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1.
J Cell Biochem ; 119(6): 4548-4558, 2018 06.
Artigo em Inglês | MEDLINE | ID: mdl-29236326

RESUMO

Glia scar is a pathological marker in late phase of brain ischemia disease, which constitutes a major physical biochemical barrier to impede axonal regrowth. Astrocytes are known to be critically involved in the formation of glial scar. However, their response to ischemia and their role in neuroprotection after central nervous system (CNS) injury are not completely clear. Recently, we have demonstrated for the first time that Ski was up-regulated in reactive astrocytes after spinal cord injury in vivo and in vitro, which indicates Ski may be a new molecule that control astrocytes biologic properties after CNS injury. However, its role in the process of reactive astrogliosis after cerebral ischemia and its definite mechanism still remains unknown. This study is to elucidate the role of Ski in reactive astrocytes induced by oxygen-glucose deprivation/reoxygenation (OGD/R) model in vitro. The expression of Ski was proved to be up-regulated in OGD/R model. Meanwhile, Up-regulation of Ski was accompanied with high ratio of EdU (+) cells and up-expression of related proteins including GFAP, PCNA, CDK4, and CyclinD1, which demonstrated the distinct activation and proliferation of astrocytes after stimulation by OGD/R. Astrocytes were transfected with Ski-specific siRNA to knockdown Ski expression and subsequently attenuated OGD-induced astrocyte proliferation. Our results also showed that Ski down-regulation could suppress the activity of the Ras-Raf-ERK1/2 signaling pathway. Together, knockdown of Ski can effectively inhibit the proliferation of reactive astrogliosis via suppressing the Ras-Raf-ERK1/2 pathway. These findings indicated that maybe Ski is a promising therapeutic target for cerebral ischemic injury.


Assuntos
Astrócitos/metabolismo , Isquemia Encefálica/metabolismo , Proliferação de Células , Glucose/metabolismo , Sistema de Sinalização das MAP Quinases , Oxigênio/metabolismo , Proteínas Proto-Oncogênicas/deficiência , Animais , Astrócitos/patologia , Isquemia Encefálica/genética , Isquemia Encefálica/patologia , Hipóxia Celular , Técnicas de Silenciamento de Genes , Proteínas Proto-Oncogênicas/metabolismo , Ratos , Ratos Sprague-Dawley
2.
Biochem Biophys Res Commun ; 501(1): 55-63, 2018 06 18.
Artigo em Inglês | MEDLINE | ID: mdl-29678580

RESUMO

Fracture healing is a repair process of a mechanical discontinuity loss of force transmission, and pathological mobility of bone. Increasing evidence suggests that microRNA (miRNA) could regulate chondrocyte, osteoblast, and osteoclast differentiation and function, indicating miRNA as key regulators of bone formation, resorption, remodeling, and repair. Hence, during this study, we established a right femur fracture mouse model to explore the effect microRNA-185 (miR-185) has on osteoblasts in mice during fracture healing and its underlying mechanism. After successfully model establishment, osteoblasts were extracted and treated with a series of mimics or inhibitors of miR-185, or siRNA against PTH. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot analysis were performed to determine the levels of miR-185, PTH, ß-catenin and Wnt5b. Cell viability, cycle distribution and apoptosis were detected by means of MTT and flow cytometry assays. Dual luciferase reporter gene assay verified that PTH is a target gene of miR-185. Osteoblasts transfected with miR-185 mimics or siRNA against PTH presented with decreased levels of PTH, ß-catenin and Wnt5b which indicated that miR-185 blocks the Wnt/ß -catenin axis by inhibiting PTH. Moreover, miR-185 inhibitors promoted the osteoblast viability and reduced apoptosis with more cells arrested at the G1 stage. MiR-185 mimics were observed to have inhibitory effects on osteoblasts as opposed to those induced by miR-185 inhibitors. Above key results indicated that suppression of miR-185 targeting PTH could promote osteoblast growth and proliferation in mice during fracture healing through activating Wnt/ß -catenin axis.


Assuntos
Consolidação da Fratura/genética , MicroRNAs/genética , Osteoblastos/citologia , Osteoblastos/metabolismo , Hormônio Paratireóideo/antagonistas & inibidores , Hormônio Paratireóideo/genética , Animais , Apoptose/genética , Ciclo Celular/genética , Proliferação de Células/genética , Células Cultivadas , Regulação para Baixo , Consolidação da Fratura/fisiologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , MicroRNAs/agonistas , MicroRNAs/antagonistas & inibidores , Modelos Animais , Hormônio Paratireóideo/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Proteínas Wnt/genética , Proteínas Wnt/metabolismo , Via de Sinalização Wnt , beta Catenina/genética , beta Catenina/metabolismo
3.
J Arthroplasty ; 32(11): 3421-3428, 2017 11.
Artigo em Inglês | MEDLINE | ID: mdl-28662957

RESUMO

BACKGROUND: Controversy exists as to whether early functional outcomes differ after total hip arthroplasty performed using the direct anterior approach (DAA) or the posterolateral approach (PLA). METHODS: One hundred twenty patients were enrolled in this study and were divided into 2 groups based on surgical approach. Group A included patients who had a total hip arthroplasty with a DAA, whereas group B included those with a PLA. Patients were randomized into the DAA or PLA groups (n = 60), and perioperative and postoperative outcomes were recorded. RESULTS: When compared with the PLA, the DAA had a shorter incision length (9.1 vs 13.1 cm; P < .01), shorter hospital stay (2.8 vs 3.3 days, P = .04), and lower self-reported pain. Both serum inflammatory and muscle damage markers were lower in the DAA group. However, the PLA had shorter operative times (65.5 vs 83.3 min, P = .03) and less intraoperative blood loss (123.8 vs 165.9 mL, P = .04). The DAA had significantly lower variance in cup inclination and anteversion. Similar rates of intraoperative complications were identified in the 2 groups. The DAA was associated with better functional recovery at 3 months based on the Harris hip score, University of California Los Angeles activity score, and gait analysis; however, functional recovery at 6 months was similar between the 2 groups. CONCLUSION: We found functional advantages in early recovery after the DAA compared with the PLA. The DAA can offer rapid functional recovery with less muscle damage, greater pain relief, and lower variance in cup inclination and anteversion. However, no functional difference was found at 6 months follow-up.


Assuntos
Artroplastia de Quadril , Marcha , Recuperação de Função Fisiológica , Adulto , Idoso , Anestesia/métodos , Perda Sanguínea Cirúrgica/prevenção & controle , Celecoxib/uso terapêutico , China , Feminino , Humanos , Tempo de Internação , Masculino , Pessoa de Meia-Idade , Duração da Cirurgia , Período Pós-Operatório , Autorrelato , Índice de Gravidade de Doença , Resultado do Tratamento
5.
Mol Cell Biochem ; 406(1-2): 237-43, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25976667

RESUMO

Bone cells respond to various mechanical stimuli including fluid shear stress (FSS) in vitro. Induction of cyclooxygenase-2 (COX-2) is thought to be important for the anabolic effects of mechanical loading. Recently, extracellular-signal-regulated kinase 5 (ERK5) has been found to be involved in multiple cellular processes. However, the relationship between ERK5 and the induction of COX-2 is still unknown. Here, we investigated the potential involvement of ERK5 in the response of pre-osteoblastic MC3T3-E1 cells upon FSS. MC3T3-E1 cells were subjected to 12 dyn/cm(2) FSS. Then, we established a ERK5 small interfering RNA (siRNA) transfected cell line using the MC3T3-E1 cells. After the successful transfection confirmed by real-time reverse transcription-polymerase chain reaction and Western blotting, the expression of COX-2, cAMP response element-binding protein (CREB), and nuclear factor kappa B cells (NF-κB) were assayed for downstream effectors of activated ERK5 under FSS by Western blotting. Our results showed that FSS could stimulate COX-2 activity, and induce the phosphorylation of ERK5, CREB, and NF-κB. When the MC3T3-E1 cells were transfected using siRNA before exposure to FSS, COX-2 activity was suppressed, and the phosphorylation of CREB and NF-κB was significantly downregulated. In summary, we demonstrated that ERK5 pathway is essential in the induction of COX-2 gene.


Assuntos
Ciclo-Oxigenase 2/metabolismo , Proteína Quinase 7 Ativada por Mitógeno/metabolismo , Osteoblastos/enzimologia , Animais , Fenômenos Biomecânicos , Ciclo-Oxigenase 2/genética , Indução Enzimática , Camundongos , NF-kappa B/metabolismo , Células NIH 3T3 , Fosforilação , Processamento de Proteína Pós-Traducional , Transdução de Sinais , Estresse Fisiológico
6.
Clin Orthop Relat Res ; 473(8): 2672-9, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25981713

RESUMO

BACKGROUND: Arthroplasty has been shown to be superior regarding low risk of reoperation and better function score to internal fixation for treatment of displaced femoral neck fractures at short-term followup. However, there are unanswered questions regarding the efficacy of arthroplasty in the longer term compared with internal fixation. QUESTIONS/PURPOSES: We performed a meta-analysis comparing arthroplasty (hemiarthroplasty or THA) with internal fixation in patients with displaced femoral neck fractures with respect to (1) mortality, (2) reoperation, (3) functional recovery, and (4) complications, including only randomized trials with a minimum of 4 years followup. METHODS: Computerized databases, including PubMed (MEDLINE), EMBASE, Cochrane Register of Controlled Trials databases, and Web of Science™ were searched for studies published from the inception date for each database to March 2014. Eleven randomized controlled trials that compared arthroplasty (either hemiarthroplasty or THA) with internal fixation for treatment of patients with a femoral neck fracture were included in our analysis. The quality of the trials was assessed according to the Cochrane Handbook and meta-analyses were conducted using RevMan 5.2 software from the Cochrane Collaboration. The heterogeneity among studies was evaluated by the I-squared index (I2) and publication bias was assessed using forest plots. RESULTS: There were no differences between the internal fixation and arthroplasty groups for patient mortality at mid-term (48.4% vs 46.8%) or long-term followup (83.2% vs 81.5%). Arthroplasty was associated with a lower risk of reoperation at mid-term (7.2% vs 39.8%; relative risk [RR]=0.10; 95% CI, 0.06-0.07) and at long-term followup (14.3% vs 43.8%; RR=0.10; 95% CI, 0.06-0.07). Arthroplasty was associated with better functional recovery at mid-term followup (standard mean difference [SMD]=0.55; 95% CI, 0.02-1.09), whereas function at long-term followup (SMD=0.14; 95% CI, -0.35 to 0.62) was not different between the arthroplasty and internal fixation groups. There were no significant differences in subsequent ipsilateral fractures (1.5% vs 1.2%; RR=2.18; 95% CI, 0.32-14.67; p=0.42) and deep infections (2.7% vs 2.9%; RR=0.89; 95% CI, 0.40-2.01; p=0.78) between patients treated with arthroplasty and internal fixation. CONCLUSIONS: Based on our results, we found that compared with internal fixation, arthroplasty may result in a lower rate of subsequent reoperation at mid- and long-term followup, and better mid-term functional recovery. Future studies should investigate the mid- and long-term results of THAs compared with hemiarthroplasty.


Assuntos
Artroplastia de Quadril , Fraturas do Colo Femoral/cirurgia , Fixação Interna de Fraturas , Hemiartroplastia , Articulação do Quadril/cirurgia , Artroplastia de Quadril/efeitos adversos , Artroplastia de Quadril/mortalidade , Fraturas do Colo Femoral/diagnóstico , Fraturas do Colo Femoral/mortalidade , Fraturas do Colo Femoral/fisiopatologia , Fixação Interna de Fraturas/efeitos adversos , Fixação Interna de Fraturas/mortalidade , Hemiartroplastia/efeitos adversos , Hemiartroplastia/mortalidade , Articulação do Quadril/fisiopatologia , Humanos , Razão de Chances , Complicações Pós-Operatórias/mortalidade , Complicações Pós-Operatórias/cirurgia , Recuperação de Função Fisiológica , Reoperação , Fatores de Risco , Fatores de Tempo , Resultado do Tratamento
7.
Connect Tissue Res ; 55(2): 96-102, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24111522

RESUMO

The aim of this study was to determine the role of the mitogen-activated protein kinase kinase (MEK) 5/extracellular signal-regulated kinase (ERK) 5 pathway in osteoblast differentiation promoted by intermittent fluid shear stress (FSS). MC3T3-E1 osteoblastic cells were subjected to 12 dyn/cm(2) intermittent FSS, and the phenotypic markers for osteoblast differentiation, such as alkaline phosphatase (ALP) activity and expression of osteopontin (OPN) and osteocalcin (OCN), were then examined. The results showed that intermittent FSS could stimulate ERK5 phosphorylation, ALP activity and the expression of OPN and OCN. When the MEK5/ERK5 pathway was selectively inhibited by BIX02189, ALP activity was suppressed, and the expression of OPN and OCN was downregulated. Intermittent FSS induce the expression of Runt-related transcription factor-2 (Runx-2), which is involved in osteoblast differentiation by promoting the transcription of the above genes. Furthermore, the expression of Runx-2 was also reduced after treatment with BIX02189. Finally, we found that intermittent FSS was a more intense stimulus than steady FSS for promoting osteoblast differentiation. In summary, our results suggest that the MEK5/ERK5 pathway mediates osteoblast differentiation promoted by intermittent FSS, which was more effective than steady FSS in the differentiation process. The MEK5/ERK5 pathway also mediates FSS-induced Runx-2 expression in osteoblast differentiation.


Assuntos
Diferenciação Celular/fisiologia , MAP Quinase Quinase 5/metabolismo , Sistema de Sinalização das MAP Quinases/fisiologia , Proteína Quinase 7 Ativada por Mitógeno/metabolismo , Osteoblastos/enzimologia , Estresse Fisiológico/fisiologia , Animais , Antígenos de Diferenciação/biossíntese , Linhagem Celular , Regulação da Expressão Gênica/fisiologia , MAP Quinase Quinase 5/genética , Camundongos , Proteína Quinase 7 Ativada por Mitógeno/genética , Osteoblastos/citologia , Resistência ao Cisalhamento
8.
J Asian Nat Prod Res ; 16(6): 677-84, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24625202

RESUMO

A water-soluble polysaccharide (HPS3aS) with a molecular mass of 1.22 × 10(4) Da was isolated from Hedysarum polybotrys using anion-exchange and gel-permeation chromatography. HPS3aS exhibits a globular-shaped conformation in 0.1 M NaNO3 by size exclusion chromatography with multi-angle laser light scattering (SEC-MALLS). The investigation of the structural features of this heteropolysaccharide through a combination of chemical and instrumental analyses revealed that the backbone of HPS3aS is composed of α-D-(1 → 4)-linked glucopyranose residues, which occasionally branched at O-6. The branches are composed of (1 → 4)-linked galactopyranose residues and terminated with glucopyranose residues. HPS3aS possesses good in vitro antioxidant activity, as evaluated by scavenging assays with 1,1-diphenyl-2-picrylhydrazyl, hydroxyl, and superoxide radicals, which suggests that HPS3aS could be a potential antioxidant.


Assuntos
Antioxidantes/isolamento & purificação , Antioxidantes/farmacologia , Medicamentos de Ervas Chinesas/isolamento & purificação , Medicamentos de Ervas Chinesas/farmacologia , Fabaceae/química , Sequestradores de Radicais Livres/isolamento & purificação , Sequestradores de Radicais Livres/farmacologia , Polissacarídeos/isolamento & purificação , Polissacarídeos/farmacologia , Antioxidantes/química , Compostos de Bifenilo/farmacologia , Medicamentos de Ervas Chinesas/química , Sequestradores de Radicais Livres/química , Estrutura Molecular , Ressonância Magnética Nuclear Biomolecular , Oxirredução , Picratos/farmacologia , Raízes de Plantas/química , Polissacarídeos/química , Superóxidos/análise , Superóxidos/química , Água/química
9.
Orthop Surg ; 16(6): 1418-1433, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38658320

RESUMO

OBJECTIVE: Bone marrow mesenchymal stem cells (BMSCs) show significant potential for osteogenic differentiation. However, the underlying mechanisms of osteogenic capability in osteoporosis-derived BMSCs (OP-BMSCs) remain unclear. This study aims to explore the impact of YTHDF3 (YTH N6-methyladenosine RNA binding protein 3) on the osteogenic traits of OP-BMSCs and identify potential therapeutic targets to boost their bone formation ability. METHODS: We examined microarray datasets (GSE35956 and GSE35958) from the Gene Expression Omnibus (GEO) to identify potential m6A regulators in osteoporosis (OP). Employing differential, protein interaction, and machine learning analyses, we pinpointed critical hub genes linked to OP. We further probed the relationship between these genes and OP using single-cell analysis, immune infiltration assessment, and Mendelian randomization. Our in vivo and in vitro experiments validated the expression and functionality of the key hub gene. RESULTS: Differential analysis revealed seven key hub genes related to OP, with YTHDF3 as a central player, supported by protein interaction analysis and machine learning methodologies. Subsequent single-cell, immune infiltration, and Mendelian randomization studies consistently validated YTHDF3's significant link to osteoporosis. YTHDF3 levels are significantly reduced in femoral head tissue from postmenopausal osteoporosis (PMOP) patients and femoral bone tissue from PMOP mice. Additionally, silencing YTHDF3 in OP-BMSCs substantially impedes their proliferation and differentiation. CONCLUSION: YTHDF3 may be implicated in the pathogenesis of OP by regulating the proliferation and osteogenic differentiation of OP-BMSCs.


Assuntos
Biologia Computacional , Células-Tronco Mesenquimais , Osteogênese , Osteoporose Pós-Menopausa , Humanos , Osteoporose Pós-Menopausa/genética , Animais , Feminino , Células-Tronco Mesenquimais/metabolismo , Camundongos , Biologia Computacional/métodos , Osteogênese/fisiologia , Osteogênese/genética , Proteínas de Ligação a RNA/genética , Proteínas de Ligação a RNA/metabolismo , Aprendizado de Máquina , Diferenciação Celular , Adenosina/metabolismo , Adenosina/genética , Adenosina/análogos & derivados
10.
Cell Signal ; 115: 111038, 2024 03.
Artigo em Inglês | MEDLINE | ID: mdl-38195035

RESUMO

N6-methyladenosine (m6A), the most prevalent internal modification in mRNA, is related to the pathogenesis of osteoporosis (OP). Although methyltransferase Like-3 (METTL3), an m6A transferase, has been shown to mitigate OP progression, the mechanisms of METTL3-mediated m6A modification in osteoblast function remain unclear. Here, fluid shear stress (FSS) induced osteoblast proliferation and differentiation, resulting in elevated levels of METTL3 expression and m6A modification. Through Methylated RNA Immunoprecipitation Sequencing (MeRIP-seq) and Transcriptomic RNA Sequencing (RNA-seq), SRY (Sex Determining Region Y)-box 4 (SOX4) was screened as a target of METTL3, whose m6A-modified coding sequence (CDS) regions exhibited binding affinity towards METTL3. Further functional experiments demonstrated that knockdown of METTL3 and SOX4 hampered osteogenesis, and METTL3 knockdown compromised SOX4 mRNA stability. Via RNA immunoprecipitation (RIP) assays, we further confirmed the direct interaction between METTL3 and SOX4. YTH N6-Methyladenosine RNA Binding Protein 3 (YTHDF3) was identified as the m6A reader responsible for modulating SOX4 mRNA and protein levels by affecting its degradation. Furthermore, in vivo experiments demonstrated that bone loss in an ovariectomized (OVX) mouse model was reversed through the overexpression of SOX4 mediated by adeno-associated virus serotype 2 (AAV2). In conclusion, our research demonstrates that METTL3-mediated m6A modification of SOX4 plays a crucial role in regulating osteoblast proliferation and differentiation through its recognition by YTHDF3. Our research confirms METTL3-m6A-SOX4-YTHDF3 as an essential axis and potential mechanism in OP.


Assuntos
Metiltransferases , Osteoblastos , Animais , Camundongos , Proliferação de Células , Metiltransferases/metabolismo , Osteoblastos/metabolismo , RNA , RNA Mensageiro/metabolismo
11.
Zhongguo Gu Shang ; 36(10): 926-31, 2023 Oct 25.
Artigo em Chinês | MEDLINE | ID: mdl-37881923

RESUMO

OBJECTIVE: To compare the posterior cruciate ligament(PCL) index with six different measurement methods, and analyze and verify its clinical diagnostic value in anterior cruciate ligament (ACL) injury. METHODS: The Magnetic resonance imaging (MRI) data of 225 knee joints in our hospital from May 2018 to March 2022 were retrospectively analyzed, aged from 18 to 60 years old, with a median of 32 years old. On the sagittal MRI images of 114 patients with ACL injury and 111 patients with intact ACL, Measure the straight-line distance (A) between the femoral attachment point and the tibial attachment point of the PCL on the MRI sagittal image and the maximum vertical distance (B) between the straight line and the arcuate mark point of the PCL on the sagittal image, calculate the PCL index and evaluate the diagnostic value of the PCL index for ACL injury. RESULTS: The PCL index of the ACL normal group and the ACL injury group were statistically described. There was no significant difference in PCL index 1, 2, 3 and 6 between the two groups(P>0.05). The difference of PCL index 4 and 5 between the two groups was statistically significant (P<0.001). This study only found that the PCL index 2, 6 in the ACL normal group had a negative correlation with the patient's age (correlation coefficient=-0.213, -0.819;P<0.05), and the PCL index 5 in the ACL injury group was significantly correlated with the patient's body mass index(BMI)had a negative correlation (correlation coefficient=-0.277, P<0.05). CONCLUSION: The change of PCL index is helpful for the diagnosis of ACL injury, PCL index 4 and 5 can be used as effective reference indexes for diagnosing ACL injury in clinic.


Assuntos
Lesões do Ligamento Cruzado Anterior , Ligamento Cruzado Posterior , Humanos , Adolescente , Adulto Jovem , Adulto , Pessoa de Meia-Idade , Ligamento Cruzado Posterior/diagnóstico por imagem , Lesões do Ligamento Cruzado Anterior/diagnóstico por imagem , Ligamento Cruzado Anterior , Estudos Retrospectivos , Articulação do Joelho , Imageamento por Ressonância Magnética/métodos
12.
Heliyon ; 9(1): e12799, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36699262

RESUMO

Background: Rheumatoid arthritis (RA) is an autoimmune disease that affects individuals of all ages. The basic pathological manifestations are synovial inflammation, pannus formation, and erosion of articular cartilage, bone destruction will eventually lead to joint deformities and loss of function. However, the specific molecular mechanisms of synovitis tissue in RA are still unclear. Therefore, this study aimed to screen and explore the potential hub genes and immune cell infiltration in RA. Methods: Three microarray datasets (GSE12021, GSE55457, and GSE55235), from the Gene Expression Omnibus (GEO) database, have been analyzed to explore the potential hub genes and immune cell infiltration in RA. First, the LIMMA package was used to screen the differentially expression genes (DEGs) after removing the batch effect. Then the clusterProfiler package was used to perform functional enrichment analyses. Second, through weighted coexpression network analysis (WGCNA), the key module was identified in the coexpression network of the gene set. Third, the protein-protein interaction (PPI) network was constructed through STRING website and the module analysis was performed using Cytoscape software. Fourth, the CIBERSORT and ssGSEA algorithm were used to analyze the immune status of RA and healthy synovial tissue, and the associations between immune cell infiltration and RA-related diagnostic biomarkers were evaluated. Fifth, we used the quantitative reverse transcription-polymerase chain reaction (qRT-PCR) to validate the expression levels of the hub genes, and ROC curve analysis of hub genes for discriminating between RA and healthy tissue. Finally, the gene-drug interaction network was constructed using DrugCentral database, and identification of drug molecules based on hub genes using the Drug Signature Database (DSigDB) by Enrichr. Results: A total of 679 DEGs were identified, containing 270 downregulated genes and 409 upregulated genes. DEGs were primarily enriched in immune response and chemokine signaling pathways, according to functional enrichment analysis of DEGs. WGCNA explored the co-expression network of the gene set and identified key modules, the blue module was selected as the key module associated with RA. Seven hub genes are identified when PPI network and WGCNA core modules are intersected. Immune infiltration analysis using CIBERSORT and ssGSEA algorithms revealed that multiple types of immune infiltration were found to be upregulated in RA tissue compared to normal tissue. Furthermore, the levels of 7 hub genes were closely related to the relative proportions of multiple immune cells in RA. The results of the qRT-PCR demonstrated that the relative expression levels of 6 hub genes (CD27, LCK, CD2, GZMB, IL7R, and IL2RG) were up-regulated in RA synovial tissue, compared with normal tissue. Simultaneously, ROC curves indicated that the above 6 hub genes had strong biomarker potential for RA (AUC >0.8). Conclusions: Through bioinformatics analysis and qRT-PCR experiment, our study ultimately discovered 6 hub genes (CD27, LCK, CD2, GZMB, IL7R, and IL2RG) that closely related to RA. These findings may provide valuable direction for future RA clinical diagnosis, treatment, and associated research.

13.
Mol Cell Biochem ; 364(1-2): 321-7, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-22286747

RESUMO

Fluid shear stress plays an important role in bone remodeling, however, the mechanism of mechanotransduction in bone tissue remains unclear. Recently, ERK5 has been found to be involved in multiple cellular processes. This study was designed to investigate the potential involvement of ERK5 in the proliferative response of osteoblastic cells to cyclic fluid shear stress. We reported here that cyclic fluid shear stress promoted ERK5 phosphorylation in MC3T3-E1 cells. Inhibition of ERK5 phosphorylation attenuated the increased expression of AP-1 and cyclin D1 and cell proliferation induced by cyclic fluid flow, but promoted p-16 expression. Further more, we found that cyclic fluid shear stress was a better stimuli for ERK5 activation and cyclin D1 expression compared with continuous fluid shear stress. Moreover, the pharmacological ERK5 inhibitor, BIX02189, which inhibited ERK5 phosphorylation in a time-dependent manner and the suppression lasted for at least 4 h. Taken together, we demonstrate that ERK5/AP-1/cyclin D1 pathway is involved in the mechanism of osteoblasts proliferation induced by cyclic fluid shear stress, which is superior in promoting cellular proliferation compared with continuous fluid shear stress.


Assuntos
Proliferação de Células , Ciclina D1/metabolismo , Regulação da Expressão Gênica/genética , Proteína Quinase 7 Ativada por Mitógeno/metabolismo , Osteoblastos/metabolismo , Compostos de Anilina/farmacologia , Animais , Linhagem Celular , Ciclina D1/genética , Quinase 4 Dependente de Ciclina/metabolismo , Inibidor p16 de Quinase Dependente de Ciclina/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Peróxido de Hidrogênio/farmacologia , Indóis/farmacologia , Camundongos , Proteína Quinase 7 Ativada por Mitógeno/genética , Osteoblastos/enzimologia , Fosforilação/efeitos dos fármacos , Transdução de Sinais , Estresse Mecânico , Fator de Transcrição AP-1/genética , Fator de Transcrição AP-1/metabolismo
14.
Cell Biol Int ; 36(3): 229-36, 2012 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-21954859

RESUMO

Mechanotransduction is a complicated process, of which mechanosensation is the first step. Previous studies have shown that the cytoskeleton plays a crucial role in mechanosensation and the mediation of intracellular signal transduction. However, the mechanism of mechanotransduction in the bone remains elusive. Here, we investigated the potential involvement of a novel MAPK (mitogen-activated protein kinase) member, ERK5 (extracellular-signal-regulated kinase 5), in the response of osteoblastic cells to FSS (fluid shear stress). Our results demonstrated that ERK5 was rapidly phosphorylated in pre-osteoblastic MC3T3-E1 cells upon FSS, and the integrity and reorganization of the cytoskeleton were critical in this process, in which the cytoskeleton-dependent activation of FAK (focal adhesion kinase) may be involved in the activation of ERK5 induced by FSS. Moreover, we found that cytoskeletal disruption led to significant down-regulation of ERK5 phosphorylation, but had no effect on ERK5 nuclear localization. Furthermore, the cytoskeleton rapidly reorganized in response to FSS, but long-time fluid load, even at a physiological level, led to cytoskeletal disruption, suggesting that other pathways may be involved in long-term mechanotransduction. Taken together, our data provide new insight into the mechanisms of mechanosensation by highlighting the link between ERK5 activation and cytoskeletal reorganization in osteoblasts undergoing FSS.


Assuntos
Citoesqueleto/metabolismo , Proteína Quinase 7 Ativada por Mitógeno/metabolismo , Osteoblastos/metabolismo , Estresse Mecânico , Animais , Células Cultivadas , Regulação para Baixo , Mecanotransdução Celular , Camundongos , Proteína Quinase 7 Ativada por Mitógeno/genética
15.
Artigo em Inglês | MEDLINE | ID: mdl-20653335

RESUMO

OBJECTIVE: To investigate the biocompatibility of diamond-like carbon (DLC) coated nickel-titanium shape memory alloy (NiTi SMA) in vitro and in vivo. METHODS: The in vitro study was carried out by co-culturing the DLC coated and uncoated NiTi SMA with bone marrow mesenchymal stem cells (MSCs), respectively, and the in vivo study was carried out by fixing the rabbits' femoral fracture model by DLC coated and uncoated NiTi SMA embracing fixator for 4 weeks, respectively. The concentration of the cells, alkaline phosphatase (AKP), and nickel ion in culture media were detected, respectively, at the first to fifth day after co-culturing. The inorganic substance, osteocalcin, alkaline phosphatase (ALP), and tumor necrosis factor (TNF) in callus surrounding fracture and the Ni(+) in muscles surrounding fracture site, liver and brain were detected 4 weeks postoperatively. RESULTS: The in vitro study showed that the proliferation of MSCs and the expression of AKP in the DLC-coated group were higher than the uncoated group (P < 0.05), while the uncoated group released more Ni(2+) into the culture media than that in the coated group (P < 0.05). The in vivo study revealed that the inorganic substance and AKP, osteocalcin, and TNF expression were significantly higher in the DLC coated NiTi SMA embracing fixator than that in the uncoated group (P < 0.05). Ni(2+) in liver, brain, and muscles surrounding the fracture were significantly lower in the DLC coated groups than that in the uncoated group (P < 0.05). CONCLUSION: Nickel-titanium shape memory alloy coated by diamond-like carbon appears to have better biocompatibility in vitro and in vivo compared to the uncoated one.


Assuntos
Compostos Inorgânicos de Carbono/administração & dosagem , Materiais Revestidos Biocompatíveis/administração & dosagem , Fraturas do Fêmur/terapia , Células-Tronco Mesenquimais/efeitos dos fármacos , Níquel/administração & dosagem , Titânio/administração & dosagem , Fosfatase Alcalina/metabolismo , Animais , Medula Óssea/patologia , Compostos Inorgânicos de Carbono/efeitos adversos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Materiais Revestidos Biocompatíveis/efeitos adversos , Fraturas do Fêmur/metabolismo , Fraturas do Fêmur/cirurgia , Humanos , Teste de Materiais , Células-Tronco Mesenquimais/metabolismo , Células-Tronco Mesenquimais/patologia , Músculos/efeitos dos fármacos , Músculos/metabolismo , Músculos/patologia , Níquel/efeitos adversos , Níquel/metabolismo , Dispositivos de Fixação Ortopédica/efeitos adversos , Osteocalcina/metabolismo , Coelhos , Titânio/efeitos adversos , Fator de Necrose Tumoral alfa/metabolismo
16.
Orthop Surg ; 13(1): 328-337, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33426744

RESUMO

OBJECTIVE: To compare the clinical efficacy of percutaneous full-endoscopic transforaminal lumbar interbody fusion (Endo-TLIF) with percutaneous pedicle screws (PPSs) performed by using a visualization system with that of minimally invasive transforaminal lumbar interbody fusion (MIS-TLIF) for the treatment of degenerative lumbar spinal stenosis (LSS). METHODS: From June 2017 to May 2018, the data of a total of 78 patients who met the selection criteria were retrospectively reviewed and were divided into the Endo-TLIF group (40 cases) and the MIS-TLIF group (38 cases) according to the surgical method used. The visual analog scale (VAS) and the Japanese Orthopaedic Association (JOA) scale were administered preoperatively and at the 1-week, 3-month, and 1-2-year follow-ups. The fusion rate and major complications, including revision, were also recorded. RESULTS: All the patients were followed up for 24 to 34 months, with an average follow-up of 30.7 months. The intraoperative blood loss and length of hospital stay for the Endo-TLIF group (60.56 ± 0.36 mL, 8.12 ± 0.92 days, respectively) were statistically significantly lower than those for the MIS-TLIF group (65.47 ± 0.91 mL, 9.66 ± 1.34 days, respectively) (P < 0.05). The VAS and JOA scores of the patients in the two groups at postoperative 1 week, 3 months, 1 year, 2 years (Endo-TLIF VAS: 4.16 ± 0.92, 3.72 ± 1.54, 1.32 ± 0.45, 1.29 ± 0.34; JOA:16.71 ± 0.99, 19.86 ± 0.24, 24.91 ± 0.97, 25.88 ± 0.52; MIS-TLIF VAS: 4.17 ± 1.41, 2.98 ± 0.91, 1.54 ± 0.32, 1.33 ± 0.18; JOA: 16.67 ± 0.67, 19.58 ± 0.65, 25.33 ± 0.73, 25.69 ± 0.33) were statistically significantly improved from the preoperative scores (Endo-TLIF: 8.45 ± 1.44, 14.36 ± 0.56; MIS-TLIF: 8.11 ± 0.93, 14.45 ± 0.34, respectively) (P < 0.01). The VAS and JOA scores of the Endo-TLIF group were statistically significantly better than those of the MIS-TLIF group at 3 months and 1 year after surgery (P < 0.05). There were no statistically significant differences in the scores between the two groups at any of the other time points (P > 0.05). There was no significant difference in the intervertebral altitude between the two groups at the 3-month (11.36 ± 0.23, 11.21 ± 0.42, respectively) or final follow-up (10.88 ± 0.64, 10.81 ± 0.39, respectively) (P > 0.05). Dural tears, cerebrospinal fluid leakage, infection, and neurologic injury did not occur. Both groups showed good intervertebral fusion at the last follow-up. The intervertebral fusion rate was 97.5% (39/40) in the Endo-TLIF group and 94.7% (36/38) in the MIS-TLIF group, with no statistically significant difference between the two groups (χ2 = 0.118, P = 0.731). At the final follow-up, the modified MacNab's criteria were 92.5% and 89.5% between the two groups. CONCLUSION: Endo-TLIF with percutaneous pedicle screws (PPS) performed by using a visualization system for lumbar degenerative disease may be regarded as an efficient alternative surgery for degenerative lumbar spinal stenosis. It is a safe and minimally invasive way to perform this surgery and has shown satisfactory clinical outcomes.


Assuntos
Endoscopia/métodos , Degeneração do Disco Intervertebral/cirurgia , Vértebras Lombares/cirurgia , Parafusos Pediculares , Canal Medular/cirurgia , Fusão Vertebral/métodos , Estenose Espinal/cirurgia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Procedimentos Cirúrgicos Minimamente Invasivos/métodos , Medição da Dor , Estudos Retrospectivos , Inquéritos e Questionários
17.
Orthop Surg ; 13(2): 641-650, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33565271

RESUMO

To introduce a new surgery, percutaneous endoscopic unilateral laminotomy and bilateral decompression (Endo-ULBD) using visual trepan, and investigate its efficacy and safety in elderly patients with lumbar spinal stenosis. In our retrospective study, a total of 69 patients were enrolled between March 2018 and September 2018; 31 patients were treated with Endo-ULBD and 38 patients were treated with posterior lumbar interbody fusion surgery (PLIF). The operation time, intraoperative blood loss, and hospitalization duration were compared between the two groups. A visual analog scale (VAS) was used to evaluate the degree of pain. The Oswestry Disability Index (ODI) and European Quality of Life-5 Dimensions (EQ-5D) were used to evaluate lumbar function and quality of life, respectively. Lumbar X-ray, computed tomography (CT) and magnetic resonance imaging (MRI) were performed postoperatively at different time points. MacNab's outcome assessment and perioperative complications were also documented. The surgeon completed all surgeries successfully, and all 69 patients were followed up. The operative time of the Endo-ULBD group was 60.68 ± 0.47 min, while that of the PLIF group was 120.23 ± 10.24 min. The operative time of the Endo-ULBD group was shorter than that of the PLIF group, and the difference was statistically significant (P < 0.001). The volume of intraoperative blood loss was 47.25 ± 0.43 mL in the Endo-ULBD group and 256.90 ± 20.83 mL in the PILF group (P < 0.001). The length of hospital stay in the Endo-ULBD group was 5.12 ± 1.60 days and that in the PILF group was 10.54 ± 1.82 days (P < 0.001). The VAS scores at postoperative 1 day, 3 months, 6 months, final follow-up (Endo-ULBD: 6.58 ± 0.65, 4.55 ± 0.54, 2.78 ± 0.24, 1.31 ± 0.78; PLIF: 7.19 ± 1.14, 4.80 ± 0.13, 2.71 ± 0.83, 1.29 ± 0.56) were significantly improved compared with those before surgery (Endo-ULBD: 8.63 ± 0.37; PLIF: 8.31 ± 1.34). The ODI and EQ-5D scores of lumbar function and quality of life at each time point after surgery (Endo-ULBD ODI: 30.29% ± 0.47%, 23.35% ± 0.95%, 19.45% ± 0.81%, 10.84% ± 0.36%; EQ-5D: 0.38 ± 0.15, 0.45 ± 0.17, 0.63 ± 0.14, 0.71 ± 0.20; PLIF ODI: 33.56% ± 1.58%, 25.69% ± 2.69%, 20.01% ± 1.49%, 10.72% ± 0.29%; EQ-5D: 0.33 ± 0.03, 0.39 ± 0.05, 0.62 ± 0.07, 0.72 ± 0.10) were significantly improved compared with those before surgery (Endo-ULBD: 44.56 ± 1.32, 0.33 ± 0.07; PLIF: 43.79 ± 1.91, 0.31 ± 0.09, respectively), with statistically significant differences (P < 0.05); however, there was no significant difference between the two groups at the last follow-up (P > 0.05). At the last follow-up, the excellent and good efficacy rate was 90.3% (28/31) in the Endo-ULBD group and 89.4% (34/38) in the PILF group (χ2 = 0.089, P = 0.993). No mortality, irreversible nerve injury, or even paralysis occurred in either group. Endo-ULBD for lumbar spinal stenosis has the advantages of less trauma, a shortened operation time, and rapid recovery and is an effective alternative for the treatment of lumbar spinal stenosis. Strict surgical indications, reasonable surgical plans, and experienced surgeons are important factors to ensure safety and satisfactory postoperative efficacy.


Assuntos
Descompressão Cirúrgica/métodos , Endoscopia/métodos , Laminectomia/métodos , Vértebras Lombares/cirurgia , Fusão Vertebral/métodos , Estenose Espinal/cirurgia , Idoso , Descompressão Cirúrgica/instrumentação , Avaliação da Deficiência , Endoscopia/instrumentação , Feminino , Humanos , Masculino , Medição da Dor , Estudos Retrospectivos
18.
Orthop Surg ; 13(8): 2236-2245, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34668326

RESUMO

OBJECTIVE: To describe the rationale and application of triggered EMG (T-EMG) in intraoperative neurophysiological monitoring, and to explore the efficacy and safety of posterior percutaneous endoscopic cervical discectomy (PPECD) in the treatment of cervical spondylotic radiculopathy (CSR) under multimodal intraoperative neurophysiological monitoring (IOM). METHODS: This study was a retrospective cohort control study. The clinical data of 74 patients with single-segment CSR from June 2015 to August 2018 were analyzed retrospectively, of whom 35 underwent IOM-assisted PPECD with triggered EMG (T-EMG group), while 39 were subjected to IOM-assisted PPECD alone (IOM group). Operation time, hospital stay, and complications were recorded for both groups. The curative effect was evaluated according to the Visual Analog Scale (VAS) of neck and arm pain, Japanese Orthopaedic Association (JOA) score, and modified MacNab scale. RESULTS: Operations were successful and all patients were followed up for at least 24 (average 31.77 ± 9.51) months with no patient lost to follow-up. No significant difference was found in preoperative baseline data between the T-EMG and the IOM group (P > 0.05). Also, no significant difference was found in the operation time between the T-EMG (108.29 ± 11.44 min) and the IOM (110.13 ± 12.70 min) (P > 0.05) group, but the difference in hospital stay (T-EMG: 5.66 ± 0.99 days; IOM: 7.10 ± 1.43 days) was statistically significant (P < 0.05). The VAS for the neck and upper limbs in the two groups at 1 month post-operation (T-EMG: 2.09 ± 1.07, 2.26 ± 0.92; IOM:2.18 ± 1.05, 2.31 ± 0.77) and the last follow-up (T-EMG: 0.83 ± 0.62, 0.86 ± 0.55; IOM: 0.90 ± 0.50, 0.87 ± 0.61) were significantly different from the preoperative scores (T-EMG: 6.14 ± 1.09, 7.17 ± 1.04; IOM: 6.18 ± 1.28, 7.15 ± 1.23) (P < 0.05). However, no significant difference was found between the two groups (P > 0.05). The 1-month postoperative JOA scores for the two groups (12.69 ± 0.76; 12.59 ± 0.82) and those at the last follow-up (14.60 ± 0.77; 14.36 ± 0.78) were significantly different from the preoperative scores (11.09 ± 0.98; 11.05 ± 0.89) (P < 0.05), but the difference between the two groups was not significant (P > 0.05). One patient in the T-EMG group developed a transient aggravation of symptoms on the first day after surgery. In the IOM group, three patients had intraoperative cerebrospinal fluid leakage, and symptoms of C5 nerve root paralysis were presented in four patients following surgery. Compared with the IOM group, the T-EMG group had fewer complications (1/35; 7/39, P < 0.05). At the last follow-up, the modified MacNab criteria were 91.43% (32/35) and 89.7% (35/39) for the T-EMG group and IOM group, respectively. CONCLUSIONS: Triggered EMG prevents the occurrence of neurological complications, which not only aids PPECD for CSR treatment in achieving satisfactory results, but also reduces average hospital stay and complication rates.


Assuntos
Discotomia Percutânea/métodos , Eletromiografia/métodos , Endoscopia/métodos , Monitorização Neurofisiológica Intraoperatória/métodos , Radiculopatia/cirurgia , Espondilose/cirurgia , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Medição da Dor , Estudos Retrospectivos
19.
Zhongguo Gu Shang ; 34(12): 1103-7, 2021 Dec 25.
Artigo em Chinês | MEDLINE | ID: mdl-34965625

RESUMO

OBJECTIVE: To observe the curative effect of one-stage reconstruction of anterior cruciate ligament(ACL), posterior cruciate ligament (PCL) and medial collateral ligament (MCL) in patients with KD-Ⅲ-M knee injury, and to compare the operation time, hospitalization cost and curative effect after arthroscopic reconstruction of PCL with LARS artificial ligament and autogenous hamstring tendon, ACL reconstruction with autogenous hamstring tendon and MCL repair combined with limited incision. METHODS: From March 2016 to January 2019, a total of 36 patients met the criteria of this study. Twenty patients in group A were treated with autogenous hamstring tendon reconstruction of ACL and PCL and repair of MCL, including 17 males and 3 females, with an average age of (34.7±9.2) years old. Sixteen patients in group B with LARS artificial ligament reconstruction of PCL, with an autogenous hamstring tendon reconstruction of PCL and MCL repair as before as group B, including 15 males and 1 female, with an average age of (36.8±8.6) years old. The operation time, hospitalization time and total hospitalization cost were compared between the two groups. The preoperative and postoperative functions of the two groups were evaluated by Hospital for Sepcial Surgery (HSS) score and Lysholm score respectively, and the curative effects were compared within and between groups. RESULTS: All the patients in the two groups were followed up for at least 1 year. There were no complications such as infection and poor wound healing in both groups. There was significant difference in operation time between (120.25±9.55) min in group A and (106.63±8.85) min in group B (P<0.01). The average hospitalization days in group A was (10.60±1.64) days, while that in group B was (10.38±1.59) days. There was no significant difference between the two groups (P>0.05). The HSS score of group A increased from preoperative 32.95±5.03 to postoperative 84.70±5.72 (P<0.01), and that of group B increased from preoperative 33.81±4.10 to postoperative 85.00±5.25 (P<0.01). The Lyshlom score in group A increased from preoperative 21.10±3.46 to postoperative 80.25±5.75 (P<0.01), and in group B increased from preoperative 21.56±3.01 to postoperative 80.00±4.30(P<0.01). There was no significant difference in preoperative and postoperative scores between the two groups(P>0.05). CONCLUSION: There was no significant difference in the average hospitalization days between the two groups, but the operation time in group A was longerthan that in group B, and the hospitalization cost in group B was higher than that in group A. There was no difference in HSS score and Lysholm score before and follow-up for a certain period of time after operation.


Assuntos
Lesões do Ligamento Cruzado Anterior , Tendões dos Músculos Isquiotibiais , Luxação do Joelho , Reconstrução do Ligamento Cruzado Posterior , Ligamento Cruzado Posterior , Adulto , Ligamento Cruzado Anterior/cirurgia , Lesões do Ligamento Cruzado Anterior/cirurgia , Artroscopia , Feminino , Tendões dos Músculos Isquiotibiais/cirurgia , Humanos , Articulação do Joelho/cirurgia , Masculino , Pessoa de Meia-Idade , Ligamento Cruzado Posterior/cirurgia , Resultado do Tratamento
20.
Bone Joint Res ; 9(6): 322-332, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32670565

RESUMO

AIMS: The aim of this study was to examine whether tourniquet use can improve perioperative blood loss, early function recovery, and pain after primary total knee arthroplasty (TKA) in the setting of multiple-dose intravenous tranexamic acid. METHODS: This was a prospective, randomized clinical trial including 180 patients undergoing TKA with multiple doses of intravenous tranexamic acid. One group was treated with a tourniquet during the entire procedure, the second group received a tourniquet during cementing, and the third group did not receive a tourniquet. All patients received the same protocol of intravenous tranexamic acid (20 mg/kg) before skin incision, and three and six hours later (10 mg/kg). The primary outcome measure was perioperative blood loss. Secondary outcome measures were creatine kinase (CK), CRP, interleukin-6 (IL-6), visual analogue scale (VAS) pain score, limb swelling ratio, quadriceps strength, straight leg raising, range of motion (ROM), American Knee Society Score (KSS), and adverse events. RESULTS: The mean total blood loss was lowest in the no-tourniquet group at 867.32 ml (SD 201.11), increased in the limited-tourniquet group at 1024.35 ml (SD 176.35), and was highest in the tourniquet group at 1,213.00 ml (SD 211.48). The hidden blood loss was lowest in the no-tourniquet group (both p < 0.001). There was less mean intraoperative blood loss in the tourniquet group (77.48 ml (SD 24.82)) than in the limited-tourniquet group (137.04 ml (SD 26.96)) and the no-tourniquet group (212.99 ml (SD 56.35); both p < 0.001). Patients in the tourniquet group showed significantly higher levels of muscle damage and inflammation biomarkers such as CK, CRP, and IL-6 than the other two groups (p < 0.05). Outcomes for VAS pain scores, limb swelling ratio, quadriceps strength, straight leg raising, ROM, and KSS were significantly better in the no-tourniquet group at three weeks postoperatively (p < 0.05), but there were no significant differences at three months. No significant differences were observed among the three groups with respect to transfusion rate, thrombotic events, or the length of hospital stay. CONCLUSION: Patients who underwent TKA with multiple doses of intravenous tranexamic acid but without a tourniquet presented lower total blood loss and hidden blood loss, and they showed less postoperative inflammation reaction, less muscle damage, lower VAS pain score, and better early knee function. Our results argue for not using a tourniquet during TKA.Cite this article: Bone Joint Res 2020;9(6):322-332.

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