Artificial intelligence reveals dysregulation of osteosarcoma and cuproptosis-related biomarkers, PDHA1, CDKN2A and neutrophils.

At present, the impact of cuproptosis-related genes in the study of osteosarcoma is largely unknown. Genome-wide data of osteosarcoma and controls were downloaded from 3 different databases, and specific diagnostic models associated with cuproptosis in osteosarcoma were constructed by support vector machines with artificial intelligence, random forest trees and LASSO regression. Differential analysis of immune cell infiltration was examined using routine blood data from 25,665 cases. Differential expression was examined using immunohistochemistry and PCR. PDHA1 and CDKN2A were obtained as specific cuproptosis-related biomarkers for osteosarcoma after artificial intelligence analysis. PDHA1, CDKN2A and neutrophils were differentially expressed in OS and control groups. PDHA1 and CDKN2A are significantly dysregulated in OS and are able to serve as biomarkers of OS.

Novel prognostic biomarkers, METTL14 and YTHDF2, associated with RNA methylation in Ewing's sarcoma.

Ewing's sarcoma has a poor prognosis and high metastasis rate; thus, it is critical to explore prognostic biomarkers of m6A-related genes. Two datasets were downloaded from the Gene Expression Omnibus database, m6A-related genes were extracted, and prognostic models were constructed using the least absolute shrinkage and selection operator and multivariate COX regression analyses. Immune cell composition and drug sensitivity analyses were performed, and our analysis was validated using laboratory methods of immunohistochemical specific staining and qRT-PCR. Ewing's sarcoma prognostic model demonstrated that the survival rate of cases in the high-risk group was much lower than that of the low-risk group. Naïve B cells, macrophages M0, macrophages M1, and resting mast cells are closely associated with Ewing's sarcoma. METTL14 and YTHDF2 are strongly associated with multiple drug sensitivity. Immunohistochemical specific staining revealed higher expression of both METTL14 and YTHDF2 in Ewing's sarcoma than in the paraneoplastic tissues. The results of qRT-PCR showed that METTL14 expression was significantly higher in both ES cell lines than in the control cell line. The prognostic model constructed using m6A-related genes METTL14 and TYHDF2, can be a potential prognostic biomarker for Ewing's sarcoma, with the survival rate of cases in the high-risk group being much lower than that of the low-risk group.

Comprehensive Network Analysis Identified SIRT7, NTRK2, and CHI3L1 as New Potential Markers for Intervertebral Disc Degeneration.

Intervertebral disc degeneration (IDD) is considered the basis of serious clinical symptoms, especially for low back pain (LBP). Therefore, it is essential to explore the regulatory role and diagnostic performance of dysregulated genes and potential drugs in IDD. Through WGCNA co-expression analysis, 36 co-expression modules were obtained. Among them, MidnightBlue and Red modules were the most related to IDD. Functional enrichment analysis showed that the Red module was mainly related to neutrophil activation and regulation of cytokine-mediated signaling pathway and apoptosis, whereas the MidnightBlue module was mainly related to extracellular matrix organization, bone development, extracellular matrix, extracellular matrix component, and other extracellular matrices. Furthermore, 356 genes highly related to the module were screened to construct a protein interaction network. Network degree distribution analysis showed that the known IDD-related genes had a higher degree of distribution. Enrichment analysis demonstrated that these genes were enriched in MAPK_SIGNALING_PATHWAY (FDR = 0.012), CHEMOKINE_SIGNALING_PATHWAY, and some other pathways. By constructing a disease-gene interaction network, three disease-specific genes were finally identified. Through combining with the drug-target gene interaction network, two potential therapeutic drugs, entrectinib and larotrectinib, were determined. Finally, based on these genes, the diagnostic model in the training dataset, test dataset, and verification dataset all showed a high diagnostic performance. The findings of this study contributed to the diagnosis of IDD and personalized treatment of IDD.

Role of AP-2α/TGF-ß1/Smad3 axis in rats with intervertebral disc degeneration.

OBJECTIVE: Studies have proposed the role of AP-2α in human disease. However, few have focused on its effects on intervertebral disc degeneration (IDD). This study intends to discuss the role of AP-2α in IDD by regulating TGF-ß1 and Smad3 expression. METHODS: The AP-2α and TGF-ß1 expression in IDD NP clinical samples was detected. Rat models of IDD were established by acupuncture. The rats were injected with AP-2α low expression adeno-associated virus or TGF-ß1 high expression adeno-associated virus to observe their effects on pathological damages, NP cell apoptosis, matrix metalloproteinase-2 (MMP-2), MMP-9, Smad3, Aggrecan and collagen (Col)-2 expression in NP tissues. The NP cells were isolated and transfected with silenced AP-2α or overexpressed TGF-ß1 vector to figure out their functions in growth, senescence and apoptosis. RESULTS: AP-2α and TGF-ß1 were upregulated in NP tissues of patients and rats with IDD. AP-2α silencing limited the activation of TGF-ß1 signaling pathway. Reduced AP-2α ameliorated pathological changes, declined MMP-2, MMP-9 and Smad3 expression and elevated Aggrecan and Col-2 expression in NP tissues of rats with IDD, and speeded up the growth and depressed senescence and apoptosis of NP cells of rats with IDD. Up-regulating TGF-ß1 weakened the effect of down-regulated AP-2α on NP tissues and cells in IDD. CONCLUSION: Collectively, our study demonstrates that knockdown of AP-2α restricts TGF-ß1 and Smad3 expression to promote proliferation and depress senescence and apoptosis of NP cells in rats with IDD.

Phenethyl isothiocyanate enhances adriamycin­induced apoptosis in osteosarcoma cells.

Adriamycin (ADM) is a first­line agent administered during the therapeutic regimes against osteosarcoma. Clinical administration of ADM produces systemic toxicity and resistance in patients, which restricts its applicability. In the present study the effects of phenethyl isothiocyanate (PEITC) on ADM­induced apoptosis in osteosarcoma cells was evaluated. Using U2­OS osteosarcoma cell line cells, treatment with PEITC or ADM for 24 h was observed to dose­dependently inhibit proliferation of U2­OS cells with half maximal inhibitory concentration (IC50) values of 5.33 µM and 10.32 µg/ml, respectively. When U2­OS cells were treated with a combination of the two agents, the inhibition was apparently enhanced, as the IC50 values decreased to 2 µM for PEITC and 1 µg/ml for ADM. Flow cytometry and terminal deoxynucleotidyl transferase dUTP nick end labeling revealed that treatment with PEITC or ADM alone reduced the viability of the U2­OS cells. Furthermore, the viability of the U2­OS cells was additionally reduced when treatment was with PEITC and ADM together. Supporting this finding, the activity and expression of caspase­3 were observed to be enhanced in the U2­OS cells following treatment with either PEITC or ADM, or a combination of the two. These results clearly indicate that PEITC enhances ADM­induced apoptosis in osteosarcoma cells.

Vanadate inhibits dexamethasone-induced apoptosis of rat bone marrow-derived mesenchymal stem cells.

Apoptosis of bone marrow-derived mesenchymal stem cells (BM-MSCs) has been shown to contribute to the development of osteoporosis, which is often the result of long-term use of glucocorticoid drugs such as dexamethasone (Dex). However, it remains unknown whether Dex induces apoptosis of BM-MSCs, and whether a chemical agent like vanadate can block such effects. To investigate these two issues, we isolated BM-MSCs from SD rats and treated the cells with different doses of Dex. We found that Dex induced apoptosis in dose- and time-dependent manners. Pretreating BM-MSCs with vanadate prevented Dex-induced apoptosis. Furthermore, we found that expression of caspases (3, 8, and 9) increased in Dex-treated BM-MSC and was attenuated by vanadate pretreatment. These results not only demonstrate the role of vanadate in the inhibition of Dex-induced apoptosis of BM-MSCs, but also reveal the therapeutic potential of vanadate in glucocorticoid-mediated osteoporosis.

Transcript levels of major interleukins in relation to the clinicopathological profile of patients with tuberculous intervertebral discs and healthy controls.

OBJECTIVES: The purpose of the present study was to simultaneously examine the transcript levels of a large number of interleukins (ILs; IL-9, IL-10, IL-12, IL-13, IL-16, IL-17, IL-18, IL-26, and IL-27) and investigate their correlation with the clinicopathological profiles of patients with tuberculous intervertebral discs. METHODS: Clinical data were collected from 150 patients participating in the study from January 2013 to December 2013. mRNA expression levels in 70 tuberculous, 70 herniated, and 10 control intervertebral disc specimens were determined by real-time polymerase chain reaction. RESULTS: IL-10, IL-16, IL-17, IL-18, and IL-27 displayed stronger expression in tuberculous spinal disc tissue than in normal intervertebral disc tissue (P<0.05). Our results illustrated multiple correlations among IL-10, IL-16, IL-17, IL-18, and IL-27 mRNA expression in tuberculous samples. Smoking habits were found to have a positive correlation with IL-17 transcript levels and a negative correlation with IL-10 transcript levels (P<0.05). Pain intensity, symptom duration, C-reactive protein levels, and the erythrocyte sedimentation rate exhibited multiple correlations with the transcript levels of several ILs (P<0.05). CONCLUSIONS: The experimental data imply a double-sided effect on the activity of ILs in tuberculous spinal intervertebral discs, suggesting that they may be involved in intervertebral discs destruction. Our findings also suggest that smoking may affect the intervertebral discs destruction process of spinal tuberculosis. However, further studies are necessary to elucidate the exact role of ILs in the intervertebral discs destruction process of spinal tuberculosis.

[Comparing hydroxyapatite coated versus non hydroxyapatite coated femoral stems in primary total hip arthroplasty: a meta analysis of randomized controlled trial].

OBJECTIVE: To evaluate the difference of clinical outcomes and radiological outcomes through meta-analysis on the total hip arthroplasty (THA) between hydroxyapatite(HA) coating and non-HA coating femoral stems. METHODS: We searched the MEDLINE, Embase, Cochrane library and CBM for published randomized controlled trial (RCT) comparing HA coating and non-HA coating femoral stems in primary THA clinical outcomes with Harris hip score and incidence postoperative thigh pain, radiological outcomes with presence of endosteal condensation and radioactive line on the prothesis, heterotopic ossification. Data analysis were performed using RevMan 5.0(the Cochrane Collaboration). RESULTS: Ten studies and 917 hips into our analysis, with 464 hips in HA groups and 453 hips in non-HA groups. The combined results of the meta-analysis indicated there was no statistical differences between the two groups on postoperative Harris hip score(WMD = 3.04, 95%CI:-4.47-10.54, P = 0.43) , there was statistical difference on incidence postoperative thigh pain (RR = 0.56, 95%CI:0.33-0.94, P = 0.03) . There were no significant differences between the two groups on presence of endosteal condensation (RR = 1.01, 95%CI:0.91-1.11, P = 0.91), presence of radioactive line (RR = 0.99, 95%CI:0.88-1.11, P = 0.83) and incidence of heterotopic ossification (RR = 0.97, 95%CI:0.77-1.21, P = 0.77). CONCLUSIONS: There are no clinical and radiological benefits in the use of HA coating femoral stems in Primary THA, there is not enough evidence prove the HA can reduce the incidence postoperative thigh pain.

Hydroxyapatite-coated femoral stems in primary total hip arthroplasty: a meta-analysis of randomized controlled trials.

BACKGROUND: Controversy exists over the use of hydroxyapatite (HA)-coated femoral stems in primary total hip arthroplasty (THA). We conducted a meta-analysis of randomized controlled trials (RCT) to compare the clinical and radiologic outcomes of primary THA using HA-coated versus non-HA-coated femoral stems. METHODS: Databases including MEDLINE, EMBASE and the Cochrane Library were searched to find relevant RCTs comparing HA-coated versus non-HA-coated femoral stems in primary THA. Data analyses were performed using RevMan 5.0 (The Cochrane Collaboration). RESULTS: Seven studies (792 hips) met the inclusion criteria. The pooled weighted mean difference (WMD) for the postoperative Harris hip score was 3.04 (95% CI: -4.47 to 10.54, P = 0.43). The cumulative risk ratios (RR) for the presence of endosteal condensation and radioactive lines were 1.02 (95% CI: 0.93 to 1.12, P = 0.64) and 1.01 (95% CI: 0.90 to 1.14, P = 0.81), respectively. CONCLUSIONS: This meta-analysis demonstrates that the use of HA-coated femoral stems in primary THA has no clinical or radiological benefits.

[Effect of associated ulnar styloid fracture on wrist function after distal radius].

OBJECTIVE: To evaluate the effect of associated ulnar styloid fracture on wrist function after distal radius fracture by comparing the clinical data between the cases of distal radius fracture with or without ulnar styloid fractures. METHODS: The clinical data of 182 patients with distal radius fracture between February 2005 and May 2010 were retrospectively analyzed, including 75 with ulnar styloid fracture (group A), and 107 without ulnar styloid fracture (group B). There was no significant difference in sex, age, disease duration, and fracture classification between 2 groups (P > 0.05). In groups A and B, closed reduction and splintlet or cast fixation were performed in 42 and 63 cases respectively, and open reduction and internal fixation in 33 and 44 cases respectively. All ulnar styloid fractures were not treated. RESULTS: The patients were followed up 21 months on average in group A and 20 months on average in group B. All incisions healed by first intention after operation. Ulnar wrist pain occurred in 4 patients (5.3%) of group A and 6 patients (5.6%) of group B, showing no significant difference (chi2=0.063, P=0.802). The fracture healing time was (10.9 +/- 2.7) weeks in group A and (11.6 +/- 2.3) weeks in group B, showing no significant difference (t=1.880, P=0.062). There was no significant difference in the palmar tilt angle, the ulnar inclination angle, and the radial length between groups A and B when fracture healing (P > 0.05). At last follow-up, there was no significant difference in wrist flexion-extension, radial-ulnar deviation, pronation-supination, and grip and pinch strength between 2 groups (P > 0.05). According to the Gartland-Werley score in groups A and B, the results were excellent in 24 and 35 cases, good in 43 and 57 cases, fair in 5 and 10 cases, and poor in 3 and 5 cases with execllent and good rate of 89.3% and 86.0%, respectively, showing no significant difference between 2 groups (Z=0.203, P=0.839). There were significant differences in the above indexes between patients undergoing closed reduction and open reduction in group A (P < 0.05). CONCLUSION: Associated ulnar styloid fracture has no obvious effect on the wrist function after distal radius fracture. The anatomical reduction of distal radial fracture is the crucial importance in the treatment of distal radial fracture accompanying ulnar styloid fracture.