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Physical activity shows a positive correlation with overall health, and vigorous intermittent lifestyle physical activity (VILPA) similarly offers advantages in reducing the risk of all-cause mortality. Might the short-time mechanical stimuli be discernible to cells, eliciting commensurate physiological responses? The study's objective was to investigate the cellular response to short-time mechanical stimuli. Human umbilical cord-derived mesenchymal stem cells (hUCMSCs), isolated and thoroughly characterized, were subjected to various stimuli, including activation and mechanical stretching, with Ca2+ influx assessed through alterations in fluorescence intensity. Further validation of these findings was confirmed through short hairpin RNA (shRNA) and inhibitors. In addition, a comprehensive examination of PIEZO1 alterations was conducted through quantitative real-time polymerase chain reaction (qRT-PCR) and western blot (WB) techniques. The results shown different frequencies of stretching stimulation and durations induced varying degrees of Ca2+ influx. The most substantial increase occurred within 2–3 minutes in the group subjected to 0.5 Hz stretching for 2 minutes (p < 0.05). Stretching at 0.5 Hz resulted in significant elevation in PIEZO1 mRNA expression at 15 minutes and 1 hour. Additionally, stretching cause a gradual rise in PIEZO1 protein levels, with a notable peak observed at 2 hours. In conclusion, cells primarily sense short-time mechanical stimuli through PIEZO1, predominantly mediated by regulated Ca2+ influx. This underscores PIEZO1's crucial role in cellular responsiveness to transient mechanical cues, advancing our understanding of mechanosensory mechanisms in cellular physiology.

Abstract Objectives The main objective of this study was to establish a mouse model of spinal ligament ossification to simulate the chronic spinal cord compression observed in patients with ossification of the posterior longitudinal ligament (OPLL). The study also aimed to examine the mice's neurobiological, radiological, and pathological changes. Methods In the previous study, a genetically modified mouse strain was created using Crispr‐Cas9 technology, namely, Enpp1flox/flox/EIIa‐Cre (C57/B6 background), to establish the OPLL model. Wild‐type (WT) mice without compression were used as controls. Functional deficits were evaluated through motor score assessment, inclined plate testing, and gait analysis. The extent of compression was determined using CT imaging. Hematoxylin and eosin staining, luxol fast blue staining, TUNEL assay, immunofluorescence staining, qPCR, and Western blotting were performed to evaluate levels of apoptosis, inflammation, vascularization, and demyelination in the study. Results The results demonstrated a gradual deterioration of compression in the Enpp1flox/flox/EIIa‐Cre mice group as they aged. The progression rate was more rapid between 12 and 20 weeks, followed by a gradual stabilization between 20 and 28 weeks. The scores for spinal cord function and strength, assessed using the Basso Mouse Scale and inclined plate test, showed a significant decline. Gait analysis revealed a noticeable reduction in fore and hind stride lengths, stride width, and toe spread. Chronic spinal cord compression resulted in neuronal damage and activated astrocytes and microglia in the gray matter and anterior horn. Progressive posterior cervical compression impeded blood supply, leading to inflammation and Fas‐mediated neuronal apoptosis. The activation of Bcl2 and Caspase 3 was associated with the development of progressive neurological deficits (p

Abstract The precise regulation of cellular behaviors within a confined, crowded intracellular environment is highly amenable in diagnostics and therapeutics. While synthetic circuitry system through a concatenated chemical reaction network has rarely been reported to mimic dynamic self‐assembly system. Herein, a catalytic self‐defined circuit (CSC) for the hierarchically concatenated assembly of DNA domino nanostructures is engineered. By incorporating pre‐sealed symmetrical fragments into the preying hairpin reactants, the CSC system allows the hierarchical DNA self‐assembly via a microRNA (miRNA)‐powered self‐sorting catalytic hybridization reaction. With minimal strand complexity, this self‐sustainable CSC system streamlined the circuit component and achieved localization‐intensified cascaded signal amplification. Profiting from the self‐adaptively concatenated hybridization reaction, a reliable and robust method has been achieved for discriminating carcinoma tissues from the corresponding para‐carcinoma tissues. The CSC‐sustained self‐assembly strategy provides a comprehensive and smart toolbox for organizing various hierarchical DNA nanostructures, which may facilitate more insights for clinical diagnosis and therapeutic assessment.

BackgroundThe relationship between hormonal fluctuations in the reproductive system and the occurrence of low back pain (LBP) has been widely observed. However, the causal impact of specific variables that may be indicative of hormonal and reproductive factors, such as age at menopause (ANM), age at menarche (AAM), length of menstrual cycle (LMC), age at first birth (AFB), age at last live birth (ALB) and age first had sexual intercourse (AFS) on low back pain remains unclear.MethodsThis study employed Bidirectional Mendelian randomization (MR) using publicly available summary statistics from Genome Wide Association Studies (GWAS) and FinnGen Consortium to investigate the causal links between hormonal and reproductive factors on LBP. Various MR methodologies, including inverse-variance weighted (IVW), MR-Egger regression, and weighted median, were utilized. Sensitivity analysis was conducted to ensure the robustness and validity of the findings. Subsequently, Multivariate Mendelian randomization (MVMR) was employed to assess the direct causal impact of reproductive and hormone factors on the risk of LBP.ResultsAfter implementing the Bonferroni correction and conducting rigorous quality control, the results from MR indicated a noteworthy association between a decreased risk of LBP and AAM (OR=0.784, 95% CI: 0.689-0.891; p=3.53E-04), AFB (OR=0.558, 95% CI: 0.436-0.715; p=8.97E-06), ALB (OR=0.396, 95% CI: 0.226-0.692; p=0.002), and AFS (OR=0.602, 95% CI: 0.518-0.700; p=3.47E-10). Moreover, in the reverse MR analysis, we observed no significant causal effects of LBP on ANM, AAM, LMC and AFS. MVMR analysis demonstrated the continued significance of the causal effect of AFB on LBP after adjusting for BMI.ConclusionOur study explored the causal relationship between ANM, AAM, LMC, AFB, AFS, ALB and the prevalence of LBP. We found that early menarche, early age at first birth, early age at last live birth and early age first had sexual intercourse may decrease the risk of LBP. These insights enhance our understanding of LBP risk factors, offering valuable guidance for screening, prevention, and treatment strategies for at-risk women.

Background: Heterotopic ossification of tendons and ligaments (HOTL) is a common clinical condition characterized by the absence of discernible features and a lack of effective treatment. In vitro experiments have demonstrated that mechanical stimulation can induce cell differentiation toward osteogenesis, thereby promoting heterotopic ossification. Currently, there are few experimental designs aimed at inducing ligament stretching in mice, and the mechanism of heterotopic ossification may not entirely mirror that observed in clinical cases. Therefore, there is an urgent imperative to develop a novel and feasible animal model.Methods: In this study, all the Enpp1 gene deficiency mice (a mouse model with heterotopic ossification of multiple ligaments) were divided into three groups: the control group, the spinal brake group, and the hyperactive group (treadmill training group). An external spinal fixation device was designed to restrict mice’s spinal flexion and extension at 6 weeks of age. The brace was adjusted weekly according to the changes in the size of the mice. Additionally, treadmill training was used to increase activity in the spinal ligaments and Achilles tendons of the mice. Micro-CT scanning and HE staining were performed at 12, 20, and 28 W to evaluate the degree of ossification in the spinal ligament and Achilles tendon. What’s more, As one of the mechanical stimulation transduction signals, YAP plays a crucial role in promoting osteogenic differentiation of cells. Immunofluorescence was utilized to assess YAP expression levels for the purpose of determining the extent of mechanical stimulation in tissues.Results: Our findings showed that a few ossification lesions were detected behind the vertebral space of mice at 8 weeks of age. Spinal immobilization effectively restricts the flexion and extension of cervical and thoracic vertebrae in mice, delaying spinal ligament ossification and reducing chronic secondary spinal cord injury. Running exercises not only enhance the ossification area of the posterior longitudinal ligament (PLL) and Achilles tendons but also exacerbate secondary spinal cord injury. Further immunofluorescence results revealed a notable increase in YAP expression levels in tissues with severe ossification, suggesting that these tissues may be subjected to higher mechanical stimulation.Conclusion: Mechanical stimulation plays a pivotal role in the process of heterotopic ossification in tissues. Our study provided valid animal models to further explore the pathological mechanism of mechanical stimulation in HOTL development.

Abstract Probing endogenous molecules in living entities is significant to help decipher biological functions and exploit novel theranostics. DNA circuits that can recognize molecular inputs of interest and transduce them into readable signal outputs in an isothermal and autonomous manner have been actively pursued as versatile toolkits for intracellular biosensing research. Tremendous efforts are being devoted to developing integrated DNA circuits with high sensitivity, while spatiotemporal selectivity is often overlooked in the construction of functional DNA circuitry systems. This requires the development of stimuli‐responsive DNA circuits that can be activated on‐demand from the initial sensing‐blunt state to the sensing‐ready state under a programmable manner, for achieving precise bioimaging with high spatiotemporal control. In this review, an overview of recent advances in the construction of stimuli‐responsive DNA circuits that respond to particular triggers, including external physical stimuli and endogenous biological cues, and their spatiotemporally controllable molecular bioimaging applications is provided. The current challenges and potential solutions of these stimuli‐responsive DNA circuits for their future developments in this emerging field are also discussed.

Background: Intervertebral disc degeneration (IDD) is the leading cause of lower back pain, and an overall understanding of the molecular mechanisms related to IDD is still lacking. The purpose of this study was to explore gene signatures and immune cell infiltration related to IDD via bioinformatics analysis.Methods: A total of five expression profiles of mRNA and non-coding RNA were downloaded from the Gene Expression Omnibus (GEO) database. The potentially involved lncRNA/circRNA–miRNA–mRNA networks and protein-protein interaction networks were constructed by miRNet, circBank, STRING, and the Cytoscape database. Gene ontology, Kyoto Encyclopaedia of Genes and Genomes Analysis, Gene Set Enrichment Analysis, Gene Set Variation Analysis, Immune Infiltration Analysis, and Drug-Gene Interaction were used to analyse the top 20 hub genes. RT-qPCR was conducted to confirm the 12 differential expressions of genes both in the nucleus pulposus and annulus fibrosus tissuesResults: There were 346 differentially expressed mRNAs, 12 differentially expressed miRNAs, 883 differentially expressed lncRNAs, and 916 differentially expressed circRNAs in the GEO database. Functional and enrichment analyses revealed hub genes associated with platelet activation, immune responses, focal adhesion, and PI3K-Akt signalling. The apoptotic pathway, the reactive oxygen species pathway, and oxidative phosphorylation play an essential role in IDD. Immune infiltration analysis demonstrated that the Treg cells had significant infiltration, and three levels of immune cells, including dendritic cells, Th2 cells, and tumour-infiltrating lymphocytes, were inhibited in IDD. Drug-gene interaction analysis showed that COL1A1 and COL1A2 were targeted by collagenase clostridium histolyticum, ocriplasmin, and PDGFRA was targeted by 66 drugs or molecular compounds. Finally, 24 cases of IDD tissues and 12 cases of normal disc tissues were collected, and the results of RT-qPCR were consistent with the bioinformatics results.Conclusion: Our data indicated that the 20 hub genes and immune cell infiltration were involved in the pathological process of IDD. In addition, the PDGFRA and two potential drugs were found to be significant in IDD development.

Abstract Background Ossification of the posterior longitudinal ligaments (OPLL) is common disorder characterized by heterotopic ossification of the spinal ligaments. Mechanical stimulation (MS) plays an important role in OPLL. DLX5 is an essential transcription factor required for osteoblast differentiation. However, the role of DLX5 during in OPLL is unclear. This study aims to investigate whether DLX5 is associated with OPLL progression under MS. Methods Stretch stimulation was applied to spinal ligaments cells derived from OPLL (OPLL cells) and non‐OPLL (non‐OPLL cells) patients. Expression of DLX5 and osteogenesis‐related genes were determined by quantitative real‐time polymerase chain reaction and Western blot. The osteogenic differentiation ability of the cells was measured using alkaline phosphatase (ALP) staining and alizarin red staining. The protein expression of DLX5 in the tissues and the nuclear translocation of NOTCH intracellular domain (NICD) was examined by immunofluorescence. Results Compared with non‐OPLL cells, OPLL cells expressed higher levels of DLX5 in vitro and vivo (p

Blood always shows some immune changes after spinal cord injury (SCI), and detection of such changes in blood may be helpful for diagnosis and treatment of SCI. However, studies to date on blood immune changes after SCI in humans are not comprehensive. Therefore, to obtain the characteristics of blood immune changes and immunodiagnostic blood biomarkers of SCI and its different grades, a human blood transcriptome sequencing dataset was downloaded and analyzed to obtain differentially expressed immune-related genes (DEIGs), related functions and signaling pathways related to SCI and its various grades. Characteristic biomarkers of SCI and its different grades were identified by using weighted gene coexpression network analysis (WGCNA) and least absolute shrinkage and selection operator (LASSO) logistic regression. Expression of biomarkers was verified through experiments. The area under the curve (AUC) of biomarkers was calculated to evaluate their diagnostic value, and differences in immune cell content were examined. In this study, 17 kinds of immune cells with different contents between the SCI group and healthy control (HC) group were identified, with 7 immune cell types being significantly increased. Differences in the content of immune cells between different grades of SCI and the HC group were also discovered. DEIGs were identified, with alteration in some immune-related signaling pathways, vascular endothelial growth factor signaling pathways, and axon guidance signaling pathways. The SCI biomarkers identified and those of American Spinal Injury Society Impairment Scale (AIS) A and AIS D of SCI have certain diagnostic sensitivity. Analysis of the correlation of immune cells and biomarkers showed that biomarkers of SCI, AIS A grade and AIS D grade correlated positively or negatively with some immune cells. CKLF, EDNRB, FCER1G, SORT1, and TNFSF13B can be used as immune biomarkers for SCI. Additionally, GDF11and HSPA1L can be used as biomarkers of SCI AIS A grade; PRKCA and CMTM2 can be used as biomarkers of the SCI AIS D grade. Detecting expression of these putative biomarkers and changes in related immune cells may be helpful for predicting the severity of SCI.

Advancements in nanotechnology have promoted the rapid development of diverse nanosystems for biomedical applications. Manganese dioxide (MnO2) nanomaterials have attracted numerous attentions in the field of biosensing and cancer treatment due to their unique properties, such as simple fabrication procedures, broad optical absorption, and high oxidation and catalytic ability. Especially, the intelligent stimuli‐responsiveness of MnO2 nanomaterials to tumor microenvironment (TME) greatly improves their performance in tumor imaging and cancer therapy. In this review, the synthetic methods of different MnO2 nanostructures (nanoparticles, nanosheets, and nanoflowers) are first introduced, followed by the introduction of MnO2 nanomaterials for various biosensing applications based on their unique characteristics. Then, the recent achievements and progress of MnO2 nanomaterials for cancer treatment are comprehensively summarized with a focus on various therapeutic enhancements profiting from their disassembled ingredients under TME guidance. Finally, the current challenges and future perspectives of this promising research are discussed in the fields of biomedicine.

Poxvirus vectors represent versatile modalities for engineering novel vaccines and cancer immunotherapies. In addition to their oncolytic capacity and immunogenic influence, they can be readily engineered to express multiple large transgenes. However, the integration of multiple payloads into poxvirus genomes by traditional recombination-based approaches can be highly inefficient, time-consuming and cumbersome. Herein, we describe a simple, cost-effective approach to rapidly generate and purify a poxvirus vector with multiple transgenes. By utilizing a simple, modular CRISPR/Cas9 assisted-recombinant vaccinia virus engineering (CARVE) system, we demonstrate generation of a recombinant vaccinia virus expressing three distinct transgenes at three different loci in less than 1 week. We apply CARVE to rapidly generate a novel immunogenic vaccinia virus vector, which expresses a bacterial diadenylate cyclase. This novel vector, STINGPOX, produces cyclic di-AMP, a STING agonist, which drives IFN signaling critical to the anti-tumor immune response. We demonstrate that STINGPOX can drive IFN signaling in primary human cancer tissue explants. Using an immunocompetent murine colon cancer model, we demonstrate that intratumoral administration of STINGPOX in combination with checkpoint inhibitor, anti-PD1, promotes survival post-tumour challenge. These data demonstrate the utility of CRISPR/Cas9 in the rapid arming of poxvirus vectors with therapeutic payloads to create novel immunotherapies.

The therapeutic application of small interfering RNA (siRNA) is challenging due to its non-specific targeting and delivery issues. Here, the authors report an endogenous micro-RNA guided and hybridisation chain reaction-promoted siRNA delivery system encapsulated in tumour-derived extracellular vesicles, with cancer-specific activation, and achieve silencing of hypoxia-related genes.

Current DNA computational systems are constrained by integration efficiency, device structures and limited functions. Here the authors design a DNA arithmetic logic unit that uses polymerase-mediated strand displacement.

Berchemiella wilsonii (Rhamnaceae) is an endangered endemic tree to China. Here, the chloroplast genome was characterized by next-generation Illumina paired-end sequencing. The complete genome comprised 160,076 bp in length, with an inverted repeat region of 26,514 bp, a small single copy (SSC) region of 18,717 bp, and a large single copy region of 88,331 bp. A total of 132 genes were identified, including 83 protein-coding genes, 37 transfer RNA genes, and eight rRNA genes. The overall GC content of the whole genome was 37.23%. Phylogenetic analysis showed that B. wilsonii was most closely related to Berchemiella berchemiifolia.

The AC/DC hybrid microgrid has a large-scale and complex control process. It is of great significance and value to design a reasonable power coordination control strategy to maintain the power balance of the system. Based on hierarchical control, this paper designs a reasonable power coordination control strategy for AC/DC hybrid microgrid. For lower control, this paper designs a variety of control modes for each converter in different application scenarios. For the higher control, this paper analyzes the working mode of the system and designs the power coordination control strategy under the grid-connected and isolated island mode. In grid-connected operation, the DC bus voltage can be stabilized by adjusting the operation mode of the DC energy storage and the on-off of the secondary load. In isolated island operation, the DC sub-microgrid is the main microgrid, and the DC energy storage is the main power regulating equipment. This is based on the principle of “energy is in short supply in the system, DC energy storage finally discharge, energy supply exceeds demand in the system, DC energy storage gives priority to charging” of DC energy storage. By adjusting the control strategy of the micro-source, the reference power, and the on-off of the secondary load, the overall power balance is maintained. The Matlab/Simulink simulation software was used to build the AC/DC hybrid microgrid simulation model, which verified the effectiveness and stability of the proposed power coordination control strategy under various operating conditions.

The development of industry 4.0 has put forward higher requirements for modern milling technology. Monitoring the degree of milling tool wear during machine tool processing can improve product quality and reduce production losses. In the machining process of machine tools, many kinds of tools are usually used, and the signal characteristics of various sensors of different tools are different. Therefore, before the tool wear assessment, this paper identified the tool type according to the spindle current data. After the tool type recognition, this paper evaluates the tool wear degree according to the tool force data, vibration data, acoustic emission signal, and other multi-sensor data. Firstly, the Elman neural network and Adaboost algorithm are combined to construct the Elman_Adaboost strong predictor. Then, the variance and mean of seven sensor data were selected as the characteristic quantities to input the strong predictor. Finally, three wear quantities were obtained to measure the wear degree of the tool. The method proposed in this paper is implemented by Matlab, and the validity of this method is verified using the competition data provided by PHM (Prognostics and Health Management) Society. The results show that the average evaluation accuracy of the same tool wear is more than 92%, and that of the similar tool wear is more than 85%.

The sensing of pathogen infection and subsequent triggering of innate immunity are key to controlling zoonotic infections. Myxoma virus (MV) is a cytoplasmic DNA poxvirus that in nature infects only rabbits. Our previous studies have shown that MV infection of primary mouse cells is restricted by virus-induced type I interferon (IFN). However, little is known about the innate sensor(s) involved in activating signaling pathways leading to cellular defense responses in primary human immune cells. Here, we show that the complete restriction of MV infection in the primary human fibroblasts requires both tumor necrosis factor (TNF) and type I IFN. We also demonstrate that MV infection of primary human macrophages (pHMs) activates the cytoplasmic RNA sensor called retinoic acid inducible gene I (RIG-I), which coordinately induces the production of both TNF and type I IFN. Of note, RIG-I sensing of MV infection in pHMs initiates a sustained TNF induction through the sequential involvement of the downstream IFN-regulatory factors 3 and 7 (IRF3 and IRF7). Thus, RIG-I-mediated co-induction of TNF and type I IFN by virus-infected pHMs represents a novel innate defense mechanism to restrict viral infection in human cells. These results also reveal a new regulatory mechanism for TNF induction following viral infection.