Enzalutamide Sensitizes Castration-Resistant Prostate Cancer to Copper-Mediated Cell Death.

Despite the initial efficacy of enzalutamide in castration-resistant prostate cancer (CRPC), inevitable resistance remains a significant challenge. Here, the synergistic induction of copper-dependent cell death (cuproptosis) in CRPC cells is reported by enzalutamide and copper ionophores (elesclomol/disulfiram). Mechanistically, enzalutamide treatment increases mitochondrial dependence in CRPC cells, rendering them susceptible to cuproptosis, as evidenced by specific reversal with the copper chelator tetrathiomolybdate. This susceptibility is characterized by hallmarks of cuproptosis, including lipoylated protein aggregation and iron-sulfur cluster protein instability. Interestingly, the mitochondrial matrix reductase, FDX1, specifically correlates with elesclomol sensitivity, suggesting a potential mechanistic divergence between the two copper ionophores. Notably, this synergistic effect extends beyond in vitro models, demonstrating efficacy in 22Rv1 xenografts, mouse Pten p53 knockout organoids. Importantly, enzalutamide significantly enhances copper ionophore-mediated cytotoxicity in enzalutamide-resistant cells. Collectively, these findings indicate that enzalutamide and copper ionophores synergistically induce cuproptosis, offering a promising therapeutic avenue for CRPC, potentially including enzalutamide-resistant cases.

Sex dimorphism of IL-17-secreting peripheral blood mononuclear cells in ankylosing spondylitis based on bioinformatics analysis and machine learning.

BACKGROUND: Ankylosing spondylitis (AS) with radiographic damage is more prevalent in men than in women. IL-17, which is mainly secreted from peripheral blood mononuclear cells (PBMCs), plays an important role in the development of AS. Its expression is different between male and female. However, it is still unclear whether sex dimorphism of IL-17 contribute to sex differences in AS. METHODS: GSE221786, GSE73754, GSE25101, GSE181364 and GSE205812 datasets were collected from the Gene Expression Omnibus (GEO) database. Differential expressed genes (DEGs) were analyzed with the Gene Set Enrichment Analysis (GSEA), Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) methods. CIBERSORTx and EcoTyper algorithms were used for immune infiltration analyses. Machine learning based on the XGBoost algorithm model was used to identify the impact of DEGs. The Connectivity Map (CMAP) database was used as a drug discovery tool for exploring potential drugs based on the DEGs. RESULTS: According to immune infiltration analyses, T cells accounted for the largest proportion of IL-17-secreting PBMCs, and KEGG analyses suggested an enhanced activation of mast cells among male AS patients, whereas the expression of TNF was higher in female AS patients. Other signaling pathways, including those involving metastasis-associated 1 family member 3 (MAT3) or proteasome, were found to be more activated in male AS patients. Regarding metabolic patterns, oxidative phosphorylation pathways and lipid oxidation were significantly upregulated in male AS patients. In XGBoost algorithm model, DEGs including METRN and TMC4 played important roles in the disease process. we integrated the CMAP database for systematic analyses of polypharmacology and drug repurposing, which indicated that atorvastatin, famciclocir, ATN-161 and taselisib may be applicable to the treatment of AS. CONCLUSIONS: We analyzed the sex dimorphism of IL-17-secreting PBMCs in AS. The results showed that mast cell activation was stronger in males, while the expression of TNF was higher in females. In addition, through machine learning and the CMAP database, we found that genes such as METRN and TMC4 may promote the development of AS, and drugs such as atorvastatin potentially could be used for AS treatment.

Using GC-O-MS, GC-IMS, and chemometrics to investigate flavor component succession regularity in the Niulanshan Erguotou Baijiu brewing process.

The volatile compounds in Dacha liquor (DL) and Ercha liquor (EL) from Niulanshan Erguotou Baijiu (NEB) were analyzed. The results demonstrated that a total of 34 odorants were identified. For the first time, the products of different brewing stages were analyzed using temperature-programmed headspace gas chromatography-ion mobility spectrometry (TP-HS-GC-IMS). The 3D fingerprint obtained revealed that the compounds exhibited different change patterns during the brewing process. Furthermore, the results of principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) revealed that hexanal, 3-hydroxy-2-butanone, trans-2-pentenal, and ethyl hexanoate could be used to distinguish different types of fermented grains; and hexanal, 1-pentanol, methyl isovalerate, isoamyl acetate, 3-hydroxy-2-butanone, ethyl hexanoate, ethyl acetate, ethyl 2-methylbutanoate, and ethyl pentanoate could be used to distinguish different types of distilled spirits. This study serves as a useful reference for enhancing quality control measures in the production of NEB.

Intermittent administration of PTH for the treatment of inflammatory bone loss does not enhance entheseal pathological new bone formation.

OBJECTIVE: To investigate the effect of intermittent parathyroid hormone (iPTH) administration on pathological new bone formation during treatment of ankylosing spondylitis-related osteoporosis. METHODS: Animal models with pathological bone formation caused by hypothetical AS pathogenesis received treatment with iPTH. We determined the effects of iPTH on bone loss and the formation of pathological new bone with micro-computed tomography (micro-CT) and histological examination. In addition, the tamoxifen-inducible conditional knockout mice (CAGGCre-ERTM; PTHflox/flox, PTH-/-) was established to delete PTH and investigate the effect of endogenous PTH on pathological new bone formation. RESULTS: iPTH treatment significantly improved trabecular bone mass in the modified collagen-induced arthritis (m-CIA) model and unbalanced mechanical loading models. Meanwhile, iPTH treatment did not enhance pathological new bone formation in all types of animal models. Endogenous PTH deficiency had no effects on pathological new bone formation in unbalanced mechanical loading models. CONCLUSION: Experimental animal models of AS treated with iPTH show improvement in trabecular bone density, but not entheseal pathological bone formation，indicating it may be a potential treatment for inflammatory bone loss does in AS.

Complete resorption of the humerus in metastatic thyroid carcinoma: a case report.

BACKGROUND: Thyroid carcinoma is the most common endocrinological malignancy, but its spread to bone is rare. Particularly, bone metastases leading to complete resorption of the humerus are extremely uncommon. We aimed to explore factors affecting treatment decision in humeral metastasis by presenting a case and analyze the possible treatments via conducting a literature review. CASE PRESENTATION: We described a case of a 68-year-old woman experiencing chronic pain in her right upper arm for six years. Clinical, radiological, and pathological evaluations confirmed humeral metastasis from thyroid carcinoma. Surgical treatments like tumor removal or limb amputation were suggested for prolonging life and pain relief, but the patient refused them and pursued conservative managements such as herbal medicine, radioactive iodine (131I) therapy, and Levothyroxine Sodium(L-T4). The humeral destruction aggravated gradually, ultimately leading to complete resorption of her right humerus. The patient could not move her right shoulder, but her forearm motion was almost normal; thus, she could complete most of her daily living activities independently. Surgical treatments such as limb amputation were advised but she still refused them for preservation of the residual limb function and preferred conservative managements. CONCLUSION: A personalized multidisciplinary approach is important for patients with bone metastasis. The balance between limb amputation for life-prolonging and pain relief and limb salvage for preservation of residual function and social and psychological well-being should be considered. Our literature review revealed that some novel surgical treatments and techniques are available for bone metastases. This case adds to our current understanding of bone metastases and will contribute to future research and treatments.

Tenascin-C promotes endochondral ossification and fracture healing through Hedgehog and Hippo signaling.

Fractures are frequent and severe musculoskeletal injuries. This study aimed to investigate the function of tenascin-C (TNC) in regulating chondrogenic during fracture healing and elucidate the underlying molecular mechanisms. A well-established femur fracture model in male C57BL/6J mice was used to transect the middle diaphysis of the femur. To identify the essential role of TNC, shTNC lentiviruses or TNC protein were administered in the animal model. Micro-CT analysis, histologic analysis, immunostaining assays, and gene expression analysis were employed to investigate the effect of TNC during fracture healing. An in vitro mesenchymal stem cell culture system was developed to investigate the role and molecular mechanism of TNC in regulating chondrogenesis. TNC expression was induced at the inflammatory phase and peaked at the cartilaginous callus phase during fracture healing. Knockdown of TNC expression in callus results in decreased callus formation and impaired fracture healing. Conversely, administration of exogenous TNC promoted chondrogenic differentiation, cartilage template formation and ultimately improved fracture healing. Both the Hedgehog and Hippo signaling pathways were found to be involved in the pro-chondrogenic function of TNC. Our observations demonstrate that TNC is a crucial factor responsible for endochondral ossification in fracture healing and provide a potential therapeutic strategy for promoting fracture healing.

3D superimposition of human dentition contours in personal identification: A preliminary study.

The human permanent dentition has been commonly used for personal identification due to its uniqueness. Limited research, however, is conducted using 3D digital dental models. We propose to develop a new 3D superimposition method using the contours of human dentition and to further evaluate its feasibility. A total of 270 intraoral scan models were collected from 135 subjects. After a one-year interval, 52 subjects were chosen at random and the secondary intraoral scan models were obtained. The dentition contours of the first and secondary models were extracted to form a resource dataset and a test dataset. Through the application of the iterative nearest point (ICP) algorithm, the test dataset was registered with the resource dataset, and the root mean square error (RMSE) values of the point-to-point distances were calculated. 104 genuine pairs and 13,936 imposter pairs were generated, and in this study, the registration accuracy was 100%. The difference between mean RMSE values for the genuine pair (0.20 ± 0.06 mm) and the minimum RMSE value for the imposter pair (0.83 ± 0.06 mm) was significant in the maxillary arch (p < 0.05). Similarly, in the mandibular arch, the difference between mean RMSE values for the genuine pair (0.22 ± 0.07 mm) and the minimum RMSE value for the imposter pair (0.85 ± 0.08 mm) was significant (p < 0.05). The difference between the RMSE value for the genuine pair in the maxillary and the mandibular arch was significant (p < 0.05). This study indicated the feasibility of dentition contour-based model superimposition and could be considered for personal identification in the future.

Exploring the Role of Active Functional Microbiota in Flavor Generation by Integrated Metatranscriptomics and Metabolomics during Niulanshan Baijiu Fermentation.

Active functional microbiota for producing volatile flavors is critical to Chinese baijiu fermentation. Microbial communities correlated with the volatile metabolites are generally explored using DNA-based sequencing and metabolic analysis. However, the active functional microbiota related to the volatile flavor compounds is poorly understood. In this study, an integrated metatranscriptomic and metabolomics analysis was employed to unravel the metabolite profiles comprehensively and the contributing active functional microbiota for flavor generation during Niulanshan baijiu fermentation. A total of 395, 83, and 181 compounds were annotated using untargeted metabolomics, including LC-MS, GC-MS, and HS-SPME-GC-MS, respectively. Significant variances were displayed in the composition of compounds among different time-point samples according to the heatmaps and orthogonal partial least-square discriminant analysis. The correlation between the active microbiota and the volatile flavors was analyzed based on the bidirectional orthogonal partial least squares discriminant analysis (O2PLS-DA) model. Six bacterial genera, including Streptococcus, Lactobacillus, Pediococcus, Campylobacter, Yersinia, and Weissella, and five fungal genera of Talaromyces, Aspergillus, Mixia, Rhizophagus, and Gloeophyllum were identified as the active functional microbiota for producing the volatile flavors. In summary, this study revealed the active functional microbial basis of unique flavor formation and provided novel insights into the optimization of Niulanshan baijiu fermentation.

Unveiling innovation in aroma attribute evaluation of Niulanshan Baijiu: An advanced exploration of two different processing methods via food sensory omics and penalty analysis.

Niulanshan Baijiu (NLS), a notable variety of Baijiu known for its light flavor and extensive historical legacy, was subjected to a comparative analysis using two different processes: Hunzheng Xucha (HX) and Qingzheng Qingcha (QQ). The study combined sensory-oriented flavor analysis and penalty analysis to assess the differences between the two processes. Aroma compounds in NLS were extracted using liquid-liquid extraction and headspace solid phase microextraction. Gas chromatography-olfactometry-mass spectrometry was employed to identify 46 aroma-active compounds, including the first-time discovery of ethyl isohexanoate and 2,4-nonadienal in NLS. Quantification of 35 compounds with odor activity value (OAV) ≥ 1 was achieved using internal standard curve methods. Sensory assessments by a cohort of 111 participants highlighted the preference for HX-NLS in terms of flavor, while QQ-NLS exhibited a sour-Chen aroma that required improvement. The study further revealed the significant impact of acetic acid, butyric acid, hexanoic acid, octanoic acid, and 3-methylbutanal on the sour-Chen aroma in liquor.

Correlation Analysis and Diagnostic Value of Serum Homocysteine, Cystatin C and Uric Acid Levels with the Severity of Coronary Artery Stenosis in Patients with Coronary Heart Disease.

Background: Coronary angiography (CAG) is an invasive examination with high risks and costs and various complications may occur. It is necessary to find a diagnostic method, non-invasiveness, inexpensive with low risk. This study aims to analyze the correlation between the levels of serum homocysteine (Hcy), cystatin C (Cys C) and uric acid (UA) and Gensini score in patients with coronary heart disease (CHD) and assess their diagnostic value for CHD. Methods: A retrospective analysis was conducted on 1412 patients underwent CAG from October 2019 to December 2021, and we conducted this study from January to July 2022. A total of 765 patients with CHD confirmed by CAG were selected as the research group, while 647 patients revealed as non-obstructive stenosis by CAG as the control group. The serum Hcy, Cys C and UA levels were detected and the correlation between Gensini score and variables was analyzed. The receiver-operating characteristic (ROC) curve was performed to assess the diagnostic value of the Hcy, Cys C and UA for CHD. Results: The serum Hcy, Cys C and UA levels in the research group were higher as compared with the control group (p<0.05). Spearman correlation and multivariate linear regression analysis showed that there was a significantly positive correlation between Gensini score and serum Hcy, Cys C and UA levels (p<0.05). The ROC curve analysis presented the combined Hcy and Cys C with UA having the highest specificity of diagnostic value for CHD (area under the curve (AUC)=0.768, 95% CI 0.706-0.823, specificity = 72.34%, sensitivity = 67.88%, Youden Index = 0.4022). Conclusion: The serum Hcy, Cys C and UA levels in patients with CHD were significantly increased, positive correlation with Gensini score. The combined Hcy and Cys C with UA could be used to assess the severity of coronary artery stenosis and provide predictive and early intervention treatment values for CHD and a new way of diagnosing CHD, which is cheap, safe, effective and deserving of clinical application.

Cancer-keeper genes as therapeutic targets.

Finding cancer-driver genes has been a central theme of cancer research. We took a different perspective; instead of considering normal cells, we focused on cancerous cells and genes that maintained abnormal cell growth, which we named cancer-keeper genes (CKGs). Intervening CKGs may rectify aberrant cell growth, making them potential cancer therapeutic targets. We introduced control-hub genes and developed an efficient algorithm by extending network controllability theory. Control hub are essential for maintaining cancerous states and thus can be taken as CKGs. We applied our CKG-based approach to bladder cancer (BLCA). All genes on the cell-cycle and p53 pathways in BLCA were identified as CKGs, showing their importance in cancer. We discovered that sensitive CKGs - genes easily altered by structural perturbation - were particularly suitable therapeutic targets. Experiments on cell lines and a mouse model confirmed that six sensitive CKGs effectively suppressed cancer cell growth, demonstrating the immense therapeutic potential of CKGs.

Synthesis and Self-Assembly of Ultrathin Holey Graphdiyne Nanosheets for Oxygen Reduction Reaction.

Graphdiyne (GDY) is a fascinating graphene-like 2D carbon allotrope comprising sp and sp2 hybridized carbon atoms. However, GDY materials synthesized by solution-phase methods normally come as thick and porous films or amorphous powders with severely disordered stacking modes that obstruct macroscopic applications. Here, a facile and scalable synthesis of ultrathin holey graphdiyne (HGDY) nanosheets is reported via palladium/copper co-catalyzed homocoupling of 1,3,5-triethynylbenzene. The resulting freestanding 2D HGDY self-assembles into 3D foam-like networks which can in situ anchor clusters of palladium atoms on their surfaces. The Pd/HGDY hybrids exhibit high electrocatalytic activity and stability for the oxygen reduction reaction which outperforms that of Pt/C benchmark. Based on the ultrathin graphene-like sheets and their unique 3D interconnected macrostructures, Pd/HGDY holds great promise for practical electrochemical catalysis and energy-related applications.

The Effect and Possible Mechanism of Cardiac Rehabilitation in Partial Revascularization Performed on Multiple Coronary Artery Lesions.

Purpose: To observe the effect of cardiac rehabilitation (CR) in patients with partial revascularization performed on multiple coronary artery lesions and explore its possible mechanism. Patients and Methods: A total of 400 patients with multiple coronary artery lesions were enrolled and randomly divided into a complete revascularization group and a CR group, with 200 cases in each group. Target lesion revascularization was performed radically in the complete revascularization group, while it was partially completed in the CR group, and postoperative CR was performed. All the patients were put under conventional treatment. Left ventricular end diastolic dimension (LVEDD), left ventricular ejection fraction (LVEF), 6-minute walking distance (6-MWD), quality-of-life scores, safety and levels of serum nitric oxide (NO), nitric oxide synthase (NOS), superoxide dismutase (SOD), and vascular endothelial growth factor (VEGF) were evaluated and compared between two groups before and after training. Results: There was no significant difference in LVEDD, LVEF, 6-MWD, quality-of-life scores, levels of serum NO, NOS, SOD, and VEGF between two groups before training (p>0.05). 1 year later, compared with the complete revascularization group, the occurrence of major adverse events in the CR group declined (p>0.05); the measurements of LVEDD decreased and LVEF increased (p>0.05), 6-MWD increased significantly (p<0.05), quality-of-life scores were higher (p<0.05), the levels of serum NO, NOS, and SOD increased noticeably, and the levels of serum VEGF decreased significantly in the CR group (p<0.05). There were significant differences within the same group, before and after training (p<0.05). Conclusion: Cardiac rehabilitation training, not increase in the incidence of adverse events, is effective and safe after partial revascularization in patients with multiple coronary artery lesions, which has notable clinical advantages in promoting patients' exercise endurance and quality-of-life by improving the nitric oxide synthase system and antioxidant system and reducing the level of VEGF.

RNA-seq based elucidation of mechanism underlying the protective effect of Huangshui polysaccharide on intestinal barrier injury in Caco-2 cells.

Polysaccharides from Huangshui (HS) have the function of antioxidant and immunoregulation, but its intestinal barrier protection activity and the underlying mechanism remains unclear. The present work mainly studied the intestinal barrier protection function and its potential molecular mechanism of a heteropolysaccharide named NLS-2 with a molecular weight of 51.9 kDa. NLS-2 reduced intestinal permeability by decreasing the content of inflammatory cytokines and increasing the expression of tight junction (TJ) protein in LPS-damaged Caco-2 cells, thus protecting the intestinal barrier function. RNA-seq results showed that the differentially expressed genes (DEGs) were mainly enriched in the signaling pathways of MAPK, Toll-like receptor, and NF-κB. Subsequent western blot validation experiments proved that NLS-2 could indeed inhibit the two pathways of MAPK and NF-κB by reducing the expression of TRL4, thereby down-regulating the release of downstream pro-inflammatory cytokines and playing the role of intestinal barrier protection. Collectively, NLS-2 has beneficial effects on LPS-damaged intestinal barrier by inhibiting the TRL4/MyD88/NF-κB and MAPK signaling pathways.

Study on the mechanism of SALL4 down-regulation in promoting the invasion and migration of oral squamous cell carcinoma and influencing the survival and prognosis of patients.

Background: To investigate the identification of spalt-like transcription factor 4 (SALL4) in oral squamous cell carcinoma (OSCC). Methods: Recombinant cells loaded with miRNA expression cells were used to transform Tca8113 cells. Simple Tca8113 cells were used as the control group. We detected SALL4 messenger RNA (mRNA) before and after transfection by reverse transcription polymerase chain reaction (RT-PCR) and protein immunoblotting (western blot) A and protein expression. A dual luciferase reporter system was used to verify the targeted regulation of SALL4 and identify miRNA-S to test the effect of miRNA related to SALL4 regulation on the invasion and metastatic ability of Tca8113 cells. Results: The expression of SALL4 mRNA in Tca8113 cells was higher than that in the downregulated and control groups, respectively (P<0.05); there was no difference in Tca8113 cells between the upregulated and downregulated groups (P>0.05). Dual luciferase reporter system showed that the identified miRNA was miRNA-S; there were no differences in migration and invasion of Tca8113 cells between the up- and down-regulated groups (P>0.05). Conclusions: In human OSCC, SALL4 regulation-related miRNAs are poorly expressed and can inhibit the invasion and metastasis of tumor cells, which is expected to become a new therapeutic target for OSCC.

Metatranscriptomics Unravel Composition, Drivers, and Functions of the Active Microorganisms in Light-Flavor Liquor Fermentation.

The microbial community in the fermented pit determines the quantity and quality of light-flavor liquor. Genetic diversity and the potential functions of the microbial community are often analyzed by DNA-based omics sequencing. However, the features of the active microbial community have not been systematically studied. Here, metatranscriptomic analysis was performed to elucidate the active microbial composition, drivers, and their functions in light-flavor liquor fermentation. Bacterial genera, Lactobacillus, Streptococcus, Pediococcus, Thermotoga, and Faecalibacterium, and fungal genera, Saccharomyces, Talaromyces, Aspergillus, Clavispora, Rhizophagus, Cyberlindnera, and Wickerhamomyces, were the dominant active microorganisms during the fermentation process. Additionally, they dominated the three-stage fermentation successively. Redundancy analysis showed that pH, ethanol, moisture, and starch were the main driving forces of microbial succession. Among the genes for the respective carbohydrate-active enzyme families, those for the glycoside hydrolase family 23, the glycosyltransferase family 2, the carbohydrate-binding module family 50, the polysaccharide lyase family 4, the auxiliary activity family 1, and the carbohydrate esterase family 9 showed the highest expression level. Additionally, the highly expressed enzymes and their contributed microorganisms were found in the key KEGG pathways, including carbohydrate metabolism, energy metabolism, lipid metabolism, and amino acid metabolism. Based on these data, a functional model of carbohydrate hydrolysis, ethanol production, and flavor generation were proposed. Taken together, Saccharomyces, Lactobacillus, Wickerhamomyces, Pediococcus, Candida, and Faecalibacterium were suggested as the core active microorganisms. Overall, our findings provide new insights into the composition, drivers, and functions of the active microorganisms, which is crucial for improving the quality of light-flavor liquor. IMPORTANCE There is an urgent need for discovering the diversity and functions of the active microbial community in solid-state fermentation, especially in the pit of Chinese distilled liquor fermentation. Although the genetic composition of the microbial community has been clarified frequently by DNA-based sequencing, the composition and functions of the active microbial community have not been systematically revealed so far. Therefore, analysis of RNA-based data is crucial for discovering the functional microbial community. In this study, we employed metatranscriptomic analysis to elucidate the active microbial composition, successive drivers, and their functions in light-flavor liquor fermentation. The strategy can be broadly useful for discovering the active microbial community and exploring their functions in other types of flavor distilled liquor or other ecosystems. This study provides new insights into the understanding of the active microbial community composition and its functions.

CXCL12/CXCR4-Rac1-mediated migration of osteogenic precursor cells contributes to pathological new bone formation in ankylosing spondylitis.

Ankylosing spondylitis (AS) is a chronic inflammatory disease characterized by inflammatory back pain and spinal ankylosis due to pathological new bone formation. Here, we identified CXCL12 as a critical contributor to pathological new bone formation through recruitment of osteogenic precursor cells (OPCs). CXCL12 was found highly expressed in the regions that would potentially develop pathological new bone. OPCs were recruited to the regions where CXCL12 was up-regulated. Inhibition of CXCL12/CXCR4 axis with AMD3100 or conditional knockout of CXCR4 attenuated OPCs migration and subsequent pathological new bone formation in animal models of AS. By contrast, a genetically engineered animal model with CXCL12 overexpression developed a joint ankylosis phenotype. Furthermore, Rac1 was found essential for OPCs migration and pathological new bone formation. These findings ravel the novel role of CXCL12 in AS and indicate a potential strategy for targeting the CXCL12/CXCR4-Rac1 axis to prevent progression of axial skeleton ankylosis.

BRD4 promotes heterotopic ossification through upregulation of LncRNA MANCR.

AIMS: Acquired heterotopic ossification (HO) is a debilitating disease characterized by abnormal extraskeletal bone formation within soft-tissues after injury. The exact pathogenesis of HO remains unknown. It was reported that BRD4 may contribute to osteoblastic differentiation. The current study aims to determine the role of BRD4 in the pathogenesis of HO and whether it could be a potential target for HO therapy. METHODS: Achilles tendon puncture (ATP) mouse model was performed on ten-week-old male C57BL/6J mice. One week after ATP procedure, the mice were given different treatments (e.g. JQ1, shMancr). Achilles tendon samples were collected five weeks after treatment for RNA-seq and real-time quantitative polymerase chain reaction (RT-qPCR) analysis; the legs were removed for micro-CT imaging and subsequent histology. Human bone marrow mesenchymal stem cells (hBMSCs) were isolated and purified bone marrow collected during surgeries by using density gradient centrifugation. After a series of interventions such as knockdown or overexpressing BRD4, Alizarin red staining, RT-qPCR, and Western Blot (Runx2, alkaline phosphatase (ALP), Osx) were performed on hBMSCs. RESULTS: Overexpression of BRD4 enhanced while inhibition of Brd4 suppressed the osteogenic differentiation of hBMSCs in vitro. Overexpression of Brd4 increased the expression of mitotically associated long non-coding RNA (Mancr). Downregulation of Mancr suppressed the osteoinductive effect of BRD4. In vivo, inhibition of BRD4 by JQ1 significantly attenuated pathological bone formation in the ATP model (p = 0.001). CONCLUSION: BRD4 was found to be upregulated in HO and Brd4-Mancr-Runx2 signalling was involved in the modulation of new bone formation in HO. Cite this article: Bone Joint Res 2021;10(10):668-676.

Tenascin-C-mediated suppression of extracellular matrix adhesion force promotes entheseal new bone formation through activation of Hippo signalling in ankylosing spondylitis.

OBJECTIVES: The aim of this study was to identify the role of tenascin-C (TNC) in entheseal new bone formation and to explore the underlying molecular mechanism. METHODS: Ligament tissue samples were obtained from patients with ankylosing spondylitis (AS) during surgery. Collagen antibody-induced arthritis and DBA/1 models were established to observe entheseal new bone formation. TNC expression was determined by immunohistochemistry staining. Systemic inhibition or genetic ablation of TNC was performed in animal models. Mechanical properties of extracellular matrix (ECM) were measured by atomic force microscopy. Downstream pathway of TNC was analysed by RNA sequencing and confirmed with pharmacological modulation both in vitro and in vivo. Cellular source of TNC was analysed by single-cell RNA sequencing (scRNA-seq) and confirmed by immunofluorescence staining. RESULTS: TNC was aberrantly upregulated in ligament and entheseal tissues from patients with AS and animal models. TNC inhibition significantly suppressed entheseal new bone formation. Functional assays revealed that TNC promoted new bone formation by enhancing chondrogenic differentiation during endochondral ossification. Mechanistically, TNC suppressed the adhesion force of ECM, resulting in the activation of downstream Hippo/yes-associated protein signalling, which in turn increased the expression of chondrogenic genes. scRNA-seq and immunofluorescence staining further revealed that TNC was majorly secreted by fibroblast-specific protein-1 (FSP1)+fibroblasts in the entheseal inflammatory microenvironment. CONCLUSION: Inflammation-induced aberrant expression of TNC by FSP1+fibroblasts promotes entheseal new bone formation by suppressing ECM adhesion forces and activating Hippo signalling.

A pan-cancer analysis of molecular characteristics and oncogenic role of hexokinase family genes in human tumors.

Hexokinase (HK) plays a key role in various biological processes such as glycolysis of tumor cells. However, there is still a lack of systematic understanding of the contribution of HK family genes in different types of cancer. In the present study, we systematically analyzed the molecular changes and clinical correlations of HK family genes in 33 types of cancer extracted from more than 10,000 subjects. As a result, there were extensive genetic changes in HK family genes and the expression levels of HK family were significantly correlated with the activity of cancer marker-related pathways. In addition, HK family genes may be useful in predicting prognosis and therapeutic efficacy. Moreover, HK1,HK2 and HK3 may become potential oncogenes across a variety of cancer types. Furthermore, the oncogenic functions of HK1 in bladder cancer have been confirmed in vitro. Collectively, our results provide valuable resources to guide the mechanism and therapeutic analysis concerning the role of HK family genes in cancer.