virological research and expanding potential clinical applications.

Acute diffuse retinal hemorrhage is a rare but potentially serious complication following intravitreal anti-vascular endothelial growth factor (anti-vascular endothelial growth factor [VEGF]) therapy. Conbercept, a recombinant fusion protein, has been widely used for treating various retinal vascular disorders and has demonstrated both clinical efficacy and safety. Despite its widespread use, isolated reports of acute retinal hemorrhage post-injection have raised concerns. The case report presented here describes an episode of acute diffuse retinal hemorrhage occurring 24 h after the 12^th intravitreal conbercept (IVC) injection in a patient. The patient's significant risk factors included a long-standing history of poorly controlled diabetes mellitus, chronic hypertension, and advanced proliferative diabetic retinopathy (PDR). The patient had undergone pan-retinal photocoagulation in both eyes and received 11 bilateral IVC injections (0.5 mg/0.05 mL per eye) over a 12-month period. This report examines the underlying mechanisms, clinical course, and management, emphasizing the importance of personalized strategies for complex cases.

Recent advancements in large language models (LLMs) have led to notable improvements in abstractive summarization quality. However, hallucination - especially entity-level hallucination where non-existent or incorrect entities are introduced - remains a critical challenge. In this work, we propose a reward-driven fine-tuning framework for summarization models using the Entity Hallucination Index (EHI) as a guiding metric. The methodology here begins with generating initial summaries from pre-trained models such as Flan-T5, DistilBART, and Mistral (or other popular LLM) on structured transcript datasets, XSUM. We compute EHI by extracting named entities from both generated summaries and gold references, evaluating precision, and penalizing fabricated entities. The fine-tuning process is guided by reinforcement learning, where EHI serves as the reward signal. We adopt a REINFORCE-style update mechanism to optimize the summarization model towards maximizing entity faithfulness. Experiments demonstrate that models fine-tuned with EHI achieve lower hallucination rates without compromising informativeness. Furthermore, we show that EHI-guided models generalize better on out-of-domain summarization tasks, suggesting enhanced robustness. The approach here offers a practical direction for improving factuality in summarization, emphasizing the critical role of accurate entity representation.

Growing evidence underscores the importance of the first 1,000 days of life in shaping the gut-microbiome axis. This early-life window is critical in the establishment of long-term physiological trajectories and immunological adaptations, potentially influencing susceptibility to diseases linked to dysbiosis and barrier dysfunction. A deeper understanding of the underlying mechanisms requires a comprehensive assessment of key physiological parameters, including paracellular and transcellular permeability of the neonatal gut. Evaluating these parameters is essential to elucidate how early exposures to exogenous molecules may influence gut integrity and long-term health outcomes. Therefore, the first part of this article describes the oral administration of molecules of interest in mouse pups as early as day of life 6, while minimizing stress, risk of injury, and cannibalism. Lubricated, rounded-tip 24-G feeding needles are used to gavage mouse pups weighing a minimum of 2.5 g. The second part outlines the ex vivo assessment of paracellular and transcellular permeability using Ussing chamber assays on colonic samples from pups between DOL-10 and weaning. Pinless sliders adapted for small biopsies are used in combination with Ussing chambers to mount neonatal colonic samples. The paracellular probe FITC-Dextran 4 kDa and the transcellular marker horseradish peroxidase 44 kDa Type VI are added to the apical compartment of the system at the start of the assay. Samples are collected in the basolateral compartment at 0 min, 30 min, 60 min, 90 min, and 120 min to quantify probe passage. The passage of both markers is quantified directly (FITC) and indirectly (HRP) by a plate reader, calculated using standard curves, and expressed as flux.

Most existing studies on high-speed rail (HSR) seating comfort are based on subjective surveys and pressure maps; there is no physiological reporting on muscle fatigue and stress accrual with longer travel periods. The assessment of dynamic sitting performance in different travel conditions is rather difficult due to the impossibility of the traditional laboratory-based investigations to reproduce real-life posture variations. To surmount these limitations, the present research compares business, first-class, and economy-class seats in a real-world HSR travelling scenario through the application of multi-modal ergonomic testing, which involves the use of electromyography (EMG), skin conductance response (SCR), and scenario-based behavioral testing. The 30 participants were divided into two seating classes and placed in four functional situations: entertaining, dining, working, and resting. Whereas SCR was used to monitor autonomic stress responses, EMG was used to record muscular activity in the shoulders, lumbar, and neck. The results indicate that although business-class chairs reduce lumbar strain, they do not eliminate the weariness of upper limbs and necks, especially in the reclining position. The first-class seats are maximum in terms of working postures, but they cannot offer flexible support for varied body proportions, which results in head and lumbar discomfort. Owing to their low level of adjustability, economy-class seats produce the largest amount of muscular tension, despite their high level of lumbar and neck support. This research provides an understanding of adjustable, ergonomically idealized seating for trains in future models and the significance of incorporating behavioral and physiological information into dynamic seat analysis.

Accurate serotyping of Salmonella is essential for effective surveillance and outbreak investigation, as serotype diversity directly impacts pathogenicity and public health risk assessment. However, conventional slide agglutination methods are limited by poor reproducibility, labor intensity, and high costs, which hinder their application in high-throughput monitoring programs. To address these limitations, we developed and validated a genomic workflow integrating Multilocus Sequence Typing (MLST), the Salmonella In Silico Typing Resource (SISTR), SeqSero, SeqSero2, and SeqSero2S for serotype prediction using whole-genome sequencing data. This protocol was evaluated through a multicenter analysis of 315 Salmonella isolates collected from food and human sources in Southwest China. The findings of this study demonstrated significantly higher concordance among genomic approaches (up to 100%/99.1%/90.1% in the training set and 100%/97.1%/93.1% in the validation set for SISTR/MLST/SeqSero2S, respectively) compared to traditional serotyping. The workflow includes recommendations for selecting appropriate prediction methods based on surveillance context, emphasizing MLST and SeqSero2S for routine monitoring, SeqSero2S for rapid screening, and SISTR with core genome MLST for outbreak investigations. This approach facilitates the integration of genomic serotyping into public health practice, reducing reliance on traditional serology and improving reproducibility and scalability in Salmonella monitoring programs.

Bile duct injuries during cholecystectomy are most frequently attributed to misidentification of surgical anatomy. Initial management following a major bile duct injury is time-sensitive and critical to the patient's clinical course and overall outcome. We present a 63-year-old male patient who sustained a common bile duct transection during laparoscopic cholecystectomy at a resource-limited rural surgery center. Upon recognition of the injury, an intraoperative video consultation was made to hepatobiliary surgery at our academic referral institution, providing a real-time explanation of the dissection with direct visualization of surgical anatomy. This expedited sequence facilitated acquisition of diagnostic studies, including an intraoperative cholangiogram, confirming common bile duct transection, and a CT angiogram, demonstrating intact arterial anatomy. Direct to OR transfer to the tertiary facility was therefore prioritized, circumventing a prolonged bed wait the patient would have otherwise required while awaiting injury diagnosis and characterization. He underwent definitive repair with robotic-assisted Roux-en-Y hepaticojejunostomy on the same day that the injury occurred. This paper provides an operative video with a concise and adoptable method for performing a definitive hepaticojejunostomy following bile duct transection intended for fellowship-trained hepatobiliary surgeons.

The integration of gamification in mobile-assisted language learning (MALL) has reshaped how learners can engage with vocabulary acquisition and self-directed communication. Few studies have examined the impact of gamified applications on learner behavior and psychological reactions, particularly in non-native English learning contexts, despite their frequent commendation for enhancing user motivation and engagement. This study examines the motivational impact and ideological underpinnings of popular gamified English vocabulary apps, using Fairclough's three-dimensional Critical Discourse Analysis (CDA) framework to analyze both user interactions and platform design. Drawing on interface data, user sentiment analysis, and interview transcripts from the Chinese university students studying English as a foreign language (EFL), this study reveals that, consistent with Self-Determination Theory (SDT), gamification plays a significant role in encouraging learner motivation and engagement but simultaneously embeds neoliberal ideals of competition, self-optimization, and behavioral monitoring. These features, while intended to boost performance, may undermine intrinsic motivation over time. The findings highlight the psychological and sociotechnical dynamics of gamified learning environments, offering new insight into how digital educational platforms can reinforce self-regulatory expectations and performance-oriented discourse.

Colostomy substantially affects daily function and psychosocial status, and middle-aged and elderly patients often experience reduced self-care ability, heightened anxiety/depression, and early complications. This single-center retrospective cohort evaluated whether a standardized, professional nursing pathway improves short-term outcomes after colostomy compared with routine care. We analyzed 80 consecutive patients who underwent colostomy between January 2022 and December 2023 and received either routine care (n = 40) or a multi-module pathway (n = 40) integrating structured education, standardized stoma and peristomal skin care, individualized lifestyle/diet guidance, nurse-led brief cognitive-behavioral support, and scheduled follow-up. Outcomes at baseline and post 1 month included self-care ability (Self-Care Ability Assessment Scale, ESCA), anxiety (Self-Rating Anxiety Scale, SAS), depression (Self-Rating Depression Scale, SDS), quality of life (City of Hope Quality of Life-Ostomy Questionnaire, COH-QOL-OQ), complication rates (e.g., stoma infection, stenosis), and nursing satisfaction assessment. Compared with routine care, the professional-care group showed significantly greater improvements in ESCA and COH-QOL-OQ and larger reductions in SAS/SDS (all p < 0.05), along with lower early complication rates and higher nursing satisfaction. This real-world data suggests that a standardized, nurse-delivered post-colostomy pathway improves short-term clinical and patient-reported outcomes in middle-aged and elderly patients.