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Here, lignin and nano-clay were used to prepare novel composite adsorbents by one-step carbonization without adding activators for radioactive iodine capture. Specially, 1D nano-clay such as halloysite (Hal), palygorskite (Pal) and sepiolite (Sep) were selected as skeleton components, respectively, enzymatic hydrolysis lignin (EHL) as carbon source, lignin based porous carbon/nano-clay composites (ELC-X) were prepared through ultrasonic impregnation, freeze drying, and carbonization. Characterization results indicated lignin based porous carbon (ELC) well coated on the surface of nano-clay, and made its surface areas increase to 252 m2/g. These composites appeared the micro-mesoporous hierarchical structure, considerable N doping and good chemical stability. Results of adsorption experiments showed that the introduction of ELC could well promote iodine vapor uptake of nano-clay, and up to 435.0 mg/g. Meanwhile, the synergistic effect between lignin based carbon and nano-clay was very significant for the adsorption of iodine/n-hexane and iodine ions, their capacity were far exceed those of a single material, respectively. The relevant adsorption kinetic and thermodynamics, and mechanism of ELC-X composites were clarified. This work provided a class of low-cost and environmentally friendly adsorbents for radioactive iodine capture, and opened up ideas for the comprehensive utilization of waste lignin and natural clay minerals.
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Carbono , Argila , Iodo , Lignina , Lignina/química , Argila/química , Carbono/química , Adsorção , Porosidade , Iodo/química , Cinética , Nanocompostos/química , TermodinâmicaRESUMO
Introduction: The capacity to speak English for academic purposes is a pivotal facet of language education and assessment. Despite the substantial research approving the significant role of teachers in L2 learning, it remains unclear how exactly teachers' support for students' learning autonomy influences EFL learners' academic English speaking performance. Methods: To address this primary concern, this study drew ground from the Self-System Model of Motivational Development (SSMMD) and adopted a mixed-method approach to examine teacher autonomy support's direct and indirect effects on Chinese EFL undergraduates' academic English speaking performance through the mediation of basic psychological needs and classroom engagement. 247 first-year university students participating in academic English speaking courses were recruited in this study. Results and discussion: The quantitative results of the questionnaire indicated that students' perceived teacher support for autonomy directly predicted English speaking performance, and it also indirectly influenced students' speaking performance via the mediation of classroom engagement and basic psychological needs. Students' responses in the semi-structured interview further verified the positive effect of teacher autonomy support on academic English speaking development in the classroom. Pedagogical implications were also discussed based on the findings.
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In this study, the lignin was pre-modified using small-molecule nitrogen-containing compounds, and then the nitrogen-doped lignin-based carbon nanotubes (L-NCNTs) were fabricated by pyrolysis using the modified lignin as raw materials. The obtained L-NCNTs were multi-walled carbon nanotubes with diameters between 10 and 80 nm. The modification of lignin had an important effect on the nitrogen morphology of L-NCNTs, and promoted the high selectivity of pyridine-N in the L-NCNTs. Defects and pyridinic-N structure were conducive to boosting photothermal properties of the L-NCNTs. The photothermal conversion efficiency of the L-NCNTs after 808 nm laser irradiation for 5 min reached 58.8 %. The L-NCNTs can be used as photothermal agents in drug delivery system to achieve mild photothermal therapy, and it is basically non-toxic to normal cells, indicating good biocompatibility. This work provides new ideas for development of lignin-based high value-added products from biomass.
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Nanotubos de Carbono , Nanotubos de Carbono/química , Nitrogênio/química , Lignina , Terapia Fototérmica , Compostos de NitrogênioRESUMO
BACKGROUND: The postoperative delirium is a common yet serious complication in elderly patients with hip fracture. We aimed to evaluate the potential risk factors of delirium in patients with hip fracture, to provide reliable evidence to the clinical management of hip fracture. METHODS: This study was a retrospective design. Elderly patients who underwent hip fracture surgery in our hospital from June 1, 2019 to December 30, 2020 were selected. The characteristics and treatment data of delirium and no delirium patients were collected and compared. Multivariate logistic regression analysis was performed to analyze the influencing factors affecting postoperative delirium in elderly patients with hip fracture. RESULTS: A total of 245 patients with hip fracture were included, the incidence of postoperative delirium in patients with hip fracture was 13.06%. There were significant differences in the age, BMI, history of delirium, estimated blood loss and duration of surgery (all p < 0.05). There were significant differences in the albumin and TSH between delirium and no delirium group (all p < 0.05), Logistics analyses indicated that age ≥ 75 years (OR 3.112, 95% CI 1.527-5.742), BMI ≥ 24 kg/m2 (OR 2.127, 95% CI 1.144-3.598), history of delirium (OR 1.754, 95% CI 1.173-2.347), estimated blood loss ≥ 400 mL (OR 1.698, 95% CI 1.427-1.946), duration of surgery ≥ 120 min (OR 2.138, 95% CI 1.126-3.085), preoperative albumin ≤ 40 g/L (OR 1.845, 95% CI 1.102-2.835) and TSH ≤ 2 mU/L (OR 2.226, 95% CI 1.329-4.011) were the independent risk factors of postoperative delirium in patients with hip fracture(all p < 0.05). CONCLUSIONS: Postoperative delirium is very common in elderly patients with hip fracture, and it is associated with many risk factors, clinical preventions targeted on those risk factors are needed to reduce the postoperative delirium.
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Delírio/epidemiologia , Fraturas do Quadril/cirurgia , Ossos Pélvicos/lesões , Complicações Pós-Operatórias/epidemiologia , Medição de Risco/métodos , Idoso , China/epidemiologia , Delírio/etiologia , Feminino , Seguimentos , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Complicações Pós-Operatórias/etiologia , Estudos Retrospectivos , Fatores de RiscoRESUMO
The objective of this study is to determine the feasibility and explore criteria for patient selection for three methods of LESS myomectomy: conventional (C-LESS), robotic-assisted (RA-LESS), and hand-assisted (HA-LESS). This was a retrospective case review of 72 patients with uterine myomas, conducted in a large academic tertiary care hospital between March 1, 2015, and November 7, 2018. LESS myomectomy via conventional, robotic, and hand-assisted routes. 43 patients underwent C-LESS, 15 underwent RA-LESS, and 14 underwent HA-LESS, with no conversions to open abdominal myomectomy. The operative outcomes were compared across the three approaches. The HA-LESS group had the largest mean number (HA: 6.9; C: 3.7; RA: 2.9, P=0.001), diameter (HA: 11.3 cm; C: 9.3 cm; RA: 7.1 cm, P=0.035), and weight (HA: 850.1 g; C: 320.7 g; RA: 181.1 g, P=0.003) of myomas removed per patient. The use of this method was also found to have a direct correlation with estimated preoperative uterine size (HA: 20.1 weeks; C: 16.2 weeks; RA: 12.0 weeks, P=0.001. Operative time and postoperative stay were found to be not statistically different across groups. We conclude that all three types of LESS myomectomy are feasible with comparable surgical outcomes. Most importantly, our findings indicate that hand assistance can be combined with C-LESS myomectomy for large or multi-fibroid uterus without compromising operating time or patient recovery. Notably, we found that uterine size could be a useful tool for the determination of the surgical approach.
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Laparoscopia , Procedimentos Cirúrgicos Robóticos , Miomectomia Uterina , Neoplasias Uterinas , Feminino , Humanos , Duração da Cirurgia , Estudos Retrospectivos , Procedimentos Cirúrgicos Robóticos/métodos , Neoplasias Uterinas/cirurgiaRESUMO
A pH-stimuli amphiphilic lignin-based copolymer was prepared, and it could self-assemble to form spherical nanomicelles with the addition of "switching" water. The morphology, structure, and physical properties of micelles were characterized with transmission electron microscopy (TEM), nuclear magnetic resonance (NMR), Fourier transform infrared spectroscopy (FTIR), gel permeation chromatography (GPC), particle-size analysis, and zeta-potential measurement. In vitro drug release exemplified that the micelles were pH-sensitive, retaining more than 84.36% ibuprofen (IBU) in simulated gastric fluid (pH 1.5) and presenting a smooth release of 81.81% IBU in simulated intestinal fluid (pH 7.4) within 72 h. Cell culture studies showed that the nanomicelles were biocompatible and boosted the proliferation of human bone marrow stromal cells hBMSC and mouse embryonic fibroblast cells NIH-3T3. Interestingly, the nanomicelles inhibited the survival of human colon cancer cells HT-29 with a final survival rate of only 5.34%. Therefore, this work suggests a novel strategy to synthesize intelligent lignin-based nanomicelles that show a great potential as oral drug carriers.
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Portadores de Fármacos/química , Sistemas de Liberação de Medicamentos/métodos , Ibuprofeno/química , Lignina/química , Animais , Linhagem Celular , Sistemas de Liberação de Medicamentos/instrumentação , Liberação Controlada de Fármacos , Humanos , Concentração de Íons de Hidrogênio , Ibuprofeno/farmacologia , Camundongos , Micelas , Tamanho da PartículaRESUMO
Cellulose nanocrystal (CNC)/poly(lactic acid) (PLA) in situ nanocomposite scaffolds were fabricated by in situ polymerization of lactic acid and CNC which was directly utilized as aqueous suspension, followed by a process of thermally induced phase separation. The CNC/PLA in situ nanocomposite porous scaffolds were characterized by mechanical test, protein adsorption, hemolysis test, in vitro degradation measurement, TEM, FTIR, SEM and WAXD. Compared to the PLA scaffold, the CNC/PLA in situ nanocomposite scaffolds showed a greatly increased compression modulus, an improved hemocompatibility and protein adsorption capacity. The inclusion of CNCs boosted the in vitro degradation of the in situ nanocomposite porous scaffolds and facilitated the deposition of Ca2+, CO32-, PO43- ions in simulated body fluid. Furthermore, cell cultures were carried out on the CNC/PLA in situ nanocomposite porous scaffolds. In comparison with the PLA scaffold, the in situ nanocomposite scaffolds improved cell attachment and enhanced cell proliferation, denoting low cytotoxicity and good cytocompatibility. It can therefore be concluded that such scaffolds with excellent mechanical property, biocompatibility, biomineralization capacity and bioactivity hold great potential for bone tissue engineering.
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Celulose/química , Nanocompostos/química , Nanopartículas/química , Poliésteres/química , Engenharia Tecidual , Alicerces Teciduais/química , Linhagem Celular Tumoral , Sobrevivência Celular , Hemólise , Humanos , Peso Molecular , Nanocompostos/ultraestrutura , Nanopartículas/ultraestrutura , Porosidade , Proteínas/química , Análise EspectralRESUMO
Hyperbranched polymers have attracted wide research attention owing to their unique topological structure, physicochemical properties and great potential for applications such asadditives, drug delivery, catalysts and nanotechnology. Among these, the polyamidoamine(PAMAM) dendrimers are some of the most important dendrimers. However, the synthesis and biomedical applications of fluorescent PAMAM dendrimers have received only limited attention. In this work, we present a rather effective and convenient approach for synthesis of fluorescent PAMAM dendrimers with aggregation-induced emission (AIE) properties through a one-pot catalyst-free Mannich reaction under rather mild experimental conditions (e.g., low reaction temperature, air atmosphere in the presence of water). The obtained AIE-active amphiphiles (PhE-PAD) could self-assemble into fluorescent organic nanoparticles (FONs). The obtained AIE-active FONs (PhE-PAD FONs) were fully characterized, and their successful construction was confirmed by 1H NMR spectroscopy, FT-IR spectroscopy and transmission electron microscopy. Fluorescence and UV-Visible absorption spectroscopy results demonstrated that the final PhE-PAD FONs showed strong yellow fluorescence, desirable photostability and good water dispersity. The cell viability evaluation and confocal laser scanning microscope imaging results suggested that PhE-PAD FONs possessed low cytotoxicity and excellent biocompatibility. Taken together, these results demonstrate that we have developed a facile and efficient strategy for the fabrication of AIE-active FONs, which possess many desirable features for biomedical applications.
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Dendrímeros/síntese química , Corantes Fluorescentes/síntese química , Imagem Óptica/métodos , Animais , Catálise , Linhagem Celular , Sobrevivência Celular , Luz , Teste de Materiais , Camundongos , Tamanho da Partícula , Espectrometria de Fluorescência , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
In this paper, an implicit finite difference scheme with the shifted Grünwald formula, which is unconditionally stable, is used to discretize the fractional diffusion equations with constant diffusion coefficients. The coefficient matrix possesses the Toeplitz structure and the fast Toeplitz matrix-vector product can be utilized to reduce the computational complexity from [Formula: see text] to [Formula: see text], where N is the number of grid points. Two preconditioned iterative methods, named bi-conjugate gradient method for Toeplitz matrix and bi-conjugate residual method for Toeplitz matrix, are proposed to solve the relevant discretized systems. Finally, numerical experiments are reported to show the effectiveness of our preconditioners.
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Cryo-imaging techniques have been widely used to measure the metabolic state of tissues by capturing reduced nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD) autofluorescence. However, NADH and FAD fluorescence is sensitive to changes in temperature, which may result in unreliable redox ratio calculations. Here, the relationship between the measured redox ratio and sample surface temperature was analyzed using a standard phantom solution and biological tissues. The results indicated that a temperature < - 100°C was a suitable cryo-imaging temperature window in which redox ratio measuring was immune to temperature fluctuations. These results may serve as a reference for designing and optimizing redox cryo-imaging experiments for quantitatively mapping the metabolic state of biological samples.
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Química Encefálica , Flavina-Adenina Dinucleotídeo/química , Secções Congeladas/instrumentação , Imagem Molecular/instrumentação , NAD/química , Espectrometria de Fluorescência/instrumentação , Termografia/instrumentação , Animais , Desenho de Equipamento , Análise de Falha de Equipamento , Congelamento , Aumento da Imagem/instrumentação , Aumento da Imagem/métodos , Iluminação , Oxirredução , Ratos , Reprodutibilidade dos Testes , Semicondutores , Sensibilidade e EspecificidadeRESUMO
Understanding the biological mechanism and identifying biomarkers of hemorrhagic shock is important for diagnosis and treatment. We aim to use optical imaging to study how the cerebral blood circulation and metabolism change during the progression of severe hemorrhagic shock, especially the decompensatory stage. We used a multi-parameter (blood pressure (BP), cerebral blood flow (CBF), functional vascular density (FVD), blood oxygenation and mitochondrial NADH signal) cerebral cortex optical imaging system to observe brain hemodynamic change and metabolic alteration of rats in vivo for 4 h. Cerebral circulation and mitochondrial metabolism could be well preserved in the compensatory stage but impaired during the decompensatory stage. The changes of brain hemodynamics and metabolism may provide sensitive indicators for various shock stages including the transition from compensatory stage to decompensatory stage. Our novel imaging observations of hemodynamic and metabolic signals in vivo indicated that the rat brains under hemorrhagic shock suffered irreversible damage which could not be compensated by the autoregulation mechanism, probably due to injured mitochondria.
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Hemodinâmica , Choque Hemorrágico/fisiopatologia , Humanos , Índice de Gravidade de Doença , Choque Hemorrágico/metabolismoRESUMO
The topology of the cerebral vasculature, which is the energy transport corridor of the brain, can be used to study cerebral circulatory pathways. Limited by the restrictions of the vascular markers and imaging methods, studies on cerebral vascular structure now mainly focus on either observation of the macro vessels in a whole brain or imaging of the micro vessels in a small region. Simultaneous vascular studies of arteries, veins and capillaries have not been achieved in the whole brain of mammals. Here, we have combined the improved gelatin-Indian ink vessel perfusion process with Micro-Optical Sectioning Tomography for imaging the vessel network of an entire mouse brain. With 17 days of work, an integral dataset for the entire cerebral vessels was acquired. The voxel resolution is 0.35×0.4×2.0 µm(3) for the whole brain. Besides the observations of fine and complex vascular networks in the reconstructed slices and entire brain views, a representative continuous vascular tracking has been demonstrated in the deep thalamus. This study provided an effective method for studying the entire macro and micro vascular networks of mouse brain simultaneously.
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Encéfalo/irrigação sanguínea , Encéfalo/fisiologia , Circulação Cerebrovascular/fisiologia , Microvasos/fisiologia , Animais , Encéfalo/metabolismo , Gelatina/metabolismo , Tinta , Camundongos , Perfusão/métodos , Tomografia/métodosRESUMO
RATIONALE AND OBJECTIVES: Our long-term goals are to identify imaging biomarkers for hemorrhagic shock and to understand how the preservation of cerebral microcirculation works. We also seek to understand how the damage occurs to the cerebral hemodynamics and the mitochondrial metabolism during severe hemorrhagic shock. MATERIALS AND METHODS: We used a multimodal cerebral cortex optical imaging system to obtain 4-hour observations of cerebral hemodynamic and metabolic alterations in exposed rat cortexes during severe hemorrhagic shock. We monitored the mean arterial pressure, heart rate, cerebral blood flow (CBF), functional vascular density (FVD), vascular perfusion and diameter, blood oxygenation, and mitochondrial reduced nicotinamide adenine dinucleotide (NADH) signals. RESULTS: During the rapid bleeding and compensatory stage, cerebral parenchymal circulation was protected by inhibiting the perfusion of dural vessels. During the compensatory stage, although the brain parenchymal CBF and FVD decreased rapidly, the NADH signal did not show a significant increase. During the decompensatory stage, FVD and CBF maintained the same low level and the NADH signal remained unchanged. However, the NADH signal showed a significant increase after the rapid blood infusion. FVD and CBF rebounded to the baseline after the resuscitation and then declined again. CONCLUSIONS: We present for the first time simultaneous imaging of cerebral hemodynamics and NADH signals in vivo during the process of hemorrhagic shock. This novel multimodal method demonstrated clearly that severe hemorrhagic shock imparts irreversible tissue damage that is not compensated by the autoregulatory mechanism. Hemodynamic and metabolic signatures including CBF, FVD, and NADH may be further developed to provide sensitive biomarkers for stage transitions in hemorrhagic shock.
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Encéfalo/metabolismo , Córtex Cerebral/fisiopatologia , Circulação Cerebrovascular , Imagem Molecular/métodos , NAD/metabolismo , Oxigênio/sangue , Choque Hemorrágico/fisiopatologia , Animais , Velocidade do Fluxo Sanguíneo , Masculino , Imagem Multimodal/métodos , Ratos , Ratos WistarRESUMO
The goal of biological samples' cryofixation is to trap a metabolic state as it exists in vivo by rapidly stopping internal reactions. However, obtaining perfect quality of cryofixation for large and high hypermetabolism organ/tissue (such as brain, heart) remains a challenge. The aim of this study was to develop and display a comprehensive and direct method to evaluate cryofixation's process and quality. Here, we adopt a delicate combination of homemade cryo-imaging system with a rat cardiac arrest model that can control cryofixation time optionally. we successfully evaluate the cryofixation time-related nicotinamide adenine dinucleotide (NADH) fluorescence pattern of several coronal sections in rat's brain that suffered from directional funnel cryofixation procedure. Through quantitative analysis of the distribution map of NADH fluorescence, we could obtain a relationship between cryofixation time and well cryofixation volume and then could deduce the cryofixation rates and quality at different time points. Our results also demonstrated that dissection of the temporal muscle of rat could significantly optimize the classical direct funnel cryofixation protocol.
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Encéfalo/metabolismo , Criopreservação/métodos , NAD/metabolismo , Espectrometria de Fluorescência/métodos , Animais , Fluorescência , Parada Cardíaca/metabolismo , Masculino , Ratos , Ratos WistarRESUMO
Cortical spreading depression (CSD) is a self-propagating wave of cellular depolarization that plays an important role in the development of cerebral pathology following ischemia or trauma. Optical intrinsic signal (OIS) imaging has been widely used to investigate CSD. Sources of OIS are complex and related to the changes in brain tissue absorption and scattering. The absorbing chromophores may include oxy-hemoglobin, deoxy-hemoglobin, cytochromes, flavin adenine dinucleotide (FAD) and nicotinamide adenine dinucleotide (NADH). Considering only one or part of these elements in studies involving OIS may cause inaccurate results. Thus, we simultaneously calculated changes in HbO, HbR, FAD, cytochrome c, cytochrome aa3 and light scattering during CSD by applying multi-spectral OIS imaging at 450, 470, 500, 530, 550, 570, 600, 630, and 650 nm in the rat brain. We also showed that the hemodynamic changes during CSD may not be correctly estimated if the scattering and other chromophores such as FAD, cytochrome c and cytochrome aa3, are not included in the fitting model of multi-wavelength data analysis. As shown in our results, if considering the changes in scattering and other chromophores in data fitting model, deoxy-hemoglobin (HbR) showed a triphasic change while only a monophasic decrease in HbR will be resolved without considering changes in scattering and other chromophores as reported in previous studies. Moreover, our results showed that changes in cytochrome c was tightly related to OIS at 550 nm, cytochrome aa3 was closely related to OIS at 450, 600 and 650 nm, and FAD was closely related to OIS at 450 and 470 nm during CSD. It indicates that if the contribution by these related chromophores is not considered, using OIS at these wavelengths to determine the hemoglobin changes during CSD may lead to inaccurate results.
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Córtex Cerebral/fisiologia , Depressão Alastrante da Atividade Elétrica Cortical/fisiologia , Hemodinâmica/fisiologia , Mitocôndrias/metabolismo , Imagem Óptica/métodos , Animais , Masculino , Ratos , Ratos WistarRESUMO
Cerebral blood flow (CBF) is critical for the maintenance of cerebral function by guaranteed constant oxygen and glucose supply to brain. Collateral channels (CCs) are recruited to provide alternatives to CBF to ischemic regions once the primary vessel is occluded during ischemic stroke. However, the knowledge of the relationship between dynamic evolution of collateral flow and the distribution of regional blood flow remains limited. In this study, laser speckle imaging was used to assess dynamic changes of CCs and regional blood flow in a rat cortex with permanent middle cerebral artery occlusion (MCAo). We found that CCs immediately provided blood flow to ischemic territories after MCAo. More importantly, there were three kinds of dynamic changes of CCs during acute stroke: persistent CC, impermanent CC, and transient CC, respectively, related to different distributions of regional blood flow. Although there was the possible occurrence of peri-infarct depolarization (PID) during ischemia, there was no obvious significance about the onset time and duration of CCs between rats with and without PID. These results suggest that the initial arising of CCs does not ensure their persistence, and that collateral flow could be varied with distribution of regional blood flow in acute ischemic stroke, which may facilitate the understanding of collateral recruitment and promote the development of collateral therapeutics in the future.
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Isquemia Encefálica/fisiopatologia , Córtex Cerebral/irrigação sanguínea , Córtex Cerebral/fisiopatologia , Circulação Cerebrovascular , Circulação Colateral , Fluxometria por Laser-Doppler/métodos , Fluxo Sanguíneo Regional , Animais , Velocidade do Fluxo Sanguíneo , Masculino , Ratos , Ratos Sprague-Dawley , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
Laser speckle spatial contrast analysis (LSSCA) is superior to laser speckle temporal contrast analysis (LSTCA) in monitoring the fast change in blood flow due to its advantage of high temporal resolution. However, the application of LSSCA which is based on spatial statistics may be limited when there is nonuniform intensity distribution such as fiber-transmitting laser speckle imaging. In this study, we present a normalized laser speckle spatial contrast analysis (nLSSCA) to correct the detrimental effects of nonuniform intensity distribution on the spatial statistics. Through numerical simulation and phantom experiments, it is found that just ten frames of dynamic laser speckle images are sufficient for nLSSCA to achieve effective correction. Furthermore, nLSSCA has higher temporal resolution than LSTCA to respond the change in velocity. LSSCA, LSTCA and nLSSCA are all applied in the fiber-transmitting laser speckle imaging system to analyze the change of cortical blood flow (CBF) during cortical spreading depression (CSD) in rat cortex respectively, and the results suggest that nLSSCA can examine the change of CBF more accurately. For these advantages, nLSSCA could be a potential tool for fiber-transmitting/endoscopic laser speckle imaging.
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Artérias/fisiologia , Artefatos , Velocidade do Fluxo Sanguíneo/fisiologia , Tecnologia de Fibra Óptica/instrumentação , Aumento da Imagem/instrumentação , Fluxometria por Laser-Doppler/instrumentação , Humanos , Imagens de Fantasmas , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
Automatic separation of arteries and veins in optical cerebral cortex images is important in clinical practice and preclinical study. In this paper, a simple but effective automatic artery-vein separation method which utilizes single-wavelength coherent illumination is presented. This method is based on the relative temporal minimum reflectance analysis of laser speckle images. The validation is demonstrated with both theoretic simulations and experimental results applied to the rat cortex. Moreover, this method can be combined with laser speckle contrast analysis so that the artery-vein separation and blood flow imaging can be simultaneously obtained using the same raw laser speckle images data to enable more accurate analysis of changes of cerebral blood flow within different tissue compartments during functional activation, disease dynamic, and neurosurgery, which may broaden the applications of laser speckle imaging in biology and medicine.
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Artérias/fisiologia , Automação , Circulação Cerebrovascular/fisiologia , Imageamento Tridimensional/métodos , Lasers , Veias/fisiologia , Animais , Artérias/anatomia & histologia , Velocidade do Fluxo Sanguíneo/fisiologia , Ratos , Fatores de Tempo , Veias/anatomia & histologiaRESUMO
Cortical spreading depression (CSD) plays an important role in trauma, migraine and ischemia. CSD could induce pronounced hemodynamic changes and the disturbance of pH homeostasis which has been postulated to contribute to cell death following ischemia. In this study, we described a fluorescence-corrected multimodal optical imaging system to simultaneously monitor CSD associated intracellular pH (pH(i)) changes and hemodynamic response including hemoglobin concentrations and cerebral blood flow (CBF). CSD was elicited by application of KCl on rat cortex and direct current (DC) potential was recorded as a typical characteristic of CSD. The pH(i) shift was mapped by neutral red (NR) fluorescence which was excited at 516-556 nm and emitted at 625 nm. The changes in hemoglobin concentrations were determined by dual-wavelength optical intrinsic signal imaging (OISI) at 550 nm and 625 nm. Integration of fluorescence imaging and dual-wavelength OISI was achieved by a time-sharing camera equipped with a liquid crystal tunable filter (LCTF). CBF was visualized by laser speckle contrast imaging (LSCI) through a separate camera. Besides, based on the dual-wavelength optical intrinsic signals (OISs) obtained from our system, NR fluorescence was corrected according to our method of fluorescence correction. We found that a transient intracellular acidification followed by a small alkalization occurred during CSD. After CSD, there was a prolonged intracellular acidification and the recovery of pH(i) from CSD took much longer time than those of hemodynamic response. Our results suggested that the new multimodal optical imaging system had the potential to advance our knowledge of CSD and might work as a useful tool to exploit neurovascular coupling under physiological and pathological conditions.