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INTRODUCTION: Acute anterior shoulder dislocation (AASD) is the most common joint dislocation. Here, we introduced a new reduction technique for AASD, named "Han's technique" (or "Touch overhead technique"). METHODS: Patients diagnosed with AASD were treated with "Han's technique" in the orthopaedic department of our hospital from October 2018 to November 2020. An orthopedic surgeon performed the reduction maneuver without any anesthesia or sedation throughout the reduction process. The fundamental information and related data were recorded, including patients' age, sex, dislocation side, previous dislocations history, reduction time, number of attempts at reduction, success rate of the reduction, intensity of pain during reduction using the 10-point visual analogue scale score (VAS), any complications, with or without the fracture and neurovascular examination results. RESULTS: Forty-one patients with AASD were involved in our study. Thirty-nine cases (95%) were primary dislocation. Eleven patients (27%) were complicated with large tubercle fracture and one patient (2%) complicated with inferior glenoid fractures. All patients were successfully reduced by Han's technique with mean reduction time was 138 s. The pain score during the reduction operation is only1.83 ± 0.83 points. No neurovascular injury or iatrogenic fracture was found after reduction in all patients. CONCLUSIONS: Han's technique (or Touch overhead technique) is a simple, safe, effective, mild and easy to master which can be operated by one surgeon without anesthesia or sedation for AASD.
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Anestesia , Luxação do Ombro , Fraturas do Ombro , Humanos , Luxação do Ombro/cirurgia , Luxação do Ombro/complicações , Manipulação Ortopédica/métodos , Fraturas do Ombro/cirurgia , DorRESUMO
Nerve damage can lead to movement and sensory dysfunction, with high morbidity and disability rates causing severe burdens on patients, families, and society. DNA methylation is a kind of epigenetics, and a great number of previous studies have demonstrated that DNA methylation plays an important role in the process of nerve regeneration and remodeling. However, compared with the central nervous system, the peripheral nervous system shows stronger recovery after injury, which is related to the complex microenvironment and epigenetic changes occurring at the site of injury. Therefore, what common epigenetic changes between the central and peripheral nervous systems remain to be elucidated. We first screened differential methylation genes after spinal cord injury and sciatic nerve injury using whole-genome bisulfite sequencing and methylated DNA immunoprecipitation sequencing, respectively. Subsequently, a total of 16 genes had the same epigenetic changes after spinal cord injury and sciatic nerve injury. The Gene Ontology analysis and Kyoto Encyclopedia of Genes and Genomes enrichment analysis were performed to identify the critical biological processes and pathways. Furthermore, a protein-protein interaction network analysis indicated that Dnm3, Ntrk3, Smurf1, Dpysl2, Kalrn, Shank1, Dlg2, Arsb, Reln, Bmp5, Numbl, Prickle2, Map6, and Htr7 were the core genes. These outcomes may provide novel insights into the molecular mechanism of the subacute phase of nerve injury. These verified genes can offer potential diagnostic and therapeutic targets for nerve injury.
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Metilação de DNA/genética , Traumatismos dos Nervos Periféricos/genética , Neuropatia Ciática/genética , Traumatismos da Medula Espinal/genética , Animais , Microambiente Celular/genética , Epigênese Genética/genética , Regulação da Expressão Gênica/genética , Genoma/genética , Humanos , Masculino , Traumatismos dos Nervos Periféricos/patologia , Mapas de Interação de Proteínas/genética , Ratos , Proteína Reelina , Nervo Isquiático/lesões , Nervo Isquiático/metabolismo , Nervo Isquiático/patologia , Neuropatia Ciática/patologia , Análise de Sequência de DNA , Traumatismos da Medula Espinal/patologiaRESUMO
BACKGROUND Spinal cord injury (SCI) is a serious disease with high disability and mortality rates, with no effective therapeutic strategies available. In SCI, abnormal DNA methylation is considered to be associated with axonal regeneration and cell proliferation. However, the roles of key genes in potential molecular mechanisms of SCI are not clear. MATERIAL AND METHODS Subacute spinal cord injury models were established in Wistar rats. Histological observations and motor function assessments were performed separately. Whole-genome bisulfite sequencing (WGBS) was used to detect the methylation of genes. Gene ontology (GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed using the DAVID database. Protein-protein interaction (PPI) networks were analyzed by Cytoscape software. RESULTS After SCI, many cavities, areas of necrotic tissue, and many inflammatory cells were observed, and motor function scores were low. After the whole-genome bisulfite sequencing, approximately 96 DMGs were screened, of which 50 were hypermethylated genes and 46 were hypomethylated genes. KEGG pathway analysis highlighted the Axon Guidance pathway, Endocytosis pathway, T cell receptor signaling pathway, and Hippo signaling pathway. Expression patterns of hypermethylated genes and hypomethylated genes detected by qRT-PCR were the opposite of WGBS data, and the difference was significant. CONCLUSIONS Abnormal methylated genes and key signaling pathways involved in spinal cord injury were identified through histological observation, behavioral assessment, and bioinformatics analysis. This research can serve as a source of additional information to expand understanding of spinal cord-induced epigenetic changes.
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Metilação de DNA , Traumatismos da Medula Espinal/genética , Animais , Biologia Computacional/métodos , Modelos Animais de Doenças , Epigênese Genética , Feminino , Perfilação da Expressão Gênica/métodos , Ontologia Genética , Redes Reguladoras de Genes , Mapas de Interação de Proteínas , Ratos , Ratos Wistar , Transdução de Sinais , Medula Espinal/patologia , Traumatismos da Medula Espinal/patologiaRESUMO
Pathological scars, such as keloids and hypertrophic scars, readily cause physical and psychological problems. Combination 5-fluorouracil (5-FU) with triamcinolone acetonide (TAC) is presumed to enhance the treatment of pathological scars, although supportive evidence is lacking. We aimed to compare the efficacy and safety of TAC alone and in combination with 5-FU for the treatment of hypertrophic scars and keloids. Five databases (PubMed, Medline, Cochrane databases, Embase and CNKI) were searched with the limitations of human subjects and English-language text. Mean differences (MDs), odds ratios (ORs) and 95% confidence intervals (CIs) were calculated. The Cochrane Collaboration's Risk of Bias Tool was used to assess the risk of bias. The control group received intralesional TAC alone, and the experimental group received TAC combined with 5-FU injection. A pooled analysis of the effectiveness based on patient self-assessment after treatment showed that the experimental group achieved better results than the control group (OR = 2·92, 95% CI = 1·63-5·22, P = 0·0003). Similarly, a pooled analysis of the effectiveness based on observer assessment following treatment produced the same conclusion (OR = 4·03, 95% CI = 1·40-11·61, P = 0·010). A meta-analysis of scar height after treatment showed that the experimental group performed better than the control group (MD = -0·14, 95% CI = -0·23-0·05, P = 0·002). The erythema score of the experimental group after treatment was superior (MD = -0·20, 95% CI = -0·34-0·06, P = 0·004). The heterogeneity test showed no heterogeneity among the studies (P > 0·1, I2 = 0%). TAC combined with 5-FU is more suitable for the treatment and prevention of hypertrophic scars and keloids, with greater improvement in scar height and patient satisfaction as well as fewer side effects.
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Anti-Inflamatórios/uso terapêutico , Cicatriz Hipertrófica/tratamento farmacológico , Fluoruracila/uso terapêutico , Queloide/tratamento farmacológico , Triancinolona Acetonida/uso terapêutico , Ferimentos e Lesões/tratamento farmacológico , Quimioterapia Combinada , Feminino , Humanos , Masculino , Resultado do TratamentoRESUMO
Cell death is an important part of the life cycle, serving as a foundation for both the orderly development and the maintenance of physiological equilibrium within organisms. This process is fundamental, as it eliminates senescent, impaired, or aberrant cells while also promoting tissue regeneration and immunological responses. A novel paradigm of programmed cell death, known as disulfidptosis, has recently emerged in the scientific circle. Disulfidptosis is defined as the accumulation of cystine by cancer cells with high expression of the solute carrier family 7 member 11 (SLC7A11) during glucose starvation. This accumulation causes extensive disulfide linkages between F-actins, resulting in their contraction and subsequent detachment from the cellular membrane, triggering cellular death. The RAC1-WRC axis is involved in this phenomenon. Disulfidptosis sparked growing interest due to its potential applications in a variety of pathologies, particularly oncology, neurodegenerative disorders, and metabolic anomalies. Nonetheless, the complexities of its regulatory pathways remain elusive, and its precise molecular targets have yet to be definitively identified. This manuscript aims to meticulously dissect the historical evolution, molecular underpinnings, regulatory frameworks, and potential implications of disulfidptosis in various disease contexts, illuminating its promise as a groundbreaking therapeutic pathway and target.
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Spinal cord injury (SCI) is a severe neurological disorder that causes neurological impairment and disability. Neural stem/progenitor cells (NS/PCs) derived from induced pluripotent stem cells (iPSCs) represent a promising cell therapy strategy for spinal cord regeneration and repair. However, iPSC-derived NS/PCs face many challenges and issues in SCI therapy; one of the most significant challenges is epigenetic regulation and that factors that influence this mechanism. Epigenetics refers to the regulation of gene expression and function by DNA methylation, histone modification, and chromatin structure without changing the DNA sequence. Previous research has shown that epigenetics plays a crucial role in the generation, differentiation, and transplantation of iPSCs, and can influence the quality, safety, and outcome of transplanted cells. In this study, we review the effects of epigenetic regulation and various influencing factors on the role of iPSC-derived NS/PCs in SCI therapy at multiple levels, including epigenetic reprogramming, regulation, and the adaptation of iPSCs during generation, differentiation, and transplantation, as well as the impact of other therapeutic tools (e.g., drugs, electrical stimulation, and scaffolds) on the epigenetic status of transplanted cells. We summarize our main findings and insights in this field and identify future challenges and directions that need to be addressed and explored.
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Células-Tronco Pluripotentes Induzidas , Células-Tronco Neurais , Traumatismos da Medula Espinal , Humanos , Epigênese Genética , Metilação de DNA , Células-Tronco Neurais/metabolismo , Células-Tronco Neurais/transplante , Traumatismos da Medula Espinal/genética , Traumatismos da Medula Espinal/terapia , Traumatismos da Medula Espinal/metabolismo , Diferenciação CelularRESUMO
Spinal cord injury (SCI) is a serious traumatic disease with no effective treatment. This study aimed to explore the therapeutic effect of syringaresinol on SCI. First, the potential targets and associated signaling pathways of syringaresinol were predicted by bioinformatics analysis and molecular docking. Second, MTT was employed to evaluate cell proliferation rate, Western blot was performed to detect protein expression, RT-qPCR was conducted to detect mRNA expression levels, flow cytometry and 5-ethynyl-2'-deoxyuridine (EDU) staining were used to determine cell apoptosis, and immunofluorescence and immunohistochemistry were used to estimate the expression of RNA binding fox-1 homolog 3 and clipped caspase 3. Basso-Beattie-Bresnahan scores and inclined plate tests were conducted to analyze hindlimb locomotor function. Results showed that syringaresinol could inhibit the apoptosis of glutamate-treated SHSY5Y cells by upregulating the expression of ubiquitination factor E4B (UBE4B) and activating the AKT serine/threonine kinase (AKT) signaling pathway. This effect can be rescued by UBE4B knockdown or AKT pathway inhibition. Syringaresinol remarkably improved locomotor function and increased neuronal survival in SCI rats. Our results suggested that syringaresinol could promote locomotor functional recovery by reducing neuronal apoptosis by activating the UBE4B/AKT signaling pathway.
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Proteínas Proto-Oncogênicas c-akt , Traumatismos da Medula Espinal , Ratos , Animais , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Medula Espinal/metabolismo , Ratos Sprague-Dawley , Simulação de Acoplamento Molecular , Transdução de Sinais , Traumatismos da Medula Espinal/metabolismo , Apoptose , Neurônios/metabolismo , Ubiquitinação , Serina/metabolismo , Recuperação de Função FisiológicaRESUMO
BACKGROUND: Gout is a prevalent manifestation of metabolic osteoarthritis induced by elevated blood uric acid levels. The purpose of this study was to investigate the mechanisms of gene expression regulation in gout disease and elucidate its pathogenesis. METHODS: The study integrated gout genome-wide association study (GWAS) data, single-cell transcriptomics (scRNA-seq), expression quantitative trait loci (eQTL), and methylation quantitative trait loci (mQTL) data for analysis, and utilized two-sample Mendelian randomization study to comprehend the causal relationship between proteins and gout. RESULTS: We identified 17 association signals for gout at unique genetic loci, including four genes related by protein-protein interaction network (PPI) analysis: TRIM46, THBS3, MTX1, and KRTCAP2. Additionally, we discerned 22 methylation sites in relation to gout. The study also found that genes such as TRIM46, MAP3K11, KRTCAP2, and TM7SF2 could potentially elevate the risk of gout. Through a Mendelian randomization (MR) analysis, we identified three proteins causally associated with gout: ADH1B, BMP1, and HIST1H3A. CONCLUSION: According to our findings, gout is linked with the expression and function of particular genes and proteins. These genes and proteins have the potential to function as novel diagnostic and therapeutic targets for gout. These discoveries shed new light on the pathological mechanisms of gout and clear the way for future research on this condition.
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Predisposição Genética para Doença , Estudo de Associação Genômica Ampla , Gota , Análise da Randomização Mendeliana , Locos de Características Quantitativas , Análise de Célula Única , Gota/genética , Humanos , Análise da Randomização Mendeliana/métodos , Estudo de Associação Genômica Ampla/métodos , Predisposição Genética para Doença/genética , Locos de Características Quantitativas/genética , Análise de Célula Única/métodos , Metilação de DNA/genética , Polimorfismo de Nucleotídeo Único , Mapas de Interação de Proteínas/genética , Álcool DesidrogenaseRESUMO
BACKGROUND: Post-traumatic related limb osteomyelitis (PTRLO) is a complex bone infection. Currently, there are no available microbial data on a national scale that can guide appropriate antibiotic selection, and explore the dynamic changes in dominant pathogens over time. This study aimed to conduct a comprehensive epidemiological analysis of PTRLO in China. METHODS: The study was approved by the Institutional Research Board (IRB), and 3526 PTRLO patients were identified from 212 394 traumatic limb fracture patients at 21 hospitals between 1 January 2008 and 31 December 2017. A retrospective analysis was conducted to investigate the epidemiology of PTRLO, including changes in infection rate (IR), pathogens, infection risk factors and antibiotic resistance and sensitivity. RESULTS: The IR of PTRLO increased gradually from 0.93 to 2.16% (Z=14.392, P <0.001). Monomicrobial infection (82.6%) was significantly higher than polymicrobial infection (17.4%) ( P <0.001). The IR of Gram-positive (GP) and Gram-negative (GN) pathogens showed a significant increase from the lowest 0.41% to the highest 1.15% (GP) or 1.62% (GN), respectively. However, the longitudinal trend of GP vs. GN's composition did not show any significance (Z=±1.1918, P >0.05). The most prevalent GP strains were Methicillin-sensitive Staphylococcus aureus (MSSA) (17.03%), Methicillin-resistant Staphylococcus aureus (MRSA) (10.46%), E. faecalis (5.19%) and S. epidermidis (4.87%). In contrast, the dominant strains GN strains were Pseudomonas Aeruginosa (10.92%), E. cloacae (10.34%), E. coli (9.47%), Acinetobacter Baumannii (7.92%) and Klebsiella Pneumoniae (3.33%). In general, the high-risk factors for polymicrobial infection include opened-fracture (odds ratio, 2.223), hypoproteinemia (odds ratio, 2.328), and multiple fractures (odds ratio, 1.465). It is important to note that the antibiotics resistance and sensitivity analysis of the pathogens may be influenced by complications or comorbidities. CONCLUSIONS: This study provides the latest data of PTRLO in China and offers trustworthy guidelines for clinical practice. (China Clinical Trials.gov number, ChiCTR1800017597).
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Coinfecção , Fraturas Expostas , Staphylococcus aureus Resistente à Meticilina , Osteomielite , Humanos , Estudos Retrospectivos , Escherichia coli , Coinfecção/tratamento farmacológico , Testes de Sensibilidade Microbiana , Antibacterianos/uso terapêutico , China/epidemiologia , Osteomielite/epidemiologia , Osteomielite/etiologia , Osteomielite/tratamento farmacológicoRESUMO
N6-methyladenosine (m6A), an essential post-transcriptional modification in eukaryotes, is closely related to the development of pathological processes in neurological diseases. Notably, spinal cord injury (SCI) is a serious traumatic disease of the central nervous system, with a complex pathological mechanism which is still not completely understood. Recent studies have found that m6A modification levels are changed after SCI, and m6A-related regulators are involved in the changes of the local spinal cord microenvironment after injury. However, research on the role of m6A modification in SCI is still in the early stages. This review discusses the latest progress in the dynamic regulation of m6A modification, including methyltransferases ("writers"), demethylases ("erasers") and m6A -binding proteins ("readers"). And then analyses the pathological mechanism relationship between m6A and the microenvironment after SCI. The biological processes involved included cell death, axon regeneration, and scar formation, which provides new insight for future research on the role of m6A modification in SCI and the clinical transformation of strategies for promoting recovery of spinal cord function.
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Osteoporosis (OP) is an aging-related disease that is the main etiology of fragility fracture. Qing'e Pill (QEP) is a mixture of traditional Chinese medicine (TCM) consisting of Eucommia ulmoides Oliv., Psoralea corylifolia L., Juglans regia L., and Allium sativum L. QEP has an anti-osteoporosis function, but the underlying mechanism remains unclear. In this study, online databases were employed to determine the chemical compounds of QEP and potential target genes in osteoporosis. Potential pathways associated with genes were defined by Gene Ontology (GO) and Kyoto Encyclopaedia of Genes and Genomes (KEGG) databases. A compound-target-disease network was constructed. Hub genes screened through Cytoscape were intersected with the FerrDB database. The potential key genes were validated in HFOB 1.19 cells, and rat models were ovariectomized through Western blot, RT-qPCR, ELISA, HE staining, immunohistochemistry, and immunofluorescence analyses. The intersection targets of QEP and osteoporosis contained 121 proteins, whereas the target-pathway network included 156 pathways. We filtered five genes that stood out in the network analysis for experimental verification. The experiments validated that QEP exerted therapeutic effects on osteoporosis by inhibiting ferroptosis and promoting cell survival via the PI3K/AKT pathway and ATM. In conclusion, combining the application of network analysis and experimental verification may provide an efficient method to validate the molecular mechanism of QEP on osteoporosis.
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Vitamin C deficiency disrupts the integrity of connective tissues including bone. For decades this function has been primarily attributed to Vitamin C as a cofactor for collagen maturation. Here, we demonstrate that Vitamin C epigenetically orchestrates osteogenic differentiation and function by modulating chromatin accessibility and priming transcriptional activity. Vitamin C regulates histone demethylation (H3K9me3 and H3K27me3) and promotes TET-mediated 5hmC DNA hydroxymethylation at promoters, enhancers and super-enhancers near bone-specific genes. This epigenetic circuit licenses osteoblastogenesis by permitting the expression of all major pro-osteogenic genes. Osteogenic cell differentiation is strictly and continuously dependent on Vitamin C, whereas Vitamin C is dispensable for adipogenesis. Importantly, deletion of 5hmC-writers, Tet1 and Tet2, in Vitamin C-sufficient murine bone causes severe skeletal defects which mimic bone phenotypes of Vitamin C-insufficient Gulo knockout mice, a model of Vitamin C deficiency and scurvy. Thus, Vitamin C's epigenetic functions are central to osteoblastogenesis and bone formation and may be leveraged to prevent common bone-degenerating conditions.
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Deficiência de Ácido Ascórbico , Osteogênese , Animais , Ácido Ascórbico/farmacologia , Deficiência de Ácido Ascórbico/genética , Calcificação Fisiológica/genética , Diferenciação Celular/genética , Cromatina , DNA/metabolismo , Metilação de DNA , Histonas/metabolismo , Camundongos , Osteogênese/genéticaRESUMO
OBJECTIVE: To explore the clinical characteristics and risk factors of refracture in patients suffering from osteoporosis-related fractures as well as effective interventions. METHODS: From January 2006 to January 2008, both out-patients and in-patients in our hospital who were over 50 years old and suffered from osteoporosis-related fractures were selected for this research. They were divided into fracture group and refracture group. The refracture rate was followed up for 2 years, during which 11 patients developed refracture, thus were included in the refracture group. Therefore, 273 patients, 225 first-fracture cases, aged (67.7+/-8.5) years, and 48 refracture cases, aged (72.7+/-9.5) years, were included in this study. General data including age and sex, fracture types, femoral neck bone mineral density (BMD) T-scores tested by dual-energy X-rays absorptiometry (DEXA), Charlson index, time-frame between two fractures as well as mobility skill assessment were collected and analyzed by single-factor and multivariate statistical methods. RESULTS: Females accounted for 70.2% of the fracture group and 77.1% of the refracture group. The most common refracture type was vertebral fracture for the first time and femoral neck fracture for the second time during the follow-up. The second fracture happened 3.7 years after the first one on average. The refracture rate was 2.12% within one year, and 4.66% within two years. Risk factors for a second fracture in osteoporotic fracture patients included age (larger than 75 years, HR equal to 1.23, 95%CI 1.18-1.29; larger than 85 years, HR equal to 1.68, 95% CI 1.60-1.76), female sex (HR equal to 1.36, 95%CI 1.32-1.40), prior vertebral fractures (HR equal to 1.62, 95%CI 1.01-2.07), prior hip fractures (HR equal to 1.27, 95%CI 0.89-2.42), BMD T-score less than -3.5 (HR equal to 1.38, 95%CI 1.17-1.72) and weakened motor skills (HR equal to 1.27, 95%CI 1.09-1.40). CONCLUSIONS: The risks of second fracture among patients with initial brittle fracture are substantial. There is adequate time between the first and second fractures for interventions to reduce the risks of refracture, especially for the old women with a vertebral or hip fracture. Medication, motor functional rehabilitation and fall-down prevention training are helpful.
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Fraturas por Osteoporose/etiologia , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Densidade Óssea , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Destreza Motora , Análise Multivariada , Fatores de RiscoRESUMO
BACKGROUND: Clearance of myelin debris and remyelination of myelin are necessary steps for peripheral nerve remodeling and regeneration. It has yet to be clarified which genes or proteins are involved in endocytosis or exocytosis in the removal of myelin debris during peripheral nerve repair. METHODS: For this project, a rat model of subacute stage of sciatic nerve injury was established first. Subsequently, normal Schwann cells (NSCs) and activated Schwann cells (ASCs) were harvest before and after peripheral nerve injury (PNI). Following methylated DNA immunoprecipitation sequencing (MeDIP-seq) and tandem mass tags (TMT) labeling analysis of NSCs and ASCs, what common biomarkers changes in peripheral nervous systems remain to be elucidated. RESULTS: A total of 14,770 different expression genes (DEGs) and 3249 different expression proteins (DEPs) were screened between ASCs and NSCs. For the exosomes, the diameter and particles concentration of exosomes were 141.7 ânm and 2.97 â× â107 particles/mL, respectively. The size distribution of exosomes was 50-200 ânm. ASCs showed higher cellular uptake ability than the NSCs by cellular uptake test. Moreover, RAB7A, ARF6, ARF1, VPS45, RAB11A, DNM3, and NEDD4 were the core markers and may control the molecular mechanism of the Endocytosis pathway. CONCLUSION: These biomarkers may play significant roles in the initiation phase of demyelination and axon regeneration. THE TRANSLATIONAL POTENTIAL OF THIS ARTICLE: This study explores that the endocytosis-associated patterns of Schwann cells may be new therapeutic strategy for nerve tissue engineering and nerve regeneration.
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N6-methyladenosine (m6A) is one of the most plentiful internal RNA modifications, especially in eukaryotic messenger RNA (mRNA), which plays pivotal roles in the regulation of mRNA life cycle and nerve development. However, the mRNA m6A methylation pattern in peripheral nervous injury (PNI) has not been investigated. In this study, sciatic nerve samples were collected from 7 days after sciatic nerve injury (SNI) and control rats. Quantitative real-time PCR demonstrated that m6A-related methyltransferase/demethylase genes were remarkably upregulated in SNI group compared with control group. Methylated RNA immunoprecipitation sequencing (MeRIP-seq) was performed to reveal the m6A methylation landscape. The results showed that 4,014 m6A peaks were significantly altered, including 2,144 upregulated and 1,870 downregulated m6A peaks, which were corresponded to 1,858 genes. Moreover, 919 differentially expressed genes were identified by the conjoint analysis of MeRIP-seq and RNA-seq. GO and KEGG pathway analyses were performed to determine the biological functions and signaling pathways of the m6A-modified genes. Notably, these genes were mainly related to the immune system process, cell activation, and nervous system development in GO analysis. KEGG pathway analysis revealed that these genes were involved in the cell cycle, B cell receptor signaling pathway, axon guidance pathway, and calcium signaling pathway. Furthermore, the m6A methylation and protein expression levels of autophagy-related gene (Atg7) were increased, together with the activation of autophagy. These findings shed some light on the epigenetic regulation of gene expression, which may provide a new opinion to promote functional recovery after PNI.
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Schwann cells are unique glial cells in the peripheral nervous system. These cells provide a range of cytokines and nutritional factors to maintain axons and support axonal regeneration. However, little is known concerning adhesion-associated epigenetic changes that occur in Schwann cells after peripheral nerve injury (PNI). In the present study, adhesion-associated DNA methylation biomarkers were assessed between normal and injury peripheral nerve. Specifically, normal Schwann cells (NSCs) and activated Schwann cells (ASCs) were obtained from adult Wistar rats. After the Schwann cells were identified, proliferation and adhesion assays were used to assess differences between NSCs and ASCs. Methylated DNA immunoprecipitation-sequencing and bioinformatics analysis were used to identify and analyze the differentially methylated genes. Reverse transcription-quantitative PCR was performed to assess the expression levels of adhesion-associated genes. In the present study, the proliferation and adhesion assays demonstrated that ASCs had a more robust proliferative activity and adhesion compared with NSCs. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were performed to identify methylation-associated biological processes and signaling pathways. Protein-protein interaction network analysis revealed that Fyn, Efna1, Jak2, Vav3, Flt4, Epha7, Crk, Kitlg, Ctnnb1 and Ptpn11 were potential markers for Schwann cell adhesion. The expression levels of several adhesion-associated genes, such as vinculin, BCAR1 scaffold protein, collagen type XVIII α1 chain and integrin subunit ß6, in ASCs were altered compared with those in NSCs. The current study analyzed adhesion-associated DNA methylation patterns of Schwann cells and identified candidate genes that may potentially regulate Schwann cell adhesion in Wistar rats before and after PNI.
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OBJECTIVES: Schwann cells (SCs) have a wide range of applications as seed cells in the treatment of nerve injury during transplantation. However, there has been no report yet on kinds of proteomics changes that occur in Schwann cells before and after peripheral nerve injury. MATERIALS AND METHODS: Activated Schwann cells (ASCs) and normal Schwann cells (NSCs) were obtained from adult Wistar rat sciatic nerves. After immunofluorescence identification, we identified differentially expressed proteins in the ASCs and NSCs using isobaric tags for relative and absolute quantitation (iTRAQ) combined with high-resolution Orbitrap liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS). In addition, all the differentially expressed proteins were analyzed by Gene ontology (GO) analysis and Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis. Finally, several differentially expressed proteins were selected for Western blot verification. RESULTS: A total of 122 differentially expressed proteins in ASCs and NSCs were screened. GO analysis suggested that these different proteins are likely to accumulate in the cytoplasm and are associated with single-multicellular organism processes. The KEGG pathway analysis suggested that proteins related to purine metabolism were significantly enriched. The expression of Transmembrane glycoprotein NMB (GPNMB), Ectonucleotide pyrophosphatase/phosphodiesterase family member 3 (ENPP3), and other proteins were consistent with the proteomics data obtained by Western blot analysis. CONCLUSION: GPNMB, ENPP3, GFPT2, and other proteins may play an important role in the repair of peripheral nerve injury. This study may provide new insights into changes in SCs after peripheral nerve injury.
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BACKGROUND: Spinal cord injury (SCI) is a traumatic disease of the central nervous system, accompanied with high incidence and high disability rate. Tissue engineering scaffold can be used as therapeutic systems to provide effective repair for SCI. PURPOSE: In this study, a novel tissue engineering scaffold has been synthesized in order to explore the effect of nerve repair on SCI. PATIENTS AND METHODS: Polycaprolactone (PCL) scaffolds loaded with actived Schwann cells (ASCs) and induced pluripotent stem cells -derived neural stem cells (iPSC-NSCs), a combined cell transplantation strategy, were prepared and characterized. The cell-loaded PCL scaffolds were further utilized for the treatment of SCI in vivo. Histological observation, behavioral evaluation, Western-blot and qRT-PCR were used to investigate the nerve repair of Wistar rats after scaffold transplantation. RESULTS: The iPSCs displayed similar characteristics to embryonic stem cells and were efficiently differentiated into neural stem cells in vitro. The obtained PCL scaffolds werê0.5 mm in thickness with biocompatibility and biodegradability. SEM results indicated that the ASCs and (or) iPS-NSCs grew well on PCL scaffolds. Moreover, transplantation reduced the volume of lesion cavity and improved locomotor recovery of rats. In addition, the degree of spinal cord recovery and remodeling maybe closely related to nerve growth factor and glial cell-derived neurotrophic factor. In summary, our results demonstrated that tissue engineering scaffold treatment could increase tissue remodeling and could promote motor function recovery in a transection SCI model. CONCLUSION: This study provides preliminary evidence for using tissue engineering scaffold as a clinically viable treatment for SCI in the future.
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Células-Tronco Pluripotentes Induzidas/citologia , Células-Tronco Neurais/citologia , Poliésteres/química , Células de Schwann/citologia , Traumatismos da Medula Espinal/terapia , Engenharia Tecidual/métodos , Alicerces Teciduais/química , Animais , Axônios/patologia , Comportamento Animal , Separação Celular , Técnicas de Cocultura , Sangue Fetal/citologia , Células-Tronco Pluripotentes Induzidas/ultraestrutura , Leucócitos Mononucleares/citologia , Camundongos , Fatores de Crescimento Neural/metabolismo , Células-Tronco Neurais/metabolismo , Células-Tronco Neurais/ultraestrutura , Ratos Wistar , Células de Schwann/ultraestrutura , Medula Espinal/patologiaRESUMO
Spinal cord injury (SCI), for which there currently is no cure, is a heavy burden on patient physiology and psychology. The microenvironment of the injured spinal cord is complicated. According to our previous work and the advancements in SCI research, 'microenvironment imbalance' is the main cause of the poor regeneration and recovery of SCI. Microenvironment imbalance is defined as an increase in inhibitory factors and decrease in promoting factors for tissues, cells and molecules at different times and spaces. There are imbalance of hemorrhage and ischemia, glial scar formation, demyelination and re-myelination at the tissue's level. The cellular level imbalance involves an imbalance in the differentiation of endogenous stem cells and the transformation phenotypes of microglia and macrophages. The molecular level includes an imbalance of neurotrophic factors and their pro-peptides, cytokines, and chemokines. The imbalanced microenvironment of the spinal cord impairs regeneration and functional recovery. This review will aid in the understanding of the pathological processes involved in and the development of comprehensive treatments for SCI.
Assuntos
Traumatismos da Medula Espinal/patologia , Medula Espinal/patologia , Animais , Citocinas/análise , Hemorragia/etiologia , Humanos , Microglia/patologia , Fatores de Crescimento Neural/análise , Regeneração Nervosa , Células-Tronco Neurais/patologia , Neurônios/patologia , Medula Espinal/fisiopatologia , Traumatismos da Medula Espinal/complicações , Traumatismos da Medula Espinal/fisiopatologia , Traumatismos da Medula Espinal/terapiaRESUMO
This study aims to find the difference of genomewide DNA methylation in Schwann cells (SCs) before and after peripheral nerve system (PNS) injury by Methylated DNA Immunoprecipitation Sequencing (MeDIP-Seq) and seek meaningful differentially methylated genes related to repairment of injured PNS. SCs harvested from sciatic nerve were named as activated Schwann cells (ASCs), and the ones harvested from brachial plexus were named as normal Schwann cells (NSCs). Genomic DNA of ASCs and NSCs were isolated and MeDIP-Seq was conducted. Differentially methylated genes and regions were discovered and analyzed by bioinformatic methods. MeDIP-Seq analysis showed methylation differences were identified between ASCs and NSCs. The distribution of differentially methylated regions (DMRs) peaks in different components of genome was mainly located in distal intergenic regions. GO and KEGG analysis of these methylated genes were also conducted. The expression patterns of hypermethylated genes (Dgcr8, Zeb2, Dixdc1, Sox2, and Shh) and hypomethylated genes (Gpr126, Birc2) detected by qRT-PCR were opposite to the MeDIP analysis data with significance (p < 0.05), which proved MeDIP analysis data were real and believable. Our data serve as a basis for understanding the injury-induced epigenetic changes in SCs and the foundation for further studies on repair of PNS injury.