Protocol for the Chinese Real-World Evidence for Acute Spinal Cord Injury (ChiRES) study: a prospective, observational, multicentre cohort study of acute spinal cord injury.

INTRODUCTION: Spinal cord injury (SCI) is a catastrophic event with devastating physical, social and occupational consequences for patients and their families. The number of patients with acute SCI in China continues to grow rapidly, but there have been no large prospective cohort studies of patients with acute SCI. This proposed study aims to establish a multicentre, extensive sample cohort of clinical data and biological samples of patients in China, which would aid the systematisation and standardisation of clinical research and treatment of acute SCI, thus reducing the heavy burden of acute SCI on patients and society. METHODS AND ANALYSIS: The Chinese Real-World Evidence for Acute Spinal Cord Injury (ChiRES) study is an observational, multicentre cohort study of patients with acute SCI admitted to the Qilu Hospital of Shandong University and other participating centres with prospective collection of their clinical data and biological samples. We aim to recruit 2097 patients in this study. Demographics, disease history, emergency intervention information, motor and sensory examinations, surgical information, medication information and rehabilitation evaluation will be recorded. This will facilitate the development of a prediction model for complications and prognosis of patients with acute SCI and an evaluation of the current management of acute SCI. Among these variables, detailed information on surgical treatment will also be used to assess procedures for acute SCI treatment. Outcome measurements, including the International Standard for Neurological Classification of Spinal Cord Injury examinations, the occurrence of complications and death, will be performed repeatedly during follow-up. We will analyse imaging data and blood samples to develop SCI imaging markers and biomarkers. ETHICS AND DISSEMINATION: This study protocol has been approved by the Medical Ethics Committee of the Qilu Hospital of Shandong University and all other participating centres. The findings will be disseminated in peer-reviewed journals and academic conferences.

Itaconate protects ferroptotic neurons by alkylating GPx4 post stroke.

Neuronal ferroptosis plays a key role in neurologic deficits post intracerebral hemorrhage (ICH). However, the endogenous regulation of rescuing ferroptotic neurons is largely unexplored. Here, we analyzed the integrated alteration of metabolomic landscape after ICH using LC-MS and MALDI-TOF/TOF MS, and demonstrated that aconitate decarboxylase 1 (Irg1) and its product itaconate, a derivative of the tricarboxylic acid cycle, were protectively upregulated. Deficiency of Irg1 or depletion of neuronal Irg1 in striatal neurons was shown to exaggerate neuronal loss and behavioral dysfunction in an ICH mouse model using transgenic mice. Administration of 4-Octyl itaconate (4-OI), a cell-permeable itaconate derivative, and neuronal Irg1 overexpression protected neurons in vivo. In addition, itaconate inhibited ferroptosis in cortical neurons derived from mouse and human induced pluripotent stem cells in vitro. Mechanistically, we demonstrated that itaconate alkylated glutathione peroxidase 4 (GPx4) on its cysteine 66 and the modification allosterically enhanced GPx4's enzymatic activity by using a bioorthogonal probe, itaconate-alkyne (ITalk), and a GPx4 activity assay using phosphatidylcholine hydroperoxide. Altogether, our research suggested that Irg1/itaconate-GPx4 axis may be a future therapeutic strategy for protecting neurons from ferroptosis post ICH.

Identification of immune-associated biomarker for predicting lung adenocarcinoma: bioinformatics analysis and experiment verification of PTK6.

BACKGROUND: Abnormal expression of protein tyrosine kinase 6 (PTK6) has been proven to be involved in the development of gynecological tumors. However, its immune-related carcinogenic mechanism in other tumors remains unclear. OBJECTIVE: The aim of this study was to identify PTK6 as a novel prognostic biomarker in pan-cancer, especially in lung adenocarcinoma (LUAD), which is correlated with immune infiltration, and to clarify its clinicopathological and prognostic significance. METHODS: The prognostic value and immune relevance of PTK6 were investigated by using bio-informatics in this study. PTK6 expression was validated in vitro experiments (lung cancer cell lines PC9, NCI-H1975, and HCC827; human normal lung epithelial cells BEAS-2B). Western blot (WB) revealed the PTK6 protein expression in lung cancer cell lines. PTK6 expression was inhibited by Tilfrinib. Colony formation and the Cell Counting Kit-8 (CCK-8) assay were used to detect cell proliferation. The wound healing and trans-well were performed to analyze the cell migration capacity. Then flow cytometry was conducted to evaluate the cell apoptosis. Eventually, the relationship between PTK6 and immune checkpoints was examined. WB was used to estimate the PD-L1 expression at different Tilfrinib doses. RESULTS: PTK6 was an independent predictive factor for LUAD and was substantially expressed in LUAD. Pathological stage was significantly correlated with increased PTK6 expression. In accordance with survival analysis, poor survival rate in LUAD was associated with a high expression level of PTK6. Functional enrichment of the cell cycle and TGF-ß signaling pathway was demonstrated by KEGG and GSEA analysis. Moreover, PTK6 expression considerably associated with immune infiltration in LUAD, as determined by immune analysis. Thus, the result of vitro experiments indicated that cell proliferation and migration were inhibited by the elimination of PTK6. Additionally, PTK6 suppression induced cell apoptosis. Obviously, PD-L1 protein expression level up-regulated while PTK6 was suppressed. CONCLUSION: PTK6 has predictive value for LUAD prognosis, and could up regulated PD-L1.

Surface shape alters perceived material softness.

When a human strokes the surface of an object with his/her finger, the surface shape influences the perceived softness of the object. This study introduced a curved surface softness illusion, which alters the perception of material softness. When a surface with curvature is felt by sliding a finger over it, it feels softer than a flat surface made of the same material. In contrast, a rugged surface is perceived as harder. This illusion indicates that, in addition to mechanical hardness, humans judge an object's softness based on its surface shape.

CD4+CD25+ regulatory T cells ex vivo generated from autologous naïve CD4+ T cells suppress EAE progression.

CD4+CD25+ regulatory T cells (Tregs) play an important role in maintaining immune homeostasis in multiple sclerosis (MS). Hence, we aimed to explore the therapeutic efficacy and safety of adoptive cell therapy (ACT) utilizing induced antigen-specific Tregs in an animal model of MS, that is, in an experimental autoimmune encephalomyelitis (EAE) model. B cells from EAE model that were activated with soluble CD40L were used as antigen-presenting cells (APCs) to induce the differentiation of antigen-specific Tregs from naïve CD4 precursors, and then, a stepwise isolation of CD4+CD25highCD127low Tregs was performed using a flow sorter. All EAE mice were divided into Treg-treated group (2 × 104 cells in 0.2 mL per mouse, n = 14) and sham-treated group (0.2 mL normal saline (NS), n = 20), which were observed daily for clinical assessment, and for abnormal appearance for 6 weeks. Afterward, histological analysis, immunofluorescence and real-time PCR were performed. Compared to sham-treated mice, Treg-treated mice exhibited a significant decrease in disease severity scores and reduced inflammatory infiltration and demyelination in the spinal cord. Additionally, Tregs-treated mice demonstrated higher CCN3 protein and mRNA levels than sham-treated mice. The results of this preclinical study further support the therapeutic potential of this ACT approach in the treatment of MS.

Low back pain-driven inpatient stays in the United States: a nationwide repeated cross-sectional analysis.

BACKGROUND: Low back pain (LBP)-driven inpatient stays are resource-intensive and costly, yet data on contemporary national trends are limited. MATERIALS AND METHODS: This study used repeated cross-sectional analyses through a nationally representative sample (US National Inpatient Sample, 2016-2019). Outcomes included the rate of LBP-driven inpatient stays; the resource utilization (the proportion of receiving surgical treatments and hospital costs) and prognosis (hospital length of stay and the proportion of nonroutine discharge) among LBP-driven inpatient stays. LBP was classified as overall, nonspecific, and specific (i.e. cancer, cauda equina syndrome, vertebral infection, vertebral compression fracture, axial spondyloarthritis, radicular pain, and spinal canal stenosis). Analyses were further stratified by age, sex, and race/ethnicity. RESULTS: 292 987 LBP-driven inpatient stays (weighted number: 1 464 690) were included, with 269 080 (91.8%) of these for specific LBP and 23 907 (8.2%) for nonspecific LBP. The rate of LBP-driven inpatient stays varied a lot across demographic groups and LBP subtypes (e.g. for overall LBP, highest for non-Hispanic White 180.4 vs. lowest for non-Hispanic Asian/Pacific Islander 42.0 per 100 000 population). Between 2016 and 2019, the rate of nonspecific LBP-driven inpatient stays significantly decreased (relative change: 46.9%); however, substantial variations were found within subcategories of specific LBP-significant increases were found for vertebral infection (relative change: 17.2%), vertebral compression fracture (relative change: 13.4%), and spinal canal stenosis (relative change: 19.9%), while a significant decrease was found for radicular pain (relative change: 12.6%). The proportion of receiving surgical treatments also varied a lot (e.g. for overall LBP, highest for non-Hispanic White 74.4% vs. lowest for non-Hispanic Asian/Pacific Islander 62.8%), and significantly decreased between 2016 and 2019 (e.g. for nonspecific LBP, relative change: 28.6%). Variations were also observed for other outcomes. CONCLUSIONS: In the US, the burden of LBP-driven inpatient stays (i.e. rates of LBP-driven inpatient stays, resource utilization, and prognosis among LBP-driven inpatient stays) is enormous. More research is needed to understand why the burden varies considerably according to the LBP subtype (i.e. nonspecific and specific LBP as well as subcategories of specific LBP) and the subpopulation concerned (i.e. stratified by age, sex, and race/ethnicity).

Meta-structure-based graph attention networks.

Due to the ubiquity of graph-structured data, Graph Neural Network (GNN) have been widely used in different tasks and domains and good results have been achieved in tasks such as node classification and link prediction. However, there are still many challenges in representation learning of heterogeneous networks. Existing graph neural network models are partly based on homogeneous graphs, which do not take into account the rich semantic information of nodes and edges due to their different types; And partly based on heterogeneous graphs, which require predefined meta-structures (include meta-paths and meta-graphs) and do not take into account the different effects of different meta-structures on node representation. In this paper, we propose the MS-GAN model, which consists of four parts: graph structure learner, graph structure expander, graph structure filter and graph structure parser. The graph structure learner automatically generates a graph structure consisting of useful meta-paths by selecting and combining the sub-adjacent matrices in the original graph using a 1 × 1 convolution. The graph structure expander further generates a graph structure containing meta-graphs by Hadamard product based on the previous step. The graph structure filterer filters out graph structures that are more effective for downstream classification tasks based on diversity. The graph structure parser assigns different weights to graph structures consisting of different meta-structures by a semantic hierarchical attention. Finally, through experiments on four datasets and meta-structure visualization analysis, it is shown that MS-GAN can automatically generate useful meta-structures and assign different weights to different meta-structures.

Implications of different cell death patterns for prognosis and immunity in lung adenocarcinoma.

In recent years, lung adenocarcinoma (LUAD) has become a focus of attention due to its low response to treatment, poor prognosis, and lack of reliable indicators to predict the progression or therapeutic effect of LUAD. Different cell death patterns play a crucial role in tumor development and are promising for predicting LUAD prognosis. From the TCGA and GEO databases, we obtained bulk transcriptomes, single-cell transcriptomes, and clinical information. Genes in 15 types of cell death were analyzed for cell death index (CDI) signature establishment. The CDI signature using necroptosis + immunologic cell death-related genes was established in the TCGA cohort with the 1-, 2-, 3-, 4- and 5-year AUC values were 0.772, 0.736, 0.723, 0.795, and 0.743, respectively. The prognosis was significantly better in the low CDI group than in the high CDI group. We also investigated the relationship between the CDI signature and clinical variables, published prognosis biomarkers, immune cell infiltration, functional enrichment pathways, and immunity biomarkers. In vitro assay showed that HNRNPF and FGF2 promoted lung cancer cell proliferation and migration and were also involved in cell death. Therefore, as a robust prognosis biomarker, CDI signatures can screen for patients who might benefit from immunotherapy and improve diagnostic accuracy and LUAD patient outcomes.

Traumatic spinal injury-related hospitalizations in the United States, 2016-2019: a retrospective study.

BACKGROUND: Traumatic spinal injury (TSI) is associated with significant fatality and social burden; however, the epidemiology and treatment of patients with TSI in the US remain unclear. MATERIALS AND METHODS: An adult population was selected from the National Inpatient Sample database from 2016 to 2019. TSI incidence was calculated and TSI-related hospitalizations were divided into operative and nonoperative groups according to the treatments received. TSIs were classified as fracture, dislocation, internal organ injury, nerve root injury, or sprain injuries based on their nature. The annual percentage change (APC) was calculated to identify trends. In-hospital deaths were utilized to evaluate the prognosis of different TSIs. RESULTS: Overall, 95 047 adult patients were hospitalized with TSI in the US from 2016 to 2019, with an incidence rate of 48.4 per 100 000 persons in 2019 (95% CI: 46.2-50.6). The total incidence increased with an APC of 1.5% (95% CI: 0.1-3%) from 2016 to 2019. Operative TSI treatment was more common than nonoperative (32.8 vs. 3.8; 95% CI: 32.3-33.2 vs. 3.6-4%). The number of operations increased from 37 555 (95% CI: 34 674-40 436) to 40 460 (95% CI: 37 372-43 548); however, the operative rate only increased for internal organ injury (i.e. spinal cord injury [SCI])-related hospitalizations (APC, 3.6%; 95% CI: 2.8-4.4%). In-hospital mortality was highest among SCI-related hospitalizations, recorded at 3.9% (95% CI: 2.9-5%) and 28% (95% CI: 17.9-38.2%) in the operative and nonoperative groups, respectively. CONCLUSIONS: The estimated incidence of TSI in US adults increased from 2016 to 2019. The number of operations increased; however, the proportion of operations performed on TSI-related hospitalizations did not significantly change. In 2019, SCI was the highest associated mortality TSI, regardless of operative or nonoperative treatment.

Analysis between macrophage-related genes with prognosis and tumor microenvironment in non-small cell lung cancer.

Non-small cell lung cancer (NSCLC) is a highly morbid and fatal disease that exhibits individualized differences in prognosis and drug efficacy. Therefore, understanding the molecular mechanism of the occurrence and progression of lung cancer can improve early diagnosis, treatment and prognosis. Macrophages are a crucial component of the tumor microenvironment (TME) due to their high plasticity and heterogeneity. They play a multifaceted role in tumor initiation and progression. In order to elucidate the pathogenesis of tumor-associated macrophages (TAMs) related genes in NSCLC, transcriptomic sequencing, univariate COX regression, LASSO regression and multivariate COX regression analyses were conducted to identify the 11 genes that have the most significant association with prognosis. These genes include FCRLA, LDHA, LMOD3, MAP3K8, NT5E, PDGFB, S100P, SFXN1, TDRD1, TFAP2A and TUBB6. The risk score (RS) was computed, and all samples were split into high- and low-risk groups based on the median RS. The correlation of RS and 11 genes with macrophages was verified by the CIBERSORT deconvolution algorithm. These above results suggest that the risk score developed in this study can be utilized for predicting patients' prognosis and evaluating their immune infiltration status. This study can serve as a guide for subsequent tumor immunotherapy and gene targeting therapy.

How does the target of green innovation for cleaner production change in management process? Quality targeting and link targeting.

The global economy has accelerated the transition to a green, low-carbon economy. An enterprise's green innovation (GI) is directly related to its capacity for sustainable production as a micro-subject of economic development. This study examined the impact of managerial capacity on enterprise green innovation and changes of green innovation targeting. We used data collected manually from 423 Chinese A-share companies from 2010 to 2017. The effect of various external impact signals was then investigated. This study's findings are as follows: (1) Managerial ability stimulated green enterprise innovation. The marginal effect was 0.0696. While quality targeting has focused more on green invention innovation, managerial capacity significantly improved the marginal impact of green substantial innovation by 0.0375; (2) The clean production link targeting analysis confirmed that enterprises focused on end-of-pipe governance innovation (0.0466), along with new energy innovation (0.0495) rather than energy-saving innovation (-0.0227); (3) The multi-period DDD (Difference in Difference in Difference) model revealed that low-carbon city policy promoted green innovation with a diminishing trend; (4) The voluntary environmental regulation signals, ISO14001 certification, displayed a substitute effect for managerial capacity on enterprise green innovation. This paper provides recommendations, including that enterprises should improve the utilization of new and renewable energy while improving and optimizing production processes. The government should also improve innovation incentive policies and strengthen environmental information disclosure.

Can dexamethasone improve postoperative sleep and postoperative delirium in elderly patients undergoing robot-assisted laparoscopic radical prostatectomy? Protocol for a prospective, randomized, double-blind, controlled study.

BACKGROUND: Perioperative sleep disorders (PSD) are an independent risk factor for postoperative delirium (POD), which is a common complication after surgery. Elderly patients who undergo robot-assisted radical prostatectomy (RARP) often experience perioperative sleep disorders (PSD). Dexamethasone, a medication that works by inhibiting the hypothalamic-pituitary-suprarenal cortical axis, can reduce the negative effects of surgical stress. The objective of this study was to determine whether intravenous administration of dexamethasone at the time of anesthesia induction could improve postoperative sleep quality in elderly patients, thereby indirectly reducing the risk of postoperative cognitive impairment and accelerating postoperative rehabilitation. METHODS/DESIGN: This study is a randomized, double-blind, placebo-controlled trial that was conducted at a single center. A sample size of 116 patients was determined through calculation, and these patients were randomly assigned to either the dexamethasone group (group D, n = 58) or the blank control group (group C, n = 58). On the day of surgery, the anesthesia nurse prepared either diluted dexamethasone or saline in advance, according to the patient's assigned group. The blinded anesthesiologist administered the medication during induction, and a dedicated person followed up with the patient for three consecutive postoperative days. All other aspects of care were managed equally between the two groups. The primary outcome measure was sleep quality, while secondary outcome measures included postoperative sleep time, postoperative delirium (POD), pain scores, and other complications. Relevant test measures were recorded for analysis. DISCUSSION: This study aims to investigate the impact of intravenous dexamethasone on sleep quality and duration of patients undergoing robot-assisted radical prostatectomy (RARP). If the findings of this study protocol are affirmative, it could enhance the sleep quality of elderly patients after surgery, thereby minimizing the risk of postoperative delirium (POD), and providing substantial evidence for the perioperative enhanced recovery management of elderly patients. TRIAL REGISTRATION: Chinese clinical trial registry: ChiCTR2200063488, Registered on 5 October 2022.

TRCA-Net: using TRCA filters to boost the SSVEP classification with convolutional neural network.

Objective.The steady-state visual evoked potential (SSVEP)-based brain-computer interface has received extensive attention in research due to its simple system, less training data, and high information transfer rate. There are currently two prominent methods dominating the classification of SSVEP signals. One is the knowledge-based task-related component analysis (TRCA) method, whose core idea is to find the spatial filters by maximizing the inter-trial covariance. The other is the deep learning-based approach, which directly learns a classification model from data. However, how to integrate the two methods to achieve better performance has not been studied before.Approach.In this study, we develop a novel algorithm named TRCA-Net (TRCA-Net) to enhance SSVEP signal classification, which enjoys the advantages of both the knowledge-based method and the deep model. Specifically, the proposed TRCA-Net first performs TRCA to obtain spatial filters, which extract task-related components of data. Then the TRCA-filtered features from different filters are rearranged as new multi-channel signals for a deep convolutional neural network (CNN) for classification. Introducing the TRCA filters to a deep learning-based approach improves the signal-to-noise ratio of input data, hence benefiting the deep learning model.Main results.We evaluate the performance of TRCA-Net using two publicly available large-scale benchmark datasets, and the results demonstrate the effectiveness of TRCA-Net. Additionally, offline and online experiments separately testing ten and five subjects further validate the robustness of TRCA-Net. Further, we conduct ablation studies on different CNN backbones and demonstrate that our approach can be transplanted into other CNN models to boost their performance.Significance.The proposed approach is believed to have a promising potential for SSVEP classification and promote its practical applications in communication and control. The code is available athttps://github.com/Sungden/TRCA-Net.

How Sagittal Spinal Parameters Predict Anterolateral Spinal Ossification Severity: A Study Based on CT Classification.

STUDY DESIGN: Retrospective observational study. OBJECTIVE: The purpose of this study was to establish a CT classification system of anterolateral spinal ossification and analyze the effects of sagittal spinal parameters on ossification. SUMMARY OF BACKGROUND DATA: Patients with diffuse idiopathic skeletal hyperostosis often present with anterolateral ossification of the spine. Few reports consider anterolateral spinal ossification at each spinal level or explore the influence of sagittal alignment. MATERIALS AND METHOD: One hundred and twenty patients (79 males and 41 females) over 60 years old who underwent whole spinal CT scans from October 2018 to November 2021 were analyzed. Volume rendering technique images were used to assess the degree of anterolateral spinal ossification in each intervertebral space, and a classification system was established. Sagittal parameters, such as thoracic kyphosis, lumbar lordosis (LL), cervical lordosis, sacral slope, and thoracolumbar junction angle of the patients were measured. Multifactor stepwise linear regression analysis and ordered logistic regression analysis were used to study the effects of the sagittal parameters on ossification grades. RESULTS: The median age of the study population was 67 years (63-72 IQR). The new classification system classifies the severity of anterolateral spinal ossification in each intervertebral space into grades 0 to 3 with an intra-observer intraclass correlation coefficient value of 0.909 and inter-observer intraclass correlation coefficient value of 0.900. Multivariate stepwise linear regression analysis showed that age (OR=1.30, P <0.001), weight (OR=1.23, P =0.013), and cervical lordosis (OR=1.19, P <0.001) were significantly correlated with total ossification grade. Multifactor ordered logistic regression analysis showed that there was a statistically significant correlation between cervical lordosis and the ossification grades of each intervertebral space of T1~T4 ( P ≤0.005), between thoracic kyphosis and each intervertebral space in T4~T12 ( P ≤0.019), and between thoracolumbar junction angle and each intervertebral space in T10~L3 ( P ≤0.025). LL was significantly and positively correlated with the ossification grades of each intervertebral space in T7~T11 ( P ≤0.041). LL and sacral slope were significantly negatively correlated with the ossification grades of each intervertebral space in L3~S1 ( P ≤0.047). CONCLUSION: Our new classification scheme demonstrated good accuracy and reliability for the evaluation of the severity of anterolateral spinal ossification. The effect of sagittal parameters on anterolateral spinal ossification was position-specific. The greater the kyphosis or the smaller the lordosis of a spinal region, the more severe the degree of ossification in the corresponding region. LEVEL OF EVIDENCE: 3.

Sacroiliac joint degeneration in degenerative lumbar spondylolisthesis and related risk factors: a retrospective study.

OBJECTIVE: This study was performed to evaluate the degree of radiological sacroiliac joint (SIJ) degeneration in patients with degenerative lumbar spondylolisthesis (DLS). The related risk factors for SIJ degeneration were also investigated. METHODS: We retrospectively analyzed the lumbar and pelvic computed tomography (CT) scans of 303 patients with DLS admitted from January 2018 to December 2021. One hundred and fifty-six age-, gender-, and body mass index-matched patients without lumbar anomality who underwent lower abdominal or pelvic computed tomography scans were included in the control group. Sagittal parameters were measured on full-length lateral radiographs. Two protocols (Backlund's grade and Eno's classification) were used to assess SIJ degeneration. Univariate analysis and bivariate and multivariate regression analysis were performed to identify the factors affecting SIJ degeneration in patients with DLS. RESULTS: According to Backlund's grade and Eno's classification, SIJ degeneration was more severe in the DLS group than in the control group (P < 0.001). Multi-segment degenerative changes (P = 0.032), two-level DLS (P = 0.033), a history of hysterectomy (P < 0.001), lower extremity pain (P = 0.016), and pelvic pain (P = 0.013) were associated with more significant SIJ degeneration as assessed by Backlund's grade. The results of Pearson's correlation analysis showed positive correlation between the sagittal vertical axis and SIJ degeneration (r = 0.232, P = 0.009). The multivariate linear regression analysis showed that a history of hysterectomy was significantly correlated with SIJ degeneration in patients with DLS (r = 1.951, P = 0.008). CONCLUSIONS: SIJ degeneration was more severe in patients with than without DLS. We should take SIJ degeneration into consideration when diagnosing and treating DLS especially those who had undergone previous hysterectomy or showed sagittal malalignment.

Effects of cottonseed meal on performance, gossypol residue, liver function, lipid metabolism, and cecal microbiota in geese.

A total of 240 28-d-old male goslings were used to investigate the effects of cottonseed meal (CSM) on performance, gossypol residue, liver function, lipid metabolism, and cecal microbiota. All birds were randomly allotted into five groups (eight goslings/replicate, six replicates/group) and subjected to a 35-d experiment. Five isonitrogenous and isoenergetic diets were formulated to produce diets in which 0% (control), 25% (CSM25), 50% (CSM50), 75% (CSM75), and 100% (CSM100) of protein from soybean meal was replaced by protein from CSM. The free gossypol contents in the five diets were 0, 44, 92, 135, and 183 mg/kg, respectively. Dietary CSM did not affect the growth performance from 29 to 63 d and carcass traits at 63 d (P > 0.05). Liver gossypol residues were influenced (P < 0.05) by dietary CSM and increased linearly (P < 0.05) and quadratically (P < 0.05) as dietary CSM increased. The malondialdehyde content of the liver was lower in the CSM100 group than in the other groups (P < 0.05). Serum triglyceride and low-density lipoprotein cholesterol were influenced (P < 0.05) by dietary CSM and increased linearly (P < 0.05) with increasing dietary CSM. Dietary CSM altered (P < 0.05) the composition of some fatty acids in the liver and breast muscle. The concentration of linolenic acid and Σn-3 polyunsaturated fatty acid (PUFA) in the liver and breast muscle decreased linearly, but the Σn-6/Σn-3 PUFA ratio increased linearly with increasing dietary CSM (P < 0.05). Dietary CSM affected (P < 0.05) the hepatic gene expression of fatty acid synthase (FAS), acetyl-CoA carboxylase (ACC), and apolipoprotein B (ApoB). As the dietary CSM concentration increased, the hepatic gene expression of FAS increased linearly (P < 0.05) and quadratically (P < 0.05), but the hepatic gene expression of ACC and ApoB increased linearly (P < 0.05). The CSM diet decreased the relative abundance of the Bacteroidota and Bacteroides (P < 0.05), and the CSM50 diet increased the relative abundance of the Firmicutes and Colidextribacter (P < 0.05) compared to the control group. Overall, these results show that dietary CSM has no adverse effects on the performance of goslings from 29 to 63 d. However, CSM affected organismal lipid metabolism, reduced products' edible value, and adaptively altered cecum microbiota.

The shortage of feed resources and the rising price have become one of the significant challenges for animal husbandry worldwide. Considering the strong tolerance and adaptability to roughage of geese, less expensive crop byproducts are used in goose feed by animal nutritionists. Cottonseed meal (CSM) is a potential substitute for soybean meal, and the main concern for its use in poultry feed is free gossypol. This study aimed to investigate the effects of CSM on the performance, gossypol residue, liver function, lipid metabolism, and cecal microbiota in geese. Results showed that dietary CSM has no adverse effects on the performance and liver function of goslings. However, gossypol residue in goose liver increased with increasing dietary CSM. Besides, CSM affected organismal lipid metabolism, altered the tissue fatty acid composition, and adaptively changed cecum microbial microbiota. In summary, CSM is a good dietary protein source for geese, but further attention may be needed to its use for the edible value of goose products.

The upregulation of NLRP3 inflammasome in dorsal root ganglion by ten-eleven translocation methylcytosine dioxygenase 2 (TET2) contributed to diabetic neuropathic pain in mice.

BACKGROUND: The nucleotide oligomerization domain (NOD)-like receptor family pyrin domain containing 3 (NLRP3) in dorsal root ganglion (DRG) contributes to pain hypersensitivity in multiple neuropathic pain models, but the function of the NLRP3 in diabetic neuropathic pain (DNP) and the regulation mechanism are still largely unknown. Epigenetic regulation plays a vital role in the controlling of gene expression. Ten-eleven translocation methylcytosine dioxygenase 2 (TET2) is a DNA demethylase that contributes to transcriptional activation. TET2 is also involved in high glucose (HG)-induced pathology. METHODS: DNP was induced in mice via the intraperitoneal injection of streptozotocin (STZ) for five consecutive days and the mechanical threshold was evaluated in STZ-diabetic mice by using von Frey hairs. The expression level of the NLRP3 pathway and TET2 in DRG were determined through molecular biology experiments. The regulation of the NLRP3 pathway by TET2 was examined in in vitro and in vivo conditions. RESULTS: In the present research, we first established the DNP model and found that NLRP3 pathway was activated in DRG. The treatment of NLRP3 inhibitor MCC950 alleviated the mechanical allodynia of DNP mice. Then we revealed that in STZ-diabetic mice DRG, the genomic DNA was demethylated, and the expression of DNA demethylase TET2 was increased evidently. Using RNA-sequencing analysis, we found that the expression of Txnip, a gene that encodes a thioredoxin-interacting protein (TXNIP) which mediates NLRP3 activation, was elevated in the DRG after STZ treatment. In addition, knocking down of TET2 expression in DRG using TET2-siRNA suppressed the mRNA expression of Txnip and subsequently inhibited the expression/activation of NLRP3 inflammasome in vitro and in vivo as well as relieved the pain sensitivity of DNP animals. CONCLUSION: The results suggested that the upregulation of the TXNIP/NLRP3 pathway by TET2 in DRG was involved in the pain hypersensitivity of the DNP model.

Inhibition of the LRRC8A channel promotes microglia/macrophage phagocytosis and improves outcomes after intracerebral hemorrhagic stroke.

Promoting microglial/macrophage (M/Mφ) phagocytosis accelerates hematoma clearance and improves the prognosis of intracerebral hemorrhagic stroke (ICH). Cation channels such as Piezo1 modulate bacterial clearance by regulating M/Mφ. Whether LRRC8A, an anion channel, affects M/Mφ erythrophagocytosis and functional recovery after ICH was investigated here. We found that LRRC8A is highly expressed on M/Mφ in the perihematomal region of ICH mice. Conditional knockout of Lrrc8a in M/Mφ or treatment with an LRRC8A channel blocker accelerated hematoma clearance, reduced neuronal death, and improved functional recovery after ICH. Mechanistically, the LRRC8A channel inhibition promoted M/Mφ phagocytosis by activating AMP-activated protein kinase (AMPK), thereby inducing nuclear translocation of nuclear factor-erythroid 2 related factor 2 (Nrf2) and increasing Cd36 transcription. Our findings illuminate the regulation of M/Mφ phagocytosis by the LRRC8A channel via the AMPK-Nrf2-CD36 pathway after ICH, suggesting that LRRC8A is a potential target for hematoma clearance in ICH treatment.

The contribution of spinal dorsal horn astrocytes in neuropathic pain at the early stage of EAE.

Reactive astrocytes play a complex role in multiple sclerosis, and the astrocytes reactivity is an important factor in the pathogenesis of pain. It is of great significance to explore the genesis and development mechanism of pain in the early stage of multiple sclerosis (MS) for early intervention of the disease. This study aims to explore astrocyte reactivity at different stages of the experimental autoimmune encephalomyelitis (EAE) model, a mouse model of MS, and the role of astrocytes in the pain in the early stage of the EAE. In this study, we demonstrated that spinal dorsal horn astrocytes were activated in the pre-clinical stage of EAE mice, and the inhibition of spinal cord astrocyte reactivity effectively alleviates pain symptoms in EAE mice. On the other hand, spinal cord microglia were not directly participated in the early EAE pain. Moreover, the ion channel LRRC8A mediated the reactivity of spinal dorsal horn astrocytes by regulating the STAT3 pathway, therefore playing a role in the early pain of EAE.