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OBJECTIVE AND DESIGN: We aimed to investigate the molecular mechanism underlying formaldehyde (FA)-induced congenital heart disease (CHD) using in vitro and in vivo models. MATERIALS AND SUBJECTS: Neonatal rat heart tissues and H9C2 cells were used for in vitro studies, while FA-exposed new-born rats were used for in vivo studies. TREATMENT: H9C2 cells were exposed to FA concentrations of 0, 50, 100 and 150 µM/mL for 24 h. METHODS: Whole transcriptome gene sequencing identified differentially expressed miRNAs in neonatal rat heart tissues, while Real-time quantitative PCR (RT-qPCR) assessed miR-871-3p and Megf8 expression. RNA pull-down and dual-luciferase reporter assays determined miR-871-3p and Megf8 relationships. Inflammatory cytokine expression was assessed by western blotting. A FA-induced CHD model was used to validate miR-871-3p regulatory effects in vivo. RESULTS: We identified 89 differentially expressed miRNAs, with 28 up-regulated and 61 down-regulated (fold change ≥ 2.0, P < 0.05). Inflammation (interleukin) and signalling pathways were found to control FA-induced cardiac dysplasia. miR-871-3p was upregulated in FA-exposed heart tissues, modulated inflammation, and directly targeted Megf8. In vivo experiments showed miR-871-3p knockdown inhibited FA-induced inflammation and CHD. CONCLUSION: We demonstrated miR-871-3p's role in FA-induced CHD by targeting Megf8, providing potential targets for CHD intervention and improved diagnosis and treatment strategies.
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Formaldehído , Cardiopatías , Proteínas de la Membrana , MicroARNs , Animales , Femenino , Humanos , Lactante , Recién Nacido , Masculino , Ratas , Contaminantes Atmosféricos/metabolismo , Contaminantes Atmosféricos/toxicidad , Modelos Animales de Enfermedad , Formaldehído/metabolismo , Formaldehído/toxicidad , Expresión Génica , Técnicas de Silenciamiento del Gen , Corazón/efectos de los fármacos , Corazón/fisiopatología , Cardiopatías/congénito , Cardiopatías/metabolismo , Cardiopatías/patología , Inflamación/metabolismo , Proteínas de la Membrana/metabolismo , MicroARNs/genética , MicroARNs/metabolismo , Miocitos Cardíacos/efectos de los fármacos , Miocitos Cardíacos/metabolismo , Miocitos Cardíacos/patología , Ratas Sprague-DawleyRESUMEN
AIM: Autophagy plays essential roles in abdominal aortic aneurysm (AAA) development and progression. The objective of this study was to verify the autophagy-related genes (ARGs) underlying AAA empirically and using bioinformatics analysis. METHODS: Two gene expression profile datasets GSE98278 and GSE57691 were downloaded from the Gene Expression Omnibus (GEO) database, and principal component analysis was performed. Following, the R software (version 4.0.0) was employed to analyze potentially differentially expressed genes related with AAA and autophagy. Subsequently, the candidate genes were screened using protein-protein interaction (PPI), gene ontology (GO) enrichment analysis, and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. Finally, quantitative real-time polymerase chain reaction (RT-qPCR) was performed to detect the RNA expression levels of the top five selected abnormal ARGs in clinical samples obtained from the normal and AAA patients. RESULTS: According to the information contained (97 AAA patients and 10 healthy controls) in the two datasets, a total of 44 differentially expressed autophagy-related genes (6 up-regulated genes and 38 down-regulated genes) were screened. GO enrichment analysis of differentially expressed autophagy-related genes (DEARGs) demonstrated that some enrichment items were associated with inflammation, and PPI analysis indicated interaction between these genes. RT-qPCR results presented that the expression levels of IL6, PPARG, SOD1, and MAP1LC3B were in accordance with the bioinformatics prediction results acquired from the mRNA chip. CONCLUSION: Bioinformatics analysis identified 44 potential autophagy-related differentially expressed genes in AAA. Further verification by RT- qPCR presented that IL6, PPARG, SOD1, and MAP1LC3B may affect the development of AAA by regulating autophagy. These findings might help explain the pathogenesis of AAA and be helpful in its diagnosis and treatment.
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Aneurisma de la Aorta Abdominal , Interleucina-6 , Humanos , PPAR gamma , Superóxido Dismutasa-1 , Autofagia/genética , Aneurisma de la Aorta Abdominal/genéticaRESUMEN
Formaldehyde exposure during pregnancy can cause fetal congenital heart disease (CHD). However, the regulatory mechanism remains unclear. Studies on the biology of long non-coding RNAs (lncRNAs) show that lncRNAs can influence cardiac development and disease. However, expression patterns and regulatory mechanisms of action of lncRNAs in formaldehyde-induced CHD remain unclear. We used high-throughput sequencing strategies as a means of identifying lncRNA expression profiles in heart tissues of normal and formaldehyde-exposed newborn rats. Overall, 763 differentially expressed lncRNAs were identified, including 325 and 438 that were respectively up-regulated and down-regulated. GO and KEGG analyses indicated that the Ras and hedgehog signaling pathways may be important regulatory pathways in CHD caused by exposure to formaldehyde. A lncRNA-miRNA-mRNA co-expression network was constructed and a key miRNA, rno-miR-665, was identified. Furthermore, qRT-PCR analysis verified that the novel lncRNAs: MSTRG.27313.2, MSTRG.30629.2, MSTRG.36520.33, MSTRG.91234.1, and MSTRG.91233.9, were upregulated in the formaldehyde-exposed group. These differentially expressed lncRNAs identified during formaldehyde-induced CHD in newborn rats help explain CHD pathogenesis and provide an effective reference for diagnosing and treating CHD.
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MicroARNs , ARN Largo no Codificante , Embarazo , Femenino , Ratas , Animales , ARN Largo no Codificante/genética , Proteínas Hedgehog/genética , MicroARNs/genética , ARN Mensajero/genética , Corazón , Redes Reguladoras de Genes , Perfilación de la Expresión GénicaRESUMEN
Circular RNAs (circRNAs) are a novel type of long non-coding RNAs that can regulate gene expression in heart development and heart disease. However, the expression pattern of circRNAs in congenital heart disease (CHD) induced by formaldehyde exposure is still unknown. We detected circRNAs expression profiles in heart tissue taken from six neonatal rat pups with formaldehyde exposure group and normal group using RNA-sequencing. Results revealed that a total of 54 circRNAs were dysregulated in the formaldehyde exposure group compared to the normal group. Among them, 31 were upregulated and 23 were downregulated (fold change = 2.0, p < 0.0 5). The qRT-qPCR results showed that expressions of 12:628708|632694, 18:77477060|77520779, 5:167486001|167526275 were significantly upregulated, while that of 7:41167312|4116775 and 20:50659751|5068786 were notably downregulated; the expression pattern was consistent with the RNA sequencing data. Bioinformatics analysis shows that the pathogenesis of formaldehyde exposure-induced CHD may involve Hippo-YAP pathwayãNotch signaling pathway and other pathways. A key miRNA (rno-miR-665) was identified by constructing a circRNA-miRNA-mRNA co-expression network. In summary, the study illustrated that circRNAs differentially expressed in fetal heart tissues during formaldehyde exposure has potential biological functions and may be a biomarker or therapeutic target for CHD.
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As a common air pollutant, formaldehyde is widely present in nature, industrial production and consumer products. Endogenous formaldehyde is mainly produced through the oxidative deamination of methylamine catalysed by semicarbazide-sensitive amine oxidase (SSAO) and is ubiquitous in human body fluids, tissues and cells. Vascular endothelial cells and smooth muscle cells are rich in this formaldehyde-producing enzyme and are easily damaged owing to consequent cytotoxicity. Consistent with this, increasing evidence suggests that the cardiovascular system and stages of heart development are also susceptible to the harmful effects of formaldehyde. Exposure to formaldehyde from different sources can induce heart disease such as arrhythmia, myocardial infarction (MI), heart failure (HF) and atherosclerosis (AS). In particular, long-term exposure to high concentrations of formaldehyde in pregnant women is more likely to affect embryonic development and cause heart malformations than long-term exposure to low concentrations of formaldehyde. Specifically, the ability of mouse embryos to effect formaldehyde clearance is far lower than that of the rat embryos, more readily allowing its accumulation. Formaldehyde may also exert toxic effects on heart development by inducing oxidative stress and cardiomyocyte apoptosis. This review focuses on the current progress in understanding the influence and underlying mechanisms of formaldehyde on cardiovascular disease and heart development.
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Enfermedades Cardiovasculares/patología , Desinfectantes/efectos adversos , Formaldehído/efectos adversos , Efectos Tardíos de la Exposición Prenatal/patología , Animales , Enfermedades Cardiovasculares/inducido químicamente , Enfermedades Cardiovasculares/metabolismo , Femenino , Humanos , Embarazo , Efectos Tardíos de la Exposición Prenatal/inducido químicamente , Efectos Tardíos de la Exposición Prenatal/metabolismoRESUMEN
ßII spectrin, the most common isoform of non-erythrocyte spectrin, is a cytoskeleton protein present in all nucleated cells. Interestingly, ßII spectrin is essential for the development of various organs such as nerve, epithelium, inner ear, liver and heart. The functions of ßII spectrin include not only establishing and maintaining the cell structure but also regulating a variety of cellular functions, such as cell apoptosis, cell adhesion, cell spreading and cell cycle regulation. Notably, ßII spectrin dysfunction is associated with embryonic lethality and the DNA damage response. More recently, the detection of altered ßII spectrin expression in tumors indicated that ßII spectrin might be involved in the development and progression of cancer. Its mutations and disorders could result in developmental disabilities and various diseases. The versatile roles of ßII spectrin in disease have been examined in an increasing number of studies; nonetheless, the exact mechanisms of ßII spectrin are still poorly understood. Thus, we summarize the structural features and biological roles of ßII spectrin and discuss its molecular mechanisms and functions in development, homeostasis, regeneration and differentiation. This review highlight the potential effects of ßII spectrin dysfunction in cancer and other diseases, outstanding questions for the future investigation of therapeutic targets. The investigation of the regulatory mechanism of ßII spectrin signal inactivation and recovery may bring hope for future therapy of related diseases.
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Enfermedad/etiología , Neoplasias/metabolismo , Espectrina/metabolismo , Animales , Adhesión Celular , Ciclo Celular , Inestabilidad Genómica , Humanos , Terapia Molecular Dirigida , Neoplasias/tratamiento farmacológico , Espectrina/químicaRESUMEN
Background: Formaldehyde (FA) is ubiquitous in the environment and can be transferred to the fetus through placental circulation, causing miscarriage and congenital heart disease (CHD). Studies have shown that ßII spectrin is necessary for cardiomyocyte survival and differentiation, and its loss leads to heart development defects and cardiomyocyte apoptosis. Additionally, previous studies have demonstrated that miRNA is essential in heart development and remodeling. However, whether miRNA regulates FA-induced CHD and cardiomyocyte apoptosis remains unclear. Methods: Using commercially available rat embryonic cardiomyocytes and a rat model of fetal cardiomyocyte apoptosis. Real-time quantitative PCR (RT-qPCR) and Western blot were performed to examine the level of miR-153-3p, ßII spectrin, caspase 7, cleaved caspase7, Bax, Bcl-2 expression in embryonic cardiomyocytes and a rat model of fetal cardiomyocyte apoptosis. Apoptotic cell populations were evaluated by flow cytometry and Tunel. Luciferase activity assay and RNA pull-down assay were used to detect the interaction between miR-153-3p and ßII spectrin. Masson's trichrome staining detects the degree of tissue fibrosis. Fluorescence in situ hybridization (FISH) and Immunohistochemistry were used to detect the expression of miR-153-3p and ßII spectrin in tissues. Results: Using commercially available rat embryonic cardiomyocytes and a rat model of fetal cardiomyocyte apoptosis, our studies indicate that miR-153-3p plays a regulatory role by directly targeting ßII spectrin to promote cardiomyocyte apoptosis. miR-153-3p mainly regulates cardiomyocyte apoptosis by regulating the expression of caspase7, further elucidating the importance of apoptosis in heart development. Finally, the results with our animal model revealed that targeting the miR-153-3p/ßII spectrin pathway effectively regulated FA-induced damage during heart development. Recovery experiments with miR-153-3p antagomir resulted in the reversal of FA-induced cardiomyocyte apoptosis and fetal cardiac fibrosis. Conclusion: This study investigated the molecular mechanism underpinning the role of ßII spectrin in FA-induced CHD and the associated upstream miRNA pathway. The study findings suggest that miR-153-3p may provide a potential target for the clinical diagnosis and treatment of CHD.
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INTRODUCTION: The performances of contrast-enhanced ultrasound (CEUS) and digital subtraction angiography (DSA) were used to establish an efficient as well as non-invasive clinical technique for the diagnosis of extra-cranial internal carotid artery (ICA) stenosis. MATERIALS AND METHODS: Thirty-six successive patients (11 women and 25 men, mean age: 65.0 ± 9.2, range: 43-78 years) with internal carotid artery (ICA) stenosis were tested by CEUS and DSA. These tests were carried out by means of Hitachi Preirus ultrasound machine for CEUS and Allura Xper FD20 system (Philips Medical Systems, Nederland B.V.) for DSA. 1.2 ml SonoVue (Bracco, Switzerland) was used a s contrast agent. RESULTS: The results clearly indicated that there were no noteworthy variations among the distributions recorded by CEUS as well as DSA for the four tested groups. The percentage of diameter stenosis calculated by CEUS was clearly in accordance with the DSA images. CEUS showed accurate results with good specificity and sensitivity at 50%, 70%, and 100%. Also, CEUS performance was relatively better than DSA in the diagnosis of ICA and suitability of CEA. CONCLUSION: CEUS proved to be a precise non-invasive testing method for the diagnosis of carotid artery stenosis which is more feasible and well-tolerated in patients with various stages of carotid stenosis.
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Angiografía de Substracción Digital/métodos , Estenosis Carotídea/diagnóstico por imagen , Ultrasonografía/métodos , Adulto , Anciano , Estenosis Carotídea/diagnóstico , Medios de Contraste , Femenino , Humanos , Masculino , Persona de Mediana Edad , Valor Predictivo de las Pruebas , Estudios Retrospectivos , Sensibilidad y Especificidad , Índice de Severidad de la EnfermedadRESUMEN
The periosteum plays a vital role in both development and injury healing process of bone. However, few researches have focused on artificial periosteum, which was also limited by the complexity on its construction and biological risks for clinical practice. In order to tackle this issue, inspired by the structural development of periosteum, we put forward a hierarchical micro/nanofibrous bionic periosteum with sustained releasing of VEGF as exogenous vascularized fibrous layer of periosteum to induce endogenous cambium layer in vivo for complete regeneration of periosteal and bone tissue, through collagen self-assembly and micro-sol electrospinning technologies. The VEGF encapsulated in hyaluronan-PLLA core-shell structure was demonstrated to be released in a durable way for angiogenesis in fibrous layer and bone defect area. Meanwhile, the self-assembly of collagen together with electrospun fibers contributed to a hierarchical micro/nanostructure which greatly mimicked the microenvironment of extracellular matrix to provide structural and biochemical cues for cell adhesion, proliferation and differentiation, and lead to the formation of cambium layer which mimicked the in-situ ossification manner as intramembranous ossification. As the motif of this study, the periosteal regeneration was characterized both by osteoblasts and periostin, which represented structural and molecular mechanisms respectively. Furthermore, the periosteal biomaterial proposed here possessed the superior abilities of scar inhibition, angiogenesis, osteogenesis to repair the bone defect in a uniform and rapid manner by inherent periosteal ossific mechanism involved in both intramembranous and endochondral ossification. Thus, the endogenous-exogenous combined bionic periosteum proved to be efficient and versatile in triggering periosteal and bone regeneration and hopefully supply a promising strategy for solving clinical issue.
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Nanofibras , Factor A de Crecimiento Endotelial Vascular , Regeneración Ósea , Preparaciones de Acción Retardada , Sistemas de Liberación de Medicamentos , Osteogénesis , PeriostioRESUMEN
To explore the relationship between risk prediction of diabetes mellitus (DM) and different physical fitness parameters in municipal in-service personnel in Guangxi.This was a cross-sectional study conducted in China from July 2015 to December 2016. We enrolled in-service adults (20-65 year of age) from public institutions. All subjects underwent National Physical Fitness Test (NPFT) and EZSCAN screening.The 5668 subjects were 42.9±12.3 years; 2984 (52.6%) were male; 3998 (70.5%), 1579 (27.9%) and 85 (1.6%) were Han, Zhuang, and other ethnicities, respectively. The multivariable analysis showed that systolic blood pressure (odds ratio [OR]â=â1.013, 95% confidence interval [CI]: 1.003-1.022, Pâ=â.01), Harvard step test index (ORâ=â0.958, 95% CI: 0.941-0.976, Pâ<.001), bend-ahead in sitting (ORâ=â0.945, 95% CI: 0.926-0.963, P <.001), hand grip strength (ORâ=â0.981, 95% CI: 0.966-0.997, Pâ=â.02), vertical jump height (ORâ=â0.969, 95% CI: 0.944-0.996, Pâ=â.02), time of single-leg standing with eyes closed (ORâ=â0.981, 95% CI: 0.968-0.995, Pâ=â.007), choice reaction time (ORâ=â2.103, 95% CI: 1.261-3.507, Pâ=â.004), and body composition minerals (ORâ=â1.649, 95% CI: 1.261-1.813, Pâ<â.001) were independently associated with DM. The resulting equation for the prediction of DM had an area under the receiver operating characteristic curve of 0.808, indicating good predictive ability.NPFT and EZSCAN could help predict the risk of diabetes and give early warnings to undertake preventive actions such as changing diet and performing physical activity.
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Diabetes Mellitus/epidemiología , Aptitud Física , Adulto , Anciano , Presión Sanguínea , Pesos y Medidas Corporales , China , Estudios Transversales , Femenino , Fuerza de la Mano , Humanos , Masculino , Persona de Mediana Edad , Adulto JovenRESUMEN
The apical membrane Cl-/oxalate exchanger SLC26A6 has been demonstrated to play a role in proximal tubule NaCl transport based on studies in microperfused tubules. The present study is directed at characterizing the role of SLC26A6 in NaCl homeostasis in vivo under physiological conditions. Free-flow micropuncture studies revealed that volume and Cl- absorption were similar in surface proximal tubules of wild-type and Slc26a6-/- mice. Moreover, the increments in urine flow rate and sodium excretion following thiazide and furosemide infusion were identical in wild-type and Slc26a6-/- mice, indicating no difference in NaCl delivery out of the proximal tubule. The absence of an effect of deletion of SLC26A6 on NaCl homeostasis was further supported by the absence of lower blood pressure in Slc26a6-/- compared with wild-type mice on normal or low-salt diets. Moreover, raising plasma and urine oxalate by feeding mice a diet enriched in soluble oxalate did not affect mean blood pressure. In contrast to the lack of effect of SLC26A6 deletion on NaCl homeostasis, fractional excretion of oxalate was reduced from 1.6 in wild-type mice to 0.7 in Slc26a6-/- mice. We conclude that, although SLC26A6 is dispensable for renal NaCl homeostasis, it is required for net renal secretion of oxalate.
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Antiportadores/metabolismo , Túbulos Renales Proximales/metabolismo , Ácido Oxálico/orina , Eliminación Renal , Cloruro de Sodio Dietético/orina , Transportadores de Sulfato/metabolismo , Animales , Antiportadores/deficiencia , Antiportadores/genética , Presión Sanguínea , Dieta Hiposódica , Femenino , Genotipo , Homeostasis , Masculino , Ratones de la Cepa 129 , Ratones Noqueados , Fenotipo , Transportadores de Sulfato/deficiencia , Transportadores de Sulfato/genéticaRESUMEN
The study was conducted during the growing seasons of 2013, 2014, and 2015 in the wet meadows on the eastern Qinghai-Tibet plateau (QTP) in the Gansu Gahai Wetland Nature Reserve to determine the dynamics of soil organic carbon (SOC) as affected by vegetation degradation along a moisture gradient and to assess its relationship with other soil properties and biomass yield. Hence, we measured SOC at depths of 0-10, 10-20, and 20-40 cm under the influence of four categories of vegetation degradation (healthy vegetation [HV], slightly degraded [SD], moderately degraded [MD], and heavily degraded [HD]). Our results showed that SOC decreased with increased degree of vegetation degradation. Average SOC content ranged between 36.18 ± 4.06 g/kg in HD and 69.86 ± 21.78 g/kg in HV. Compared with HV, SOC content reduced by 30.49%, 42.22%, and 48.22% in SD, MD, and HD, respectively. SOC significantly correlated positively with soil water content, aboveground biomass, and belowground biomass, but significantly correlated negatively with soil temperature and bulk density (p < 0.05). Highly Significant positive correlations were also found between SOC and total nitrogen (p = 0.0036), total phosphorus (p = 0.0006) and total potassium (p < 0.0001). Our study suggests that severe vegetation and moisture loss led to approximately 50% loss in SOC content in the wet meadows, implying that under climate warming, vegetation and soil moisture loss will dramatically destabilize carbon sink capacities of wetlands. We therefore suggest wetland hydrological management, restoration of vegetation, plant species protection, regulation of grazing activities, and other anthropogenic activities to stabilize carbon sink capacities of wetlands.
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In this study, we investigated the protection effect of Vitamin E (Vit E) on formaldehyde (FA) exposure during pregnancy induced apoptosis of cardiomyocytes, and used an HL-1 cell line to confirmed the findings in vivo.Pregnant mice received different doses of FA (0.5 mg/kg, 1.0 mg/kg, 1.5 mg/kg, 0.1 µg Vit E, or 1.5 mg/kg + 0.1 µg Vit E). TUNEL staining was used to reveal the apoptosis in cardiomyocytes, and SOD, MDA, GSH, Livin, and Caspase-3 in cardiomyocytes were detected by ELISA, RT-PCR, and Western blot. For in vitro study, HL-1 cells were treated with vehicle, 5 µmol/L FA, 25 µmol/L FA, 50 µmol/L FA, 10 mg/L Vit. E, and 50 µmol/L FA+ 10 mg/L Vit E, respectively. CCK-8 assay and flow cytometry were used to evaluate cell vitality and apoptosis. A high dose of FA exposure led to cytotoxicity in both pregnant mice and offspring, as TUNEL staining revealed a significant apoptosis of cardiomyocytes, and the alternation in SOD, GSH, MDA, Livin, and Caspase-3 was found in cardiomyocytes. 0.1 µg Vit. E could reverse high doses of FA exposure induced apoptosis of cardiomyocytes in both pregnant mice and offspring. The in vitro study revealed that FA exposure induced a decrease of cell viability and increased cell apoptosis, as well as oxidative stress in HL-1 cells with alternation in SOD, GSH, MDA, Livin, and Caspase-3.This study revealed a high dose of FA induced oxidative stress and apoptosis of cardiomyocytes in both pregnant mice and offspring, and Vit E supplement during pregnancy reversed the systemic and myocardial toxicity of FA.
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Apoptosis/efectos de los fármacos , Formaldehído/efectos adversos , Miocitos Cardíacos/metabolismo , Estrés Oxidativo/efectos de los fármacos , Preñez , Hipersensibilidad Respiratoria/tratamiento farmacológico , Vitamina E/farmacología , Animales , Animales Recién Nacidos , Antioxidantes/farmacología , Western Blotting , Modelos Animales de Enfermedad , Ensayo de Inmunoadsorción Enzimática , Femenino , Formaldehído/metabolismo , Formaldehído/toxicidad , Etiquetado Corte-Fin in Situ , Ratones , Miocitos Cardíacos/efectos de los fármacos , Miocitos Cardíacos/patología , Embarazo , Complicaciones del Embarazo/inducido químicamente , Complicaciones del Embarazo/metabolismo , Complicaciones del Embarazo/patología , Hipersensibilidad Respiratoria/metabolismo , Hipersensibilidad Respiratoria/patología , Superóxido Dismutasa/metabolismoRESUMEN
Patients with cystic fibrosis have an increased incidence of hyperoxaluria and calcium oxalate nephrolithiasis. Net intestinal absorption of dietary oxalate results from passive paracellular oxalate absorption as modified by oxalate back secretion mediated by the SLC26A6 oxalate transporter. We used mice deficient in the cystic fibrosis transmembrane conductance regulator gene (Cftr) to test the hypothesis that SLC26A6-mediated oxalate secretion is defective in cystic fibrosis. We mounted isolated intestinal tissue from C57BL/6 (wild-type) and Cftr-/- mice in Ussing chambers and measured transcellular secretion of [14C]oxalate. Intestinal tissue isolated from Cftr-/- mice exhibited significantly less transcellular oxalate secretion than intestinal tissue of wild-type mice. However, glucose absorption, another representative intestinal transport process, did not differ in Cftr-/- tissue. Compared with wild-type mice, Cftr-/- mice showed reduced expression of SLC26A6 in duodenum by immunofluorescence and Western blot analysis. Furthermore, coexpression of CFTR stimulated SLC26A6-mediated Cl--oxalate exchange in Xenopus oocytes. In association with the profound defect in intestinal oxalate secretion, Cftr-/- mice had serum and urine oxalate levels 2.5-fold greater than those of wild-type mice. We conclude that defective intestinal oxalate secretion mediated by SLC26A6 may contribute to the hyperoxaluria observed in this mouse model of cystic fibrosis. Future studies are needed to address whether similar mechanisms contribute to the increased risk for calcium oxalate stone formation observed in patients with cystic fibrosis.
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Oxalato de Calcio/metabolismo , Regulador de Conductancia de Transmembrana de Fibrosis Quística/fisiología , Mucosa Intestinal/metabolismo , Animales , Antiportadores/fisiología , Regulador de Conductancia de Transmembrana de Fibrosis Quística/genética , Hiperoxaluria/etiología , Ratones , Ratones Noqueados , Transportadores de SulfatoRESUMEN
OBJECTIVE: To study the role of fine-needle aspiration biopsy and thyroglobulin measurement (FNAB-Tg) in diagnosing cervical lymph node (CLN) metastases in patients with papillary thyroid cancer (PTC) before thyroidectomy. METHODS: Ultrasonography (US)-guided FNAB was performed on 92 patients with PTCs with 105 CLNs before surgery. The wash-out of the FNAB-Tg level was detected. RESULTS: Based on the final pathology, 67 lymph nodes (LNs) were positive for metastasis, and 38 LNs were negative for metastasis. The sensitivity, specificity, accuracy, positive predictive value, and negative predictive value of FNAB-Tg in thyroidectomized patients was 100%, 86.8%, 95.2%, 97.3%, and 95.2%, respectively. CONCLUSIONS: FNAB-Tg is a useful technique for diagnosing CLN metastasis of PTC before thyroidectomy.â©.
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Biomarcadores de Tumor/análisis , Carcinoma Papilar/patología , Carcinoma Papilar/secundario , Biopsia del Ganglio Linfático Centinela/métodos , Tiroglobulina/análisis , Neoplasias de la Tiroides/patología , Adolescente , Adulto , Anciano , Carcinoma Papilar/química , Carcinoma Papilar/cirugía , Femenino , Humanos , Ganglios Linfáticos/química , Metástasis Linfática , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Sensibilidad y Especificidad , Cáncer Papilar Tiroideo , Neoplasias de la Tiroides/química , Neoplasias de la Tiroides/secundario , Neoplasias de la Tiroides/cirugía , Tiroidectomía , Ultrasonografía Intervencional , Adulto JovenRESUMEN
The prevalence of renal stone disease is increasing, although it remains higher in men than in women when matched for age. While still somewhat controversial, several studies have reported an association between renal stone disease and hypertension, but this may be confounded by a shared link with obesity. However, independent of obesity, hyperoxaluria has been shown to be associated with hypertension in stone-formers, and the most common type of renal stone is composed of calcium oxalate. The chloride-oxalate exchanger slc26a6 (also known as CFEX or PAT-1), located in the renal proximal tubule, was originally thought to have an important role in sodium homeostasis and thereby blood pressure control, but it has recently been shown to have a key function in oxalate balance by mediating oxalate secretion in the gut. We have applied two orthogonal analytical platforms (NMR spectroscopy and capillary electrophoresis with UV detection) in parallel to characterize the urinary metabolic signatures related to the loss of the renal chloride-oxalate exchanger in slc26a6 null mice. Clear metabolic differentiation between the urinary profiles of the slc26a6 null and the wild type mice were observed using both methods, with the combination of NMR and CE-UV providing extensive coverage of the urinary metabolome. Key discriminating metabolites included oxalate, m-hydroxyphenylpropionylsulfate (m-HPPS), trimethylamine-N-oxide, glycolate and scyllo-inositol (higher in slc26a6 null mice) and hippurate, taurine, trimethylamine, and citrate (lower in slc26a6 null mice). In addition to the reduced efficiency of anion transport, several of these metabolites (hippurate, m-HPPS, methylamines) reflect alteration in gut microbial cometabolic activities. Gender-related metabotypes were also observed in both wild type and slc26a6 null groups. Urinary metabolites that showed a sex-specific pattern included trimethylamine, trimethylamine-N-oxide, citrate, spermidine, guanidinoacetate, and 2-oxoisocaproate. The gender-dependent metabolic expression of the consequences of slc26a6 deletion might have relevance to the difference in prevalence of renal stone formation in men and women. The different composition of microbial metabolites in the slc26a6 null mice is consistent with the fact that the slc26a6 transporter is found in a range of tissues, including the kidney and intestine, and provides further evidence for the "long reach" of the microbiota in physiological and pathological processes.
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Antiportadores/deficiencia , Metaboloma/fisiología , Metabolómica/métodos , Compuestos Orgánicos/orina , Animales , Antiportadores/genética , Antiportadores/orina , Electroforesis Capilar , Femenino , Masculino , Metaboloma/genética , Ratones , Ratones Noqueados , Resonancia Magnética Nuclear Biomolecular , Compuestos Orgánicos/química , Oxalatos/química , Oxalatos/metabolismo , Fenotipo , Análisis de Componente Principal , Transportadores de SulfatoRESUMEN
Mice deficient for the apical membrane oxalate transporter SLC26A6 develop hyperoxalemia, hyperoxaluria, and calcium oxalate stones due to a defect in intestinal oxalate secretion. However, the nature of the basolateral membrane oxalate transport process that operates in series with SLC26A6 to mediate active oxalate secretion in the intestine remains unknown. Sulfate anion transporter-1 (Sat1 or SLC26A1) is a basolateral membrane anion exchanger that mediates intestinal oxalate transport. Moreover, Sat1-deficient mice also have a phenotype of hyperoxalemia, hyperoxaluria, and calcium oxalate stones. We, therefore, tested the role of Sat1 in mouse duodenum, a tissue with Sat1 expression and SLC26A6-dependent oxalate secretion. Although the active secretory flux of oxalate across mouse duodenum was strongly inhibited (>90%) by addition of the disulfonic stilbene DIDS to the basolateral solution, secretion was unaffected by changes in medium concentrations of sulfate and bicarbonate, key substrates for Sat1-mediated anion exchange. Inhibition of intracellular bicarbonate production by acetazolamide and complete removal of bicarbonate from the buffer also produced no change in oxalate secretion. Finally, active oxalate secretion was not reduced in Sat1-null mice. We conclude that a DIDS-sensitive basolateral transporter is involved in mediating oxalate secretion across mouse duodenum, but Sat1 itself is dispensable for this process.
Asunto(s)
Proteínas de Transporte de Anión/metabolismo , Antiportadores/metabolismo , Duodeno/metabolismo , Oxalatos/metabolismo , Ácido 4,4'-Diisotiocianostilbeno-2,2'-Disulfónico/farmacología , Acetazolamida/farmacología , Animales , Proteínas de Transporte de Anión/deficiencia , Proteínas de Transporte de Anión/genética , Antiportadores/deficiencia , Antiportadores/genética , Transporte Biológico Activo , Ratones , Ratones Noqueados , Transportadores de SulfatoRESUMEN
Mice lacking the oxalate transporter SLC26A6 develop hyperoxalemia, hyperoxaluria, and calcium-oxalate stones as a result of a defect in intestinal oxalate secretion, but what accounts for the absorptive oxalate flux remains unknown. We measured transepithelial absorption of [(14)C]oxalate simultaneously with the flux of [(3)H]mannitol, a marker of the paracellular pathway, across intestine from wild-type and Slc26a6-null mice. We used the anion transport inhibitor DIDS to investigate other members of the SLC26 family that may mediate transcellular oxalate absorption. Absorptive flux of oxalate in duodenum was similar to mannitol, insensitive to DIDS, and nonsaturable, indicating that it is predominantly passive and paracellular. In contrast, in wild-type mice, secretory flux of oxalate in duodenum exceeded that of mannitol, was sensitive to DIDS, and saturable, indicating transcellular secretion of oxalate. In Slc26a6-null mice, secretory flux of oxalate was similar to mannitol, and no net flux of oxalate occurred. Absorptive fluxes of both oxalate and mannitol varied in parallel in different segments of small and large intestine. In epithelial cell lines, modulation of the charge selectivity of the claudin-based pore pathway did not affect oxalate permeability, but knockdown of the tight-junction protein ZO-1 enhanced permeability to oxalate and mannitol in parallel. Moreover, formation of soluble complexes with cations did not affect oxalate absorption. In conclusion, absorptive oxalate flux occurs through the paracellular "leak" pathway, and net absorption of dietary oxalate depends on the relative balance between absorption and SLC26A6-dependent transcellular secretion.