Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 102
Filtrar
Mais filtros










Base de dados
Intervalo de ano de publicação
1.
J Colloid Interface Sci ; 565: 483-493, 2020 Jan 09.
Artigo em Inglês | MEDLINE | ID: mdl-31982715

RESUMO

The complex biology of glioma compromises therapeutic efficacy and results in poor prognosis. Photodynamic therapy (PDT) has emerged as a promising modality for localized tumor ablation with limited damage to healthy brain tissues. However, low photosensitizer concentration and hypoxic microenvironment in glioma tissue hamper the practical applications of PDT. To address the challenges, biocompatible periodic mesoporous organosilica coated Prussian blue nanoparticles (PB@PMOs) are constructed to load a biosafe prodrug 5-aminolevulinic acid (5-ALA), which is pronouncedly converted to protoporphyrin IX (PpIX) in malignant cells. PB@PMO-5-ALA induces a higher accumulation of PpIX in glioma cells compared to free 5-ALA. Meanwhile, the PB@PMOs, with a mean edge length of 81 nm and good biocompatibility, effectively decompose hydrogen peroxide to oxygen in a temperature-responsive manner. Oxygen supply further contributes to the promotion of 5-ALA-PDT. Thus, the photodynamic effect of PB@PMO-5-ALA is significantly improved, imposing augmented cytotoxicity to glioma U87MG cells. Furthermore, ex vivo fluorescence imaging elucidates the tumor PpIX increases by 75% in PB@PMO-5-ALA treated mice than that in 5-ALA treated ones post 12 h injection. Magnetic resonance imaging (MRI) and iron staining strongly demonstrate the accumulation of PB@PMO-5-ALA in glioma tissues with negative contrast enhancement and blue staining deposits, respectively. The nanoparticle accumulation and high PpIX level collaboratively enhance PDT efficacy through PB@PMO-5-ALA, which efficiently suppresses tumor growth, providing a promising option with safety for local glioma ablation.

2.
Chemistry ; 2020 Jan 18.
Artigo em Inglês | MEDLINE | ID: mdl-31953964

RESUMO

"Click" chemistry is focused on the development of highly selective reactions utilizing simple precursors for the exquisite synthesis of molecules. Undisputedly, the Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) is one of the most valuable examples of click chemistry, but there are still some limitations as needing additional reducing agents and ligands as well as the toxicity of copper in cell or body. Here, we demonstrate a novel strategy of photoredox electron-transfer radical cation pattern for AAC reaction instead of traditional metal-catalyzed coordination process. This newly-developed photocatalyzed azide-alkyne cycloaddition (PcAAC) could be performed in mild conditions of room temperature, air and visible light with good functional group tolerance, excellent atom economy, high yield up to 99%, and absolute regioselectivity, affording variety of 1,4-triazole derivatives including bioactive molecules and pharmaceuticals. The use of a recyclable photocatalyst, solar energy and water makes this photocatalytic system sustainable and environmentally friendly. Moreover, the metal-free photocatalyzed AAC reaction with excellent regioselectivity was also implemented for the first time, representing an important development for click chemistry, which should find versatile applications in organic synthesis, chemical biology and materials science.

3.
Artif Cells Nanomed Biotechnol ; 48(1): 8-14, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31852248

RESUMO

Osteoarthritis is a common type of degenerative joint disease. Inflammation-related chondrocyte senescence plays a major role in the pathogenesis of osteoarthritis. Omentin-1 is a newly identified anti-inflammatory adipokine involved in lipid metabolism. In this study, we examined the biological function of omentin-1 in cultured chondrocytes. The presence of omentin-1 potently suppresses IL-1ß-induced cellular senescence as revealed by staining with senescence-associated beta-galactosidase (SA-ß-Gal). At the cellular level, omentin-1 attenuates IL-1ß-induced G1 phase cell-cycle arrest. Mechanistically, we demonstrate that omentin-1 reduced IL-1ß-induced expression of senescent factors including caveolin-1, p21, and PAI-1 as well as p53 acetylation through ameliorating SIRT1 reduction. Notably, silencing of SIRT1 abolishes IL-1ß-induced senescence along with the induction of p21 and PAI-1, suggesting that the action of omentin-1 in chondrocytes is dependent on SIRT1. Collectively, our results revealed the molecular mechanism through which the adipokine omentin-1 exerts a beneficial effect, thereby protecting chondrocytes from senescence. Thus, omentin-1 could have clinical implication in the treatment of osteoarthritis.

4.
Light Sci Appl ; 8: 109, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31798848

RESUMO

Two-photon laser scanning microscopy has been extensively applied to study in vivo neuronal activity at cellular and subcellular resolutions in mammalian brains. However, the extent of such studies is typically confined to a single functional region of the brain. Here, we demonstrate a novel technique, termed the multiarea two-photon real-time in vivo explorer (MATRIEX), that allows the user to target multiple functional brain regions distributed within a zone of up to 12 mm in diameter, each with a field of view (FOV) of ~200 µm in diameter, thus performing two-photon Ca2+ imaging with single-cell resolution in all of the regions simultaneously. For example, we demonstrate real-time functional imaging of single-neuron activities in the primary visual cortex, primary motor cortex and hippocampal CA1 region of mice in both anesthetized and awake states. A unique advantage of the MATRIEX technique is the configuration of multiple microscopic FOVs that are distributed in three-dimensional space over macroscopic distances (>1 mm) both laterally and axially but that are imaged by a single conventional laser scanning device. In particular, the MATRIEX technique can be effectively implemented as an add-on optical module for an existing conventional single-beam-scanning two-photon microscope without requiring any modification to the microscope itself. Thus, the MATRIEX technique can be readily applied to substantially facilitate the exploration of multiarea neuronal activity in vivo for studies of brain-wide neural circuit function with single-cell resolution.

5.
ACS Appl Mater Interfaces ; 11(50): 46626-46636, 2019 Dec 18.
Artigo em Inglês | MEDLINE | ID: mdl-31751121

RESUMO

Programmed cell death ligand 1 (PD-L1) blockade has achieved great success in cancer immunotherapy; however, the response of triple-negative breast cancer (TNBC) to PD-L1 antibodies is limited. To address this challenge, we use the bromodomain and extra-terminal inhibitor JQ1 to down-regulate the expression of PD-L1 and thus elicit the immune response to TNBC instead of using antibodies to block PD-L1. JQ1 also inhibits the growth of TNBC as a targeted therapeutic agent by inhibiting the BRD4-c-MYC axis. The polydopamine nanoparticles (PDMNs) are introduced as a biodegradable and adaptable platform to load JQ1 and induce photothermal therapy (PTT) as another synergistic therapeutic modality. Because the JQ1-loaded PDMNs (PDMN-JQ1) are self-degradable and release JQ1 continuously, this synergistic treatment can lead to remarkable activation of cytotoxic T lymphocytes and induce a strong immune-memory effect to protect mice from tumor re-challenge. Taken together, our study demonstrates a compact and simple nanoplatform for triple therapy, including targeted therapy, PTT, and immunotherapy, for TNBC treatment.

6.
J Biosci Bioeng ; 2019 Oct 28.
Artigo em Inglês | MEDLINE | ID: mdl-31672431

RESUMO

CotA-laccases are potential enzymes that are widely used in decolorization of dyes and degradation of toxic substances. In this study, a novel CotA-laccase gene from Bacillus pumilus W3 was applied for rational design. After a series of site-directed genetic mutations, the mutant S208G/F227A showed a 5.1-fold higher catalytic efficiency (kcat/Km) than the wild-type CotA-laccase did. The optimal pH of S208G/F227A was 3.5 with ABTS as substrate. The residual activity of mutant S208G/F227A was more than 80% after incubated for 10 h at pH 7-11. Mutant S208G/F227A showed optimal temperature at 80°C with ABTS as substrate. The thermal stability of mutant laccase S208G/F227A was lower than that of wild-type CotA-laccase. This study showed that Gly208 and Ala227 play key roles in catalytic efficiency and it is possible to improve catalytic efficiency of CotA-laccase through site-directed mutagenesis.

7.
PLoS One ; 14(8): e0219630, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31408462

RESUMO

Previous surveys of neonatal medicine in China have not collected comprehensive information on antibiotic use in newborns. The goal of the present study was to assess the trends in antibiotic use in inpatient newborns from advanced hospitals in mainland China and to evaluate the contributing factors. We extracted retrospective data on newborn clinical units from a database containing key clinical subspecialty area indicators from provincial or ministerial (Class A level III) hospitals over three consecutive years (2008-2010) and in 25 of 31 provincial districts of mainland China. Fifty-five newborn units were included in the study. The results showed that two thirds (65.7% ± 23.1%) of inpatient newborns were prescribed antibiotic products. Antibiotic use rates were significantly different by newborn ward bed capacity (p = 0.023; 60.6% for d capacity (ficant65.7% ± 23-100 beds group, and 77.1% for (ficant65.7% ± 23.1%) of inpatient newb significantly different by type of hospital, geographic area, admission to physician or nurse ratio, or physician or nurse academic degree. Factors contributing significantly to antibiotic use included ward bed capacity, physician to nurse ratio, average hospital stay, and pneumonia to preterm infant ratio. Our data suggested that the use of antibiotics among inpatient newborns in advanced hospitals in mainland China was prevalent and should be subject to rigorous monitoring, and highlighted the need to explore how newborn ward bed capacity potentially impacts antibiotic use.

8.
Front Neurosci ; 13: 663, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31293380

RESUMO

Understanding the complex neural mechanisms controlling urinary bladder activity is an extremely important topic in both neuroscience and urology. Simultaneously recording of the bladder activity and neural activity in related brain regions will largely advance this field. However, such recording approach has long been restricted to anesthetized animals, whose bladder function and urodynamic properties are largely affected by anesthetics. In our recent report, we found that it is feasible to record bladder pressure (cystometry) and the related cortical neuron activity simultaneously in freely moving mice. Here, we aimed to demonstrate the use of this combined method in freely moving mice for recording the activity of the pontine micturition center (PMC), a more difficultly approachable small region deeply located in the brainstem and a more popularly studied hub for controlling bladder function. Interestingly, we found that the duration of urination events linearly correlated to the time course of neuronal activity in the PMC. We observed that the activities of PMC neurons highly correlated with spike-like increases in bladder pressure, reflecting bladder contractions. We also found that anesthesia evoked prominent changes in the dynamics of the Ca2+ signals in the PMC during the bladder contraction and even induced the dripping overflow incontinence due to suppression of the neural activity in the PMC. In addition, we described in details both the system for cystometry in freely moving mice and the protocols for how to perform this combined method. Therefore, this work provides a powerful approach that enables the simultaneous measurement of neuronal activity of the PMC or any other brain sites and bladder function in freely behaving mice. This approach offers a promising possibility to examine the neural mechanisms underlying neurogenic bladder dysfunction.

9.
Cancer Lett ; 454: 146-157, 2019 Jul 10.
Artigo em Inglês | MEDLINE | ID: mdl-30978442

RESUMO

Metabolic abnormality is one of the hallmarks of cancer cells, and limiting material supply is a potential breakthrough approach for cancer treatment. Increasing researchers have been involved in the study of glioma cell metabolism reprogramming since the significance of IDH1 was confirmed in glioma. However, the molecular mechanisms underlying metabolic reprogramming induced by methionine deprivation regulates glioma cell proliferation remain unclear. Here we demonstrated that methionine deprivation inhibited glioma cell proliferation via downregulating interleukin 1 receptor antagonist (IL1RN) both in vitro and in vivo, methionine deprivation or knocking down IL1RN induced glioma cell cycle arrest. Moreover, we confirmed that IL1RN is a tumor associated gene and its expression is negatively correlated with the survival time of glioma patients. Altogether these results demonstrate a strong rationale insight that targeting amino acid metabolism such as methionine deprivation/IL1RN related gene therapy may offer novel direction for glioma treatment.

10.
Can J Physiol Pharmacol ; 97(3): 147-154, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30661367

RESUMO

Childhood obesity is a predictor of adult obesity and has its roots in the pre-pregnancy or pregnancy period. This review presents an overview of the prenatal risk factors for childhood obesity, which were categorized into 2 groups: biological risk factors (maternal pre-pregnancy body mass index, gestational weight gain, diabetes in pregnancy, and caesarean section), and environmental and behavioural risk factors (maternal smoking and exposure to obesogens, maternal dietary patterns, maternal intestinal microbiome and antibiotics exposure, and maternal psychosocial stress). Identifying modifiable predisposing prenatal factors for obesity will inform further development of inventions to prevent obesity over the life course, and future directions for research and intervention are discussed.


Assuntos
Obesidade Pediátrica/etiologia , Animais , Feminino , Humanos , Gravidez , Complicações na Gravidez/etiologia , Fatores de Risco
11.
Science ; 362(6413): 429-434, 2018 10 26.
Artigo em Inglês | MEDLINE | ID: mdl-30361367

RESUMO

Clinical observations indicate that the paramedian region of the thalamus is a critical node for controlling wakefulness. However, the specific nucleus and neural circuitry for this function remain unknown. Using in vivo fiber photometry or multichannel electrophysiological recordings in mice, we found that glutamatergic neurons of the paraventricular thalamus (PVT) exhibited high activities during wakefulness. Suppression of PVT neuronal activity caused a reduction in wakefulness, whereas activation of PVT neurons induced a transition from sleep to wakefulness and an acceleration of emergence from general anesthesia. Moreover, our findings indicate that the PVT-nucleus accumbens projections and hypocretin neurons in the lateral hypothalamus to PVT glutamatergic neurons' projections are the effector pathways for wakefulness control. These results demonstrate that the PVT is a key wakefulness-controlling nucleus in the thalamus.


Assuntos
Núcleos da Linha Média do Tálamo/fisiologia , Vigília/fisiologia , Animais , Eletrofisiologia/métodos , Feminino , Ácido Glutâmico , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Mutantes , Neurônios/fisiologia , Núcleo Accumbens/fisiologia , Optogenética , Orexinas/genética , Fotometria/métodos , Proteínas Proto-Oncogênicas c-fos/metabolismo
12.
Nat Neurosci ; 21(11): 1541-1550, 2018 11.
Artigo em Inglês | MEDLINE | ID: mdl-30361547

RESUMO

Urination (also called micturition) is thought to be regulated by a neural network that is distributed in both subcortical and cortical regions. Previously, urination-related neurons have been identified in subcortical structures such as the pontine micturition center (also known as Barrington's nucleus). However, the origin of the descending cortical pathway and how it interfaces with this subcortical circuit to permit voluntary initiation of urination remain elusive. Here we identified a small cluster of layer 5 neurons in the primary motor cortex whose activities tightly correlate with the onset of urination in freely behaving mice and increase dramatically during territorial marking. Optogenetically activating these neurons elicits contraction of the bladder and initiates urination, through their projections to the pontine micturition center, while silencing or ablating them impairs urination and causes retention of urine. Together these results reveal a novel cortical component upstream of the pontine micturition center that is critically involved in urination.


Assuntos
Córtex Motor/fisiologia , Neurônios/fisiologia , Ponte/fisiologia , Bexiga Urinária , Micção/fisiologia , Animais , Camundongos , Vias Neurais/fisiologia , Medula Espinal/fisiologia , Bexiga Urinária/fisiologia
13.
Cell Mol Life Sci ; 75(19): 3569-3592, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30046841

RESUMO

Multicopper oxidases (MCOs) are a pervasive family of enzymes that oxidize a wide range of phenolic and nonphenolic aromatic substrates, concomitantly with the reduction of dioxygen to water. MCOs are usually divided into two functional classes: metalloxidases and laccases. Given their broad substrate specificity and eco-friendliness (molecular oxygen from air as is used as the final electron acceptor and they only release water as byproduct), laccases are regarded as promising biological green tools for an array of applications. Among these laccases, those of bacterial origin have attracted research attention because of their notable advantages, including broad substrate spectrum, wide pH range, high thermostability, and tolerance to alkaline environments. This review aims to summarize the significant research efforts on the properties, mechanisms and structures, laccase-mediator systems, genetic engineering, immobilization, and biotechnological applications of the bacteria-source laccases and laccase-like enzymes, which principally include Bacillus laccases, actinomycetic laccases and some other species of bacterial laccases. In addition, these enzymes may offer tremendous potential for environmental and industrial applications.


Assuntos
Bactérias/enzimologia , Bioengenharia/tendências , Química Verde/tendências , Indústrias/tendências , Lacase/fisiologia , Animais , Bactérias/genética , Bioengenharia/métodos , Microbiologia Ambiental , Química Verde/métodos , Humanos , Indústrias/métodos , Invenções/tendências , Lacase/genética
14.
Neuron ; 99(1): 47-55.e4, 2018 07 11.
Artigo em Inglês | MEDLINE | ID: mdl-29909996

RESUMO

The ability to remember and to navigate to safe places is necessary for survival. Place navigation is known to involve medial entorhinal cortex (MEC)-hippocampal connections. However, learning-dependent changes in neuronal activity in the distinct circuits remain unknown. Here, by using optic fiber photometry in freely behaving mice, we discovered the experience-dependent induction of a persistent-task-associated (PTA) activity. This PTA activity critically depends on learned visual cues and builds up selectively in the MEC layer II-dentate gyrus, but not in the MEC layer III-CA1 pathway, and its optogenetic suppression disrupts navigation to the target location. The findings suggest that the visual system, the MEC layer II, and the dentate gyrus are essential hubs of a memory circuit for visually guided navigation.


Assuntos
Região CA1 Hipocampal/fisiologia , Giro Denteado/fisiologia , Córtex Entorrinal/fisiologia , Memória Espacial/fisiologia , Navegação Espacial , Animais , Sinais (Psicologia) , Hipocampo/fisiologia , Memória/fisiologia , Camundongos , Inibição Neural , Optogenética , Fotometria , Percepção Visual
15.
J Neuroinflammation ; 15(1): 136, 2018 May 04.
Artigo em Inglês | MEDLINE | ID: mdl-29728120

RESUMO

BACKGROUND: Interleukin-33 (IL-33) is increasingly being recognized as a key immunomodulatory cytokine in many neurological diseases. METHODS: In the present study, wild-type (WT) and IL-33-/- mice received intracerebroventricular (i.c.v.) injection of lipopolysaccharide (LPS) to induce neuroinflammation. Intravital microscopy was employed to examine leukocyte-endothelial interactions in the brain vasculature. The degree of neutrophil infiltration was determined by myeloperoxidase (MPO) staining. Real-time PCR and western blotting were used to detect endothelial activation. Enzyme-linked immunosorbent assay and quantitative PCR were conducted to detect pro-inflammatory cytokine levels in the brain. RESULTS: In IL-33-/- mice, neutrophil infiltration in the brain cortex and leukocyte-endothelial cell interactions in the cerebral microvessels were significantly decreased as compared to WT mice after LPS injection. In addition, IL-33-/- mice showed reduced activation of microglia and cerebral endothelial cells. In vitro results indicated that IL-33 directly activated cerebral endothelial cells and promoted pro-inflammatory cytokine production in LPS-stimulated microglia. CONCLUSIONS: Our study indicated that IL-33/ST2 signaling plays an important role in the activation of microglia and cerebral endothelial cells and, therefore, is essential in leukocyte recruitment in brain inflammation. The role of IL-33/ST2 in LPS induced neuroinflammation.


Assuntos
Encefalite/metabolismo , Células Endoteliais/metabolismo , Proteína 1 Semelhante a Receptor de Interleucina-1/biossíntese , Interleucina-33/biossíntese , Microglia/metabolismo , Animais , Animais Recém-Nascidos , Células Cultivadas , Encefalite/induzido quimicamente , Encefalite/imunologia , Células Endoteliais/imunologia , Imunidade Inata/fisiologia , Injeções Intraventriculares , Proteína 1 Semelhante a Receptor de Interleucina-1/imunologia , Interleucina-33/imunologia , Lipopolissacarídeos/toxicidade , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Microglia/imunologia , Infiltração de Neutrófilos/fisiologia
16.
Front Neural Circuits ; 12: 33, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29740289

RESUMO

In vivo two-photon Ca2+ imaging is a powerful tool for recording neuronal activities during perceptual tasks and has been increasingly applied to behaving animals for acute or chronic experiments. However, the auditory cortex is not easily accessible to imaging because of the abundant temporal muscles, arteries around the ears and their lateral locations. Here, we report a protocol for two-photon Ca2+ imaging in the auditory cortex of head-fixed behaving mice. By using a custom-made head fixation apparatus and a head-rotated fixation procedure, we achieved two-photon imaging and in combination with targeted cell-attached recordings of auditory cortical neurons in behaving mice. Using synthetic Ca2+ indicators, we recorded the Ca2+ transients at multiple scales, including neuronal populations, single neurons, dendrites and single spines, in auditory cortex during behavior. Furthermore, using genetically encoded Ca2+ indicators (GECIs), we monitored the neuronal dynamics over days throughout the process of associative learning. Therefore, we achieved two-photon functional imaging at multiple scales in auditory cortex of behaving mice, which extends the tool box for investigating the neural basis of audition-related behaviors.


Assuntos
Córtex Auditivo/fisiologia , Comportamento Animal/fisiologia , Dendritos/fisiologia , Espinhas Dendríticas/fisiologia , Animais , Masculino , Camundongos Endogâmicos C57BL , Neuroimagem/métodos , Neurônios/fisiologia
17.
J Sep Sci ; 41(10): 2130-2138, 2018 May.
Artigo em Inglês | MEDLINE | ID: mdl-29430840

RESUMO

Shenxiong glucose injection, a pharmaceutical preparation containing a water extract of the roots of Salvia miltiorrhizae and ligustrazine hydrochloride, is widely used in clinical to treat cardiovascular diseases in China. The chemical components of the water extract have been reported and the cardioprotective effects of the injection have been evaluated. However, the chemical constituents of the injection and their correlations with its pharmacological effects have not been established. In this study, 13 chemical constituents of the injection have been identified or characterized by ultra-high performance liquid chromatography with diode array detection and electrospray ionization quadrupole time-of-flight tandem mass spectrometry. Besides, the potentially active compounds of this preparation that directly act on cardiac cells have been screened by cell extraction and ultra high performance liquid chromatography targeted multiple reaction monitoring. As a result, eight potentially active compounds, danshensu (1), ligustrazine hydrochloride (4), salvianolic acid I/H (7), lithospermic acid (8), salvianolic acid D (9), rosmarinic acid (10), salvianolic acid B (12), and salvianolic acid C (13), were obtained and structurally characterized from the 11 target compounds used for screening. The liquid chromatography with quadrupole time-of-flight mass spectrometry and liquid chromatography with multiple reaction monitoring tandem mass spectrometry combination method has demonstrated its potency for the screening, detection, and structural identification of bioactive compounds in a complex matrix.


Assuntos
Cromatografia Líquida de Alta Pressão , Medicamentos de Ervas Chinesas/análise , Espectrometria de Massas em Tandem , Animais , Linhagem Celular , Concentração de Íons de Hidrogênio , Luz , Raízes de Plantas/química , Pirazinas/química , Ratos , Salvia miltiorrhiza/química , Espectrometria de Massas por Ionização por Electrospray , Água/química
18.
Sci Rep ; 8(1): 49, 2018 01 08.
Artigo em Inglês | MEDLINE | ID: mdl-29311673

RESUMO

In the primary auditory cortex (A1), neuronal ensembles are activated relative to anticipated sound events following rhythmic stimulation, but whether the echo responses of the neurons are related to their frequency selectivity remains unknown. Therefore, we used in vivo two-photon Ca2+ imaging to record the neuronal activities in the mouse A1 to elucidate the relationship between their echo responses and frequency selectivity. We confirmed the presence of echo responses in a subgroup of mouse Layer 2/3 A1 neurons following a train of rhythmic pure tone stimulation. After testing with a range of frequencies, we found that these echo responses occurred preferentially close to the best frequencies of the neurons. The local organization of the echo responses of the neurons was heterogeneous in the A1. Therefore, these results indicate that the observed echo responses of neurons within A1 are highly related to their frequency selectivity.


Assuntos
Córtex Auditivo/fisiologia , Percepção Auditiva , Estimulação Acústica , Animais , Mapeamento Encefálico , Cálcio/metabolismo , Fenômenos Eletrofisiológicos , Potenciais Evocados Auditivos , Masculino , Camundongos , Imagem Molecular/métodos , Neurônios/metabolismo , Tempo de Reação
19.
Int J Rheum Dis ; 21(2): 477-486, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29316328

RESUMO

AIM: To determine whether pirfenidone attenuates lung fibrosis by interfering with the hedgehog (Hh) signalling pathway in patients with systemic sclerosis-associated interstitial lung disease (SSc-ILD). METHODS: Twenty-five SSc-ILD patients (20 first visit, five who underwent pirfenidone treatment for 6 months) and 10 healthy controls were recruited. Lung tissues were obtained by open-chest surgery, and primary lung fibroblasts were isolated, cultured and stimulated with pirfenidone. The levels of the proteins glioma-associated oncogene 1 (GLI1), suppressor of fused (Sufu), α-smooth muscle actin, and fibronectin in lung tissues or fibroblasts were determined by Western blotting. The messenger RNA levels of GLI1, glioma-associated oncogene 2, protein patched homolog 1, and Sufu in lung tissues or fibroblasts were determined by quantitative reverse-transcription polymerase chain reaction. Meanwhile, the levels of phosphorylation glycogen synthase kinasep-3ß (pGSK-3ß), phosphorylation SMAD2 (pSMAD2), and phosphorylation c-Jun N-terminal kinase (pJNK) in fibroblasts were determined by Western blotting. RESULTS: Hh pathway activation was increased in the lung tissue of SSc-ILD patients and was decreased by pirfenidone, Sufu was upregulated in lung fibroblasts isolated from SSc-ILD patients after pirfenidone challenge, and pirfenidone inhibited the phosphorylation of GSK-3ß signalling. CONCLUSION: Pirfenidone has anti-fibrotic effects in SSc-ILD patients by interfering with both the Hh signalling pathway and the GSK-3ß signalling pathway via the regulation of Sufu expression. These results might promote its use in other Hh driven lung diseases such as idiopathic pulmonary fibrosis and especially the interstitial lung disease associated with connective tissue diseases.


Assuntos
Fibroblastos/efeitos dos fármacos , Proteínas Hedgehog/metabolismo , Doenças Pulmonares Intersticiais/tratamento farmacológico , Pulmão/efeitos dos fármacos , Fibrose Pulmonar/tratamento farmacológico , Piridonas/farmacologia , Escleroderma Sistêmico/tratamento farmacológico , Transdução de Sinais/efeitos dos fármacos , Actinas/metabolismo , Adulto , Estudos de Casos e Controles , Células Cultivadas , Feminino , Fibroblastos/metabolismo , Fibroblastos/patologia , Fibronectinas/metabolismo , Glicogênio Sintase Quinase 3 beta/metabolismo , Humanos , Pulmão/metabolismo , Pulmão/patologia , Doenças Pulmonares Intersticiais/diagnóstico , Doenças Pulmonares Intersticiais/metabolismo , MAP Quinase Quinase 4/metabolismo , Masculino , Pessoa de Meia-Idade , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Receptor Patched-1/genética , Receptor Patched-1/metabolismo , Fosforilação , Fibrose Pulmonar/diagnóstico , Fibrose Pulmonar/metabolismo , Proteínas Repressoras/genética , Proteínas Repressoras/metabolismo , Escleroderma Sistêmico/diagnóstico , Escleroderma Sistêmico/metabolismo , Proteína Smad2/metabolismo , Tomografia Computadorizada por Raios X , Proteína GLI1 em Dedos de Zinco/genética , Proteína GLI1 em Dedos de Zinco/metabolismo , Proteína Gli2 com Dedos de Zinco/genética , Proteína Gli2 com Dedos de Zinco/metabolismo
20.
Brain Struct Funct ; 223(1): 519-533, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-29124351

RESUMO

Two-photon Ca2+ imaging has become a popular approach for monitoring neuronal population activity with cellular or subcellular resolution in vivo. This approach allows for the recording of hundreds to thousands of neurons per animal and thus leads to a large amount of data to be processed. In particular, manually drawing regions of interest is the most time-consuming aspect of data analysis. However, the development of automated image analysis pipelines, which will be essential for dealing with the likely future deluge of imaging data, remains a major challenge. To address this issue, we developed NeuroSeg, an open-source MATLAB program that can facilitate the accurate and efficient segmentation of neurons in two-photon Ca2+ imaging data. We proposed an approach using a generalized Laplacian of Gaussian filter to detect cells and weighting-based segmentation to separate individual cells from the background. We tested this approach on an in vivo two-photon Ca2+ imaging dataset obtained from mouse cortical neurons with differently sized view fields. We show that this approach exhibits superior performance for cell detection and segmentation compared with the existing published tools. In addition, we integrated the previously reported, activity-based segmentation into our approach and found that this combined method was even more promising. The NeuroSeg software, including source code and graphical user interface, is freely available and will be a useful tool for in vivo brain activity mapping.


Assuntos
Cálcio/metabolismo , Processamento Eletrônico de Dados/métodos , Neurônios/citologia , Neurônios/fisiologia , Algoritmos , Animais , Mapeamento Encefálico , Tamanho Celular , Processamento Eletrônico de Dados/instrumentação , Processamento de Imagem Assistida por Computador , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Distribuição Normal , Óptica e Fotônica , Software
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA