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1.
Med Sci Monit ; 26: e921233, 2020 Feb 07.
Artigo em Inglês | MEDLINE | ID: mdl-32032347

RESUMO

BACKGROUND Osteosarcoma is a common malignant tumor of musculoskeletal stromal cells. Osteosarcoma clinical behavior depends mostly on the histologic grade, the site of primary tumor, the response to chemotherapy, and the presence of pulmonary metastases. The aim of this study was to knockout SHOX CNE9/10 in U2OS osteosarcoma cells and to analyze the effects on cell growth and apoptosis. MATERIAL AND METHODS U2OS cells with CNE9 knockout and U2OS cells with CNE10 knockout were established via the CRISPR/Cas9 system. Sanger sequencing was used to detect the success of the knockdown experiment. Western blotting and quantitative polymerase chain reaction were used to detect the expression levels of short stature homeobox-containing gene (SHOX) protein and messenger RNA (mRNA) after knockdown of CNE9 and CNE10. The cell viability and apoptotic rate were detected by the Cell Counting Kit-8 method and by flow cytometry. RESULTS The Sanger sequencing results showed that the knockdown experiment was successful. The levels of SHOX mRNA and protein were significantly reduced after knocking down CNE9 and CNE10. Knockdown of CNE9 and CNE10 significantly increased the growth and inhibited the apoptosis of U2OS osteosarcoma cells. CNE9/CNE10 knockdown U2OS cells were successfully constructed. CONCLUSIONS Knockdown of CNE9 and CNE10 promoted U2OS cell growth and inhibited apoptosis by decreasing SHOX expression. This CNE9/CNE10 knockout U2OS cell model could provide a bridge for the research on SHOX and CNEs in osteosarcoma.

2.
J Food Sci ; 83(5): 1215-1220, 2018 May.
Artigo em Inglês | MEDLINE | ID: mdl-29660845

RESUMO

In this study, gellan gums with different acyl contents were prepared, and their effects on blueberry cloudy juices were compared. The rheological properties, stability coefficients, sedimentations, color parameters and particle size distributions of blueberry cloudy juices with 0.035% (w/w) of gellan gum were measured. As the acyl content increased, the viscosity increased, and the sedimentation and stability coefficient values decreased. Gellan gums with higher acyl contents provided better beverage stabilizing capabilities through stricter enforcement of the molecular associations. Overall, this study provides valuable information for enhancing the gelation and stabilization of blueberry cloudy juices, confirms the superiority of high acyl gellan gums for inhibiting the color fading of anthocyanins, and further guides the development of novel product concepts. PRACTICAL APPLICATION: Cloudy juices made from blueberries have many benefits. However, the particles in cloudy juices will flocculate during storage, resulting in an undesirable precipitate. In our work, gellan gums with different acyl contents were prepared and applied to blueberry juice to prevent aggregation. The results provide valuable information for enhancing the stabilization of blueberry juices, confirm the superiority of high acyl gellan for inhibiting color fading, and further guide the development of novel product concepts.


Assuntos
Mirtilos Azuis (Planta) , Sucos de Frutas e Vegetais/análise , Polissacarídeos Bacterianos/química , Reologia , Antocianinas/análise , Cor , Aditivos Alimentares/análise , Frutas , Géis/química , Modelos Teóricos , Tamanho da Partícula , Paladar
3.
Insect Sci ; 25(6): 946-958, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28569426

RESUMO

The diamondback moth, Plutella xylostella (L.), uses sulfatases (SULF) to counteract the glucosinolate-myrosinase defensive system that cruciferous plants have evolved to deter insect feeding. Sulfatase activity is regulated by post-translational modification of a cysteine residue by sulfatase modifying factor 1 (SUMF1). We identified 12 SULF genes (PxylSulfs) and two SUMF1 genes (PxylSumf1s) in the P. xylostella genome. Phylogenetic analysis of SULFs and SUMFs from P. xylostella, Bombyx mori, Manduca sexta, Heliconius melpomene, Danaus plexippus, Drosophila melanogaster, Tetranychus urticae and Homo sapiens showed that the SULFs were clustered into five groups, and the SUMFs could be divided into two groups. Profiling of the expression of PxylSulfs and PxylSumfs by RNA-seq and by quantitative real-time polymerase chain reaction showed that two glucosinolate sulfatase genes (GSS), PxylSulf2 and PxylSulf3, were primarily expressed in the midgut of 3rd- and 4th-instar larvae. Moreover, expression of sulfatases PxylSulf2, PxylSulf3 and PxylSulf4 were correlated with expression of the sulfatases modifying factor PxylSumf1a. The findings from this study provide new insights into the structure and expression of SUMF1 and PxylSulf genes that are considered to be key factors for the evolutionary success of P. xylostella as a specialist herbivore of cruciferous plants.


Assuntos
Regulação Enzimológica da Expressão Gênica , Proteínas de Insetos/química , Proteínas de Insetos/metabolismo , Mariposas/enzimologia , Sulfatases/química , Sulfatases/metabolismo , Sequência de Aminoácidos , Animais , Sequência Conservada , Proteínas de Insetos/genética , Mariposas/metabolismo , Especificidade de Órgãos , Filogenia , Domínios Proteicos , Sulfatases/genética
4.
J Affect Disord ; 195: 75-81, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-26874244

RESUMO

BACKGROUND: Major depressive disorder (MDD) is a serious debilitating psychiatric disorder. However, the molecular mechanisms of MDD remain largely unknown, and no objective laboratory-based tests are available to diagnose this disorder. METHODS: A gas chromatography-mass spectrometry (GC-MS) based metabolomic approach was used to compare peripheral blood mononuclear cells (PBMC) metabolic profiling of 50 first onset drug-naïve MDD subjects and 50 healthy controls (training samples), to identify potential metabolite biomarkers for MDD. An independent sample cohort including 58 MDD patients, 40 schizophrenia (SCZ) patients and 56 healthy controls (test samples) was used to validate diagnostic generalizability and specificity of identified biomarkers. RESULTS: 17 PBMC metabolites responsible for discriminating MDD group from healthy control group were identified. These metabolites were mainly involved in disturbances of energy and neurotransmitter metabolism. This PBMC metabolite signature could effectively discriminate MDD subjects from the healthy controls with an AUC of 0.926 in training samples and 0.870 in test samples. Moreover, this metabolite signature enabled distinguishing MDD subjects from schizophrenia subjects with an AUC of 0.899. LIMITATIONS: This study was limited by potential confounding effects of different drug treatments in some MDD and schizophrenia subjects, and lack of animal studies to further validate the identified metabolite pathways in MDD. CONCLUSION: These findings suggest that early disturbances of PBMC energy and neurotransmitter metabolism may be associated with the onset of MDD. This PBMC metabolite signature may facilitate development of a laboratory-based diagnostic test for MDD.


Assuntos
Biomarcadores/sangue , Transtorno Depressivo Maior/sangue , Transtorno Depressivo Maior/diagnóstico , Metabolômica/métodos , Adulto , Animais , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Leucócitos Mononucleares/metabolismo , Masculino , Neurotransmissores/metabolismo , Esquizofrenia/sangue , Esquizofrenia/diagnóstico , Sensibilidade e Especificidade , alfa-Tocoferol/metabolismo
5.
J Transl Med ; 13: 226, 2015 Jul 14.
Artigo em Inglês | MEDLINE | ID: mdl-26169624

RESUMO

BACKGROUND: Schizophrenia is a widespread and debilitating mental disorder. However, the underlying molecular mechanism of schizophrenia remains largely unknown and no objective laboratory tests are available to diagnose this disorder. The aim of the present study was to characterize the alternations of glucose metabolites and identify potential diagnostic biomarkers for schizophrenia. METHODS: Gas chromatography/mass spectrometry based targeted metabolomic method was used to quantify the levels of 13 glucose metabolites in peripheral blood mononuclear cells (PBMCs) derived from healthy controls, schizophrenia and major depression subjects (n = 55 for each group). RESULTS: The majority (84.6%) of glucose metabolites were significantly disturbed in schizophrenia subjects, while only two (15.4%) glucose metabolites were differently expressed in depression subjects relative to healthy controls in both training set (n = 35/group) and test set (n = 20/group). Antipsychotics had only a subtle effect on glucose metabolism pathway. Moreover, ribose 5-phosphate in PBMCs showed a high diagnostic performance for first-episode drug-naïve schizophrenia subjects. CONCLUSION: These findings suggested disturbance of glucose metabolism may be implicated in onset of schizophrenia and could aid in development of diagnostic tool for this disorder.


Assuntos
Glucose/metabolismo , Leucócitos Mononucleares/metabolismo , Metabolômica/métodos , Esquizofrenia/metabolismo , Adulto , Biomarcadores/metabolismo , Estudos de Casos e Controles , Demografia , Transtorno Depressivo Maior/metabolismo , Feminino , Humanos , Masculino , Metaboloma
6.
Sci Rep ; 4: 5855, 2014 Jul 28.
Artigo em Inglês | MEDLINE | ID: mdl-25068480

RESUMO

Bipolar disorder (BD) is a debilitating mental disorder that cannot be diagnosed by objective laboratory-based modalities. Our previous studies have independently used nuclear magnetic resonance (NMR)-based and gas chromatography-mass spectrometry (GC-MS)-based metabonomic methods to characterize the urinary metabolic profiles of BD subjects and healthy controls (HC). However, the combined application of NMR spectroscopy and GC-MS may identify a more comprehensive metabolite panel than any single metabonomic platform alone. Therefore, here we applied a dual platform (NMR spectroscopy and GC-MS) that generated a panel of five metabolite biomarkers for BD-four GC-MS-derived metabolites and one NMR-derived metabolite. This composite biomarker panel could effectively discriminate BD subjects from HC, achieving an area under receiver operating characteristic curve (AUC) values of 0.974 in a training set and 0.964 in a test set. Moreover, the diagnostic performance of this panel was significantly superior to the previous single platform-derived metabolite panels. Thus, the urinary biomarker panel identified here shows promise as an effective diagnostic tool for BD. These findings also demonstrate the complementary nature of NMR spectroscopy and GC-MS for metabonomic analysis, suggesting that the combination of NMR spectroscopy and GC-MS can identify a more comprehensive metabolite panel than applying each platform in isolation.


Assuntos
Transtorno Bipolar/diagnóstico , Transtorno Bipolar/urina , Metaboloma , Metabolômica/estatística & dados numéricos , Adulto , Área Sob a Curva , Biomarcadores/urina , Transtorno Bipolar/fisiopatologia , Estudos de Casos e Controles , Análise Discriminante , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Imagem por Ressonância Magnética , Masculino , Metabolômica/métodos , Curva ROC
7.
Artigo em Inglês | MEDLINE | ID: mdl-24657409

RESUMO

A high-performance liquid chromatographic-tandem mass spectrometric (HPLC-MS/MS) method was developed and validated to determine sitafloxacin in human plasma with dextrorphan as internal standard. Chromatographic separation was performed on a ZORBAX SB-C18 column (3.5µm, 2.1mm×100mm) with the mobile phase of methanol/water (containing 0.1% formic acid) (46:54, v/v) at a flow rate of 0.2mL/min. Quantification was performed using multiple-reaction monitoring of the transitions at m/z 410.2 → 392.2 for sitafloxacin and m/z 258.1 → 157.1 for dextrorphan, respectively. The calibration curve was linear over the range of 5-2500ng/mL with the lower limit of quantification of 5ng/mL for sitafloxacin. The intra- and inter-day precisions were less than 8.3% and the deviations of assay accuracies were within ±4.1%. Sitafloxacin was sufficiently stable under all relevant analytical conditions. This method was successfully applied to the pharmacokinetic study of sitafloxacin in healthy Chinese volunteers.


Assuntos
Cromatografia Líquida/métodos , Fluoroquinolonas/sangue , Espectrometria de Massas em Tandem/métodos , Adulto , Estabilidade de Medicamentos , Feminino , Fluoroquinolonas/química , Fluoroquinolonas/farmacocinética , Humanos , Limite de Detecção , Modelos Lineares , Masculino , Reprodutibilidade dos Testes , Adulto Jovem
8.
Mol Biosyst ; 10(4): 813-9, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24457555

RESUMO

Bipolar disorder (BD) is a common and debilitating mental disorder. However, there are no biomarkers available to aid in the diagnosis of this disorder. Here, we used a gas chromatography-mass spectrometry (GC-MS) based metabonomic method to characterize the urinary metabolic profiling of BD subjects and healthy controls to identify and validate urinary metabolite biomarkers for BD. Multivariate statistical analysis was used to visualize group discrimination and identify differentially expressed urinary metabolites in BD subjects relative to the healthy controls. Multivariate statistical analysis showed that the BD group was significantly distinguishable from the healthy control. Totally, 37 urinary metabolites responsible for discriminating BD subjects from healthy controls were identified. Interestingly, of 37 differential metabolites, 2,4-dihydroxypyrimidine was identified as an effective diagnostic biomarker for BD, yielding an area under the receiver operating characteristic curve (AUC) of 0.889 in the training samples (45 BD subjects and 61 healthy controls) and 0.805 in the test samples (26 BD subjects and 33 healthy controls). Our findings suggest that 2,4-dihydroxypyrimidine is a promising candidate urinary biomarker for BD, which may facilitate development of a urine-based diagnostic test for BD.


Assuntos
Transtorno Bipolar/diagnóstico , Transtorno Bipolar/urina , Pirimidinas/urina , Adulto , Biomarcadores/urina , Índice de Massa Corporal , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Masculino , Metaboloma , Metabolômica , Curva ROC
9.
Biomed Chromatogr ; 28(3): 446-52, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24254834

RESUMO

A sensitive and accurate HPLC-MS/MS method was developed for the simultaneous determination of dextromethorphan, dextrorphan and chlorphenamine in human plasma. Three analytes were extracted from plasma by liquid-liquid extraction using ethyl acetate and separated on a Kromasil 60-5CN column (3 µm, 2.1 × 150 mm) with mobile phase of acetonitrile-water (containing 0.1% formic acid; 50:50, v/v) at a flow rate of 0.2 mL/min. Quantification was performed on a triple quadrupole tandem mass spectrometer in multiple reaction monitoring mode using positive electrospray ionization. The calibration curve was linear over the range of 0.01-5 ng/mL for dextromethorphan, 0.02-5 ng/mL for dextrorphan and 0.025-20 ng/mL for chlorphenamine. The lower limits of quantification for dextromethorphan, dextrorphan and chlorphenamine were 0.01, 0.02 and 0.025 ng/mL, respectively. The intra- and inter-day precisions were within 11% and accuracies were in the range of 92.9-102.5%. All analytes were proved to be stable during sample storage, preparation and analytic procedures. This method was first applied to the pharmacokinetic study in healthy Chinese volunteers after a single oral dose of the formulation containing dextromethorphan hydrobromide (18 mg) and chlorpheniramine malaeate (8 mg).


Assuntos
Clorfeniramina/sangue , Cromatografia Líquida de Alta Pressão/métodos , Dextrometorfano/sangue , Dextrorfano/sangue , Espectrometria de Massas em Tandem/métodos , Adulto , Clorfeniramina/química , Clorfeniramina/farmacocinética , Dextrometorfano/química , Dextrometorfano/farmacocinética , Dextrorfano/química , Dextrorfano/farmacocinética , Estabilidade de Medicamentos , Feminino , Humanos , Análise dos Mínimos Quadrados , Masculino , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Espectrometria de Massas por Ionização por Electrospray/métodos , Adulto Jovem
10.
J Pharm Biomed Anal ; 88: 22-6, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24018419

RESUMO

A pressure-assisted capillary zone electrophoresis method was developed to determine the pKa values of a triptolide derivative (LLDT-246) and its impurities. The method was performed in an uncoated fused-silica capillary under the electric voltage of 18kV and 50mbar of external pressure applied simultaneously. A series of running electrolyte buffers were used with pH ranging between 2.2 and 10.0 with the constant ionic strength of 0.05M. The values of pKa of LLDT-246 and two impurities were calculated based on the pH dependence of effective mobilities (µeff). The pKa value of LLDT-246 was in good agreement with that of determined by potentiometric titration method.


Assuntos
Diterpenos/química , Eletroforese Capilar/métodos , Fenantrenos/química , Automação , Tampões (Química) , Química Farmacêutica , Diterpenos/análise , Desenho de Fármacos , Eletroquímica , Eletrólitos , Compostos de Epóxi/análise , Compostos de Epóxi/química , Concentração de Íons de Hidrogênio , Imunossupressores/análise , Íons , Modelos Químicos , Concentração Osmolar , Fenantrenos/análise , Extratos Vegetais/análise , Pressão , Tripterygium/metabolismo
11.
J Zhejiang Univ Sci B ; 14(11): 1004-12, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-24190446

RESUMO

A relationship between status epilepticus (SE) and oxidative stress has recently begun to be recognized. To explore whether the flavonoids extracted from licorice (LFs) have any protective effect on kainate (KA)-induced seizure in mice, we treated mice with LFs before and after KA injection. In KA-treated mice, we found that superoxide dismutase (SOD) activity decreased immediately after the onset of seizure at 1 h and then increased at 6 h. It returned to baseline 1 d after seizure and then increased again at 3, 7, and 28 d, while malondialdehyde (MDA) content remained at a high level at 1 h, 6 h, 3 d, 7 d, and 28 d, indicating a more oxidized status related to the presence of more reactive oxygen species (ROS). Treatment with LFs before KA injection reversed the seizure-induced change in SOD activity and MDA content at 1 h, 6 h, 3 d, 7 d, and 28 d. Treatment with LFs after seizure decreased KA-induced SOD activity and MDA content at 7 and 28 d. Also, LF pre- and post-KA treatments decreased seizure-induced neuronal cell death. Subsequently, Morris water maze tests revealed that the escape latency was significantly decreased and the number of target quadrant crossings was markedly increased in the LF-treated groups. Thus, our data indicate that LFs have protective effects on seizure-induced neuronal cell death and cognitive impairment through their anti-oxidative effects.


Assuntos
Antioxidantes/farmacologia , Flavonoides/farmacologia , Glycyrrhiza/química , Ácido Caínico/toxicidade , Fármacos Neuroprotetores/farmacologia , Estado Epiléptico/prevenção & controle , Animais , Transtornos Cognitivos/prevenção & controle , Masculino , Malondialdeído/análise , Camundongos , Camundongos Endogâmicos ICR , Estresse Oxidativo , Estado Epiléptico/induzido quimicamente , Superóxido Dismutase/metabolismo
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