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Switching of vascular smooth muscle cells (VSMCs) from a contractile phenotype to an adverse proliferative phenotype is a hallmark of atherosclerosis or vascular restenosis. However, the genetic modulators responsible for this switch have not been fully elucidated in humans nor have they been correlated with clinical abnormalities. This study investigated genetic mechanisms involved in phenotypic switching of VSMCs at non-defect areas of the aorta in patients with atherosclerosis. Aortic wall samples were obtained from patients with (N = 53) and without (N = 27) atherosclerosis undergoing cardiovascular surgery. Vascular smooth muscle cell cultures were generated, and expression of microRNA-145 (miR-145), its target gene Kruppel-Like Factor 5 (KLF5) and Myocardin (MYOCD, a smooth muscle-specific transcriptional coactivator) were analysed using RT-qPCR, along with expression of relevant proteins. Vascular smooth muscle cells were transduced with miR-145 inhibitor and mimic to determine the effect of miR-145 expression on VSMC proliferation. miR-145 expression decreased while KLF5 expression increased in atherosclerotic aortas. Atherosclerotic samples and VSMCs had decreased expression of contractile markers calponin and alpha smooth muscle actin (α-SMA) and MYOCD. miR-145 inhibitor-transduced VSMCs from non-atherosclerotic patients showed decreased expression of calponin and α-SMA and increased proliferation compared with non-transduced controls, and these levels were close to those of atherosclerotic patients. miR-145 mimic-transduced VSMCs from atherosclerotic patients showed increased expression of calponin and α-SMA and decreased proliferation compared with non-transduced controls, and these levels were close to those found in non-atherosclerotic patients. These data demonstrate that miR-145 modulates the phenotypic switch of VSMCs from a contractile to a proliferative state via KLF5 and MYOCD in atherosclerosis.
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Aorta/patología , Aterosclerosis/genética , Aterosclerosis/patología , MicroARNs/metabolismo , Músculo Liso Vascular/patología , Miocitos del Músculo Liso/metabolismo , Adulto , Biomarcadores/metabolismo , Proliferación Celular , Femenino , Regulación de la Expresión Génica , Humanos , Masculino , MicroARNs/genética , Persona de Mediana Edad , Especificidad de Órganos , FenotipoRESUMEN
Sirtuin3 (SIRT3) is an important member of the sirtuin family of protein deacetylases that is localized to mitochondria and linked to lifespan extension in organisms ranging from yeast to humans. As aged cells have less regenerative capacity and are more susceptible to oxidative stress, we investigated the effect of ageing on SIRT3 levels and its correlation with antioxidant enzyme activities. Here, we show that severe oxidative stress reduces SIRT3 levels in young human mesenchymal stromal/stem cells (hMSCs). Overexpression of SIRT3 improved hMSCs resistance to the detrimental effects of oxidative stress. By activating manganese superoxide dismutase (MnSOD) and catalase (CAT), SIRT3 protects hMSCs from apoptosis under stress. SIRT3 expression, levels of MnSOD and CAT, as well as cell survival showed little difference in old versus young hMSCs under normal growth conditions, whereas older cells had a significantly reduced capacity to withstand oxidative stress compared to their younger counterparts. Expression of the short 28 kD SIRT3 isoform was higher, while the long 44 kD isoform expression was lower in young myocardial tissues compared with older ones. These results suggest that the active short isoform of SIRT3 protects hMSCs from oxidative injury by increasing the expression and activity of antioxidant enzymes. The expression of this short isoform decreases in cardiac tissue during ageing, leading to a reduced capacity for the heart to withstand oxidative stress.
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Apoptosis/genética , Células Madre Mesenquimatosas/metabolismo , Estrés Oxidativo/genética , Sirtuina 3/genética , Envejecimiento , Antioxidantes/metabolismo , Catalasa/genética , Línea Celular , Regulación de la Expresión Génica , Humanos , Células Madre Mesenquimatosas/patología , Especies Reactivas de Oxígeno/metabolismo , Sirtuina 3/biosíntesis , Superóxido Dismutasa/genéticaRESUMEN
BACKGROUND: End-to-side anastomoses to connect the distal end of the great saphenous vein (GSV) to small target coronary arteries are commonly performed in sequential coronary artery bypass grafting (CABG). However, the oversize diameter ratio between the GSV and small target vessels at end-to-side anastomoses might induce adverse hemodynamic condition. The purpose of this study was to describe a distal end side-to-side anastomosis technique and retrospectively compare the effect of distal end side-to-side versus end-to-side anastomosis on graft flow characteristics. METHODS: We performed side-to-side anastomoses to connect the distal end of the GSV to small target vessels on 30 patients undergoing off-pump sequential CABG in our hospital between October 2012 and July 2013. Among the 30 patients, end-to-side anastomoses at the distal end of the GSV were initially performed on 14 patients; however, due to poor graft flow, those anastomoses were revised into side-to-side anastomoses. We retrospectively compared the intraoperative graft flow characteristics of the end-to-side versus side-to-side anastomoses in the 14 patients. The patient outcomes were also evaluated. RESULTS: We found that the side-to-side anastomosis reconstruction improved intraoperative flow and reduced pulsatility index in all the 14 patients significantly. The 16 patients who had the distal end side-to-side anastomoses performed directly also exhibited satisfactory intraoperative graft flow. Three-month postoperative outcomes for all the patients were satisfactory. CONCLUSIONS: Side-to-side anastomosis at the distal end of sequential vein grafts might be a promising strategy to connect small target coronary arteries to the GSV.
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Puente de Arteria Coronaria Off-Pump/métodos , Enfermedad de la Arteria Coronaria/cirugía , Circulación Coronaria , Vasos Coronarios/cirugía , Vena Safena/cirugía , Anciano , Anastomosis Quirúrgica , Enfermedad de la Arteria Coronaria/diagnóstico , Enfermedad de la Arteria Coronaria/fisiopatología , Vasos Coronarios/fisiopatología , Femenino , Hemodinámica , Humanos , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Vena Safena/fisiopatología , Factores de Tiempo , Resultado del TratamientoRESUMEN
BACKGROUND: For complete revascularization, patients with diffuse coronary artery disease should have a coronary endarterectomy and a coronary artery bypass graft (CE-CABG). Sadly, CE can lead to a lack of endothelium, which raises the risk of thrombotic events. Even though daily dual antiplatelet therapies (DAPT) have been shown to reduce thrombotic events, the risk of perioperative thrombotic events is high during the high-risk period after CE-CABG, and there is no consistent protocol to bridge DAPT. This trial aims to compare safety and efficacy between tirofiban and heparin as DAPT bridging strategies after CE-CABG. METHODS: In phase I, 266 patients undergoing CE-CABG will be randomly assigned to tirofiban and heparin treatment groups to compare the two treatments in terms of the primary safety endpoint, chest tube drainage in the first 24 h. If the phase I trial shows tirofiban non-inferiority, phase II will commence, in which an additional 464 patients will be randomly assigned. All 730 patients will be studied to compare major cardiovascular and cerebrovascular events (MACCEs) between the groups in the first 30 days after surgery. DISCUSSION: Given the possible benefits of tirofiban administration after CE-CABG, this trial has the potential to advance the field of adult coronary heart surgery. TRIAL REGISTRATION: chictr.org.cn, ChiCTR2200055697. Registered 6 January 2022. https://www.chictr.org.cn/com/25/showproj.aspx?proj=149451 . Current version: 20,220,620.
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Enfermedad de la Arteria Coronaria , Inhibidores de Agregación Plaquetaria , Adulto , Humanos , Tirofibán/efectos adversos , Inhibidores de Agregación Plaquetaria/efectos adversos , Heparina/efectos adversos , Resultado del Tratamiento , Puente de Arteria Coronaria/efectos adversos , Puente de Arteria Coronaria/métodos , Enfermedad de la Arteria Coronaria/cirugía , Endarterectomía , Fibrinolíticos/uso terapéutico , Ensayos Clínicos Controlados Aleatorios como Asunto , Estudios Multicéntricos como AsuntoRESUMEN
Atherosclerosis is the most notable cardiovascular disease, the latter being the main cause of death globally. Endothelial cell dysfunction plays a major role in the pathogenesis of atherosclerosis. However, it is currently unclear which genes are involved between endothelial cell dysfunction and atherosclerosis. This study was aimed at identifying these genes. Based on the GSE83500 dataset, the quantification of endothelial cell function was conducted using single-sample gene set enrichment analysis; the coexpression modules were conducted using weighted correlation network analysis. After building module-trait relationships, tan and yellow modules were regarded as hub modules. 10 hub genes from each hub module were identified by the protein-protein interaction network analysis. The key genes (RAB5A, CTTN, ITGB1, and MMP9) were obtained by comparing the expression differences of the hub gene between atherosclerotic and normal groups from the GSE28829 and GSE43292 datasets, respectively. ROC analysis showed the diagnostic value of key genes. Moreover, the differential expression of key genes in normal and atherosclerotic aortic walls was verified. In vitro, we establish a model of ox-LDL-injured endothelial cells and transfect RAB5A overexpression and shRNA plasmids. The results showed that overexpression of RAB5A ameliorates the proliferation and migration function of ox-LDL-injured endothelial cells, including the ability of tubule formation. It was speculated that the interferon response, Notch signaling pathways, etc. were involved in this function of RAB5A by using gene set variation analysis. With the multiple bioinformatics analysis methods, we detected that yellow and tan modules are related to the abnormal proliferation and migration of endothelial cells associated with atherosclerosis. RAB5A, CTTN, ITGB1, and MMP9 can be used as potential targets for therapy and diagnostic markers. In vitro, overexpression of RAB5A can ameliorate the proliferation and migration function of ox-LDL-injured endothelial cells, and the possible molecules involved in this process were speculated.
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Aterosclerosis , Bases de Datos Genéticas , Células Endoteliales/metabolismo , Regulación de la Expresión Génica , Aterosclerosis/genética , Aterosclerosis/metabolismo , Línea Celular , Biología Computacional , HumanosRESUMEN
This review highlights vital details that can be easily overlooked and discuss how to identify and fix failed cannulation from another novel insight. Appropriate arterial cannulation strategy during cardiopulmonary bypass (CPB) in Stanford type A aortic dissection (AAD) is highly necessary to reach satisfactory perfusion effects and appreciable clinical outcomes. Despite several previously published reviews on cannulation strategies in AAD, most focus on the advantages and disadvantages by comparing various cannulation strategies. In fact, most of evidence came from retrospective studies. More importantly, however, some important details and novel approaches maybe overlooked due to variety reasons. These overlooked details also make sense in clinical practice. Papers related to cannulation refer to type AAD were retrieved and analyzed from the PubMed and Medline database. The key words such as "aortic dissection", "cannula", "cannulation", "cannulation strategy", "cerebral perfusion", "type I aortic dissection" were conducted and analyzed. In addition, we looked at some new and very significant specific perfusion techniques such as anterograde cerebral perfusion combined with retrograde inferior vena caval perfusion (RIVP) and reperfusion via the right carotid artery before surgery. The arterial cannulation site and strategy should be determined individually. Monitoring measures are very necessary in the whole procedure.
RESUMEN
Engineered heart tissues (EHTs) are regarded as being the most promising alternative to synthetic materials, and autologous mesenchymal stem cells (MSCs) are widely used as seeding cells. However, few studies have evaluated the feasibility of using MSCs from patients with cyanotic congenital heart disease (C-CHD) as seeding cells for EHTs, in comparison with cells from patients of acyanotic congenital heart disease (A-CHD). In the present study, we cultured MSCs from A-CHD and C-CHD patients in normoxia or hypoxia conditions, and compared their pro-angiogenic, anti-apoptotic and inflammation-modulatory potentials. In vivo, we seeded the cells into collagen patches conjugated with, or without, proangiogenic cytokines, which were used to repair the right ventricular outflow tract (RVOT) of rats. The in vitro results showed that C-CHD MSCs expressed higher levels of VEGFA and VEGFR2, and secreted more pro-angiogenic and anti-inflammatory cytokines under hypoxic conditions. On the other hand, apoptosis-related genes from C-CHD MSCs were modulated adaptably, converting these cells into an anti-apoptotic phenotype. In vivo studies demonstrated that in 4 weeks after RVOT reconstruction, cytokine-immobilized patches seeded with C-CHD MSCs exhibited preserved morphology, prolonged cell survival and enhanced angiogenesis compared to A-CHD MSCs. C-CHD MSCs that undergo "naturally hypoxic precondition" present a better cell source for EHTs, which would provide a promising individualized biomaterial for C-CHD patients.
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Cardiopatías Congénitas , Células Madre Mesenquimatosas , Ingeniería de Tejidos , Animales , Células Cultivadas , Corazón , Cardiopatías Congénitas/terapia , Humanos , Hipoxia , RatasRESUMEN
Adiposederived stem cells (ADSCs) and bone marrowderived stem cells (BMSCs) are considered to be prospective sources of mesenchymal stromal cells (MSCs), that can be used in cell therapy for atherosclerosis. The present study investigated whether ADSCs cocultured with M1 foam macrophages via treatment with oxidized lowdensity lipoprotein (oxLDL) would lead to similar or improved antiinflammatory effects compared with BMSCs. ADSCs, peripheral blood monocytes, BMSCs and oxLDL were isolated from ten coronary heart disease (CHD) patients. After three passages, the supernatants of the ADSCs and BMSCs were collected and systematically analysed by liquid chromatographyquadrupole timeofflightmass spectrometry (6530; Agilent Technologies, Inc., Santa Clara, CA, USA). Cis9, trans11 was deemed to be responsible for the potential differences in the metabolic characteristics of ADSCs and BMSCs. These peripheral blood monocytes were characterized using flow cytometry. Following peripheral blood monocytes differentiation into M1 macrophages, the formation of M1 foam macrophages was achieved through treatment with oxLDL. Overall, 2x106 ADSCs, BMSCs or BMSCs+cis9, trans11 were cocultured with M1 foam macrophages. Antiinflammatory capability, phagocytic activity, antiapoptotic capability and cell viability assays were compared among these groups. It was demonstrated that the accumulation of lipid droplets decreased following ADSCs, BMSCs or BMSCs+cis9, trans11 treatment in M1 macrophages derived from foam cells. Consistently, ADSCs exhibited great advantageous antiinflammatory capabilities, phagocytic activity, antiapoptotic capability activity and cell viability over BMSCs or BMSCs+cis9, trans11. Additionally, BMSCs+cis9, trans11 also demonstrated marked improvement in antiinflammatory capability, phagocytic activity, antiapoptotic capability activity and cell viability in comparison with BMSCs. The present results indicated that ADSCs would be more appropriate for transplantation to treat atherosclerosis than BMSCs alone or BMSCs+cis9, trans11. This may be an important mechanism to regulate macrophage immune function.
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Tejido Adiposo/citología , Células de la Médula Ósea/metabolismo , Células Espumosas/metabolismo , Inflamación/etiología , Inflamación/metabolismo , Lipoproteínas LDL/efectos adversos , Células Madre Mesenquimatosas/metabolismo , Anciano , Apoptosis , Células de la Médula Ósea/citología , Supervivencia Celular , Citocinas/metabolismo , Femenino , Células Espumosas/citología , Expresión Génica , Perfilación de la Expresión Génica , Humanos , Inflamación/patología , Mediadores de Inflamación/metabolismo , Metabolismo de los Lípidos , Macrófagos/metabolismo , Masculino , Células Madre Mesenquimatosas/citología , Metaboloma , Metabolómica/métodos , Persona de Mediana EdadRESUMEN
Surgical ventricular reconstruction (SVR) can restore cardiac function for left ventricular aneurysm to some extent. However, the patches used in this treatment have some limitations such as stiffness and calcification. Engineering heart tissues (EHTs) have emerged as a promising biomaterial to repair damaged heart. Nevertheless, selecting optimal candidate cells for EHTs has been controversial. Aging is a major consideration for seed cells derived from elderly patients. Hence, this study was aimed to assess the proliferation of, antiapoptosis potential of, and expression of senescence-associated factors (eg, SA-ß-Gal, cyclin-dependent kinase inhibitor 2A (P16), cyclin-dependent kinase inhibitor 1 (P21) in adipose-derived stem cells (ADSCs), and bone marrow stem cells (BMSCs) in vitro. In addition, cardiac function, cell survival, and angiogenesis of ADSCs and BMSCs after SVR were assessed in vivo. The in vitro results showed that old ADSCs (OAs) grew faster; expressed lower levels of SA-ß-Gal, P16, and P21; and possessed more pronounced antiapoptosis activity than old BMSCs (OBs). The in vivo results demonstrated that 28 days after patch implantation, animals that received OAs patches showed better restoration of cardiac function than animals that received OBs patches. Meanwhile, old ADSCs possessed more potential regarding cell survival and angiogenesis. These results suggest that ADSCs may be superior to BMSCs with regard to autologous cell transplantation in elderly patients.
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Tejido Adiposo/citología , Células de la Médula Ósea/citología , Senescencia Celular , Ventrículos Cardíacos/citología , Células Madre Mesenquimatosas/citología , Donantes de Tejidos , Animales , Apoptosis , Biomarcadores/metabolismo , Células de la Médula Ósea/metabolismo , Proliferación Celular , Forma de la Célula , Supervivencia Celular , Humanos , Cinética , Masculino , Células Madre Mesenquimatosas/metabolismo , Persona de Mediana Edad , Neovascularización Fisiológica , Ratas Sprague-Dawley , Adulto JovenRESUMEN
BACKGROUND: Engineered heart tissues (EHTs) present a promising alternative to current materials for surgical ventricular restoration (SVR); however, the clinical application remains limited by inadequate vascularization postimplantation. Moreover, a suitable and economic animal model for primary screening is another important issue. METHODS: Recently, we used 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride chemistry (EDC) to initiate a strengthened, cytokine-conjugated collagenous platform with a controlled degradation speed. In vitro, the biomaterial exhibited an enhanced mechanical strength maintaining a porous ultrastructure, and the constant release of cytokines promoted the proliferation of seeded human mesenchymal stem cells (hMSCs). In vivo, with the hMSC-seeded, cytokine-immobilized patch (MSCs + GF patch), we performed modified SVR for rats with left ventricular aneurysm postmyocardial infarction (MI). Overall, the rats that underwent modified SVR lost less blood and had lower mortality. After 4 weeks, the rats repaired with this cell-seeded, cytokine-immobilized patch presented preserved cardiac function, beneficial morphology, enhanced cell infiltration, and functional vessel formation compared with the cytokine-free (MSC patch), cell-free (GF patch), or blank controls (EDC patch). Furthermore, the degradable period of the collagen patch in vivo extended up to 3 months after EDC treatment. CONCLUSIONS: EDC may substantially modify collagen scaffold and provide a promising and practical biomaterial for SVR.
RESUMEN
Aberrant proliferation and migration of vascular smooth muscle cells (VSMCs) play a crucial role in the pathogenesis of cardiovascular diseases including coronary heart disease, restenosis and atherosclerosis. MicroRNAs are a class of small, non-coding and endogenous RNAs that play critical roles in VSMCs function. In this study, we showed that PDGF-bb, as a stimulant, promoted VSMCs proliferation and suppressed the expression of miR-599. Moreover, overexpression of miR-599 inhibited VSMCs proliferation and also suppressed the PCNA and ki-67 expression. In addition, we demonstrated that ectopic expression of miR-599 repressed the VSMCs migration. We also showed that miR-599 inhibited type I collagen, type V collagen and proteoglycan expression. Furthermore, we identified TGFb2 as a direct target gene of miR-599 in VSMCs. Overexpression of TGFb2 reversed miR-599-induced inhibition of VSMCs proliferation and type I collagen, type V collagen and proteoglycan expression. In conclusion, our findings suggest miR-599 plays a crucial role in controlling VSMCs proliferation and matrix gene expression by regulating TGFb2 expression.
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Movimiento Celular/genética , Proliferación Celular/genética , MicroARNs/genética , Músculo Liso Vascular/citología , Proteínas Proto-Oncogénicas c-sis/metabolismo , Factor de Crecimiento Transformador beta2/genética , Aterosclerosis/genética , Aterosclerosis/patología , Becaplermina , Línea Celular , Colágeno Tipo I/biosíntesis , Colágeno Tipo V/biosíntesis , Enfermedad Coronaria/genética , Enfermedad Coronaria/patología , Reestenosis Coronaria/genética , Reestenosis Coronaria/patología , Humanos , MicroARNs/biosíntesis , Proteoglicanos/biosíntesis , Factor de Crecimiento Transformador beta2/biosíntesisRESUMEN
Aberrant proliferation of vascular smooth muscle cells [VSMCs] is implicated in the pathogenesis of vascular pathologies such as atherosclerosis and restenosis. Accumulating evidences have revealed that microRNAs are involved in cell proliferation in various pathological conditions. In the present study, we showed that miR-136 was up regulated in human coronary atherosclerotic plaques when compared with normal coronary artery tissues. Moreover, miR-136 levels were up regulated in proliferative vascular smooth muscle cells induced by platelet-derived growth factor [PDGF] or serum. In cultured VSMCs, over expression of miR-136 stimulated cell proliferation. PPP2R2A was proved to be the direct target gene of miR-136 and knockdown of PPP2R2A had a proliferative effect on VSMCs. miR-136-induced PPP2R2A down-regulation was accompanied by increased expression of ERK1/2 phosphorylation. Inhibition of ERK1/2 abolished the effect of miR-136 and knockdown of PPP2R2A on VSMCs proliferation. In summary, aberrant miR-136 up regulation in atherosclerosis contributes to abnormal VSMC proliferation through suppressing the ERK1/2 pathway by targeting PPP2R2A. Our study also suggested that specific modulation of miR-136 in human VSMCs may provide a potential approach for the treatment of atherosclerosis.
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Aterosclerosis/tratamiento farmacológico , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , MicroARNs/farmacología , Músculo Liso Vascular/citología , Músculo Liso Vascular/efectos de los fármacos , Proteína Fosfatasa 2/efectos de los fármacos , Proliferación Celular , Enfermedad de la Arteria Coronaria/tratamiento farmacológico , Enfermedad de la Arteria Coronaria/patología , Técnicas de Silenciamiento del Gen , Humanos , MicroARNs/uso terapéutico , Factor de Crecimiento Derivado de Plaquetas/farmacología , Biosíntesis de Proteínas/efectos de los fármacos , Proteína Fosfatasa 2/biosíntesis , Proteína Fosfatasa 2/genética , Sincalida/metabolismoRESUMEN
Pulmonary dysfunction is one of the most frequent complications associated with cardiopulmonary bypass (CPB). Multiple factors, including the contact of blood with the artificial surface of the CPB circuit, ischemiareperfusion and lung ventilator arrest elicit inflammatory reactions, consequently resulting in CPBinduced lung injury. The proinflammatory cytokine tumor necrosis factorα (TNFα) has been demonstrated to have a critical role in mediating CPBinduced pulmonary inflammation. The present review evaluated previous studies and summarized the effects of CPB on TNFα level in the serum and lung tissue of patients and animal models of CPB, the underlying mechanism of TNFαmediated lung injury and the therapeutic strategies for the inhibition of TNFα activity and production to attenuate CPBinduced lung injury. TNFα level in the serum and lung tissue is significantly increased during and following CPB. TNFα mediates CPBinduced lung damage by directly inducing apoptosis in alveolar epithelial cells and lung endothelial cells and by indirectly modulating the function of immune cells, including monocytes and macrophages. A functional neutralizing antibody to TNFα can reduce pulmonary TNFα production and attenuate CPBinduced lung injury in a rabbit model of CPB. Inhibition of TNFα function and production using a neutralizing antibody to TNFα appears to be a promising therapeutic strategy to alleviate CPBinduced lung injury.
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Antiinflamatorios/uso terapéutico , Puente Cardiopulmonar/efectos adversos , Lesión Pulmonar/tratamiento farmacológico , Lesión Pulmonar/etiología , Factor de Necrosis Tumoral alfa/antagonistas & inhibidores , Animales , Antiinflamatorios/farmacología , Anticuerpos Monoclonales/farmacología , Anticuerpos Monoclonales/uso terapéutico , Humanos , Lesión Pulmonar/metabolismo , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
DNA methylation patterns may serve as a key in determining cell phenotypes and functions. Adjacent CpG patterns may provide insight into methylation functional mechanisms. Some regions display different DNA methylation patterns between normal and cancer tissues, but the same average methylation level. Here, we developed a method (CellMethy) to infer a region in which all CpGs exhibit concordant methylation (CM) and to quantify the extent of CM in the region. Using simulation data, CellMethy showed high performance in discovering the concordant methylation patterns (AUC = 0.89). CellMethy was then applied to RRBS data including 11 normal tissues and 12 tumors. We found that the extent of CM exhibited wider differentials among tissues than did the average methylation levels from the CM regions, with 45% of CM regions occurring specifically in one tissue and mainly in tumors. Then, we identified CM regions through genome wide bisulfite sequencing (GWBS) data on breast cancer. Approximately 82% of CM regions revealed a significantly different extent of CM between cancer and normal tissues. CellMethy can accurately describe concordantly methylated regions, and the results suggest that CM might also serve as a stable marker of cell sub-populations.
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Islas de CpG/genética , Metilación de ADN/genética , Neoplasias/genética , Línea Celular Tumoral , Células Hep G2 , Humanos , Células Jurkat , Células K562RESUMEN
Interactions between complement anaphylatoxins have been investigated in numerous fields; however, their functions during arterial remodeling following injury have not been studied. The inhibitory effect of complement anaphylatoxin C4a on neointima formation induced by C5a following arterial injury was investigated. Mice were subjected to wire-induced endothelial denudation of the femoral artery and treated with C5a alone or C5a + C4a for two weeks. C4a significantly inhibited C5a-induced neointima formation and the expression of CD68, F4/80, tumor necrosis factor-α (TNFα), interleukin-6 (IL-6) and monocyte chemotactic protein-1 (MCP-1). In vitro, although C4a did not directly inhibit the migration, proliferation or the expression of vascular cell adhesion molecule-1 (VCAM-1) of C5a-induced vascular smooth muscle cells (VSMCs), C5a-pretreated conditioned mediuminduced migration, proliferation and VCAM-1 expression of VSMCs were suppressed when VSMCs were exposed to conditioned medium from C4a-pretreated macrophages. In addition, C5a-induced TNF-α, IL-6 and MCP-1 expression, Ca2+ influx and extracellular signal-regulated kinase (ERK) activation in macrophages were suppressed by C4a. C4a inhibits C5a-induced neointima formation, not by acting directly on VSMCs, but via a macrophage-mediated reaction by inhibiting the Ca2+-dependent ERK pathway in macrophages.
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Complemento C4a/farmacología , Complemento C5a/farmacología , Arteria Femoral/efectos de los fármacos , Neointima/patología , Animales , Calcio/metabolismo , Células Cultivadas , Quimiocinas/metabolismo , Complemento C4a/genética , Complemento C4a/metabolismo , Complemento C5a/genética , Complemento C5a/metabolismo , Citocinas/metabolismo , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Arteria Femoral/lesiones , Arteria Femoral/metabolismo , Macrófagos/citología , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Ratones , Ratones Endogámicos C57BL , Miocitos del Músculo Liso/citología , Miocitos del Músculo Liso/efectos de los fármacos , Miocitos del Músculo Liso/metabolismo , Neointima/etiología , Neointima/metabolismo , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/genética , Proteínas Recombinantes/farmacología , Molécula 1 de Adhesión Celular Vascular/metabolismoAsunto(s)
Inhibidores de la Enzima Convertidora de Angiotensina/farmacología , Fibrilación Atrial/tratamiento farmacológico , Cilazapril/farmacología , Células Endoteliales/efectos de los fármacos , Fibrinólisis/efectos de los fármacos , Animales , Fibrilación Atrial/sangre , Fibrilación Atrial/fisiopatología , Perros , Ecocardiografía , Células Endoteliales/fisiología , Óxido Nítrico/biosíntesis , Inhibidor 1 de Activador Plasminogénico/análisis , Activador de Tejido Plasminógeno/análisisRESUMEN
Coriaria nepalensis, Pteridium aquilinum var. latiuscukum, Imperata cylindrical var. major, and Quercus fabric were used as mulching materials to study their effects on the rhizosphere soil microbial population and enzyme activity and the tree growth in poplar plantation. The results showed that after mulching with test materials, the populations of both bacteria and fungi in rhizosphere soil were more than those of the control. Of the mulching materials, I. cylindrical and Q. fabric had the best effect, with the numbers of bacteria and fungi being 23.56 and 1.43 times higher than the control, respectively. The bacterial and fungal populations in rhizosphere soil increased with increasing mulching amount. When the mulching amount was 7.5 kg m(-2), the numbers of bacteria and fungi in rhizosphere soil were 0.5 and 5.14 times higher than the control, respectively. Under bio-mulching, the bacterial and fungal populations in rhizosphere soil had a similar annual variation trend, which was accorded with the annual fluctuation of soil temperature and got to the maximum in July and the minimum in December. The urease and phosphatase activities in rhizosphere soil also increased with increasing mulching amount. As for the effects of different mulching materials on the enzyme activities, they were in the order of C. nepalensis > P. aquilinum > I. cylindrical > Q. fabric. The annual variation of urease and phosphatase activities in rhizosphere soil was similar to that of bacterial and fungal populations, being the highest in July and the lowest in December. Bio-mulching promoted the tree height, DBH, and biomass of poplar trees significantly.
Asunto(s)
Raíces de Plantas/microbiología , Populus/crecimiento & desarrollo , Microbiología del Suelo , Árboles/crecimiento & desarrollo , Biodiversidad , Ecosistema , Monoéster Fosfórico Hidrolasas/metabolismo , Poaceae/crecimiento & desarrollo , Pteridium/crecimiento & desarrollo , Quercus/crecimiento & desarrollo , Suelo/análisis , Árboles/clasificación , Ureasa/metabolismoRESUMEN
BACKGROUND: Inflammation and oxidant stress have been suggested to be involved in the structural remodeling in atrial fibrillation (AF), and inducible nitric oxide synthase (iNOS) is associated with inflammation and oxidant stress. To study whether iNOS could contribute to atrial remodeling in AF, we investigated the relationship between inflammation, oxidant stress, nitric oxide (NO) and its synthase, and cardiomyocytic apoptosis in the right atrium in human AF. METHODS: Fifteen patients with permanent AF (PmAF) and 17 patients with sinus rhythm (SR), who underwent mitral valve replacement surgery were investigated. Western blotting and immunohistochemical staining were used to detect the expression of endothelial nitric oxide synthase (eNOS), iNOS and 3-nitrotyrosine (3NT; a biological marker of peroxynitrite) in the right atrium. The occurrence of cardiomyocytic apoptosis was determined by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL). Caspase 3 staining for the activated, cleaved protease was also performed. In addition, concentrations of NO(2)(-)/NO(3)(-) (NO(X)) both in plasma and in the right atrium were measured by a NO(X) Detection Kit. Finally, plasma levels of high-sensitivity C-reactive protein (hs-CRP) were routinely measured. RESULTS AND CONCLUSIONS: Levels of hs-CRP were far more enhanced in patients with PmAF compared to the controls. Plasma levels of NO(X) were significantly lower in PmAF patients than SR patients, but the production of NO(X) in the local right atrium increased obviously. Furthermore, iNOS and 3NT expressions increased dramatically in the right atrium in PmAF patients, whereas the expression of eNOS did not change apparently. In addition, when patients were further divided into a higher hs-CRP group (> or =5 mg/L) and a lower hs-CRP group (<5 mg/L) according to the hs-CRP level, significant upregulation of iNOS was found in the higher hs-CRP group. Apoptosis index and caspase 3 staining were also prominently enhanced in PmAF patients compared with SR patients. More importantly, we demonstrated in this study that a higher expression of 3NT was associated with an increased expression of iNOS/eNOS (r=0.74, P<0.05) and an enhanced apoptosis index (r=0.69, P<0.05). In conclusion, the results presented novel evidence that imbalanced expression of iNOS/eNOS could contribute to protein nitration and cardiomyocyte apoptosis in human AF, in which condition inflammation may be an important participant.