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Nontraumatic avascular necrosis of the femoral head(NANFH) is a common and refractory femoral head disease that causes bone death due to interruption of blood supply. Early clinical symptoms are atypical, such as hip pain and limited joint function. In the late stage, severe pain, shortening of the affected limb, claudication, and other serious symptoms are common, which se-riously affects the quality of life of patients. Therefore, it is of great significance to actively improve the clinical symptoms of NANFH to enhance the quality of life of patients. The pathogenesis of NANFH is complex, such as traumatic vascular circulatory disorders, the use of hormones or other drugs, alcoholism, and diabetes mellitus. These factors directly or indirectly lead to femoral head vascular damage, thrombosis, and coagulation system disorders, which reduce the blood supply to the acetabulum and femoral head, thus causing ischaemic death of the femoral head or even femoral head collapse. NANFH is mainly categorized as "bone impotence" and "bone paralysis" in traditional Chinese medicine(TCM). The treatment of NANFH with TCM has the characteristics and advantages of a long history, stable and reliable therapeutic effect, fewer adverse reactions, good patient tolerance, and high acceptance. Previous studies have shown that the promotion of angiogenesis is a key initiative in the prevention and treatment of NANFH, and TCM can promote fe-moral head angiogenesis by interfering with the expression of angiogenesis-related factors, which in turn can help to restore the blood supply of the femoral head and thus improve clinical symptoms of NANFH and prevent and treat NANFH. This article described the roles of blood supply interruption and angiogenesis in NANFH and the accumulated knowledge and experience of TCM in NANFH and summarized the role of angiogenesis-related factors in NANFH and the research progress on TCM intervention, so as to provide an idea for the subsequent research and a new basis for the clinical application of TCM in the treatment of NANFH.
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Medicamentos de Ervas Chinesas , Necrose da Cabeça do Fêmur , Humanos , Necrose da Cabeça do Fêmur/prevenção & controle , Necrose da Cabeça do Fêmur/tratamento farmacológico , Medicamentos de Ervas Chinesas/uso terapêutico , Medicamentos de Ervas Chinesas/administração & dosagem , Medicina Tradicional Chinesa , Animais , Cabeça do Fêmur/irrigação sanguínea , Neovascularização Patológica/tratamento farmacológico , Neovascularização Fisiológica/efeitos dos fármacos , AngiogêneseRESUMO
INTRODUCTION: Rheumatoid arthritis (RA) is a chronic inflammatory illness that mostly affects the joints of the hands and feet and can reduce life expectancy by an average of 3 to 10 years. Although tremendous progress has been achieved in the treatment of RA, a large minority of patients continue to respond poorly to existing medications, owing in part to a lack of appropriate therapeutic targets. METHODS: To find therapeutic targets for RA, a Mendelian randomization (MR) was performed. Cis-expression quantitative trait loci (cis-eQTL, exposure) data were obtained from the eQTLGen Consortium (sample size 31,684). Summary statistics for RA (outcome) were obtained from two largest independent cohorts: sample sizes of 97,173 (22,350 cases and 74,823 controls) and 269,377 (8279 cases and 261,098), respectively. Colocalisation analysis was used to test whether RA risk and gene expression were driven by common SNPs. Drug prediction and molecular docking was further used to validate the medicinal value of drug targets. RESULTS: Seven drug targets were significant in both cohorts in MR analysis and supported by localization. PheWAS at the gene level showed only ATP2A1 associated with other traits. These genes are strongly associated with immune function in terms of biological significance. Molecular docking showed excellent binding for drugs and proteins with available structural data. CONCLUSION: This study identifies seven potential drug targets for RA. Drugs designed to target these genes have a higher chance of success in clinical trials and is expected to help prioritise RA drug development and save on drug development costs.
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Artrite Reumatoide , Análise da Randomização Mendeliana , Humanos , Simulação de Acoplamento Molecular , Artrite Reumatoide/tratamento farmacológico , Artrite Reumatoide/genética , Sistemas de Liberação de Medicamentos , Desenvolvimento de MedicamentosRESUMO
Background: Interpersonal violence (IPV) against young women, including physical and sexual violence, poses a major threat to public health. We analyzed global, regional and national trends in violence against females aged 10-24 years from 1990 to 2019. Methods: We extracted age-standardized prevalence rates (ASPRs) of physical violence by firearm (PVF), physical violence by other means (PVOM), physical violence by sharp object (PVSO), and sexual violence (SV) from the Global Burden of Diseases, Injuries, and Risk Factors Study 2019. Joinpoint regression analysis calculated annual and average annual percentage changes (AAPCs) in ASPRs. Results: Globally, the ASPRs of the four measures of IPV decreased between 1990 and 2019, with the steepest declines between 2000 and 2009, except for SV, which increased slightly. However, the ASPRs of PVF and PVOM increased slightly between 2010 and 2019. Regionally, PVF prevalence declined most in East Asia (-0.9505, -1.0011 to -0.8975), South Asia (-0.277, -0.3089 to -0.244) and Latin America but PVOM prevalence increased in Oceania (0.6275, 0.6036 to 0.6498) and SV prevalence increased in Caribbean (0.4267, 0.4069 to 0.4495). Nationally, PVF prevalence decreased most in Thailand (-2.4031, -2.4634 to -2.3328) but increased most in Libya (6.8143, 6.6194 to 7.0113). SV prevalence increased most in Oman (0.4561, 0.4338 to 0.478) and the largest increase in Disability-adjusted life years (DALYs) from PVOM was observed in Botswana (6.2725, 6.0951 to 6.4082). DALYs showed similar trends. Conclusion: While global declines over 30 years are encouraging, IPV against young women persists. Urgent, tailored approaches across sectors are critical to curb drivers of violence against young women, including poverty, inequality and sociocultural attitudes. High-quality data and in-depth analyses can inform locally-relevant solutions. Overall, intensified political will and resource investment are needed to overcome this pervasive human rights violation.
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Osteoporosis is a serious public health problem that results in fragility fractures, especially in postmenopausal women. Because the current therapeutic strategy for osteoporosis has various side effects, a safer and more effective treatment is worth exploring. It is important to examine natural plant extracts during new drug design due to low toxicity. Mogrol is an aglycon of mogroside, which is the active component of Siraitia grosvenorii (Swingle) and exhibits anti-inflammatory, anticancer and neuroprotective effects. Here, we demonstrated that mogrol dose-dependently inhibited osteoclast formation and function. To confirm the mechanism, RNA sequencing (RNA-seq), real-time PCR (RT-PCR), immunofluorescence and Western blotting were performed. The RNA-seq data revealed that mogrol had an effect on genes involved in osteoclastogenesis. Furthermore, RT-PCR indicated that mogrol suppressed osteoclastogenesis-related gene expression, including CTSK, ACP5, MMP9 and DC-STAMP, in RANKL-induced bone marrow macrophages Western blotting demonstrated that mogrol suppressed osteoclast formation by blocking TNF receptor-associated factor 6 (TRAF6)-dependent activation of the mitogen-activated protein kinase nuclear factor-B (NF-κB) signaling pathway, which decreased two vital downstream transcription factors, the nuclear factor of activated T cells calcineurin-dependent 1 (NFATc1) and c-Fos proteins expression. Furthermore, mogrol dramatically reduced bone mass loss in postmenopausal mice. In conclusion, these data showed that mogrol may be a promising procedure for osteoporosis prevention or therapy.
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MiR-361-3p has been reported in several types of human cancer. However, the expression profile and biological functions of miR-361-3p in osteosarcoma remain uncovered. The expression profiles of miR-361-3p in osteosarcoma tissues and cell lines were evaluated using RT-qPCR. In 88 osteosarcoma patients, survival analysis was performed using Kaplan-Meier curves; while prognostic significance of miR-361-3p was analyzed using Cox regression analysis. The effects of miR-361-3p on cell proliferation, migration and invasion capacities were analyzed using CCK-8 and transwell assays. The target genes of miR-361-3p were assessed using luciferase reporter assay, RT-qPCR, Western blot and rescue experiments. Xenograft assay was conducted to test tumor growth ability. MiR-361-3p was found to be upregulated in human osteosarcoma tissues and cell lines. The expression of miR-361-3p was observed to be closely associated with TNM stage and lung metastasis. High expression of miR-361-3p was found to be capable of predicting poor clinical prognosis in osteosarcoma patients. Whilst overexpression of miR-361-3p was demonstrated to promote the proliferation, migration and invasion of osteosarcoma cells; knockdown of miR-361-3p was shown to exhibit an opposite inhibitory effect. Bioinformatics analysis and luciferase reporter assays confirmed that ARID3A is a direct target of miR-361-3p. Functional assays demonstrated that osteosarcoma cell proliferation, migration and invasion were promoted by miR-361-3p via negative regulation of ARID3A. Finally, overexpression of ARID3A was shown to partially reverse the tumor-promoting effect of miR-361-3p. Besides, in vivo assays revealed that miR-361-3p overexpression facilitated tumor growth in nude mice. In conclusion, this study indicates that miR-361-3p is a crucial prognostic biomarker of osteosarcoma, and that targeting of miR-361-3p/ARID3A axis may be a promising strategy in osteosarcoma therapy.
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Neoplasias Ósseas , Proteínas de Ligação a DNA , MicroRNAs , Osteossarcoma , Fatores de Transcrição , Animais , Neoplasias Ósseas/genética , Neoplasias Ósseas/metabolismo , Neoplasias Ósseas/patologia , Carcinogênese , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Xenoenxertos , Humanos , Camundongos , Camundongos Nus , MicroRNAs/genética , MicroRNAs/metabolismo , Osteossarcoma/genética , Osteossarcoma/metabolismo , Osteossarcoma/patologia , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismoRESUMO
Sensitive and reliable analysis of telomerase activity is important for clinical diagnosis, therapy, and prognosis of osteosarcoma. Telomerase activity is a complicated concept including both the amount of active telomerases and the length of the telomerases extension product. Still, few of the strategies formerly proposed distinguish the two aspects of telomerase activity. Herein, we propose a novel CRISPR-Cas12a-based fluorescent sensing platform that can output signals of both the amounts of telomerase and length of telomerase extension products with the assistance of an elegantly designed stem-loop probe and CRISPR-Cas12a system. On this basis, we induced a novel index, average telomerase activity, for accurate cancer reporting. Through systematic laboratory and clinical experiments, we have demonstrated that average telomerase activity can accurately distinguish cancer cells and has the potential for osteosarcoma staging.
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Neoplasias Ósseas/diagnóstico , Sistemas CRISPR-Cas/genética , Osteossarcoma/diagnóstico , Telomerase/metabolismo , Neoplasias Ósseas/metabolismo , Linhagem Celular Tumoral , Primers do DNA/metabolismo , Detecção Precoce de Câncer , Corantes Fluorescentes/química , Humanos , Técnicas de Amplificação de Ácido Nucleico , Osteossarcoma/metabolismo , RNA Guia de Cinetoplastídeos/metabolismoRESUMO
OBJECTIVE: Supracondylar humerus fractures are the most frequent fractures of the paediatric elbow. The present study introduced a modified surgical procedure for treatment of supracondylar humerus fractures in children. METHODS: From February 2015 to August 2019, 73 patients with Gartland's type II and III supracondylar fractures were treated with this modified method. Totally, 68 of all patients were followed up for 3-12 months (mean 8.25 months). The evaluation results included fracture nonunion, ulnar nerve injury, pin track infection, carrying angle and elbow joint Flynn score. RESULTS: The results showed that bone union was observed in all children, one case had an iatrogenic ulnar nerve injury, and the symptoms were completely relieved in 4 months after removing of the medial-side pin. All children had no cubitus varus deformity and no pin track infection, and the rate of satisfactory results according to Flynn's criteria score was 100%. CONCLUSION: The modified closed reduction and Kirschner wires internal fixation could effectively reduce the rate of open reduction, the risk of iatrogenic ulnar nerve injury, and the incidence of cubitus varus deformity in treatment of supracondylar humerus fractures in children.
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Fixação Interna de Fraturas/métodos , Fraturas do Úmero/cirurgia , Úmero/cirurgia , Procedimentos de Cirurgia Plástica , Fios Ortopédicos , Criança , Pré-Escolar , Feminino , Humanos , Fraturas do Úmero/fisiopatologia , Úmero/fisiopatologia , Masculino , PediatriaRESUMO
As a newly identified manganese transport protein, ZIP14 is highly expressed in the small intestine and liver, which are the two principal organs involved in regulating systemic manganese homeostasis. Loss of ZIP14 function leads to manganese overload in both humans and mice. Excess manganese in the body primarily affects the central nervous system, resulting in irreversible neurological disorders. Therefore, to prevent the onset of brain manganese accumulation becomes critical. In this study, we used Zip14-/- mice as a model for ZIP14 deficiency and discovered that these mice were born without manganese loading in the brain, but started to hyper-accumulate manganese within 3 weeks after birth. We demonstrated that decreasing manganese intake in Zip14-/- mice was effective in preventing manganese overload that typically occurs in these animals. Our results provide important insight into future studies that are targeted to reduce the onset of manganese accumulation associated with ZIP14 dysfunction in humans.
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Encéfalo/patologia , Proteínas de Transporte de Cátions/deficiência , Dieta , Suscetibilidade a Doenças , Manganês/metabolismo , Doenças Metabólicas/etiologia , Doenças Metabólicas/metabolismo , Animais , Encéfalo/metabolismo , Modelos Animais de Doenças , Fígado/metabolismo , Fígado/patologia , Manganês/efeitos adversos , Doenças Metabólicas/patologia , Doenças Metabólicas/prevenção & controle , Camundongos , Especificidade de ÓrgãosRESUMO
Neohesperidin has anti-oxidative and anti-inflammatory properties and exerts extensive therapeutic effects on various cancers. In this study, the osteosarcoma cell lines were exposed to different concentrations of neohesperidin. Cell proliferation and viability were assessed by CCK-8 and colony-formation assays. The role of neohesperidin in cell cycle progression and apoptosis were analyzed by flow cytometry and western blotting. To identify autophagosomes and autolysosomes, we used a tandem GFP-mRFP-LC3B lentiviral construct. In addition, autophagy was evaluated by examining autophagosome formation using transmission electron microscopy. Intracellular reactive oxygen species (ROS) production was detected by fluorescence microscopy and flow cytometry. Subsequently, the activation of the ROS/JNK signaling pathway was investigated. Neohesperidin could inhibit proliferation and induce apoptosis in SJSA and HOS cells. The formation of autophagosomes indicated that autophagy occurred in neohesperidin-treated cells and the apoptotic effect of neohesperidin was significantly increased after the use of autophagy inhibitors. Subsequently, we found that neohesperidin-induced apoptosis and autophagy were related to the increase in ROS generation and were significantly inhibited by GSH. Moreover, neohesperidin induced activation of the c-Jun N-terminal kinase (JNK) signaling pathway and inhibition of JNK with SP600125 attenuated neohesperidin-induced apoptosis and autophagy simultaneously. Our data indicated that neohesperidin caused G2/M phase arrest and induced apoptosis and autophagy by activating the ROS/JNK pathway in human osteosarcoma cells, suggesting that neohesperidin is a potential drug candidate for the treatment of osteosarcomas.
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Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Hesperidina/análogos & derivados , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Osteossarcoma/tratamento farmacológico , Espécies Reativas de Oxigênio/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Hesperidina/farmacologia , HumanosRESUMO
ZIP8 is a newly identified metal transporter. In human patients, mutations in ZIP8 result in severe manganese deficiency, suggesting a critical role for ZIP8 in regulating systemic manganese homeostasis. In mice, the deletion of ZIP8 recapitulates the symptoms of patients with ZIP8 mutations. However, further studies using mouse models to examine ZIP8's function were hindered by the lack of suitable antibodies to detect endogenous ZIP8 protein. In this study, we report the design, generation, and validation of a polyclonal antibody against mouse ZIP8. We have demonstrated that the newly generated antibody can be reliably used in immunoblotting analysis to detect endogenous ZIP8 protein in mouse tissues. The successful generation and validation of anti-mouse ZIP8 antibody provide opportunities to further examine the function and regulation of this metal transporter. In addition, our study may provide valuable insights into the future development of antibodies targeting polytopic membrane proteins.
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PURPOSE: The long-term survival rate of osteosarcoma, which is the most common type of primary malignant bone tumor, has stagnated in past decades. Acacetin is a natural flavonoid compound that has antioxidative and anti-inflammatory effects and exhibits extensive therapeutic effects on various cancers. In this study, the anticancer potential of acacetin and the underlying molecular mechanisms were examined in human osteosarcoma cells (SJSA and HOS). MATERIALS AND METHODS: HOS and SJSA cell lines were exposed to different concentrations of acacetin. Cell proliferation and viability were assessed by CCK-8 and colony-formation assays. Hoechst 33258 fluorescent staining was employed to detect apoptosis. Cell apoptosis was measured by an annexin V-FITC/PI assay by flow cytometry. The alteration in the mitochondrial membrane potential was detected by a JC-1 Assay Kit. Apoptosis-related protein expression was determined by Western blotting. Intracellular reactive oxygen species (ROS) production was detected by fluorescence microscopy and flow cytometry. Subsequently, the activation of the ROS/JNK signaling pathway was investigated. RESULTS: Acacetin could inhibit proliferation and induce apoptosis in SJSA and HOS cells. The acacetin treatment resulted in the activation of caspase-3, -8, and -9 and cleaved PARP. Further studies showed that acacetin-induced apoptosis was attributed to ROS. In addition, we found that acacetin induced the activation of the downstream c-Jun N-terminal kinase (JNK) signaling pathway. Subsequently, after treatment with the ROS scavenger GSH and the JNK inhibitor SP600125, the apoptosis-inducing effect triggered by acacetin was significantly attenuated. CONCLUSION: The results of the present study indicate that acacetin may induce apoptosis to inhibit cell growth by activating the ROS/JNK signaling pathway in SJSA and HOS cells, suggesting that acacetin may be a promising candidate for the management of osteosarcomas.
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Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Neoplasias Ósseas/tratamento farmacológico , Flavonas/farmacologia , Osteossarcoma/tratamento farmacológico , Antracenos/farmacologia , Neoplasias Ósseas/metabolismo , Neoplasias Ósseas/patologia , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Proteínas Quinases JNK Ativadas por Mitógeno/antagonistas & inibidores , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Osteossarcoma/metabolismo , Osteossarcoma/patologia , Espécies Reativas de Oxigênio/antagonistas & inibidores , Espécies Reativas de Oxigênio/metabolismoRESUMO
RATIONALE: We report a case of Schmorl node induced multiple radiculopathy. PATIENT CONCERNS: A 70-year-old female patient complained of lower back pain in the left leg accompanied by numbness and weakness. DIAGNOSIS: Radiographs showed obvious osteoporosis in the lumbar vertebrae. Computed tomography demonstrated a hole in the upper posterior half of the L2 vertebral body. Magnetic resonance imaging of the lumbar spine revealed a herniated disc involving a protrusion at the posterior wall of the L2 vertebral body, which was present in the left lateral and dorsal epidural spaces. There was significant lumbar stenosis at the L2 vertebral body secondary to dural sac compression due to the mass. INTERVENTION: Left-sided hemilaminectomy was performed at L2 with screw fixation at L1-3. Intraoperatively, the severely ruptured disc compression in the dural sac and nerve root was removed. OUTCOMES: The patient's leg pain was immediately resolved, and her back pain was reduced. The patient recovered normal motor function at 20 days after surgery. LESSONS: A Schmorl node can progress and break through the lumbar vertebral body, resulting in nerve compression. A large proximal herniated mass can cause distal multiple radiculopathy. Therefore, this special case of Schmorl node with multiple radiculopathy should be treated by removing the proximal herniated nucleus pulposus from the vertebral body.
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Deslocamento do Disco Intervertebral/complicações , Vértebras Lombares/patologia , Radiculopatia/etiologia , Idoso , Constrição Patológica , Feminino , Humanos , Deslocamento do Disco Intervertebral/cirurgia , Laminectomia/métodos , Dor Lombar , Vértebras Lombares/cirurgia , Osteoporose/complicaçõesRESUMO
Long noncoding RNAs (lncRNAs) have been certified as important regulators in tumorigenesis. LncRNA GAS6-AS2 (GAS6-AS2) was a newly identified tumor-related lncRNA, and its dysregulation and oncogenic effects in melanoma and bladder cancer had been reported in previous studies. However, the expression pattern and potential function of GAS6-AS2 in osteosarcoma (OS) have not been investigated. In this study, we identified a novel OS-related lncRNA GAS6-AS2. We found that GAS6-AS2 was distinctly upregulated in both OS specimens and cell lines. Distinct up-regulation of GAS6-AS2 in OS was correlated with advanced clinical stages and shorter survivals. In addition, USF1 could directly bind to the GAS6-AS2 promoter and contribute to its overexpression. Furthermore, GAS6-AS2 knockdown caused tumor suppressive effects via reducing cellular proliferation, migration and invasion, and promoting OS cell apoptosis. Besides, GAS6-AS2 directly bound to miR-934 and downregulated its expression. Mechanistically, GAS6-AS2 positively regulated the expression of BCAT1 through sponging miR-934. Taken together, our data illustrated how GAS6-AS2 played an oncogenic role in OS and might offer a potential therapeutic target for treating OS.
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Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , MicroRNAs/metabolismo , Osteossarcoma , RNA Longo não Codificante/metabolismo , Fatores Estimuladores Upstream/metabolismo , Carcinogênese/genética , Linhagem Celular Tumoral , Progressão da Doença , Regulação Neoplásica da Expressão Gênica , Humanos , Osteossarcoma/genética , Osteossarcoma/patologia , Transaminases/metabolismo , Ativação Transcricional , Regulação para CimaRESUMO
IGF-1R is expressed abnormally in osteosarcoma (OS) and could participate in its progression. In this study, we aimed to explore the effect of the IGF-1R inhibitor PQ401 as a treatment for OS. The relative expression of IGF-1R in OS patient tumors and the U2OS cell line were determined by qRT-PCR and by accessing information in a public database. Inhibition of cell proliferation by PQ401 was determined by MTT assay. Cell migration under low concentration treatment of PQ401 was carried out by transwell and wound healing assays. PQ401 induction of OS cell apoptosis was investigated by flow cytometry. Tumorigenesis under PQ401 treatment was evaluated by a colony formation assay. Finally, downstream blockage of the IGF-1R pathway was verified by western blotting. Our results show that the expression of IGF-1R was remarkably higher in OS cells, particularly in U2OS, than in other cancer-type cell lines. The inhibition of the IGF-1R pathway by PQ401 exhibited significant anticancer activity in the U2OS cell line in not only proliferation but also migration and colony formation. In addition, PQ401 is a strong inducer of OS cell apoptosis. Furthermore, western blotting was used to demonstrate that the IGF-1R related downstream pathway, including total ERK1/2, was significantly inhibited by PQ401. Thus, IGF-1R inhibition may represent a novel treatment for OS.
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Osteoporosis is a systemic bone metabolic disease that is highly prevalent in the elderly population, particularly in postmenopausal women, which results in enhanced bone fragility and an increased susceptibility to fractures. However, the underlying molecular pathogenesis mechanisms still remain to be further elucidated. In this study, in a rat ovariectomy- (OVX-) induced postmenopausal osteoporosis model, aberrant expression of a microRNA miR-142-5p and vascular cell adhesion molecule 1 (VCAM-1) was found by RNA sequencing analysis and qRT-PCR. Using a dual-luciferase reporter assay, we found that miR-142-5p can bind to and decrease expression of VCAM-1 mRNA. Such reduction was prohibited when the miR-142-5p binding site in VCAM-1 3'UTR was deleted, and Western blotting analyses validated the fact that miR-142-5p inhibited the expression of VCAM-1 protein. Bone marrow-derived mesenchymal stem cells (BMMSCs) transfected with miR-142-5p showed a significantly decreased migration ability in a Transwell migration assay. Collectively, these data indicated the important role of miR-142-5p in osteoporosis development involving targeting VCAM-1 and inhibiting BMMSC migration.
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Células da Medula Óssea/metabolismo , Movimento Celular , Regulação da Expressão Gênica , Células-Tronco Mesenquimais/metabolismo , MicroRNAs/metabolismo , Osteoporose/metabolismo , Molécula 1 de Adesão de Célula Vascular/biossíntese , Regiões 3' não Traduzidas , Animais , Células da Medula Óssea/patologia , Feminino , Células-Tronco Mesenquimais/patologia , MicroRNAs/genética , Osteoporose/genética , Osteoporose/patologia , Ratos , Ratos Sprague-Dawley , Molécula 1 de Adesão de Célula Vascular/genéticaRESUMO
BACKGROUND/AIMS: Transplantation of bone-marrow-derived mesenchymal stem cells (MSCs) promotes neural cell regeneration after spinal cord injury (SCI). Recently, we showed that suppression of microRNA-383 (miR-383) in MSCs increased the protein levels of glial cell line derived neurotrophic factor (GDNF), resulting in improved therapeutic effects on SCI. However, the overall effects of miR-383 suppression in MSCs on SCI therapy were not determined yet. Here, we addressed this question. METHODS: We used bioinformatics tools to predict all miR-383-targeting genes, confirmed the functional bindings in a dual luciferase reporter assay. The effects of alteration of candidate genes in MSCs on cell proliferation were analyzed by MTT assay and by Western blotting for PCNA. The effects on angiogenesis were assessed by HUVEC assay. The effects on SCI in vivo were analyzed by transplantation of the modified MSCs into nude rats that underwent SCI. RESULTS: Suppression of miR-383 in MSCs not only upregulated GDNF protein, but also increased vascular endothelial growth factor A (VEGF-A) and cyclin-dependent kinase 19 (CDK19), two other miR-383 targets. MiR-383-suppression-induced increases in CDK19 resulted in a slight but significant increase in MSC proliferation, while miR-383-suppression-induced increases in VEGF-A resulted in a slight but significant increase in MSC-mediated angiogenesis. CONCLUSIONS: Upregulation of CDK19 and VEGF-A by miR-383 suppression in MSCs further improve the therapeutic potential of MSCs in treating SCI in rats.
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Regulação da Expressão Gênica , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/metabolismo , MicroRNAs/genética , Traumatismos da Medula Espinal/terapia , Adulto , Animais , Proliferação de Células , Células Cultivadas , Regulação para Baixo , Fator Neurotrófico Derivado de Linhagem de Célula Glial/genética , Células HEK293 , Células Endoteliais da Veia Umbilical Humana , Humanos , Masculino , Transplante de Células-Tronco Mesenquimais/métodos , Células-Tronco Mesenquimais/citologia , Neovascularização Fisiológica , Ratos , Ratos Nus , Traumatismos da Medula Espinal/genética , Fator A de Crescimento do Endotélio Vascular/genéticaRESUMO
In general, the mammalian whole body mass-specific metabolic rate correlates positively with maximal urine concentration (Umax) irrespective of whether or not the species have adapted to arid or mesic habitat. Accordingly, we hypothesized that the thick ascending limb (TAL) of a rodent with markedly higher whole body mass-specific metabolism than rat exhibits a substantially higher TAL metabolic rate as estimated by Na+-K+-ATPase activity and Na+-K+-ATPase α1-gene and protein expression. The kangaroo rat inner stripe of the outer medulla exhibits significantly higher mean Na+-K+-ATPase activity (~70%) compared with two rat strains (Sprague-Dawley and Munich-Wistar), extending prior studies showing rat activity exceeds rabbit. Furthermore, higher expression of Na+-K+-ATPase α1-protein (~4- to 6-fold) and mRNA (~13-fold) and higher TAL mitochondrial volume density (~20%) occur in the kangaroo rat compared with both rat strains. Rat TAL Na+-K+-ATPase α1-protein expression is relatively unaffected by body hydration status or, shown previously, by dietary Na+, arguing against confounding effects from two unavoidably dissimilar diets: grain-based diet without water (kangaroo rat) or grain-based diet with water (rat). We conclude that higher TAL Na+-K+-ATPase activity contributes to relationships between whole body mass-specific metabolic rate and high Umax. More vigorous TAL Na+-K+-ATPase activity in kangaroo rat than rat may contribute to its steeper Na+ and urea axial concentration gradients, adding support to a revised model of the urine concentrating mechanism, which hypothesizes a leading role for vigorous active transport of NaCl, rather than countercurrent multiplication, in generating the outer medullary axial osmotic gradient.
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Peso Corporal , Metabolismo Energético , Capacidade de Concentração Renal , Medula Renal/enzimologia , Alça do Néfron/enzimologia , ATPase Trocadora de Sódio-Potássio/metabolismo , Sódio/urina , Animais , Dipodomys , Regulação Enzimológica da Expressão Gênica , Medula Renal/ultraestrutura , Alça do Néfron/ultraestrutura , Mitocôndrias/enzimologia , Osmorregulação , Ratos Sprague-Dawley , Ratos Wistar , Eliminação Renal , Reabsorção Renal , Especificidade da EspécieRESUMO
Understanding neurite outgrowth, orientation, and migration is important for the design of biomaterials that interface with the neural tissue. However, the molecular signaling alternations have not been well elucidated to explain the impact of hydrogels on cell morphology. In our previous studies, a silk fibroin peptide (SF16) hydrogel was found to be an effective matrix for the viability, morphology, and proliferation of PC12 rat pheocrhomocytoma cells. We found that PC12 cells in the peptide hydrogel exhibited adhesive morphology compared to those cultured in agarose or collagen. Moreover, we identified that cell adhesion molecules (E- and N-cadherin) controlled by mTOR signaling were highly induced in PC12 cells cultured in the SF16 peptide hydrogel. Our findings suggest that the SF16 peptide might be suitable to be a cell-adhesion material in cell culture or tissue engineering, and mTOR/cadherin signaling is required for the cell adhesion in the SF16-peptide hydrogel.
Assuntos
Caderinas/metabolismo , Adesão Celular , Proliferação de Células , Fibroínas/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Neurônios/enzimologia , Peptídeos/metabolismo , Serina-Treonina Quinases TOR/metabolismo , Alicerces Teciduais , Animais , Adesão Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Forma Celular , Hidrogéis , Neurônios/efeitos dos fármacos , Células PC12 , Inibidores de Proteínas Quinases/farmacologia , Ratos , Transdução de Sinais , Sirolimo/farmacologia , Serina-Treonina Quinases TOR/antagonistas & inibidoresRESUMO
Myeloid derived suppressor cells (MDSC) are very important in tumor immune evasion and they dramatically increased in peripheral blood of patients with osteosarcoma cancer. The association between MDSC and various cytokines has been studied in the peripheral blood. However, little is known about the mechanism drawing MDSC into tumor parenchyma. This study was to analyze the correlation between MDSC subsets and interleukin 18 (IL-18) level in osteosarcoma tumor model and its effect on the immunotherapy. MDSC were isolated from the blood and parenchyma and analyzed in the osteosarcoma tumor model. IL-18 levels were detected by enzyme-linked immunosorbent assay (ELISA) assay, real-time PCR, western blot and flow cytometry. Moreover, combination treatment with IL-18 inhibition and anti-PD1 was conducted to assess the therapeutic effects of IL-18 blockade. Results showed MDSC levels had a positive correlation with IL-18, suggesting IL-18 may attract MDSC into the parenchyma. IL-18 gene and protein expression significantly increased in blood and tumor lysates of tumor-bearing mice. Anti-IL-18 treatment significantly decreased G-MDSC and M-MDSC in the peripheral blood and tumor. Furthermore, combination therapy decreased the tumor burden and increased CD4+ and CD8+ T cell infiltration, as well as the production of interferon gamma (IFNγ) and granzyme B. Our study revealed a possible correlation between MDSC subsets and IL-18 inducing MDSC migration into the tumor tissue, in addition to provide the potential target to enhance the efficacy of immunotherapy in patients with osteosarcoma.
RESUMO
We report the development of a unique scalable Fourier transform 4-f system for instantly structured illumination in lithography. In the 4-f system, coupled with a 1-D grating and a phase retarder, the ±1st order of diffracted light from the grating serve as coherent incident sources for creating interference patterns on the image plane. By adjusting the grating and the phase retarder, the interference fringes with consecutive frequencies, as well as their orientations and phase shifts, can be generated instantly within a constant interference area. We demonstrate that by adapting this scalable Fourier transform system into lithography, the pixelated nano-fringe arrays with arbitrary frequencies and orientations can be dynamically produced in the photoresist with high variation resolution, suggesting its promising application for large-area functional materials based on space-variant nanostructures in lithography.