Journal Of Visualized Experiments[JOURNAL]
Sun
200 papers
A Single Intravenous Dose of Indole-producing Attenuated Brucella in NOD Mice: Assessment of Type 1 Diabetes Onset and Islet Immune Remodeling.
Delgado K, Adams LG, de Figueiredo P … +1 more , Song J
J Vis Exp · 2026 May · PMID 42296207 · Publisher ↗
Type 1 diabetes (T1D) results from autoimmune destruction of pancreatic β cells, and current therapies fail to restore durable immune tolerance. Metabolically engineered bacteria producing immunomodulatory metabolites re...
Type 1 diabetes (T1D) results from autoimmune destruction of pancreatic β cells, and current therapies fail to restore durable immune tolerance. Metabolically engineered bacteria producing immunomodulatory metabolites represent a promising single-dose therapeutic strategy. Here, this paper presents a comprehensive protocol in which a single intravenous administration of an indole-producing attenuated Brucella melitensis strain BmΔvjbR::tnaA is used to assess effects on the onset of autoimmune diabetes in prediabetic female NOD/ShiLtJ mice. The protocol begins with preparing BmΔvjbR::tnaA from frozen glycerol stocks, growing it in selective media to mid-log phase, and calculating colony-forming units based on optical density. Female NOD mice, <5-week-old are restrained without anesthesia and injected intravenously via lateral tail vein with a defined bacterial dose in phosphate-buffered saline while maintained on heating pads. Blood glucose is monitored 2x weekly after a 2 h fast for 70-105 days, with diabetes defined as glucose ≥250 mg/dL on two consecutive readings. At experimental endpoints, harvested pancreas are fixed in neutral-buffered formalin for 72 h, paraffin-embedded, and sectioned for hematoxylin and eosin staining, insulin immunohistochemistry, and total islet area quantification. Formalin-fixed paraffin-embedded sections are submitted for spatial proteomics with a 25-marker antibody panel to visualize regulatory and effector immune populations. Finally, fresh pancreatic tissue undergoes single-cell isolation, 3' library preparation, sequencing, and Seurat-based bioinformatics analysis to profile transcriptional changes across immune and stromal populations. This integrated protocol enables reproducible single-dose bacterial therapy delivery and multi-modal characterization of islet immune remodeling, providing a framework for testing metabolite-engineered microbes in autoimmune models and dissecting tissue-specific immune mechanisms at single-cell resolution.
Causal Effects of Dietary Macronutrients and Micronutrients on Myopia and Other Major Ocular Disorders: A Mendelian Randomization Study.
Xu Z, Li M
J Vis Exp · 2026 May · PMID 42296206 · Publisher ↗
Myopia and other major ocular disorders, such as cataracts, glaucoma, and age-related macular degeneration (AMD), pose growing public health concerns worldwide. While nutrition is believed to influence ocular development...
Myopia and other major ocular disorders, such as cataracts, glaucoma, and age-related macular degeneration (AMD), pose growing public health concerns worldwide. While nutrition is believed to influence ocular development and degeneration, causal evidence remains limited. A two-sample Mendelian randomization (MR) analysis was performed using large-scale genome-wide association study (GWAS) summary data to evaluate the causal effects of dietary macronutrient and micronutrient intake on the risk of myopia and other ocular diseases. The inverse variance-weighted (IVW) method was used as the primary analysis, supported by weighted median, MR-Egger, and MR-PRESSO for sensitivity testing. To account for multiple comparisons, false discovery rate (FDR) correction was applied. After FDR adjustment, higher genetically predicted vitamin C levels remained significantly associated with an increased risk of AMD (odds ratio [OR] = 1.324, 95% confidence interval [CI]: 1.133-1.546). Several additional associations were observed at nominal significance levels (P < 0.05), including higher relative dietary fat intake with increased myopia risk (OR = 1.033, 95% CI: 1.003-1.063), sugar intake with reduced cataract risk (OR = 0.746, 95% CI: 0.561-0.993), iron with lower myopia risk (OR = 0.995, 95% CI: 0.991-0.999), vitamin D with increased cataract risk (OR = 1.256, 95% CI: 1.062-1.487), folic acid with reduced cataract risk (OR = 0.843, 95% CI: 0.724-0.981), and phosphorus with reduced glaucoma risk (OR = 0.727, 95% CI: 0.554-0.953). No evidence of horizontal pleiotropy or heterogeneity was observed. These findings suggest a potential causal role of certain nutrients in ocular disease risk, although most associations did not remain significant after multiple testing correction and should therefore be interpreted with caution. Further studies are needed to confirm these findings and explore underlying biological mechanisms.
Transcriptomic Analysis Reveals Mitochondrial-Driven Subgroups in Atopic Dermatitis Lesions.
Yu Y, Shang Y, Ge X … +2 more , Dong L, Li K
J Vis Exp · 2026 May · PMID 42296205 · Publisher ↗
Atopic dermatitis (AD) is a common and chronic inflammatory skin disease with global prevalence. Its clinical heterogeneity and complex molecular mechanisms pose significant challenges for the development of effective th...
Atopic dermatitis (AD) is a common and chronic inflammatory skin disease with global prevalence. Its clinical heterogeneity and complex molecular mechanisms pose significant challenges for the development of effective therapies. To explore AD molecular heterogeneity using lesional skin transcriptomic data, characterize their biological and immune profiles, and identify key genes underlying the differentiation. Differentially expressed genes were identified using DESeq2, followed by pathway and co-expression analyses via GSEA and WGCNA, respectively. Mitochondrial-related genes were extracted by intersecting DEGs and WGCNA modules with the MitoCarta3.0 database, and their functional relevance was assessed through GO and KEGG enrichment. Hub genes were identified by protein-protein interaction network analysis, which were subsequently used to construct a classification model. Transcriptional regulators were predicted using hTFtarget, while immune cell infiltration was quantified using CIBERSORT. Two molecular subgroups were identified. Cluster 1 was enriched in cell signaling and adhesion pathways, whereas Cluster 2 exhibited upregulation of oxidative phosphorylation and proteasome-related processes. A total of 85 mitochondrial-associated genes, primarily involved in energy metabolism, were differentially expressed between clusters. PPI network analysis identified four hub genes (BAD, BOLA1, CHCHD5, and ISOC2), which were significantly upregulated in Cluster 1. A hub gene-based classifier demonstrated strong discriminatory power (area under the curve > 0.7). Predicted key transcriptional regulators included ATF3, BRD2, BRD4, and CEBPA. Immune profiling revealed higher regulatory T cell infiltration in Cluster 1 and increased follicular helper T cells in Cluster 2. This study reveals two molecularly and immunologically distinct AD subtypes, characterized by differential mitochondrial function and immune microenvironment signatures.
Prediction of Chronic Low Back Pain Risk Based on Dietary Trace Elements Using Multiple Machine Learning Models and Dual Interpretability Frameworks.
Zhou L, Wang J, Wang C … +1 more , Xu H
J Vis Exp · 2026 May · PMID 42296204 · Publisher ↗
The relationship between dietary trace elements and the risk of chronic low back pain (LBP) remains unclear. Data from the National Health and Nutrition Examination Survey (NHANES) 2001-2004 and 2009-2010 cycles were ana...
The relationship between dietary trace elements and the risk of chronic low back pain (LBP) remains unclear. Data from the National Health and Nutrition Examination Survey (NHANES) 2001-2004 and 2009-2010 cycles were analyzed. Multicollinearity was assessed using Spearman's correlation and variance inflation factors (VIF), and feature selection was performed using the Boruta algorithm. Six machine learning models were subsequently developed, with SHAP and LIME applied to enhance interpretability and identify key associations between dietary trace elements and LBP risk. Among the evaluated models, Random Forest demonstrated the best predictive performance, both when incorporating demographic variables and when using dietary trace elements alone. Interpretability analyses consistently identified several dietary components-including moisture, theobromine, calcium, caffeine, sodium, and vitamin C-as inversely associated with LBP risk. Although model discrimination was moderate (AUC ≈ 0.60-0.72), these findings reveal clinically relevant patterns that may support population-level risk stratification and highlight potentially modifiable dietary factors for future investigation.
Benzo[a]pyrene and Rheumatoid Arthritis: An Integrated Computational Investigation.
Han Y, Chen G, Li W … +7 more , Wu H, Li J, Yang H, Wang M, Liu Y, Wang J, Zhang Y
J Vis Exp · 2026 May · PMID 42296203 · Publisher ↗
Polycyclic aromatic hydrocarbons (PAHs), ubiquitous environmental pollutants, are considered significant environmental factors contributing to the pathogenesis of RA. Benzo[a]pyrene (BaP), a key component of PAHs, may be...
Polycyclic aromatic hydrocarbons (PAHs), ubiquitous environmental pollutants, are considered significant environmental factors contributing to the pathogenesis of RA. Benzo[a]pyrene (BaP), a key component of PAHs, may be associated with RA onset; however, the underlying toxicological mechanism remains to be fully elucidated. In this study, we systematically addressed this knowledge gap using an integrated approach combining network toxicology, machine learning, and molecular docking. Initially, a network toxicology analysis was conducted based on the molecular structure of BaP. By integrating and screening target information from multiple databases, 15 potential RA-related target genes of BaP were ultimately identified, and their interaction network was constructed. GO and KEGG enrichment analyses revealed that these genes were significantly enriched in biological processes such as leukocyte migration and immune cell signal transduction and were associated with the NF-κB and T cell receptor signaling pathways, among others. Subsequent topological analysis using the STRING database and Cytoscape software screened out five core genes (LCK, ZAP70, ITK, GZMA, and ITGAL), whose importance was further validated through machine learning. Molecular docking and molecular dynamics simulation results indicated that BaP exhibits strong binding affinity for the protein products of these target genes, resulting in the formation of conformationally stable complexes. In summary, this study employs an integrated computational approach to elucidate the potential mechanisms by which BaP may contribute to RA development, thereby offering a theoretical foundation for future investigations into the prevention and treatment of RA associated with environmental pollutants.
Acupuncture-related Digital Joint Infection Managed With Staged Surgery And Oral Antibiotic Therapy.
Bai L, Li J, Zhou W … +2 more , Zou X, Lu H
J Vis Exp · 2026 May · PMID 42296200 · Publisher ↗
Acupuncture is generally considered safe; however, inadequate aseptic technique may result in infections involving the skin, soft tissue, bone, or joints. Gram-negative bacilli causing hand infections are uncommon and ma...
Acupuncture is generally considered safe; however, inadequate aseptic technique may result in infections involving the skin, soft tissue, bone, or joints. Gram-negative bacilli causing hand infections are uncommon and may present diagnostic and therapeutic challenges, particularly when multiple digits are involved. A 73-year-old woman developed progressive pain, swelling, and functional limitation of the right third and left fifth fingers approximately 1 month after acupuncture. Imaging suggested osteomyelitis with involvement of the interphalangeal joint. Separate sterile specimens were obtained from each affected digit before antibiotic initiation. Cultures identified Morganella morganii in the right third finger and Proteus mirabilis in the left fifth finger. Both isolates were susceptible to third-generation cephalosporins, beta-lactam/beta-lactamase inhibitor combinations, carbapenems, and fluoroquinolones, with low minimum inhibitory concentrations supporting levofloxacin use. A staged surgical approach was performed, consisting of initial debridement with placement of antibiotic-loaded bone cement, followed by delayed wound repair after infection control. Systemic antimicrobial therapy was guided by susceptibility testing, transitioning from intravenous ceftriaxone to oral therapy. At discharge, oral levofloxacin (750 mg once daily) was prescribed to complete an approximately 6-week treatment course. At 6-week follow-up, both wounds had completely epithelialized, with resolution of swelling and marked pain reduction. Functional recovery was evident, with substantial improvement in interphalangeal joint range of motion. Inflammatory markers normalized, and follow-up radiographs demonstrated bone consolidation without recurrent infection. At 3 months, no relapse or systemic complications were observed. This case illustrates that, in selected patients with acupuncture-related digital osteoarticular infection, a stepwise approach integrating surgical debridement, microbiological confirmation, and susceptibility-guided antimicrobial therapy may achieve satisfactory short-term outcomes. High-bioavailability oral agents such as levofloxacin may be considered after initial infection control in carefully selected cases; however, this observation is based on a single case and requires validation in larger studies.
Standardized Laser Metal Deposition Protocol For Tribological Evaluation Of Ceramic Modified TI6Al4V Matrix Composites.
Onyeka Franklin O, Walker M, Mphahlele M
J Vis Exp · 2026 May · PMID 42296199 · Publisher ↗
This protocol outlines a laser metal deposition method for creating modified titanium-6Aluminum-4Vanadium (TI6Al4V) matrix composites reinforced with boron carbide and boron nitride to enhance hardness and wear resistanc...
This protocol outlines a laser metal deposition method for creating modified titanium-6Aluminum-4Vanadium (TI6Al4V) matrix composites reinforced with boron carbide and boron nitride to enhance hardness and wear resistance. The approach involves promoting in situ phase formation while retaining partially unmelted ceramic particles for effective reinforcement. The process integrates controlled laser-material interaction to enable chemical reactions and preserve ceramic phases within the titanium matrix. Composite fabrication is achieved using a directed energy deposition (DED) laser metal deposition (LMD) system with a coaxial powder feeding mechanism for uniform powder delivery and enhanced melt pool stability. This method allows for tailored microstructural evolution through reaction-driven reinforcement and particulate strengthening. In contrast to traditional surface engineering techniques, this method offers metallurgical bonding, compositional control during deposition, and microstructural manipulation through energy input adjustment. Composite layers were produced at 1400 W and 2000 W with constant scanning speed and powder feed rate. Microhardness (HV0.5) and ASTM G99 dry sliding wear tests were conducted at loads of 15 N and 25 N. Samples produced at 1400 W exhibited higher average hardness and lower wear volume than those produced at 2000 W. The protocol establishes a standard systematic approach for correlating laser energy density with reinforcement retention, microstructure development, and tribological performance.
The Mammalian Ocular Lens in Focus: Development, Anatomy, Physiology, Transparency, Biomechanics, and Age-related Challenges.
Cheng C, Vu MP
J Vis Exp · 2026 May · PMID 42296198 · Publisher ↗
The lens is a transparent, ellipsoid organ in the anterior chamber of the eye that focuses light onto the retina. Composed of epithelial cells and fiber cells, this 3-dimensional tissue is highly organized to ensure a cl...
The lens is a transparent, ellipsoid organ in the anterior chamber of the eye that focuses light onto the retina. Composed of epithelial cells and fiber cells, this 3-dimensional tissue is highly organized to ensure a clear light path and utilizes multiple strategies to establish and maintain a high refractive index and transparency. In humans, the lens undergoes shape changes to focus light from different distances in a process known as accommodation, and these elastic and biomechanical properties of the lens rely on tissue, cellular, and molecular mechanisms. Cataracts, defined as any opacity in the normally transparent lens, remain the leading cause of blindness in the world. Presbyopia, due to age-related changes in lens stiffness, has been linked to accommodative decline and the need for reading glasses. This review will explore mammalian lens development and lifelong growth, crystallins and cytoskeletal proteins, cell-cell communication and microcirculation, lens biomechanics, and the challenges and novel methods for studying this unique tissue.
CRISPR/Cas9-mediated Endogenous Fluorescent Tagging of Germline-specific Genes in Caenorhabditis elegans.
Cheng L, Lu X, Sun X … +4 more , Wen Z, Zhang X, Miao L, Wang P
J Vis Exp · 2026 May · PMID 42296196 · Publisher ↗
Protein localization in the Caenorhabditis elegans (C. elegans) germline is central to interpreting gene function during gametogenesis, yet conventional transgene approaches often yield variable expression and can be sil...
Protein localization in the Caenorhabditis elegans (C. elegans) germline is central to interpreting gene function during gametogenesis, yet conventional transgene approaches often yield variable expression and can be silenced in germ cells. Here, a practical CRISPR/Cas9 workflow inserting a fluorescent tag into an endogenous locus is described, enabling the generation of stable knock-in alleles that report protein distribution under native regulation. The protocol covers key stages of the procedure: selecting a tagging strategy appropriate for the target protein, delivering CRISPR reagents by gonadal microinjection into young adult hermaphrodites, and recovering injected animals for screening. Knock-in candidates are identified through PCR-based genotyping across two generations to isolate homozygous worms and verify the edited allele. Finally, confocal microscopy is used to verify germline fluorescence and assess subcellular localization in vivo. The workflow is designed to be reproducible and broadly applicable to germline-enriched genes, providing a straightforward route to establish homozygous tagged strains for developmental and cell-biological analyses.
Exosomes: Roles and Applications in Chronic Pain Treatment.
Zhu YZ, Huang SJ, Liu YY
J Vis Exp · 2026 May · PMID 42296195 · Publisher ↗
Chronic pain is increasingly recognized as a maladaptive outcome of dysregulated intercellular communication across neural, immune, and stromal systems. Among emerging mediators, exosomes-nanoscale extracellular vesicles...
Chronic pain is increasingly recognized as a maladaptive outcome of dysregulated intercellular communication across neural, immune, and stromal systems. Among emerging mediators, exosomes-nanoscale extracellular vesicles carrying proteins, lipids, and regulatory RNAs-have attracted considerable attention not only as biomarkers and drug carriers but also as active regulators of pain pathophysiology and potential targets for therapeutic intervention. This review aims to synthesize current knowledge on the roles and applications of exosomes in chronic pain treatment and to highlight their potential clinical relevance. Specifically, the review examines how exosomes contribute to chronic pain in representative conditions, including osteoarthritis, ovarian cancer, and hepatocellular carcinoma. Three major functional dimensions are discussed: mechanistic regulation of pain-related processes, therapeutic modulation through endogenous repair and exogenous interventions, and diagnostic utility based on disease- and stage-specific exosomal signatures. In addition, current challenges and future prospects for the clinical application of exosomes are considered. This review provides an overview of the emerging roles of exosomes in chronic pain and offers insights into their potential use in future pain management strategies.
Network Pharmacology and Rat Caudal-disc Puncture Validation of Yi Qi Huo Xue Decoction For Lumbar Disc Degeneration.
Chai Y, Wang Z, Shen X … +9 more , Wang Z, Li Y, Xu Y, Dai Y, Lin S, Lu Y, Sun Y, Zhang X, Liu J
J Vis Exp · 2026 May · PMID 42296193 · Publisher ↗
In this study, network pharmacology, molecular docking, molecular dynamics simulation, and experimental validation were used to investigate the pharmacological basis and potential molecular mechanisms of Yi Qi Huo Xue De...
In this study, network pharmacology, molecular docking, molecular dynamics simulation, and experimental validation were used to investigate the pharmacological basis and potential molecular mechanisms of Yi Qi Huo Xue Decoction (YQHXD) in lumbar disc degeneration (LDD). A total of 678 active compound-related targets of YQHXD and 296 potential LDD-associated therapeutic targets were identified. Protein-protein interaction network analysis and topological analysis identified several candidate targets, including AKT1, IL6, TNF, BCL2, and STAT3. Enrichment analyses using Gene Ontology and Kyoto Encyclopedia of Genes and Genomes indicated that the candidate targets were linked to functions including protein binding and ATP binding, with pathway enrichment that included PI3K-AKT signaling. Molecular docking predicted interactions between selected active compounds, such as quercetin, Tetraneurin A, and isorhamnetin, and the core targets. Molecular dynamics simulations further suggested that the lowest-energy complexes remained stable. Seventy male Sprague-Dawley rats were randomly assigned to the control, sham-operated, model, positive control, and low-, medium-, and high-dose YQHXD groups. After the establishment of a coccygeal intervertebral disc puncture model in the model, positive control, and YQHXD groups, rats were administered YQHXD or celecoxib by oral gavage for 8 weeks, whereas the control and sham-operated groups received normal saline. Histopathological evaluation showed that YQHXD treatment was associated with reduced intervertebral disc degeneration in this model. Enzyme-linked immunosorbent assay, polymerase chain reaction, and western blot analyses showed changes in the mRNA and protein expression levels of selected targets, including AKT1, BCL2, and STAT3. These findings suggest that YQHXD may exert preclinical protective effects in LDD through multi-component and multi-target regulation involving inflammation- and apoptosis-related mechanisms. This study provides preliminary preclinical evidence supporting further investigation of YQHXD for LDD.
Region-Specific Lens Insertion Strategies For In vivo Calcium Imaging In Mice Across Cortical And Hippocampal Circuits.
Shin J, Yi JH, Jung MW
J Vis Exp · 2026 May · PMID 42296186 · Publisher ↗
In vivo calcium imaging of neuronal populations using surgically implanted optical lenses is widely used to monitor neuronal population activity during behavior. However, surgical success and imaging quality depend criti...
In vivo calcium imaging of neuronal populations using surgically implanted optical lenses is widely used to monitor neuronal population activity during behavior. However, surgical success and imaging quality depend critically on anatomical constraints that vary across brain regions. This protocol provides region-specific surgical guidance for the selection and implantation of imaging lenses in commonly targeted cortical and hippocampal areas. Specifically, it describes a surgical workflow encompassing target localization and skull leveling, region-specific craniotomy, and, when required, dura removal or aspiration of overlying tissue, as well as the selection, insertion, and fixation of appropriate lenses and holders. In particular, we describe three implantation strategies: conventional lens implantation for the ventral hippocampus, lens implantation with tissue aspiration for the dorsal hippocampus, and prism lens implantation for cortical imaging. We further provide practical guidance on how differences in tissue depth, vascular architecture, and circuit organization inform key surgical parameters, including lens diameter, craniotomy size, the extent of tissue removal, insertion speed, hemostatic procedures, and fixation methods. By explicitly documenting surgical considerations that are often underreported in written methods, this protocol aims to reduce trial-and-error during lens implantation, and improve experimental reproducibility. Overall, this protocol provides a practical guide for selecting and reliably implementing region-appropriate lens implantation strategies across cortical and hippocampal circuits.
Lentiviral SgRNA Delivery for CRISPR/Cas9 Editing in Cre-Dependent Cas9 Knock-in Primary Mouse Hepatocytes.
Zhang L, Zhang Y, Chi Y … +3 more , Ren Y, Zhao Q, Zhao D
J Vis Exp · 2026 May · PMID 42296184 · Publisher ↗
Over the past few decades, CRISPR/Cas9-mediated genome editing has transformed functional studies in cell line models by making genetic manipulation highly efficient and feasible. However, extending this technology to pr...
Over the past few decades, CRISPR/Cas9-mediated genome editing has transformed functional studies in cell line models by making genetic manipulation highly efficient and feasible. However, extending this technology to primary hepatocytes remains a major challenge. While cultured primary hepatocytes are indispensable for disease modeling and drug development as they retain key metabolic functions absent in cell lines, their limited in vitro lifespan and negligible proliferative capacity pose fundamental barriers to efficient genome editing. Here, we describe an effective two-step perfusion protocol that enables the isolation of primary hepatocytes with high cell viability (>90%) and high yield (approximately 1 × 10 hepatocytes per adult mouse). Following isolation, lentiviral sgRNA transduction is typically performed within 3-4 h, and genome editing outcomes are assessed 5-7 days post-infection. Using hepatocytes from transgenic LSL-Cas9-EGFP mice, in which the Cas9 cassette was activated by a lentiviral vector co-expressing Cre recombinase and sgRNA, achieving up to 80% allele-level gene knockout efficiency in monolayer cultures. In addition, we achieved approximately 12% gene KO efficiency in three-dimensional hepatocyte organoids (HEOs), which more closely recapitulate the architecture and functional characteristics of native liver tissue. In this protocol, a successful experiment is defined by three criteria: sufficient hepatocyte yield (>5 × 10 cells per mouse) with high viability (>85%), efficient single-guide RNA (sgRNA) delivery into hepatocytes, and validated target gene disruption at the genomic level. This protocol demonstrates the feasibility of efficient in vitro genome manipulation in both monolayer-cultured hepatocytes and HEOs, providing a robust platform for genetic modeling and functional studies of liver diseases.
Investigating The Bidirectional Relationship Between Low Back Pain, Hip Pain, Knee Pain, And Gait Abnormalities: A Mendelian Randomization Study.
Zhou J, Xie X, Zhang K … +7 more , Xia Q, Zhang J, Zhang J, Xin Z, Yuan F, Liu L, Li B
J Vis Exp · 2026 May · PMID 42296183 · Publisher ↗
Gait abnormalities are associated with altered lower-limb biomechanics and functional impairment in musculoskeletal disorders. Although low back pain, hip pain, and knee pain have been linked to impaired gait in observat...
Gait abnormalities are associated with altered lower-limb biomechanics and functional impairment in musculoskeletal disorders. Although low back pain, hip pain, and knee pain have been linked to impaired gait in observational studies, their causal relationships remain unclear. We conducted a bidirectional Mendelian randomization (MR) study to assess potential causal associations between site-specific pain and gait abnormalities. Independent single nucleotide polymorphisms from large-scale genome-wide association studies were used as genetic instruments for low back pain, hip pain, and knee pain. Gait abnormalities, defined in the source GWAS as self-reported difficulty walking, were used as the outcome in forward MR analyses and as the exposure in reverse MR analyses. The inverse-variance weighted (IVW) method was the primary analysis, supported by sensitivity analyses for heterogeneity, horizontal pleiotropy, and robustness. In forward analyses, genetically predicted low back pain (OR = 1.53, 95% CI = 1.058-2.219, P = 0.024) and hip pain (OR = 1.55, 95% CI = 1.067-2.252, P = 0.021) were associated with increased risk of gait abnormalities in IVW analyses. However, the four additional MR methods were not statistically significant, although estimates were generally directionally consistent. Genetically predicted knee pain showed no significant association (OR = 2.15, 95% CI = 0.234-19.789, P = 0.498), with substantial uncertainty reflected by the wide confidence interval. Reverse MR analyses found no evidence that genetic liability to gait abnormalities increased the risk of low back, hip, or knee pain, although only four gait-abnormality SNPs were available. No substantial heterogeneity or horizontal pleiotropy was detected. Overall, this study provides limited IVW-based genetic evidence that low back pain and hip pain may be associated with gait abnormalities. Findings should be interpreted cautiously and validated using larger GWAS datasets with refined phenotypes.
Development of A Cell-Free COPII Vesicle Reconstitution Protocol For Investigating STING Sorting In A HEK-293 Cell-Based System.
Nan Y, Liu M, Yang M … +1 more , Ma W
J Vis Exp · 2026 May · PMID 42296182 · Publisher ↗
The cGAS-STING pathway is a central component of innate immunity. Upon activation, the endoplasmic reticulum (ER)-resident protein STING is packaged into COPII vesicles and transported to the Golgi apparatus to initiate...
The cGAS-STING pathway is a central component of innate immunity. Upon activation, the endoplasmic reticulum (ER)-resident protein STING is packaged into COPII vesicles and transported to the Golgi apparatus to initiate downstream signaling. Despite the importance of this process, the molecular mechanisms governing STING sorting into COPII vesicles remain incompletely understood. Here, an optimized workflow is presented for the in vitro reconstitution of COPII vesicles to facilitate controlled analysis of cargo selection. A scalable method is described for preparing high-concentration cytosol (30-40 mg/mL) from HEK-293F suspension cells, combined with a straightforward procedure for generating semi-permeabilized HEK-293T cells as a defined membrane source. Compared to traditional adherent cell-based systems, this approach improves scalability, reduces cost, and enhances reproducibility. Using STING as a model cargo, COPII vesicle budding and cargo incorporation are validated by Western blot analysis, with a basal packaging efficiency of approximately 20% under defined conditions. This system allows controlled manipulation of biochemical parameters, including nucleotide stimulation, to assess their effects on cargo incorporation. This protocol provides a robust and adaptable platform for investigating COPII-mediated cargo sorting and can be extended to study additional transmembrane proteins and regulatory factors involved in ER export.
Research on Pathological Voice Recognition Based on XGBoost.
Wang L, Chen H, Gong X … +7 more , Shi Y, Lu Z, Zhang L, Chen X, Chen D, Zhou H, Cheng L
J Vis Exp · 2026 May · PMID 42296181 · Publisher ↗
With the continuous growth of human social communication, the number of people suffering from voice disorders is also increasing. Due to the objective and non-invasive advantages of acoustic detection methods for patholo...
With the continuous growth of human social communication, the number of people suffering from voice disorders is also increasing. Due to the objective and non-invasive advantages of acoustic detection methods for pathological voice, the use of speech signal analysis for pathological voice recognition has become a research hotspot. This article first selected 101 continuous vowels /a/ from the German SVD database as the research object. Secondly, using wavelet packet technology for time-frequency analysis, four nonlinear dynamic parameters, namely approximate entropy, sample entropy, fuzzy entropy, and permutation entropy, are extracted from the sub signals as the feature parameter set for the pathological voice classifier. Finally, the machine learning algorithm XGBoost is selected as the pattern recognition method to establish a pathological voice classifier, and the classification performance is verified using five fold cross validation and ROC curve. Experimental results have shown that the accuracy of XGBoost's pathological voice classifier is 0.857, the F1 score is 0.875, and the AUC value is 0.944, all of which are higher than the classifier constructed by SVM, indicating that XGBoost has better performance in pathological voice recognition.
Enhancing Patient-Centered Home Palliative Care for Heart Failure: A Concept Analysis to Guide Nursing Practice and Care Coordination.
Zhou L, Song Y, Chen S … +4 more , Li L, Jiang Y, Zeng X, Zhu Y
J Vis Exp · 2026 May · PMID 42296175 · Publisher ↗
To identify and describe components of the home-based palliative care model for patients with heart failure, re-conceptualize the process, and update the existing model to inform nursing research and clinical practice. H...
To identify and describe components of the home-based palliative care model for patients with heart failure, re-conceptualize the process, and update the existing model to inform nursing research and clinical practice. Heart failure, as a chronic progressive disease, has a continuously increasing incidence and medical burden, and requires multidimensional clinical management strategies. Although palliative care is recommended for improving the quality of life of heart failure patients, its utilization rate is still low, and the core components and implementation pathways of home-based palliative care are not yet clear. Using Rodgers' Evolutionary method, a concept analysis was conducted to define the core components for home-based palliative care in heart failure. Data were drawn from professional literature spanning 1990 to 2025, using the terms "home-based palliative care," "home palliative care," "home palliative treatment," "family hospice care," and "heart failure," "cardiac insufficiency," "congestive heart failure". Abstracts from 101 articles were initially reviewed, with 22 articles retained for analysis. Core concepts were identified, defined, and synthesized. The PRISMA 2020 checklist was used. The refined model clarifies nurses' roles in linking home-based palliative care core attributes: self-care, collaborative palliative care, communication of care goals, and support. It presents patients', family caregivers', and healthcare workers' impact on home-based palliative care triggering, implementation, and outcomes, emphasizing patient-centered continuous evaluation/adjustment. This model provides a structured framework to improve heart failure home care delivery and enhance patient/family quality of life, with potential to inform home palliative care approaches for other chronic illnesses. Further refinement/adaptation is needed to ensure relevance across medical contexts.
Impact of Short and Long Daytime Nap Opportunities on Agility, Aerobic, and Anaerobic Performance in Collegiate Badminton Players.
Li Y, Wu J, Li X
J Vis Exp · 2026 May · PMID 42296172 · Publisher ↗
This study investigated the effects of two daytime nap opportunities (40 min and 90 min) on agility, anaerobic performance, and aerobic capacity in collegiate badminton players. Twenty collegiate badminton players partic...
This study investigated the effects of two daytime nap opportunities (40 min and 90 min) on agility, anaerobic performance, and aerobic capacity in collegiate badminton players. Twenty collegiate badminton players participated in a randomized crossover repeated-measures design, including 10 females (age: 20.9 years ± 1.5 years, height: 167.9 cm ± 4.3 cm, weight: 58.3 kg ± 6.1 kg) and 10 males (age: 21.4 years ± 1.4 years, height: 179.7 cm ± 2.8 cm, weight: 73.0 kg ± 3.4 kg). Three experimental conditions were included: no nap (N0), a 40 min nap opportunity (N40), and a 90 min nap opportunity (N90), with a 72 h washout period between sessions. Following each condition, participants completed tests of agility (Y-test), anaerobic capacity (300 yard Shuttle), and aerobic capacity (Yo-Yo Intermittent Recovery Test). Significant main effects of nap condition were observed for all performance outcomes (p < 0.001). Compared with N0, both N40 and N90 improved agility and anaerobic performance. While both nap durations significantly improved physical performance compared with the no-nap condition, the improvements observed in N90 compared to N40 were statistically significant but characterized by small effect sizes (d = 0.12-0.16). This suggests that while a 90 min nap may offer additional benefits, the incremental gain over a 40 min nap is modest for specific performance outcomes.
Chinese Medicine Enemas for Ulcerative Colitis: Mechanisms and Optimal Duration from a Systematic Review and Meta-Analysis.
Chong J, Chen Y, Jiang F
J Vis Exp · 2026 May · PMID 42296171 · Publisher ↗
This systematic review and meta-analysis aimed to elucidate the mechanisms underlying the efficacy of Chinese medicine enema formulations in animal models of ulcerative colitis (UC) and to provide a scientific basis for...
This systematic review and meta-analysis aimed to elucidate the mechanisms underlying the efficacy of Chinese medicine enema formulations in animal models of ulcerative colitis (UC) and to provide a scientific basis for their potential clinical application. Comprehensive searches of PubMed, Web of Science, Cochrane Library, Elton B. Stephens Company (EBSCO), and ScienceDirect identified relevant animal studies published up to December 2024. Methodological quality was assessed using the SYRCLE risk of bias tool, and statistical analyses were conducted with RevMan 5.3 and Stata 15. Twenty studies comprising 980 animals were included, with quality scores ranging from 2 to 6. Chinese medicine enemas significantly reduced histological colitis score, disease activity index, spleen index, colon damage score, and inflammatory mediators including IL-6, TNF-α, IL-1β, IL-8, COX-2 mRNA, and myeloperoxidase. They also improved body weight change, colon length, intestinal tight junction proteins (ZO-1, Occludin), IL-10 expression, and regulatory T-cell levels (CD4⁺CD25⁺FOXP3⁺), while modulating gut microbiota composition, particularly Firmicutes and Bacteroides. These findings suggest that the therapeutic benefits of Chinese medicine enemas may derive from anti-inflammatory activity, enhancement of intestinal barrier integrity, and regulation of gut flora. However, substantial heterogeneity-likely due to differences in animal species, sex, and intervention duration-and generally low methodological quality limit the strength of the evidence. Further well-designed, high-quality preclinical and clinical studies are warranted to confirm efficacy, optimize treatment protocols, and evaluate safety for human application.
An Integrated Method for Photothrombotic Stroke Modeling and In Vivo Optrode Recording of Neuronal and Astrocytic Activity in Behaving Mice.
Chen S, Zhang W, Huang Z … +7 more , Zhang J, Huang W, Zheng Y, Ming K, Yu L, Yi W, Tang X
J Vis Exp · 2026 May · PMID 42296168 · Publisher ↗
Investigating astrocyte-neuron dynamics following ischemic stroke is essential for understanding post-stroke recovery mechanisms. However, current methodologies often fail to capture real-time interactions between neuron...
Investigating astrocyte-neuron dynamics following ischemic stroke is essential for understanding post-stroke recovery mechanisms. However, current methodologies often fail to capture real-time interactions between neurons and astrocytes in animals executing specific behavioral tasks, limiting our ability to investigate the acute phase of stroke pathology. This protocol presents an integrated method that combines photothrombotic stroke modeling with simultaneous multichannel electrophysiology recording and fiber photometry in awake, behaving mice using a custom-fabricated optrode. The protocol includes focal ischemia induction via photothrombosis followed by simultaneous recording of neuronal spikes and astrocytic calcium transients. The optrode enables concurrent delivery of photothrombosis, calcium signal recording, and optogenetic manipulation without requiring separate surgical procedures. Representative results validate the success in simultaneous recording of astrocytic calcium signal and neuronal spiking. Optogenetic manipulation of astrocytes produces measurable changes in neuronal firing patterns (reduction in firing frequency of pyramidal neurons by 1.55 ± 0.45 Hz and interneuron by 3.64 ± 1.37 Hz compared to pre-optogenetic stimulation, n = 2), confirming that the system is capable of investigating astrocyte-neuron interactions. This integrated approach addresses critical gaps in stroke research methodology by providing real-time, multimodal recordings from the acute to chronic stage of stroke in behaving animals.
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