Identifying autophagy-related mRNAs and potential ceRNA networks in meniscus degeneration based on RNA sequencing and experimental validation.

Purpose: The intimate connection between long noncoding RNA (lncRNA) and autophagy has been established in cartilage degeneration. However, their roles in meniscal degeneration remain ambiguous. This study aimed to identify the key autophagy-related lncRNA and its associated regulatory network in meniscal degeneration in the context of osteoarthritis (OA). Methods: RNA sequencing was performed to identify differentially expressed lncRNAs (DELs) and mRNAs (DEMs), which were then conducted to enrichment analyses using the DAVID database and Metascape. Autophagy-related DEMs were identified by combining DEMs with data from the Human Autophagy Database. Three databases were used to predict miRNA, and the DIANA LncBase Predicted database was utilized to predict miRNA-lncRNA interactions. Based on these predictions, comprehensive competitive endogenous RNA (ceRNA) network were constructed. The expression levels of the classical autophagy markers and autophagy-related ceRNA network were validated. Additionally, Gene Set Enrichment Analysis (GSEA) was performed using autophagy-related DEMs. Results: 310 DELs and 320 DEMs were identified, with five upregulated and one downregulated autophagy-related DEMs. Through reverse prediction of miRNA, paired miRNA-lncRNA interactions, and verification using RT-qPCR, two lncRNAs (PCAT19, CLIP1-AS1), two miRNA (has-miR-3680-3p and has-miR-4795-3p) and two mRNAs (BAG3 and HSP90AB1) were included in the constructed ceRNA regulatory networks. GSEA indicated that the increased expression of autophagy-related mRNAs inhibited glycosaminoglycan biosynthesis in the degenerative meniscus. Conclusion: This study presented the first construction of regulatory ceRNA network involving autophagy-related lncRNA-miRNA-mRNA interactions in OA meniscus. These findings offered valuable insights into the mechanisms underlying meniscal degeneration and provided potential targets for therapeutic intervention.

Seismic hazard prediction of the Hunhe Fault in the Shen-Fu New District.

Earthquake prevention and disaster mitigation are crucial aspects of social welfare that significantly impact national public security. This paper presents a seismic risk assessment and hazard prediction of the Hunhe Fault in the Shengyang-Fushun (Shen-Fu) New District. The target area is at risk of seismic damage due to two major branch ruptures, namely, F9 and F1; these ruptures have the potential to generate maximum earthquakes with a magnitude of 6.0 in the next 50 to 100 years. A three-dimensional underground velocity structure and asperity source model were established for the target faults. Subsequently, a hybrid technique combining deterministic and empirical approaches was employed to simulate the broadband strong ground motion of the target region in anticipation of the occurrence of expected scenario earthquakes. The distributions of peak ground acceleration (PGA), peak ground velocity (PGV) and peak ground displacement (PGD) for the area are provided, and the results indicate that densely populated urban areas could experience PGA values close to 280 cm/s2 along the fault traces. This study provides a reliable basis for engineering construction and urban planning in the Shen-Fu New District.

LTF induces senescence and degeneration in the meniscus via the NF-κB signaling pathway: A study based on integrated bioinformatics analysis and experimental validation.

Background: The functional integrity of the meniscus continually decreases with age, leading to meniscal degeneration and gradually developing into osteoarthritis (OA). In this study, we identified diagnostic markers and potential mechanisms of action in aging-related meniscal degeneration through bioinformatics and experimental verification. Methods: Based on the GSE98918 dataset, common differentially expressed genes (co-DEGs) were screened using differential expression analysis and the WGCNA algorithm, and enrichment analyses based on Gene Ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were further performed. Next, the co-DEGs were imported into the STRING database and Cytoscape to construct a protein‒protein interaction (PPI) network and further validated by three algorithms in cytoHubba, receiver operating characteristic (ROC) curve analysis and the external GSE45233 dataset. Moreover, the diagnostic marker lactotransferrin (LTF) was verified in rat models of senescence and replicative cellular senescence via RT‒qPCR, WB, immunohistochemistry and immunofluorescence, and then the potential molecular mechanism was explored by loss of function and overexpression of LTF. Results: According to the analysis of the GSE98918 dataset, we identified 52 co-DEGs (42 upregulated genes and 10 downregulated genes) in the OA meniscus. LTF, screened out by Cytoscape, ROC curve analysis in the GSE98918 dataset and another external GSE45233 dataset, might have good predictive power in meniscal degeneration. Our experimental results showed that LTF expression was statistically increased in the meniscal tissue of aged rats (24 months) and senescent passage 5th (P5) meniscal cells. In P5 meniscal cells, LTF knockdown inhibited the NF-κB signaling pathway and alleviated senescence. LTF overexpression in passage 0 (P0) meniscal cells increased the expression of senescence-associated secretory phenotype (SASP) and induced senescence by activating the NF-κB signaling pathway. However, the senescence phenomenon caused by LTF overexpression could be reversed by the NF-κB inhibitor pyrrolidine dithiocarbamate (PDTC). Conclusion: For the first time, we found that increased expression of LTF was observed in the aging meniscus and could induce meniscal senescence and degeneration by activating the NF-κB signaling pathway. These results revealed that LTF could be a potential diagnostic marker and therapeutic target for age-related meniscal degeneration.

Construction of miRNA-mRNA Regulatory Network to Identify Potential Biomarkers in Infantile Hemangioma by Integrated Bioinformatics Analysis.

Infantile hemangioma (IH) is the most common vascular tumor among infants and children. However, the understanding of pathogenesis about IH has not been fully elucidated, and the potential diagnostic maker remains further explored. In this study, we aimed to find miRNAs as potential biomarkers of IH through bioinformatic analysis. The microarray datasets GSE69136, GSE100682 were downloaded from the GEO database. The co-expressed differential miRNAs were identified by analyzing these two datasets. The downstream common target genes were predicted by the ENCORI, Mirgene, miRWalk, and Targetscan databases. GO annotation and KEGG pathway enrichment analysis for target genes were performed. The STRING database and Cytoscape software were used to construct the protein-protein interaction network and screen hub genes. Then potential diagnostic markers for IH were further screened and identified by using Receiver operating characteristic curve analysis. A total of thirteen co-expressed up-regulated miRNAs were screened out in the above two datasets, and 778 down-regulated target genes were then predicted. GO annotation and KEGG pathway enrichment analysis indicated that the common target genes strongly correlated with IH. Through the DEM-hub gene network construction, six miRNAs associated with the hub genes were identified. Finally, has-miR-522-3p, has-miR-512-3p, has-miR-520a-5p with high diagnostic values were screened out by receiver operating characteristic analysis. In the study, the potential miRNA-mRNA regulatory network was firstly constructed in IH. And, the three miRNAs might be used as potential biomarkers for IH, which also provided novel strategies for the therapeutic intervention of IH.

Immune Cell Infiltration Characteristics of Pigmented Villous Nodular Synovitis and Prediction of Potential Diagnostic Markers Based on Bioinformatics.

Background: Pigmented villous nodular synovitis (PVNS) is a tumor-like proliferative disease characterized by impairment of daily activities, decreased quality of life, and a high recurrence rate. However, the specific pathological mechanisms are still ill-defined and controversial. The purpose of this study was to define potential diagnostic markers and evaluate immune cell infiltration in the pathogenesis of PVNS. Method: The expression profile of GSE3698 was reanalyzed in the Gene Expression Omnibus (GEO) database. First, differentially expressed genes (DEGs) were identified using the R package "limma" and analyzed by Gene Ontology (GO) functional annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment. Next, the DEGs were imported into the STRING database and Cytoscape to construct a protein-protein interaction (PPI) network. Then, cytoHubba and ROC curve analyses were used to determine potential diagnostic biomarkers of PVNS. Finally, we used CIBERSORT to estimate the proportions of 22 immune cell subtypes in PVNS and analyzed the correlation between diagnostic markers and infiltrating immune cells. Result: We found 139 DEGs (including 93 upregulated genes and 46 downregulated genes). TYROBP, FCER1G, LAPTM5, and HLA-DPB1 were identified as potential diagnostic biomarkers of PVNS. Immune cell infiltration analysis indicated that neutrophils and M2 macrophages might be associated with the genesis and progression of PVNS. Furthermore, our correlation analysis of diagnostic markers and infiltrating immune cells found that TYROBP, FCER1G, LAPTM5, and HLA-DPB1 were positively correlated with M2 macrophage infiltration and that neutrophils, TYROBP, FCER1G, and LAPTM5 were negatively correlated with plasma cell infiltration. Conclusions: We identified TYROBP, FCER1G, LAPTM5, and HLA-DPB1 as potential diagnostic markers for PVNS and concluded that immune cell infiltration plays an important role in the genesis and progression of PVNS.

LINK-A lncRNA functions in the metastasis of osteosarcoma by upregulating HIF1α.

Long intergenic non-coding RNA for kinase activation (LINK-A) long non-coding RNA (lncRNA) is an oncogenic lncRNA in triple-negative breast cancer. The involvement of LINK-A lncRNA in other diseases is unknown. The present study aimed to investigate the possible involvement of LINK-A lncRNA in osteosarcoma. The results demonstrated that plasma levels of LINK-A lncRNA were significantly higher in patients with metastatic osteosarcoma (MO) compared with healthy controls and patients with non-metastatic osteosarcoma (NMO). LINK-A lncRNA overexpression significantly promoted cancer cell migration and invasion in osteosarcoma cell lines MG-63 and U20S. Upregulated expression of hypoxia-inducible factor 1α (HIF1α) was observed in cancer cells following LINK-A lncRNA overexpression. Exogenous HIF1α treatment did not significantly affect the expression of LINK-A lncRNA in cancer cells, whereas treatment with an HIF1α inhibitor significantly attenuated the effects of LINK-A lncRNA overexpression on cancer cell migration and invasion. Based on the results it was concluded that LINK-A lncRNA participated in the metastasis of osteosarcoma by upregulating HIF1α; upregulation of LINK-A lncRNA may serve as a potential diagnostic biomarker for patients with MO but not in those with NMO.

An Evidence-Based Meta-Analysis on the Roles of Functional Interleukin-6 Polymorphisms in Coronary Artery Disease.

Recently, the relationship between functional interleukin-6 (IL-6) polymorphisms and coronary artery disease (CAD) was extensively studied, with controversial findings. Therefore, we conducted this meta-analysis to better elucidate the relationship between these polymorphisms and the risk of CAD. A total of 57 case-control studies were finally included. The overall analyses showed that IL-6 -174G>C and -572G>C polymorphisms were significantly associated with the risk of CAD, the C allele of -174G>C (G versus C, odds ratio [OR] = 0.82, confidence interval [95% CI] = 0.75-0.89) and -572G>C polymorphisms (G versus C, OR = 0.82, 95% CI = 0.74-0.92) conferred an increased susceptibility to CAD. Further subgroup analyses yielded similar positive results for -174G>C polymorphism in Asian and Caucasian populations, and for -572G>C polymorphism in Asian and African populations. In conclusion, our findings suggest that IL-6 -174G>C and -572G>C polymorphisms may serve as potential genetic markers of CAD.

Surgical strategies for the treatment of os odontoideum with atlantoaxial dislocation.

OBJECTIVE This study aimed to compare the clinical results of using posterior fixation and fusion with or without anterior decompression to treat os odontoideum with atlantoaxial dislocation. METHODS Twenty-five consecutive patients with os odontoideum were included in this study. Sixteen patients with reducible atlantoaxial dislocation were treated by single-level posterior fusion and stabilization; the other 9 were treated with posterior fusion and stabilization combined with transoral decompression. Pre- and postoperative CT scans and MR images were obtained. RESULTS Twenty-four patients were followed for 24-54 months (average 36.5 months). Postoperative CT scans indicated that all pedicle screws were placed satisfactorily except in 2 cases, in which the screws slightly penetrated the transverse foramen. Postoperative MR images demonstrated that sufficient decompression of the spinal cord was obtained in all patients. Complications included 1 case each of pedicle screw breakage, pharynx ulcer, and persistent pharynx discomfort. Statistical analysis of all cases revealed that mean Japanese Orthopaedic Association scores improved from a preoperative score of 10.2 (range 7-13) to a postoperative score of 15.6 (range 11-18). CONCLUSIONS Patients who have os odontoideum with a reducible atlantoaxial dislocation can be effectively treated with single-level posterior fusion and stabilization. Combined transoral decompression and posterior fusion and stabilization is recommended for those with irreducible atlantoaxial dislocation.

Construction of conditional lentivirus-mediated shRNA vector targeting the human Mirk gene and identification of RNAi efficiency in rhabdomyosarcoma RD cells.

Rhabdomyosarcoma is the most common malignant soft tissue tumor in children. It has been demonstrated that Mirk as an activated protein kinase is overexpressed in rhabdomyosarcoma cells, which may be correlated with tumorigenesis. The aim of the present study was to explore the possibility of Mirk gene as a therapeutic target for the treatment of rhabdomyosarcoma, and the use of RNA interference in a temporally and spatially restricted manner to study the function of the target gene would be highly beneficial. To address this problem, a conditional lentivirus-mediated short hairpin RNA targeting human Mirk gene was constructed and employed to reduce endogenous Mirk expression in the rhabdomyosarcoma RD cell line in vitro. The expression of Mirk shRNA in RD cells transduced with this recombinant vector could be tracked with the expression of red fluorescent protein by the administration of doxycycline. A stable transgenic RD line was generated by transducing RD lines with the packaging viral particles. Quantitative PCR and western blot analysis indicated that the mRNA and protein levels of Mirk in the transgenic RD cells were significantly lower compared to those in the controls. In addition, the increasing apoptosis of RD cells induced by silencing of the Mirk gene was also observed. Overall, the results demonstrated that this recombinant vector-based RNAi expression system is an efficient approach to knockdown Mirk gene expression in the rhabdomyosarcoma RD cell line, which could, thereby, provide both a protocol to study the role of Mirk gene in tumor cells and a safer gene therapy in the clinic.