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δ-MnO2 is an important component of environmental minerals and is among the strongest sorbents and oxidants. The crystalline morphology of δ-MnO2 is one of the key factors affecting its reactivity. In this work, δ-MnO2 was initially synthesized and placed in an acidic environment to react with Mn2+ and undergo a crystalline transformation. During the transformation of crystalline δ-MnO2, kinetic sampling was conducted, followed by analyses of the structures and morphologies of the samples. The results showed that at pH 2.5 and 4, δ-MnO2 nanoflakes spontaneously self-assembled into nanoribbons via edge-to-edge assembly in the initial stage. Subsequently, these nanoribbons attached to each other to form primary nanorods through a face-to-face assembly along the c-axis. These primary nanorods then assembled along the (001) planes and lateral surfaces, achieving further growth and thickening. Since a lower pH is more favorable for the formation of vacancies in δ-MnO2, δ-MnO2 can rapidly adsorb Mn2+ directly onto the vacancies to form tunnel walls. At the same time, the rapid formation of the tunnel walls leads to a quick establishment of hydrogen bonding between adjacent nanoribbons, enabling the assembly of these nanoribbons into primary nanorods. Therefore, in a solution with the same concentration of Mn2+, the structure transformation and morphology evolution of δ-MnO2 to α-MnO2 occur faster at pH 2.5 than at pH 4. These findings provide insights into the mechanism for crystal growth from layer-based to tunnel-based nanorods and methods for efficient and controlled syntheses of nanomaterials.
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Advances in microfluidic device miniaturization and system integration contribute to the development of portable, handheld, and smartphone-compatible devices. These advancements in diagnostics have the potential to revolutionize the approach to detect and respond to future pandemics. Accordingly, herein, recent advances in point-of-care testing (POCT) of coronavirus disease 2019 (COVID-19) using various microdevices, including lateral flow assay strips, vertical flow assay strips, microfluidic channels, and paper-based microfluidic devices, are reviewed. However, visual determination of the diagnostic results using only microdevices leads to many false-negative results due to the limited detection sensitivities of these devices. Several POCT systems comprising microdevices integrated with portable optical readers have been developed to address this issue. Since the outbreak of COVID-19, effective POCT strategies for COVID-19 based on optical detection methods have been established. They can be categorized into fluorescence, surface-enhanced Raman scattering, surface plasmon resonance spectroscopy, and wearable sensing. We introduced next-generation pandemic sensing methods incorporating artificial intelligence that can be used to meet global health needs in the future. Additionally, we have discussed appropriate responses of various testing devices to emerging infectious diseases and prospective preventive measures for the post-pandemic era. We believe that this review will be helpful for preparing for future infectious disease outbreaks.
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COVID-19 , Humanos , COVID-19/diagnóstico , Inteligência Artificial , Estudos Prospectivos , Testes Imediatos , Sistemas Automatizados de Assistência Junto ao Leito , Teste para COVID-19RESUMO
Ultrasensitive detection of specific biomarkers in clinical serum is helpful for early diagnosis of cervical cancer. In this paper, a surface-enhanced Raman scattering (SERS)-based immunoassay was developed for the simultaneous determination of squamous cell carcinoma antigen (SCCA) and osteopontin (OPN) in cervical cancer serum. Au-Ag nanoshuttles (Au-AgNSs) as SERS tags and hydrophobic filter paper-based Au nanoflowers (AuNFs) as capture substrate were constructed into a sandwich structure which served as an ultrasensitive SERS-based immunoassay platform. Finite difference time domain simulation confirmed that the electromagnetic field coupled between the AuNFs had a prominent SERS signal enhancement effect, which improved the detection sensitivity. SERS mapping showed that hexadecenyl succinic anhydride hydrophobic treatment could prevent the analyte from being quickly absorbed by the filter paper and increase the retention time to be more evenly distributed on the filter paper substrate. The immunoassay platform was verified to have good selectivity and reproducibility. With this method, the detection limits of SCCA and OPN in human serum were as low as 8.628 pg/mL and 4.388 pg/mL, respectively. Finally, in order to verify the feasibility of its clinical application, the serum samples of healthy subjects; cervical intraepithelial neoplasia I (CINI), CINII, and CINIII; and cervical cancer patients were analyzed, and the reliability of the results was confirmed by enzyme-linked immunosorbent assay experiments. The constructed SERS-based immunoassay platform could be used as a clinical tool for early screening of cancers in the future.
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Biomarcadores Tumorais/sangue , Ouro/química , Nanopartículas Metálicas/química , Papel , Prata/química , Análise Espectral Raman/métodos , Neoplasias do Colo do Útero/diagnóstico , Adulto , Feminino , Humanos , Interações Hidrofóbicas e Hidrofílicas , Imunoensaio/métodos , Limite de Detecção , Neoplasias do Colo do Útero/sangue , Adulto JovemRESUMO
In vivo drug monitoring is crucial for evaluating the effectiveness and safety of drug treatment. Blood sampling and analysis is the current gold standard but needs professional skills and cannot meet the requirements of point-of-care testing. Dermal interstitial fluid (ISF) showed great potential to replace blood for in vivo drug monitoring; however, the detection was challenging, and the drug distribution behavior in ISF was still unclear until now. In this study, we proposed surface-enhanced Raman spectroscopy (SERS) microneedles (MNs) for the painless and real-time analysis of drugs in ISF after intravenous injection. Using methylene blue (MB) and mitoxantrone (MTO) as model drugs, the innovative core-satellite structured Au@Ag SERS substrate, hydrogel coating over the MNs, rendered sensitive and quantitative drug detection in ISF of mice within 10 min. Based on this technique, the pharmacokinetics of the two drugs in ISF was investigated and compared with those in blood, where the drugs were analyzed via liquid chromatography-mass spectrometry. It was found that the MB concentration in ISF and blood was comparable, whereas the concentration of MTO in ISF was 2-3 orders of magnitude lower than in blood. This work proposed an efficient tool for ISF drug monitoring. More importantly, it experimentally proved that the penetration ratio of blood to ISF was drug-dependent, providing insightful information into the potential of ISF as a blood alternative for in vivo drug detection.
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Monitoramento de Medicamentos , Líquido Extracelular , Hidrogéis , Azul de Metileno , Agulhas , Análise Espectral Raman , Animais , Análise Espectral Raman/métodos , Líquido Extracelular/química , Azul de Metileno/química , Camundongos , Hidrogéis/química , Monitoramento de Medicamentos/métodos , Monitoramento de Medicamentos/instrumentação , Prata/química , Mitoxantrona/sangue , Mitoxantrona/análise , Mitoxantrona/farmacocinética , Ouro/química , Pele/metabolismo , Pele/químicaRESUMO
Polystyrene nanoplastics are a novel class of pollutants. They are easily absorbed by living organisms, and their potential toxicity has raised concerns. However, the impact of polystyrene nanoplastics on auditory organs remains unknown. Here, our results showed that polystyrene nanoplastics entered the cochlea of mice, HEI-OC1 cells, and lateral line hair cells of zebrafish, causing cellular injury and increasing apoptosis. Additionally, we found that exposure to polystyrene nanoplastics resulted in a significant elevation in the auditory brainstem response thresholds, a loss of auditory sensory hair cells, stereocilia degeneration and a decrease in expression of Claudin-5 and Occludin proteins at the blood-lymphatic barrier in mice. We also observed a significant decrease in the acoustic alarm response of zebrafish after exposure to polystyrene nanoplastics. Mechanistic analysis revealed that polystyrene nanoplastics induced up-regulation of the Nrf2/HO-1 pathway, increased levels of malondialdehyde, and decreased superoxide dismutase and catalase levels in cochlea and HEI-OC1 cells. Furthermore, we observed that the expression of ferroptosis-related indicators GPX4 and SLC7A11 decreased as well as increased expression of ACLS4 in cochlea and HEI-OC1 cells. This study also revealed that polystyrene nanoplastics exposure led to increased expression of the inflammatory factors TNF-α, IL-1ß and COX2 in cochlea and HEI-OC1 cells. Further research found that the cell apoptosis, ferroptosis and inflammatory reactions induced by polystyrene nanoplastics in HEI-OC1 cells was reversed through the pretreatment with N-acetylcysteine, a reactive oxygen species inhibitor. Overall, our study first discovered and systematically revealed the ototoxicity of polystyrene nanoplastics and its underlying mechanism.
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Introduction: Rice (Oryza sativa L.) is a pivotal cereal crop worldwide. It relies heavily on the presence of iron plaque on its root surfaces for optimal growth and enhanced stress resistance across diverse environmental conditions. Method: To study the crystallographic aspects of iron plaque formation on rice roots, the concentrations of Fe2+ and PO4 3- were controlled in this study. The effects of these treatments were assessed through comprehensive analyzes encompassing root growth status, root surface iron concentration, root vitality, enzyme activities, and microstructural characteristics using advanced techniques such as root analysis, scanning electron microscopy (SEM), and ultrathin section transmission electron microscopy (TEM). Results: The results demonstrated that an increase in the Fe2+ concentration or a decrease in the PO4 3- concentration in the nutrient solution led to improvements in various root growth indicators. There was an elevation in the DCB (dithionite-citrate-bicarbonate) iron content within the roots, enhanced root vitality, and a significant increase in the activities of the superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) enzymes. Moreover, as the Fe2+ concentration increased, amorphous iron oxide minerals on the root surface were gradually transformed into ferrihydrite particles with sizes of approximately 200 nm and goethite particles with sizes of approximately 5 µm. This study showed that an increase in the Fe2+ concentration and a decrease in the PO4 3- concentration led to the formation of substantial iron plaque on the root surfaces. It is noteworthy that there was a distinct gap ranging from 0.5 to 3 µm between the iron plaque formed through PO4 3- treatment and the cellular layer of the root surface. Discussion: This study elucidated the impacts of Fe2+ and PO4 3- treatments on the formation, structure, and morphology of the iron plaque while discerning variations in the spatial proximity between the iron plaque and root surface under different treatment conditions.
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Self-assembled functional nanomaterials with electromagnetic hot spots are crucial and highly desirable in surface-enhanced Raman scattering (SERS). Due to its versatile biological scaffold, the M13 phage has been employed to produce novel nano-building blocks and devices. In this study, we propose a novel M13 phage-based SERS nanocarrier, that utilizes the pVIII capsid in M13 to conjugate Au@Ag core-shell nanorod (Au@AgNR) with linker carboxy-PEG-thiol (M13-Au@AgNR) and the pIII capsid to specifically target Escherichia coli (E. coli). The M13-Au@AgNR@DTTC (3,3'- diethylthiocarbocyanine iodide) SERS probe was used to detect E. coli in a concentration range of 6 to 6 × 105 cfu/mL, achieving a limit of detection (LOD) of 0.5 cfu/mL. The proposed SERS platform was also tested in real samples, showing good recoveries (92%-114.3%) and a relative standard deviation (RSD) of 1.2%-4.7%. Furthermore, the system demonstrated high antibacterial efficiency against E. coli, approximately 90%, as measured by the standard plate-count method. The investigation provides an effective strategy for in vitro bacteria detection and inactivation.
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This study focuses on understanding the growth and control of the gold nanoplates by seed-mediated growth approach. These monodispersive size-controlled gold nanoplates have the average thickness of 8-10 nm and average size tunable from 70 to 150 nm, exhibiting strong surface plasmon absorption in the near infrared (NIR) region. For the gold nanoplates formation, the seeds serve as nucleation sites, ascorbic acid (AA) serves as a new reductant to reduce hydrogen tetrachloroaurate (HAuCl4), surface activity system cetyltrimethylammonium bromide (CTAB) and potassium iodide (KI) are critical factors. X-ray diffraction (XRD) and selected-area electron diffraction (SAED) analyses reveal that gold nanoplates with the (111) lattice plane as the basal plane are single crystals. CTAB are absorbed on the surface of the (111) lattice plane of the single crystals, accounting for self-assembled monolayer and head-to-head arrays. The two arrays have been shown to serve as effective surface-enhanced Raman scattering substrates using Niel blue A (NBA) sulfate as Raman report molecule.
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Cristalização/métodos , Ouro/química , Nanoestruturas/química , Nanoestruturas/ultraestrutura , Análise Espectral Raman/métodos , Substâncias Macromoleculares/química , Teste de Materiais , Conformação Molecular , Tamanho da Partícula , Eletricidade Estática , Estresse Mecânico , Propriedades de SuperfícieRESUMO
The ability to monitor the differentiation of living stem cells is essential for understanding stem cell biology and the practical application of stem cell therapies. However, conventional methods of analyzing biomarkers related to differentiation still require a large number of cells or cell lysates. This requirement leads to the unavoidable loss of cell sources and hinders the real-time monitoring of cellular processes. In this study, we report an ultrasensitive surface-enhanced Raman scattering (SERS) method for the long-term detection and imaging of miR-144-3p in osteogenic differentiation of BMSCs, by using target miRNA-induced gold nanocage (GNC)-hairpin DNA1 (hpDNA1)-hpDNA2-GNC assembly in living cells. The finite-difference time domain method demonstrated that the electromagnetic intensities of the dimer and polymer of the GNCs were significantly enhanced compared to that of GNCs only, which theoretically confirmed the rational design of the SERS strategy. The hpDNA-conjugated GNC probes were prepared and used to recognize the target and distinguish from other miRNAs. This method enabled excellent sensitivity and high selectivity toward miR-144-3p with a limit of detection of 13.6 aM and a broad range from 100 aM to 100 pM in cell lysates. Then, we used transmission electron microscopy images, fluorescence microscopy images, and dark-field microscopy images to study the internalization of the probes in BMSCs. A Cell Counting Kit-8 experiment indicated that the probes were not cytotoxic in a certain concentration range. BMSCs were treated with an osteogenic inductor so that they would subsequently differentiate into osteocytes. Upon cellular uptake of these nanoprobes, we observed intense and time-dependent SERS responses from the important osteogenic biomarker miR-144-3p, only in BMSCs undergoing osteogenic differentiation and living undifferentiated BMSCs but not in osteoblasts. Finally, the accuracy of SERS has been proved by a quantitative real-time polymerase chain reaction experiment. The above results demonstrated that our nanoprobes are capable of long-term tracking of the dynamic expression of miR-144-3p (21 days) in the differentiating BMSCs. SERS has broad application prospects in the long-term detection of stem cell differentiation, and identification and isolation of specific cell types as well as in biomedical diagnosis.
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Células da Medula Óssea , Diferenciação Celular , Ouro , Nanopartículas Metálicas/química , MicroRNAs/metabolismo , Imagem Molecular , Sondas Moleculares , Análise Espectral Raman , Células-Tronco , Animais , Células da Medula Óssea/citologia , Células da Medula Óssea/metabolismo , Ouro/química , Ouro/farmacologia , Masculino , Sondas Moleculares/química , Sondas Moleculares/farmacologia , Ratos , Ratos Sprague-Dawley , Células-Tronco/citologia , Células-Tronco/metabolismoRESUMO
Tumor resistance and drug-induced nephrotoxicity pose great challenges to the clinical treatment of tumors, and they also limit the clinical application of oncology drugs. Finding an effective adjuvant, which can sensitize tumor treatment, is an effective method for tumor treatment. Here, we developed a ratiometric fluorescent probe, TP-Tfs, for superoxide anion (O2Ë-) detection in living cells and in vivo during the process of tumor treatment for the first time. TP-Tfs with simple synthesis steps and high yields can detect O2Ë- sensitively and selectively, and the detection limit was determined to be 37 nM. Using TP-Tfs, we found that cis-diaminodichloroplatinum(ii) (DDP) was effective in treating tumors by inducing O2Ë- burst. Curcumin (cum) can sensitize tumor treatment effectively by inducing more severe O2Ë- burst. These results indicated that the probe TP-Tfs was a promising candidate for drug screening and tumor treatment evaluation.
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Antineoplásicos/farmacologia , Cisplatino/farmacologia , Corantes Fluorescentes/química , Neoplasias Pulmonares/tratamento farmacológico , Neuroblastoma/tratamento farmacológico , Superóxidos/análise , Ânions/análise , Antineoplásicos/química , Linhagem Celular Tumoral , Cisplatino/química , Corantes Fluorescentes/síntese química , Humanos , Neoplasias Pulmonares/diagnóstico por imagem , Teste de Materiais , Estrutura Molecular , Neuroblastoma/diagnóstico por imagem , Imagem Óptica , Tamanho da Partícula , Propriedades de SuperfícieRESUMO
The accurate analysis of tumor related biomarkers is extremely critical in the diagnosis of the early stage cervical cancer. Herein, we designed a novel and inexpensive surface-enhanced Raman scattering-based lateral flow assay (SERS-based LFA) strip with a single test line, which was applied for the rapid and sensitive quantitative simultaneous analysis of SCCA and CA125 in serum samples from patients with cervical cancer. In the presence of target antigens, the monoclonal antibody-coupled and Raman reporter-labeled nano-Ag polydopamine nanospheres (PDA@Ag-NPs) aggregated on the test line modified by the polyclonal antibody to form a double-antibody sandwich structure. The finite difference time domain simulation demonstrated that large number of "hot spots" was generated among the nanogaps of aggregated PDA@AgNPs, which resulted in a huge enhancement of the signal of the Raman reporters. Accordingly, the limit of detection was determined to be 7.156 pg mL-1 for SCCA and 7.182 pg mL-1 for CA125 in phosphate buffer and 8.093 pg mL-1 for SCCA and 7.370 pg mL-1 for CA125 in human serum, revealing high sensitivity of this SERS-based LFA strip. Significantly, the detection of SCCA and CA125 using the SERS-based LFA was observed to have high specificity and reproducibility, and the whole detection was completed within 20 min. Furthermore, the SERS-based LFA and enzyme-linked immunosorbent assay were also employed in serum samples obtained from patients with cervical cancer, cervical intraepithelial neoplasia and healthy subjects, and perfect agreement existed between both the methods. Thus, clinically, the developed SERS-based LFA strip has strong potential for the simultaneous detection of multiple cancer biomarkers in serum.
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This minireview reports the recent advances in surface-enhanced Raman scattering (SERS)-based assay devices for the diagnosis of infectious diseases. SERS-based detection methods have shown promise in overcoming the low sensitivity and multiplex detection problems inherent to fluorescence detection. Therefore, it is interesting to investigate the current status, challenges, and applications associated with SERS-based microdevices for the point-of-care (POC) diagnosis of infectious diseases. The majority of this review highlights three different types of microdevices, namely microfluidic channels, lateral flow assay strips, and three-dimensional nanostructured substrates. Furthermore, the integration of portable Raman spectrophotometry with microdevices provides an ideal platform for the diagnosis of various infectious diseases in the field. Integrated SERS-based assay systems also enable measurements in minimal sample volumes and at low analyte concentrations of viral or bacterial samples. A significant number of studies using the SERS-based assay system have been performed recently to realize POC diagnostics, especially under resource-limited conditions. This portable SERS sensor is expected to be a next-generation POC assay system that could overcome the limitations of current fluorescence-based assay systems. This minireview summarizes recent advances in the development of SERS-based microdevices for the diagnosis of infectious diseases. Lastly, challenges to overcome and future perspectives are discussed.
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Nanoestruturas , Sistemas Automatizados de Assistência Junto ao Leito , Vírus , Bactérias , Análise Espectral RamanRESUMO
There is a challenge to obtain an ultrasensitive and rapid approach to detect bacteria and identify resistance. As a powerful bioanalytical tool, surface-enhanced Raman scattering (SERS) in bacterial detection have attracted increasing attentions. Herein, we developed a SERS-active Au@Ag core-shell nanorod (Au@AgNR) tag platform for ultrasensitive bacteria detection and antibiotic-susceptibility testing (AST). The platform established that surface enhanced Raman scattered Rhodamine 6G (R6G) absorption at 1517 cm-1 had a good linearity (RI = 3865 + 193logC; R2 = 0.97) with logarithm of E. coli concentration over a range of 107-102 CFU (colony forming unit)/mL with limit of detection as low 102 CFU/mL. When E. coli was exposed to ampicillin at minimum inhibitory concentration (MIC, 4 µg/mL), Raman spectroscopy showed the obvious variation between ampicillin-susceptible E. coli (Amp--E. coli) and the ampicillin-resistant E. coli (Amp+-E. coli). Combined with principal component analysis (PCA) statistical analysis, the Raman intensity variation mentioned above allows to obtain rapid antibiotic resistance testing (<3.5 h). In addition, E.coli spiked into blood from C57BL/6 mice can be identified clearly, indicating the potential for point-of-care diagnostics.
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Nanotubos , Prata , Animais , Antibacterianos/farmacologia , Bactérias , Escherichia coli , Ouro , Camundongos , Camundongos Endogâmicos C57BL , Análise Espectral RamanRESUMO
Fatty acids (FAs) are potential therapeutic agents for cutaneous wound healing; however, the mechanisms underlying this effect have not been clearly defined. In this study, we extracted and characterized FAs from dried Lucilia sericata larvae and investigated the molecular basis by which FAs promote cutaneous wound healing. We first confirmed that FA sodium salts (FASSs) stimulated proliferation, migration, and tube formation of cultured human umbilical vein endothelial cells (HUVECs) in a dose-dependent manner. We then showed that FASSs promoted endothelial-to-mesenchymal transition (EndMT), which plays an important role in stabilizing the neovasculature during angiogenesis. Mechanistically, FASSs up-regulated the expression of angiogenesis-related growth factors, platelet-derived growth factor (PDGF), transforming growth factor-ß1 (TGF-ß1), and vascular endothelial growth factor A (VEGFA), and activated angiogenesis-related signaling pathways, AKT, ERK, and TGF-ß/Smad3. In a rat acute cutaneous-wound model, FAs promoted wound healing. Following treatment, we further found that expression of anti-apoptosis-related factors (c-Myc and Bcl-2) was up-regulated and expression of apoptosis-related factors (p53 and Bad) was down-regulated. Our findings suggest that FAs can promote cutaneous wound healing by inducing angiogenesis, partly by activating AKT, ERK, and TGF-ß/Smad3 signaling.
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In the ascending auditory pathway, the central nucleus of the inferior colliculus (IC) receives and integrates excitatory and inhibitory inputs from many bilateral lower auditory nuclei, intrinsic projections within the IC, contralateral IC through the commissure of the IC and from the auditory cortex. All these presynaptic excitatory and inhibitory inputs dynamically shape and modulate the auditory response properties of individual IC neurons. For this reason, acoustic response properties vary among individual IC neurons due to different activity pattern of presynaptic inputs. The present study examines modulation of auditory response properties of IC neurons by combining sound stimulation with focal electrical stimulation of the contralateral dorsal nucleus of the lateral lemniscus (referred to as ESDNLL) in the albino mouse. Brief ESDNLL produces variation (increase or decrease) in the number of impulses, response latency and discharge duration of modulated IC neurons. Additionally, 30-minute short-term ESDNLL alone produces variation in the best frequency (BF) and minimum threshold (MT) of modulated IC neurons. These varied response parameters recover in different manner and time course among individual modulated IC neurons. Possible pathways and neural mechanisms underlying these findings are discussed.
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Vias Auditivas/fisiologia , Percepção Auditiva/fisiologia , Colículos Inferiores/fisiologia , Neurônios/fisiologia , Estimulação Acústica , Potenciais de Ação , Adaptação Fisiológica , Animais , Estimulação Elétrica , Feminino , Masculino , CamundongosRESUMO
Conventional research on surface-enhanced Raman scattering (SERS)-based pH sensors often depends on nanoparticle aggregation, whereas the variability in nanoparticle aggregation gives rise to poor repeatability in the SERS signal. Herein, we fabricated a gold nanorod array platform via an efficient evaporative self-assembly method. The platform exhibits great SERS sensitivity with an enhancement factor of 5.6 × 107 and maintains excellent recyclability and reproducibility with relative standard deviation (RSD) values of less than 8%. On the basis of the platform, we developed a highly sensitive bovine serum albumin (BSA)-coated 4-mercaptopyridine (4-MPy)-linked (BMP) SERS-based pH sensor to report pH ranging from pH 3.0 to pH 8.0. The intensity ratio variation of 1004 and 1096 cm-1 in 4-MPy showed excellent pH sensitivity, which decreased as the surrounding pH increased. Furthermore, this BMP SERS-based pH sensor was employed to measure the pH value in C57BL/6 mouse blood. We have demonstrated that the pH sensor has great advantages such as good stability, reliability, and accuracy, which could be extended for the design of point-of-care devices.
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Nanotubos , Animais , Ouro , Concentração de Íons de Hidrogênio , Nanopartículas Metálicas , Camundongos , Camundongos Endogâmicos C57BL , Reprodutibilidade dos Testes , Análise Espectral RamanRESUMO
We investigated the effect of miR-182-5p on the viability, proliferation, invasion, and migration ability of human gastric cells by regulating the expression of RAB27A. Real-time PCR assay was used to detect the expression of miR-182-5 and RAB27A in human gastric carcinoma tissues, para-carcinoma tissues, and different cell lines. Western blotting was also used to determine the RAB27A expression in both tissues and cell lines. We chose the HGC-27 cell line as experiment subject as it demonstrated the highest miR-182-5p level. HGC-27 cells were transfected with different vectors and the cell viability, mitosis, invasion, and migration ability were measured through MTT assay, flow cytometry (FCM) analysis, Transwell assay, and wound healing assay. In comparison with the normal tissues, miR-182-5p is expressed at a higher level in gastric cancer (GC) tissues, while RAB27A is expressed at a lower level in cancerous tissues. The down-regulation of miR-182-5p and up-regulation of RAB27A can significantly decrease the viability, migration, invasion, and mitosis of HGC-27 cells. The target relationship between miR-182-5p and RAb27A was confirmed through a dual-luciferase reporter gene assay and Western blot assay. miR-182-5p enhances the viability, mitosis, migration, and invasion of human GC cells by down-regulating RAB27A.
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Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , Invasividade Neoplásica/genética , Neoplasias Gástricas/genética , Proteínas rab27 de Ligação ao GTP/genética , Linhagem Celular Tumoral , Movimento Celular , Sobrevivência Celular , Regulação para Baixo , Humanos , Mitose , Invasividade Neoplásica/patologia , Neoplasias Gástricas/patologiaRESUMO
OBJECTIVE: To investigate the protective effect of glutamine in rats with pseudomonas pneumonia receiving total parenteral nutritional (TPN) therapy. METHODS: Forty Sprague-Dawley rats were randomly divided into 4 groups (Group A, B, C and D). Rats in Group A received, by intra-tracheal route, infusion of 0.3 ml normal saline. Rats in Group B received, also by intra-tracheal route, infusion of 0.3 ml pseudomonas aeruginosa soliquoid so that the bacteria entered the lungs directly. A standard TPN solution not containing glutamine was administered intravenously to rats in Group C (160 ml/kg) for 5 days, and a TPN solution containing glutamine was administered intravenously to rats in Group D (160 ml/kg) for 5 days. On day 6, 0.3 ml pseudomonas aeruginosa soliquoid was administered by intra-tracheal route to rats in Group C and D. The weight of rats, their activity and mortality were recorded. 48 h after intratracheal administration, blood and bronchoalveolar lavage fluid (BALF) samples were collected for measurement of white blood cell count, TNFalpha, IL-1, IL-10 and total protein of BALF. Segments of the lung, the liver and the ileum tissues were collected for HE pathological slice. RESULTS: (1) Initially there was no difference in weight between the four groups, but on day 8 the weights of rats in Group B, C, and D were significantly decreased compared with that in Group A (P < 0.05). The mortality of rats in Group C was significantly higher than that in Group B and D (P < 0.05). (2) There were significant differences between Group C and D in white blood cell count, blood TNFalpha and IL-10 level, BALF, and total protein of BALF (P < 0.05). (3) Pathological changes of the lung, the liver and ileum in Group C were more severe than that in Group B and D. Group A was basically normal. CONCLUSION: Glutamine is able to protect gastrointestinal tract function in rats receiving TPN, to improve pulmonary anti-infection ability, and to alleviate injuries of important organs caused by severe infection.
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Glutamina/farmacologia , Pneumonia Bacteriana/prevenção & controle , Animais , Trato Gastrointestinal/efeitos dos fármacos , Íleo/patologia , Fígado/patologia , Pulmão/patologia , Nutrição Parenteral Total , Pneumonia Bacteriana/patologia , Pseudomonas aeruginosa , Distribuição Aleatória , Ratos , Ratos Sprague-DawleyRESUMO
A surfactant-free and template-free method for the high-yield synthesis of biomolecule (serotonin)-based formaldehyde resin (BFR) microspheres is proposed for the first time. The colloidal microspheres loaded with Au nanoparticles (AuNPs) prepared by a convenient in-situ synthesis of AuNPs on BFR (AuNPs/BFR) microsphere surface show good stability. AuNPs/BFR microspheres not only favor the immobilization of antibody but also facilitate the electron transfer. It is found that the resultant AuNPs/BFR microspheres can be designed to act as a sensitive label-free electrochemical immunosensor for carcinoembryonic antigen (CEA) determination. The immunosensor is prepared by immobilizing capture anti-CEA on AuNPs/BFR microspheres assembled on thionine (TH) modified glassy carbon electrode (GCE). TH acts as the redox probe. Under the optimized conditions, the linear range of the proposed immunosensor is estimated to be from 25 pg/mL to 2000 pg/mL (R=0.998) and the detection limit is estimated to be 3.5 pg/mL at a signal-to-noise ratio of 3. The prepared immunosensor for detection of CEA shows high sensitivity, reproducibility and stability. Our study demonstrates that the immunosensor can be used for the CEA detection in humans serum.
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Biomarcadores Tumorais/isolamento & purificação , Técnicas Biossensoriais/métodos , Antígeno Carcinoembrionário/isolamento & purificação , Biomarcadores Tumorais/sangue , Antígeno Carcinoembrionário/sangue , Quitosana/química , Formaldeído/química , Ouro/química , Humanos , Imunoensaio/métodos , Limite de Detecção , Nanopartículas Metálicas/química , MicroesferasRESUMO
The silver-modified gold nanoplate arrays as bimetallic surface-enhanced Raman scattering (SERS) substrates were optimized for the surface-enhanced Raman detection of streptavidin/biotin monolayer assemblies. The bimetallic gold-silver nanoplate arrays were fabricated by coating silver nanoparticles uniformly on the gold nanoplate arrays. Depending on silver nanoparticle coating, the localized surface plasmon resonance (LSPR) peak of the bimetallic gold-silver nanoplate arrays blue-shifted and broadened significantly. The common probe molecule, Niel Blue A sulfate (NBA) was used for testing the SERS activity of the bimetallic gold-silver nanoplate arrays. The SERS intensity increased with the silver nanoparticle coating, due to a large number of hot spots and nanoparticle interfaces. The platforms were tested against a monolayer of streptavidin functionalized over the bimetallic gold-silver nanoplate arrays showing that good quality spectra could be acquired with a short acquisition time. The supramolecular interaction between streptavidin (strep) and biotin showed subsequent modification of Raman spectra that implied a change of the secondary structure of the host biomolecule. And the detection concentration for biotin by this method was as low as 1.0 nM. The enhanced SERS performance of such bimetallic gold-silver nanoplate arrays could spur further interest in the integration of highly sensitive biosensors for rapid, nondestructive, and quantitative bioanalysis, particularly in microfluidics.