Journal Of Visualized Experiments[JOURNAL]
Sun
200 papers
Thyroid Artery Flow Velocity and Serum Adenosine Deaminase After Methimazole Therapy in Autoimmune Hyperthyroidism.
He Y, Huang D, Yang T
J Vis Exp · 2026 Jun · PMID 42329883 · Publisher ↗
Autoimmune hyperthyroidism, including Graves' disease and Hashimoto's hyperthyroidism, presents with overlapping biochemical features but distinct pathophysiological mechanisms. Non-invasive methods that reflect both vas...
Autoimmune hyperthyroidism, including Graves' disease and Hashimoto's hyperthyroidism, presents with overlapping biochemical features but distinct pathophysiological mechanisms. Non-invasive methods that reflect both vascular and immune activity may improve disease differentiation and treatment monitoring. This retrospective single-center study included 60 newly diagnosed hyperthyroid patients (30 Graves' disease and 30 Hashimoto's hyperthyroidism). All participants underwent baseline and 12-week follow-up evaluations after methimazole therapy. Assessments included thyroid function tests, superior thyroid artery Doppler parameters (peak systolic velocity [PSV], resistance index [RI]), and serum adenosine deaminase (ADA) levels. Statistical analyses included paired and independent t-tests and Pearson correlation. At baseline, PSV and ADA levels were higher in Graves' disease compared to Hashimoto's hyperthyroidism. After 12 weeks of methimazole treatment, both groups showed significant reductions in PSV and ADA (p < 0.05), with greater changes observed in Graves' disease. Moderate correlations were observed between ADA levels and Doppler parameters. The combined assessment of Doppler ultrasonography and serum ADA provides a reproducible, non-invasive approach to evaluate vascular and immune changes during therapy. While the method shows potential for differentiating autoimmune hyperthyroid subtypes and monitoring treatment response, further validation in larger cohorts is required.
A Semi-automated Method For Detecting Group Differences In Protocerebral Anterior Medial Cluster Dopaminergic Neuron Numbers In The Drosophila Brain.
Ren M, Lim KL
J Vis Exp · 2026 Jun · PMID 42329882 · Publisher ↗
Drosophila melanogaster serves as a powerful model organism for studying neurodegenerative diseases, including Parkinson's disease (PD), primarily through the analysis of dopaminergic (DA) neuron degeneration and associa...
Drosophila melanogaster serves as a powerful model organism for studying neurodegenerative diseases, including Parkinson's disease (PD), primarily through the analysis of dopaminergic (DA) neuron degeneration and associated locomotion deficits. Historically, quantification of DA neuron loss in the fly brain has been confined to smaller, readily countable clusters, including protocerebral posterior medial (PPM) clusters and protocerebral posterior lateral (PPL) clusters. In contrast, the substantially larger protocerebral anterior medial (PAM) cluster, comprising over 100 neurons, presents a significant challenge for manual counting, leading to its underrepresentation in quantitative studies. To date, reported investigations still rely on labor-intensive manual counts through confocal Z-stacks, which are time-consuming and prone to variability. To address this methodological gap, we developed a semi-automated image analysis pipeline using the open-source tool Labkit. This protocol enables efficient group-relative quantification of tyrosine hydroxylase (TH)-positive neurons in the PAM cluster within the same experimental batch. The method starts with standard immunofluorescence staining and confocal imaging, followed by a detailed, step-by-step guide for segmentation and analysis. This approach addresses the bottleneck of PAM cluster quantification and provides a practical workflow for detecting intergroup differences in DA neuron vulnerability in models of neurodegeneration. The relative scarcity of PD studies on the PAM cluster reflects a historical technical barrier rather than a lack of biological relevance. By overcoming this barrier, our method opens the door to systematic investigation of PAM neuron vulnerability in PD and its potential link to non-motor symptoms.
Effect of Bone Cement Viscosity on Leakage and Outcomes in Vertebroplasty.
Chen X, Yuan Z
J Vis Exp · 2026 Jun · PMID 42329875 · Publisher ↗
Percutaneous vertebroplasty provides rapid pain relief in elderly patients with acute osteoporotic vertebral compression fractures; however, bone cement leakage remains a major safety concern, and the influence of cement...
Percutaneous vertebroplasty provides rapid pain relief in elderly patients with acute osteoporotic vertebral compression fractures; however, bone cement leakage remains a major safety concern, and the influence of cement viscosity on leakage and clinical outcomes is not fully established. This study evaluated whether high-viscosity cement reduces leakage without compromising efficacy or safety. In this single-center, prospective randomized controlled trial, 184 elderly patients with single-level acute osteoporotic vertebral compression fractures were enrolled and followed for 12 months. Traumatic burst fractures and cases with posterior wall cortical disruption causing >25% spinal canal compromise were excluded. Participants were randomized 1:1 to receive high- or low-viscosity bone cement. The primary outcome was overall cement leakage at 48 h postoperatively, assessed by computed tomography and classified using the Yeom system. Secondary outcomes included pain (Visual Analog Scale), function (Oswestry Disability Index), radiographic parameters, perioperative variables, and adverse events. High-viscosity cement significantly reduced the overall leakage rate compared with low-viscosity cement (RR 0.47, 95% CI 0.29-0.78; adjusted OR 0.36, 95% CI 0.19-0.71; P = 0.003), with consistent results in sensitivity analyses. Leakage patterns differed between groups, with C-type leakage more frequent in the high-viscosity group and B-type leakage more frequent in the low-viscosity group (P = 0.008). Pain and functional outcomes improved significantly in both groups, with no between-group differences over time (all P > 0.05). Radiographic correction, perioperative parameters, and adverse event rates were comparable. Image interpretation reliability was high (κ = 0.86; ICC > 0.90). High-viscosity bone cement reduces early leakage while maintaining comparable clinical and radiographic outcomes, supporting its use as a preferred option in vertebroplasty for this population.
MiR-483-3p as a Prognostic Marker In Non-Small Cell Lung Cancer: Suppression of Tumor Progression via KIF3B Downregulation.
Yang J, Wang W
J Vis Exp · 2026 Jun · PMID 42329872 · Publisher ↗
MicroRNAs play wide roles in non-small cell lung cancer (NSCLC). To investigate the clinical value of miR-483-3p in NSCLC and its molecular target, 350 NSCLC patients were recruited in this study. RT-qPCR results showed...
MicroRNAs play wide roles in non-small cell lung cancer (NSCLC). To investigate the clinical value of miR-483-3p in NSCLC and its molecular target, 350 NSCLC patients were recruited in this study. RT-qPCR results showed that tumor histological expression of miR-483-3p progressively decreased as the tumor-node metastasis (TNM) stage increased. The receiver operating characteristic (ROC) curve displayed that histological miR-483-3p level can effectively distinguish NSCLC patients with high TNM stage (III) from those with low stage (I+II) (area under the ROC curve = 0.869). The Kaplan-Meier curve showed that NSCLC patients with low miR-483-3p expression demonstrated a lower 5-year overall survival rate. After adjusting for other confounding factors, multivariate Cox analysis further identified miR-483-3p as an independent protective factor for NSCLC survival. Mechanistically, RNA pull-down assay showed that upregulation of miR-483-3p was co-precipitated with KIF3B mRNA and inhibited its expression, thereby suppressing the malignant phenotypes of NSCLC cells and inducing apoptosis. In conclusion, downregulation of miR-483-3p serves as a poor prognostic marker in NSCLC, potentially affecting cancer progression by negatively regulating KIF3B.
Targeted Sedation And Circadian Interventions In Mechanically Ventilated ICU Patients: A Narrative Review.
Li J, Su R, Lin Y … +5 more , Wang Y, Wu Y, Deng XD, Li B, Iv S
J Vis Exp · 2026 Jun · PMID 42329869 · Publisher ↗
Circadian rhythms regulate sleep-wake cycles, hormone secretion, immune functions, and cardiovascular activity. However, they are frequently disrupted in mechanically ventilated ICU patients due to critical illness, envi...
Circadian rhythms regulate sleep-wake cycles, hormone secretion, immune functions, and cardiovascular activity. However, they are frequently disrupted in mechanically ventilated ICU patients due to critical illness, environmental factors, and sedative exposure. This narrative review aims to examine the interaction between sedation practices, environmental interventions, and circadian disruption. A structured, non-systematic search of MEDLINE/PubMed, Web of Science, and Scopus (200-2025) was performed to identify mechanistic and clinical studies. Evidence indicates that deep and prolonged sedation is associated with decreased melatonin secretion, disrupted sleep architecture, and increased delirium risk, whereas minimal sedation combined with circadian-aligned non-pharmacological interventions, such as light and noise modulation and care clustering, may support organization. However, findings remain conflicting and are largely derived from observational studies. The use of a combined sedation-circadian rhythms care strategy represents a novel, untested approach. Future studies should incorporate standardized circadian outcomes and controlled designs to establish causality and evaluate clinical efficacy.
Transcriptome Sequencing Identified Hub Pathogenic Chemokines In Atherosclerotic Plaques.
Li Z, Wang P, Si J … +7 more , Fan J, Guo H, Lu G, Wang Q, Wang J, Hao J, Zhang L
J Vis Exp · 2026 Jun · PMID 42329864 · Publisher ↗
Atherosclerotic plaque (AP) is a kind of inflammatory fibrous tissue proliferative disease after the injury of endothelial cells and smooth muscle cells in the artery wall, which can lead to different degrees of cardiova...
Atherosclerotic plaque (AP) is a kind of inflammatory fibrous tissue proliferative disease after the injury of endothelial cells and smooth muscle cells in the artery wall, which can lead to different degrees of cardiovascular and cerebrovascular blood flow obstruction. However, effective therapeutic approaches targeting inflammation have largely failed to date, suggesting that additional insights remain needed. In this study aimed to analyze inflammatory factor-related transcriptomic changes in AP using RNA-sequencing (RNA-seq). RNA-seq was performed on samples from AP patients (n = 11) and control individuals (n = 3). Differentially expressed genes (DEGs) were identified using Metascape, followed by KEGG pathway enrichment analysis using clusterProfiler package in R software, immune infiltration analysis using CIBERSORT, and protein-protein interaction (PPI) network using STRING database. Hub genes within the PPI network were identified using the CytoHubba plugin. A total of 3713 DEGs were identified in AP group, including 2097 up-regulated and 1616 down-regulated genes. The results showed that DEGs were mainly enriched in immune- and inflammation-related pathways. Three inflammation-related factors CCL3, CCL4, and CXCL1 were considered as major hub genes in the pathological process of AP. Immune infiltration analysis revealed a distinct microenvironment in AP, characterized by a significant increase in M0 macrophages alongside reductions in CD8⁺ T cells, activated NK cells, and resting mast cells within AP. In conclusion, these procedural findings describe the differenrial expression of CCL3, CCL4, and CXCL1 and their association with an altered immune cell composition in AP, highlighting the three chemokines as potential candidates for further mechanistic investigation.
A Reproducible Survey-Scoring And C5.0 Decision-Tree Workflow For Classifying Self-Reported Higher-Order Thinking In Generative AI-Supported Learning.
Zhao X, Song P, Zhong M … +1 more , Zhu S
J Vis Exp · 2026 Jun · PMID 42329863 · Publisher ↗
Generative artificial intelligence is increasingly used in higher education, but researchers still need transparent procedures for collecting, scoring, and organizing learner-level survey data in this rapidly changing co...
Generative artificial intelligence is increasingly used in higher education, but researchers still need transparent procedures for collecting, scoring, and organizing learner-level survey data in this rapidly changing context. This article presents a reproducible survey-scoring and C5.0 decision-tree workflow for classifying self-reported higher-order thinking among college students who have used generative artificial intelligence tools for academic learning. The protocol covers participant recruitment, questionnaire administration, response-quality screening, composite-score calculation, binary coding, training-testing partitioning, C5.0 tree construction, pruning, model-output export, and interpretation of node-based classification paths. The workflow is demonstrated using a single-university survey dataset of 776 undergraduate students collected in China from March 7 to March 15, 2023. Higher-order thinking is operationalized as a self-reported questionnaire score rather than as directly observed cognitive performance. In the demonstration dataset, the pruned tree retained eight learner-related variables: generative artificial intelligence anxiety, trust in generative artificial intelligence, problematic smartphone use, academic procrastination, academic performance, parental upbringing, negative emotions, and attitudes toward generative artificial intelligence. The retained model achieved 89.52% accuracy in the training subset and 86.21% accuracy in the testing subset. However, class-specific evaluation showed uneven performance, with substantially weaker recall for the Low-HOT class than for the High-HOT class. Therefore, the tree should be interpreted as an auxiliary, interpretable workflow demonstration rather than as a validated screening tool. This protocol may support researchers who require a documented and repeatable procedure for survey-based learner profiling in generative artificial intelligence-supported learning environments.
Heart Disease Prediction Using Statistical Feature Selection and Interpretable Machine Learning.
Babers R, Madbouly AMM
J Vis Exp · 2026 Jun · PMID 42329851 · Publisher ↗
Heart disease is a major cause of death worldwide, making its early prediction an important clinical and computational issue. Several studies have addressed challenges such as data scarcity, feature selection, and model...
Heart disease is a major cause of death worldwide, making its early prediction an important clinical and computational issue. Several studies have addressed challenges such as data scarcity, feature selection, and model interpretability individually, but fewer studies have proposed an integrated framework that addresses these challenges in a synergistic manner. This paper presents a comprehensive predictive framework that uses: (1) a generative adversarial network (GAN) to address class imbalance and data scarcity; (2) a hybrid feature selection approach that combines statistical pre-filtering via Welch's t.-test and Cohen's d effect size, along with metaheuristic optimization via Harris Hawk Optimization; and (3) various explainable artificial intelligence methods, including SHAP, partial dependence plots, and odds ratios. This framework was assessed on the Cleveland and Statlog datasets, yielding strong accuracy, F1-scores, and ROC-AUC values compared with selected baselines and existing methods. The model provides a robust, interpretable computational framework for heart disease prediction, linking machine learning performance with clinical interpretability.
Comparative Evaluation of Deep Learning Model Complexity for Forecasting Non-Ferrous Metal Prices.
Li L, Guan T, Li L … +1 more , Zeng J
J Vis Exp · 2026 Jun · PMID 42329850 · Publisher ↗
This study examines whether increasing architectural complexity improves forecasting accuracy in deep learning-based financial models. Using daily spot price data from the Shanghai Metals Market for copper (Cu), aluminum...
This study examines whether increasing architectural complexity improves forecasting accuracy in deep learning-based financial models. Using daily spot price data from the Shanghai Metals Market for copper (Cu), aluminum (Al), and zinc (Zn) spanning January 2015 to September 2025, a standardized preprocessing pipeline was applied, including z-score normalization and sliding window sequence construction (window length = 30, forecast horizon = 1). A total of eighteen models were systematically evaluated, including gated recurrent units (GRUs), long short-term memory (LSTM) networks, convolutional neural network-bidirectional LSTM-attention hybrids (CNN-BiLSTM-Attention), as well as traditional econometric models (autoregressive integrated moving average and generalized autoregressive conditional heteroskedasticity), machine learning models (random forest and extreme gradient boosting), and a Transformer-based model. All deep learning models were trained exclusively on Cu data and evaluated on independent Al and Zn datasets to assess generalizability. Results show that the standard GRU model achieves the lowest error rates (mean absolute error [MAE] = 1032.85; root mean square error = 1344.30) and highest explanatory power (coefficient of determination [R] = 0.907) on the Cu test set, while also performing strongly on Al (MAE = 167.51, R = 0.918) and Zn (MAE = 254.23, R = 0.952). Ablation analysis demonstrates that adding architectural components such as attention mechanisms, bidirectional layers, and convolutional modules reduces predictive accuracy. Statistical testing using the Diebold-Mariano test indicates that most performance differences are significant (p < 0.05). These findings highlight the limitations of unnecessary model complexity and support the use of simpler, robust approaches for commodity price forecasting.
Age-stratified Evaluation Of Saccharomyces boulardii In A Pediatric Randomized Controlled Trial And Adult Observational Study of Infectious Enteritis.
Lou Y, Wang Q, Xing X
J Vis Exp · 2026 Jun · PMID 42329849 · Publisher ↗
Infectious enteritis remains a significant global health burden, particularly among children under five years of age. Saccharomyces boulardii (S. boulardii), a probiotic yeast, has been widely studied for its role in red...
Infectious enteritis remains a significant global health burden, particularly among children under five years of age. Saccharomyces boulardii (S. boulardii), a probiotic yeast, has been widely studied for its role in reducing diarrhea; however, direct comparisons of its effects across age groups remain limited. This study aims to evaluate and compare the efficacy and safety of S. boulardii in pediatric and adult patients with infectious enteritis. A double-blind, randomized, placebo-controlled trial was conducted in 186 children with acute diarrhea, while a retrospective observational study included 270 adult patients. All participants received standard rehydration therapy; the intervention groups additionally received S. boulardii. The primary outcome was diarrhea duration. Secondary outcomes included stool frequency, length of hospital stay, recurrence rate, and adverse events. In pediatric patients, S. boulardii significantly reduced diarrhea duration (2.1 vs. 3.9 days in rotavirus-positive cases), hospital stay (3.2 vs. 4.8 days), and stool frequency (4.2 vs. 6.7 stools/day) compared to placebo (all p < 0.05). In adult patients, S. boulardii was associated with shorter diarrhea duration (2.7 vs. 5.4 days), reduced hospital stay (3.6 vs. 5.2 days), higher symptom resolution rates (77.8% vs. 15.8%), and lower recurrence (8.1% vs. 21.7%; all p < 0.01). Adverse events were mild and comparable between groups. S. boulardii appears to be a safe and effective adjunct therapy for infectious enteritis, with age-related differences in clinical outcomes. S. boulardii appears to be a safe and effective adjunct therapy for infectious enteritis in both pediatric and adult populations. Within each cohort, improvements were observed in clinical outcomes, including diarrhea duration, hospitalization, and recurrence. Observed differences between pediatric and adult cohorts are descriptive and reflect separate analyses conducted under different study designs; therefore, no direct statistical comparisons between age groups were performed.
Long-Term Outcomes and Predictors of Response to Anti-VEGF Therapy for Diabetic Macular Edema in Clinical Practice.
Dong L, Tian J
J Vis Exp · 2026 Jun · PMID 42329847 · Publisher ↗
This protocol provides a standardized methodology for evaluating the 24-month effectiveness and safety of anti-vascular endothelial growth factor (anti-VEGF) therapy in center-involved diabetic macular edema (DME) using...
This protocol provides a standardized methodology for evaluating the 24-month effectiveness and safety of anti-vascular endothelial growth factor (anti-VEGF) therapy in center-involved diabetic macular edema (DME) using real-world clinical data. The procedure involves a structured electronic medical record (EMR) screening algorithm based on ICD-10 codes, followed by manual clinical verification. Patients are categorized into a class-level anti-VEGF treatment group (receiving ranibizumab, aflibercept, or conbercept) or a non-biologic control group (focal/grid laser or observation). To mitigate non-random allocation bias inherent in retrospective data, inverse probability of treatment weighting (IPTW) is employed to balance baseline demographic and tomographic covariates across groups. The methodology specifies standardized visual acuity assessment using ETDRS charts and detailed OCT acquisition parameters to ensure longitudinal data consistency. By analyzing visual and anatomical trajectories alongside structural biomarkers, such as the disorganization of the retinal inner layers (DRIL), this approach provides a reproducible framework for assessing long-term outcomes in routine clinical settings. While acknowledging the limitations of a single-center retrospective design, these methods offer critical evidence to help characterize the efficacy-effectiveness gap and optimize personalized management strategies for DME.
No Significant Direct Causal Association Between TNF Pathway Biomarkers and Hip Fracture Risk: A Study Based on Real-World Data.
Xiao P, Wei B, Zhang G … +11 more , Liu D, Zhang Y, Sun J, Wan H, Heng Y, Zhou S, Li Y, Ren X, Sun Z, Huang Y, Fu X
J Vis Exp · 2026 Jun · PMID 42329845 · Publisher ↗
Hip fracture is a common and serious condition in elderly individuals, often associated with altered TNF-α signaling. However, few studies have explored whether changes in TNF pathway biomarkers contribute to the risk of...
Hip fracture is a common and serious condition in elderly individuals, often associated with altered TNF-α signaling. However, few studies have explored whether changes in TNF pathway biomarkers contribute to the risk of hip fracture. In this study, adverse events related to hip fracture reported for five TNF inhibitors were analyzed using data from the first quarter of 2014 to the fourth quarter of 2023. After data standardization and cleaning, four disproportionality methods, including the reporting odds ratio, proportional reporting ratio, multi-item gamma Poisson shrinker, and Bayesian confidence propagation neural network, were applied to assess the association between TNF inhibitor exposure and hip fracture outcomes. Complementary mendelian randomization analyses were further conducted using TNF-α, sTNFR1, and sTNFR2 as exposures and hip fracture as the outcome. Pharmacovigilance analyses revealed no significant association between TNF inhibitor exposure and hip fracture-related adverse events across the four algorithms. Mendelian randomization analyses identified no significant direct causal association between genetically predicted TNF-α and hip fracture risk, and additional analyses of sTNFR1 and sTNFR2 yielded consistent results. These findings suggest that TNF pathway biomarkers are unlikely to act as independent direct determinants of hip fracture risk and may provide a useful basis for future mechanistic studies and prevention strategies.
A Propensity Score-Matched Protocol: Terazosin Plus Finasteride For Benign Prostatic Hyperplasia - Sex Hormone And Urinary Control Effects.
Wu Y, Ma W
J Vis Exp · 2026 Jun · PMID 42329844 · Publisher ↗
To investigate the clinical effect of terazosin hydrochloride combined with finasteride in benign prostatic hyperplasia (BPH), and to analyze its effect on sex hormones and urinary control. This retrospective propensity...
To investigate the clinical effect of terazosin hydrochloride combined with finasteride in benign prostatic hyperplasia (BPH), and to analyze its effect on sex hormones and urinary control. This retrospective propensity score‑matched (PSM) cohort study included BPH patients treated between January 2020 and March 2021. 1:1 PSM using logistic regression with prespecified covariates (age, BMI, IPSS, Qmax, PVR, PV, PSA, CRP, T, E2, DHT, incontinence episodes, ICIQ‑SF); inclusion criteria (age 50‑80 years, IPSS ≥ 8, PSA > 1.4 ng/mL, PV ≥ 40 mL, Qmax ≤ 15 mL/s); exclusion of prior prostate surgery or 5α‑reductase/α1‑blocker use; 12‑month treatment and 36 month follow up with outcomes measured at 6, 12, 24, 36 months. Sixty patients were allocated to each group after PSM. The combination group showed significantly greater improvements in IPSS, Qmax, PVR, and PV (all P < 0.05), lower DHT and E2, higher T (all P < 0.05), fewer 24 h incontinence episodes, and better ICIQ‑SF scores (both P < 0.05). Safety profiles were comparable (P > 0.05). The terazosin‑- finasteride regimen is associated with improved long-term outcomes, better urinary control, and favorable hormonal changes, though hormonal effects are primarily attributable to finasteride.
From Tissue to Morphometry: A Step-by-Step Golgi-Cox Workflow for Neuronal and Dendritic Spine Analysis in Adult Mice.
Rubio-de Anda PF, Martínez-Orozco H, Díaz Miranda SY
J Vis Exp · 2026 Jun · PMID 42329838 · Publisher ↗
Neuronal morphology analysis in health and disease is fundamental for understanding brain function and for assessing the effects of experimental interventions. Golgi-Cox staining remains a classical, yet highly relevant...
Neuronal morphology analysis in health and disease is fundamental for understanding brain function and for assessing the effects of experimental interventions. Golgi-Cox staining remains a classical, yet highly relevant technique for visualizing complete neuronal arbors and dendritic spines. However, published protocols frequently differ in critical procedural steps and often lack essential methodological details, leading to substantial variability in staining quality and making laboratory standardization time-consuming and challenging. Here, a comprehensive, step-by-step Golgi-Cox protocol for adult mice is presented, encompassing the entire workflow from in vivo handling and tissue processing to imaging and morphometric data extraction. It details practical considerations and common pitfalls that influence impregnation efficiency, section integrity, and signal-to-noise ratio, and provide methodological guidance to improve reproducibility. Overall, this protocol enables reliable visualization and quantitative analysis of whole neurons, dendritic segments, and dendritic spines. It is anticipated that this resource will facilitate the implementation of Golgi-Cox staining with greater consistency, transparency, and technical rigor, while offering a clear framework for adapting the method to diverse laboratory settings.
A Reproducible Survey Workflow for Examining Psychological Resilience, Sense of School Belonging, and Higher-Order Thinking in College Students.
Zhong M, Li C, Song P … +2 more , Liu X, Yang P
J Vis Exp · 2026 Jun · PMID 42329835 · Publisher ↗
This method article presents a reproducible cross-sectional self-report survey protocol for examining associations among psychological resilience, sense of school belonging (SSB), higher-order thinking, and internet gami...
This method article presents a reproducible cross-sectional self-report survey protocol for examining associations among psychological resilience, sense of school belonging (SSB), higher-order thinking, and internet gaming disorder (IGD) in college students. In this protocol, new employment patterns are treated as a contextual background shaped by generative artificial intelligence, platform-based work, and changing graduate skill expectations rather than as a participant-level exposure variable. The protocol integrates institutional sampling, questionnaire administration, translation documentation, response-quality screening, missing-data handling, scale scoring, common-method-bias screening, variable centering, and conditional process analysis into one standardized workflow. SSB is specified as a statistical mediator, and IGD is specified as a moderator within an association-based moderated mediation framework. Representative outputs include sample-flow records, descriptive statistics, correlation matrices, mediation and moderated mediation estimates, conditional indirect effects, the index of moderated mediation, and interaction plots. Because the representative dataset is cross-sectional and self-reported, the protocol supports transparent estimation of statistical associations rather than causal or longitudinal inference. By standardizing preprocessing decisions and model specifications before analysis, this workflow improves transparency, comparability, and reproducibility in educational and behavioral survey research.
Haemoperfusion Combined With Xuebijing Injection Improves Inflammatory Status And Prognosis In Patients With End-Stage Renal Disease.
Liang F, Yang Y, Wang M … +4 more , Zhang F, Xing L, Sun J, Gao J
J Vis Exp · 2026 Jun · PMID 42329834 · Publisher ↗
End-stage renal disease (ESRD) is an irreversible, chronic, progressive disease that seriously affects patients' quality of life, and clinical treatment currently faces certain challenges. A total of 100 ESRD patients fr...
End-stage renal disease (ESRD) is an irreversible, chronic, progressive disease that seriously affects patients' quality of life, and clinical treatment currently faces certain challenges. A total of 100 ESRD patients from the Affiliated Hospital of North China University of Science and Technology were enrolled in this retrospective study and divided into HT and HI groups based on treatment modality. Both groups were treated with haemoperfusion, and the HI group was additionally treated with Xuebijing injection. The main assessments of the two groups were inflammatory factor indexes, immune indexes, renal function indexes, and clinical efficacy. Secondary outcomes included quality of life [Health Questionnaire (SF-36) score], complication and adverse reaction rates. Patient survival rates were calculated to analyze the correlation between microinflammatory status and prognosis. Logistic regression and receiver operating characteristic (ROC) analyses were conducted to further explore prognostic factors and predict complication risk. After treatment, the indicators in both groups differed significantly compared with the pre-treatment values (P < 0.05). Inflammatory indexes, renal function indexes, complication rates, and adverse reaction rates in the HI group were lower than those in the HT group, whereas immune indexes, clinical efficacy, SF-36 score, and survival rate were higher than those in the HT group (P < 0.05). Logistic regression analysis and ROC results showed that inflammatory indexes, immune indexes, and renal function indexes were closely associated with prognosis (P < 0.05). Combined therapy of haemoperfusion and Xuebijing shows favorable clinical efficacy in ESRD patients, improving renal function, alleviating the inflammatory response, regulating immune function, and contributing to favorable prognostic outcomes, and warrants further clinical exploration and validation.
Simulating High-Altitude Hypoxic Conditions Delays Wound Healing In Rats.
Cai S, Yao G, Lin L … +9 more , Chen Z, Linbo J, Zhou Z, Zhou S, Wang P, Cai Z, Fan D, Lin Y, Zhang Y
J Vis Exp · 2026 Jun · PMID 42329831 · Publisher ↗
High-altitude hypobaric conditions may cause prolonged wound hypoxia and impaired healing, yet standardized models remain scarce. In this study, we developed a wound delayed healing model and preliminarily validated it b...
High-altitude hypobaric conditions may cause prolonged wound hypoxia and impaired healing, yet standardized models remain scarce. In this study, we developed a wound delayed healing model and preliminarily validated it by simulating high-altitude hypoxia using a controlled hypobaric system. Rats were housed at simulated altitudes of 3000 m (air pressure: 70.1 kPa, partial oxygen pressure: 14.7 kPa) to 8000 m (air pressure: 35.6 kPa, partial oxygen pressure: 7.5 kPa) to determine safety thresholds, with oxidative stress and skin hypoxia assessed. Results showed increased mortality risk at simulated 6000 m altitude, with tolerance observed below 5000 m. Elevated altitudes were associated with worsening oxidative stress and increased skin hypoxia; exhibited altitude-dependent delays in wound healing, reduced perfusion, suppressed collagen remodeling, and exacerbated inflammatory responses. Despite upregulation of vascular regenerative factors, microvascular density decreased. Overall, this study developed and preliminarily validated a rat model suitable for investigating mechanisms and interventions in high-altitude hypoxia-induced chronic wounds.
Aconitine Induces Myocardial Damage in Rats Associated With Activation of the JNK1/2 Signaling Pathway.
Hu C, Huang Y, Hu W … +2 more , Yang R, Wu H
J Vis Exp · 2026 Jun · PMID 42329830 · Publisher ↗
Acute Aconitine (Acon) poisoning can cause arrhythmia and myocardial damage; however, the underlying mechanisms remain unclear. This study investigated the mechanism of myocardial injury induced by acute Acon poisoning....
Acute Aconitine (Acon) poisoning can cause arrhythmia and myocardial damage; however, the underlying mechanisms remain unclear. This study investigated the mechanism of myocardial injury induced by acute Acon poisoning. A rat model (n = 51) of acute Acon poisoning was established by intragastric administration. Electrocardiographic tracings were used to record changes in cardiac rhythm. Hematoxylin and eosin (HE) staining and Masson staining were performed to evaluate myocardial pathological injury and collagen deposition. The expression levels of cardiac enzymes and proteins in venous blood were detected using enzyme-linked immunosorbent assay (ELISA) kits. The structure-activity relationship between Acon and the JNK pathway was investigated by molecular docking. The expression levels of phosphorylated c-Jun N-terminal kinase 1/2 (JNK1/2) proteins were detected by Western blotting. In this study, ventricular arrhythmias were observed in rats following intragastric administration of Acon. Myocardial tissue showed a disordered arrangement of myocardial fibers and cells, accompanied by abnormal collagen deposition. Molecular docking results indicated potential interactions between Acon and JNK1/2. Meanwhile, the expression levels of cardiac enzymes and proteins, as well as phosphorylated JNK1/2, were significantly increased. Further studies demonstrated that treatment with the JNK inhibitor SP600125 partially reversed Acon-induced myocardial injury in rats. These results indicate that JNK1/2 phosphorylation is involved in the development of myocardial injury induced by acute Aconitine poisoning. The findings suggest that activation of the JNK1/2 signaling pathway contributes to Aconitine-induced myocardial damage, and that inhibition of the JNK1/2 pathway may alleviate myocardial injury caused by acute Aconitine intoxication.
Quantitative Live Cell Imaging of Nuclear Shape and Chromatin Dynamics During Development and Environmental Stress in Arabidopsis thaliana Root.
Demura-Devore J, Ashraf MA
J Vis Exp · 2026 Jun · PMID 42329829 · Publisher ↗
The nucleus is the characteristic organelle of eukaryotic organisms. Unlike the classic textbook view of static nuclei, nuclear shape is dynamic in live cells. Altered or deformed nuclear shape is a hallmark of cancer in...
The nucleus is the characteristic organelle of eukaryotic organisms. Unlike the classic textbook view of static nuclei, nuclear shape is dynamic in live cells. Altered or deformed nuclear shape is a hallmark of cancer in animal cells and environmental stress in plants. Nuclear envelope proteins interact with chromatin to regulate gene expression. Unfortunately, little is known about the impact of abiotic stress on nuclear shape, movement, and chromatin dynamics. To confront this issue, we developed a pipeline using confocal microscopy and particle tracking software to quantify nuclear and chromatin dynamics in Arabidopsis roots under control and abiotic stress condition. This confocal imaging method utilizes a dual fluorescently tagged marker line - nuclear envelope protein and chromatin - to perform live cell imaging of the root in model plant Arabidopsis thaliana under control and salt-stressed conditions. These captured movies are analyzed to quantify nuclear and chromatin dynamics using open-source image processing software Fiji/ImageJ with the help of the TrackMate plugin. To validate this method, we imaged and quantified chromatin movement in control and salt-stressed roots, revealing a decrease in chromatin speed under salt-stressed conditions. This method allows for quantitative live cell imaging of root nuclear shape and chromatin dynamics during plant development and environmental stress, thus enabling analysis of changes in nuclear and chromatin dynamics caused by abiotic stressors.
Age-Specific Pyroptosis Biomarker Analysis and Non-Pharmacological Intervention in Acute Respiratory Distress Syndrome.
Chen Y, Liu T, Zeng Y … +7 more , Chen Y, Zou Q, Kong Y, Li Y, Shen G, Wang X, Li Y
J Vis Exp · 2026 Jun · PMID 42329827 · Publisher ↗
Acute respiratory distress syndrome (ARDS) is a life-threatening inflammatory lung disorder with high morbidity and mortality, in which pyroptosis has emerged as a pivotal pathogenic mechanism. Considering that age-relat...
Acute respiratory distress syndrome (ARDS) is a life-threatening inflammatory lung disorder with high morbidity and mortality, in which pyroptosis has emerged as a pivotal pathogenic mechanism. Considering that age-related immune and molecular differences may influence pyroptosis and therapeutic response, this study aimed to identify age-specific pyroptosis-associated biomarkers and evaluate the therapeutic potential of spectrum energy water (SEW) combined with far-infrared radiation (FIR). The protocol's comprehensive workflow encompasses transcriptomic dataset processing, age stratification, machine learning, and immune infiltration analysis to identify age-specific hub genes. This computational phase is followed by in vivo experimental validation using an LPS-induced ARDS rat model, employing histological assessment, ELISA, and Western blotting to rigorously evaluate the SEW+FIR intervention and verify hub gene expression. Sixteen pyroptosis-associated genes were identified, among which AXL and GSDME were predominantly associated with ARDS severity in older patients, whereas SPP1 was more relevant in younger individuals. Distinct immune signatures were observed, with M2 macrophage enrichment and immunosuppression in older patients, contrasted by pro-inflammatory activation in younger ones. Functional analyses implicated these genes in metabolic, inflammatory, and immune regulatory pathways. In vivo, SEW+FIR treatment alleviated lung injury, suppressed the production of inflammatory cytokines (IL-1β, IL-18, IL-6, TNF-α), and modulated the expression of AXL, GSDME, and SPP1. Collectively, these findings underscore age-dependent differences in pyroptosis-related mechanisms in ARDS and identify AXL, GSDME, and SPP1 as preliminary biomarker candidates that warrant further clinical validation. In the current murine experimental model, SEW+FIR demonstrated protective effects by alleviating systemic inflammation and modulating pyroptosis-related signaling, providing foundational in vivo support for its potential as a non-pharmacological adjunctive strategy.
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