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.

Treatment of lower cervical spine fracture with ankylosing spondylitis by simple long anterior cervical plate: a retrospective study of 17 cases.

Objective: Ankylosing spondylitis (AS), an autoimmune disease, often leads to lower cervical spine fractures, with the potential for severe spinal nerve damage even from low-energy injuries. The optimal treatment approach remains debated. Methods: A retrospective study involved 17 AS patients with lower cervical spine fractures who received anterior cervical fixation. Most presented cervicothoracic or thoracolumbar kyphosis, with 11 exhibiting neurological deficits. Patient characteristics, clinical data, visual analog scale (VAS), complications, and nerve recovery were analyzed. Results: No postoperative neurological deterioration occurred. All cases experienced complete fusion of fractures during the follow-up period. Preoperative VAS significantly decreased at 3 days and 3 months post-surgery. Of the 11 patients with preoperative neurological deficits, approximately 54.5% showed improvement post-surgery. No complications were reported, such as esophageal fistula, wound infection, or fixation failure. Conclusion: Anterior internal fixation is a possible treatment for AS-related lower cervical fractures. This approach ensures satisfactory spinal stability and neurological recovery with proper cranial traction and external fixation post-surgery. Our findings demonstrate that this surgical method is safe and effective.

Development and validation of a nomogram integrating marital status for 5-year overall survival of chondrosarcoma: a population-based study.

OBJECTIVES: The objective of this study was to evaluate the influence of marital status on overall survival (OS) and develop a nomogram for predicting 5-year OS in chondrosarcoma (CHS) patients. METHODS: We utilized the Surveillance, Epidemiology, and End Results (SEER) database to identify CHS patients diagnosed between 2010 and 2018. Survival rates were calculated using Kaplan-Meier analysis. Prognostic factors were identified through univariate and multivariate analyses. An independent cohort was used for external validation of the nomogram. Performance evaluation of the nomogram was conducted using Harrell's concordance index (C-index), calibration plot, and decision curve analysis (DCA). RESULTS: In the SEER cohort, Kaplan-Meier analysis showed significant differences in OS among CHS patients with different marital statuses (P < 0.001), with widowed patients having the lowest OS. In terms of gender, there were significant survival differences based on marital status in females (P < 0.001), but not in males (P = 0.067). The OS of married and single females is significantly higher than that of married (P < 0.001) and single male (P = 0.006), respectively. Kaplan-Meier curves showed no significant difference in OS between groups stratified by either gender or marital status in the external cohort. Univariate and multivariate analyses confirmed that age at diagnosis, gender, marital status, tumor size, histological type, tumor grade, SEER stage, and surgery were independent prognostic factors for OS. The nomogram demonstrated high internal and external validation C-indexes of 0.818 and 0.88, respectively. Calibration plots, DCA curve, and Kaplan-Meier curve (P < 0.001) confirmed the excellent performance and clinical utility of the nomogram. CONCLUSIONS: Marital status was an independent factor influencing OS in CHS patients, with widowed patients having the worst prognosis. The OS of both married and single females is significantly higher than that of their male counterparts. However, these findings require further validation in a large independent cohort. While the contribution of marital status on predicting OS appears modest, our nomogram accurately predicted 5-year OS and identified high-risk groups, providing a valuable tool for clinical decision-making.

The Role and Potential Mechanism of Complement Factor D in Fibromyalgia Development.

Purpose: Fibromyalgia (FM) is a syndrome characterized by chronic musculoskeletal pain. Its clinical symptoms include both somatic and psychiatric symptoms, making the treatment of FM extremely challenging. The cause of FM is still unknown, and some patients do not improve their symptoms even after long-term active treatment. Thus, the development of new targeted therapies is important to reduce pain and improve quality of life for FM patients. Methods: In this study, we screened genes and secreted factors that play key roles in FM through bioinformatics and big data analysis. Furthermore, we performed CCK-8, qRT-PCR, glucose, ATP and lactate content testing, dual luciferase reporter gene assay to investigate the potential mechanism of complement factor D in fibromyalgia development. Results: In bioinformatics and big data analysis, we identified CFD was negatively correlated with the pro-inflammatory factor IL-6 and positively correlated with the anti-inflammatory factor IL-4, which suggested that CFD may be an anti-inflammatory factor. Through cellular assays, we verified that CFD reversed the decrease in IL-4 expression and the increase in IL-6 expression in BV2 cells caused by ATP. Conclusion: In summary, based on bioinformatic methods and big data mining we obtained a new target CFD for FM, and further experiments verified that CFD has significant inhibition of ATP-induced neuropathic pain. These findings provide a new theoretical basis for the clinical translation of CFD.

Intraosseous regional prophylactic antibiotics decrease the risk of infection in total knee arthroplasty compared with intravenous antibiotics: a systematic review and meta-analysis.

Purpose: Surgical site infection (SSI) is a rare and serious complication of total knee arthroplasty (TKA), which causes a poor prognosis for patients. The purpose of this study was to explore the effect of intraosseous (IO) antibiotics in preventing infection and complications after TKA compared with intravenous (IV) antibiotics and to provide a certain theoretical basis for clinical treatment. Methods: The review process was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. We searched the PubMed, Embase, Ovid, Web of Science, and the Cochrane Central Register of Controlled Trials databases about trials on IO antibiotics (into the proximal tibia before skin incision) to prevent infections in TKA from the respective inception dates to September 30, 2022. The infection occurred within 3 months after surgery. Both researchers individually screened the studies in accordance with the inclusion and exclusion criteria, performed the literature quality evaluation and data extraction, and used Stata 17 software for data analysis. Results: Five studies that enrolled 3801 patients were included in this meta-analysis. The results showed that IO antibiotics were effective in reducing the incidence of SSI (OR: 0.25, P = 0.001) and periprosthetic joint infections (OR: 0.16, P = 0.004) relative to IV. Moreover, the percentage of infection due to Gram-positive bacteria (OR: 0.18, P = 0.025) was reduced in the IO group compared with that in IV group, but Gram-negative bacteria levels were not significantly reduced (P = 0.14). There was no difference between the two groups for other systemic adverse effects of the drug. Conclusions: IO antibiotics in TKA are safe and effective alternatives to IV antibiotics. Large randomized clinical studies comparing infection rates and related complications with IO and IV antibiotics are required.

Effect of the home care model in orthopedic wounds management.

Objective: To explore the effect of home care on orthopedic wound management. Materials and Methods: Patients with orthopedic wounds admitted from January 2020 to December 2022 were divided into a control group and a nursing group, with 23 cases in each group. After discharge, the control group was given routine health guidance and the nursing group was given home care. The mood score of the two groups of patients was evaluated, the time and cost were compared, and the patients' degree of trust and satisfaction with the medical staff were investigated. Results: The patients received home care had higher scores in mood, degree of trust and satisfaction with medical staff, and spent less time (P < .05). However, the cost was significantly higher than that of the control group (P < .05). Conclusions: Home care for patients with orthopedic wounds can significantly reduce the time spent on medical treatment. Moreover, improve patients' moods and trust and satisfaction with medical staff in a certain extent. However, better service often means more economic cost.

Delayed diagnosis of blunt thoracic aortic injury due to thoracic vertebral fracture: A case report and literature review.

Blunt vascular injury of the aorta combined with thoracolumbar fracture is rare. Delayed diagnosis may have a catastrophic outcome. We present a case of blunt thoracic aortic injury combined with a vertebral body fracture at T10 after a fall from height in which the diagnosis was delayed. After consultation with the vascular and spinal surgeons, we performed a thoracic endovascular aortic repair. When the patient's condition had stabilized, the fractures were reduced using posterior vertebral instrumentation. Prolonged compression of the thoracic aorta resulted in extensive necrosis of muscle tissues in the right lower leg. Fortunately, clinical and radiological examinations performed 7 months and 1 year later did not reveal any further damage. Cases of thoracic vertebral fracture with concomitant blunt thoracic aortic injury reported in the literature are reviewed. Thoracic endovascular aortic repair is a feasible, safe, and effective minimally invasive treatment for aortic injury when combined with thoracic vertebral fracture.

Acyloxyacyl hydrolase deficiency induces chronic inflammation and bone loss in male mice.

Hormonal homeostasis is essential in bone remodeling. Recent studies have shown that the treatment of intestinal inflammation can result in the regulation of bone resorption in distant bones. Increased intestinal permeability may lead to systemic inflammation and bone loss, also known as gut-bone axis. However, the underlying mechanism remains to be elucidated. Lipopolysaccharide (LPS) is a component of gram-negative bacteria that can increase osteoclastic differentiation in vitro. Acyloxyacyl hydrolase (AOAH) is a specific degrading enzyme of LPS, but little is known about the role of AOAH in bone metabolism. In this study, adult Aoah-/- mice showed a chronic inflammatory state and osteopenic phenotype analyzed by micro-CT and HE staining. Tartrate-resistant acid phosphatase (TRAP) staining of femurs showed an increase in TRAP-positive cells from Aoah-/- mice. AOAH depletion enhanced the osteoclast differentiation and bone resorption capacity of bone marrow-derived macrophages (BMMs). The enhanced osteoclast differentiation and bone resorption capacity of Aoah-/- BMMs were reversed by rAOAH. In conclusion, the chronic inflammatory state of adult Aoah-/- mice promotes bone resorption. AOAH participates in bone metabolism, which is mainly mediated by inhibiting osteoclast differentiation. LPS may be a key mediator of the gut-bone axis, and targeting AOAH may represent a feasible strategy for the treatment of chronic inflammatory bone resorption. KEY MESSAGES : AOAH knockout mice exhibited chronic inflammation mediated by LPS, and LPS may also serve as an important mediator in the regulation of bone metabolism in the gut-bone axis. AOAH regulated bone resorption by blocking the osteoclast differentiation via classical ERK and JNK pathways. rAOAH could rescue the enhanced osteoclast differentiation caused by AOAH deficiency.

Arthroscopy-Assisted Treatment of Wear Debris Pseudotumor After Total Knee Arthroplasty: A Case Report.

BACKGROUND: Wear debris pseudotumors are a rare complication after total knee arthroplasty (TKA) and have seldom been reported in the recent literature. There is no consensus on the best therapeutic method, but the high quality curative treatment, safe, low invasive treatments are required for the patients. CASE PRESENTATION: In this paper, we present the case of a 74-year-old man with a wear debris pseudotumor after TKA with symptoms of severe pain and functional disability of his right knee. X-ray examination showed that the medial compartment of the right knee was narrowing. Magnetic resonance imaging (MRI) and Doppler ultrasound both revealed a polycystic mass at the posteromedial side of the patient's right knee. Considering the bad health condition and the minimally invasive surgery requirement of this senior patient, arthroscopic knee debridement and percutaneous cystic mass suction were carried out simultaneously. Video arthroscopy of the right knee showed visible inflammatory soft tissue, obvious polyethylene fragments, wear of the polyethylene prothesis, and a broken polyethylene insert. The intraarticular polyethylene wear debris was removed as much as possible, and inflammatory soft tissue was debrided and sent for pathology. Postoperative pathology showed polyethylene debris in the soft tissue with an apparent multinucleated giant cell response, which was consistent with foreign body granuloma. All clinical manifestation was improved and Lysholm scores were significantly better at one year with this treatment, increasing from 32 points to 71 points. CONCLUSION: After two years of follow-up, the patient's knee joint was significantly relieved from soreness and pain, and walking was not significantly restricted. Our treatment could not address the root cause of the wear debris pseudotumor, which was due to prosthesis failure, but sometimes, such an approach is the safest, most economical, and most effective choice for patients who are intolerant to reoperation.

Genetic and pharmacological activation of Hedgehog signaling inhibits osteoclastogenesis and attenuates titanium particle-induced osteolysis partly through suppressing the JNK/c-Fos-NFATc1 cascade.

Rationale: Wear particle-induced periprosthetic osteolysis (PPO) is a common long-term complication of total joint arthroplasty, and represents the major cause of aseptic loosening and subsequent implant failure. Previous studies have identified the central role of osteoclast-mediated bone resorption in the pathogenesis of PPO. Thus, therapeutic approaches of inhibiting osteoclast formation and activity are considered to be of great potential to prevent and treat this osteolytic disease. Hedgehog (Hh) signaling has been shown to play an important role in promoting osteoblast differentiation and bone formation. While Hh signaling is also implicated in regulating osteoclastogenesis, whether it can directly inhibit osteoclast differentiation and bone resorption remains controversial. Moreover, its potential therapeutic effects on PPO have never been assessed. In this study, we explored the cell-autonomous role of Hh signaling in regulating osteoclastogenesis and its therapeutic potential in preventing wear particle-induced osteolysis. Methods: Hh signaling was activated in macrophages by genetically ablating Sufu in these cells using LysM-Cre or by treating them with purmorphamine (PM), a pharmacological activator of Smoothened (Smo). In vitro cell-autonomous effects of Hh pathway activation on RANKL-induced osteoclast differentiation and activity were evaluated by TRAP staining, phalloidin staining, qPCR analyses, and bone resorption assays. In vivo evaluation of its therapeutic efficacy against PPO was performed in a murine calvarial model of titanium particle-induced osteolysis by µCT and histological analyses. Mechanistic details were explored in RANKL-treated macrophages through Western blot analyses. Results: We found that Sufu deletion or PM treatment potently activated Hh signaling in macrophages, and strongly inhibited RANKL-induced TRAP+ osteoclast production, F-actin ring formation, osteoclast-specific gene expression, and osteoclast activity in vitro. Furthermore, we found that Sufu deletion or PM administration significantly attenuated titanium particle-induced osteoclast formation and bone loss in vivo. Our mechanistic study revealed that activation of Hh signaling suppressed RANKL-induced activation of JNK pathway and downregulated protein levels of two key osteoclastic transcriptional factors, c-Fos and its downstream target NFATc1. Conclusions: Both genetic and pharmacological activation of Hh signaling can cell-autonomously inhibit RANKL-induced osteoclast differentiation and activity in vitro and protect against titanium particle-induced osteolysis in vivo. Mechanistically, Hh signaling hinders osteoclastogenesis partly through suppressing the JNK/c-Fos-NFATc1 cascade. Thus, Hh signaling may serve as a promising therapeutic target for the prevention and treatment of PPO and other osteolytic diseases.

Corrigendum: Dihydroartemisinin prevents breast cancer-induced osteolysis via inhibiting both breast cancer cells and osteoclasts.

This corrects the article DOI: 10.1038/srep19074.

Dihydroartemisinin prevents breast cancer-induced osteolysis via inhibiting both breast caner cells and osteoclasts.

Bone is the most common site of distant relapse in breast cancer, leading to severe complications which dramatically affect the patients' quality of life. It is believed that the crosstalk between metastatic breast cancer cells and osteoclasts is critical for breast cancer-induced osteolysis. In this study, the effects of dihydroartemisinin (DHA) on osteoclast formation, bone resorption, osteoblast differentiation and mineralization were initially assessed in vitro, followed by further investigation in a titanium-particle-induced osteolysis model in vivo. Based on the proved inhibitory effect of DHA on osteolysis, DHA was further applied to MDA-MB-231 breast cancer-induced mouse osteolysis model, with the underlying molecular mechanisms further investigated. Here, we verified for the first time that DHA suppressed osteoclast differentiation, F-actin ring formation and bone resorption through suppressing AKT/SRC pathways, leading to the preventive effect of DHA on titanium-particle-induced osteolysis without affecting osteoblast function. More importantly, we demonstrated that DHA inhibited breast tumor-induced osteolysis through inhibiting the proliferation, migration and invasion of MDA-MB-231 cells via modulating AKT signaling pathway. In conclusion, DHA effectively inhibited osteoclastogenesis and prevented breast cancer-induced osteolysis.