Journal Of Visualized Experiments[JOURNAL]
Sun
200 papers
A Standardized Protocol for Inducing Stress-Induced Premature Senescence in Primary Human Melanocytes Using Tert-butyl Hydroperoxide.
Hassan A, Guerrero-Navarro L, Martic I … +2 more , Declercq L, Cavinato M
J Vis Exp · 2026 May · PMID 42296165 · Publisher ↗
Environmental stressors, including UV radiation, air pollution, or cigarette smoke, drive extrinsic skin aging. Exposure to these extrinsic aging factors promotes the accumulation of senescent cells, which contributes to...
Environmental stressors, including UV radiation, air pollution, or cigarette smoke, drive extrinsic skin aging. Exposure to these extrinsic aging factors promotes the accumulation of senescent cells, which contributes to the formation of wrinkles and pigmentation disorders. Melanocytes, which provide primary UV protection, exhibit reduced activity with age. Senescent melanocytes accumulate in aged skin and contribute to its aging appearance; However, there are few models to study melanocyte senescence, and this work aims to develop a reproducible model for further research. In this study, we present a senescence model of human neonatal melanocytes (HNM) using tert-butyl hydroperoxide (tBHP), a well-characterized oxidative stress inducer. tBHP disrupts redox homeostasis by depleting cellular antioxidant defenses and promoting radical overproduction, resulting in DNA and protein damage. We characterize the senescence phenotype of tBHP-treated melanocytes through the analysis of established senescence markers, including restricted growth potential, morphological changes, and SA-β-Galactosidase activity. This model provides a valuable tool for investigating stress-induced premature senescence in melanocytes, offering insight into their role in extrinsic skin aging and pigmentation disorders.
Integrated Protocol for Assessing Meningeal Lymphatics: Cisterna Magna Injection, Lymph Node Imaging, Meningeal Dissection, and Whole-Mount Staining.
Zhang B, Wang R, Zhao S … +3 more , Qi L, Li X, Wei B
J Vis Exp · 2026 May · PMID 42296163 · Publisher ↗
Meningeal lymphatic vessels (MLVs) have emerged as critical regulators of central nervous system (CNS) homeostasis, and MLVs dysfunction has been implicated in various neurological disorders. A comprehensive assessment o...
Meningeal lymphatic vessels (MLVs) have emerged as critical regulators of central nervous system (CNS) homeostasis, and MLVs dysfunction has been implicated in various neurological disorders. A comprehensive assessment of MLVs' morphology and function is therefore essential for understanding their role in CNS physiology and disease. This protocol describes an integrated method for evaluating MLVs in mice using four key techniques: Intra-cisterna-magna (i.c.m.) injection of fluorescent tracers, live imaging of cervical lymph nodes (CLNs), meningeal dissection, and whole-mount staining. First, the 30 G needle is connected to the syringe and infusion pump via a polyethylene (PE) tube. After the mouse is anesthetized, the dura mater is exposed by blunt dissection of the neck muscles, and the injection needle is inserted into the cisterna magna (CM) to deliver the tracer into the cerebrospinal fluid (CSF) with precise control. Tracer drainage to the CLNs can be visualized in real time, while subsequent meningeal dissection and whole-mount staining enable detailed morphological analysis of the dorsal MLVs. The entire procedure can be completed within 3 h, encompassing i.c.m. injection, tissue perfusion, skull isolation, and placement in fixative solution. This protocol can also be adapted for intracranial drug delivery or combined with molecular analyses, as the meningeal dissection procedure is compatible with fresh, unfixed tissues suitable for RNA and protein extraction.
Evaluating the Effect of Coated Filters and Pre-Filters on the Infectivity and Concentration of Aerosolized Phi6 Virus.
Salokas J, Sofieva-Rios S, Oksanen LM … +6 more , Sanmark E, Malmgren R, Kuula J, Purhonen R, Romantschuk M, Atanasova NS
J Vis Exp · 2026 May · PMID 42296153 · Publisher ↗
Development of efficient indoor air filtration technologies has gained prominence following the coronavirus disease 2019 (COVID-19) pandemic. The goal of this study is to evaluate the anti-viral efficacy of various coppe...
Development of efficient indoor air filtration technologies has gained prominence following the coronavirus disease 2019 (COVID-19) pandemic. The goal of this study is to evaluate the anti-viral efficacy of various copper and silver-based surface coatings for air filters using a standardized bioaerosol chamber and the enveloped bacteriophage Phi6 as a surrogate for respiratory viruses. Surface testing confirmed that all coatings exhibited anti-viral activity, with the specific copper-based coating formulation (B4) achieving complete inactivation of Phi6 virus within 10 min, A6 in 25 min, and VS-B in 60 min; early reductions in infectivity were primarily due to droplet desiccation. Aerosol experiments showed that high-efficiency particulate air (HEPA) filters and pre-filters substantially reduced viral infectivity even without coatings, with mechanical filtration causing a 1.0-2.0 log reduction, and high-efficiency grade GF-2 glass fiber filters eliminating detectable infectious virus. Coatings did not significantly enhance filtration efficiency, likely due to limited virus contact with coated surfaces. Used GF-2 filters demonstrated improved virus removal, attributed to particle accumulation and filter densification, while coatings did not alter the copy number of viruses passing through. VS-B coated filters exhibited lipophilic properties, suggesting additional functional benefits. This methodology highlights that while filter efficiency is primarily driven by mechanical entrapment and particle accumulation, the integrated surface coatings provide a self-sanitizing mechanism by inactivating trapped viral particles; the integration of ultraviolet (UV) light is proposed as a potential future research direction to enhance system efficacy further.
Development of a Severe Blast Lung Injury Model in a Goat for Ultrasonographic Evaluation.
Xue Y, Zhang H, Du J … +3 more , Wen D, Zhang A, Zeng L
J Vis Exp · 2026 May · PMID 42296151 · Publisher ↗
Blast lung injury (BLI) causes severe lung damage and high morbidity, especially in military and industrial settings, leading to hemorrhage and potential respiratory failure, highlighting the need for improved diagnostic...
Blast lung injury (BLI) causes severe lung damage and high morbidity, especially in military and industrial settings, leading to hemorrhage and potential respiratory failure, highlighting the need for improved diagnostics and treatments. This study aimed to develop a standardized severe BLI model in goats to evaluate the accuracy and feasibility of point-of-care ultrasound for dynamic assessment. Goats were subjected to controlled blast overpressure using the biologic shock tube (BST-Ⅰ) at driving pressures of 4.0 MPa (n = 4), 4.5 MPa (n = 12), and 5.0 MPa (n = 4), respectively. Key parameters, such as peak overpressure, were recorded. Vital signs, lung ultrasound scoring (LUS), oxygenation index (PaO₂/FiO₂), and extravascular lung water (EVLW) were monitored at baseline (0 h pre-injury) and 0.5 h, 3 h, 6 h, 9 h, 12 h post-injury. Thoracic computed tomography (CT) at 0 h and 12 h quantified lung injury ratio, and gross examination post-euthanasia assessed pulmonary hemorrhage and injury score. At 4.5 MPa, the peak overpressure was 396.92 kPa, with a 41.67% mortality rate post-injury, whereas at 4.0 MPa, the mortality rate was 0%. LUS increased over time, showing a negative correlation with PaO₂/FiO₂ and a positive correlation with EVLW at 3 h, 6 h, and 9 h, and a positive correlation with lung injury ratio at 12 h. Gross lung injury area ratio was 42.14% in the 4.5 MPa group, indicating severe injury, while 4.0 MPa showed moderate injury. The 4.5 MPa model was suitable for studying severe injury, unlike 4.0 MPa, which caused only moderate injury, and 5.0 MPa resulted in 100% mortality. A reproducible goat model of blast lung injury was established, effectively using LUS to non-invasively assess pulmonary damage over time, providing a basis for monitoring and exploring therapeutic strategies for acute lung injury.
Trabeculotome Tunneling Trabeculoplasty (3T): A Step-by-step Surgical Protocol for Primary Open-angle Glaucoma.
Qiu Q, He CZ, Wang SZ … +6 more , Yu P, He Y, Fan HY, Jin L, Zeng LZ, Wang NL
J Vis Exp · 2026 May · PMID 42296147 · Publisher ↗
Primary open-angle glaucoma (POAG) requires sustained intraocular pressure (IOP) reduction to delay disease progression. Traditional filtering surgeries, while effective, are associated with significant tissue trauma, po...
Primary open-angle glaucoma (POAG) requires sustained intraocular pressure (IOP) reduction to delay disease progression. Traditional filtering surgeries, while effective, are associated with significant tissue trauma, postoperative complications, and long-term scarring. Minimally invasive glaucoma surgery (MIGS) offers a safer alternative but may be limited by incomplete IOP reduction or long-term structural instability of Schlemm's canal (SC). Trabeculotome tunneling trabeculoplasty (3T) is a novel MIGS technique designed to restore physiological aqueous outflow while preserving trabecular meshwork (TM) structure. The procedure consists of three key steps: trabecular fenestration using a trabeculotome, viscoelastic-mediated dilation of SC, and placement of a tension suture to maintain long-term canal patency. This article provides a step-by-step surgical protocol and accompanying training video for the 3T procedure. Representative outcomes from a single-center, non-comparative case series demonstrate that 3T is associated with significant IOP reduction, decreased dependence on antiglaucoma medications, and a low complication rate. This protocol aims to facilitate standardized clinical implementation of 3T and provide a practical educational resource for surgeons performing minimally invasive glaucoma procedures.
Red Blood Cell Distribution Width and Stent Length Predict In-Stent Restenosis After Myocardial Infarction: A Retrospective Cohort Study.
Qiuyu F, Jianing C, Jiamian L … +1 more , Yanli Z
J Vis Exp · 2026 May · PMID 42296144 · Publisher ↗
This study investigated the predictive value of red blood cell distribution width (RDW) stent length for in-stent restenosis (ISR) drug-eluting stent implantation in patients with acute myocardial infarction. Reliable pr...
This study investigated the predictive value of red blood cell distribution width (RDW) stent length for in-stent restenosis (ISR) drug-eluting stent implantation in patients with acute myocardial infarction. Reliable predictors of restenosis remain limited, highlighting the need for accessible clinical markers. A total of 170 patients who underwent stent implantation were included and classified into restenosis and nonrestenosis groups based on follow-up coronary angiography at 12 months. Clinical characteristics and laboratory parameters were compared between groups. Multivariate logistic regression and receiver operating characteristic analyses were performed to evaluate associations and predictive performance. The results showed that RDW and stent length were significantly higher in patients with restenosis and were independent predictors of restenosis risk. The combined use of these variables demonstrated improved predictive performance compared to using the variables individually. In conclusion, RDW and stent length are significant predictors of ISR and may serve as practical indicators for risk stratification in patients undergoing stent implantation.
Isolation, Synthesis, and Characterization of CD47- and Integrin α4/β1-Modified Macrophage Membrane-Coated Poly(lactic-co-glycolic acid) Nanoparticles.
Cai Z, Li Y
J Vis Exp · 2026 May · PMID 42296142 · Publisher ↗
Atherosclerosis is a chronic inflammatory disease and a major contributor to cardiovascular morbidity and mortality. Targeted delivery of anti-inflammatory agents to atherosclerotic sites remains a significant challenge....
Atherosclerosis is a chronic inflammatory disease and a major contributor to cardiovascular morbidity and mortality. Targeted delivery of anti-inflammatory agents to atherosclerotic sites remains a significant challenge. Herein, we present a detailed protocol for the preparation and characterization of CD47- and integrin α4/β1-co-modified macrophage membrane-coated colchicine-loaded poly(lactic-co-glycolic acid) nanoparticles (MMM/COL NPs). The protocol includes macrophage modification via plasmid transfection and endothelin-1 stimulation, membrane isolation, NP fabrication, and membrane coating. The resulting MMM/COL NPs are evaluated using physicochemical characterization, protein expression analysis, and in vitro and in vivo assays. Functional validation includes assessment of cellular association with activated endothelial cells, macrophage uptake, anti-inflammatory effects in foam cells, and biodistribution in a murine atherosclerosis model. Additionally, key experimental parameters and quality control steps are highlighted to ensure reproducibility and consistency across laboratories. This protocol provides a robust and scalable approach for generating biomimetic NPs for targeted drug delivery applications in atherosclerosis research.
Efficacy of Somatostatin Combined with Omeprazole for Non-Variceal Gastrointestinal Bleeding.
Wang Q, Jiang T, Mao Q … +3 more , Jiang S, Li A, Zhu Q
J Vis Exp · 2026 May · PMID 42296138 · Publisher ↗
This study presents a structured, reproducible retrospective workflow for evaluating combination pharmacological therapy in patients with non-variceal upper gastrointestinal bleeding (NVUGIB) using electronic medical rec...
This study presents a structured, reproducible retrospective workflow for evaluating combination pharmacological therapy in patients with non-variceal upper gastrointestinal bleeding (NVUGIB) using electronic medical record (EMR) data. The goal of this protocol is to provide a standardized framework for extracting, processing, and analyzing retrospective clinical data to assess treatment outcomes. The workflow includes database querying, application of inclusion and exclusion criteria, group allocation based on treatment records, and systematic extraction of predefined clinical and laboratory outcomes. Data are analyzed using a stepwise statistical approach to generate representative outcomes. Representative results demonstrate that combination therapy with somatostatin and omeprazole is associated with improved haemostatic outcomes, reduced rebleeding rates, and enhanced clinical recovery compared with omeprazole alone, without an increase in adverse events. These findings are hypothesis-generating and should be interpreted as association-based observations. This protocol emphasizes reproducibility and can be applied across institutions to evaluate pharmacological interventions using retrospective clinical datasets. The method provides a practical framework for clinicians and researchers to assess treatment effectiveness in real-world settings and may be adapted to other therapeutic areas requiring retrospective outcome evaluation.
Transtail Artery Hepatic Artery Catheterization In Rats Under Direct Vision: A Radiation-Free Method For Establishing Animal Models.
Fu Y, Yao L, Lei S … +6 more , Zhang B, Yang H, Zheng Z, Zhang D, Yang L, Ji Z
J Vis Exp · 2026 May · PMID 42296124 · Publisher ↗
Rat liver tumor models are indispensable for studying trans-arterial chemoembolization (TACE), drug delivery, and tumor biology. However, conventional techniques utilizing fluoroscopy-guided carotid or femoral access are...
Rat liver tumor models are indispensable for studying trans-arterial chemoembolization (TACE), drug delivery, and tumor biology. However, conventional techniques utilizing fluoroscopy-guided carotid or femoral access are often constrained by high equipment costs, technical complexity, and cumulative radiation risks to operators. This protocol presents a radiation-free alternative using open-laparotomy-guided transtail artery catheterization. We provide a comprehensive, step-by-step description of the procedure, beginning with standardized anesthesia and exposure of the ventral caudal artery (VCA). The core of the technique involves performing a midline laparotomy to enable direct visualization of the hepatoduodenal anatomy, allowing for the precise advancement of a 1.6 Fr microcatheter into the common hepatic artery (CHA) or proper hepatic artery (PHA) using a 0.014-inch microguidewire. Our validation study in 32 rats demonstrates that this method achieves a high technical success rate of 96.9% (31/32) with a mean procedure time of 22.5 ± 2.3 min. This technique provides an accessible, reproducible, and transparent platform for laboratories focusing on localized therapeutic evaluation and embolic biology. This method effectively lowers the barrier for establishing high-quality interventional models in preclinical research environments where advanced imaging is unavailable.
Quantification of Changes in Bifidobacteria and Lactobacilli Colony-forming Units During Rifaximin Treatment and Synbiotic Gel in Healthy Volunteers.
Cassís-Nosthas ML, Vorácková FV, Dzib FT … +2 more , Delgadillo AT, Castillo-Martínez L
J Vis Exp · 2026 May · PMID 42296114 · Publisher ↗
Although the gut microbiota is generally stable, it can be influenced by antibiotic treatment. Rifaximin, which is used to treat gastrointestinal disorders, is frequently administered with probiotics to reduce the risk o...
Although the gut microbiota is generally stable, it can be influenced by antibiotic treatment. Rifaximin, which is used to treat gastrointestinal disorders, is frequently administered with probiotics to reduce the risk of dysbiosis. Lactobacillus acidophilus interacts with intestinal cells and uses prebiotic compounds. Bifidobacteria, particularly Bifidobacterium infantis ssp. lactis Bi07, is a probiotic that has been associated with beneficial effects in the gut. In vitro studies have demonstrated the superiority of agave fructans over chicory inulin. No studies have documented changes in fecal lactobacilli and bifidobacteria counts following the concomitant administration of rifaximin and synbiotic. Our objective was to evaluate changes in the CFU counts of gut bifidobacteria and lactobacilli in healthy volunteers after consumption of Synbiotic Gel, which contains Lactobacillus acidophilus NCFM, Bifidobacterium lactis Bi07, and agave inulin as a prebiotic, during 1 week of rifaximin treatment and for 4 weeks after. In a randomized, placebo-controlled, double-blind clinical trial, healthy participants without antimicrobial treatment in the previous month were randomized 1:1 in two groups (10 participants per group). The Synbiotic Gel group received 10.32 g of agave inulin as a prebiotic and 4 × 10 CFU/g of Lactobacillus acidophilus NCFM and 4 × 10 CFU/g of Bifidobacterium lactis Bi07 per day, for 5 weeks, and 200 mg rifaximin twice a day for the first week. The Placebo Gel group received placebo gel for 5 weeks and 200 mg rifaximin twice a day for the first week. Bacterial changes were estimated by quantifying CFU by microbiological assessment of feces. Five weeks later, the lactobacilli and bifidobacteria populations did not change. The detection of Lactobacillus acidophilus NCFM and Bifidobacterium lactis Bi07 in some stool samples suggests transient and partial colonization, highlighting the need for dose-response studies with this synbiotic product.
Assessment-Based Pulp And Dentin Removal Protocols For Vital Pulp Therapy.
Yi JK
J Vis Exp · 2026 May · PMID 42296113 · Publisher ↗
Pulpal pathology is a progressive process that may be reversible in early stages if vital tissue remains and pulpal irritants are eliminated. Therefore, vital pulp therapy should aim to preserve pulp vitality by minimizi...
Pulpal pathology is a progressive process that may be reversible in early stages if vital tissue remains and pulpal irritants are eliminated. Therefore, vital pulp therapy should aim to preserve pulp vitality by minimizing unnecessary tissue removal. In conventional pulpotomy, the extent of pulp resection is predetermined (e.g., partial or full), and stepwise pulpal evaluation is not performed, which may lead to removal of viable tissue. Here, we present a protocol for assessment-based removal of dentin and pulp using Minimized Dentin Removal (MDR) and Minimized Pulp Resection (MPR). The MPR technique selectively removes the pathologically altered superficial layer of exposed pulp while preserving deeper viable tissue. MDR removes caries indicator-stained dentin while maintaining dentin adjacent to the pulp to support underlying tissue preservation. The protocol is applied iteratively, with continuous assessment guiding the extent of tissue removal until a clinically acceptable pulpal surface and dentin free of indicator staining are achieved. In contrast to conventional approaches that rely on predetermined resection levels, this method incorporates real-time evaluation to guide intervention. Overall, the MDR-MPR protocol provides a structured, minimally invasive approach to dentin and pulp management and may support preservation of pulp vitality.
A Stage-Phased Psychodynamic Intervention For School Refusal In Adolescent Bipolar Disorder.
Dong B, Xian Z
J Vis Exp · 2026 May · PMID 42296112 · Publisher ↗
Adolescent bipolar disorder is a severe psychiatric condition frequently complicated by school refusal behavior, a debilitating functional impairment that often persists even after the pharmacological stabilization of mo...
Adolescent bipolar disorder is a severe psychiatric condition frequently complicated by school refusal behavior, a debilitating functional impairment that often persists even after the pharmacological stabilization of mood symptoms. While medication effectively targets biological dysregulation, it frequently fails to resolve the deep-seated separation-individuation conflicts and pathological family dynamics, such as triangulation, that actively maintain social withdrawal. Given the scarcity of manualized psychotherapeutic protocols for this specific clinical challenge, this article demonstrates a structured, stage-phased psychodynamic psychotherapy protocol for the treatment of adolescent bipolar disorder comorbid with severe school refusal behavior. We present a comprehensive 60-session intervention applied to a 17-year-old female patient. The protocol integrates standard pharmacotherapy with a three-phase psychodynamic model: (1) Alliance and stabilization; (2) Working through, utilizing dream analysis and transference interpretation; and (3) Termination and separation. Progress was monitored via serial electroencephalogram (EEG) topography and standardized psychological scales. Application of this protocol was associated with the resolution of school refusal and successful university entry in this patient. Longitudinal assessment demonstrated a significant reduction in depression scores to within normal ranges, although anxiety scores remained above the clinical threshold at follow-up. Furthermore, while EEG topography demonstrated overall stability, mild -band dysregulation persisted. This protocol provides a step-by-step roadmap for clinicians, highlighting that resolving underlying family triangulation may be a valuable component in supporting successful school reintegration in pediatric bipolar disorder.
SET Net A Hybrid Deep Learning Framework For EEG Based Attention Relaxation Classification In Brain Computer Interface Applications.
Janapati R, Maram B, Saidhbi S … +2 more , Desai U, Nayak P
J Vis Exp · 2026 May · PMID 42296109 · Publisher ↗
Brain Computer Interface (BCI) technology is used as a tool to provide a direct communication channel between the human brain and the external environment. This has applications in assistive and interactive systems. The...
Brain Computer Interface (BCI) technology is used as a tool to provide a direct communication channel between the human brain and the external environment. This has applications in assistive and interactive systems. The non-invasive acquisition of neural activity using Electroencephalography (EEG) is standard with BCI systems. However, it is difficult to effectively classify cognitive states, including attention and relaxation, because the EEG signal is non-stationary and noisy. To overcome these shortcomings, this study proposes a new hybrid deep learning architecture namely, squeeze-and-excitation (SE) transformer network (SET-Net). In this method, EEG signals are preprocessed and divided into temporal windows to analyze the significant spectrogram representations. The proposed model achieves a classification accuracy of 93.7%, with F1-score of 0.93 and ROC-AUC of 0.98. This demonstrates the effectiveness of the hybrid deep learning model with enhanced discrimination of EEG-based attention-relaxation state. This offers a scalable system for real-time BCI applications as assistive technology in human-computer interaction.
Preparation of Conspecific Alarm Cue From Fathead Minnows.
Minichiello G, Holloway V, Johnson J … +6 more , Rumley E, Sambugaro G, Steinberg K, Turner L, Yake A, Ward J
J Vis Exp · 2026 May · PMID 42296090 · Publisher ↗
Chemical alarm cues are powerful tools in behavioral research, particularly in studies of fish within the superorder Ostariophysi. In nature, injured fish release these cues into the water, where they are innately recogn...
Chemical alarm cues are powerful tools in behavioral research, particularly in studies of fish within the superorder Ostariophysi. In nature, injured fish release these cues into the water, where they are innately recognized by conspecifics and trigger fear responses such as freezing and hiding. Despite their widespread use, methods for the preparation of alarm cues lack consistency. Even within studies on the same species, preparation techniques and concentration often differ, making it difficult to generalize results across experiments. Here, a standardized protocol is described for preparing and administering alarm cues in a widely used model species, the fathead minnow (Pimephales promelas). To acquire the alarm cue, epithelial tissue samples were dissected from euthanized P. promelas females and homogenized, and then the resulting cue was diluted and filtered. Data are presented from a behavioral experiment comparing refugial use by 14 days post hatch (dph) larvae in the presence versus absence of these olfactory cues to confirm that fish detect and recognize the cues. This procedure is intended to promote consistency in chemical ecology research and facilitate more robust comparisons across studies of alarm cue-mediated behavior.
Examination of the Utility of Ultrasound in Prenatal Diagnosis and Perinatal Outcomes for 13 Instances of Mirror Syndrome.
Xu L, Wang J
J Vis Exp · 2026 May · PMID 42296089 · Publisher ↗
This study investigated the diagnostic utility of prenatal ultrasound in identifying mirror syndrome, drawing on the existing literature. A comprehensive series of assessments was undertaken on fetal growth and developme...
This study investigated the diagnostic utility of prenatal ultrasound in identifying mirror syndrome, drawing on the existing literature. A comprehensive series of assessments was undertaken on fetal growth and development, organogenesis, and ancillary structures, including amniotic fluid and placenta, using systematic prenatal ultrasonography in conjunction with hemodynamic evaluations. In cases where fetal edema was identified, additional chromosomal karyotyping and chromosomal microarray analyses were recommended. An analysis of 13 patients diagnosed with mirror syndrome and admitted to the Women's Hospital, Zhejiang University School of Medicine, from January 2020 to July 2025 was conducted. The medical information of these patients was examined, encompassing clinical characteristics, evaluation of fetal and fetal appendage status, laboratory analyses, and perinatal outcomes. The prevalent maternal clinical manifestations included bilateral lower limb edema (100%), hypertension (84.6%), anemia (69.2%), and proteinuria (38.4%). Almost all cases presented with decreased hematocrit and serum albumin levels. In the study, 13 instances exhibited placental thickness > 4 cm, while eleven instances presented placental thickness > 6 cm. Pathological examination of the placenta revealed villous edema in eight cases. Mirror syndrome is rare; however, the fetal prognosis is poor. Early and precise diagnosis of mirror syndrome is essential for improving outcomes for both the mother and the fetus.
Green Synthesis of Magnesium Nanoparticles From Native Neotropical Savannah Plants For Scalable Nanophytocosmetic Formulations.
Gasparetto LS, Novaes MA, Campos CMV … +5 more , Torres JAS, Salentim VHB, Toledo LAS, Cândido ES, Franco OL
J Vis Exp · 2026 May · PMID 42296082 · Publisher ↗
This study describes a protocol for the green synthesis of magnesium-based nanoparticles using ethanol extracts of Stryphnodendron adstringens, a plant species native to the Neotropical savanna, and their incorporation i...
This study describes a protocol for the green synthesis of magnesium-based nanoparticles using ethanol extracts of Stryphnodendron adstringens, a plant species native to the Neotropical savanna, and their incorporation into nanophytocosmetic formulations. The procedure includes collection and preparation of plant material, followed by ethanol extraction, in which phytochemicals act as reducing and stabilizing agents for magnesium ions. Nanoparticles are synthesized under controlled conditions using magnesium chloride and plant extracts, enabling an environmentally friendly process without the use of toxic reducing agents. Nanoparticle properties are characterized using dynamic light scattering and atomic force microscopy to assess size distribution, morphology, and colloidal behavior. The protocol further describes the incorporation of the synthesized nanoparticles into a cosmetic serum formulation. Stability is evaluated under controlled storage conditions by monitoring organoleptic properties, pH, and viscosity. This method provides a reproducible and sustainable approach for producing plant-mediated magnesium nanoparticles and assessing their application in cosmetic formulations.
Establishment of an Acid-Induced Het-1A Cell Model and Functional Analysis of the miR-107/FGFRL1 Axis.
Lin C, Li J, Zheng X … +4 more , Wang X, Wu F, Li C, Hu C
J Vis Exp · 2026 May · PMID 42296079 · Publisher ↗
Extensive evidence suggests that microRNAs (miRNAs) play a key role in gastrointestinal pathophysiological processes. However, their specific mechanisms in reflux esophagitis (RE) remain poorly understood. Here, we prese...
Extensive evidence suggests that microRNAs (miRNAs) play a key role in gastrointestinal pathophysiological processes. However, their specific mechanisms in reflux esophagitis (RE) remain poorly understood. Here, we present a protocol to establish an acid-induced RE model and to investigate the miR-107/FGFRL1 axis using molecular and cellular assays. We conducted clinical sample analyses, including serum collection from patients with R E (n=94) and healthy controls (n=94), followed by quantitative detection via RT-qPCR. In vitro, an acid-induced injury model was established using Het-1A cells exposed to hydrochloric acid. Additionally, functional validation of the cells was performed using MTT cell viability assays and dual-luciferase reporter assays. Representative results indicate that miR-107 was downregulated in the serum of RE patients and in acid-treated Het-1A cells. miR-107 mediated hydrochloric acid-induced changes in Het-1A cell viability and inflammation levels by directly targeting FGFRL1 and is associated with changes in cell viability and inflammation through FGFRL1 targeting. This protocol enables investigation of miR-107/FGFRL1-mediated mechanisms in RE. This approach provides a reproducible platform for studying molecular mechanisms in RE.
The Mechanism of Ermiao Powder In Treating Prostatitis: A Study Based on Network Pharmacology and Experimental Verification.
Li QR, Wang P
J Vis Exp · 2026 May · PMID 42296078 · Publisher ↗
This protocol presents a combined network pharmacology and experimental validation approach to investigate the mechanism of action of Ermiao Powder (EMP) in the treatment of prostatitis. The workflow follows a stepwise d...
This protocol presents a combined network pharmacology and experimental validation approach to investigate the mechanism of action of Ermiao Powder (EMP) in the treatment of prostatitis. The workflow follows a stepwise design: (i) database-driven screening of active compounds and targets, (ii) construction and analysis of interaction networks, and (iii) in vitro experimental validation. First, active ingredients of EMP and their corresponding targets were retrieved from the TCMSP database, while prostatitis-related targets were obtained from the GeneCards database. The intersection of EMP targets and disease targets was used to identify key therapeutic targets. Second, a "Drug-Active Ingredient-Target-Disease" network was constructed using Cytoscape. Protein-protein interaction (PPI) analysis and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment were performed on the key targets, revealing that inflammatory factors such as IL6 and TNF served as core targets, and pathways including PI3K/AKT and MAPK were significantly enriched. Finally, in vitro validation was conducted using an LPS-induced inflammatory model of prostate epithelial cells. Treatment with EMP significantly reduced the release of pro-inflammatory factors IL-6 and TNF-α compared to the LPS-treated group (p < 0.05), confirming the anti-inflammatory efficacy of EMP. This integrative approach provides a reproducible framework that can be adapted to investigate other multi-component therapeutic systems. In conclusion, this study preliminarily elucidates that EMP exerts its anti-prostatitis effects through a multi-component, multi-target, and multi-pathway mechanism, closely associated with the suppression of inflammatory responses, thereby offering a scientific basis for its clinical application.
A Study on An Intelligent Diagnosis and Treatment Assistant System For Acupuncture in Diminished Ovarian Reserve Based on A Knowledge Graph.
Zhu Z, Lu J, Li K … +6 more , Wen J, Huang Y, Lv Z, Li C, Xia Y, Xu T
J Vis Exp · 2026 May · PMID 42296077 · Publisher ↗
Diminished ovarian reserve (DOR) is increasing in incidence and affecting younger women. Acupuncture has been used clinically for DOR; however, practice remains subjective, experience-dependent, and lacks standardized gu...
Diminished ovarian reserve (DOR) is increasing in incidence and affecting younger women. Acupuncture has been used clinically for DOR; however, practice remains subjective, experience-dependent, and lacks standardized guideline implementation. Building on our previous general intelligent acupuncture system, this study targets gynecological diseases and uses DOR as the clinical entry point to develop a specialized knowledge graph and decision-support system. This study included 82 classical acupuncture texts, Chinese and English literature, and clinical data. Knowledge was extracted using a hybrid approach combining large language models and manual annotation to construct a full acupuncture knowledge graph and a DOR-specific gynecology subgraph. The system employs BiLSTM-CRF-based entity recognition and dual-mode retrieval to automate syndrome differentiation and acupuncture prescription generation. The full graph contains 16,558 entities and 80,084 relations, while the gynecology subgraph contains 8,677 entities and 27,092 relations. In a clinical evaluation of 90 patients with DOR, the system achieved a diagnostic agreement rate of 86.7% compared with expert assessments. The mean prescription appropriateness score was 4.65. Inter-rater agreement among experts, measured using Kendall's coefficient of concordance, was W ≈ 0.13 (P < 0.05), indicating statistically significant but limited agreement. These findings suggest that the system can provide structured support for acupuncture-based syndrome differentiation and prescription generation in DOR, with potential as an auxiliary clinical decision-support tool.
Arrhythmias and Coronary Slow Flow: A Potential Pathogenic Link.
Jinhong C, Zongyin W
J Vis Exp · 2026 May · PMID 42296075 · Publisher ↗
Coronary slow flow (CSF) is characterized by recurrent resting angina and delayed distal vessel filling, despite the absence of obstructive coronary artery disease on angiography. With the continuous advancement of coron...
Coronary slow flow (CSF) is characterized by recurrent resting angina and delayed distal vessel filling, despite the absence of obstructive coronary artery disease on angiography. With the continuous advancement of coronary interventional techniques, cardiologists have increasingly recognized CSF as a distinct phenomenon, and its pathogenesis has become a major research focus. Arrhythmias, common cardiovascular conditions, often present with clinical manifestations similar to those of CSF. Moreover, both conditions share certain pathogenic factors and pathophysiological processes, including endothelial dysfunction, inflammation, autonomic imbalance, and microvascular abnormalities. A clearer understanding of these overlapping mechanisms may help clinicians manage patients with CSF more scientifically and comprehensively. The present review summarizes the proposed pathogenic mechanisms of CSF, including microvascular disorders, endothelial dysfunction, inflammation, arrhythmias, anatomical factors, early atherosclerosis, and genetic polymorphisms. It also discusses the possible association between CSF and arrhythmias, highlights current uncertainties regarding causality, and considers how these mechanisms may broaden future therapeutic perspectives for this disease.
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