Coordinated changes in mRNA turnover, translation, and RNA processing bodies in bronchial epithelial cells following inflammatory stimulation.

Bronchial epithelial cells play a pivotal role in airway inflammation, but little is known about posttranscriptional regulation of mediator gene expression during the inflammatory response in these cells. Here, we show that activation of human bronchial epithelial BEAS-2B cells by proinflammatory cytokines interleukin-4 (IL-4) and tumor necrosis factor alpha (TNF-alpha) leads to an increase in the mRNA stability of the key chemokines monocyte chemotactic protein 1 and IL-8, an elevation of the global translation rate, an increase in the levels of several proteins critical for translation, and a reduction of microRNA-mediated translational repression. Moreover, using the BEAS-2B cell system and a mouse model, we found that RNA processing bodies (P bodies), cytoplasmic domains linked to storage and/or degradation of translationally silenced mRNAs, are significantly reduced in activated bronchial epithelial cells, suggesting a physiological role for P bodies in airway inflammation. Our study reveals an orchestrated change among posttranscriptional mechanisms, which help sustain high levels of inflammatory mediator production in bronchial epithelium during the pathogenesis of inflammatory airway diseases

Analysis of cognitive function and its related factors after treatment in Meniere’s disease

A growing body of research recently suggested the association between vestibular dysfunction and cognitive impairment. Meniere’s disease (MD), a common clinical vestibular disorder, is usually accompanied by hearing loss and emotional stress, both of which may mediate the relationship between vestibule dysfunction and cognition. It is currently unknown whether the cognitive decline in MD patients could improve through treatment and how it relates to multiple clinical characteristics, particularly the severity of vertigo. Therefore, in the present study, the MD patients were followed up for 3, 6, and 12 months after treatment, and the cognitive functions, vertigo symptoms, and related physical, functional, and emotional effects of the patients were assessed using the Montreal Cognitive Assessment (MoCA) and Dizziness Handicap Inventory (DHI), aiming to explore the change in cognition before and after therapy and the correlation with various clinical features. It was found that cognitive decline in MD patients compared to healthy controls before therapy. Importantly, this cognitive impairment could improve after effective therapy, which was related to the severity of vertigo, especially in functional and physical impacts. Our results support the view that vestibular dysfunction is a potentially modifiable risk factor for cognitive decline

Multi-Hop Question Generation with Knowledge Graph-Enhanced Language Model

The task of multi-hop question generation (QG) seeks to generate questions that require a complex reasoning process that spans multiple sentences and answers. Beyond the conventional challenges of what to ask and how to ask, multi-hop QG necessitates sophisticated reasoning from dispersed evidence across multiple sentences. To address these challenges, a knowledge graph-enhanced language model (KGEL) has been developed to imitate human reasoning for multi-hop questions.The initial step in KGEL involves encoding the input sentence with a pre-trained GPT-2 language model to obtain a comprehensive semantic context representation. Next, a knowledge graph is constructed using the entities identified within the context. The critical information in the graph that is related to the answer is then utilized to update the context representations through an answer-aware graph attention network (GAT). Finally, the multi-head attention generation module (MHAG) is performed over the updated latent representations of the context to generate coherent questions. Human evaluations demonstrate that KGEL generates more logical and fluent multi-hop questions compared to GPT-2. Furthermore, KGEL outperforms five prominent baselines in automatic evaluations, with a BLEU-4 score that is 27% higher than that of GPT-2

Including ramie (Boehmeria nivea L. Gaud) in the diet of dairy cows: effects on production performance, milk composition, rumen fermentation, and nutrient digestion

This experiment was conducted to examine the effects of substituting mixed silage (fresh ramie: dry rice straw = 80: 20; kg: kg) composed of fresh forage ramie, also known as ‘China grass’, Boehmeria nivea L. Gaud (a nettle native to Asia) and rice straw for corn silage and alfalfa hay on the production performance, milk components, rumen fermentation parameters, and nutrient digestion in dairy cows. Thirty multiparous Chinese Holstein cows (629 ± 59.2 kg of BW, 25 ± 4.7 kg of milk yield, and 100 ± 18 DIM; mean ± SD) were randomly divided into three groups. The experimental treatments containing three diets, each consisting of differing proportions of mixed silage (0%, 10%, and 20%, designated as CON, MS1, and MS2, respectively) as a substitution for corn silage and alfalfa hay. There were no effects of mixed silage diets on dry matter intake (DMI), 3.5% fat-corrected milk (FCM) yield, milk protein percentage, milk somatic cell count, and milk urea nitrogen (MUN), but linearly increased total solids (p = .03) and milk fat percentage (p = .001) in cows fed the MS1 and MS2 diets. Feeding mixed silage diets linearly reduced milk yield (p = .01) and milk lactose percentage (p = .01), and had linearly increased rumen pH values (p = .01). There were no observable differences in other rumen fermentation parameters between the mixed silage and control diets. Mixed silage diets exhibited increased DM digestibility (linearly, p = .04; quadratically, p = .017) and linearly decreased crude protein digestibility (p = .05), but we observed no differences in the apparent total-tract digestibility of EE, NDF, and ADF between the control and mixed silage diets. In conclusion, using ramie and rice straw mixed silage proved beneficial by increasing milk fat percentage, milk solids, and DM digestibility, ramie could be used as a potential forage resource in dairy cow diets.Highlights We tested the effects of substituting alfalfa hay and corn silage with mixed silage composed of fresh forage ramie and rice straw on dairy cows Mixed silage diets had no effect on dry matter intake (DMI) Mixed silage diets reduced milk yield and milk lactose percentag

Methylglyoxal reduces cell viability and mitochondrion membrane potential of human brain microvascular endothelial cell line hCMEC/D3

Objective To explore the effects and mechanism of methylglyoxal (MGO) in human brain microvascular endothelial cell line (hCMEC/D3). Methods The hCMEC/D3 cells were incubated with MGO(0.25, 0.50, 0.75, 1.00 and 1.25 mmol/L) for 24 h. Cell viability was detected by CCK-8 assay. Cellular LDH and SOD activities were determined by commercially available kits. The change of mitochondrion membrane potential was assessed by JC-1 probe. MitoSOX probe was used to observe mitochondrial reactive oxygen species (mROS) release. Results Compared with control, cell viability of hCMEC/D3 treated with MGO decreased in a concentration-dependent manner (P＜0.01). Increased LDH activity and decreased SOD activity were observed with MGO exposure(P＜0.01). The decrease of mitochondrion membrane potential and the excessive increase of mROS were induced after MGO exposure for 12 h, which were detected by JC-1 or MitoSOX probe. Conclusions MGO reduces cell viability and mitochondria membrane potential in hCMEC/D3 cells with excessive mROS generation

Genetic diversity and population structure of the endangered Japanese sea cucumber (Apostichopus japonicus) in natural seas of northern China

Overfishing and habitat destruction have decimated wild sea cucumber (Apostichopus japonicus) stocks, causing this species to become endangered. Several conservation measures have been implemented to assist in the recovery of A. japonicus. Due to the low efficiency of conventional domestication methods and the current prevalence of bottom-sowing culture in marine ranches, it is urgent to evaluate the genetic diversity and population structure of sea cucumber geographical groups in the natural seas of northern China using genome-wide molecular markers. In this study, six sea cucumber geographical groups were collected from marine ranches in Shandong, Hebei, and Liaoning provinces. A total of 18,191 high-quality genome-wide single nucleotide polymorphisms (SNPs) were identified in 70 A. japonicus individuals using 2b-RAD technology. These SNPs were used to assess the genetic diversity and population structure of the six groups. Compared with previous studies, these six geographical groups showed much lower levels of genetic diversity both among and between groups, with all individuals clustering into one population. These findings demonstrate the necessity of continuous genetic monitoring of A. japonicus in the natural seas of northern China. Furthermore, this study serves as a valuable reference for future genome-assisted breeding and germplasm improvement in this economically critical species

Overexpression of Cell Surface Cytokeratin 8 in Multidrug-Resistant MCF-7/MX Cells Enhances Cell Adhesion to the Extracellular Matrix1

Accumulating evidence suggests that multiple complex mechanisms may be involved, simultaneously or complementarily, in the emergence and development of multidrug resistance (MDR) in various cancers. Cell adhesion-mediated MDR is one such mechanism. In the present study, we initially observed increased cell adhesion to extracellular matrix proteins by the MDR human breast tumor cell line MCF-7/MX compared to its parental cells. We then used a strategy that combined antibody-based screening technique and mass spectrometry-based proteomics to identify membrane proteins that contribute to the enhanced adhesion of MCF-7/MX cells. Using MCF-7/MX cells as immunogen, we isolated a mouse monoclonal antibody, 9C6, that preferentially reacts with MCF-7/MX cells over the parental MCF-7 cells. The molecular target of 9C6 was identified as cytokeratin 8 (CK8), which was found to be overexpressed on the cell surface of MCF-7/MX cells. We further observed that down-regulation of cell surface levels of CK8 through siRNA transfection significantly inhibited MCF-7/MX cell adhesion to fibronectin and vitronectin. In addition, anti-CK8 siRNA partially reversed the MDR phenotype of MCF-7/MX cells. Taken together, our results suggest that alterations in the expression level and cellular localization of CK8 may play a significant role in enhancing the cellular adhesion of MDR MCF-7/MX cells

Versatile applications of transcriptional pulsing to study mRNA turnover in mammalian cells

Development of transcriptional pulsing approaches using the c-fos and Tet-off promoter systems greatly facilitated studies of mRNA turnover in mammalian cells. However, optimal protocols for these approaches vary for different cell types and/or physiological conditions, limiting their widespread application. In this study, we have further optimized transcriptional pulsing systems for different cell lines and developed new protocols to facilitate investigation of various aspects of mRNA turnover. We apply the Tet-off transcriptional pulsing strategy to investigate ARE-mediated mRNA decay in human erythroleukemic K562 cells arrested at various phases of the cell cycle by pharmacological inhibitors. This application facilitates studies of the role of mRNA stability in control of cell-cycle dependent gene expression. To advance the investigation of factors involved in mRNA turnover and its regulation, we have also incorporated recently developed transfection and siRNA reagents into the transcriptional pulsing approach. Using these protocols, siRNA and DNA plasmids can be effectively cotransfected into mouse NIH3T3 cells to obtain high knockdown efficiency. Moreover, we have established a tTA-harboring stable line using human bronchial epithelial BEAS-2B cells and applied the transcriptional pulsing approach to monitor mRNA deadenylation and decay kinetics in this cell system. This broadens the application of the transcriptional pulsing system to investigate the regulation of mRNA turnover related to allergic inflammation. Critical factors that need to be considered when employing these approaches are characterized and discussed