Dedifferentiation-like reprogramming of degenerative nucleus pulposus cells into notochordal-like cells by defined factors.

The extensive degeneration of functional somatic cells and the depletion of endogenous stem/progenitor populations present significant challenges to tissue regeneration in degenerative diseases. Currently, a cellular reprogramming approach enabling directly generating corresponding progenitor populations from degenerative somatic cells remains elusive. The present study focused on intervertebral disc degeneration (IVDD) and identified a three-factor combination (OCT4, FOXA2, TBXT [OFT]) that could induce the dedifferentiation-like reprogramming of degenerative nucleus pulposus cells (dNPCs) toward induced notochordal-like cells (iNCs). Single-cell transcriptomics dissected the transitions of cell identity during reprogramming. Further, OCT4 was found to directly interact with bromodomain PHD-finger transcription factor to remodel the chromatin during the early phases, which was crucial for initiating this dedifferentiation-like reprogramming. In rat models, intradiscal injection of adeno-associated virus carrying OFT generated iNCs from in situ dNPCs and reversed IVDD. These results collectively present a proof-of-concept for dedifferentiation-like reprogramming of degenerated somatic cells into corresponding progenitors through the development of a factor-based strategy, providing a promising approach for regeneration in degenerative disc diseases.

Associations between pre-infection serum vitamin D concentrations and Omicron COVID-19 incidence, severity and reoccurrence in elderly individuals.

OBJECTIVE: Previous studies suggest a link between vitamin D status and COVID-19 susceptibility in hospitalised patients. This study aimed to investigate whether vitamin D concentrations in elderly individuals were associated with their susceptibility to Omicron COVID-19 incidence, the severity of the disease and the likelihood of reoccurrence during the era of the post-'zero-COVID-19' policies in China. DESIGN: In this retrospective study, participants were categorised into three groups based on their 25(OH)D concentrations: deficiency (< 20 ng/ml), insufficiency (20 to < 30 ng/ml) and sufficiency (≥ 30 ng/ml). The demographic and clinical characteristics, comorbidities and the incidence rate, reoccurrence rate and severity of Omicron COVID-19 were retrospectively recorded and analysed by using hospital information system data and an online questionnaire survey. SETTING: China. PARTICIPANTS: 222 participants aged 60 years or older from a health management centre. RESULTS: Our findings revealed significant differences in the incidence (P = 0·03) and recurrent rate (P = 0·02) of Omicron COVID-19 among the three groups. Participants with lower 25(OH)D concentrations (< 20 ng/ml) exhibited higher rates of initial incidence and reoccurrence and a greater percentage of severe and critical cases. Conversely, individuals with 25(OH)D concentrations ≥ 30 ng/ml had a higher percentage of mild cases (P = 0·003). Binary and ordinal logistic regression models indicated that vitamin D supplementation was not a significant risk factor for COVID-19 outcomes. CONCLUSIONS: In the elderly population, pre-infection vitamin D deficiency was associated with increased susceptibility to incidence, severity of illness and reoccurrence rates of Omicron COVID-19.

Factors Influencing Intentions of People with Hearing Impairments to Use Augmented Reality Glasses as Hearing Aids.

The advent and progression of AR (augmented reality) technology, coupled with the emergence of AR hearing aid glasses, offer a novel opportunity for people with hearing impairments (PHI). This study aims to explore the intention of this population to employ AR hearing aid glasses as their choice of hearing aid device and the specific factors influencing their preference. This study utilized the partial least squares SEM (PLS-SEM) analytical method to create structural equation model for intentions of PHI to use AR glasses as hearing aids. Data were gathered from on-site experiences across multiple locations; a total of 189 valid questionnaires from individuals with varying degrees of hearing disabilities were used for statistical analysis. According to the data analysis results, we discovered that functionality quality, perceived interaction speed, and perceived usability significantly influence communication effectiveness. Further, communication effectiveness positively influences confidence and societal perception, and the latter has a positive impact on information. Both of these factors positively influence behavioral intention. Based on these findings, this study offers design recommendations for AR hearing aid glasses to cater to the specific needs of PHI, aiming to enhance their quality of life. Furthermore, this study provides pivotal insights for the prospective growth of this emerging industry.

Non-POEMS osteosclerotic multiple myeloma: Clinical characteristics and differential diagnosis.

Osteosclerosis in multiple myeloma (MM) is typically associated with rare POEMS syndrome, characterized by polyneuropathy (P), organomegaly (O), endocrinopathy (E), M-protein (M), and skin changes (S). However, osteosclerosis in multiple myeloma (MM) without POEMS syndrome, defined as non-POEMS Osteosclerotic MM, is exceedingly rare. We report a 70-year-old man with rib pain, remarkably high bone mineral density and diffuse osteosclerosis. The diagnosis of non-POEMS osteosclerotic MM was confirmed by biopsy and aspiration of bone marrow through surgery. A literature review spanning from 1990 identified 12 cases of similar non-POEMS osteosclerotic MM, including 5 males and 7 females with a mean age of 59.7 ± 10.6 years. The non-POEMS osteosclerotic MM can be divided into two subtypes, the osteosclerotic lesion subtype and the diffuse osteosclerosis subtype. Absence of polyneuropathy and organomegaly are the main factors that differentiate non-POEMS osteosclerotic MM from POEMS. A hyperactive osteoblastic process might be the etiology of diffuse osteosclerosis. Further research is needed to understand its etiology and pathophysiology.

Is there any way to increase consumers' purchase intention regarding surplus food blind-boxes? An exploratory study.

As an emerging business model, merchants are selling surplus food in blind boxes, which provides new opportunities for reducing waste and promoting sustainable development. Surplus food blind boxes, however, have not been extensively researched in terms of design and marketing strategies. Therefore, we are examining how to increase consumers' purchase intention regarding surplus food blind boxes as well as the relationship between purchase intention, continuous intention, and recommendation intention. In this paper, two studies are conducted. As part of study 1, We used factor analysis to explore the reasons that influence consumers to purchase. In study 2, we utilized structural equation modeling to examine the impact of purchase intention on continuous intention and recommendation intention. Results indicate that food quality, perceived sustainability, and conspicuous consumption are the most important positive factors. The most important negative factors are perceived food risk, resistance to sales techniques, taste anxiety, and inadequate marketing strategies. Based on our findings, we constructed a path relationship between purchase intention, continuous intention, and recommendation intention. The purchase intention of the surplus food blind box users directly influences the recommendation intention and the continuous intention, while the recommendation intention indirectly influences the continuous intention. This study did not demonstrate that gender had a moderating effect. Meanwhile, it provides a reference for the actual marketing management of surplus food blind box merchants.

Transcriptome Analysis of Differentially Expressed Genes and Molecular Pathways Involved in C2C12 Cells Myogenic Differentiation.

Muscles are essential tissues responsible for movement, stability, and metabolism, playing a crucial role in human health and well-being. A comprehensive understanding of muscle differentiation processes is imperative for combating muscle degenerative diseases such as muscular dystrophy. In this study, C2C12 cells were induced to differentiate into myotubes in vitro. Phenotypic changes were observed utilizing Gimsa and immunofluorescent staining techniques. RNA sequencing was conducted at distinct time points (0, 2, 4, and 7 days) during the differentiation process. To elucidate the underlying molecular mechanisms, differential expression analysis, gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, and Gene Set Enrichment Analysis (GSEA) were performed. Soft clustering of time series gene expression was employed to establish the expression patterns of differentially expressed genes (DEGs) at various time points during myogenesis. Additionally, quantitative reverse transcription PCR was utilized to validate gene expression from RNA-seq data at the mRNA level. Throughout the myogenic differentiation of C2C12 cells, notable morphological changes were observed, with myoblasts forming multinucleated myotubes by day 4 and plump elongated structures by day 7. Gene expression analysis revealed a substantial increase in DEGs as differentiation progressed, with a significant rise in DEGs from day 0 to day 7. Enrichment analysis highlighted key biological processes and pathways involved, including signal transduction and immune system processes, as well as pathways like chemokine and calcium signaling. Noise-robust soft clustering identified distinct temporal gene expression patterns, categorizing genes into upregulated, downregulated, and biphasic response clusters. The MYH family exhibited diverse expression changes, with Myh3, Myh13, Myh6, Myh7, Myh2, Myh8, Myh14, Myh7b, Myh1, and Myh4 upregulated, Myh10, Myh9, and Myh12 downregulated. Key transcription factors displayed dynamic expression patterns, which was crucial for the regulation of myoblast differentiation. A comprehensive and dynamic transcriptomic analysis of the C2C12 myoblast differentiation process has significantly enhanced our understanding of the key genes and biological pathways involved in myogenesis.

A novel rat model of annulus fibrosus injury for intervertebral disc degeneration.

BACKGROUND CONTEXT: In clinical practice, acute trauma and chronic degeneration of the annulus fibrosus (AF) can promote further degeneration of the intervertebral disc (IVD). Therefore, it is critical to understand the AF repair process and its consequences on IVD. However, the lack of cost-effective and reproducible in vivo animal models of AF injury has limited research development in this field. PURPOSES: The purpose of this study was to establish and evaluate the utility of a novel animal model for full-thickness AF injury. Three foci were proposed: (1) whether this new modeling method can cause full-layer AF damage; (2) the repair processes and pathological changes in the damaged area after AF injury, and (3) the morphological and histological changes in the IVD are after AF injury. STUDY DESIGN/SETTING: In vivo rat AF injury model with characterization of AF damage repair, IVD degeneration. METHODS: A total of 72,300 g male rats were randomly assigned to one of the two groups: experimental or sham. Annulus fibrosus was separated layer by layer under the microscope with a #11 blade up to the AF- nucleus pulpous (NP) junction. The repair process of the horizontal AF and morphological changes in the sagittal IVD were evaluated with HE staining. Sirius red staining under polarized light. Immunofluorescence was conducted to analyze changes in the expression of COL1 and COL3 in the AF injury area and 8-OHdg, IL-6, MMP13, FSP1, and ACAN in the IVD. The disc height and structural changes after AF injury were measured using X-ray and contrast-enhanced micro-CT. Additionally, the resistance of the AF to stretching was analyzed using three-point bending. RESULTS: Annulus fibrosus-nucleus pulpous border was identified to stably induce the full-thickness AF injury without causing immediate NP injury. The AF repair process after injury was slow and expressed inflammation factors continuously, with abundant amounts of type III collagen appearing in the inner part of the AF. The scar at the AF lesion had decreased resistance to small molecule penetration and weakened tensile strength. Full-thickness AF injury induced disc degeneration with loss of disc height, progressive unilateral vertebral collapse, and ossification of the subchondral bone. Inflammatory-induced degeneration and extracellular matrix catabolism gradually appeared in the NP and cartilage endplate (CEP). CONCLUSIONS: We established a low-cost and reproducible small animal model of AF injury which accurately replicated the pathological state of the limited AF self-repair ability and demonstrated that injury to the AF alone could cause further degeneration of the IVD. CLINICAL RELEVANCE: This in vivo rat model can be used to study the repair process of the AF defect and pathological changes in the gradual degeneration of IVD after AF damage. In addition, the model provides an experimental platform for in vivo experimental research of potential clinical therapeutics.

Silencing circATXN1 in Aging Nucleus Pulposus Cell Alleviates Intervertebral Disc Degeneration via Correcting Progerin Mislocalization.

Circular RNAs (circRNAs) play a critical regulatory role in degenerative diseases; however, their functions and therapeutic applications in intervertebral disc degeneration (IVDD) have not been explored. Here, we identified that a novel circATXN1 highly accumulates in aging nucleus pulposus cells (NPCs) accountable for IVDD. CircATXN1 accelerates cellular senescence, disrupts extracellular matrix organization, and inhibits mitochondrial respiration. Mechanistically, circATXN1, regulated by heterogeneous nuclear ribonucleoprotein A2B1-mediated splicing circularization, promotes progerin translocation from the cell nucleus to the cytoplasm and inhibits the expression of insulin-like growth factor 1 receptor (IGF-1R). To demonstrate the therapeutic potential of circATXN1, siRNA targeting the backsplice junction of circATNX1 was screened and delivered by tetrahedral framework nucleic acids (tFNAs) due to their unique compositional and tetrahedral structural features. Our siRNA delivery system demonstrates superior abilities to transfect aging cells, clear intracellular ROS, and enhanced biological safety. Using siRNA-tFNAs to silence circATXN1, aging NPCs exhibit reduced mislocalization of progerin in the cytoplasm and up-regulation of IGF-1R, thereby demonstrating a rejuvenated cellular phenotype and improved mitochondrial function. In vivo, administering an aging cell-adapted siRNA nucleic acid framework delivery system to progerin pathologically expressed premature aging mice (zmpste24-/-) can ameliorate the cellular matrix in the nucleus pulposus tissue, effectively delaying IVDD. This study not only identified circATXN1 functioning as a cell senescence promoter in IVDD for the first time, but also successfully demonstrated its therapeutic potential via a tFNA-based siRNA delivery strategy.

Research on high school students' behavior in art course within a virtual learning environment based on SVVR.

Introduction: Students who use spherical video-based virtual reality (SVVR) teaching materials for learning are able to gain more self-regulated, explorative, and immersive experiences in a virtual environment. Using SVVR teaching materials in art courses can present diverse and unique teaching effects, while also leading to the emergence of students' flow states. Methods: Therefore, through an art course teaching experiment, this study investigated 380 high school students and used structural equation modeling to analyze the antecedents and outcomes of students' flow state in using SVVR teaching materials. Results: The results show that in using SVVR teaching materials in art courses, more attention should be paid to the control and telepresence in the antecedents of students' flow state. Discussion: Only when they obtain better flow experiences can they have higher perceived usefulness and satisfaction with the content of the art course, as well as stronger intentions to continue using it. These results can provide a reference for the development and use of SVVR teaching materials in high school art courses.

Correction: The impact of reusable tableware packaging combined with environmental propaganda on consumer behaviour in online retail.

[This corrects the article DOI: 10.1371/journal.pone.0264562.].

A study on how to improve users' perceived playfulness in and continuance intention with VR technology to paint in virtual natural landscapes.

COVID-19 has adversely affected public access to public green spaces. As a means of interacting with nature, parks and green spaces are an important aspect of residents' daily lives. In this study, the focus is on new digital solutions, such as the experience of painting in virtual natural settings through virtual reality technologies. This study examines factors that affect the user's perceived playfulness and continuance intention to paint in a virtual environment. A total of 732 valid samples were collected through a questionnaire survey, and a theoretical model was developed through structural equation model by analyzing attitude, perceived behavioral control, behavioral intention, continuance intention, and perceived playfulness. Results demonstrate that perceived novelty and perceived sustainability increase the positive attitude of users toward VR painting functions, whereas perceived interactivity and aesthetics have no impact on it within VR painting context. As users use VR painting, they are more concerned about time and money than equipment compatibility. This makes resource facilitating conditions a more influential factor for perceived behavior control than technology facilitating conditions.

Metabolic Glycoengineering: A Promising Strategy to Remodel Microenvironments for Regenerative Therapy.

Cell-based regenerative therapy utilizes the differentiation potential of stem cells to rejuvenate tissues. But the dynamic fate of stem cells is calling for precise control to optimize their therapeutic efficiency. Stem cell fate is regulated by specific conditions called "microenvironments." Among the various factors in the microenvironment, the cell-surface glycan acts as a mediator of cell-matrix and cell-cell interactions and manipulates the behavior of cells. Herein, metabolic glycoengineering (MGE) is an easy but powerful technology for remodeling the structure of glycan. By presenting unnatural glycans on the surface, MGE provides us an opportunity to reshape the microenvironment and evoke desired cellular responses. In this review, we firstly focused on the determining role of glycans on cellular activity; then, we introduced how MGE influences glycosylation and subsequently affects cell fate; at last, we outlined the application of MGE in regenerative therapy, especially in the musculoskeletal system, and the future direction of MGE is discussed.

Controlled extracellular vesicles release from aminoguanidine nanoparticle-loaded polylysine hydrogel for synergistic treatment of spinal cord injury.

Pharmaceutical treatments are critical for the acute and subacute phases of spinal cord injury (SCI) and significantly impact patients' prognoses. However, there is a lack of a precise, multitemporal, integrated drug delivery system for medications administered in both phases. In this study, we prepare a hybrid polylysine-based hydrogel (PBHEVs@AGN) comprising short-term release of pH-responsive aminoguanidine nanoparticles (AGN) and sustained release of extracellular vesicles (EVs) for synergistic SCI treatment. When AGN is exposed to the acidic environment at the injury site, it quickly diffuses out of the hydrogel and releases the majority of the aminoguanidine within 24 h, reducing oxidative stress in lesion tissues. Enriched EVs are gradually released from the hydrogel and remain in the tissue for weeks, providing a long-term anti-inflammatory effect and further ensuring axonal regeneration. Fast-releasing aminoguanidine can cooperate with slow-release EVs to treat SCI more effectively by reducing the production of proinflammatory cytokines and blocking the TLR4/Myd88/NF-κB inflammatory pathway, creating a sustained anti-inflammatory microenvironment for SCI recovery. Our in vivo experiments demonstrate that PBHEVs@AGN reduces the occurrence of scar tissue, encourages remyelination, and speeds up axonal regeneration. Herein, this multi-drug delivery system, which combines the acute release of aminoguanidine and the sustained release of EVs is highly effective for synergistically managing the challenging pathological processes after SCI.

Thermosensitive hydrogel-based GPR124 delivery strategy for rebuilding blood-spinal cord barrier.

Spinal cord injury (SCI) causes blood-spinal cord barrier (BSCB) disruption, leading to secondary damage, such as hemorrhagic infiltration, inflammatory response, and neuronal cell death. It is of great significance to rebuild the BSCB at the early stage of SCI to alleviate the secondary injury for better prognosis. Yet, current research involved in the reconstruction of BSCB is insufficient. Accordingly, we provide a thermosensitive hydrogel-based G protein-coupled receptor 124 (GPR124) delivery strategy for rebuilding BSCB. Herein, we firstly found that the expression of GPR124 decreased post-SCI and demonstrated that treatment with recombinant GPR124 could partially alleviate the disruption of BSCB post-SCI by restoring tight junctions (TJs) and promoting migration and tube formation of endothelial cells. Interestingly, GPR124 could also boost the energy metabolism of endothelial cells. However, the absence of physicochemical stability restricted the wide usage of GPR124. Hence, we fabricated a thermosensitive heparin-poloxamer (HP) hydrogel that demonstrated sustained GPR124 production and maintained the bioactivity of GPR124 (HP@124) for rebuilding the BSCB and eventually enhancing functional motor recovery post-SCI. HP@124 hydrogel can encapsulate GPR124 at the lesion site by injection, providing prolonged release, preserving wounded tissues, and filling injured tissue cavities. Consequently, it induces synergistically efficient integrated regulation by blocking BSCB rupture, decreasing fibrotic scar formation, minimizing inflammatory response, boosting remyelination, and regenerating axons. Mechanistically, giving GPR124 activates energy metabolism via elevating the expression of phosphoenolpyruvate carboxykinase 2 (PCK2), and eventually restores the poor state of endothelial cells. This research demonstrated that early intervention by combining GPR124 with bioactive multifunctional hydrogel may have tremendous promise for restoring locomotor recovery in patients with central nervous system disorders, in addition to a translational approach for the medical therapy of SCI.

Collagen Niches Affect Direct Transcriptional Conversion toward Human Nucleus Pulposus Cells via Actomyosin Contractility.

Cellular niches play fundamental roles in regulating cellular behaviors. However, the effect of niches on direct converted cells remains unexplored. In the present study, the specific combination of transcription factors is first identified to directly acquire induced nucleus pulposus-like cells (iNPLCs). Next, tunable physical properties of collagen niches are fabricated based on various crosslinking degrees. Collagen niches significantly affect actomyosin cytoskeleton and then influence the maturation of iNPLCs. Using gain- and loss of function approaches, the appropriate physical states of collagen niches are found to significantly enhance the maturation of iNPLCs through actomyosin contractility. Moreover, in a rat model of degenerative disc diseases, iNPLCs with collagen niches are transplanted into the lesion to achieve significant improvements. As a result, overexpression of transcription factors in human dermal fibroblasts are efficiently converted into iNPLCs and the optimal collagen niches affect cellular cytoskeleton and then facilitate iNPLCs maturation toward human nucleus pulposus cells. These findings encourage more in-depth studies toward the interactions of niches and direct conversion, which would contribute to the development of direct conversion.

Swelling-Mediated Mechanical Stimulation Regulates Differentiation of Adipose-Derived Mesenchymal Stem Cells for Intervertebral Disc Repair Using Injectable UCST Microgels.

Mechanical stimulation is an effective approach for controlling stem cell differentiation in tissue engineering. However, its realization in in vivo tissue repair remains challenging since this type of stimulation can hardly be applied to injectable seeding systems. Here, it is presented that swelling of injectable microgels can be transformed to in situ mechanical stimulation via stretching the cells adhered on their surface. Poly(acrylamide-co-acrylic acid) microgels with the upper critical solution temperature property are fabricated using inverse emulsion polymerization and further coated with polydopamine to increase cell adhesion. Adipose-derived mesenchymal stem cells (ADSCs) adhered on the microgels can be omnidirectionally stretched along with the responsive swelling of the microgels, which upregulate TRPV4 and Piezo1 channel proteins and enhance nucleus pulposus (NP)-like differentiation of ADSCs. In vivo experiments reveal that the disc height and extracellular matrix content of NP are promoted after the implantation with the microgels. The findings indicate that swelling-induced mechanical stimulation has great potential for regulating stem cell differentiation during intervertebral disc repair.

What do users care about? Research on user behavior of mobile interactive video advertising.

Mobile interactive video advertising is a relatively new type of media. A combination of interactivity and narrative is used to tell the video story to the audience. This study studied in three steps on mobile interactive video advertising to determine the design basis that should be paid attention to. Firstly, the research framework is proposed through expert interview and qualitative coding of the interview text. Secondly, research samples are made through surveys and conference discussions. The samples were designed into 30 types containing various interactive narrative and involvement. Finally, the qualitative results are verified in a quantitative way. The results show that in order to improve the audience's purchase intention, more attention should be paid to interactive narrative, subjective video quality assessment, immersion, satisfaction, and product involvement.

Examining the Influence of Using First-Person View Drones as Auxiliary Devices in Matte Painting Courses on College Students' Continuous Learning Intention.

In terms of the teaching process of matte painting, it is essential for students to develop a sound understanding of the relationship between virtual and physical environments. In this study, first-person view (FPV) drones are applied to matte painting courses to evaluate the effectiveness of the teaching, and to propose more effective design suggestions for FPV drones that are more suitable for teaching. This provides students with a better learning environment using a digital education system. The results of the study indicate that the flow experience, learning interest, and continuous learning intention of students who use FPV drones in matte painting are significantly greater than those of students who only utilize traditional teaching methods. Furthermore, the technology incentive model (TIM) was developed in this study after being verified by the structural equation model. The results demonstrate that the second-order construct 'technology incentive' comprising perceived interactivity, perceived vividness, and novel experience positively influence students' learning interest and continuous learning intentions under the mediation of flow experience.

Effects of AR Picture Books on German Teaching in Universities.

In this paper, we discuss the teaching effects of augmented reality (AR) technology in German instruction. We conducted one prestudy and three formal studies on German learners in China's mainland and Taiwan region. In the formal studies, a total of 120 students participated in the survey, allowing us to compare the differences in interest in learning between AR picture books and traditional picture books. A total of 114 students took part in the survey, which enabled us to compare the contribution of AR picture books to teaching when students' satisfaction and German proficiency were different. To improve satisfaction, 514 students participated in the survey regarding the influence of the interactive narrative design effect and peer learning on satisfaction with using AR picture books. The results suggest that when learning German with AR picture books, satisfaction is the key construct that determines students' learning states.

A study on the effects of perceived playfulness and emotional support in interactive learning environments for German language acquisition-A role-playing library system case.

Introduction: This study examines the emotional support offered by the non-player characters (NPCs) in an interactive learning environment, as well as the effects of the perceived playfulness of the interactive system on German language learning. Method: We developed a role-playing library system to serve this purpose. 2,377 Chinese Internet users were surveyed using online questionnaire. Results: A theoretical model of emotion- driven learning (ELM) was proposed based on the analysis results of valid recovered data. Additionally, NPCs were found to be effective in improving learning outcomes through emotional support. Discussion: An interactive education system may be able to enhance the perceived playfulness of learning in order to enhance the learning experience.