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1.
Mol Med ; 30(1): 77, 2024 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-38840035

RESUMO

BACKGROUND: Ischemic stroke presents a significant threat to human health due to its high disability rate and mortality. Currently, the clinical treatment drug, rt-PA, has a narrow therapeutic window and carries a high risk of bleeding. There is an urgent need to find new effective therapeutic drugs for ischemic stroke. Icariin (ICA), a key ingredient in the traditional Chinese medicine Epimedium, undergoes metabolism in vivo to produce Icaritin (ICT). While ICA has been reported to inhibit neuronal apoptosis after cerebral ischemia-reperfusion (I/R), yet its underlying mechanism remains unclear. METHODS: PC-12 cells were treated with 200 µM H2O2 for 8 h to establish a vitro model of oxidative damage. After administration of ICT, cell viability was detected by Thiazolyl blue tetrazolium Bromide (MTT) assay, reactive oxygen species (ROS) and apoptosis level, mPTP status and mitochondrial membrane potential (MMP) were detected by flow cytometry and immunofluorescence. Apoptosis and mitochondrial permeability transition pore (mPTP) related proteins were assessed by Western blotting. Middle cerebral artery occlusion (MCAO) model was used to establish I/R injury in vivo. After the treatment of ICA, the neurological function was scored by ZeaLonga socres; the infarct volume was observed by 2,3,5-Triphenyltetrazolium chloride (TTC) staining; HE and Nissl staining were used to detect the pathological state of the ischemic cortex; the expression changes of mPTP and apoptosis related proteins were detected by Western blotting. RESULTS: In vitro: ICT effectively improved H2O2-induced oxidative injury through decreasing the ROS level, inhibiting mPTP opening and apoptosis. In addition, the protective effects of ICT were not enhanced when it was co-treated with mPTP inhibitor Cyclosporin A (CsA), but reversed when combined with mPTP activator Lonidamine (LND). In vivo: Rats after MCAO shown cortical infarct volume of 32-40%, severe neurological impairment, while mPTP opening and apoptosis were obviously increased. Those damage caused was improved by the administration of ICA and CsA. CONCLUSIONS: ICA improves cerebral ischemia-reperfusion injury by inhibiting mPTP opening, making it a potential candidate drug for the treatment of ischemic stroke.


Assuntos
Apoptose , Flavonoides , AVC Isquêmico , Potencial da Membrana Mitocondrial , Poro de Transição de Permeabilidade Mitocondrial , Estresse Oxidativo , Espécies Reativas de Oxigênio , Animais , Estresse Oxidativo/efeitos dos fármacos , Ratos , Flavonoides/farmacologia , Flavonoides/uso terapêutico , Poro de Transição de Permeabilidade Mitocondrial/metabolismo , Apoptose/efeitos dos fármacos , AVC Isquêmico/tratamento farmacológico , AVC Isquêmico/metabolismo , AVC Isquêmico/etiologia , Células PC12 , Espécies Reativas de Oxigênio/metabolismo , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Masculino , Traumatismo por Reperfusão/metabolismo , Traumatismo por Reperfusão/tratamento farmacológico , Modelos Animais de Doenças , Peróxido de Hidrogênio/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Proteínas de Transporte da Membrana Mitocondrial/metabolismo , Fármacos Neuroprotetores/farmacologia , Fármacos Neuroprotetores/uso terapêutico , Ratos Sprague-Dawley
2.
Rev Med Virol ; 33(2): e2425, 2023 03.
Artigo em Inglês | MEDLINE | ID: mdl-36683235

RESUMO

Dengue illness can range from mild illness to life-threatening haemorrhage. It is an Aedes-borne infectious disease caused by the dengue virus, which has four serotypes. Each serotype acts as an independent infectious agent. The antibodies against one serotype confer homotypic immunity but temporary protection against heterotypic infection. Dengue has become a growing health concern for up to one third of the world's population. Currently, there is no potent anti-dengue medicine, and treatment for severe dengue relies on intravenous fluid management and pain medications. The burden of dengue dramatically increases despite advances in vector control measures. These factors underscore the need for a vaccine. Various dengue vaccine strategies have been demonstrated, that is, live attenuated vaccine, inactivated vaccine, DNA vaccine, subunit vaccine, and viral-vector vaccines, some of which are at the stage of clinical testing. Unfortunately, the forefront candidate vaccine is less than satisfactory, and its performance depends on serostatus and age factors. The lessons from clinical studies depicted ambiguity concerning the efficacy of dengue vaccine. Our study highlighted that viral structural heterogeneity, epitope accessibility, autoimmune complications, genetic variants, genetic diversities, antigen competition, virulence variation, host-pathogen specific interaction, antibody-dependent enhancement, cross-reactive immunity among Flaviviruses, and host-susceptibility determinants not only influence infection outcomes but also hampered successful vaccine development. This review integrates dengue determinants allocated necessities and challenges, which would provide insight for universal dengue vaccine development.


Assuntos
Vacinas contra Dengue , Vírus da Dengue , Vacinas Virais , Animais , Humanos , Anticorpos Antivirais , Mosquitos Vetores , Desenvolvimento de Vacinas
3.
Proc Natl Acad Sci U S A ; 118(8)2021 02 23.
Artigo em Inglês | MEDLINE | ID: mdl-33593916

RESUMO

The TGFß cytokine family member, GDF-15, reduces food intake and body weight and represents a potential treatment for obesity. Because the brainstem-restricted expression pattern of its receptor, GDNF Family Receptor α-like (GFRAL), presents an exciting opportunity to understand mechanisms of action for area postrema neurons in food intake; we generated GfralCre and conditional GfralCreERT mice to visualize and manipulate GFRAL neurons. We found infection or pathophysiologic states (rather than meal ingestion) stimulate GFRAL neurons. TRAP-Seq analysis of GFRAL neurons revealed their expression of a wide range of neurotransmitters and neuropeptides. Artificially activating GfralCre -expressing neurons inhibited feeding, decreased gastric emptying, and promoted a conditioned taste aversion (CTA). GFRAL neurons most strongly innervate the parabrachial nucleus (PBN), where they target CGRP-expressing (CGRPPBN) neurons. Silencing CGRPPBN neurons abrogated the aversive and anorexic effects of GDF-15. These findings suggest that GFRAL neurons link non-meal-associated pathophysiologic signals to suppress nutrient uptake and absorption.


Assuntos
Aprendizagem da Esquiva/efeitos dos fármacos , Ingestão de Alimentos/efeitos dos fármacos , Comportamento Alimentar/efeitos dos fármacos , Receptores de Fator Neurotrófico Derivado de Linhagem de Célula Glial/metabolismo , Fator 15 de Diferenciação de Crescimento/farmacologia , Neurônios/fisiologia , Núcleos Parabraquiais/fisiologia , Animais , Peso Corporal , Feminino , Receptores de Fator Neurotrófico Derivado de Linhagem de Célula Glial/genética , Masculino , Camundongos , Neurônios/efeitos dos fármacos , Núcleos Parabraquiais/efeitos dos fármacos , Ratos , Ratos Long-Evans
5.
Anim Biotechnol ; 34(8): 4041-4049, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37671949

RESUMO

Background: Yak is the main livestock species in the plateau area, and its reproductive performance is low, usually two years or three years. A very few of yaks recover within a certain period of time after delivery and smoothly enter the next estrous cycle, while most of them enter the postpartum anestrus and show no estrus performance. However, the key biological factors and influencing mechanisms that cause postpartum anestrus in yaks are not clear. Objective: To study the expression of differential transcripts in ovaries of yak during pregnancy and postpartum anestrus. Methods: Each three yaks in pregnancy and anestrus under natural grazing conditions in Haiyan County, Qinghai Province were selected and slaughtered, and their ovaries were collected and sent to Biomarker Technologies. Oxford Nanopore Technologies single-molecule real-time electrical signal sequencing technology was used to perform full-length transcriptome sequencing. Astalavista software was used to identify the types of alternative splicing events in yak estrus and pregnancy, and TAPIS pipeline was used to identify alternative polyadenylation. Results: The results showed that there were 1751 differentially expressed transcripts (DETs) between pregnancy and anestrus in yak, of which 808 were upregulated and 943 were downregulated. GO analysis showed that the biological processes of DETs were mainly reproductive, reproductive and rhythmic processes. KEGG analysis showed that the DET cell junction-related adhesion junction protein (ß-catenin) and amino terminal kinase (JNK) were involved in FAs (local adhesion). Phosphatidylinositol-3-kinase (PI3K) is involved in the PI3K/AKT/mTOR signaling pathway. Circadian rhythm output cycle failure (Clock) and brain and muscle tissue aromatic hydrocarbon receptor nuclear transporter-like protein 1 (Bmal1) are involved in circadian rhythm signaling pathway. Conclusion: This study found that ß-catenin, JNK, PI3K, Clock and Bmal1 were closely related to postpartum anestrus in yak.


Assuntos
Anestro , beta Catenina , Gravidez , Feminino , Bovinos/genética , Animais , Anestro/fisiologia , Fatores de Transcrição ARNTL , Fosfatidilinositol 3-Quinases , Período Pós-Parto/fisiologia
6.
Reprod Domest Anim ; 57(3): 292-303, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-34850471

RESUMO

Post-partum ovarian cycle arrest is the main factor affecting yak reproductive efficiency. There are few reports regarding the molecular regulatory mechanism of post-partum oestrus at transcriptome and proteome levels in yaks. Our previous studies focussed on the ovaries of yaks with post-partum ovarian cycle arrest and post-partum oestrus yaks. In this study, RNA sequencing transcriptomic study was combined with quantitative proteomic analyses to identify post-partum ovarian cycle-related genes and proteins. Consequently, 1,149 genes and 24 proteins were found to be up- or downregulated during post-partum oestrus. The analysis of differentially regulated genes identified three gene or protein pairs that were synchronously upregulated and no gene or protein pairs that were synchronously downregulated, suggesting that these upregulated genes may regulate the post-partum ovarian cycle. The functional classification of these differentially expressed genes and proteins indicated their connection with the oocyte meiosis, the oestrogen signalling pathway, the progesterone-mediated oocyte maturation and the gonadotrophin-releasing hormone (GnRH) signalling pathway. In this study, a total of six genes and two proteins involved in the oocyte meiosis, the oestrogen signalling pathway, the progesterone-mediated oocyte maturation and the GnRH signalling pathway were identified. The CSNK1A1, M91_09723, M91_11326, M91_21439, M91_19073, SHC2, Atf6b, M91_03062, HSPCA and calmodulin could regulate oestrus, respectively, in the post-partum so as to control the anoestrus status.


Assuntos
Proteômica , Transcriptoma , Animais , Bovinos , Feminino , Ciclo Menstrual , Ovário/metabolismo , Período Pós-Parto
7.
FASEB J ; 34(11): 14863-14877, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-32918517

RESUMO

Appropriate control of hepatic gluconeogenesis is essential for the organismal survival upon prolonged fasting and maintaining systemic homeostasis under metabolic stress. Here, we show protein arginine methyltransferase 1 (PRMT1), a key enzyme that catalyzes the protein arginine methylation process, particularly the isoform encoded by Prmt1 variant 2 (PRMT1V2), is critical in regulating gluconeogenesis in the liver. Liver-specific deletion of Prmt1 reduced gluconeogenic capacity in cultured hepatocytes and in the liver. Prmt1v2 was expressed at a higher level compared to Prmt1v1 in hepatic tissue and cells. Gain-of-function of PRMT1V2 clearly activated the gluconeogenic program in hepatocytes via interactions with PGC1α, a key transcriptional coactivator regulating gluconeogenesis, enhancing its activity via arginine methylation, while no effects of PRMT1V1 were observed. Similar stimulatory effects of PRMT1V2 in controlling gluconeogenesis were observed in human HepG2 cells. PRMT1, specifically PRMT1V2, was stabilized in fasted liver and hepatocytes treated with glucagon, in a PGC1α-dependent manner. PRMT1, particularly Prmt1v2, was significantly induced in the liver of streptozocin-induced type 1 diabetes and high fat diet-induced type 2 diabetes mouse models and liver-specific Prmt1 deficiency drastically ameliorated diabetic hyperglycemia. These findings reveal that PRMT1 modulates gluconeogenesis and mediates glucose homeostasis under physiological and pathological conditions, suggesting that deeper understanding how PRMT1 contributes to the coordinated efforts in glycemic control may ultimately present novel therapeutic strategies that counteracts hyperglycemia in disease settings.


Assuntos
Gluconeogênese , Hepatócitos/metabolismo , Hiperglicemia/genética , Proteína-Arginina N-Metiltransferases/metabolismo , Animais , Células Cultivadas , Mutação com Ganho de Função , Glucagon/metabolismo , Glucose/metabolismo , Células Hep G2 , Humanos , Hiperglicemia/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo/metabolismo , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Proteína-Arginina N-Metiltransferases/genética
8.
Analyst ; 146(6): 1986-1995, 2021 Mar 21.
Artigo em Inglês | MEDLINE | ID: mdl-33502393

RESUMO

Aptamers, which are called chemical antibodies for their high affinity and specificity to targets, have great potential as analytical tools to detect pesticides. In this work, a DNA aptamer for thiamethoxam was isolated by an improved SELEX (systematic evolution of ligands by exponential enrichment) strategy, in which the ssDNA library was fixed on streptavidin-agarose beads through a short biotin labeled complementary strand. After 13 rounds of selection, the random ssDNA pool was successfully enriched. Three sequences were chosen as aptamer candidates through sequencing and analysis and were transformed into fluorescent probes to evaluate their interactions with thiamethoxam. A fluorescent turn-on aptasensor for thiamethoxam based on the best aptamer (FAM-Thi13) and a short quenching strand were further designed and showed a quantitative linear range from 10 to 1000 nM with a detection limit of 1.23 nM for thiamethoxam. Molecular docking and molecular dynamics were used to investigate the binding site of the main probe of the aptasensor (FAM-Thi13) and thiamethoxam. Satisfactory results were also obtained in quantifying thiamethoxam in environmental water samples by the developed fluorescent aptasensor.


Assuntos
Aptâmeros de Nucleotídeos , Praguicidas , DNA de Cadeia Simples/genética , Simulação de Acoplamento Molecular , Técnica de Seleção de Aptâmeros , Tiametoxam
9.
J Am Chem Soc ; 142(23): 10331-10336, 2020 Jun 10.
Artigo em Inglês | MEDLINE | ID: mdl-32475113

RESUMO

The two-dimensional thin metal-organic frameworks (MOF) sheet has emerged as a promising hybrid material for applications in catalysis and optoelectronic devices. However, the small size and large thickness of an MOF sheet still pose barriers toward its potential applications. Herein, a micron-sized ultrathin MOF sheet is synthesized with the assistance of benzoic acid. Benzoic acid promoted the coordination of the porphyrin center with copper ions, reduced H-stacking and J-aggregation between the layers, and induced anisotropic growth of the MOF sheet. The results reveal the growth mechanism and provide a viable method for the synthesis of ultrathin MOF sheet. The as-prepared micron-sized ultrathin MOF sheet has good dispersion and high stability, which can ensure the long-term application properties of this material. The ultrathin thickness in combination with its micron size can make MOF as useful as graphene in practical applications. The synthesis of a micron-sized ultrathin MOF sheet similar to the thickness of graphene can pave the way for effective applications of two-dimensional MOF materials.

10.
J Clin Pharm Ther ; 45(6): 1434-1441, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32860258

RESUMO

WHAT IS KNOWN AND OBJECTIVE: Hypofibrinogenaemia is major treatment-related adverse event associated with tigecycline therapy, which in some cases can result in treatment termination. We aimed to identify the risk factors for tigecycline-induced hypofibrinogenaemia. METHODS: We retrospectively retrieved 426 Chinese patients who were undergoing tigecycline therapy ≥ 3 days. RESULTS AND DISCUSSION: There were 426 patients treated with tigecycline. The mean age was 60.31 ± 19.23 years, and 299 (70.19%) patients were male. Of the patients, 50.5% developed hypofibrinogenaemia and 10.1% of patients developed bleeding. Compared with before treatment, fibrinogen (FIB) significantly decreased after tigecycline was used while prothrombin time (PT), activated partial thromboplastin time (APTT) and thrombin time (TT) significantly increased (all P < .001). There was no statistically significant difference in platelet count, hepatic function, and renal function before and after tigecycline treatment (all P > .05). In analysing relevant risk factors, extension of the tigecycline treatment course was found to be the main risk factor for tigecycline-induced hypofibrinogenaemia. Regardless of whether patients received the standard dose or high dose of tigecycline, the long treatment course group (>14 days) had more patients with hypofibrinogenaemia than the routine treatment course group (52.21% vs 40.74%, 48.81% vs 19.44%, all P < .05). Renal failure (whether requiring or not requiring dialysis) is also a risk factor for tigecycline-induced hypofibrinogenaemia (OR [95% CI]: 2.450 [1.335-4.496]). WHAT IS NEW AND CONCLUSION: Tigecycline administration has been related to hypofibrinogenaemia, especially patients with renal failure and when long treatment course of tigecycline are used. We recommend that coagulation function be closely monitored in patients with the aforementioned risk factors for tigecycline-induced hypofibrinogenaemia to ensure patient safety.


Assuntos
Afibrinogenemia/induzido quimicamente , Antibacterianos/efeitos adversos , Tigeciclina/efeitos adversos , Adulto , Afibrinogenemia/epidemiologia , Idoso , Antibacterianos/administração & dosagem , Feminino , Hemorragia/induzido quimicamente , Hemorragia/epidemiologia , Humanos , Masculino , Pessoa de Meia-Idade , Tempo de Tromboplastina Parcial , Tempo de Protrombina , Insuficiência Renal/complicações , Estudos Retrospectivos , Fatores de Risco , Tempo de Trombina , Tigeciclina/administração & dosagem , Fatores de Tempo
11.
J Mol Evol ; 87(7-8): 231-239, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31432230

RESUMO

In this study, a new strategy for the selection of aptamers against small-molecule target was established using gold nanoparticles (AuNPs) as the separation matrix and Zinc(II)-Protoporphyrin IX (ZnPPIX) as the target molecule without the immobilization step due to the absorption of ssDNA on AuNPs. The progress of the selection process was monitored by the recovery rate and the fluorescence enhancement of N-methyl mesoporphyrin IX (NMM) after reacting with each selected pool. After 11 rounds of selection, a truncated aptamer ZnP1.2 with a low-micromolar dissociation constant was obtained, and it also showed good fluorescence enhancement for NMM and the enhanced peroxidase activity after binding with hemin, indicating this functional aptamer has potential to be a light-up fluorescent probe and a DNAzyme which could be used as an alternative to peroxidases for many colorimetric or chemiluminescent detections in biosensing events. The experimental results show that the simple and convenient AuNP-based SELEX is very conducive to the selection of aptamers for small-molecule targets.


Assuntos
Aptâmeros de Nucleotídeos/genética , Porfirinas/genética , Técnica de Seleção de Aptâmeros/métodos , DNA Catalítico/metabolismo , DNA de Cadeia Simples , Ouro/química , Nanopartículas Metálicas/química , Porfirinas/metabolismo , Protoporfirinas/química , Zinco/química
12.
J Mol Evol ; 87(2-3): 72-82, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30659315

RESUMO

Aptamers have become the most promising recognition reagents in terms of early diagnosis and effective treatment of cancers. In this study, using cervical cancer as a model, we have identified a DNA aptamer specifically binding to cervical cancer cells with high affinity using the cell-SELEX (systematic evolution of ligands by exponential enrichment) method, in which a negative selection was carried out using normal epithelial cells as control. The binding abilities of 6 selected truncated aptamers were determined by laser confocal fluorescence microscopy and flow cytometry, while most of them only recognize the target cells and do not bind the control cells, and the aptamer C-9S with 51-mer shows the best binding affinity to Ca Ski cells (target cells) with a dissociation constant value of 19.3 ± 2.9 nM. Moreover, at physiological temperature, C-9S remains its specific recognition capability to Ca Ski cells as well. Meanwhile, C-9S shows a similar binding ability to another cervical cancer cells (HeLa). Therefore, on the basis of its excellent targeting properties and inherent functional versatility of aptamer, C-9S holds great potential to be a molecular probe for early detection, in vivo imaging, and targeted delivery for further researches in cancer.


Assuntos
Técnica de Seleção de Aptâmeros/métodos , Neoplasias do Colo do Útero/diagnóstico por imagem , Neoplasias do Colo do Útero/genética , Aptâmeros de Nucleotídeos/metabolismo , Linhagem Celular Tumoral , DNA de Cadeia Simples/genética , Feminino , Citometria de Fluxo/métodos , Humanos , Ligantes , Microscopia Confocal/métodos , Sondas Moleculares/genética , Neoplasias do Colo do Útero/diagnóstico
13.
Bioorg Med Chem ; 27(20): 114969, 2019 10 15.
Artigo em Inglês | MEDLINE | ID: mdl-31262665

RESUMO

Computer-aided drug design has advanced by leaps and bounds, and has been widely used in various fields, and especially in the field of drug discovery. Although the crystal structure of the gibberellin (GA) receptor GID1A had been reported in previous studies, there is still a lack of designs of gibberellin functional analogue based GID1A. In the present study, a series of 30 thiourea derivatives were designed, synthesized and biologically assayed. The results suggested that the synthetic compounds had good GA-like activities. Furthermore, the structure-activity relationship of the synthetic compounds was discussed, and the dynamic simulation and docking study revealed the binding properties of the GID1A receptor and compounds Y1, Y11, and Y21.


Assuntos
Arabidopsis/efeitos dos fármacos , Desenho de Fármacos , Giberelinas/farmacologia , Simulação de Dinâmica Molecular , Tioureia/farmacologia , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Relação Dose-Resposta a Droga , Giberelinas/química , Simulação de Acoplamento Molecular , Estrutura Molecular , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/metabolismo , Relação Estrutura-Atividade , Tioureia/análogos & derivados , Tioureia/química
14.
Stroke ; 49(7): 1610-1617, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-29895539

RESUMO

BACKGROUND AND PURPOSE: Blood pressure (BP) control in the early phase of stroke is controversial to reduce the risk of poststroke cognitive impairment (PSCI). This study was to investigate the impact of BP levels in the early phase of ischemic stroke and stroke subtype on PSCI. METHODS: Seven hundred and ninety-six patients with acute ischemic stroke were included. Cognitive function was assessed after stroke onset using the Montreal Cognitive Assessment. Patients were divided into quintiles according to systolic BP and diastolic BP levels in the early phase. Subtype analyses were according to Trial of ORG 10172 in Acute Stroke Treatment classification (infarct cause) and Oxfordshire Community Stroke Project classification (infarct location). RESULTS: After adjusting for multiple variables, the quintiles with the lowest systolic BP (Q1, 102-127 mm Hg) and with the highest systolic BP (Q5, 171-215 mm Hg) were associated with increased PSCI risk (odds ratio, 1.83; 95% confidence interval, 1.64-2.28; P=0.007 in Q1; odds ratio, 2.32; 95% confidence interval, 1.74-2.90; P<0.001 in Q5) at 3 months as compared with the middle quintile (Q3, 143-158 mm Hg). Similar association was found in diastolic BP quintiles. The analysis of cerebral infarction subtype demonstrated that both large artery atherosclerosis and total anterior circulation infarct were associated with increased risk of PSCI at 3 months after adjusting for multiple variables (large artery atherosclerosis: odds ratio, 1.42; 95% confidence interval, 1.06-1.90; P=0.031; total anterior circulation infarct: odds ratio, 1.68; 95% confidence interval, 1.32-2.15; P=0.001). CONCLUSIONS: Lower or higher BP in the early phase of ischemic stroke was correlated with increased PSCI risk at 3 months. Maintaining systolic/diastolic BP in the levels of 143 to 158/93 to 102 mm Hg might be beneficial to reduce the occurrence of PSCI. Moreover, large artery atherosclerosis subtype and total anterior circulation infarct subtype were correlated with increased PSCI risk at 3 months. CLINICAL TRIAL REGISTRATION: URL: https://www.chictr.org. Unique identifier: ChiCTR-TRC-14004804.


Assuntos
Pressão Sanguínea/fisiologia , Isquemia Encefálica/complicações , Disfunção Cognitiva/etiologia , Acidente Vascular Cerebral/complicações , Idoso , Determinação da Pressão Arterial , Isquemia Encefálica/fisiopatologia , Disfunção Cognitiva/fisiopatologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Testes Neuropsicológicos , Fatores de Risco , Acidente Vascular Cerebral/fisiopatologia
15.
Photochem Photobiol Sci ; 17(6): 800-806, 2018 Jun 13.
Artigo em Inglês | MEDLINE | ID: mdl-29770378

RESUMO

Aptamers are short single-stranded DNA or RNA, which can be selected in vitro by systematic evolution of ligands by exponential enrichment (SELEX). In order to develop novel light-up probes to substitute G-quadruplex (G4), we selected a DNA aptamer for crystal violet (CV), a triphenylmethane light-up dye, by a modified affinity chromatography-based SELEX. The ssDNA pool was first coupled on streptavidin-coated agarose beads through a biotin labeled complementary oligonucleotide, and then the aptamer sequences would be released from agarose beads by CV affinity. This method is simple, straightforward and effective. The aptamer sequence with a low micromolar dissociation constant (Kd) and good specificity was achieved after 11 rounds of selection. The light-up properties of the CV-aptamer were also investigated, and the CV showed dramatic fluorescence enhancement. The CV-aptamer pair could be further used as a novel light-up fluorescent probe to design biosensors.

16.
Nanomedicine ; 14(7): 2387-2395, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-28552643

RESUMO

The low efficacy and high toxicity of chemotherapy have been driving increasing attention on development of combined anticancer therapy technique. In the current work, graphene oxide (GO)-hybridized nanogels (AGD) were developed for delivery of an anticancer drug (doxorubicin (DOX)), which simultaneously presented photothermal therapeutic effects against cancer cells. AGD nanogels were fabricated by in situ incorporating GO nanoplatelets into a biodegradable polymer (alginate) via a double emulsion approach using a disulfide molecule as crosslinker, followed by DOX encapsulation via electrostatic interactions. The nanogels released DOX drug in an accelerated way under both acidic and reducible conditions mimicking extracellular tumor microenvironments and intracellular compartments. The stimulative release controllability of the nanogels improved the DOX internalization and long-term drug accumulation inside A549 cells (an adenocarcinoma human alveolar basal epithelial cell line), which, together with their photothermal effect, resulted in a good anticancer cytotoxicity, indicating their promising potential for combinative anticancer therapy.


Assuntos
Antibióticos Antineoplásicos/administração & dosagem , Doxorrubicina/administração & dosagem , Sistemas de Liberação de Medicamentos , Grafite/química , Neoplasias Pulmonares/tratamento farmacológico , Nanopartículas/administração & dosagem , Antibióticos Antineoplásicos/química , Sobrevivência Celular , Doxorrubicina/química , Humanos , Neoplasias Pulmonares/patologia , Nanopartículas/química , Células Tumorais Cultivadas
17.
Small ; 12(36): 5090-5097, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-27445096

RESUMO

Over the last decade, significant progress has been made to develop sensitive devices for the capture of circulating tumor cells (CTCs) from blood of cancer patients. However, simple capture and counting of CTCs cannot provide effective information for understanding the biology of them. In this work, a functional biointerface is fabricated for specific capture and nondestructive release of CTCs from blood samples. A nanostrucure of porous network based on chitosan nanofibers is fabricated by electrospinning, to mimic the function of extracellular matrices, and then the poly(carboxybetaine methacrylate) (pCBMA) brushes integrating onto nanofiber interface provide the effect of interfacial properties to control nonspecific cell adhesion and the multivalent immobilization of aptamers to induce high efficient and specific CTC capture. Furthermore, a complementary sequence is used to efficiently hybridize with the aptamer to achieve nondestructive release of the captured target cells, assisted by the flexible space provided by pCBMA brushes. This work also shows how nanostructure and the interface molecules regulate the morphology of the captured CTCs, and reveals the importance of the controllable cell morphology on biointerface for an effective nondestructive release of the captured CTCs.


Assuntos
Betaína/química , Quitosana/química , Nanofibras/química , Células Neoplásicas Circulantes/patologia , Ácidos Polimetacrílicos/química , Linhagem Celular , Forma Celular , Fluorescência , Humanos , Nanofibras/ultraestrutura
18.
Anal Bioanal Chem ; 408(3): 683-93, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26577084

RESUMO

Doxorubicin (Dox) is a DNA-targeting anthracycline antibiotic active against a wide spectrum of cancers. The interaction between Dox and double-stranded DNA (dsDNA) was used to load Dox using DNA duplexes as carriers. More importantly, the interesting DNA sequence-dependent fluorescence response of Dox could be exploited in the design of efficient Dox release systems and efficient fluorescence sensors. In this work, we demonstrated that separate introduction of G and C bases into T-rich single-stranded DNA (ssDNA) sequences afforded the best discrimination of Dox binding between dsDNA and ssDNA. For the first time, we successfully utilized this interesting DNA sequence-dependent fluorescence response of Dox as a signal transduction mechanism for the sensitive detection of biothiols in human serum. Cysteine, homocysteine, and glutathione were detected at as low as 26 nM, 37 nM, and 29 nM, respectively. The biosensors exhibited not only good selectivity, stability, and sensitivity in aqueous solutions but also a sensitive response in human serum, demonstrating their potential for diagnosis.


Assuntos
Antibióticos Antineoplásicos/química , Técnicas Biossensoriais/métodos , DNA de Cadeia Simples/química , DNA/química , Doxorrubicina/química , Compostos de Sulfidrila/sangue , Antraciclinas/química , Antibióticos Antineoplásicos/administração & dosagem , Antibióticos Antineoplásicos/sangue , Doxorrubicina/administração & dosagem , Doxorrubicina/sangue , Sistemas de Liberação de Medicamentos , Fluorescência , Humanos
19.
Mol Biol Rep ; 43(11): 1227-1232, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27558095

RESUMO

The fibrillin-1 (FBN1) gene mutations result in Marfan syndrome (MFS) and have a variety of phenotypic variations. This disease is involved in the skeletal, ocular and cardiovascular system. Here we analyzed genotype-phenotype correlation in two Chinese families with MFS. Two patients with thoracic aortic aneurysms and dissections were diagnosed as MFS according to the revised Ghent criteria. Peripheral blood samples were collected and genomic DNAs were isolated from available cases, namely, patient-1 and his daughter and son, and patient-2 and his parents. According to the next-generation sequencing results, the mutations in FBN1 were confirmed by direct sequencing. A heterozygous frameshift mutation in exon 12 of FBN1 was found in the proband-1 and his daughter. They showed cardiovascular phenotype thoracic aortic aneurysms and dissections, a life-threatening vascular disease, and atrial septal defect respectively. One de novo missense mutation in exon 50 of FBN1 was identified only in the patient-2, showing aortic root aneurysm and aortic root dilatation. Intriguingly, two novel mutations mainly caused the cardiovascular complications in affected family members. No meaningful mutations were found in these two patients by screening all exons of 428 genes related with cardiovascular disease. The high incidence of cardiovascular manifestations might be associated with the two novel mutations in exon 12 and 50 of FBN1.


Assuntos
Aneurisma da Aorta Torácica/genética , Dissecção Aórtica/genética , Povo Asiático/genética , Fibrilina-1/genética , Síndrome de Marfan/genética , Adolescente , Adulto , China , Éxons , Feminino , Estudos de Associação Genética , Predisposição Genética para Doença , Humanos , Masculino , Mutação , Linhagem , Análise de Sequência de DNA , Adulto Jovem
20.
Small ; 11(40): 5444-51, 2015 Oct 28.
Artigo em Inglês | MEDLINE | ID: mdl-26313660

RESUMO

Circulating tumor cell (CTC) isolation has attracted a great deal of research interest in recent years. However, there are still some challenges, including purity as well as viability of the captured CTCs, resulting from nanoscale structures and inorganic nanomaterials. Here, a chitosan nanoparticle surface is first fabricated by electrospray to provide a cellular compatible interface. The "soft" substrate, further modified by polyethylene glycol (PEG) as an antifouling molecule and DNA aptamer as a specific capture molecule, has a hydrophilic nature and is capable of specific capture of viable rare CTCs from artificial white blood cell (WBC) samples. Furthermore, a subsequent in situ culture strategy based on the developed cellular compatible soft interface is introduced for further purification and proliferation of the captured rare number target cells. The WBCs are weeded out after 2 d, and after a 7 d proliferation nearly 200 MCF-7 cells are obtained from 7 target cells with more than 90% purity. This work provides a promising strategy for viable isolation and purification of rare CTCs and it has great potential for achieving clinical validity.


Assuntos
Técnicas de Cultura de Células/métodos , Separação Celular/métodos , Quitosana/química , Nanoestruturas/química , Células Neoplásicas Circulantes , Humanos , Células MCF-7 , Nanopartículas/química
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