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BACKGROUND: Although the genetic spectrum of human colorectal cancer (CRC) is mainly characterized by APC, KRAS and TP53 mutations, driver genes in tumor initiation have not been conclusively demonstrated. In this study, we aimed to identify novel markers for CRC. METHODS: We performed exome analysis of sporadic colorectal cancer (sCRC) coding regions to screen loss of function (LoF) mutation genes, and carried out systems-level approaches to confirm top rank gene in this study. RESULTS: We identified loss of BMP5 is an early event in CRC. Deep sequencing identified BMP5 was mutated in 7.7% (8/104) of sCRC samples, with 37.5% truncating mutation frequency. Notably, BMP5 negative expression and its prognostic value is uniquely significant in sCRC but not in other tumor types. Furthermore, BMP5 expression was positively correlated with E-cadherin in CRC patients and its dysregulation play a vital role in epithelial-mesenchymal transition (EMT), thus triggering tumor initiation and development. RNA sequencing identified, independent of BMP/Smads pathway, BMP5 signaled though Jak-Stat pathways to inhibit the activation of oncogene EPSTI1. CONCLUSIONS: Our result support a novel concept that the importance of BMP5 in sCRC. The tumor suppressor role of BMP5 highlights its crucial role in CRC initiation and development.
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Proteína Morfogenética Óssea 5/genética , Neoplasias Colorretais/genética , Proteínas Supressoras de Tumor/genética , Linhagem Celular Tumoral , Neoplasias Colorretais/metabolismo , Transição Epitelial-Mesenquimal/genética , Perfilação da Expressão Gênica/métodos , Células HCT116 , Células HT29 , Humanos , Mutação/genética , Transdução de Sinais , Proteínas Smad/genética , TranscriptomaRESUMO
OBJECTIVES: Endothelial dysfunction is involved in the development of the acute coronary syndrome (ACS). Plasma microparticles(MPs) from other diseases have been demonstrated to initiate coagulation and endothelial dysfunction.However, whether MPs from ACS patients impair vasodilatation and endothelial function remains unclear. METHODS: Patients(n = 62) with ACS and healthy controls (n = 30) were recruited for MP isolation. Rat thoracic aortas were incubated with MPs from ACS patients or healthy controls to determine the effects of MPs on endothelial-dependent vasodilatation,the phosphorylation of Akt and endothelial nitric oxide synthase (eNOS), the interaction of eNOS with heat shock protein 90 (Hsp90), and nitric oxide (NO) and superoxide anion(O 2 ) production. The origin of MPs was assessed by flow cytometry. RESULTS: MP concentrations were increased in patients with ACS compared with healthy controls. They were positively correlated with the degree of coronary artery stenosis. MPs from ACS patients impair endothelial-dependent vasodilatation, decrease both Akt and eNOS phosphorylation,decrease the interaction between eNOS and Hsp90,and decrease NO production but increase O 2 generation in rat thoracic aortas. Endothelial-derived MPs and platelet-derived MPs made up nearly 75% of MPs. CONCLUSIONS: Our data indicate that MPs from ACS patients negatively affect endothelial-dependent vasodilatation via Akt/eNOS-Hsp90 pathways.
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Síndrome Coronariana Aguda/metabolismo , Micropartículas Derivadas de Células/metabolismo , Endotélio Vascular/fisiopatologia , Proteínas de Choque Térmico HSP90/metabolismo , Óxido Nítrico Sintase Tipo III/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Vasodilatação/fisiologia , Síndrome Coronariana Aguda/cirurgia , Adulto , Idoso , Animais , Estudos de Casos e Controles , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Óxido Nítrico/metabolismo , Fosforilação , Ratos , Transdução de Sinais , Superóxidos/metabolismoRESUMO
BACKGROUND: The association between dietary cholesterol consumption and dyslipidemia is still in controversy. The study aims to evaluate whether dietary cholesterol intake associated with dyslipidemia and its components in Chinese health examinees. METHODS: A large-scale cross-sectional study was conducted among health examinees of in Shaanxi province. Totally of 8358 participants (3677 male and 4681 female) were included. Dietary cholesterol intake was assessed by validated food frequency questionnaire. Multivariable regression and restricted cubic spline models were used to capture the linear and non-linear association between dietary cholesterol and dyslipidemia. RESULTS: A total of 2429 (29.1%) subjects were newly diagnosed of dyslipidemia, the prevalence was 29.2% in male and 27.7% in female. Mean intake of dietary cholesterol was 213.7 mg/day. After adjusting for all potential confounders including demographics information and lifestyles, higher cholesterol consumption was related to lower risk of dyslipidemia, the ORs (95% CIs) across Q2 to Q4 group were 0.87 (0.60-1.26), 0.80 (0.55-1.18) and 0.61 (0.41-0.91) in female. With further controlling for nutrients principal components, a null association was observed between dietary cholesterol and dyslipidemia and serum lipids, regardless of gender. Results of restricted cubic splines showed that the risk of dyslipidemia decreased slowly until around 300 mg/day in men and 200 mg/day in women, although the non-linear association was not significant. CONCLUSIONS: The study suggested that dietary cholesterol consumption was not associated with dyslipidemia or serum lipids in Chinese health examinees, although a decreased risk was observed before the threshold points.
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Colesterol na Dieta , Dislipidemias , Povo Asiático , China/epidemiologia , Colesterol na Dieta/efeitos adversos , Estudos Transversais , Dislipidemias/epidemiologia , Dislipidemias/etiologia , Feminino , Humanos , MasculinoRESUMO
BACKGROUND: C1q/tumor necrosis factor-related protein 3 (CTRP3) has been reported to be a crucial regulator in myocardial infarction. Nevertheless, the potential molecular mechanism of CTRP3 in ischemia/reperfusion (I/R) injury remains largely unclear. METHODS: The cell model of myocardial I/R injury was established by oxygen-glucose deprivation/reoxygenation (OGD/R) of rat cardiomyocyte H9C2. Expression of CTRP3 and lysosomal-associated membrane protein 1 (LAMP1) was detected in H9C2 cells treated with oxygen-glucose deprivation/reoxygenation (OGD/R). H9C2 cells were transfected with overexpression plasmids of CTRP3 (pcDNA-CTRP3) and LAMP1 (pcDNA-LAMP1), or CTRP3 small interfering RNA (si-CTRP3) or/and pcDNA-LAMP1, and cell proliferation, apoptosis and oxidative stress were testified. Co-IP assay was performed to validate the relationship among CTRP3, LAMP1 and JIP2. The role of CTRP3 and LAMP1 in JIP2/JNK pathway was evaluated with Western blot assay. Furthermore, in vivo myocardial I/R injury model was constructed to investigate the effect of CTRP3. RESULTS: Overexpression of CTRP3 and LAMP1 both significantly promoted cell proliferation, inhibited apoptosis and the production of reactive oxygen species (ROS), malondialdehyde (MAD) and cardiac troponin (cTn-I), while silencing CTRP3 exerted the opposite effects, and LAMP1 overexpression reversed the effect of silencing CTRP3 on the aspects above. CTRP3 interacted with LAMP1, and both CTRP3 and LAMP1 bound with JIP2. SP600125 (JNK inhibitor) could restore the effects of CTRP3 or LAMP1 overexpression on the expression of JIP2 and phosphorylated-JNK (p-JNK), proliferation and apoptosis. Moreover, overexpression of CTRP3 improved cardiac I/R injury in vivo. CONCLUSION: CTRP3 alleviates cardiac I/R injury by elevating LAMP1 and activating JIP2/JNK signaling pathway, which may serve as a potential therapeutic target for I/R injury.
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Infarto do Miocárdio , Traumatismo por Reperfusão Miocárdica , Animais , Apoptose , Glucose/metabolismo , Proteína 1 de Membrana Associada ao Lisossomo/metabolismo , Sistema de Sinalização das MAP Quinases , Infarto do Miocárdio/metabolismo , Traumatismo por Reperfusão Miocárdica/metabolismo , Miócitos Cardíacos/metabolismo , Oxigênio/metabolismo , Ratos , Transdução de SinaisRESUMO
Myocardial ischemia reperfusion (I/R) injury is an important complication of myocardial infarction reperfusion therapy, and no effective treatment has been identified. Based on preexisting evidence, C1q/tumor necrosis factor-related protein 3 (CTRP3) has been reported to be closely associated with myocardial dysfunction. In this study, we found that CTRP3 was downregulated in acute coronary syndrome (ACS) patients and myocardial I/R mice. Silence of CTRP3 aggravated cardiac systolic function due to I/R of mice, while CTRP3 overexpression ameliorated cardiac function. Moreover, overexpression of CTRP3 improved I/R inhibitory effects on the levels of creatinine phosphokinase (CPK), lactate dehydrogenase (LDH) and cardiac troponin-I (cTn-I), myocardial infarction area, the intensity of the 3-nitrotyrosine (3-NT), apoptosis and protein levels of LAMP1, JNK-Interacting Protein-2 (JIP-2) and JNK, while these effects could be exacerbated by downregulation of CTRP3. Co-IP experiments could identify physical interactions between CTRP3 and lysosomal-associated membrane protein 1 (LAMP1) and Numb and JIP2. LAMP1 silence aggravated the inhibition effects of I/R on JIP2 and JNK protein expression, CPK, LDH and cTn-I levels and caspase-3 activity, while overexpression of LAMP1 recovered these inhibition effects of I/R. JNK inhibitor (SP600125) could reverse the inhibitory effects of CTRP3 overexpression on CPK, LDH, cTn-I, myocardial infarction, strong positive staining for 3-NT and apoptosis. These findings demonstrated that CTRP3 protected against injury caused by myocardial I/R through activating LAMP1/JIP2/JNK pathway to attenuate myocardial injury, improve left ventricular function, decrease myocardial infarction, and reduce myocardial apoptosis.
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Infarto do Miocárdio , Isquemia Miocárdica , Traumatismo por Reperfusão Miocárdica , Adipocinas , Animais , Apoptose , Humanos , Proteína 1 de Membrana Associada ao Lisossomo , Proteínas de Membrana Lisossomal/metabolismo , Sistema de Sinalização das MAP Quinases/fisiologia , Camundongos , Infarto do Miocárdio/patologia , Traumatismo por Reperfusão Miocárdica/metabolismo , Miocárdio/patologia , Fatores de Transcrição/metabolismo , Fatores de Necrose TumoralRESUMO
An innovative, ternary nanocomposite composed of overoxidized poly(3,4-ethylenedioxythiophene) (OPEDOT), gold nanoparticles (AuNPs), and electrochemically reduced graphene oxide (ERGO) was prepared on a glassy carbon electrode (GCE) (OPEDOT-AuNPs-ERGO/GCE) through homogeneous chemical reactions and heterogeneous electrochemical methods. The morphology, composition, and structure of this nanocomposite were characterized by transmission electron microscopy, scanning electron microscopy, X-ray diffraction, and X-ray photoelectron spectroscopy. The electrochemical properties of the OPEDOT-AuNPs-ERGO/GCE were investigated by cyclic voltammetry using potassium ferricyanide and hexaammineruthenium(III) chloride redox probe systems. This modified electrode shows excellent electro-catalytic activity for dopamine (DA) and uric acid (UA) under physiological pH conditions, but inhibits the oxidation of ascorbic acid (AA). Linear voltammetric responses were obtained when DA concentrations of approximately 4.0-100 µM and UA concentrations of approximately 20-100 µM were used. The detection limits (S/N=3) for DA and UA were 1.0 and 5.0 µM, respectively, under physiological conditions and in the presence of 1.0 mM of AA. This developed method was applied to the simultaneous detection of DA and UA in human urine, where satisfactory recoveries from 96.7% to 105.0% were observed. This work demonstrates that the developed OPEDOT-AuNPs-ERGO ternary nanocomposite, with its excellent ion-selectivity and electro-catalytic activity, is a promising candidate for the simultaneous detection of DA and UA in the presence of AA in physiological and pathological studies.
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OBJECTIVE: Cardiovascular disease is a major cause of death. This study evaluated the relationship between serum cystatin-c and coronary lesion severity in coronary artery disease (CAD) patients with a normal glomerular filtration rate. METHODS: Nine hundred and fifty-nine patients were retrospectively included and divided into non-CAD and CAD groups according to coronary angiography results. CAD patients were classified into three groups by Gensini score tertiles. Multivariable logistic regression was used to study the relationship between serum cystatin-c and coronary lesion severity. RESULTS: Serum cystatin-c levels were significantly higher in CAD patients than in non-CAD patients. Correlation analysis revealed significant correlations between serum cystatin-c levels with the Gensini score and the number of diseased vessels. The area under the receiver operating characteristic curve of serum cystatin-c was 0.544 and 0.555 for predicting a high Gensini score and three-vessel disease, respectively. Multivariate stepwise regression analysis demonstrated that the serum cystatin-c level was an independent predictor of a high Gensini score [odds ratio (OR) = 2.177, 95% confidence interval (CI) 1.140-3.930] and three-vessel disease (OR = 1.845, 95% CI 0.994-3.424) after adjusting for the conventional CAD risk factors. CONCLUSIONS: Serum cystatin-c was elevated in CAD patients and may be an independent predictor of CAD severity.
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Doença da Artéria Coronariana , Biomarcadores , Angiografia Coronária , Doença da Artéria Coronariana/diagnóstico por imagem , Taxa de Filtração Glomerular , Humanos , Estudos Retrospectivos , Fatores de Risco , Índice de Gravidade de DoençaRESUMO
Small-diameter artificial vascular grafts modified with layer-by-layer (LBL) coating show promise in reducing the failure caused by thrombosis and inflammation, but undesirable stability and bioactivity issues of the coating and payload usually limits their long-term efficacy. Herein, inspired by catechol/gallol surface chemistry, a sandwiched layer-by-layer coating constructed by polyethyleneimine (PEI) and heparin with the embedding of epigallocatechin gallate (EGCG)-dexamethasone combination was used to modify the electrospun polycaprolactone (PCL) vascular grafts. Polyphenol embedding endowed the coating with abundant intermolecular interactions between each coating components, mainly contributed by the π-π stacking, weak intermolecular cross-linking and enriched hydrogen bonding, which further enhanced the coating stability and also supported the sustained release of the payloads, like polyelectrolytes and drugs. Compared with the conventional LBL coating, the loading amounts of heparin and dexamethasone in the EGCG embedded LBL coatings doubled and the drug release could be significantly prolonged without serious initial burst. The in vitro and ex vivo assays indicated that the modified PCL vascular grafts would address impressive prolonged anti-platelet adhesion/activation and anti-fibrinogen denaturation ability. Meanwhile, the dexamethasone loading entrusted the sandwiched LBL coating with mild tissue response, in terms of inhibiting the macrophage activation. These results strongly demonstrated that the sandwiched LBL coating with EGCG embedding was an effective method to improve the patency rates of PCL small artificial vascular grafts, which could also be extended to other blood-contacting materials.
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Catequina , Poliésteres , Catequina/análogos & derivados , Materiais Revestidos Biocompatíveis , HeparinaRESUMO
Nitric oxide generation is considered to be a key factor to mimic endothelial function in terms of anti-coagulation and anti-hyperplasia. Herein, ebselen which could play the similar role as glutathion peroxidase-like was loaded into micelles and was further assembled into a layer-by-layer coating. The ability of nitric oxide generation and corresponding biological effect were investigated. Endothelial-mimetic surface has now attracted huge attention in blood-contacting materials, due to its inherent ability of secreting nitric oxide. Among those categories, nitric oxide generation surface is considered to be safe and tunable in the modification of vascular biomedical devices. How to adsorb or immobilize glutathion peroxidase-like catalyst and maintain sustained/safe nitric oxide generation is full of interest. This study aimed at developing a functional coating constructed via layer-by-layer assembly to introduce the catalyst into the coating by pre-loading ebselen in micelles. We firstly introduced phenylboronic acid moiety into the micelle molecule backbone and grafted catechol moiety to chitosan backbone. Then, chitosan, micelles (containing ebselen) and heparin were adopted as polyelectrolytes and then alternatively assembled onto the substrate via layer-by-layer protocol. The catechol was conjugated to the amine groups of chitosan by Schiff base reaction to synthesize chitosan-catechol. The hydrophobic cholesterol was conjugated to the one end of the hydrophilic hyaluronic acid, and the hydroxymethylphenylboronic acid was conjugated to the other end via the esterification of carboxyl (-COOH) and hydroxyl (-OH). The modified hyaluronic acid could spontaneously form micelles in aqueous solution. Ebselen was the loaded into the as-prepared micelles. Chitosan-catechol, heparin, and micelles were alternatively assembled onto the substrate layer by layer to form a micelle-embedded coating. The micelle-embedded coating with ebselen was successfully obtained and the nitric oxide generation ability was in a safe level which was close to healthy endothelial cells. The coating could effectively inhibit platelet adhesion and smooth muscle cell proliferation. The use of ebselen preloaded into micelles could provide a sustained release of catalyst for in situ nitric oxide generation. Besides, this method could also be used to load diverse drugs and regulate desired properties. The study was approved by the Institutional Review Board of the West China Hospital in Sichuan University on March 3, 2018, with approval No. K2018044.
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Azóis/química , Micelas , Óxido Nítrico/metabolismo , Compostos Organosselênicos/química , Animais , Antioxidantes/farmacologia , Azóis/farmacologia , Plaquetas/citologia , Plaquetas/metabolismo , Catecóis/química , Adesão Celular/efeitos dos fármacos , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Quitosana/química , Materiais Revestidos Biocompatíveis/química , Células Endoteliais/citologia , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/metabolismo , Heparina/química , Humanos , Ácido Hialurônico/química , Isoindóis , Miócitos de Músculo Liso/citologia , Miócitos de Músculo Liso/metabolismo , Compostos Organosselênicos/farmacologia , CoelhosRESUMO
OBJECTIVE: To investigate the relationship between human beta2-Adrenergic Receptor (ADRB2) gene C659G polymorphism and essential hypertension in Xinjiang Kazakans. METHODS: Polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) methods were used to detect the C659G polymorphism of ADRB2 gene in 435 Kazakans including 273 hypertensives (EH) and 162 normotensives (NT) and genotype frequencies between EH and NT were analyzed. RESULTS: The genotype frequencies (CC, CG, GG) of the C659G allele were 85.75%, 13.79%, 0.64% respectively and the C659 and G659 allele frequencies were 7.36%, 92.64% in this cohort. The ADRB2 genotype distribution and the allele frequencies of C659 and G659 were significantly higher in EH than those in NT (all P < 0.05). The G allele is a risk factor contributed to hypertension (OR 12.37). After adjustment for age and BMI, the systolic and diastolic blood pressure levels were significant higher in CG + GG genotype group compared with CC genotype group (P < 0.05). CONCLUSION: There was significant association between the C659G polymorphism of ADRB2 gene and essential hypertension in Xinjiang Kazakans suggesting a role of ADRB2 gene C659G polymorphism in the development of hypertension in Xinjiang Kazakans.
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Hipertensão , Polimorfismo Genético , Estudos de Casos e Controles , Frequência do Gene , Humanos , Receptores AdrenérgicosRESUMO
Sporadic colorectal cancer (sCRC) is one of the most commonly diagnosed cancers worldwide, but few genetic markers have been identified and used for its early detection. MicroRNAs are diverse cellular regulators in cancer pathogenesis that bind to the 3'-untranslated region (3'-UTR) of their target mRNAs, and variants within the miRNA target sites on sCRC-related genes may influence its pathogenesis. To investigate this possibility, we used a bioinformatical method to screen SNPs for putative changes in miRNA recognition sites within the 3'-UTR of sCRC-related genes. The rs11466537 single nucleotide polymorphism was predicted to modify the regulation of hsa-miR-1193 on the Transforming Growth Factor ß Receptor II (TGFBR2) gene. Additionally, luciferase reporter assays indicated that hsa-miR-1193 bound the T allele more strongly than the A allele of rs11466537 (with A being the less frequent variant), and real time-polymerase chain reaction and western blot analysis showed that TGFBR2 is significantly repressed by hsa-miR-1193. Furthermore, overexpression of hsa-miR-1193 promoted HT-29 cell proliferation, while the loss of hsa-miR-1193 inhibited the process. Finally, the rs11466537 genotyping result revealed that the frequency of A allele carriers was 1.5% in the control blood samples, but 0 in the sCRC patients' normal colon tissue samples. Our results demonstrated that hsa-miR-1193 may be involved in sCRC tumourigenesis at least in part by suppression of TGFBR2, and the A allele of rs11466537 disturbed the regulation of hsa-miR-1193 on TGFBR2.