Jove-Journal of Visualized Experiments最新文献

With increasing societal competition, mental health issues among college students have become increasingly prominent, while traditional mental health education often suffers from limited formats and effectiveness. This study investigates the mechanisms and effectiveness of horticultural therapy (HT), a nature-based intervention, in improving college students' mental health using a sequential mixed-methods explanatory design. In the quantitative phase, 112 college students were randomly assigned to an experimental group receiving HT intervention or a control group, with the experimental group participating in eight weekly HT sessions (90 min each) involving activities such as flower arrangement and herbal tea tasting. Intervention effects were assessed using standardized psychological scales and facial expression video analysis. In the qualitative phase, textual data reflecting participants' authentic experiences were collected and analyzed using grounded theory with NVivo 12. Results showed that, compared with the control group, the experimental group demonstrated significantly higher post-intervention scores in meaning in life (p = 0.017), overall affect index (p = 0.044), and life satisfaction (p = 0.046). Within the experimental group, post-intervention levels of meaning in life (p = 0.02), affect balance (p = 0.007), and positive affect (p = 0.019) were significantly higher than pre-intervention values. Facial expression analysis revealed notable differences in the distribution of seven expressions between groups, with positive expressions increasing over time, particularly during flower arranging and tea tasting activities. Qualitative analysis further identified a four-layer pathway model (Environment → Cognition → Emotion → Behavior), explaining 36%, 32%, 16%, and 15% of the variance, respectively. Overall, horticultural therapy effectively enhances college students' life satisfaction, sense of life meaning, and affect balance, thereby improving mental health through a multi-layered psychological pathway.

This protocol provides a methodological framework for conducting a meta-analysis to evaluate the effects of nutritional therapies on the nutritional status of gastric cancer patients. By following this protocol, researchers will be able to systematically identify, appraise, and synthesize evidence from randomized controlled trials and comparative studies. The steps include: formulating a comprehensive search strategy across multiple databases; dual-independent screening and selection of studies using predefined PICOS criteria (population, interference, comparison, outcome, and study design); standardized data extraction and harmonization of outcome measures; assessment of study quality using the Cochrane Risk of Bias 2.0 tool; and statistical synthesis using appropriate models, with exploration of heterogeneity and sensitivity analyses. Application of this protocol to the available evidence suggests potential benefits of specific nutritional interventions, e.g., oral supplementation for body weight, though high heterogeneity underscores the need for rigorous and standardized methodology in this field. A continuous model with fixed or random effects was used to get the mean difference (MD) with 95% confidence intervals (CIs). A total of 18 studies, involving 3,586 subjects, were selected for the meta-analysis. Oral nutritional supplementation had a significantly increased body weight (MD, 0.74; 95% CI, 0.20-1.27, p = 0.007) compared to the control in patients with gastric cancer. Early enteral nutrition had significantly improved prealbumin levels (MD, 22.53; 95% CI, 13.37-31.69, p < 0.001) compared to parenteral nutrition in patients. However, no significant differences were found between enteral immunonutrition and standard enteral nutrition for albumin (MD, 0.57, 95%CI, -0.31-1.44, p=0.20), prealbumin (MD, 0.23, 95%CI, -0.29-0.76, p=0.38), or transferrin levels (MD, 0.11, 95%CI, -0.09-0.32, p=0.28). The studied data showed that using oral nutritional supplementation had significantly increased body weight compared to control, and early enteral nutrition had significantly improved prealbumin levels compared to parenteral nutrition. However, more studies are required to validate this finding.

Endometrial cancer (EC) ranks among the least prevalent gynecological tumors worldwide, with rising incidence due to aging and obesity. Lymph node metastasis remains common in Uterine Corpus Endometrial Carcinoma (UCEC), necessitating new prognostic biomarkers to guide treatment. In this research, UCEC information from the Cancer Genome Atlas (TCGA) was analyzed, and the results were validated using the Gene Expression Omnibus (GEO). Eighteen differentially expressed genetic factors associated with nicotinamide metabolism (NMRDEGs) were identified. Gene Set Enrichment Analysis (GSEA) indicated their role in oxidative stress, hypoxia, glycolysis, and apoptosis processes. Univariate Cox regression identified six key genes (AURKA, CDKN3, FOXM1, CDKN2A, TK1, and CDK1), utilized to progress a hazard prediction framework. Protein-protein interaction (PPI) analysis uncovered additional hub genes such as CDK2, CCNA2, TP53, and FOXM1. The six key genes showed strong prognostic value, and the study's risk model could guide clinical decisions. Nicotinamide metabolism was found to be significantly linked with EC progression. This study offers new perceptions into the role of nicotinamide metabolism in EC and suggests possible avenues for treatment advancements.

Most neurobiological studies are conducted on laboratory rodents. Despite many similarities across mammalian brains, important differences also exist, which can be misleading in translation. Marked interspecies differences have been found in brain plasticity, particularly neurogenesis. Different neurogenic processes can be prevalent because of evolutionary tradeoffs, displaying variation across brain structures and mammals and shifting the potential for plasticity in divergent species, such as mice and humans. Comparing widely different species raises multiple issues: comparative studies encounter technical difficulties when large-sized brains are involved; the use of heterogeneous experimental approaches by different laboratories can limit the comparison of results; heterogeneity may be related to different time courses of neurodevelopmental processes across mammals, thus adding variables to the comparison. To tackle these limitations, an approach to study the interspecies variation of a population of layer II cortical immature neurons in mammals widely differing for brain size, gyrencephaly, socioecological niche, and age was established. Despite some variables that cannot be fully standardized, a method that combines reduced heterogeneity in collecting brains and the establishment of common anatomical structures as reference points for performing the cell counting on corresponding brain levels is proposed. Data obtained (e.g., cell densities) can be mapped onto phylogenetic trees to reveal evolutionary patterns and analyzed for covariance with neuroanatomical features (e.g., brain size, cortical surface area). This approach has demonstrated remarkable variation in the number of cortical immature neurons between phylogenetic groups and uncovered covariation with brain size. The method can also be used to quantify differences through different developmental stages in the same species and can be extended to other diverse mammals and biological processes to map comparable results that allow for more accurate quantification of different cell populations in adult brains to support plasticity.

Diabetes mellitus (DM) is a widespread chronic metabolic illness around the world, and prolonged hyperglycemia not only causes a variety of problems but also affects metabolic and immunological activities, compromising patients' nutritional status. The Listen-Establish-Adopt-Reinforce-Name-Strengthen (LEARNS) nursing paradigm, based on a patient-centered concept, is a new strategy that aims to foster self-directed learning, improve nurse-patient engagement, increase self-care awareness, and encourage two-way communication. This study looked at the effect of LEARNS nursing on the nutritional status and general health indices of older persons with DM. A total of 168 hospitalized patients aged 60 years or older were involved, with 89 in the LEARNS group and 79 receiving standard treatment. Propensity score matching was used to reduce confounding bias, and the intervention lasted four weeks. After the intervention, patients in the LEARNS group had significantly improved glycemic markers, including reduced HbA1c, FPG, and 2hPG values (P < 0.05). Self-management capacity improved, as evidenced by higher scores on the Chinese Diabetes Management Self-Efficacy Scale. Nutritional indicators such as serum albumin (ALB), hemoglobin (HGB), total serum protein (TSP), and transferrin (TRF) increased, whereas the proportion of high-risk patients identified by the PG-SGA malnutrition score dropped. Quality-of-life assessments also showed improvement, with higher Health-Promoting Lifestyle Profile (HPLP-II) and lower Pittsburgh Sleep Quality Index (PSQI) ratings. The LEARNS group had a considerably higher patient satisfaction rate (92.13%) compared to the control group (P < 0.05). Overall, the data demonstrate that the LEARNS nursing model improves nutritional status, self-management efficacy, and quality of life in senior diabetic patients while also improving glucose control. The study's shortcomings include a single-center design and a short follow-up time. More studies with larger samples and longer periods of observation are needed to confirm long-term effects.

HER2-positive breast cancer is clinically aggressive and frequently disseminates to regional lymph nodes, yet the transcriptomic programs associated with nodal spread remain incompletely defined. Here, we profiled matched tumor-draining lymph nodes (TDLN) and metastatic tumor-involved lymph nodes (TMLN) from six patients with HER2-positive breast cancer using RNA sequencing (RNA-seq). Differential expression analysis was performed with DESeq2, applying Benjamini-Hochberg correction (FDR < 0.05) and an absolute log2 fold-change threshold (|log2FC| ≥ 1). We identified 237 differentially expressed genes (182 upregulated and 55 downregulated in TMLN vs. TDLN). Functional enrichment analyses highlighted extracellular matrix (ECM)-receptor interaction, PI3K-AKT signaling, focal adhesion, and cholesterol metabolism as prominent pathways associated with metastatic lymph nodes, suggesting coordinated ECM remodeling and signaling activation during nodal dissemination. Among the most upregulated genes, POSTN emerged as a key candidate and was further linked to adverse clinicopathologic characteristics. In an independent validation cohort (n = 120), higher POSTN expression was associated with inferior disease-free survival. Collectively, these results nominate POSTN as a potential prognostic biomarker and support ECM-centered and PI3K-AKT-related mechanisms as actionable biological features of lymph-node metastasis in HER2-positive breast cancer.

Surgical intervention has demonstrated established efficacy for benign, borderline, and low-grade malignant tumors of the pancreatic head, as well as for chronic pancreatitis. While conventional surgical approaches are associated with significant trauma and considerable loss of digestive function, the current focus in managing these conditions has shifted toward preserving functional organs through minimally invasive techniques. Through an in-depth understanding of the peripancreatic vascular arches, our team has modified the classic Beger procedure to achieve complete en bloc resection of pancreatic head tissue. By leveraging 3D laparoscopic technology, we have successfully integrated the advantages of minimal invasiveness and precise resection. This refined technique preserves duodenal and biliary integrity while ensuring complete removal of pancreatic head tissue, effectively reducing surgical complications, minimizing operative trauma, and decreasing postoperative gastrointestinal dysfunction. Consequently, patient outcomes are substantially improved. This article aims to elucidate the key technical details of this procedure, enabling more surgeons to master and enhance its success rate, thereby providing better treatment options for eligible patients.

Sepsis is characterized by a dysregulated host immune response and remains a leading cause of mortality worldwide. Long non-coding RNA NEAT1 has been implicated in inflammatory diseases, but its specific role in macrophage polarization during sepsis has not been fully defined. Here, we systematically examine the NEAT1/miR-181a-5p/HMGB1 axis across clinical samples, cultured macrophages, and a CLP mouse model. Quantitative PCR, western blotting, dual-luciferase reporter assays, and RNA pull-down experiments are used to confirm the competitive endogenous RNA (ceRNA) interaction among NEAT1, miR-181a-5p, and HMGB1. Functional assays, including immunofluorescence, transwell migration, and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining, are applied to assess macrophage polarization, migration, and apoptosis. In vivo, the CLP model combined with ELISA and histopathology validates the impact of NEAT1 knockdown on cytokine profiles and organ injury. NEAT1 and HMGB1 are upregulated, whereas miR-181a-5p is downregulated, in patients with sepsis and in lipopolysaccharide-stimulated macrophages. Silencing NEAT1 promotes M2 macrophage polarization, reduces pro-inflammatory cytokines, impairs macrophage migration, and alleviates tissue damage in septic mice via the miR-181a-5p/HMGB1 axis. To our knowledge, this is the first integrated protocol to characterize the lncRNA-microRNA-HMGB1 regulatory circuit in sepsis using harmonized clinical, in vitro, and in vivo approaches. It provides a methodological framework for targeting NEAT1-related ceRNA networks as potential therapeutic strategies.

Stable isotope methods for estimating vitamin A total body stores (TBS) quantitatively are based on the principle of isotope dilution. Briefly, a single oral dose ²H- or ¹³C-labeled vitamin A is administered to an individual, and plasma concentrations of labeled and unlabeled retinol are measured by mass spectrometry at a pre-specified time after dosing, usually 14-21 days. TBS is calculated using the retinol isotope dilution (RID) equation or the mass balance equation. The RID and mass balance equations require information on retinol specific activity in plasma (SAp; i.e., tracer-to-tracee ratio), which is obtained from mass spectrometry measurements. Both equations require values for coefficients to account for absorption and storage of the oral dose of stable isotope-labeled retinol at the time of TBS estimation. Published values for the coefficients can be used in the equations. Alternatively, TBS can be determined by model-based compartmental analysis of plasma retinol kinetic data using WinSAAM Simulation, Analysis, and Modeling software. Briefly, a super-subject study design with model-based compartmental analysis of plasma retinol kinetic data can be used to determine group TBS and population-specific values for the composite coefficient (FaS), which is subsequently used in the RID equation to determine individual TBS. The super-subject design requires an estimate of mean dietary vitamin A intake for the group of participants, and blood sampling at ~11-16 time points over ~28-91 d, with 5-7 participants/time point, but each participant provides only 2-3 blood samples. Liver vitamin A concentration can be estimated from TBS, using an assumption for the proportion of TBS found in liver and an estimate of liver weight. Vitamin A status is assessed by comparing estimated liver vitamin A concentration with proposed cutoff values for categorizing status across the full continuum, from deficient to excess vitamin A stores.

Brain metastases are a common and devastating complication of advanced solid tumors, frequently associated with poor prognosis, neurological decline, and reduced quality of life. The incidence of central nervous system (CNS) failure and neurological death is rapidly increasing, yet the mechanisms driving the final stages of brain metastasis, such as secondary dissemination, re-colonization, and the contribution of the histological growth pattern (HGP) as a potential surrogate parameter, remain poorly understood. The standardized stereotactic intracortical injection model enables precise and reproducible implantation of tumor cells, or mixed populations including stromal or immune components, directly into the mouse cerebral cortex. This protocol also supports the creation of a preclinical tissue archive, offering a robust platform for investigating essential aspects of CNS colonization, such as: metastatic outgrowth, HGP-specific growth dynamics, and pathophysiological mechanisms contributing to neurological failure. Additionally, this model supports pharmacological testing in a reproducible clinically relevant context. Unlike systemic injection methods (e.g., tail vein or intracardiac), which are optimized for studying early metastatic steps but result in variable and often low rates of brain colonization, the stereotactic model ensures consistent, brain-specific metastatic growth and enables the investigation of late stages of CNS metastasis. Compared to ex vivo systems such as organoids or brain slice cultures, the in vivo stereotactic model preserves vascularization, systemic signaling, and the full complexity of the brain's immune landscape, supporting long-term studies of tumor progression and therapeutic response. By providing a reproducible and clinically relevant platform, our model advances the field's ability to identify prognostic markers, explore therapeutic strategies, and understand the mechanisms of late-stage brain metastasis.