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Microbioma Gastrointestinal , Lesão Pulmonar , NF-kappa B , Transdução de Sinais , Microbioma Gastrointestinal/efeitos dos fármacos , NF-kappa B/metabolismo , Animais , Lesão Pulmonar/microbiologia , Lesão Pulmonar/metabolismo , Camundongos , Fumaça/efeitos adversos , Masculino , Camundongos Endogâmicos C57BL , Lesão por Inalação de FumaçaRESUMO
BACKGROUND: Synchronous liver metastasis (SLM) is a significant contributor to morbidity in colorectal cancer (CRC). There are no effective predictive device integration algorithms to predict adverse SLM events during the diagnosis of CRC. AIM: To explore the risk factors for SLM in CRC and construct a visual prediction model based on gray-level co-occurrence matrix (GLCM) features collected from magnetic resonance imaging (MRI). METHODS: Our study retrospectively enrolled 392 patients with CRC from Yichang Central People's Hospital from January 2015 to May 2023. Patients were randomly divided into a training and validation group (3:7). The clinical parameters and GLCM features extracted from MRI were included as candidate variables. The prediction model was constructed using a generalized linear regression model, random forest model (RFM), and artificial neural network model. Receiver operating characteristic curves and decision curves were used to evaluate the prediction model. RESULTS: Among the 392 patients, 48 had SLM (12.24%). We obtained fourteen GLCM imaging data for variable screening of SLM prediction models. Inverse difference, mean sum, sum entropy, sum variance, sum of squares, energy, and difference variance were listed as candidate variables, and the prediction efficiency (area under the curve) of the subsequent RFM in the training set and internal validation set was 0.917 [95% confidence interval (95%CI): 0.866-0.968] and 0.09 (95%CI: 0.858-0.960), respectively. CONCLUSION: A predictive model combining GLCM image features with machine learning can predict SLM in CRC. This model can assist clinicians in making timely and personalized clinical decisions.
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Millions of adenosine (A) to inosine (I) RNA editing events are reported and well-studied in eukaryotes; however, many features and functions remain unclear in prokaryotes. By combining PacBio Sequel, Illumina whole-genome sequencing, and RNA Sequencing data of two Klebsiella pneumoniae strains with different virulence, a total of 13 RNA editing events are identified. The RNA editing event of badR is focused, which shows a significant difference in editing levels in the two K. pneumoniae strains and is predicted to be a transcription factor. A hard-coded Cys is mutated on DNA to simulate the effect of complete editing of badR. Transcriptome analysis identifies the cellular quorum sensing (QS) pathway as the most dramatic change, demonstrating the dynamic regulation of RNA editing on badR related to coordinated collective behavior. Indeed, a significant difference in autoinducer 2 activity and cell growth is detected when the cells reach the stationary phase. Additionally, the mutant strain shows significantly lower virulence than the WT strain in the Galleria mellonella infection model. Furthermore, RNA editing regulation of badR is highly conserved across K. pneumoniae strains. Overall, this work provides new insights into posttranscriptional regulation in bacteria.
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Klebsiella pneumoniae , Percepção de Quorum , Virulência/genética , Klebsiella pneumoniae/genética , Percepção de Quorum/genética , Edição de RNA/genética , Sequenciamento Completo do GenomaRESUMO
Fatty acid-binding protein (Fabp)-4 is a member of the FABP family. Mammalian fabp4 has been demonstrated to involve in inflammation and immunity, whereas the related data of fish fabp4 remain limited. Therefore, we further investigated the effects of fabp4 on immunity in Ctenopharyngodon idella. The fabp4 sequence spanned 405 bp was cloned first, sharing high identity to fabp4 from other fish and mammals. Fabp4 expression was the highest in the adipose tissue, followed by the heart, muscle, and liver. In vivo, lipopolysaccharide (LPS) triggered the expression of fabp4, toll-like receptor (tlr)-22, interleukin (il)-1ß, and tumor necrosis factor (tnf)-α in the kidney and spleen. In vitro, exposing C. idella CIK cells to LPS decreased their viability, and the expression of fabp4 was also increased by LPS. However, BMS309403, an inhibitor of FABP4, mitigated these effects. Furthermore, treating the cells with LPS or fabp4 overexpression plasmids resulted in reactive oxygen species (ROS) generation and upregulation of inflammatory genes expression, including tlr22, type-I interferon (ifn-1), interferon regulatory factor (irf)-7, tnfα, il-1ß, and interferon-ß promoter stimulator 1. These effects were ameliorated by preincubation with BMS309403. Moreover, incubating the cells with glutathione reduced the production of ROS and the expression of inflammatory genes that were evoked by LPS and plasmid treatments. These results showed that fabp4 acts as a pro-inflammatory molecule via elevating ROS levels, providing a novel understanding of the molecular regulation of innate immunity in teleosts.
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Carpas , Doenças dos Peixes , Animais , Carpas/genética , Carpas/metabolismo , Proteínas de Ligação a Ácido Graxo/genética , Proteínas de Ligação a Ácido Graxo/metabolismo , Proteínas de Peixes/metabolismo , Expressão Gênica , Imunidade Inata/genética , Estresse OxidativoRESUMO
The class II trans-activator (CIITA) is the master regulator of the major histocompatibility complex (MHC) gene expression. CIITA mutations have been previously associated with several kinds of tumors, while the role of CIITA polymorphisms (rs3087456) in laryngeal squamous cell carcinoma (LSCC) is little known. We evaluate the link between CIITA polymorphisms and the existence of LSCC in patients. This study was conducted with 200 Chinese Han patients (LSCC) and 200 healthy control subjects. The association of CIITA genetic polymorphism rs3087456 with the risk of LSCC was assessed through pyrosequencing. The CIITA expression in LSCC tumor tissue and adjacent normal tissue was detected by immunohistochemistry (IHC) staining. The relationship between the genotype of rs3087456 in controls and in clinical pathology features in LSCC were analyzed, and in-silico analysis was also used for the CIITA gene. The in-silico analysis results showed that the CIITA gene is closely related to genes such as RFX5 and RFXAP. The IHC results showed that CIITA was highly expressed in LSCC tumor tissues, compared with the corresponding adjacent normal tissues. The AG, AG + AA, and A genotypes of rs3087456 of CIITA gene notably increased the risk of LSCC compared to the controls. Our study suggests that CIITA polymorphism (rs3087456) is associated with a higher risk of developing LSCC in a Chinese cohort.
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Carcinoma de Células Escamosas/genética , Predisposição Genética para Doença/genética , Proteínas Nucleares/genética , Carcinoma de Células Escamosas de Cabeça e Pescoço/genética , Transativadores/genética , Adulto , Povo Asiático , Frequência do Gene/genética , Genótipo , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
Background: NLRP3 inflammasome is an inflammatory mediator. The expression of NLRP3 inflammasome is associated with the development of various tumors and is closely related to the prognosis of tumors. However, the role of NLRP3 inflammasome in laryngeal squamous cell carcinoma (LSCC) remains unclear. This study aim to investigate the influence of NLPR3 inflammasome expression in LSCC, and especially the NLRP3 inflammasome expression level and the prognosis of LSCC after surgery in a Chinese population. Methods: We used quantitative real-time PCR and immunohistochemical (IHC) staining to calculate the mRNA (20 patients, fresh tissue) and protein expression (104 patients, paraffin tissue microarray) levels of the NLRP3 inflammasome (NLRP3/IL-18/IL-1ß/ASC/caspase-1), respectively. We also analyzed the relationship between NLRP3 inflammasome expression levels and LSCC cancer tissues compared with adjacent normal tissues and the clinical features of LSCC. Kaplan-Meier survival curves of overall survival (OS) and disease-free survival (DFS) in LSCC patients were compared and analyzed under different expression levels of the NLRP3 inflammasome. Results: Our results indicated that the mRNA expression of the NLRP3 inflammasome was higher in LSCC cancer tissues compared with adjacent normal tissues (p < 0.001). The IHC staining score also demonstrated that the expression of the NLRP3 inflammasome was higher than in the adjacent normal tissues (p < 0.001). The NLRP3 inflammasome expression also exhibited a close relationship with the clinicopathological characteristics (especially the stage of LSCC) of LSCC. Univariate Cox regression analysis and multivariate Cox regression analysis revealed that both NLRP3 and IL-1ß had an increased risk of LSCC progression (p < 0.05). The Kaplan-Meier log rank test (OS and DFS) demonstrated that high expression of NLRP3/IL-18/IL-1ß/ASC was statistically different than the low expression group (p < 0.05) of LSCC patients after surgery. Conclusion: The high expression group of the NLRP3 inflammasome (NLRP3/IL-18/IL-1ß/ASC) had a poorer prognosis (OS and DFS) than the low expression group of LSCC patients 5 years after surgery. The NLRP3 inflammasome (NLRP3/IL-18/IL-1ß/ASC) may be used as an auxiliary indicator to predict LSCC patient prognosis after surgery.
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Reactive oxygen speciesinduced cyclophilin A (CyPA) release from vascular smooth muscle cells (VSMCs) may be inhibited by simvastatin in vitro. The present study aimed to further examine the effect of simvastatin on serum CyPA levels and the basigin (CD147)extracellular signalregulated kinase (ERK) 1/2cyclin pathway during thoracic aorta remodeling. The mechanisms through which simvastatin may inhibit CyPA secretion from VSMCs were further investigated. Serum CyPA levels and the expression kinetics of CyPAassociated signaling pathways were examined following simvastatin treatment in rat thoracic aortas during hypertension. Cell lysates were prepared from middle layer of thoracic aortas at 1, 4, 8 and 12 weeks subsequent to surgery. ELISA analysis revealed that serum CyPA levels were gradually increased with the progression of thoracic aorta remodeling. Western blotting demonstrated that the expression of CD147, phosphorylatedERK1/2, cyclin D1, cyclin A, and cyclin E were increased with the progression of thoracic aorta remodeling. Simvastatin administration for 4, 8 and 12 weeks diminished all these changes, as observed in the hypertensive group. VSMCs from simvastatintreated rats secreted a decreased amount of CyPA compared with VSMCs from hypertensive rats. In addition, pretreatment with geranylgeraniol partly reversed the inhibitory effect of simvastatin on LY83583induced CyPA secretion in cultured VSMCs, whereas GGTI298 and KD025 [a selective Rhoassociated protein kinase 2 (ROCK2) inhibitor] mimicked the inhibitory effect of simvastatin. The present study demonstrated that simvastatin alleviated thoracic aorta remodeling by reducing CyPA secretion and expression of the CD147ERK1/2cyclin signaling pathway. In addition, the results of the present study demonstrated that the RhoROCK2 pathway mediated CyPA secretion from VSMCs.
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Aorta Torácica/metabolismo , Aorta Torácica/patologia , Ciclofilina A/metabolismo , Transdução de Sinais/efeitos dos fármacos , Sinvastatina/farmacologia , Remodelação Vascular/efeitos dos fármacos , Quinases Associadas a rho/metabolismo , Animais , Basigina/metabolismo , Biomarcadores , Biópsia , Ciclinas/metabolismo , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Masculino , RatosRESUMO
The present study was aimed to investigate the role of necroptosis in the pathogenesis of acute respiratory distress syndrome (ARDS). The rat model of ARDS was induced by intravenous injection of oleic acid (OA), and observed for 4 h. The lung injury was evaluated by arterial blood gas, lung wet-dry weight ratio (W/D) and histological analyses. Simultaneously, bronchoalveolar lavage fluid (BALF) was collected for total and differential cell analysis and total protein determination. Tumor necrosis factor alpha (TNF-α) level in BALF was determined with a rat TNF-α ELISA kit. Expressions of receptor interacting protein kinase 1 (RIPK1), RIPK3 and mixed lineage kinase domain-like protein (MLKL) in lung tissue were determined by Western blot and immunohistochemical staining. The interaction between RIPK1 and RIPK3 was explored by immunoprecipitation. The results showed that, compared with those in control group, total white blood cells count (WBC), polymorphonuclear percentage (PMN%), total protein concentration, TNF-α level in BALF, W/D, and the alveolar-arterial oxygen tension difference (P(A-a)O2) in OA group were significantly increased at 4 h after OA injection. Western blot and immunostaining further showed remarkably increased expressions of RIPK1, RIPK3 and MLKL in lung tissue from OA group. Additionally, immunoprecipitation results indicated an enforced interaction between RIPK1 and RIPK3 in OA group. Collectively, the TNF-α level in BALF and the RIPK1-RIPK3-MLKL signaling pathway in lung tissue were found to be upregulated and activated with the process of ARDS. These findings implicate that RIPK1/RIPK3-mediated necroptosis plays a possible role in the pathogenesis of ARDS, which may provide a new idea to develop novel drugs for the therapy of ARDS.
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Pneumopatias , Transtornos Respiratórios , Doença Aguda , Animais , Líquido da Lavagem Broncoalveolar , Modelos Animais de Doenças , Necrose , Ácido Oleico , Ratos , Proteína Serina-Treonina Quinases de Interação com Receptores , Transdução de Sinais , Fator de Necrose Tumoral alfaRESUMO
Necroptosis is a recently discovered necrotic cell death which is regulated by receptor interacting protein kinase 1 (RIPK1) and RIPK3 under the stimulus of death signal and can be inhibited by necrostatin-1 (Nec-1) specifically. Therefore, the aim was to investigate the role of necroptosis in a rat model of acute respiratory distress syndrome (ARDS) induced by oleic acid (OA) and assess the effect of Nec-1 on lung injury in ARDS. Our results found that RIPK1, RIPK3 and mixed lineage kinase domain-like protein (MLKL) were abundantly expressed in rat lung tissues of OA-induced ARDS. Nec-1 pretreatment improved pulmonary function and attenuated lung edema dramatically in OA-induced ARDS rats. Furthermore, Nec-1 reduced RIPK1-RIPK3 interaction and down-regulated RIPK1-RIPK3-MLKL signal pathway, and inhibited inflammatory response by reducing neutrophil infiltration and protein leakage into lung tissue in OA-induced ARDS. Collectively, our study proves the intervention of necroptosis in OA-induced ARDS. Moreover, our findings imply that Nec-1 plays an important role in the treatment of ARDS via inhibiting necroptosis and inflammation.
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Imidazóis/uso terapêutico , Indóis/uso terapêutico , Substâncias Protetoras/uso terapêutico , Síndrome do Desconforto Respiratório/tratamento farmacológico , Síndrome do Desconforto Respiratório/prevenção & controle , Animais , Apoptose/efeitos dos fármacos , Líquido da Lavagem Broncoalveolar , Agregação Celular/efeitos dos fármacos , Contagem de Células , Modelos Animais de Doenças , Imidazóis/farmacologia , Imuno-Histoquímica , Indóis/farmacologia , Inflamação/patologia , Masculino , Necrose , Ácido Oleico , Substâncias Protetoras/farmacologia , Ratos Sprague-Dawley , Síndrome do Desconforto Respiratório/induzido quimicamente , Síndrome do Desconforto Respiratório/fisiopatologia , Transdução de Sinais/efeitos dos fármacos , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Posttraumatic nightmares are a core component of posttraumatic stress disorder (PTSD) and mechanistically linked to the development and maintenance of this disorder, but little is known about their mechanism. We utilized a communication box to establish an animal model of physiological stress (foot-shock [FS]) and psychological stress (PS) to mimic the direct suffering and witnessing of traumatic events. Twenty-one days after traumatic stress, some of the experimental animals presented startled awakening (i.e., were startled awake by a supposed "nightmare") with different electroencephalographic spectra features. Our neuroanatomical results showed that the secondary somatosensory cortex and primary auditory cortex may play an important role in remote traumatic memory retrieval in FS "nightmare" (FSN) rats, whereas the temporal association cortex may play an important role in PS "nightmare" (PSN) rats. The FSN and PSN groups possessed common emotion evocation circuits, including activation of the amygdala and inactivation of the infralimbic prefrontal cortex and ventral anterior cingulate cortex. The decreased activity of the granular and dysgranular insular cortex was only observed in PSN rats. The present results imply that different types of stress may cause PTSD-like "nightmares" in rodents and identified the possible neurocircuitry of memory retrieval and emotion evocation.
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Córtex Auditivo/fisiologia , Sonhos/fisiologia , Proteínas Proto-Oncogênicas c-fos/metabolismo , Córtex Somatossensorial/fisiologia , Transtornos de Estresse Pós-Traumáticos/fisiopatologia , Estresse Fisiológico/fisiologia , Estresse Psicológico/fisiopatologia , Animais , Eletroencefalografia , Feminino , Memória/fisiologia , Modelos Animais , Ratos , Ratos Sprague-Dawley , Sono/fisiologia , Lobo TemporalRESUMO
Echinomycin, a member of the quinoxaline family of antibiotics, is known to be a small-molecule inhibitor of hypoxia inducible factor-1 (HIF-1) DNA binding activity. Recently, it has been shown to suppress mammalian target of rapamycin (mTOR) signaling and growth in leukemia cell lines. In this study, we investigated whether echinomycin interacts with the FKBP12 protein. Molecular docking was used, and the predicted binding energy was -10.61 kcal/mol. Moreover, surface plasmon resonance imaging and fluorescence quenching techniques were used to validate this interaction. Echinomycin binds to FKBP12 with a strong binding affinity comparable with rapamycin. Furthermore, the echinomycin-FKBP12 complex has been shown to affect calcineurin activity when tested in a calcineurin phosphatase inhibition assay. All of these studies have shown that echinomycin may have a double impact on HIF signaling by direct inhibition and through mTOR.
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Calcineurina/metabolismo , Equinomicina/metabolismo , Equinomicina/farmacologia , Proteína 1A de Ligação a Tacrolimo/metabolismo , Calcineurina/química , Equinomicina/química , Ativação Enzimática/efeitos dos fármacos , Modelos Biológicos , Modelos Moleculares , Conformação Molecular , Simulação de Acoplamento Molecular , Monoéster Fosfórico Hidrolases/metabolismo , Ligação Proteica , Reprodutibilidade dos Testes , Transdução de Sinais/efeitos dos fármacos , Ressonância de Plasmônio de Superfície/métodosRESUMO
Endothelial progenitor cells (EPCs) play a key role in tissue repair and regeneration. Previous studies have shown a positive correlation between the number of circulating EPCs and clinical outcomes of patients with traumatic brain injury (TBI). A recent study has further shown that intravenous infusion of human umbilical cord blood-derived endothelial colony-forming cells (ECFCs) improves outcomes of mice subjected to experimental TBI. This follow-up study was designed to determine whether intracerebroventricular (i.c.v.) infusion of ECFCs, which may reduce systemic effects of these cells, could repair the blood-brain barrier (BBB) and promote angiogenesis of mice with TBI. Adult nude mice were exposed to fluid percussion injury and transplanted i.c.v. with ECFCs on day 1 post-TBI. These ECFCs were detected at the TBI zone 3 days after transplantation by SP-DiIC18(3) and fluorescence in situ hybridization. Mice with ECFCs transplant had reduced Evans blue extravasation and brain water content, increased expression of ZO-1 and claudin-5, and showed a higher expression of angiopoietin 1. Consistent with the previous report, mice with ECFCs transplant had also increased microvascular density. Modified neurological severity score and Morris water maze test indicated significant improvements in motor ability, spatial acquisition and reference memory in mice receiving ECFCs, compared to those receiving saline. These data demonstrate the beneficial effects of ECFC transplant on BBB integrity and angiogenesis in mice with TBI.
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Barreira Hematoencefálica/fisiologia , Lesões Encefálicas/cirurgia , Transplante de Células-Tronco de Sangue do Cordão Umbilical/métodos , Células Endoteliais/fisiologia , Células Endoteliais/transplante , Neovascularização Fisiológica/fisiologia , Animais , Barreira Hematoencefálica/cirurgia , Lesões Encefálicas/patologia , Células Cultivadas , Feminino , Células-Tronco Hematopoéticas/fisiologia , Humanos , Infusões Intraventriculares , Camundongos , Camundongos Endogâmicos BALB C , Camundongos NusRESUMO
OBJECTIVE: To observe the influence of electroacupuncture (EA) of "Chize" (LU 5, He-sea acupoint) and "Shangjuxu" (ST 37, lower He-sea acupoint) on mesenteric microcirculation, vasoactive intestinal peptide (VIP) levels in the lung, colon and hypothalamus tissues in rats with chronic obstructive pulmonary disease (COPD), so as to investigate its mechanism underlying improvement of lung derived intestinal disorders in clinical practice. METHODS: Thirty-two male Wistar rats were randomly divided into normal control, model, EA-Chize (LU 5, EA-LU 5) and EA-Shangjuxu (ST 37, EA-ST 37) groups, with 8 rats being in each group. COPD model was established by intratracheal infusion of Lipopolysaccharide (LPS, 1 mg/mL, 0.2 mL/rat) and forced inhaling smoke, once daily for 28 days. EA was applied to bilateral LU 5 and ST 37 for 20 min, once every other day for 12 sessions. The state of mesenteric microcirculation was observed under microscope and divided into grade 0 (stagnation of blood flow), I (slow flowing and silt-like state), II (faster flowing with slight or obvious grainy feeling) and III (fast flowing without grainy feeling). The contents of VIP in the lung, colon and hypothalamus were detected using radioimmunoassay (RIA). RESULTS: Following modeling, the microvascular calibers were increased slightly in the model, EA-LU 5 and EA-ST 37 groups. Compared with the normal group, the blood flow velocity was increased significantly in model group (P < 0.05). In comparison with the model group, the blood flow velocity was reduced significantly in EA-LU 5 and EA-ST 37 groups (P < 0.01). There were no significant differences between EA-LU 5 and EA-ST 37 groups in blood flow velocity, among the four groups in VIP contents of the lung tissue (P > 0.05). The content of VIP in the colon was markedly higher in the model group than in the normal group, and that in the hypothalamus was obviously lower in the EA-LU 5 group than in the model group (all P < 0.05). CONCLUSION: EA stimulation of "Chize" (LU 5) can notably reduce hypothalamic VIP content and slow down blood flow velocity of the mesenteric microvessels in COPD rats.