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BACKGROUND: Influenza A viruses (IAV) are extremely common respiratory viruses for the acute exacerbation of chronic obstructive pulmonary disease (AECOPD), in which IAV infection may further evoke abnormal macrophage polarization, amplify cytokine storms. Melatonin exerts potential effects of anti-inflammation and anti-IAV infection, while its effects on IAV infection-induced AECOPD are poorly understood. METHODS: COPD mice models were established through cigarette smoke exposure for consecutive 24 weeks, evaluated by the detection of lung function. AECOPD mice models were established through the intratracheal atomization of influenza A/H3N2 stocks in COPD mice, and were injected intraperitoneally with melatonin (Mel). Then, The polarization of alveolar macrophages (AMs) was assayed by flow cytometry of bronchoalveolar lavage (BAL) cells. In vitro, the effects of melatonin on macrophage polarization were analyzed in IAV-infected Cigarette smoking extract (CSE)-stimulated Raw264.7 macrophages. Moreover, the roles of the melatonin receptors (MTs) in regulating macrophage polarization and apoptosis were determined using MTs antagonist luzindole. RESULTS: The present results demonstrated that IAV/H3N2 infection deteriorated lung function (reduced FEV20,50/FVC), exacerbated lung damages in COPD mice with higher dual polarization of AMs. Melatonin therapy improved airflow limitation and lung damages of AECOPD mice by decreasing IAV nucleoprotein (IAV-NP) protein levels and the M1 polarization of pulmonary macrophages. Furthermore, in CSE-stimulated Raw264.7 cells, IAV infection further promoted the dual polarization of macrophages accompanied with decreased MT1 expression. Melatonin decreased STAT1 phosphorylation, the levels of M1 markers and IAV-NP via MTs reflected by the addition of luzindole. Recombinant IL-1ß attenuated the inhibitory effects of melatonin on IAV infection and STAT1-driven M1 polarization, while its converting enzyme inhibitor VX765 potentiated the inhibitory effects of melatonin on them. Moreover, melatonin inhibited IAV infection-induced apoptosis by suppressing IL-1ß/STAT1 signaling via MTs. CONCLUSIONS: These findings suggested that melatonin inhibited IAV infection, improved lung function and lung damages of AECOPD via suppressing IL-1ß/STAT1-driven macrophage M1 polarization and apoptosis in a MTs-dependent manner. Melatonin may be considered as a potential therapeutic agent for influenza virus infection-induced AECOPD.
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Apoptosis , Subtipo H3N2 del Virus de la Influenza A , Melatonina , Enfermedad Pulmonar Obstructiva Crónica , Animales , Melatonina/farmacología , Enfermedad Pulmonar Obstructiva Crónica/tratamiento farmacológico , Enfermedad Pulmonar Obstructiva Crónica/metabolismo , Enfermedad Pulmonar Obstructiva Crónica/virología , Enfermedad Pulmonar Obstructiva Crónica/fisiopatología , Ratones , Apoptosis/efectos de los fármacos , Células RAW 264.7 , Subtipo H3N2 del Virus de la Influenza A/efectos de los fármacos , Infecciones por Orthomyxoviridae/tratamiento farmacológico , Infecciones por Orthomyxoviridae/metabolismo , Infecciones por Orthomyxoviridae/inmunología , Ratones Endogámicos C57BL , Masculino , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Progresión de la Enfermedad , Polaridad Celular/efectos de los fármacos , Modelos Animales de Enfermedad , Macrófagos Alveolares/efectos de los fármacos , Macrófagos Alveolares/metabolismo , Macrófagos Alveolares/virologíaRESUMEN
PURPOSE: Cytokeratin 19-positive cancer stem cells (CK19 + CSCs) and their tumor-associated macrophages (TAMs) have not been fully explored yet in the hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC). EXPERIMENTAL DESIGN: Single-cell RNA sequencing was performed on the viable cells obtained from 11 treatment-naïve HBV-associated HCC patients, including 8 CK19 + patients, to elucidate their transcriptomic landscape, CK19 + CSC heterogeneity, and immune microenvironment. Two in-house primary HCC cohorts (96 cases-related HBV and 89 cases with recurrence), TCGA external cohort, and in vitro and in vivo experiments were used to validate the results. RESULTS: A total of 64,581 single cells derived from the human HCC and adjacent normal tissues were sequenced, and 11 cell types were identified. The result showed that CK19 + CSCs were phenotypically and transcriptionally heterogeneous, co-expressed multiple hepatics CSC markers, and were positively correlated with worse prognosis. Moreover, the SPP1 + TAMs (TAM_SPP1) with strong M2-like features and worse prognosis were specifically enriched in the CK19 + HCC and promoted tumor invasion and metastasis by activating angiogenesis. Importantly, matrix metalloproteinase 9 (MMP9) derived from TAM_SPP1, as the hub gene of CK19 + HCC, was activated by the VEGFA signal. CONCLUSIONS: This study revealed the heterogeneity and stemness characteristics of CK19 + CSCs and specific immunosuppressive TAM_SPP1 in CK19 + HCC. The VEGFA signal can activate TAM_SPP1-derived MMP9 to promote the invasion and metastasis of CK19 + HCC tumors. This might provide novel insights into the clinical treatment of HCC patients.
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Carcinoma Hepatocelular , Neoplasias Hepáticas , Humanos , Carcinoma Hepatocelular/patología , Neoplasias Hepáticas/patología , Virus de la Hepatitis B/genética , Metaloproteinasa 9 de la Matriz/genética , Queratina-19/genética , Queratina-19/metabolismo , Macrófagos Asociados a Tumores/metabolismo , Macrófagos Asociados a Tumores/patología , Células Madre Neoplásicas , Análisis de Secuencia de ARN , Microambiente Tumoral , Osteopontina/genética , Osteopontina/metabolismoRESUMEN
BACKGROUND: The application of vonoprazan significantly improved the eradication rate of Helicobacter pylori (H. pylori). This study aimed to compare efficacy and safety of the 10-day vonoprazan-amoxicillin (VA) and 14-day rabeprazole-amoxicillin (RA) dual therapy, and to provide a more efficient, safer, and convenient dual regimen for H. pylori infection. METHODS: This was a prospective, open-label, multi-center, randomized controlled study of treatment-naive patients with H. pylori infection. The participants were randomly assigned to the 10-day VA group with vonoprazan 20 mg Bid plus amoxicillin 1 g Tid or the 14-day RA group with rabeprazole 10 mg Tid plus amoxicillin 1 g Tid. The effectiveness, the adverse events, and the patient compliance of the two groups were compared. RESULTS: A total of 690 patients were enrolled. The eradication rates of 10-day VA and 14-day RA dual therapy were 89.3% and 84.9% in intention-to-treat (ITT) analysis (P = 0.088); 90.6% and 85.9% by modified intention-to-treat (mITT) analysis (P = 0.059); 91.4% and 86.6% by per-protocol (PP) analysis (P = 0.047). Non-inferiority was confirmed between the two groups (all P < 0.001). No discernible differences were observed in adverse effects and compliance between groups. Poor compliance reduced the eradication efficacy (P < 0.05). CONCLUSIONS: The 10-day VA dual therapy was non-inferior to the 14-day RA dual therapy for H. pylori treatment-naive patients, which should be given priority in the first-line treatment. The application of vonoprazan reduced treatment course and antibiotic use. Patients' adherence was crucial for the success of eradication.
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Infecciones por Helicobacter , Helicobacter pylori , Humanos , Infecciones por Helicobacter/tratamiento farmacológico , Rabeprazol/efectos adversos , Estudios Prospectivos , Inhibidores de la Bomba de Protones/efectos adversos , Claritromicina/uso terapéutico , Quimioterapia Combinada , Antibacterianos/efectos adversos , Amoxicilina/efectos adversos , Resultado del TratamientoRESUMEN
BACKGROUND: Traditional bismuth-containing quadruple therapy, as a first-line eradication treatment for Helicobacter pylori (H. pylori), has several disadvantages, including drug side effects, low medication adherence, and high costs. Trials of high-dose dual treatment have demonstrated its advantages, which include good safety and adherence profiles. In this study, we investigated the efficacy, safety, and compliance of a high-dose dual therapy when compared with bismuth-based quadruple treatment for the initial eradication of H. pylori infection on Hainan Island, China. METHODS: We randomized 846 H. pylori-infected patients into two groups. A bismuth-containing quadruple therapy group was administered the following: esomeprazole 20 mg, amoxicillin 1000 mg, and clarithromycin 500 mg twice daily, and colloidal bismuth pectin in suspension 150 mg three times/day for 2 weeks. A high-dose dual therapy group was administered the following: esomeprazole 20 mg four times/day and amoxicillin 1000 mg three times/day for 2 weeks. Patients were given a 13C urea breath test at 4 weeks at treatment end. Adverse effects and compliance were evaluated at follow-up visits. RESULTS: Eradication rates in the high-dose dual therapy group were: 90.3% (381/422, 95% confidence interval [CI]: 87.1%-92.9%) in intention-to-treat (ITT) and 93.6% (381/407, 95% CI: 90.8%-95.8%) in per-protocol (PP) analyses. Eradication rates were 87.3% in ITT (370/424, 95% CI: 83.7%-90.3%) and 91.8% in PP analyses (370/403, 95% CI: 88.7%-94.3%) for quadruple therapy, with no statistical differences (P = 0.164 in ITT and P = 0.324 in PP analyses). Adverse effects were 13.5% (55/407) in the dual group and 17.4% (70/403) in the quadruple group (P = 0.129). Compliance was 92.4% (376/407) in the dual group and 86.6% (349/403) in the quadruple group (P = 0.007). CONCLUSIONS: High-dose dual therapy had high eradication rates comparable with bismuth-based quadruple treatment, with no differences in adverse effects, however higher adherence rates were recorded.
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Infecciones por Helicobacter , Helicobacter pylori , Humanos , Infecciones por Helicobacter/tratamiento farmacológico , Infecciones por Helicobacter/etiología , Bismuto/uso terapéutico , Bismuto/efectos adversos , Antibacterianos , Esomeprazol , Quimioterapia Combinada , Amoxicilina/efectos adversos , Resultado del Tratamiento , Inhibidores de la Bomba de Protones/efectos adversosRESUMEN
OBJECTIVE: To evaluate whether electroacupuncture (EA) would improve gastrointestinal function and clinical prognosis in patients with severe traumatic brain injury (TBI) complicocted by acute gastrointestinal injury (AGI). METHODS: This multicenter, single-blind trial included patients with TBI and AGI admitted to 5 Chinese hospitals from September 2018 to December 2019. A total of 500 patients were randomized to the control or acupuncture groups using a random number table, 250 cases in each group. Patients in the control group received conventional treatment, including mannitol, nutritional support, epilepsy and infection prevention, and maintenance of water, electrolytes, and acid-base balance. While patients in the acupuncture group received EA intervention at bilateral Zusanli (ST 36), Shangjuxu (ST 37), Xiajuxu (ST 39), Tianshu (ST 25), and Zhongwan (RN 12) acupoints in addition to the conventional treatment, 30 min per time, twice daily, for 7 d. The primary endpoint was 28-d mortality. The secondary endpoints were serum levels of D-lactic acid (D-lac), diamine oxidase (DAO), lipopolysaccharide (LPS), motilin (MTL) and gastrin (GAS), intra-abdominal pressure (IAP), bowel sounds, abdominal circumference, AGI grade, scores of gastrointestinal failure (GIF), Glasgow Coma Scale (GCS), Acute Physiology and Chronic Health Evaluation (APACHE II), Sequential Organ Failure Assessment (SOFA), and Multiple Organ Dysfunction Syndrome (MODS), mechanical ventilation time, intense care unit (ICU) stay, and the incidence of hospital-acquired pneumonia. RESULTS: The 28-d mortality in the acupuncture group was lower than that in the control group (22.80% vs. 33.20%, P<0.05). Compared with the control group, the acupuncture group at 7 d showed lower GIF, APACHE II, SOFA, MODS scores, D-lac, DAO, LPS, IAP, and abdominal circumference and higher GCS score, MTL, GAS, and bowel sound frequency (all P<0.05). In addition, the above indices showed simillar changes at 7 d compared with days 1 and 3 (all P<0.05) in the EA group. CONCLUSION: Early EA can improve gastrointestinal function and clinical prognosis in patients with severe TBI complicated by AGI. (Registration No. ChiCTR2000032276).
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Terapia por Acupuntura , Lesiones Traumáticas del Encéfalo , Electroacupuntura , Humanos , Lipopolisacáridos , Método Simple Ciego , Lesiones Traumáticas del Encéfalo/terapiaRESUMEN
BACKGROUND: MicroRNA-21 (miR-21) is related to hypertension and cardiac remodelling. Left atrium (LA) dilation is highly sensitive to small haemodynamic changes in the left ventricle (LV) that are induced by hypertension. This study aimed to elucidate the relationship between miR-21 expression and LA dilation in elderly patients with essential hypertension (EH). METHODS: In this cross-sectional study, one hundred elderly patients with EH were recruited for the study. According to their left atrium diameters (LADs), the patients were divided into the LA dilation group [42 patients (42%)] and the no-LA dilation group [58 patients (58%)]. The serum levels of miR-21 and chemical biomarkers used in the clinic, such as creatinine, blood urea nitrogen, uric acid, fasting blood glucose, total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), very-low-density lipoprotein cholesterol, Lp(a), apolipoprotein A1 (apoA1), and apolipoprotein B, were measured. All the patients underwent echocardiographic examination, and the LAD, interventricular septum (IVS), right atrium diameter (RAD), right ventricle diameter (RVD), left ventricular end-systolic diameter (LVESD), left ventricular end-systolic diameter (LVEDD) and left ventricular ejection fraction (LVEF) were measured. RESULTS: The levels of miR-21 [8.02 (5.21, 14.39) vs. 6.05 (3.81, 8.95), P = 0.011] and LVEF (67.02 ± 3.82 vs. 64.14 ± 4.43, P = 0.001) were higher in the LA dilation group. The levels of creatinine [70.40 (64.45, 80.15) vs. 63.9(60.1, 73.43)], P = 0.020] were higher in the no-LA dilation group. The levels of HDLC (r = - 0.209, P = 0.037), apoA1 (r = -0.269, P = 0.007) and RAD (r = 0.203, P = 0.043) were significantly correlated with miR-21 expression. The LAD was significantly correlated with the RAD (r = 0.287, P = 0.004), RVD (r = 0.450, P < 0.001), LVEDD (r = 0.248, P = 0.013) and LVEF (r = 0.232, P = 0.020). Multivariate logistic regression revealed that miR-21 significantly influenced LA dilation in elderly patients with EH (P < 0.05). CONCLUSIONS: Circulating serum levels of miR-21 are increased in elderly patients with EH with LA dilation. miR-21 levels are significantly correlated with LA dilation in elderly patients with EH, and miR-21 may be a factor related to the clinical pathophysiological occurrence of and treatment for the progression of hypertension-related early heart damage in EH patients.
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BACKGROUND & AIMS: The multiplicity of hepatocellular carcinoma (HCC) recurrence patterns is the most important determinant of patients' postsurgical survival. A systematic HCC recurrence classification is needed to help prevent and treat postoperative HCC recurrence in the era of precision medicine. METHODS: A total of 1319 patients with recurrent HCC from four hospitals were enrolled and divided into a development cohort (n = 916), internal validation cohort (n = 225) and external validation cohort (n = 178). A comprehensive study of patients' clinicopathological factors and biological features was conducted. RESULTS: Four subtypes of recurrence were identified, which integrated recurrence features, survival, effects on systemic and liver function and potential therapeutics after recurrence: type I (solitary-intrahepatic oligorecurrence); type II (multi-intrahepatic oligorecurrence); type III (progression recurrence) and type IV (hyper-progression recurrence). Type III~IV recurrence indicated exceptionally poor prognosis. Subsequently, two nomogram models were established for type III~IV recurrence prediction, and both demonstrated excellent predictive performance and applicability of pre and postoperative strategy formulation. Multiple biological analyses revealed that HCC cases with type III~IV recurrence were characterized by enrichment in p53 mutations, CCND1 amplification, high proliferation/metastasis potential, inactive metabolism and immune exhaustion features. Over-expression of high mobility group protein 2 (HMGA2) enhanced the highly malignant behaviour of HCC through multiple molecular pathways, making it a potential prognostic predictor and therapeutic target. CONCLUSIONS: This 'recurrent HCC classification' has important potential value in identifying patients with surgical benefit, predicting postsurgical survival and guiding treatment strategies. Multidimensional biological insights also increased knowledge of factors associated with HCC recurrence.
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Carcinoma Hepatocelular , Neoplasias Hepáticas , Carcinoma Hepatocelular/patología , Hepatectomía/métodos , Humanos , Neoplasias Hepáticas/patología , Recurrencia Local de Neoplasia/patología , Nomogramas , PronósticoRESUMEN
A simple and sensitive in situ antioxidation process assisted with a matrix solid-phase dispersion method for extracting chiral flavonoids in citrus fruit was established, and samples were further analyzed using ion mobility quadrupole time-of-flight high-resolution mass spectrometry. The collision cross-sections of the target compounds were studied using single-field and stepped-field methods. The optimal conditions were obtained using 30 mg of C18 as a dispersant, methanol as an elution solvent and 0.6 mM 1,1-diphenyl-2-picrylhydrazyl (DPPH) as a radical solution. Additionally, the method showed satisfactory limits of detection (3.70-6.52 ng/mL) and good recoveries (96.78-104.67%) for four flavonoids in citrus fruit. The IC50 values of DPPH radical-scavenging activities ranged from 817.8 to 981.55 µg/mL for tested samples. The method was a good alternative for the microextraction and determination of antioxidant capacity and chiral differentiation of narirutin, naringin, hesperidin and neohesperidin in citrus fruit.
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Citrus/química , Flavonoides/análisis , Análisis de los Alimentos/métodos , Espectrometría de Masas/métodos , Microextracción en Fase Sólida/métodos , Antioxidantes/química , Cromatografía Líquida de Alta Presión/métodos , Disacáridos/análisis , Flavanonas/análisis , Frutas/química , Hesperidina/análogos & derivados , Hesperidina/análisis , Espectrometría de Movilidad Iónica , Metanol/química , Solventes/químicaRESUMEN
The zirconium metal-organic framework (Zr-MOF) was used as a novel and effective adsorbent material for the enrichment of five phenylurea herbicides (fenuron, monuron, diuron, linuron and pencycuron) in natural products. The target analytes were determined by ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry. Some crucial experimental parameters, such as type of adsorbents, amount of adsorbent, type of eluent solvents and adsorption capacity were investigated and optimized. Under the optimum extraction conditions, the enrichment factors of fenuron, monuron, diuron, linuron and pencycuron were 90, 128, 148, 204 and 295 times, respectively. A good linearity was obtained in different concentration levels of target analytes with the determination coefï¬cients (r2) larger than 0.993. In addition, the limits of detection varied from 0.05 to 0.36â¯ng/mL and the recoveries of the analytes were in the range of 85.19-99.13 %. The results demonstrated that the proposed miniaturized solid-phase extraction procedure coupled with Zr-MOF could become an effective tool to analyze phenylurea herbicides and would have the vast application prospect for the extraction of pesticide residue and more organic pollutants from Hawthorn, Dendrobii Officinalis Caulis and Salviae Miltiorrhizae Radix et Rhizoma samples.
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Contaminantes Ambientales/análisis , Herbicidas/análisis , Estructuras Metalorgánicas/química , Residuos de Plaguicidas/análisis , Compuestos de Fenilurea/análisis , Circonio/química , Cromatografía Líquida de Alta Presión/métodos , Crataegus/química , Dendrobium/química , Espectrometría de Masas/métodos , Salvia miltiorrhiza/química , Extracción en Fase Sólida/métodosRESUMEN
A simple and effective vesicle based ultrasonic-assisted extraction (UAE) method was developed for extraction of active compounds in functional food. The target analytes were determined by ultra-high performance liquid chromatography with ultraviolet detector. Surfactant vesicle was adopted as extraction solvent. Different operating conditions including the type and concentration of vesicle, extraction time and solid to liquid ratio were investigated by single-factor experiments and response surface methodology. Optimized experimental conditions were 1% (w/v) of DTAB/SDS vesicle, 20â¯min of extraction time and 160â¯mg/mL of solid to liquid ratio. The proposed method provided good linearity in the linear range of 10-1000⯵g/mL with regression coefficients larger than 0.999, low limits of detection of 27.64-55.67â¯ng/mL, good precision with relative standard deviations below 0.35%, and satisfactory recoveries of 83.84-90.92% for tested saponins. Consequently, the proposed vesicle based UAE method was well suited for the extraction of saponins in Panax notoginseng.
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Fraccionamiento Químico/métodos , Panax notoginseng/química , Saponinas/aislamiento & purificación , Ultrasonido , Cromatografía Líquida de Alta Presión , Saponinas/análisisRESUMEN
A new bile acid tauro-16ß-hydroxy-12α-sulfate-5ß-cholenoic acid (1), along with six known ones (2-7), was isolated from the snake bile. Its planar structure and relative configuration were elucidated based on extensive spectroscopic analyses. Moreover, compound 2 showed inhibitory effect on NO production in RAW 264.7 macrophages at non-cytotoxic concentration (20 µM) with inhibitory rate of 69.7%. [Formula: see text].
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Bilis , Medicina Tradicional China , Ácidos y Sales Biliares , Estructura MolecularRESUMEN
A novel micro matrix solid phase dispersion method was successfully used for the extraction of quaternary alkaloids in Phellodendri chinensis cortex. The elution of target compounds was accomplished with sodium hexanesulfonate as the eluent solvent. A neutral ion pair was formed between ion-pairing reagent and positively charged alkaloids in this process, which was beneficial for selectively extraction of polar alkaloids. Several parameters were optimized and the optimal conditions were listed as follows: silica gel as the sorbent, silica to sample mass ratio of 1:1, the grinding time of 1 min. The exhaustive elution of targets was achieved by 200 µL methanol/water (9:1) containing 150 mM sodium hexane sulfonate at pH 4.5. The method validation covered linearity, recovery, precision of intraday and interday, limits of detection, limits of quantitation, and repeatability. This established method was rapid, simple, environmentally friendly, and highly sensitive.
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Berberina , Medicamentos Herbarios Chinos/química , Quinolizinas , Extracción en Fase Sólida/métodos , Berberina/análisis , Berberina/aislamiento & purificación , Cromatografía Líquida de Alta Presión , Límite de Detección , Modelos Lineales , Phellodendron/química , Plantas Medicinales/química , Quinolizinas/análisis , Quinolizinas/aislamiento & purificación , Reproducibilidad de los ResultadosRESUMEN
The quantification of three alkaloids is important because quantitative study is a means of assessing the reliability of the experimental method, and three alkaloids of peimine, peiminine, and peimisine are main active ingredients in Chinese Pharmacopoeia 2015. An effective method based on the matrix solid-phase dispersion microextraction was developed for the extraction of alkaloid compounds in Fritillariae Thunbergii Bulbus. Target analytes were analyzed by capillary electrophoresis coupled with quadrupole time-of-flight mass spectrometry. The optimized experimental condition was that 50 mg Fritillariae Thunbergii Bulbus was blended homogeneously with 10 mg citric acid for 5 min. Two hundred microliters of water acidized by 1 mol/L hydrochloric acid (pH = 4.5) was selected to elute tested alkaloids. The results demonstrated that the investigated method had low limits of detection (1.32-1.59 ng/mL), good recoveries (86.63-98.12%), and reproducibility (relative standard deviations of peak areas < 0.87%). The proposed matrix solid-phase dispersion microextraction coupled with capillary electrophoresis combined with quadrupole time-of-flight mass spectrometry was successfully applied for the extraction of alkaloids in plants.
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Alcaloides/análisis , Alcaloides/aislamiento & purificación , Medicamentos Herbarios Chinos/análisis , Medicamentos Herbarios Chinos/aislamiento & purificación , Electroforesis Capilar/métodos , Microextracción en Fase Sólida/métodos , Espectrometría de Masas en Tándem/métodos , Cevanas/análisis , Cevanas/aislamiento & purificación , Límite de DetecciónRESUMEN
INTRODUCTION: The main active components in hawthorn leaves possess various biological activities such as anti-inflammatory, antioxidant, and hypolipidemic effects. Therefore, it is necessary to develop an effective and reliable extraction method to extract these active compounds from hawthorn leaves. OBJECTIVE: To establish a simple, rapid, and sensitive method for extraction and determination of polyphenolic compounds from hawthorn leaves. METHODS: In this study, a microwave-assisted reaction and extraction (MARE) combined with ultra-high-performance liquid chromatography with ultraviolet detector method was established to extract and determine the polyphenolic compounds in hawthorn leaves. The solid reagent aqueous solutions were applied as extraction solvents, preventing the use of organic solvents. The target analytes were identified by quadrupole time-of-flight tandem mass spectrometry. Several experimental parameters that can significantly affect the extraction efficiency were evaluated and optimised. RESULTS: The optimal conditions were as follows: 0.1 g of sodium carbonate was used as solid reagent, the amount of sodium borate was set at 0.01 g, extraction time was 10 min, extraction temperature was set at 50°C, pH value was adjusted to 7. The validation experiments demonstrated that the method had high sensitivity with the limits of detection in the range 26.5-37.7 ng/mL. The average recoveries ranged from 80.22% to 93.27%. CONCLUSION: In this work, the proposed MARE method was successfully applied to extract and determine polyphenolic compounds in hawthorn leaf samples. Compared with other reported methods, the present method was faster, greener, and more sensitive.
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Crataegus/química , Microondas , Hojas de la Planta/química , Concentración de Iones de Hidrógeno , Límite de Detección , Polifenoles/análisis , Reproducibilidad de los ResultadosRESUMEN
A rapid and effective method was successfully established for the extraction and determination of chlorogenic acid, protocatechuic acid, malic acid, caffeic acid and gallic acid in fruit (chaenomeles speciosa) via matrix solid-phase dispersion (MSPD) microextraction combined with ultra-high performance liquid chromatography with quadrupole time-of-flight tandem mass spectrometry (UHPLC-Q-TOF/MS). Several major extraction parameters were investigated and optimized, such as the type of sorbent, the amount of sorbent, the grinding time, the type and concentration of the eluting solvent. The optimal extraction conditions were obtained by using 20 mg of calix[8]arene as dispersing adsorbent, selecting 60 s as the appropriate grinding time and applying 250 mM of 1-dodecyl-3-methylimidazolium bromide as eluent solvent. Moreover, the calibration curves of the analytes were in the range of 0.01-500 µg/mL with the determination coefficients (r2) higher than 0.9995. The limits of detection and limits of quantification were in the range of 0.202-1.056 ng/mL and 0.674-3.521 ng/mL, respectively. The recoveries of the target compounds at two spiked levels were between 82.19 and 113.36%. Furthermore, this method had acceptable reproducibility (RSD ≤ 3.84%). The proposed approach combined the advantages of MSPD microextraction with UHPLC-Q-TOF/MS, and could be applicable for the analysis organic acids in fruit.
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Calixarenos/química , Ácidos Carboxílicos/aislamiento & purificación , Frutas/química , Líquidos Iónicos/química , Microextracción en Fase Sólida/métodos , Adsorción , Iones/análisis , Límite de Detección , Modelos Lineales , Estándares de Referencia , Reproducibilidad de los Resultados , Solventes/química , Espectrometría de Masas en Tándem , Factores de TiempoRESUMEN
BACKGROUND/AIMS: Current practical advances in high-throughput data technologies including RNA-sequencing have led to the identification of long non-coding RNAs (lncRNAs) for potential clinical application against bladder urothelial cancer (BLCA). However, most previous studies focused on the clinical value of individual lncRNAs, which has limited the potential for future clinical application. METHODS: In this study, RNA-sequencing data of lncRNAs was downloaded from The Cancer Genome Atlas database. Risk score was constructed based on survival-associated lncRNAs identified using differential expression analysis as well as univariate and multivariate Cox proportional hazards regression analysis. Receiver operating characteristic and Kaplan-Meier curve analyses were employed to evaluate the clinical and prognostic value of risk scores. Bioinformatics analyses were used to investigate the potential mechanisms of newly identified lncRNAs. RESULTS: Among 2,127 differentially expressed lncRNAs (DELs), four new lncRNAs (AC145124.1, AC010168.2, MIR200CHG, and AC098613.1) showed valuable prognostic effects in BLCA patients. More importantly, the four-DEL-based risk score had the potential to become an independent marker for the survival status prediction of BLCA patients. Distinct co-expressed genes and signaling pathways were identified when BLCA was categorized into low- and high-risk groups. Furthermore, a protein-coding gene, HIST4H4 was found only 68 bp from the AC010168.2 DEL. HIST4H4 expression level was evidently up-regulated and positively correlated with AC010168.2 in BLCA patients. CONCLUSION: This in silico investigation pioneers the future investigation of the utility of prognostic lncRNAs for BLCA.
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ARN Largo no Codificante/metabolismo , Neoplasias de la Vejiga Urinaria/patología , Área Bajo la Curva , Biomarcadores de Tumor/genética , Biomarcadores de Tumor/metabolismo , Femenino , Regulación Neoplásica de la Expresión Génica , Redes Reguladoras de Genes , Humanos , Estimación de Kaplan-Meier , Pronóstico , Modelos de Riesgos Proporcionales , Curva ROC , Factores de Riesgo , Análisis de Secuencia de ARN , Neoplasias de la Vejiga Urinaria/genética , Neoplasias de la Vejiga Urinaria/mortalidadRESUMEN
BACKGROUND/AIMS: Increasing evidences indicated the important roles of alternative splicing in the progression and prognosis of bladder urothelial carcinoma (BLCA). However, most previous research has focused on one or several alternative splicing events, without a comprehensive evaluation of the prognostic value of splicing events in BLCA. In this study, we aimed to determine risk scores for predicting prognosis of BLCA patients based on splicing events. METHODS: RNA-sequencing data and clinical information of BLCA patients were downloaded from The Cancer Genome Atlas, and data of splicing events were obtained from the SpliceSeq database. Univariate and multivariate Cox regression analyses were employed to identify survival-associated alternative spicing events (SASEs) and to calculate risk scores. Protein-protein interaction analysis of genes of the SASEs was performed using STRING, a database of known and predicted protein-protein interactions, and pathway enrichment analysis of the genes was implemented using the Database for Annotation, Visualization and Integrated Discovery (version 6.8). Receiver operating characteristic (ROC) curves and Kaplan-Meier analysis were used to evaluate the clinical significance of genes from the SASEs for building a risk score in BLCA. Correlation between splicing events of splicing factors and non-splicing factors were analyzed with Pearson correlation coefficient. A potential regulatory network was then built using Cytoscape 3.5. RESULTS: In total, 39,508 alternative splicing events in 317 patients with BLCA were analyzed, including 4,632 SASEs. The area under the curve of the ROC of risk score (all) was 0.748 for predicting survival status of BLCA patients. Low- and high-risk score groups classified using the median "risk score (all)" value displayed remarkably different survival time (Low vs. High = 3304.841±239.758 vs 1198.614±152.460 days). The potential regulatory network with SASEs of splicing factors and other genes was constructed, which might be part of the biological mechanisms associated with prognosis of BLCA patients. CONCLUSIONS: In this study, prognostic signatures constructed using splicing events could be used for predicting the prognosis of BLCA patients.
Asunto(s)
Empalme Alternativo , Redes Reguladoras de Genes , ARN Mensajero/genética , Neoplasias de la Vejiga Urinaria/diagnóstico , Neoplasias de la Vejiga Urinaria/genética , Estudios de Cohortes , Femenino , Perfilación de la Expresión Génica , Regulación Neoplásica de la Expresión Génica , Humanos , Masculino , Pronóstico , Modelos de Riesgos ProporcionalesRESUMEN
A rapid, simple, and efficient sample extraction method based on micro-matrix-solid-phase dispersion (micro-MSPD) was applied to the extraction of polyphenols from pomegranate peel. Five target analytes were determined by ultra-HPLC coupled with Q-TOF/MS. Carbon molecular sieve (CMS) was firstly used as dispersant to improve extraction efficiency in micro-MSPD. The major micro-MSPD parameters, such as type of dispersant, amount of dispersant, grinding time, and the type and the volume of elution solvents, were studied and optimized. Under optimized conditions, 26 mg of pomegranate peel was dispersed with 32.5 mg of CMS, the grinding time was selected as 90 s, the dispersed sample was eluted with 100 µL of methanol. Results showed that the proposed method was of good linearity for concentrations of analytes against their peak areas (coefficient of determination r2 > 0.990), the LOD was as low as 3.2 ng/mL, and the spiking recoveries were between 88.1 and 106%. Satisfactory results were obtained for the extraction of gallic acid, punicalagin A, punicalagin B, catechin, and ellagic acid from pomegranate peel sample, which demonstrated nice reliability and high sensitivity of this approach.
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Cromatografía Líquida de Alta Presión/métodos , Lythraceae/química , Espectrometría de Masas/métodos , Polifenoles/aislamiento & purificación , Extracción en Fase Sólida/métodos , Carbono/química , Límite de Detección , Modelos Lineales , Extractos Vegetales/química , Polifenoles/análisis , Polifenoles/química , Reproducibilidad de los ResultadosRESUMEN
In this study, dispersive micro solid phase extraction (DMSPE) combined with ultrahigh-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry method was established to extract and determine sudan dyes and their metabolites in real samples. The crown ether microfunctionalized multi-walled carbon nanotubes (MWCNTs) was applied as the sorbent in DMSPE procedure. Several experimental parameters that can effect the extraction performance of the DMSPE method were investigated separately by a univariate method. The validation data showed that the limits of detection were in the range of 0.084-13.13⯵g/kg, the mean recoveries were ranged from 80.15 to 103.58% for six samples. Compared with other published methods, the proposed method was more effective, more time-saving and more eco-friendly. Finally, the developed method was successfully applied to enrich and detect sudan dyes and their metabolites in real samples.
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Compuestos Azo/aislamiento & purificación , Cromatografía Líquida de Alta Presión/métodos , Colorantes/aislamiento & purificación , Análisis de los Alimentos/métodos , Microextracción en Fase Sólida/métodos , Espectrometría de Masas en Tándem/métodos , Compuestos Azo/análisis , Colorantes/análisis , Éteres Corona/química , Límite de Detección , Nanotubos de Carbono/químicaRESUMEN
Mycotoxins are a group of secondary fungi metabolites present in foods that cause adverse effects in humans and animals. The objective of this study was to develop and validate a reliable and sensitive method to determine the presence of fumonisin B1, aflatoxin B1, ochratoxin B, T-2 toxin, ochratoxin A and zearalenone. A rapid, effective process, which involves microwave-assisted dispersive micro-solid phase extraction (MA-d-µ-SPE), has been proposed for the extraction and detection of 6 mycotoxins in peach seed, milk powder, corn flour and beer sample matrixes, for subsequent analysis by ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UHPLC-Q-TOF/MS). Several experimental parameters (type of dispersant, concentration of dispersant, vortex time, type of desorption solvent and pH) affecting the extraction efficiency were systematically studied and optimized. The optimum extraction conditions involved immersing 2.5⯵g/mL of nano zirconia (as dispersant) in a 5â¯mL sample solution. After 2â¯min of extraction by vigorous shaking, the target analytes were desorbed by 100⯵L of chloroform at pH 4.5. The results indicated good linearity in the range of 0.0074-3.6⯵g/mL (râ¯≥â¯0.9982), low limits of detection (0.0036-0.033⯵g/kg for solid samples and 0.0022-0.017â¯ng/mL for beer), acceptable reproducibility (relative standard deviation (RSD%) 2.08-2.76% for retention time and 3.51-4.59% for peak area, nâ¯=â¯3), and satisfactory spiked recoveries (84.27-104.96%) for studied mycotoxins in sample matrixes, which demonstrated that MA-d-µ-SPE coupled with UHPLC-Q-TOF/MS is a useful tool for analysis of multi-mycotoxin.