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1.
Talanta ; 210: 120652, 2020 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-31987200

RESUMEN

Here, the novel petal-shaped ionic liquids modified covalent organic frameworks (PS-IL-COFs) particles have been synthesized by using ionic liquids as modifying agent, which could be beneficial to avoid the aggregation of COFs during the preparation and improve its dispersing performance. The novel PS-IL-COFs particles have been used and evaluated in the one step cleanup and extraction (OSCE) procedure for human plasma prior to the analysis of 3 general anesthetics by liquid chromatography-tandem quadrupole mass spectrometry (LC-MS/MS). In the OSCE procedure, human plasma samples are directly mixed with extraction solvent and PS-IL-COFs particles, and the extraction and cleanup procedure have been carried out simultaneously. Compared with the Oasis PRiME HLB cartridge method, the OSCE procedure using PS-IL-COFs particles as sorbents is much more effective for the minimization of ion suppression resulted from blood phospholipids. Under optimal conditions, the PS-IL-COFs particles show higher cleanup efficiency of 3 general anesthetics with recoveries in the range of 82.5%-115%. The limits of quantification (LOQs) for propofol, ketamine and etomidate are 0.18 µg/L, 0.15 µg/L and 0.016 µg/L, respectively. Validation results on linearity, specificity, precision and trueness, as well as on the application to analysis of general anesthetics in a case of a 54-year-old female suffered gallstone demonstrate the applicability to clinical studies.


Asunto(s)
Anestésicos/sangre , Etomidato/sangre , Líquidos Iónicos/química , Ketamina/sangre , Compuestos Orgánicos/química , Propofol/sangre , Cromatografía Líquida de Alta Presión , Femenino , Humanos , Persona de Mediana Edad , Tamaño de la Partícula , Propiedades de Superficie , Espectrometría de Masas en Tándem
2.
Braz J Med Biol Res ; 52(4): e7728, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-30994730

RESUMEN

Pituitary adenoma is one of the most common tumors in the neuroendocrine system. This study investigated the effects of long non-coding RNAs (lncRNAs) highly up-regulated in liver cancer (HULC) on rat secreting pituitary adenoma GH3 cell viability, migration, invasion, apoptosis, and hormone secretion, as well as the underlying potential mechanisms. Cell transfection and qRT-PCR were used to change and measure the expression levels of HULC, miR-130b, and FOXM1. Cell viability, migration, invasion, and apoptosis were assessed using trypan blue staining assay, MTT assay, two-chamber transwell assay, Guava Nexin assay, and western blotting. The concentrations of prolactin (PRL) and growth hormone (GH) in culture supernatant of GH3 cells were assessed using ELISA. The targeting relationship between miR-130b and FOXM1 was verified using dual luciferase activity. Finally, the expression levels of key factors involved in PI3K/AKT/mTOR and JAK1/STAT3 pathways were evaluated using western blotting. We found that HULC was highly expressed in GH3 cells. Overexpression of HULC promoted GH3 cell viability, migration, invasion, PRL and GH secretion, as well as activated PI3K/AKT/mTOR and JAK1/STAT3 pathways. Knockdown of HULC had opposite effects and induced cell apoptosis. HULC negatively regulated the expression of miR-130b, and miR-130b participated in the effects of HULC on GH3 cells. FOXM1 was a target gene of miR-130b, which was involved in the regulation of GH3 cell viability, migration, invasion, and apoptosis, as well as PI3K/AKT/mTOR and JAK1/STAT3 pathways. In conclusion, HULC tumor-promoting roles in secreting pituitary adenoma might be via down-regulating miR-130b, up-regulating FOXM1, and activating PI3K/AKT/mTOR and JAK1/STAT3 pathways.


Asunto(s)
Adenoma/patología , Neoplasias Hipofisarias/patología , ARN Largo no Codificante/fisiología , Adenoma/genética , Adenoma/metabolismo , Animales , Apoptosis/fisiología , Western Blotting , Línea Celular Tumoral , Ensayos de Migración Celular , Movimiento Celular/fisiología , Supervivencia Celular/fisiología , Ensayo de Inmunoadsorción Enzimática , Proteína Forkhead Box M1/análisis , Proteína Forkhead Box M1/metabolismo , Humanos , Janus Quinasa 1/análisis , Janus Quinasa 1/metabolismo , Luciferasas , MicroARNs/análisis , MicroARNs/metabolismo , Fosfatidilinositol 3-Quinasas/análisis , Fosfatidilinositol 3-Quinasas/metabolismo , Neoplasias Hipofisarias/genética , Neoplasias Hipofisarias/metabolismo , Proteínas Proto-Oncogénicas c-akt/análisis , Proteínas Proto-Oncogénicas c-akt/metabolismo , ARN Largo no Codificante/análisis , Ratas , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factor de Transcripción STAT3/análisis , Factor de Transcripción STAT3/metabolismo , Serina-Treonina Quinasas TOR/análisis , Serina-Treonina Quinasas TOR/metabolismo , Transfección
3.
Braz. j. med. biol. res ; 52(4): e7728, 2019. graf
Artículo en Inglés | LILACS | ID: biblio-1001506

RESUMEN

Pituitary adenoma is one of the most common tumors in the neuroendocrine system. This study investigated the effects of long non-coding RNAs (lncRNAs) highly up-regulated in liver cancer (HULC) on rat secreting pituitary adenoma GH3 cell viability, migration, invasion, apoptosis, and hormone secretion, as well as the underlying potential mechanisms. Cell transfection and qRT-PCR were used to change and measure the expression levels of HULC, miR-130b, and FOXM1. Cell viability, migration, invasion, and apoptosis were assessed using trypan blue staining assay, MTT assay, two-chamber transwell assay, Guava Nexin assay, and western blotting. The concentrations of prolactin (PRL) and growth hormone (GH) in culture supernatant of GH3 cells were assessed using ELISA. The targeting relationship between miR-130b and FOXM1 was verified using dual luciferase activity. Finally, the expression levels of key factors involved in PI3K/AKT/mTOR and JAK1/STAT3 pathways were evaluated using western blotting. We found that HULC was highly expressed in GH3 cells. Overexpression of HULC promoted GH3 cell viability, migration, invasion, PRL and GH secretion, as well as activated PI3K/AKT/mTOR and JAK1/STAT3 pathways. Knockdown of HULC had opposite effects and induced cell apoptosis. HULC negatively regulated the expression of miR-130b, and miR-130b participated in the effects of HULC on GH3 cells. FOXM1 was a target gene of miR-130b, which was involved in the regulation of GH3 cell viability, migration, invasion, and apoptosis, as well as PI3K/AKT/mTOR and JAK1/STAT3 pathways. In conclusion, HULC tumor-promoting roles in secreting pituitary adenoma might be via down-regulating miR-130b, up-regulating FOXM1, and activating PI3K/AKT/mTOR and JAK1/STAT3 pathways.


Asunto(s)
Humanos , Animales , Ratas , Neoplasias Hipofisarias/patología , Adenoma/patología , ARN Largo no Codificante/fisiología , Ensayo de Inmunoadsorción Enzimática , Transfección , Adenoma/genética , Adenoma/metabolismo , Movimiento Celular/fisiología , Supervivencia Celular/fisiología , Western Blotting , Apoptosis/fisiología , MicroARNs/análisis , Línea Celular Tumoral , Factor de Transcripción STAT3/análisis , Janus Quinasa 1/análisis , Janus Quinasa 1/metabolismo , Ensayos de Migración Celular , Proteína Forkhead Box M1/análisis , Proteína Forkhead Box M1/metabolismo , Luciferasas
4.
Anal Chim Acta ; 1020: 41-50, 2018 Aug 22.
Artículo en Inglés | MEDLINE | ID: mdl-29655427

RESUMEN

An enhanced cleanup efficiency hydroxy functionalized-magnetic graphene oxide (EH-Mag-GO) fully covered porous nano-titania as coating has been designed and synthesized. It has been evaluated in PRiME (process, robustness, improvements, matrix effects, ease of use) pass-through cleanup procedure for human plasma prior to analysis of strychnine and brucine by liquid chromatography-tandem quadrupole mass spectrometry (LC-MS/MS). Comparing with the magnetic carboxyl-graphene (Mag-CG), EH-Mag-GO is much more effective for the removal of matrix effect resulted from blood phospholipids. Under optimal conditions, the results show higher cleanup efficiency of EH-Mag-GO with recoveries in the range of 89.4%-118%. The limits of quantification (LOQs) for strychnine and brucine are 0.088 µg/L and 0.092 µg/L, respectively. Especially, the EH-Mag-GO is also evaluated for reuse (20 times) without much sacrifice of the cleanup efficiency. Validation results on linearity, specificity, accuracy and precision, as well as on the application to analysis of strychnine and brucine in six cases of suspected semen strychni poisoning demonstrate the applicability to clinical studies.


Asunto(s)
Grafito/química , Óxidos/química , Estricnina/análogos & derivados , Estricnina/sangre , Cromatografía Liquida , Humanos , Fenómenos Magnéticos , Conformación Molecular , Tamaño de la Partícula , Propiedades de Superficie , Espectrometría de Masas en Tándem
5.
Anal Chim Acta ; 960: 72-80, 2017 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-28193364

RESUMEN

A novel PRiME (process, robustness, improvements, matrix effects, ease of use) pass-through cleanup procedure has been developed to improve the existing commercially available designs. Carbon nanosorbents, i.e., magnetic modified carboxyl-graphene (Mag-CG) and magnetic modified carboxyl-carbon nanotubes (Mag-CCNTs), have been synthesised and evaluated in PRiME pass-through cleanup procedure for human plasma prior to analysis of 10 selected local anesthetic drugs by liquid chromatography-tandem quadrupole mass spectrometry (LC-MS/MS). The matrix effect, an interesting phenomenon of ion suppression for local anesthetic drugs containing ester group and ion enhancement for other drugs containing acylamino group, has been minimized using carbon nanosorbents PRiME pass-through cleanup procedure. Under the optimal conditions, the obtained results show higher cleanup efficiency of the carbon nanosorbents with recoveries between 70.2% and 126%. Furthermore, the carbon nanosorbents are also evaluated for reuse up to 80-100 times. The limits of quantification (LOQs) for local anesthetic drugs are in the range of 0.024-0.15 µg/L. Validation results on linearity, specificity, accuracy, and precision, as well as the application to the analysis of lidocaine in five patients recruited from the lung cancer demonstrate the applicability to clinical studies.


Asunto(s)
Anestésicos Locales/sangre , Anestésicos Locales/química , Análisis Químico de la Sangre/métodos , Nanotubos de Carbono/química , Grafito/química , Humanos , Imanes/química , Espectrometría de Masas en Tándem
6.
J Chromatogr A ; 1429: 86-96, 2016 Jan 15.
Artículo en Inglés | MEDLINE | ID: mdl-26718182

RESUMEN

A novel, simple and sensitive method based on the use of dispersive micro-solid-phase extraction (d-µ-SPE) procedure combined with ultra-fast liquid chromatography-tandem quadrupole mass spectrometry (UFLC-MS/MS) for the determination of catecholamines, i.e., dopamine (DA), norepinephrine(NE) and epinephrine (E), was developed and validated. The novel catecholamines molecularly imprinted polymer (MIP) on the surface of carboxyl-functionalized magnetic-carbon nanotube (CF@m-CNTs-MIP) was synthesized and characterized by vibrating sample magnetometer (VSM), scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS). The CF@m-CNTs-MIP was used as the d-µ-SPE sorbent to extract catecholamines from human plasma samples. The obtained results demonstrated the higher extraction capacity of CF@m-CNTs-MIP with recoveries between 87.5-110%. The limits of quantification (LOQs) for NE, E and DA were 76 ng/L, 18 ng/L and 10 ng/L, respectively. Validation results on linearity, specificity, accuracy, precision and stability, as well as on application to the analysis of catecholamines in 120 healthy volunteers demonstrated the applicability to clinical studies.


Asunto(s)
Análisis Químico de la Sangre/instrumentación , Análisis Químico de la Sangre/métodos , Catecolaminas/sangre , Cromatografía Liquida , Nanotubos de Carbono/química , Polímeros/química , Espectrometría de Masas en Tándem , Humanos , Magnetismo , Extracción en Fase Sólida
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