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Asthma is an inflammatory disease. Airway epithelial cell pyroptosis and cytokine secretion promote asthma progression. Tripartite motif 47 (TRIM47) belongs to the E3 ubiquitin ligase family and is associated with apoptosis and inflammation in a range of diseases. However, the role of TRIM47 in asthma has not been explored. In this study, the human bronchial epithelial cell line BEAS-2B was treated with house dust mite (HDM) and TRIM47 expression was detected by RT-qPCR and Western blot. After transfection with TRIM47 interfering and overexpressing plasmids, the synthesis and secretion of cytokines, as well as pyroptosis-related indicators, were examined. Nuclear factor kappa-B (NF-κB) pathway proteins and nod-like receptor protein 3 (NLRP3) inflammasome were measured to explore the mechanism of TRIM47 action. In addition, the effect of TRIM47 on the level of NF-κB essential modulator (NEMO) ubiquitination was detected by an immunoprecipitation assay. The results showed that TRIM47 was upregulated in HDM-induced BEAS-2B cells and that TRIM47 mediated HDM-induced BEAS-2B cell pyroptosis and cytokine secretion. Mechanistically, TRIM47 promoted the K63-linked ubiquitination of NEMO and facilitated NF-κB/NLRP3 pathway activation. In conclusion, TRIM47 may promote cytokine secretion mediating inflammation and pyroptosis in bronchial epithelial cells by activating the NF-κB/NLRP3 pathway. Therefore, TRIM47 may be a potential therapeutic target for HDM-induced asthma.
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Brônquios , Células Epiteliais , NF-kappa B , Proteína 3 que Contém Domínio de Pirina da Família NLR , Piroptose , Transdução de Sinais , Ubiquitinação , Humanos , NF-kappa B/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Células Epiteliais/metabolismo , Brônquios/metabolismo , Brônquios/patologia , Animais , Linhagem Celular , Quinase I-kappa B/metabolismo , Pyroglyphidae , Asma/metabolismo , Asma/patologia , Inflamassomos/metabolismo , Ubiquitina-Proteína Ligases/metabolismo , Ubiquitina-Proteína Ligases/genética , Citocinas/metabolismo , Proteínas com Motivo Tripartido/metabolismo , Proteínas com Motivo Tripartido/genéticaRESUMO
INTRODUCTION: Anaphylaxis is a serious systemic hypersensitivity reaction that usually has a rapid onset and may cause death. The aim of this study was to clarify the clinical characteristics of anaphylaxis in children of all ages in Xi'an, China. METHODS: A retrospective study was conducted on anaphylaxis cases in the emergency department of the Affiliated Children's Hospital of Xi'an Jiaotong University between January 1, 2016, and July 1, 2021. The statistical methods used were the χ2 test, Fisher's exact test, and Z-test. RESULTS: A total of 110 cases of anaphylaxis were collected: 70% were male, 13 were <1 year old, 17 were 1-2 years old, 42 were 3-6 years old, 38 were 7-16 years old, 10 (9.1%) had ≥2 anaphylaxis, and 75 (68.1%) had a previous history of allergy. The triggers of anaphylaxis were analyzed: 50 cases (45.5%) were induced by food, 37 cases (33.6%) by drugs, 6 cases (5.5%) by insect bites, 4 cases (3.6%) by exercise, and 12 cases (11.8%) by unknown causes. Common food allergens were milk (20%, 10/50), buckwheat (16%, 8/50), eggs (14%, 7/50), and fruits (14%, 7/50). The most common drug triggers were antibiotics (59.4%, 22/37), non-steroidal anti-inflammatory drugs (NSAIDs) (10.8%, 4/37), vaccines (10.8%, 4/37), and herbal medicines (10.8%, 4/37). Common food allergens vary by age: milk and eggs for infants, fruit for children aged 1-2 years, and buckwheat for children older than 3 years. Ninety-eight cases (89.1%) had skin mucosal involvement, 78 (70.9%) had respiratory compromise, 45 (40.9%) had cardiovascular compromise, and 31 (28.2%) had gastrointestinal symptoms. Cardiovascular compromise and reduced level of consciousness were statistically different between the age groups (p < 0.05). Twenty cases (18.2%) had resolved spontaneously when they reached the hospital, 53 cases (48.1%) received epinephrine, 18 (16.4%) were hospitalized, and 2 (1.8%) experienced a biphasic reaction. CONCLUSIONS: Males are overrepresented in children with anaphylaxis, and food and drugs are common triggers in children. However, the types of common food triggers differ across age groups, and infants are more likely to have cardiovascular compromise and a reduced level of consciousness. However, the use of epinephrine remains inadequate. The training of clinical staff in recognizing anaphylaxis needs to be strengthened.
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Anafilaxia , Hipersensibilidade Alimentar , Lactente , Humanos , Criança , Masculino , Pré-Escolar , Adolescente , Feminino , Anafilaxia/etiologia , Estudos Retrospectivos , Hipersensibilidade Alimentar/diagnóstico , Epinefrina , Alérgenos , ChinaRESUMO
BACKGROUND: Airway remodeling, which contributes to the clinical course of childhood asthma, occurs due to airway inflammation and is featured by anomalous biological behaviors of airway smooth muscle cells (ASMCs). microRNA (miRNA) plays an essential role in the etiopathogenesis of asthma. OBJECTIVE: This research was aimed to characterize miR-506 in asthma and uncover potential regulatory machinery. MATERIAL AND METHODS: The asthmatic cell model was established by treating ASMCs with transforming growth factor-beta1 (TGF-ß1) and assessed by the levels of interleukin (IL)-1ß and interferon gamma (IFN-γ). Using real-time quantitative polymerase chain reaction, mRNA expression of miR-506 and polypyrimidine tract-binding protein 1 (PTBP1) was measured. Cell counting kit-8 and Transwell migration tests were used for estimating the capacity of ASMCs to proliferate and migrate. Luciferase reporter assay was used to corroborate whether miR-506 was directly bound to PTBP1. Expression of PTBP1, collagen I and III, and essential proteins of the wingless-related integration (Wnt)/ß-catenin pathway (ß-catenin, c-MYC and cyclin D1) was accomplished by Western blot analysis. The involvement of Wnt/ß-catenin signaling in asthma was confirmed by Wnt signaling pathway inhibitor (IWR-1). RESULTS: miR-506 was poorly expressed in asthmatic tissues and cell model. Functionally, overexpression of miR-506 reduced aberrant proliferation, migration, inflammation and collagen deposition of ASMCs triggered by TGF-ß1. Mechanically, miR-506 directly targeted the 3' untranslated region (3-UTR) of PTBP1 and had a negative regulation on PTBP1 expression. Moreover, overexpression of miR-506 suppressed the induction of Wnt/ß-catenin pathway. The administration of IWR-1 further validated negative correlation between miR-506 and the Wnt/ß-catenin pathway in asthma. CONCLUSION: Our data indicated that targeting miR-506/PTBP1/Wnt/ß-catenin axis might point in a helpful direction for treating asthma in children.
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Remodelação das Vias Aéreas , Asma , MicroRNAs , Criança , Humanos , Remodelação das Vias Aéreas/genética , Remodelação das Vias Aéreas/imunologia , Asma/genética , Asma/imunologia , Asma/patologia , beta Catenina/genética , beta Catenina/metabolismo , Proliferação de Células/genética , Ribonucleoproteínas Nucleares Heterogêneas/genética , Ribonucleoproteínas Nucleares Heterogêneas/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Proteína de Ligação a Regiões Ricas em Polipirimidinas/genética , Proteína de Ligação a Regiões Ricas em Polipirimidinas/metabolismo , Fator de Crescimento Transformador beta1/metabolismo , Via de Sinalização WntRESUMO
Asthma is a chronic airway disease involving airway inflammation and remodeling. Studies showed that tripartite motif-containing protein 33 (TRIM33) regulated natural immunity, inflammation, and pulmonary fibrosis. However, the role and regulatory mechanism of TRIM33 in children's asthma are unclear. In this study, the TRIM33 expressions in serum samples and platelet-derived growth factor BB (PDGF-BB)-induced airway smooth-muscle cells (ASMCs) were evaluated. A gain-of-function experiment was performed, and cell proliferation and migration were detected using CCK-8 and wound healing assays. Besides, the protein levels of EMT biomarkers and airway-remodeling markers were determined by Western blot assay. ELISA analyzed the contents of IL-1ß, IL-6, and TNF-α in the supernatant. The modulation of Smad4 expression and subsequent activation of Wnt/ß-catenin by TRIM33 were also assessed. We found that TRIM33 was downregulated in the serum from children who were asthma patients and PDGF-BB-induced ASMCs. TRIM33 overexpression showed decrease of PDGF-BB-induced ASMC proliferation and migration. Moreover, the augment of TRIM33 reduced the PDGF-BB-induced cell EMT and airway-remodeling marker levels and suppressed the secretions of inflammatory cytokines in PDGF-BB-induced ASMCs. Additionally, TRIM33 overexpression inhibited activation of Wnt/ß-catenin via reducing Smad4 expression to regulate asthma inflammation and airway remodeling. All in all, our study revealed that TRIM33 expression was downregulated in children who were asthma patients and PDGF-BB-induced ASMCs. TRIM33 modulated PDGF-BB-induced inflammation and airway remodeling of ASMCs by the Wnt/ß-catenin pathway via regulating Smad4, which may provide a new treatment direction for asthma.
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Remodelação das Vias Aéreas , Asma , Criança , Humanos , Remodelação das Vias Aéreas/fisiologia , Asma/metabolismo , Becaplermina/metabolismo , beta Catenina/metabolismo , Movimento Celular , Proliferação de Células , Células Cultivadas , Inflamação/metabolismo , Interleucina-6/metabolismo , Miócitos de Músculo Liso/metabolismo , Sincalida/metabolismo , Fatores de Transcrição , Fator de Necrose Tumoral alfa/metabolismo , Via de Sinalização WntRESUMO
OBJECTIVE: Thise study is aimed to identify the biomarkers for predicting refractory Mycoplasma pneumoniae pneumonia in Chinese children at the time of the hospital admission. METHODS: The case control study retrospectively analyzed the clinical characteristics and laboratory results of Chinese pediatric patients presenting with common and refractory Mycoplasma pneumoniae pneumonia (CMPP and RMPP). Overall, there were 216 cases in the CMPP group and 88 cases in the RMPP group. Venous blood was collected, and serum ferritin (SF), lactate dehydrogenase (LDH), D-dimer, C-reactive protein (CRP), procalcitonin (PCT), neutrophil count/lymphocyte count (NLR), and other indexes were measured. A single factor analysis, an ROC curve analysis, and a logistic regression analysis were used to determine the independent risk factors of RMPP and find combination of initial markers for RMPP. RESULTS: There were significant differences between the RMPP group and the CMPP group in mean SF (529.82 [357.86] vs. 147.22 [122.68] ng/mL), LDH (522.08 [389.08] vs. 286.85 [101.02] U/L), D-dimer (6.65 [5.66] vs. 1.46 [2.45] µg/mL), CRP (62.80 [52.15] vs. 19.03 [24.50] mg/L), PCT (0.80 [2.61] vs. 0.16 [0.44]) ng/mL, and NLR (4.14 [2.52] vs. 2.62 [1.55]), with P < 0.05 for each comparison. ROC cut-off values of the above indexes were 329.01 ng/mL, 375.50 U/L, 2.10 µg/mL, 43.08 mg/L, 0.08 ng/mL, and 2.96, respectively. The logistic regression analysis showed that SF, D-dimer, and CRP are independent risk factors to predict RMPP. CONCLUSION: SF, D-dimer, and CRP are statistically significant biomarkers to predict RMPP in Chinese children patients in the settings of pediatric emergency department.
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Antibacterianos/uso terapêutico , Farmacorresistência Bacteriana , Ferritinas/sangue , Produtos de Degradação da Fibrina e do Fibrinogênio/análise , Mycoplasma pneumoniae/efeitos dos fármacos , Pneumonia por Mycoplasma/diagnóstico , Pneumonia por Mycoplasma/tratamento farmacológico , Fatores Etários , Biomarcadores/sangue , Criança , Pré-Escolar , China , Feminino , Humanos , Masculino , Mycoplasma pneumoniae/patogenicidade , Admissão do Paciente , Pneumonia por Mycoplasma/sangue , Pneumonia por Mycoplasma/microbiologia , Valor Preditivo dos Testes , Receptores Imunológicos/análise , Estudos Retrospectivos , Medição de Risco , Fatores de Risco , Resultado do TratamentoRESUMO
Hypoxia-mediated neurotoxicity contributes to various neurodegenerative disorders, including Alzheimer's disease and multiple sclerosis. Tetramethylpyrazine (TMP), a major bioactive component purified from Ligusticum wallichii Franchat, exhibited potent neuroprotective effect. However, the mechanism of TMP-exerted neuroprotective effect against hypoxia was not clear. In the study, we investigated the mechanism of the neuroprotective effect of TMP against hypoxia induced by CoCl2 in vitro and in vivo. The results showed that TMP could protect against CoCl2 -induced neurotoxicity in PC12 cells and in rats, as evidenced by enhancement of cell viability in PC12 cells and improvement of learning and memory ability in rats treated with CoCl2 . TMP could inhibit mitochondrial dysfunction, mitochondrial apoptotic molecular events, and thus apoptosis induced by CoCl2 . TMP inhibited CoCl2 -increased reactive oxygen species (ROS) level, which may contribute to hypoxia-related neurotoxicity induced by CoCl2 . The antioxidant and neuroprotective activities of TMP involved two pathways: one was the enhancement of nuclear factor erythroid 2-related factor 2 (Nrf2)/catalytic subunit of γ-glutamylcysteine ligase-mediated regulation of GSH and the other was the inhibition of hypoxia-inducible factor 1 α/NADPH oxidase 2 (NOX2)-mediated ROS generation. These two pathways contributed to improvement of oxidative stress and thus the amelioration of apoptosis under hypoxic conditions. These results have appointed a new path toward the understanding of pathogenesis and TMP-related therapy of hypoxia-related neurodegenerative diseases. We proposed two cascades for tetramethylpyrazine-exhibited protective effects against CoCl2 -induced neurotoxicity: One is enhancement of nuclear factor erythroid 2-related factor 2-catalytic subunit of γ-glutamylcysteine ligase-mediated regulation of glutathone and the other was the inhibition of hypoxia-inducible factor 1 α-NADPH oxidase-2-mediated ROS generation. We think these findings should provide a new understanding of pathogenesis and tetramethylpyrazine-related therapy of hypoxia-related neurodegenerative diseases.
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Hipóxia Celular/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Pirazinas/farmacologia , Transdução de Sinais/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Western Blotting , Imunoprecipitação da Cromatina , Cobalto/toxicidade , Glutamato-Cisteína Ligase/metabolismo , Glutationa/metabolismo , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Masculino , Glicoproteínas de Membrana/metabolismo , NADPH Oxidase 2 , NADPH Oxidases/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Células PC12 , Ratos , Ratos Wistar , Espécies Reativas de Oxigênio/metabolismo , TransfecçãoRESUMO
Idiopathic membranous nephropathy (IMN), which was the result from non-SLE, non-hepatitis B or C virus infection, is relatively rare in children. We collected 584 pediatric cases of renal biopsy specimens in children hospitalized in Xijing Hospital from Jan 2008 to Dec 2013. We found that IgA nephropathy (27.9%) and Henoch-Schönlein purpura (26.4%) are the most common etiology of pediatric kidney disease, followed by minimal change disease (10.4%). Surprisingly, the prevalence of IMN is very high (7.5%) in pediatric patients. Moreover, the prevalence of IMN is quickly increasing from 4.4% in 2009 to 9.1% in 2013.
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Glomerulonefrite Membranosa/epidemiologia , Glomerulonefrite Membranosa/etiologia , Adolescente , Criança , China/epidemiologia , Feminino , Glomerulonefrite por IGA/complicações , Glomerulonefrite por IGA/epidemiologia , Humanos , Vasculite por IgA/complicações , Vasculite por IgA/epidemiologia , Masculino , Nefrose Lipoide/complicações , Nefrose Lipoide/epidemiologia , Prevalência , Estudos Retrospectivos , Adulto JovemRESUMO
BACKGROUND AND OBJECTIVE: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes acute necrotizing encephalopathy (ANE), which has a high mortality rate and severe sequelae. This study aimed to identify ANE early and evaluate the usefulness of tocilizumab in ANE treatment. METHODS: We retrospectively included eight paeefediatric ANE cases infected with SARS-CoV-2 at Xi'an Children's Hospital, China, from December 1, 2022 to May 1, 2023. A literature search was performed using the PUBMED, SPRING, SCOPUS, and EMBASE databases. This study included eleven patients. Clinical characteristics, laboratory test results, imaging features, and treatment options were analysed. RESULTS: Eight of the 19 cases (42 %) died, one (5 %) recovered, and nine (47 %) improved with residual neurological dysfunction. Eighteen patients presented with fever, with 56 % having ≥40 °C. Twelve patients (63 %) presented with dysfunction consciousness. Eight (42 %) patients experienced frequent convulsions. All eight patients in our hospital had elevated procalcitonin levels (mean: 21.32 ng/mL, range: 0.10-89.40 ng/mL). Alanine aminotransferase levels were elevated (mean: 632.81 U/L, range: 13.00-2251.00 U/L) in six patients. Seven patients showed elevated uric acid levels(mean: 396.50 µmol/L, range: 157.00-660.00 µmol/L). Brain imaging indicated that all the patients had symmetrical injuries to the bilateral thalami, accompanied by symmetrical injuries in the cerebrum, cerebellum, basal ganglia, and brain stem. Compared with the classical treatment (n = 9), the combination with tocilizumab (n = 6) showed a statistically difference in mortality (p = 0.028 < 0.05). CONCLUSION: The typical clinical manifestations of ANE in children with SARS-CoV-2 infection are acute onset with high fever, frequent convulsions and rapidly worsening disturbance of consciousness. Tocilizumab treatment could reduces mortality in ANE.
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INTRODUCTION: Incontinentia pigmenti (IP) is a rare X-linked dominant genetic disease affecting ectodermal tissue and often misdiagnosed in the neonatal period. The aim of this study was to highlight sequential clinical features and evaluate prognosis of the 32 neonatal IP patients. MATERIAL AND METHODS: A retrospective descriptive analysis was performed, using the clinical, blood analytical, pathological, radiological, genetic, and followed-up data of neonatal patients diagnosed with IP from 2010 to 2021, in Xi'an, China. RESULTS: Of the 32 patients, two (6.25%) were male. Thirty babies (93.75%) had eosinophilia (eosinophilic granulocyte count: 0.31-19.9 × 109 , mean proportion of white blood cells: 20.98 ± 15.21%). Twenty babies (62.5%) had thrombocytosis (thrombocyte count: 139-975 × 109 , mean count: 416.76 ± 176.82). Thirty-one babies (96.88%) exhibited the first three cutaneous lesion stages characterized by erythema and superficial vesicles on inflammatory bases in a linear distribution in the first week of age. Thirteen babies (40%) combined nervous system abnormalities, and nine babies (28.13%) had retinopathy. Two types of genetic mutations were detected in the NEMO gene. Nineteen babies were followed up. According to the follow-up, four babies displayed psychomotor retardation, and five babies developed a decrease in vision with astigmatism and amblyopia. CONCLUSION: It is important that 30 babies (93.75%) had eosinophilia and 20 babies (62.5%) had thrombocytosis. Therefore, we speculate that the mechanism of the injury may be related to the platelet aggregation on the basis of the increase in eosinophil cells and the release of inflammatory factors.
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Eosinofilia , Doenças Genéticas Ligadas ao Cromossomo X , Incontinência Pigmentar , Trombocitose , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , População do Leste Asiático , Incontinência Pigmentar/complicações , Incontinência Pigmentar/diagnóstico , Incontinência Pigmentar/genética , Estudos RetrospectivosRESUMO
BACKGROUND: Trauma is the leading cause of death among children worldwide. The inflammatory response of paediatric patients to multiple injuries can be monitored using serum interleukin-6 (IL-6) levels. This study aimed to assess the value of IL-6 levels in predicting the severity of paediatric trauma and its clinical association with disease activity. METHOD: We prospectively tested serum IL-6 levels and evaluated the Paediatric Trauma Score (PTS) and other clinical data among 106 paediatrics trauma patients from January 2022 to May 2023 at the Emergency Department of the Xi'an Children's Hospital in China. The relationship between IL-6 and trauma severity levels by PTS was analyzed statistically. RESULTS: IL-6 levels were elevated in 76 (71.70%) of the 106 paediatric patients with trauma. Spearman's test showed a significant negative linear correlation between IL-6 and PTS (rs = - 0.757, p < 0.001). IL-6 levels were moderate positively correlated with alanine aminotransferase, aspartate aminotransferase, white blood cells, blood lactic acid and interleukin 10 (rs = 0.513, 0.600, 0.503, 0.417, 0.558, p < 0.01). IL-6 levels were positively correlated with hypersensitive C-reactive protein and glucose (rs = 0.377, rs = 0.389, respectively, p < 0.001). IL-6 levels were negatively correlated with fibrinogen and PH (rs = - 0.434, p < 0.001; rs = - 0.382, respectively, p < 0.001). Binary scatter plots further demonstrated higher levels of IL-6 correlated with lower PTS scores. CONCLUSION: Serum IL-6 levels significantly increased with increasing severity of paediatric trauma. Serum levels of IL-6 can function as important indicators for predicting disease severity and activity in paediatric trauma patients.
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Hospitalização , Interleucina-6 , Criança , Humanos , Proteína C-Reativa , China , Gravidade do PacienteRESUMO
Soil salt-alkalization seriously impacts crop growth and productivity worldwide. Breeding and applying tolerant varieties is the most economical and effective way to address soil alkalization. However, genetic resources for breeders to improve alkali tolerance are limited in mung bean. Here, a genome-wide association study (GWAS) was performed to detect alkali-tolerant genetic loci and candidate genes in 277 mung bean accessions during germination. Using the relative values of two germination traits, 19 QTLs containing 32 SNPs significantly associated with alkali tolerance on nine chromosomes were identified, and they explained 3.6 to 14.6% of the phenotypic variance. Moreover, 691 candidate genes were mined within the LD intervals containing significant trait-associated SNPs. Transcriptome sequencing of alkali-tolerant accession 132-346 under alkali and control conditions after 24 h of treatment was conducted, and 2565 DEGs were identified. An integrated analysis of the GWAS and DEGs revealed six hub genes involved in alkali tolerance responses. Moreover, the expression of hub genes was further validated by qRT-PCR. These findings improve our understanding of the molecular mechanism of alkali stress tolerance and provide potential resources (SNPs and genes) for the genetic improvement of alkali tolerance in mung bean.
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Estudo de Associação Genômica Ampla , Vigna , Vigna/genética , Polimorfismo de Nucleotídeo Único , Melhoramento Vegetal , Locos de Características Quantitativas , RNA-Seq , SoloRESUMO
OBJECTIVES: To explore the impacts of cordycepin and underlying mechanism on the sepsis. METHODS: The sepsis mice model was built and treated with different concentrations of cordycepin. Then the liver and lung injury caused by cecal ligation and puncture (CLP) was assessed using H&E staining and TUNEL assay. The expression of relevant genes was detected using qRT-PCR analysis and ELISA assays. Besides, the macrophage polarization was checked by flow cytometry. KEY FINDINGS: Cordycepin could significantly improve the liver and lung injury. Moreover, cordycepin increased the distribution of F4/80+ CD206+ M2-like macrophages and F4/80+ iNOS+ M1-like macrophages through down-regulating the expression of relevant genes. More importantly, cordycepin could monitor the protein expression of iNOS, Arg-1, TNF-α, MCP-1, IL-4 and IL-10 in CLP mice. Meanwhile, the elevated level of p65 induced by CLP was also repressed by the increase of the cordycepin. Moreover, cordycepin played a crucial part in CLP mice through modulating the NF-κB/p65 signalling pathway. CONCLUSIONS: Cordycepin played an important role in mice with sepsis via reducing the M1/M2 macrophage polarization and modulating the NF-κB/p65 signalling pathway.
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Desoxiadenosinas/farmacologia , Macrófagos/efeitos dos fármacos , Sepse/tratamento farmacológico , Animais , Modelos Animais de Doenças , Hepatopatias/tratamento farmacológico , Hepatopatias/etiologia , Pneumopatias/tratamento farmacológico , Pneumopatias/etiologia , Macrófagos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , NF-kappa B/metabolismo , Sepse/complicações , Transdução de Sinais/efeitos dos fármacos , Fator de Transcrição RelA/metabolismoRESUMO
BACKGROUND: Circular RNAs (circRNAs) are a new type of stable noncoding RNA and have been proven to play a crucial role in osteoporosis. This study explored the role and mechanism of hsa_circ_0001485 in osteogenic differentiation. METHODS: Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis and Gene Ontology (GO) enrichment analysis were performed according to the previous sequencing data in human bone marrow mesenchymal stem cells (BMSC) before and after the induction of osteogenic differentiation on the differentially expressed circRNAs, to screen out signaling pathways associated with osteogenic differentiation. The hFOB 1.19 cells were used to verify the function and mechanism of specific circRNAs in osteogenic differentiation. Additionally, small interfering fragments and overexpression plasmids were used to determine the role of specific circRNAs during osteogenic differentiation. Furthermore, pull-down experiments and mass spectrometry were performed to determine the proteins that bind to specific circRNAs. RESULTS: The KEGG and GO enrichment analyses showed that the TGFß-BMP signaling pathway was related to the osteogenic differentiation process, and four circRNAs were associated with the pathway. The quantitative polymerase chain reaction analysis revealed that hsa_circ_0001485 expression was increased during the osteogenic differentiation process of BMSCs. Knockdown of hsa_circ_0001485 suppressed the activity of the alkaline phosphatase enzyme and the expression of RUNX2, osteopontin, and osteocalcin in the osteogenic hFOB 1.19 cells, whereas overexpression of hsa_circ_0001485 promoted their expression. Additionally, we found that hsa_circ_0001485 and BMPR2 targeted binding to activate the TGFß-BMP signaling pathway and promoted osteogenic differentiation through mass spectrometry analysis. CONCLUSION: This study demonstrates that hsa_circ_0001485 is highly expressed in the osteogenic hFOB 1.19 cells, which activate the TGFß-BMP pathway through targeted binding of BMPR2, and plays a positive role in regulating osteogenic differentiation.
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Receptores de Proteínas Morfogenéticas Ósseas Tipo II , Células-Tronco Mesenquimais , MicroRNAs , Osteogênese , RNA Circular , Fator de Crescimento Transformador beta , Receptores de Proteínas Morfogenéticas Ósseas Tipo II/genética , Receptores de Proteínas Morfogenéticas Ósseas Tipo II/metabolismo , Diferenciação Celular/genética , Diferenciação Celular/fisiologia , Humanos , Células-Tronco Mesenquimais/metabolismo , MicroRNAs/metabolismo , Osteogênese/genética , Osteogênese/fisiologia , RNA Circular/genética , RNA Circular/metabolismo , Transdução de Sinais/genética , Transdução de Sinais/fisiologia , Fator de Crescimento Transformador beta/genética , Fator de Crescimento Transformador beta/metabolismoRESUMO
The cleaning potential of selected Chinese bituminous coals during coal preparation was evaluated in terms of coal quality, cleaning grade, and cleanability. The cleaning potentials were determined for coals sampled from a total of six seams, two each in three different general exploration areas in southern Shanxi, China. Distribution maps of resources with different cleaning potential characteristics were prepared using mapping and geographical information software. In each case, the area and calculated reserves for coals having different classifications of cleaning potentials were determined. The total areal extent of the six coals studied is about 410 million m2, and the total reserves of these coals amount to about 1460 million tonnes, which include coals of poor, fair, good, and proficient cleanabilities. The proportion of high-quality coals is about 28.9%. Coals that can be processed into high-quality coals account for about 69.2%. A cleaning potential gradient is proposed for indicating the cleaning potential level, and an equation for calculating it is established. The necessity of processing raw coal of good quality is also to be considered in terms of economics. It does not make economic sense to process coal with an organic sulfur content more than 1.5%, even though it may have a good cleanability.
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The present study aimed to assess the role of miR-1275 in cardiac ischemia reperfusion injury. H9 human embryonic stem cell (hESC)-derived cardiomyocytes stimulated by oxygen-glucose deprivation/reoxygenation (OGD/R) were used to simulate myocardial injury in vitro. miR-1275 expression levels in cells were measured by RT-qPCR. The release of lactate dehydrogenase (LDH) and creatine kinase (CK) was examined through LDH and CK ELISA kits. Cell apoptosis was detected through flow cytometry. A Fura-2 Calcium Flux Assay Kit and a Fluo-4 assay kit were used to determine the Ca2+ concentration. Expression levels of proteins were tested by Western blotting. The binding effect of miR-1275 and neuromedin U type 1 receptor (NMUR1) was detected by dual-luciferase activity assay. The results showed that miR-1275 was upregulated in OGD/R-stimulated cardiomyocytes. Inhibition of miR-1275 suppressed the increased activity of LDH and CK, cell apoptosis, reactive oxygen species (ROS) production, intracellular Ca2+ concentration and sarcoplasmic reticulum (SR) Ca2+ leak induced by OGD/R treatment in cardiomyocytes. miR-1275 directly targets 3'UTR of NMUR1 and negatively regulates NMUR1 expression. Silence of NMUR1 abolished the protecting effect of the miR-1275 antagomir on myocardial OGD/R injury. Our study indicated that the miR-1275 antagomir protects cardiomyocytes from OGD/R injury through the promotion of NMUR1.
Assuntos
Sinalização do Cálcio/genética , Técnicas de Silenciamento de Genes/métodos , MicroRNAs/genética , MicroRNAs/metabolismo , Traumatismo por Reperfusão Miocárdica/metabolismo , Miócitos Cardíacos/metabolismo , Receptores de Neurotransmissores/metabolismo , Regiões 3' não Traduzidas/genética , Animais , Antagomirs/farmacologia , Apoptose/genética , Cálcio/metabolismo , Hipóxia Celular/genética , Linhagem Celular , Células Alimentadoras/metabolismo , Fibroblastos/metabolismo , Glucose/metabolismo , Células-Tronco Embrionárias Humanas/citologia , Humanos , Camundongos , Traumatismo por Reperfusão Miocárdica/genética , Estresse Oxidativo/genética , Oxigênio/metabolismo , Substâncias Protetoras/farmacologia , Ratos , Espécies Reativas de Oxigênio/metabolismo , Receptores de Neurotransmissores/genética , Regulação para Cima/genéticaRESUMO
AIMS: Pediatric heart failure is a common cardiovascular disease in clinical pediatrics. CCCTC-binding factor (CTCF), a novel transcriptional repressor, was reported to participate in the occurrence of various cardiovascular diseases. The present study focuses on exploring the effects of CTCF on tunicamycin (TM)-induced endoplasmic reticulum (ER) stress, and investigating the underlying mechanisms. MATERIALS AND METHOD: Expression of CTCF in blood samples of heart failure children and TM-induced cardiomyocytes were evaluated by real-time quantitative PCR (RT-qPCR). Apoptotic rate of cardiomyocytes was detected by Annexin v assay. Western blotting and enzyme-linked immunosorbent assay (ELISA) were applied to examine the effect of CTCF on ER stress. Co-immunoprecipitation and western blotting were devoted to explore the mechanism by which CTCF contributes to ER stress. KEY FINDINGS: We proved that CTCF was lowly expressed in blood samples of heart failure children and TM-induced cardiomyocytes, and overexpression of CTCF weaken the TM-induced ER stress. Using co-immunoprecipitation and protein blots, we demonstrated that CTCF upregulates RYR2 by inhibiting S100A1, thus mediating the PERK signaling pathway and regulating ER stress. SIGNIFICANCE: Our data revealed that CTCF protects cardiomyocytes from ER stress through S100A1-RYR2 axis, and can be applied as a therapeutic target for the treatment of pediatric heart failure in future.
Assuntos
Apoptose/fisiologia , Fator de Ligação a CCCTC/fisiologia , Estresse do Retículo Endoplasmático/fisiologia , Miócitos Cardíacos/efeitos dos fármacos , Canal de Liberação de Cálcio do Receptor de Rianodina/metabolismo , Proteínas S100/metabolismo , Adolescente , Animais , Western Blotting , Fator de Ligação a CCCTC/metabolismo , Criança , Pré-Escolar , Ensaio de Imunoadsorção Enzimática , Feminino , Insuficiência Cardíaca/metabolismo , Insuficiência Cardíaca/fisiopatologia , Humanos , Imunoprecipitação , Masculino , Camundongos , Miócitos Cardíacos/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Canal de Liberação de Cálcio do Receptor de Rianodina/fisiologia , Proteínas S100/fisiologia , Regulação para CimaRESUMO
Apoptosis of cardiomyocytes and oxidant stress are considered essential processes in the progression of cardiovascular diseases. A hypoxic stress which causes apoptosis of cardiomyocytes is the main problem in ischemic heart disease. The aim of the present study was to explore the functional role and potential mechanisms of miR-223-3p in hypoxia-induced cardiomyocyte apoptosis and oxidative stress. Here, we observed a increment of miR-223-3p level accompanied by the decrease of Krüppel-like zinc-finger transcription factor 15 (KLF15) expression in response to hypoxia. Additionally, absence of miR-223-3p manifestly dampened hypoxia-induced cardiomyocyte injury in H9c2 cells, including improving cell viability, attenuating the LDH leakage and preventing cardiomyocyte apoptosis accompanied by an increase in the expression of Bcl-2 and a decrease in the expression of Bax and C-caspase 3 in the setting of hypoxia. Moreover, depletion of miR-223-3p evidently retarded oxidant stress by inhibiting reactive oxygen species generation and lipid peroxidation, as well as enhancing antioxidant enzyme activity in H9c2 cells following exposure to hypoxia. More importantly, KLF15 was a direct and functional target of miR-223-3p. Further data validated that miR-223-3p negatively regulated the expression of KLF15. Mechanistically, deletion of KLF15 partly abrogated the suppressive effects of miR-223-3p deletion on hypoxia-induced cardiomyocyte apoptosis and oxidative stress. Taken all data together, our findings established that our study defines a novel mechanism by which miR-223-3p protects against cardiomyocyte apoptosis and oxidative stress by targeting KLF15, suggesting that the miR-223-3p/KLF15 may be a potential therapeutic target for ischemic heart conditions.