The role of tertiary lymphoid structure and B cells in nasopharyngeal carcinoma: Based on bioinformatics and experimental verification.

OBJECTIVE: Transcriptomic characteristics and prognosis of tertiary lymphoid structures (TLS) and infiltrating B cells in nasopharyngeal carcinoma (NPC) remain unclear. Here, NPC transcriptomic data and clinical samples were used to investigate the role of infiltrating B cells and TLS in NPC. METHODS: We investigated the gene expression and infiltrating immune cells of NPC patients and further investigated the clinical relevance of B cell and TLS signatures. Transcriptional features of infiltrating B cell subsets were revealed by single-cell RNA sequencing (scRNA-seq) analysis. Immunohistochemical (IHC) and HE staining were performed to validate the clinical relevance of infiltrating B cells and TLS in NPC samples. RESULTS: 27 differentially expressed immune-related genes (IRGs) associated with prognosis were identified, including B cell marker genes CD19 and CD79B. The higher B cells and TLS signature scores were associated with better outcomes and early pathological staging in 88 NPC patients. ScRNA-seq identified five distinct B cell subsets in NPC, including the BC-4 cluster associated with poor outcomes and the BC-0 cluster associated with better outcomes. EBV infection was positively associated with the formation of TLS. Furthermore, experimental results showed that the infiltration of B cells in NPC tissues was higher than that of normal tissues, and the density of TLS in an early stage of NPC was higher than that in advanced-stage TLS. CONCLUSION: Our findings demonstrate the functional importance of distinct B cell subsets in the prognosis of NPC. Additionally, we confirmed that B cells and TLS may serve as prognostic biomarkers of survival for NPC patients.

Bioinformatics Prediction and Experimental Validation of the Role of Macrophage Polarization and Ferroptosis in Gestational Diabetes Mellitus.

Purpose: Gestational diabetes mellitus (GDM) is a common metabolic disorder during pregnancy that is associated with placental inflammation and adverse pregnancy outcomes. However, the mechanisms of inflammation in GDM are still unclear. Methods: Bulk transcriptome, single-cell transcriptome, clinical information, and samples were collected from GSE154414, GSE70493, GSE173193 and a retrospective cohort. Bioinformatics prediction was used to explore the mechanisms of placental inflammation, and multiplex immunofluorescence was used to validate the results. Results: First, we found that GDM is characterized by low-grade inflammation and is linked to several adverse pregnancy outcomes, as supported by our collected clinical data. Additionally, we identified ten hub genes (FCGR3B, CXCR1, MMP9, ITGAX, CCL5, GZMB, S100A8, LCN2, TGFB1, and LTF) as potential therapy targets and confirmed the binding of corresponding predictive therapeutic agents by molecular docking. Transcriptome sequencing analysis has shown that macrophages are primarily responsible for the emergence of placental inflammation, and that M1 macrophage polarization increased while M2 macrophage polarization decreased in GDM when compared to the control sample. Multiplex immunofluorescence staining of CD68, CD80, and ACSL4 was performed and suggested that ferroptosis of macrophages may contribute to placental inflammation in GDM. Conclusion: In conclusion, our findings provide a better understanding of the mechanisms of inflammation in GDM and suggest potential therapeutic targets for this condition.

Predictive Model of Functional Exercise Compliance of Patients with Breast Cancer Based on Decision Tree.

Objective: Regular functional exercise can help recover the functions of upper limb for patients with breast cancer. By finding the influencing factors of functional exercise compliance and constructing a predictive model, patients with a poor functional exercise compliance can be better identified. This study aims to find out the factors influencing the functional exercise compliance of patients with breast cancer and build a predictive model based on decision tree. Methods: Convenience sampling was used at two tertiary hospitals in Shantou from August 2020 to March 2021. Data of patients with breast cancer patient was obtained from questionnaires and based on demographics, Constant-Murley Score, Functional Exercise Compliance Scale for Postoperative Breast Cancer Patients, Champion Health Belief Model Scale, Social Support Rating Scale, Disease Perception Questionnaire and Family Care Index Questionnaire. Possible influencing factors of functional exercise compliance were analyzed using correlation analysis as well as univariate and binary logistic regression analysis through SPSS v25, and a CHAID decision tree was used to construct a predictive model on training, validation and test sets via SPSS Modeler v18 at a ratio of 6:2:2. Prediction accuracy, sensitivity, specificity and AUC were used to analyze the efficacy of the predictive model. Results: A total of 227 valid samples were collected, of which 145 were assessed with a poor compliance (63.9%). According to a logistic regression analysis, perceived benefits, time after surgery and self-efficacy were influencing factors. The prediction accuracy, sensitivity, specificity and AUC of the predictive model, based on a CHAID decision tree analysis, were 70.73%, 57.1%, 77.8% and 0.81 respectively. Conclusion: The predictive model, based on a CHAID decision tree analysis, had a moderate predictive efficacy, which could be used as a clinical auxiliary tool for clinical nurses to predict patients' functional exercise compliance.

The Antimicrobial Peptide Esculentin-1a(1-21)NH2 Stimulates Wound Healing by Promoting Angiogenesis through the PI3K/AKT Pathway.

Delayed wound healing is a persistent medical problem mainly caused by decreased angiogenesis. Esculentin-1a(1-21)NH2 [Esc-1a(1-21)NH2], has broad-spectrum antibacterial properties which comes from frog skins. It has shown promise as a treatment for wound healing. However, its effects on angiogenesis as well as the mechanism by which esc-1a(1-21)NH2 enhanced wound healing remained unclear. In this study, we analyzed the structural properties and biocompatibility of esc-1a(1-21)NH2 and evaluated its effect on wound closure using a full-thickness excision model in mice. Our results showed that esc-1a(1-21)NH2 significantly accelerated wound healing by increasing collagen deposition and angiogenesis, characterized by elevated expression levels of platelet, endothelial cell adhesion molecule-1 (CD31) and proliferating cell nuclear antigen (PCNA). Furthermore, the angiogenic activity of esc-1a(1-21)NH2 was confirmed in vitro by various assays. Esc-1a(1-21)NH2 significantly promoted cell migration and cell proliferation in human umbilical vein vascular endothelial cells (HUVECs) via activation of the phosphatidylinositol 3'-kinase (PI3K)/protein kinase B (AKT) pathway, and upregulated the expression of CD31 at both mRNA and protein levels. The effect of esc-1a(1-21)NH2 on angiogenesis was diminished by LY294002, a PI3K pathway inhibitor. Taken together, this study demonstrates that esc-1a(1-21)NH2 accelerates wound closure in mice by promoting angiogenesis via the PI3K/AKT signaling pathway, suggesting its effective application in the treatment of wound healing.

Emodin accelerates diabetic wound healing by promoting anti-inflammatory macrophage polarization.

Diabetic wound healing, characterized by chronic inflammation, remains a medical challenge. The failure of prompt conversion from pro-inflammatory M1-like macrophage to pro-healing M2-like macrophage is the main obstacle to diabetic wounds. Emodin, an anthraquinone derivative, has multiple bioactivities, including antibacterial, anticancer, and anti-inflammatory. Recently, emodin has shown potential in promoting wound healing. However, the underlying molecular mechanism remains unclear. In this study, we examined the effects of emodin on wound healing in db/db diabetic mice using a full-thickness excision model. Our results showed that emodin can remarkably accelerate healing by enhancing extracellular matrix (ECM) synthesis and granulation tissue formation. We identified 32 potential targets of emodin by network pharmacology analysis, and our transcriptome analysis highlighted the down-regulation of the NF-κB signaling pathway mediated by emodin. Mechanistically, emodin was shown to inhibit the p65-NF-κB complex and promote the proportion of M2 (anti-inflammatory)-like phenotype macrophages both in vitro and vivo. Then, bone-marrow-derived macrophages were co-cultured with fibroblasts (mouse dermal fibroblasts cells). Treatment of emodin significantly increased the proportion of M2-polarized macrophages and the expression level of TGF-ß, a typical ECM formation-related cytokine secreted by the M2 macrophages in the co-cultured supernatant. We further revealed that emodin improved the proliferation of mouse dermal fibroblasts (MDFs) cells and upregulated the expression levels of collagen III, fibronectin and α-SMA in MDFs cells in emodin-treated co-culture systems. 1D11, a neutralizing antibody for all three major TGF-ß isoforms, diminished the biological effects of emodin on proliferation and ECM formation in MDFs cells. Taken together, our study suggests emodin may serve as an effective therapeutic agent for diabetic wounds.

Multimetric structural covariance in first-episode major depressive disorder: a graph theoretical analysis.

BACKGROUND: Abnormalities of cortical morphology have been consistently reported in major depressive disorder (MDD), with widespread focal alterations in cortical thickness, surface area and gyrification. However, it is unclear whether these distributed focal changes disrupt the system-level architecture (topology) of brain morphology in MDD. If present, such a topological disruption might explain the mechanisms that underlie altered cortical morphology in MDD. METHODS: Seventy-six patients with first-episode MDD (33 male, 43 female) and 66 healthy controls (32 male, 34 female) underwent structural MRI scans. We calculated cortical indices, including cortical thickness, surface area and local gyrification index, using FreeSurfer. We constructed morphological covariance networks using the 3 cortical indices separately, and we analyzed the topological properties of these group-level morphological covariance networks using graph theoretical approaches. RESULTS: Topological differences between patients with first-episode MDD and healthy controls were restricted to the thickness-based network. We found a significant decrease in global efficiency but an increase in local efficiency of the left superior frontal gyrus and the right paracentral lobule in patients with first-episode MDD. When we simulated targeted lesions affecting the most highly connected nodes, the thickness-based networks in patients with first-episode MDD disintegrated more rapidly than those in healthy controls. LIMITATIONS: Our sample of patients with first-episode MDD has limited generalizability to patients with chronic and recurrent MDD. CONCLUSION: A systems-level disruption in cortical thickness (but not surface area or gyrification) occurs in patients with first-episode MDD.

Aberrant integrity of the cortico-limbic-striatal circuit in major depressive disorder with suicidal ideation.

BACKGROUND: Suicidal ideation is a common symptom of major depressive disorder (MDD) that reflects a cognitive alteration in the background of intense emotional dysregulation. Amygdala is a critical emotion processing center that facilitates moving from emotional appraisal to action. However, whether MDD patients with suicidal ideation show dysconnectivity of the amygdala within a large-scale neurocognitive circuitry remains unknown. METHODS: Participants were 22 MDD patients without suicidal ideation (MDD-NSI), 59 MDD patients with suicidal ideation (MDD-SI), and 60 healthy controls (HCs). We compared the amygdala-based resting-state functional connectivity of four amygdala subregions across the three groups. We selected brain regions with significant between-group differences in amygdalar conectivity as the regions of interest (ROI) and performed ROI-to-ROI and graph-theoretical analyses to explore dysconnectivity patterns at various granularity levels. RESULTS: Brain regions showing omnibus differences across the three groups were distributed across a cortico-limbic-striatal circuitry. MDD-SI had unique dysconnectivity of the lateral amygdala with caudate, middle temporal gyrus, and postcentral gyrus compared with the other two groups. MDD-SI and MDD-NSI had shared dysconnectivity of the medial amygdala with medial superior frontal gyrus and middle temporal gyrus. Within the derived cortico-limbic-striatal circuitry, MDD-SI exhibited lower global connectivity, reduced sigma (small-worldness), but increased lambda (path-length) than HCs. Reduced sigma correlated with increased severity of suicidal ideation. We achieved high classification accuracy (84.09%, with AUC 0.82) in distinguishing MDD-SI from MDD-NSI. CONCLUSIONS: Aberrant integrity of the cortico-limbic-striatal circuit centered on the amygdala provides a promising neural substrate for suicidal ideation that requires further investigation in MDD.

Decreased Cortical Folding of the Fusiform Gyrus and Its Hypoconnectivity with Sensorimotor Areas in Major Depressive Disorder.

BACKGROUND: Neuroimaging studies have revealed abnormal cortical folding pattern and disruptive functional connectivity in major depressive disorder (MDD). Combining structure and function in the same population may further our understanding of the neuropathological mechanisms of MDD. METHOD: Sixty-two patients with MDD and 61 healthy controls (HCs) underwent structural and resting-state functional magnetic resonance imaging (MRI). Group differences in the cortical folding (measured by local gyrification index (LGI)) were analyzed in FreeSurfer. Taking the brain regions with significant group differences in LGI as seed regions, the resting-state functional connectivity analysis was further conducted to explore the corresponding functional connectivity alterations. RESULTS: Comparing with HCs, patients with MDD showed significantly decreased LGI in the right fusiform gyrus (cohen's d = 0.70). In the seed-based functional connectivity analysis, we found that compared with HCs, patients with MDD showed decreased functional connections between the right fusiform gyrus with sensorimotor areas (precentral and postcentral gyrus) (cohen's d = 1.32) and right superior temporal gyrus (cohen's d = 0.94). LIMITATIONS: Main limitations are the relatively small sample size and the cross-sectional study design. CONCLUSION: Decreased LGI in the right fusiform gyrus, as well as decreased functional connectivity between the right fusiform gyrus and the sensorimotor area and right superior temporal gyrus, appears to play a role in the pathophysiology of MDD.

A qualitative study on the experience of empowerment from the perspectives of breast cancer survivors.

AIM: To describe the experience of empowerment for breast cancer survivors in order to increase quality of life after treatment. DESIGN: We adopted a qualitative design in this study. METHODS: A qualitative descriptive approach guided by interpretive description methodology as the theoretical framework was used in this study. Semi-structured group and individual interviews were conducted with eleven female Chinese breast cancer survivors (mean age 54.18 years, 100% female). RESULTS: Many survivors reported that they continued to experience disease-related discomfort from the physiological aspect, but they use multiple ways to improve quality of life. Breast cancer survivors experienced empowerment after treatment. Empowerment mainly required three components: a belief in good health, capability of self-management and acquisition of a good social support system.

Application of in situ simulation teaching in the training of trainee nurses to respond to emergencies.

BACKGROUND: To explore the effect of the in situ simulation teaching method in the emergency training of trainee nurses. METHODS: A total of 108 trainee nurses from the First Affiliated Hospital of Hainan Medical College were selected, and in situ simulation teaching was employed in emergency training. Following the in situ simulation teaching training, a questionnaire was issued to evaluate CIPP (context evaluation, input evaluation, process evaluation, product evaluation) simulation teaching, clinical thinking ability, nursing team cooperation, and student satisfaction. These data were then collected and statistical analysis was conducted. RESULTS: The response rate of this teaching questionnaire was 100%. After using in situ simulation teaching to instruct trainee nurses in emergencies, the satisfaction rate of the trainee nurses was 94.9%, and the satisfaction rate of the instructor with the trainee nurses was 92.2%. After in-situ simulation teaching, the clinical thinking ability (critical thinking ability, systematic thinking ability, evidence-based thinking ability), teamwork ability (trust, team orientation, support, shared mental model and team leadership), the theoretical and clinical practice ability had been improved. CONCLUSIONS: Most nurses agreed that the in situ simulation teaching method can cultivate clinical thinking and teamwork ability for common emergencies, thereby improving their comprehensive quality and job competence, which is invaluable when responding to emergencies.

Serum exosomes accelerate diabetic wound healing by promoting angiogenesis and ECM formation.

Nonhealing wounds in diabetes remain a global clinical and research challenge. Exosomes are primary mediators of cell paracrine action, which are shown to promote tissue repair and regeneration. In this study, we investigated the effects of serum derived exosomes (Serum-Exos) on diabetic wound healing and its possible mechanisms. Serum-Exos were isolated from blood serum of normal healthy mice and identified by transmission electron microscopy and western blot. The effects of Serum-Exos on diabetic wound healing, fibroblast growth and migration, angiogenesis and extracellular matrix (ECM) formation were investigated. Our results showed that the isolated Serum-Exos exhibited a sphere-shaped morphology with a mean diameter at 150 nm, and expressed classical markers of exosomes including HSP70, TSG101, and CD63. Treatment with Serum-Exos elevated the percentage of wound closure and shortened the time of healing in diabetic mice. Mechanistically, Serum-Exos promoted granulation tissue formation and increased the expression of CD31, fibronectin and collagen-ɑ in diabetic mice. Serum-Exos also promoted the migration of NIH/3T3 cells, which was associated with increased expression levels of PCNA, Ki67, collagen-α and fibronectin. In addition, Serum-Exos enhanced tube formation in human umbilical vein endothelial cells and induced the expression of CD31 at both protein and messenger RNA levels. Collectively, our results suggest that Serum-Exos may facilitate the wound healing in diabetic mice by promoting angiogenesis and ECM formation, and show the potential application in treating diabetic wounds.

Psychological resilience negatively correlates with resting-state brain network flexibility in young healthy adults: a dynamic functional magnetic resonance imaging study.

BACKGROUND: Psychological resilience is an important personality trait whose decrease is associated with many common psychiatric disorders, but the neural mechanisms underlying it remain largely unclear. In this study, we aimed to explore the neural correlates of psychological resilience in healthy adults by investigating its relationship with functional brain network flexibility, a fundamental dynamic feature of brain network defined by switching frequency of its modular community structures. METHODS: Resting-state functional magnetic resonance imaging (fMRI) scans were acquired from 41 healthy adults, whose psychological resilience was quantified by the Connor-Davidson Resilience Scale (CD-RISC). Dynamic functional brain network was constructed for each subject, whose flexibility was calculated at all the global, subnetwork and region-of-interest (ROI) levels. After that, the associations between CD-RISC score and brain network flexibility were assessed at all levels by partial correlations controlling for age, sex, education and head motion. Correlation was also tested between the CD-RISC score and modularity of conventional static brain network for comparative purposes. RESULTS: The CD-RISC score was significant negatively correlated with the brain network flexibility at global level (r=-0.533, P=0.001), and with flexibility of the visual subnetwork at subnetwork level (r=-0.576, corrected P=0.002). Moreover, significant (corrected P<0.05) or trends for (corrected P<0.10) negative correlations were found between the CD-RISC score and flexibilities of a number of visual and default-mode areas at ROI level. Meanwhile, the modularity of static brain network did not reveal significant correlation with CD-RISC score (P>0.05). CONCLUSIONS: Our results suggest that excessive fluctuations of the functional brain community structures during rest may be indicative of a lower psychological resilience, and the visual and default-mode systems may play crucial roles in such relationship. These findings may provide important implications for improving our understanding of the psychological resilience.

Effectiveness evaluation of digital virtual simulation application in teaching of gross anatomy.

INTRODUCTION: Digital virtual simulation (DVS) is increasingly used to supplement the teaching of anatomy. DVS can provide the students with three-dimensional (3D) stereoscopic images and precise structures in anatomy teaching. We investigated the effects of digital virtual simulation (DVS) application in gross anatomy teaching. MATERIALS AND METHODS: Fourth-year medical students (n=120), majoring clinical medicine in Class 2013 from Xiangya School of Medicine, Central South University, were assigned into four classes. Two classes received a traditional teaching method and were the control classes, the others received a new teaching reform method-gross anatomy teaching by DVS, and were the experimental classes. After the reform was carried out, the students in teaching reform classes were proceeded to do a subjective evaluation of the teaching method in order to collect feedback information. RESULTS: Students in the experimental group achieved a mean score of 84.97 (±7.86) compared to 78.96 (±5.78) in the control group, with statistical significance between two the groups (p<0.01). The result of the questionnaire survey showed that the students highly approved this reform in many domains. CONCLUSION: The application of digital virtual simulation could be used to supplement the teaching of gross anatomy, including theoretical and experimental teaching such as specimen dissection.

Early-life lipopolysaccharide exposure potentiates forebrain expression of NLRP3 inflammasome proteins and anxiety-like behavior in adolescent rats.

BACKGROUND: Neonatal inflammation may affect brain development and lead to cognitive and emotional deficits at adolescence and adulthood. The nucleotide binding and oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3) is the core component of NLRP3 inflammasome, which may involve in neuroinflammation. We explored if early-life exposure to the bacterial endotoxin lipopolysaccharide (LPS) could promote the expression of proteins related to NLRP3 inflammasome, including NLRP3, the apoptosis-associated speck-like protein (ASC) and cysteiny aspartate-specific protease (Caspase-1) in the forebrain, and behavioral alteration in adolescent rats. METHODS: Two-week old Sprague Dawley rats were divided into naïve control, vehicle (phosphate buffered saline, PBS) control and LPS (100µg/kg, i.p.) treatment groups. Anxiety and depression-like behaviors were examined around 1month age, with the expression of NLRP3, ASC and Caspase-1 in the prefrontal cortex (PFC) and hippocampus analyzed by means of immunohistochemistry and western blot. RESULTS: LPS-treated rats exhibited anxiety but not depressive-like behavior as indicated by results of open field, elevated plus maze, dark-light box, sucrose preference and forced swimming tests. Increased immunolabeling of NLRP3, ASC and Caspase-1 in neurons and/or microglia occurred in the PFC and hippocampus in LPS-treated adolescents relative to controls, with immunoblot shown elevated levels of these proteins. CONCLUSION: Early-life inflammatory stress promotes the expression of NLRP3 inflammasome proteins in the brain and the occurrence of anxiety-like behavior in adolescent rats.

Cross-cultural verification of the EORTC QLQ-C15-PAL questionnaire in mainland China.

BACKGROUND: Simplified by European Organization for Research and Treatment of Cancer (EORTC) Quality-of-Life Questionnaire Core 30 (QLQ-C30), EORTC Quality-of-Life Questionnaire Core 15 Palliative Care (QLQ-C15-PAL) is specifically applied to evaluating palliative care patients' quality of life. AIM: This study examined cross-cultural adaptability and validity of QLQ-C15-PAL for evaluating quality of life of palliative care patients with advanced cancer in mainland China. PARTICIPANTS AND DESIGN: From May to October 2013, 243 palliative care patients in Tianjin Cancer Hospital completed the EORTC QLQ-C30. We extracted QLQ-C15-PAL data for analysis. Physicians completed the Eastern Cooperative Oncology Group Performance Status score and mental state assessment for each patient. RESULTS: A total of 243 patients completed the study. The compliance rate was high, with missing rate for each item ranging from 0% to 2.1%. In addition to emotional function, the remaining dimensions demonstrated a high reliability (Cronbach's alpha > 0.7). Whether we divided patients into two groups according to their Eastern Cooperative Oncology Group Performance Status or divided patients into three groups according to mental status, both sets of results showed significant differences in QLQ-C15-PAL subscale scores (p < 0.05), indicating that the QLQ-C15-PAL scale could be used to distinguish between the aforementioned subgroups. Overall quality of life was moderately correlated with fatigue (r = -0.406) but weakly correlated with other subscales. The proportion of variance (R(2)) ranged from 0.848 to 0.903, which showed that QLQ-C15-PAL subscale scores explained between 84.8% and 90.3% of the original QLQ-C30 score distribution. CONCLUSION: The Chinese version of the EORTC QLQ-C15-PAL questionnaire has high reliability and validity and is therefore suitable for clinical use in China to determine health-related quality of life in Chinese patients with advanced cancer.