RESUMO
Intestinal stem cells (ISCs) at the crypt base are responsible for the regeneration of the intestinal epithelium. However, how ISC self-renewal is regulated still remains unclear. Here we identified a circular RNA, circBtnl1, that is highly expressed in ISCs. Loss of circBtnl1 in mice enhanced ISC self-renewal capacity and epithelial regeneration, without changes in mRNA and protein levels of its parental gene Btnl1. Mechanistically, circBtnl1 and Atf4 mRNA competitively bound the ATP-dependent RNA helicase Ddx3y to impair the stability of Atf4 mRNA in wild-type ISCs. Furthermore, ATF4 activated Sox9 transcription by binding to its promoter via a unique motif, to enhance the self-renewal capacity and epithelial regeneration of ISCs. In contrast, circBtnl1 knockout promoted Atf4 mRNA stability and enhanced ATF4 expression, which caused Sox9 transcription to potentiate ISC stemness. These data indicate that circBtnl1-mediated Atf4 mRNA decay suppresses Sox9 transcription that negatively modulates self-renewal maintenance of ISCs.
Assuntos
Fator 4 Ativador da Transcrição , Mucosa Intestinal , Estabilidade de RNA , RNA Circular , RNA Mensageiro , Regeneração , Células-Tronco , Células-Tronco/citologia , Células-Tronco/fisiologia , Organoides/citologia , Camundongos Endogâmicos C57BL , Animais , Camundongos , RNA Circular/genética , RNA Circular/metabolismo , Mucosa Intestinal/citologia , Mucosa Intestinal/fisiologia , Regeneração/genética , Fator 4 Ativador da Transcrição/genética , Fator 4 Ativador da Transcrição/metabolismo , RNA Mensageiro/metabolismo , Ativação Transcricional , Fatores de Transcrição SOX9/genética , Antígenos de Histocompatibilidade Menor/metabolismo , RNA Helicases DEAD-box/metabolismoRESUMO
Single nucleotide variants in the general population are common genomic alterations, where the majority are presumed to be silent polymorphisms without known clinical significance. Using human induced pluripotent stem cell (hiPSC) cerebral organoid modeling of the 1.4 megabase Neurofibromatosis type 1 (NF1) deletion syndrome, we previously discovered that the cytokine receptor-like factor-3 (CRLF3) gene, which is co-deleted with the NF1 gene, functions as a major regulator of neuronal maturation. Moreover, children with NF1 and the CRLF3L389P variant have greater autism burden, suggesting that this gene might be important for neurologic function. To explore the functional consequences of this variant, we generated CRLF3L389P-mutant hiPSC lines and Crlf3L389P-mutant genetically engineered mice. While this variant does not impair protein expression, brain structure, or mouse behavior, CRLF3L389P-mutant human cerebral organoids and mouse brains exhibit impaired neuronal maturation and dendrite formation. In addition, Crlf3L389P-mutant mouse neurons have reduced dendrite lengths and branching, without any axonal deficits. Moreover, Crlf3L389P-mutant mouse hippocampal neurons have decreased firing rates and synaptic current amplitudes relative to wild type controls. Taken together, these findings establish the CRLF3L389P variant as functionally deleterious and suggest that it may be a neurodevelopmental disease modifier.
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Células-Tronco Pluripotentes Induzidas , Criança , Humanos , Animais , Camundongos , Células-Tronco Pluripotentes Induzidas/metabolismo , Neurônios/metabolismo , Encéfalo/metabolismo , Receptores de Citocinas/metabolismo , Nucleotídeos/metabolismoRESUMO
Morphine and morphine-6-glucuronide (M6G) produce central nervous system (CNS) effects by activating mu-opioid receptors, while naloxone is used mainly for the reversal of opioid overdose, specifically for the fatal complication of respiratory depression, but also for alleviating opioid-induced side effects. In this study we developed a physiologically-based pharmacokinetic-pharmacodynamic (PBPK-PD) model to simultaneously predict pharmacokinetics and CNS effects (miosis, respiratory depression and analgesia) of morphine as well as antagonistic effects of naloxone against morphine. The pharmacokinetic and pharmacodynamic parameters were obtained from in vitro data, in silico, or animals. Pharmacokinetic and pharmacodynamic simulations were conducted using 39 and 36 clinical reports, respectively. The pharmacokinetics of morphine and M6G following oral or intravenous administration were simulated, and the PBPK-PD model was validated using clinical observations. The Emax model correlated CNS effects with free concentrations of morphine and M6G in brain parenchyma. The predicted CNS effects were compared with observations. Most clinical observations fell within the 5th-95th percentiles of simulations based on 1000 virtual individuals. Most of the simulated area under the concentration-time curve or peak concentrations also fell within 0.5-2-fold of observations. The contribution of morphine to CNS effects following intravenous or oral administration was larger than that of M6G. Pharmacokinetics and antagonistic effects of naloxone on CNS effects were also successfully predicted using the developed PBPK-PD model. In conclusion, the pharmacokinetics and pharmacodynamics of morphine and M6G, antagonistic effects of naloxone against morphine-induced CNS effects may be successfully predicted using the developed PBPK-PD model based on the parameters derived from in vitro, in silico, or animal studies.
Assuntos
Modelos Biológicos , Morfina , Naloxona , Antagonistas de Entorpecentes , Naloxona/farmacocinética , Naloxona/farmacologia , Humanos , Morfina/farmacocinética , Morfina/administração & dosagem , Morfina/farmacologia , Antagonistas de Entorpecentes/farmacocinética , Antagonistas de Entorpecentes/farmacologia , Antagonistas de Entorpecentes/administração & dosagem , Animais , Derivados da Morfina/farmacocinética , Sistema Nervoso Central/efeitos dos fármacos , Sistema Nervoso Central/metabolismo , Analgésicos Opioides/farmacocinética , Analgésicos Opioides/administração & dosagem , Masculino , Simulação por Computador , Administração Oral , Adulto , Administração Intravenosa , FemininoRESUMO
Conventional electrochemical activation of peroxymonosulfate (PMS) is not very cost-effective and practical by the excessive input of energy. The electricity generated by photosynthetic microalgae fuel cells (MFCs) is utilized to activate PMS, which would achieve the combination of green bioelectricity and advanced oxidation processes for sustainable pollutants degradation. In this study, a novel dual-chamber of MFCs was constructed by using microalgae as anode electron donor and PMS as cathode electron acceptor, which was operating under both close-circuit and open-circuit conditions. Under close-circuit condition, 1-12 mM PMS in cathode was successfully in situ activated, where 32.00%-99.83% of SMX was removed within 24 h, which was about 1.21-1.78 times of that in the open-circuit of MFCs. Meanwhile, a significant increase in bioelectricity generation in MFCs was observed after the accumulation of microalgae biomass (4.65-5.37 mg/L), which was attributed to the efficient electron separation and transfer. Furthermore, the electrochemical analysis demonstrated that SMX or its products were functioned as electronic shuttles, facilitating the electrochemical reaction and altering the electrical capacitance. The quenching experiments and voltage output results reflected that complex active radical (SO4â -, â OH, and 1O2) were involved in SMX removal. Seven degradation products of SMX were detected and S-N bond cleavage was the main degradation pathway. Predicted toxicity values calculated by ECOSAR program showed that all the products were less toxic or nontoxic. Finally, the density functional theory (DFT) calculations revealed that the O and N atoms on SMX were more susceptible to electrophilic reactions, which were more vulnerable to be attacked by reactive species. This study provided new insights into the activation of PMS by bioelectricity for SMX degradation, proposing the mechanisms for PMS activation and degradation sites of SMX.
Assuntos
Fontes de Energia Bioelétrica , Sulfametoxazol , Poluentes Químicos da Água , Poluentes Químicos da Água/química , Sulfametoxazol/química , Peróxidos/química , Microalgas/efeitos dos fármacos , Microalgas/química , OxirreduçãoRESUMO
BACKGROUND: It is imperative to use a validated tool to measure and understand health behavior as it greatly impacts health status. It has been demonstrated that the Health Behavior Inventory - Short Form (HBI-SF) is valid in several countries. The purpose of this study was to translate the HBI-SF into Chinese and test its psychometric properties. METHODS: This study employed a two-phase methodology. The phase one entailed the cross-cultural adaptation of the HBI-SF, while the subsequent phase evaluated the psychometric properties of the scale. This evaluation encompassed classical test theory (CTT) and item response theory (IRT) tests to assess reliability and validity. It involved 1058 participants, of whom 1036 completed the questionnaire successfully. Out of these, 518 were analyzed for CTT, along with 518 for IRT. RESULTS: The S-CVI was found to be 0.935, while the I-CVI ranged from 0.889 to 1.000. The results of the confirmatory factor analysis suggested the goodness-of-fit indices for the four-factor model was acceptable. Regarding the subscales, the average variance extracted (AVE) and Heterotrait-Monotrait Ratio of Correlations (HTMT) matrix also met the cutoff values (AVE > 0.5 and HTMI < 0.85). Furthermore, the internal consistency and composite reliability indices of all factors were greater than 0.7. Infit and/or outfit values indicated that all items fitted the Rasch model. The Wright map revealed that the average person measures for the participants (mean = - 0.994, SD = 0.430) were comparatively lower than the average item measures (mean = 0.000, SD = 0.257). The person separation reliability values for the four factors ranged from 0.600 to 0.746, indicating an acceptable level of reliability. Two items showed differential item functioning. CONCLUSIONS: The findings derived from both CTT and IRT analyses demonstrate favorable levels of reliability and validity for the Chinese version of the HBI-SF.
Assuntos
Comportamentos Relacionados com a Saúde , Psicometria , Humanos , Feminino , Masculino , Adulto , China , Reprodutibilidade dos Testes , Pessoa de Meia-Idade , Inquéritos e Questionários/normas , Análise Fatorial , Adulto Jovem , Traduções , AdolescenteRESUMO
Receptor-mediated vesicular transport has been extensively developed to penetrate the blood-brain barrier (BBB) and has emerged as a class of powerful brain-targeting delivery technologies. However, commonly used BBB receptors such as transferrin receptor and low-density lipoprotein receptor-related protein 1, are also expressed in normal brain parenchymal cells and can cause drug distribution in normal brain tissues and subsequent neuroinflammation and cognitive impairment. Here, the endoplasmic reticulum residing protein GRP94 is found upregulated and relocated to the cell membrane of both BBB endothelial cells and brain metastatic breast cancer cells (BMBCCs) by preclinical and clinical investigations. Inspired by that Escherichia coli penetrates the BBB via the binding of its outer membrane proteins with GRP94, avirulent DH5α outer membrane protein-coated nanocapsules (Omp@NCs) are developed to cross the BBB, avert normal brain cells, and target BMBCCs via recognizing GRP94. Embelin (EMB)-loaded Omp@EMB specifically reduce neuroserpin in BMBCCs, which inhibits vascular cooption growth and induces apoptosis of BMBCCs by restoring plasmin. Omp@EMB plus anti-angiogenic therapy prolongs the survival of mice with brain metastases. This platform holds the translational potential to maximize therapeutic effects on GRP94-positive brain diseases.
Assuntos
Neoplasias Encefálicas , Nanocápsulas , Camundongos , Animais , Células Endoteliais/metabolismo , Biomimética , Encéfalo/metabolismo , Neoplasias Encefálicas/tratamento farmacológico , Neoplasias Encefálicas/metabolismo , Proteínas de Membrana/metabolismo , Barreira Hematoencefálica/metabolismoRESUMO
BACKGROUND: Geographic differences exist in the antibiotic resistance patterns of Helicobacter pylori. Personalized treatment regimens based on local or individual resistance data are essential. We evaluated the current status of H. pylori resistance in Ningxia, analyzed resistance-related factors, and assessed the concordance of phenotypic and genotypic resistance. METHODS: Strains were isolated from the gastric mucosa of patients infected with H. pylori in Ningxia and relevant clinical information was collected. Phenotypic antibiotic susceptibility assays (Kirby-Bauer disk diffusion) and antibiotic resistance gene detection (Sanger sequencing) were performed. RESULTS: We isolated 1955 H. pylori strains. The resistance rates of H. pylori to amoxicillin, levofloxacin, clarithromycin, and metronidazole were 0.9%, 42.4%, 40.4%, and 94.2%, respectively. Only five tetracycline-resistant and one furazolidone-resistant strain were identified. Overall, 3.3% of the strains were sensitive to all six antibiotics. Multidrug-resistant strains accounted for 22.9%, of which less than 20% were from Wuzhong. Strains isolated from women and patients with nonulcerative disease had higher rates of resistance to levofloxacin and clarithromycin. Higher rates of resistance to metronidazole, levofloxacin, and clarithromycin were observed in the older age group than in the younger age group. The kappa coefficients of phenotypic resistance and genotypic resistance for levofloxacin and clarithromycin were 0.830 and 0.809, respectively, whereas the remaining antibiotics showed poor agreement. CONCLUSION: H. pylori antibiotic resistance is severe in Ningxia. Therefore, furazolidone, amoxicillin, and tetracycline are better choices for the empirical therapy of H. pylori infection in this region. Host sex, age, and the presence of ulcerative diseases may affect antibiotic resistance of the bacteria. Personalized therapy based on genetic testing for levofloxacin and clarithromycin resistance may be a future direction for the eradication therapy of H. pylori infection in Ningxia.
Assuntos
Infecções por Helicobacter , Helicobacter pylori , Humanos , Feminino , Idoso , Claritromicina/farmacologia , Claritromicina/uso terapêutico , Metronidazol/farmacologia , Metronidazol/uso terapêutico , Levofloxacino/farmacologia , Levofloxacino/uso terapêutico , Infecções por Helicobacter/tratamento farmacológico , Infecções por Helicobacter/microbiologia , Estudos Retrospectivos , Furazolidona/uso terapêutico , Testes de Sensibilidade Microbiana , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Amoxicilina/uso terapêutico , Tetraciclina/farmacologia , Tetraciclina/uso terapêutico , Resistência Microbiana a Medicamentos , Farmacorresistência BacterianaRESUMO
Virus infection is the key constraint to potato cultivation worldwide. Especially, coinfection by multiple viruses could exacerbate the yield loss. Transgenic plants expressing artificial microRNAs (amiRNAs) have been shown to confer specific resistance to viruses. In this study, three amiRNAs containing Arabidopsis miR159 as a backbone, expressing genes targeting P25, HC-Pro and Brp1 of potato virus X (PVX), potato virus Y (PVY) and potato spindle tuber viroid (PSTVd), were constructed. amiR-159P25, amiR-159HCPro and amiR-159Brp1 were cloned into the plant expression vector pCAMBIA1301 with a CaMV35S promoter, producing the p1301-pre-amiRP25-HCPro-Brp1 vector. Twenty-three transgenic plants (Solanum tuberosum cv. 'Youjin') were obtained by Agrobacterium tumefaciens-mediated transformation, and ten PCR-positive transplants were chosen for further analysis. Quantitative real-time PCR results indicated that 10 transgenic plants could express amiRNAs successfully. Southern blotting hybridization proved that amiR-159P25-HCPro-Brp1 had integrated into potato genome in transgenic lines. Viral (viroid) challenge assays revealed that these transgenic plants demonstrated resistance against PVX, PVY and PSTVd coinfection simultaneously, whereas the untransformed controls developed severe symptoms. This study demonstrates a novel amiRNA-based mechanism that may have the potential to develop multiple viral resistance strategies in potato.
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INTRODUCTION: Electroacupuncture (EA) treatment has been demonstrated to have the potential to prevent sepsis-induced hippocampal injury; however, the mechanisms underlying the protective effects of EA against such injury remain unclear. Herein, to elucidate these mechanisms, we constructed a mouse model of lipopolysaccharide (LPS)-induced hippocampal injury to investigate the protection mechanism of EA and to determine whether heme oxygenase-1 (HO-1)-mediated mitochondrial function is involved in the protective effect of EA. MATERIALS AND METHODS: The sepsis model of hippocampal injury was induced by administering LPS. The Zusanli and Baihui acupoints were stimulated using EA for 30 min once a day, for 5 d before LPS exposure and the first day after administering LPS. Hippocampal injury was investigated by hematoxylin and eosin staining and Nissl staining. HO-1 levels were measured using Western blotting. Mitochondrial metabolism was validated by assessing adenosine triphosphate, superoxide dismutase, malondialdehyde levels, reactive oxygen species production, and mitochondrial respiratory chain activity. Mitochondrial morphology was analyzed by transmission electron microscopy. RESULTS: EA treatment alleviated neuronal injury, impeded oxidative stress, and improved mitochondrial respiratory function, energy metabolism, and mitochondrial morphology in LPS-exposed mice. In addition, HO-1 knockout aggravated LPS-induced hippocampal injury, aggravated oxidative stress, and reduced mitochondrial respiratory function and aggravated mitochondrial swelling, crest relaxation, and vacuole degeneration. Moreover, EA was unable to reverse the hippocampal damage and mitochondrial dysfunction caused by LPS exposure after HO-1 knockout. CONCLUSIONS: EA improves LPS-induced hippocampal injury by regulating HO-1-mediated mitochondrial function. Furthermore, HO-1 plays a critical role in maintaining mitochondrial function and resisting oxidative injury.
Assuntos
Eletroacupuntura , Sepse , Animais , Heme Oxigenase-1/metabolismo , Hipocampo/metabolismo , Lipopolissacarídeos , Camundongos , Mitocôndrias/metabolismo , Estresse Oxidativo , Sepse/metabolismo , Sepse/terapiaRESUMO
Tissue regeneration is the preferred treatment for dentin and bone tissue defects. Dental pulp stem cells (DPSCs) have been extensively studied for their use in tissue regeneration, including the regeneration of dentin and bone tissue. LIM mineralization protein-1 (LMP-1) is an intracellular non-secretory protein that plays a positive regulatory role in the mineralization process. In this study, an LMP-1-induced DPSCs model was used to explore the effect of LMP-1 on the proliferation and odonto/osteogenic differentiation of DPSCs, as well as the underlying mechanisms. As indicated by the cell counting kit-8 assay, the results showed that LMP-1 did not affect the proliferation of DPSCs. Overexpression of LMP-1 significantly promoted the committed differentiation of DPSCs and vice versa, as shown by alkaline phosphatase activity assay, alizarin red staining, western blot assay, quantitative real-time polymerase chain reaction assay, and in vivo mineralized tissue formation assay. Furthermore, inhibiting the activation of the extracellular signal-regulated kinase 1/2 (ERK1/2), p38 mitogen-activated protein kinase (MAPK), and c-Jun N-terminal kinase (JNK) pathways using specific pathway inhibitors showed that the ERK1/2 and p38 MAPK pathways attenuated the differentiation of DPSCs. Besides, the expression of BMP signaling pathway components were also determined, which suggested that LMP-1 could activate BMP-2/Smad1/5 signaling pathway. Our results not only indicated the underlying mechanism of LMP-1 treated DPSCs but also provided valuable insight into therapeutic strategies in regenerative medicine.
Assuntos
Osteogênese , Proteínas Quinases p38 Ativadas por Mitógeno , Diferenciação Celular , Proliferação de Células/genética , Células Cultivadas , Polpa Dentária/metabolismo , Sistema de Sinalização das MAP Quinases , Proteína Quinase 3 Ativada por Mitógeno , Osteogênese/genética , Transdução de Sinais , Células-Tronco/fisiologia , Proteínas Quinases p38 Ativadas por Mitógeno/genética , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
LncRNAs are associated with tumorigenesis of liver cancer. LncRNA Colorectal Neoplasia Differentially Expressed (CRNDE) was identified as an oncogenic lncRNA and involved in tumor growth and metastasis. The role of CRNDE in liver cancer was investigated. CRNDE was elevated in liver cancer cells. Knockdown of CRNDE decreased cell viability and inhibited proliferation of liver cancer. Moreover, knockdown of CRNDE reduced levels of extracellular acidification rate, glucose consumption, and lactate production to repress glycolysis of liver cancer. Silence of CRNDE enhanced the expression of miR-142 and reduced enhancer of zeste homolog 2 (EZH2) and hypoxia-inducible factor 1α (HIF-1α). Over-expression of HIF-1α attenuated CRNDE silence-induced decrease of glucose consumption and lactate production. Injection with sh-CRNDE virus reduced in vivo tumor growth of liver cancer through up-regulation of miR-142 and down-regulation of EZH2 and HIF-1α. In conclusion, knockdown of CRNDE suppressed cell proliferation, glycolysis, and tumor growth of liver cancer through EZH2/miR-142/HIF-1α.
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Neoplasias Colorretais , Neoplasias Hepáticas , MicroRNAs , RNA Longo não Codificante , Humanos , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Linhagem Celular Tumoral , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patologia , Neoplasias Colorretais/genética , Neoplasias Colorretais/patologia , MicroRNAs/genética , MicroRNAs/metabolismo , Lactatos , Glicólise , Hipóxia/genética , Proliferação de Células , Regulação Neoplásica da Expressão GênicaRESUMO
The mitotic phase is a vital step in cell division and may be involved in cancer progression, but it remains unclear whether genetic variants in mitotic phase-related pathways genes impact the survival of these patients. Here, we investigated associations between 31 032 single nucleotide polymorphisms (SNPs) in 368 mitotic phase-related pathway genes and overall survival (OS) of patients with nonsmall cell lung cancer (NSCLC). We assessed the associations in a discovery data set of 1185 NSCLC patients from the Prostate, Lung, Colorectal and Ovarian (PLCO) Cancer Screening Trial and validated the findings in another data set of 984 patients from the Harvard Lung Cancer Susceptibility Study. As a result, we identified three independent SNPs (ie, CHEK1 rs76744140 T>C, PRIM2 rs6939623 G>T and CDK6 rs113181986 G>C) to be significantly associated with NSCLC OS with an adjusted hazard ratio of 1.29 (95% confidence interval = 1.11-1.49, P = 8.26 × 10-4 ), 1.26 (1.12-1.42, 1.10 × 10-4 ) and 0.73 (0.63-0.86, 1.63 × 10-4 ), respectively. Moreover, the number of combined unfavorable genotypes of these three SNPs was significantly associated with NSCLC OS and disease-specific survival in the PLCO data set (Ptrend < .0001 and .0003, respectively). Further expression quantitative trait loci analysis showed that the rs76744140C allele predicted CHEK1 mRNA expression levels in normal lung tissues and that rs113181986C allele predicted CDK6 mRNA expression levels in whole blood tissues. Additional analyses indicated CHEK1, PRIM2 and CDK6 may impact NSCLC survival. Taken together, these findings suggested that these genetic variants may be prognostic biomarkers of patients with NSCLC.
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Carcinoma Pulmonar de Células não Pequenas/mortalidade , Quinase 1 do Ponto de Checagem/genética , Quinase 6 Dependente de Ciclina/genética , DNA Primase/genética , Neoplasias Pulmonares/mortalidade , Polimorfismo de Nucleotídeo Único , Carcinoma Pulmonar de Células não Pequenas/genética , Bases de Dados Factuais , Feminino , Regulação Neoplásica da Expressão Gênica , Estudo de Associação Genômica Ampla , Humanos , Neoplasias Pulmonares/genética , Masculino , Prognóstico , Modelos de Riscos Proporcionais , Locos de Características Quantitativas , Análise de SobrevidaRESUMO
Schisantherin A is an active ingredient originating from Schisandra chinensis (Turcz.) which has hepatoprotective and anti-oxidation activities. In this study, in vitro metabolisms investigated on rat liver microsomes (RLMs) and in vivo metabolisms explored on male Sprague Dawley rats of Schisantherin A were tested, respectively. The metabolites of Schisantherin A were identified using ultra-high-performance liquid chromatography coupled with hybrid triple quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS/MS). Based on the method, 60 metabolites were successfully identified and structurally characterized including 48 phase-I and 12 phase-II metabolites. Among the metabolites, 45 metabolites were reported for the first time. Moreover, 56 and eight metabolites were detected in urine and bile and 19 metabolites were identified in rats' plasma. It demonstrated that hepatic and extra-hepatic metabolic pathways were both involved in Schisantherin A biotransformation in rats. Five in vitro metabolites were structurally characterized for the first time. The results indicated that the metabolic pathways mainly include oxidation, reduction, methylation, and conjugation with glucuronide, taurine, glucose, and glutathione groups. This study provides a practical strategy for rapidly screening and identifying metabolites, and the results provide basic data for future pharmacological and toxicology studies of Schisantherin A and other lignin ingredients.
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Ciclo-Octanos/análise , Ciclo-Octanos/metabolismo , Dioxóis/análise , Dioxóis/metabolismo , Avaliação Pré-Clínica de Medicamentos , Lignanas/análise , Lignanas/metabolismo , Metaboloma , Espectrometria de Massas em Tandem , Animais , Cromatografia Líquida de Alta Pressão , Ciclo-Octanos/química , Dioxóis/química , Íons , Lignanas/química , Masculino , Redes e Vias Metabólicas , Metabolômica , Oxirredução , Ratos Sprague-DawleyRESUMO
BACKGROUND: Long noncoding RNA (lncRNA) is emerging as a vital regulator in various cancers. Recently, it was found that lncRNA colorectal neoplasia differentially expressed (CRNDE) plays an oncogenic role, promoting cell proliferation and migration in hepatocellular carcinoma (HCC). However, the underlying regulatory mechanism of lncRNA CRNDE remains unclear. METHODS: The expression levels of lncRNA CRNDE and miR-337-3p were analyzed by real-time polymerase chain reaction, and sineoculis homeobox homolog 1 (SIX1) expression was determined by Western blot analysis. RNA pull-down, luciferase and Western blot analysis assays were used to examine the target relationship between lncRNA CRNDE and miR-337-3p as well as between miR-337-3p and SIX1. The functional effects of lncRNA CRNDE and miR-337-3p were examined in vitro by using cell viability, colony formation, wound scratch, transwell assays, and in vivo in a xenograft tumor mouse model. RESULTS: LncRNA CRNDE was overexpressed in tumor tissues of patients with HCC. LncRNA CRNDE downregulation significantly suppressed cell proliferation and migration. Mechanistic investigations demonstrated that lncRNA CRNDE interacted with miR-337-3p and decreased its expression, thereby increasing the protein expression of miR-337-3p's target, SIX1. In addition, in vivo experiments using a xenograft tumor mouse model revealed that lncRNA CRNDE served as an oncogene, partly through sponging miR-337-3p and upregulating SIX1 in HCC. CONCLUSIONS: In this study, we report a newly identified regulatory mechanism lncRNA CRNDE/miR-337-3p/SIX1 axis, suggesting a promising therapeutic target in HCC.
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Carcinoma Hepatocelular/patologia , Proteínas de Homeodomínio/genética , Neoplasias Hepáticas/patologia , MicroRNAs/genética , RNA Longo não Codificante/genética , Animais , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/metabolismo , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Progressão da Doença , Feminino , Regulação Neoplásica da Expressão Gênica , Proteínas de Homeodomínio/metabolismo , Humanos , Neoplasias Hepáticas/genética , Masculino , Camundongos , Transplante de Neoplasias , Regulação para CimaRESUMO
BACKGROUND: To estimate the prevalence of elevated blood glucose level (EBG, including type 2 diabetes mellitus and impaired fasting glucose), and its association with non-valvular atrial fibrillation (NVAF) in Guangzhou, China. METHODS: The population-based follow-up Guangzhou Heart Study collected baseline data from July 2015 to August 2017 among 12,013 permanent residents aged > 35 from 4 Guangzhou districts. Two streets (Dadong and Baiyun) in the Yuexiu District, and one street (Xiaoguwei) and two towns (Xinzao and Nancun) in the Panyu District were chosen as representative of urban and rural areas, respectively. Each participant completed a comprehensive questionnaire, and underwent physical examination, blood sample collection for laboratory testing, electrocardiography, and other evaluations. Multivariable logistic regression analyses were used to estimate the independent association between hyperglycemia and NVAF prevalence. RESULTS: The prevalence of EBG in overall study population was 29.9%. Compared with residents without EBG, the odds ratio (OR) for AF among residents with EBG was significantly higher (1.94, 95% confidence interval [CI]: 1.40-2.70, P < 0.001), even after multivariate adjustment for metabolic abnormalities (OR = 1.60, 95% CI: 1.14-2.25, P = 0.007), and driven by women (OR = 1.80, 95% CI: 1.12-2.91, P = 0.016). CONCLUSIONS: In Guangzhou, China, prevalence of EBG is high among residents aged > 35 years and associated with a multivariate adjusted increase in prevalence of NVAF overall and in women.
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Fibrilação Atrial/epidemiologia , Glicemia/análise , Diabetes Mellitus Tipo 2/epidemiologia , Intolerância à Glucose/epidemiologia , Adulto , Fatores Etários , Idoso , Fibrilação Atrial/diagnóstico , Biomarcadores/sangue , China/epidemiologia , Estudos Transversais , Diabetes Mellitus Tipo 2/sangue , Diabetes Mellitus Tipo 2/diagnóstico , Feminino , Intolerância à Glucose/sangue , Intolerância à Glucose/diagnóstico , Humanos , Masculino , Pessoa de Meia-Idade , Prevalência , Prognóstico , Medição de Risco , Fatores de Risco , Fatores Sexuais , Fatores de Tempo , Regulação para CimaRESUMO
OBJECTIVES: The aim of the study was to examine the association of CHADS2/CHA2DS2-VASc scores with left atrial thrombus (LAT) and spontaneous echocardiographic contrast (SEC) in non-anticoagulated nonvalvular atrial fibrillation (NVAF) spontaneous patients, and to develop a new scoring system for LAT/SEC prediction. METHODS: Consecutive non-anticoagulated NVAF patients with or without LAT/SEC by transesophageal echocardiography were identified in the Guangdong General Hospital. RESULTS: Among 2,173 patients, the prevalence of LAT/SEC was 4.9%. Both predictive values of CHADS2 and CHA2DS2-VASc scores for the presence of LAT/SEC were low-to-moderate (receiver operating characteristic [ROC] = 0.591 and 0.608, respectively, p = 0.90). By multivariate analysis, non-paroxysmal AF, decreased left ventricular ejection fraction, and left atrial enlargement were positively associated with LAT/SEC, while CHADS2/CHA2DS2VASc scores were not. A new scoring system based on these 3 factors above significantly improved the discrimination for LAT/SEC (ROC = 0.792). CONCLUSIONS: CHADS2/CHA2DS2-VASc scores had limited value in predicting LAT/SEC; a new scoring system that combines AF type and echocardiographic parameters may better predict LAT/SEC as a surrogate for cardioembolic risk in NVAF patients.
Assuntos
Medição de Risco/métodos , Tromboembolia/diagnóstico por imagem , Tromboembolia/epidemiologia , Trombose/epidemiologia , Idoso , Idoso de 80 Anos ou mais , Fibrilação Atrial/complicações , Fibrilação Atrial/diagnóstico por imagem , Função do Átrio Esquerdo , China/epidemiologia , Meios de Contraste , Ecocardiografia Transesofagiana , Cardiopatias , Humanos , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Fatores de Risco , Acidente Vascular Cerebral/etiologia , Trombose/complicações , Trombose/diagnóstico por imagemRESUMO
Excessive nuclear factor κB (NF-κB) activation should be precisely controlled as it contributes to multiple immune and inflammatory diseases. However, the negative regulatory mechanisms of NF-κB activation still need to be elucidated. Various types of polyubiquitin chains have proved to be involved in the process of NF-κB activation. Many negative regulators linked to ubiquitination, such as A20 and CYLD, inhibit IκB kinase activation in the NF-κB signaling pathway. To find new NF-κB signaling regulators linked to ubiquitination, we used a small scale siRNA library against 51 ubiquitin-associated domain-containing proteins and screened out UBXN1, which contained both ubiquitin-associated and ubiquitin regulatory X (UBX) domains as a negative regulator of TNFα-triggered NF-κB activation. Overexpression of UBXN1 inhibited TNFα-triggered NF-κB activation, although knockdown of UBXN1 had the opposite effect. UBX domain-containing proteins usually act as valosin-containing protein (VCP)/p97 cofactors. However, knockdown of VCP/p97 barely affected UBXN1-mediated NF-κB inhibition. At the same time, we found that UBXN1 interacted with cellular inhibitors of apoptosis proteins (cIAPs), E3 ubiquitin ligases of RIP1 in the TNFα receptor complex. UBXN1 competitively bound to cIAP1, blocked cIAP1 recruitment to TNFR1, and sequentially inhibited RIP1 polyubiquitination in response to TNFα. Therefore, our findings demonstrate that UBXN1 is an important negative regulator of the TNFα-triggered NF-κB signaling pathway by mediating cIAP recruitment independent of VCP/p97.
Assuntos
Proteínas Adaptadoras de Transdução de Sinal/genética , Regulação da Expressão Gênica , NF-kappa B/genética , Proteínas Adaptadoras de Transdução de Sinal/antagonistas & inibidores , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Adenosina Trifosfatases/antagonistas & inibidores , Adenosina Trifosfatases/genética , Adenosina Trifosfatases/metabolismo , Proteínas de Ciclo Celular/antagonistas & inibidores , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/metabolismo , Linhagem Celular Tumoral , Genes Reporter , Células HEK293 , Células HeLa , Humanos , Proteínas Inibidoras de Apoptose/genética , Proteínas Inibidoras de Apoptose/metabolismo , Luciferases/genética , Luciferases/metabolismo , NF-kappa B/antagonistas & inibidores , NF-kappa B/metabolismo , Complexo de Proteínas Formadoras de Poros Nucleares/genética , Complexo de Proteínas Formadoras de Poros Nucleares/metabolismo , Ligação Proteica , Estrutura Terciária de Proteína , Proteólise/efeitos dos fármacos , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Proteínas de Ligação a RNA/genética , Proteínas de Ligação a RNA/metabolismo , Receptores Tipo I de Fatores de Necrose Tumoral/genética , Receptores Tipo I de Fatores de Necrose Tumoral/metabolismo , Transdução de Sinais , Bibliotecas de Moléculas Pequenas , Fator de Necrose Tumoral alfa/farmacologia , Proteína com ValosinaRESUMO
DNA damage triggers cell cycle arrest to provide a time window for DNA repair. Failure of arrest could lead to genomic instability and tumorigenesis. DNA damage-induced G1 arrest is generally achieved by the accumulation of Cyclin-dependent kinase inhibitor 1 (p21). However, p21 is degraded and does not play a role in UV-induced G1 arrest. The mechanism of UV-induced G1 arrest thus remains elusive. Here, we have identified a critical role for CUE domain-containing protein 2 (CUEDC2) in this process. CUEDC2 binds to and inhibits anaphase-promoting complex/cyclosome-Cdh1 (APC/C(Cdh1)), a critical ubiquitin ligase in G1 phase, thereby stabilizing Cyclin A and promoting G1-S transition. In response to UV irradiation, CUEDC2 undergoes ERK1/2-dependent phosphorylation and ubiquitin-dependent degradation, leading to APC/C(Cdh1)-mediated Cyclin A destruction, Cyclin-dependent kinase 2 inactivation, and G1 arrest. A nonphosphorylatable CUEDC2 mutant is resistant to UV-induced degradation. Expression of this stable mutant effectively overrides UV-induced G1-S block. These results establish CUEDC2 as an APC/C(Cdh1) inhibitor and indicate that regulated CUEDC2 degradation is critical for UV-induced G1 arrest.
Assuntos
Proteínas de Transporte/metabolismo , Pontos de Checagem da Fase G1 do Ciclo Celular/fisiologia , Proteínas de Membrana/metabolismo , Complexos Ubiquitina-Proteína Ligase/metabolismo , Proteínas Adaptadoras de Transdução de Sinal , Ciclossomo-Complexo Promotor de Anáfase , Proteínas de Transporte/antagonistas & inibidores , Proteínas de Transporte/genética , Linhagem Celular Tumoral , Ciclina A/metabolismo , Quinase 2 Dependente de Ciclina/metabolismo , Pontos de Checagem da Fase G1 do Ciclo Celular/genética , Pontos de Checagem da Fase G1 do Ciclo Celular/efeitos da radiação , Células HEK293 , Células HeLa , Humanos , Sistema de Sinalização das MAP Quinases/genética , Proteínas de Membrana/antagonistas & inibidores , Proteínas de Membrana/genética , Mutagênese Sítio-Dirigida , Fosforilação/genética , Complexos Ubiquitina-Proteína Ligase/genética , Raios UltravioletaRESUMO
This study aimed to investigate the expression of Friend leukemia virus integration 1 (Fli-1) and its correlation with the prognosis of endometrial cancer (EC). Thirty-two EC tissue samples were evaluated for Fli-1 expression using immunohistochemistry. Fli-1 showed significantly high expression in EC cells, followed by hyperplasia cells, and was negative in adjacent normal tissues. The high expression of Fli-1 was significantly associated with a high differentiation grade, mutated P53 expression, and histological subtype (p < .05). Downregulation of Fli-1 in AN3CN cells using RNA interference inhibited cell clone formation and proliferation but did not affect apoptosis and migration of the cells. This study provides the first evidence that Fli-1 expression gradually increases in parallel with disease progression, and its overexpression might predict poor prognosis in EC.