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BACKGROUND AND AIMS: Atherosclerosis preferentially occurs at regions in arterial branching, curvature, and stenosis, which may be explained by the geometric predilection of low-density lipoprotein (LDL) concentration polarization that has been investigated in major arteries in previous studies. Whether this also happens in arterioles remains unknown. METHODS: Herein, a radially non-uniform distribution of LDL particles and a heterogeneous endothelial glycocalyx layer in the mouse ear arterioles, as shown by fluorescein isothiocyanate labeled wheat germ agglutinin (WGA-FITC), were successfully observed by a non-invasive two-photon laser-scanning microscopy (TPLSM) technique. The stagnant film theory was applied as the fitting function to evaluate LDL concentration polarization in arterioles. RESULTS: The concentration polarization rate (CPR, the ratio of the number of polarized cases to that of total cases) in the inner walls of curved and branched arterioles was 22% and 31% higher than the outer counterparts, respectively. Results from the binary logistic regression and multiple linear regression analysis showed that endothelial glycocalyx thickness increases CPR and the thickness of the concentration polarization layer (CPL). Flow field computation indicates no obvious disturbances or vortex in modeled arterioles with different geometries and the mean wall shear stress is about 7.7-9.0 Pa. CONCLUSIONS: These findings suggest a geometric predilection of LDL concentration polarization in arterioles for the first time, and the existence of an endothelial glycocalyx, acting together with a relatively high wall shear stress in arterioles, may explain to some extent why atherosclerosis rarely occurs in these regions.
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Aterosclerosis , Lipoproteínas LDL , Animales , Ratones , Arteriolas , Glicocálix , ArteriasRESUMEN
Most small diameter vascular grafts (inner diameter<6 mm) evaluation studies are performed in healthy animals that cannot represent the clinical situation. Herein, an hypercholesterolemia (HC) rat model with thickened intima and elevated expression of pro-inflammatory intercellular adhesion molecular-1 (ICAM-1) in the carotid branch is established. Electrospun polycaprolactone (PCL) vascular grafts (length: 1 cm; inner diameter: 2 mm) are implanted into the HC rat abdominal aortas in an end to end fashion and followed up to 43 days, showing a relative lower patency accompanied by significant neointima hyperplasia, abundant collagen deposition, and slower endothelialization than those implanted into healthy ones. Moreover, the proliferation, migration, and adhesion behavior of endothelial cells (ECs) isolated from the HC aortas are impaired as evaluated under both static and pulsatile flow conditions. DNA microarray studies of the HC aortic endothelium suggest genes involved in EC proliferation (Egr2), apoptosis (Zbtb16 and Mt1), and metabolism (Slc7a11 and Hamp) are down regulated. These results suggest the impaired proliferative, migratory, and adhesive abilities of ECs are associated with the bad performances of grafts in HC rat. Future pre-clinical evaluation of small diameter vascular grafts may concern more disease animal models with clinical complications. STATEMENT OF SIGNIFICANCE: During the development of small diameter vascular grafts (D<6 mm), young and healthy animal models from pigs, sheep, dogs, to rabbits and rats are preferred. However, it cannot represent the clinic situation, where most cardiovascular grafting procedures are performed in the elderly and age is the primary risk factor for disease development or death. Herein, the performance of electrospun polycaprolactone (PCL) vascular grafts implanted into hypercholesterolemia (HC) or healthy rats were evaluated. Results suggest the proliferative, migratory, and adhesive abilities of endothelial cells (ECs) are already impaired in HC rats, which contributes to the observed slower endothelialization of implanted PCL grafts. Future pre-clinical evaluation of small diameter vascular grafts may concern more disease animal models with clinical complications.
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Células Endoteliales , Hipercolesterolemia , Adhesivos , Animales , Prótesis Vascular , Proliferación Celular , Perros , Poliésteres , Conejos , Ratas , Ovinos , PorcinosRESUMEN
In this study, green low-toxicity Mn-doped Zns (Mn-Zns) room-temperature phosphorescent (RTP) quantum dots (QDs) (PQDs) nanocomposites (DNA-PQDs) were prepared under the optimal conditions by using single-stranded DNA (PS-C-ssDNA) rich of cytosine C and Thioguanine G (PS) as the template. DNA-PQDs interact with Ag+ to form AgN bonds and further produce C-Ag+-C conjugates. As a result, DNA-PQDs cluster together and induce the phosphorescent exciton energy transfer (PEET), resulting in quenching of room-temperature phosphorescent of DNA-PQDs. Nevertheless, Ag+ tends to form AgS bonds with biological mercaptan when it is added in, so that Ag+ falls from C-Ag+-C. DNA-PQDs changed from aggregation to looseness and RTP is recovered accordingly. On this basis, RTP detection of biological mercaptan is realized. Since this sensor system has RTP properties based on DNA-PQDs, it is very applicable to detection of mercaptan compounds in biological fluids.
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ADN de Cadena Simple/química , Sustancias Luminiscentes/química , Manganeso/química , Nanocompuestos/química , Puntos Cuánticos/química , Compuestos de Sulfhidrilo/análisis , Técnicas Biosensibles/métodos , Transferencia de Energía , Humanos , Límite de Detección , Modelos Moleculares , Compuestos de Sulfhidrilo/sangre , Zinc/químicaRESUMEN
Alizarin Red S (ARS) has been extensively used in the dyeing industry. In order to effectively remove the ARS form dyeing wastewater, polyethyleneimine (PEI)-functionalized magnetic carbon nanotubes (PEI@MCNTs) adsorbent was successfully prepared and its adsorption performances were also investigated in detail. The PEI@MCNTs could efficiently remove the ARS from acidic aqueous solution (pHâ¯≤â¯6.0) within 40â¯min under room temperature. Benefiting from a large number of adsorption sites and multiple interactions, PEI@MCNTs possessed high selectivity towards ARS with spontaneous adsorption process. The maximum adsorption capacity of PEI@MCNTs for ARS was 196.08â¯mgâ¯g-1 obtained from Langmuir isotherm, higher than that of available conventional adsorbents. Moreover, the PEI@MCNTs could be easily collected by an external magnet, and then effectively regenerated through 10â¯mM NaOH solution. The prepared PEI@MCNTs could be considered as the promising adsorbent for the removal of anthraquinone dyes in large-scale wastewater treatment.
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Nanotubos de Carbono , Contaminantes Químicos del Agua , Adsorción , Antraquinonas , Cinética , PolietileneiminaRESUMEN
A one-pot hydrothermal method was developed for the synthesis of graphene oxide quantum dots (GOQDs). It is making use of toner waste as the precursor and H2O2 as the oxidant. Synthesis takes 4 h and does not require strong acids or complex purification steps and does not produce environmentally harmful metal ions. The GOQDs display blue fluorescence with excitation/emission maxima at 340/445 nm. The feasibility of detecting specific DNA sequence was promoted using polyethyleneimine to modify the GOQDs surface. A method was developed to recognized a specific DNA sequence. This is based on electrostatic aggregation of GOQDs and ssDNA labeled with Dabcyl at the 3' end, which promotes fluorescence quenching of GOQDs. The possible fluorescence quenching mechanism (which is mainly dynamic) was investigated using the Stern-Volmer equation. When a target sequence was added, which is complementary to the ssDNA, the dabcyl-labeled ssDNA is released due to strict complementary base pairing. This promotes fluorescence recovery of GOQDs. The assay has a 0.17 nM detection limit and a linear range of 0.5-30 nM. The method was used to quantify specific DNA sequences from extracts of genetically modified plant tissues. Graphical abstract Graphene oxide quantum dots (GOQDs) were synthesized by one-pot hydrothermal method using waste toner, and the surface was modified by polyethyleneimine (PEI). Through the interaction of PEI-GOQDs with Dabcyl-DNA single strands to dynamically quench the fluorescence of GOQDs. Based on DNA hybridization technology, we established specific DNA sequence detection nanoprobe.
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ADN de Cadena Simple/análisis , Colorantes Fluorescentes/química , Grafito/química , Puntos Cuánticos/química , Procesos de Copia , Sondas de ADN/química , Sondas de ADN/genética , ADN de Cadena Simple/química , ADN de Cadena Simple/genética , Colorantes Fluorescentes/síntesis química , Peróxido de Hidrógeno/química , Límite de Detección , Hibridación de Ácido Nucleico , Plantas/química , Polietileneimina/química , Espectrometría de Fluorescencia/métodos , ResiduosRESUMEN
Mesoporous molecularly imprinted polymers (MIPs) containing mangnanese-doped ZnS quantum dots (Mn-ZnS QDs) were prepared for specific recognition and detection of 4-nitrophenol (4-NP). The Mn-ZnS QDs display orange room-temperature phosphorescence with excitation/emission peaks at 295/590 nm and a decay time of 2.0 ms. In the presence of 4-NP, the orange phosphorescence is strongly reduced. Phosphorescence drops linearly in the 0.1-100 µM 4-NP concentration range, and the detection limit is 60 nM. The detection limit is far lower than the maximally allowed 4-NP concentrations in surface water and drinking water as specified by the U.S. Environmental Protection Agency. The intraday (n = 5) and interday (n = 6) spiked recovery rates were 96.0-104.5% and 97.9-107.9%, respectively, with relative standard deviations of 0.7-4.8% and 1.8-7.5% respectively. These MIPs integrated the characteristic features of phosphorimetry and molecular imprinting. Potential interference by competitive substances, background fluorescence or scattered light are widely reduced. Graphical abstract Schematic presentation of the synthesis of phosphorescent molecularly-imprinted polymers. A novel probe with manganese-doped ZnS quantum dots (Mn-ZnS QDs) and 3-aminopropyl-triethoxysilane (APTES) as functional monomers and tetraethoxysilane (TEOS) as crosslinking agent was prepared for selective phosphorescence detection of 4-nitrophenol (4-NP).
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The direction synthesis of biofunctional nanomaterials with DNA as the template is of high application value. By using phosphorothioate-thymine single-stranded DNA (PS-T-ssDNA) as the template and through synthetic conditions optimization, novel low-toxicity and environment-friendly ssDNA-functionalized room-temperature phosphorescent quantum dots (PS-T-ssDNA RTP QDs) were prepared at low temperature (37 °C). Then, the quantitative RTP-based mercury(II) (Hg2+) detection was achieved by utilizing the specific identifying ability of T-base-pair Hg2+ (T-Hg2+-T) and its photoinduced electron transfer. This RTP sensor in Hg2+ detection had a linear range of 0.02 to 0.8 µM and a detection limit of 4.8 nM. The dependence on RTP of QDs effectively avoids interference from background fluorescence and scattering light in the environment or biological samples. This sensor also possessed an RTP stability and a long service life and did not require sample pretreatment. Thus this sensor is suitable for environmental and quantitative Hg2+ detection in biological samples.
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Técnicas Biosensibles/métodos , ADN de Cadena Simple/química , Ambiente , Sustancias Luminiscentes/química , Mercurio/análisis , Puntos Cuánticos/química , Temperatura , Mercurio/orina , Modelos Moleculares , Conformación MolecularRESUMEN
The relationship between the thickness of surface molecularly imprinted polymers (MIPs) and specific recognition performance of transferrin (Trf) as well as the quantitative relation between the grafting amount of Mn-ZnS room-temperature phosphorescence (RTP) quantum dots (QDs) (short for PQDs) and RTP signals for recognition of Trf was analyzed in this study. Based on analysis results, RTP protein mesoporous imprinting microspheres (SiO2-PQDs-MIPs) with high specificity and strong interference resistance were developed using a mesoporous SiO2 nanomaterial that can create more three-dimensional precise recognition sites as the matrix and using PQDs with strong resistance to background fluorescence interference as the luminescent materials. A discriminatory analysis of Trf was realized by the phosphorescence quenching principle based on light quenching caused by the photoinduced electron transfer. The concentration range, limit of detection, relative standard deviation, and imprinting factor of Trf detection under pH 7.4 are 0.05-1.0 µM, 0.014 µM, 3.23%, and 3.09, respectively. Although the sensing signals of SiO2-PQDs-MIPs for proteins are based on the phosphorescence of PQDs, they are particularly suitable for specific recognition and accurate quantitative detection of proteins in biological fluids. Research conclusions are expected to realize high-efficiency recognition of target proteins in actual biological samples.
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Líquidos Corporales/metabolismo , Microesferas , Puntos Cuánticos/química , Dióxido de Silicio/química , Transferrina/análisis , Animales , Humanos , Límite de Detección , Transferrina/metabolismoRESUMEN
Nonalcoholic fatty liver disease (NAFLD) is a major risk factor of many end-stage liver diseases. Alterations in microRNA expression have been reported in patients with NAFLD. However, the transcriptional mechanism(s) of dysregulated microRNAs under the state of NAFLD is poorly described, and microRNAs that regulate the pathogenesis of NAFLD synergistically with their regulators remain unknown. Here we report that microRNA-378 expression is significantly increased in fatty livers of mice and patients with NAFLD. Although microRNA-378 locates within the intron of Ppargc1ß (peroxisome proliferator-activated receptor γ coactivator 1-beta), there was a significant uncoupling of Ppargc1ß mRNA and microRNA-378 levels in both sources of fatty livers. Further studies identified a full-length primary transcript of microRNA-378. LXRα (liver X receptor alpha) functioned as a transcription activator of microRNA-378 and a repressor of Ppargc1ß transcription. It is known that miR-378 is an inhibitor of fatty acid oxidation (FAO) and the function of Ppargc1ß is opposite to that of miR-378. GW3965 treatment (LXRα agonist) of murine hepatocytes and mice increased microRNA-378 and reduced Ppargc1ß, which subsequently impaired FAO and aggravated hepatosteatosis. In contrast, additional treatment of miR-378 inhibitor or Ppargc1ß, which knocked down increased miR-378 or recovered expression of Ppargc1ß, offset the effects of GW3965. Liver-specific ablation of Lxrα led to decreased miR-378 and increased Ppargc1ß, which subsequently improved FAO and reduced hepatosteatosis. Conclusion: Our findings indicated that miR-378 possesses its own transcription machinery, which challenges the well-established dogma that miR-378 transcription is controlled by the promoter of Ppargc1ß. LXRα selectively activates transcription of miR-378 and inhibits expression of Ppargc1ß, which synergistically impairs FAO. In addition to lipogenesis, impaired FAO by miR-378 in part contributes to LXRα-induced hepatosteatosis.
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Hígado Graso/etiología , Receptores X del Hígado/metabolismo , MicroARNs/biosíntesis , Proteínas Nucleares/metabolismo , Factores de Transcripción/metabolismo , Animales , Benzoatos , Bencilaminas , ARN Helicasas DEAD-box/metabolismo , Hígado Graso/metabolismo , Regulación de la Expresión Génica , Células Hep G2 , Humanos , Metabolismo de los Lípidos , Hígado/metabolismo , Receptores X del Hígado/agonistas , Masculino , Ratones Endogámicos C57BL , Proteínas de Unión al ARN/metabolismo , Ribonucleasa III/metabolismoRESUMEN
A simple and sensitive Mn-ZnS quantum dot room-temperature phosphorescent immunosensor for detecting microcystin-LR was developed. This sensor adopted antigens and antibodies as recognition units and used Mn-ZnS RTP QDs as sensing materials to specifically bind with MC-LR. The structurally specific binding between the microcystin-LR antibody and MC-LR led to the aggregation of antibody-crosslinked QDs, and then the electrons of QDs would be transferred to the complex, leading to the phosphorescence quenching of QDs. The microcystin-LR antigen-antibody specific binding site was first analyzed. This phosphorescent immunosensor rapidly and sensitively detected microcystin-LR, with linear ranges of 0.2-1.5 µg L-1 and 1.5-20 µg L-1 and a detection limit of up to 0.024 µg L-1. Meanwhile, coexisting pollutants of microcystin-LR in water did not significantly interfere with microcystin-LR detection. The new sensor was applied to detect real water samples and showed high sensitivity and selectivity.
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Bio-functionalization of quantum dots (QDs) is of important value in practical applications. With single-stranded DNA (ssDNA) rich in thymine T and thioguanine G taken as the template, a new-type nanocomposite material (ssDNA-PQDs) synthesized from low-toxicity T-ssDNA functionalized Mn-ZnS and room-temperature phosphorescent (RTP) QDs (PQDs) was prepared in this paper by optimizing synthesis conditions, and these ssDNA-PQDs could emit orange RTP signals at 590 nm. As these ssDNA-PQDs are rich in T sequences and T sequences can bond with melamine through the hydrogen-bond interaction, ssDNA-PQDs experience aggregation, thus causing phosphorescent exciton energy transfer (PEET) between ssDNA-PQDs of different particle sizes and their RTP quenching. Based on this principle, an RTP detection method for melamine was established. The linear range and detection limit of the detection method are 0.005-6 mM and 0.0016 mM respectively. As this method is based on the RTP nature of ssDNA-PQDs, it can avoid disturbance from background fluorescence and scattered light of the biological fluid, and it is very suitable for melamine detection in the biological fluid milk.
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Given the outstanding room-temperature phosphorescence (RTP) of Mn-ZnS quantum dots (QDs) and the specific recognition performance of the aptamer, we built phosphorescent composites from aptamers conjugated with polyethyleneimine quantum dots (PEI-QDs) and applied them to cytochrome c (Cyt c) detection. Specifically, QDs/CBA composites were generated from the electrostatic interaction between the positively-charged PEI-QDs and the negatively-charged Cyt c binding aptamer (CBA). With the presence of Cyt c, the Cyt c can specifically bind with the QDs/CBA composites, and quench the RTP of QDs through photoinduced electron-transfer (PIET). Thereby, an optical biosensor for Cyt c detection was built, which had a detection range of 0.166-9.96 µM and a detection limit of 0.084 µM. This aptamer-mediated phosphorescent sensor with high specificity and operational simplicity can effectively avoid the interference of scattering light from complex substrates. Our findings offer a new clue for building biosensors based on QDs and aptamers.
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BACKGROUNDS: The incidence of nonalcoholic fatty liver disease (NAFLD) is rapidly increasing due to the prevalence of obesity. NAFLD is a major risk factor of hepatocellular carcinoma (HCC). Even with successful surgical removal, the presence of NAFLD is associated with an increased recurrence of HCC. Despite the extensive study of NAFLD, its underlying mechanism(s) remains essentially unknown and there are no FDA-approved drugs for its treatment. Alterations in microRNA (miR) expression have been observed in human fatty livers. However, regulatory mechanism(s) of miRNA biogenesis and their role in regulating the development of NAFLD is poorly described. METHODS: We used immunohistochemistry, luciferase assays and immunoblotting to study the regulatory mechanism of miR-378 biogenesis. Wild-type mice kept on a high fat diet (HFD) were injected with miR-378 inhibitors or a mini-circle expression system containing miR-378 to study loss and gain-of functions of miR-378. RESULTS: miR-378 was significantly increased in fatty livers of dietary obese mice and human hepatoma HepG2 cells with accumulated lipid. Further studies identified NRF1 (Nuclear receptor factor 1), a key regulator of fatty acid oxidation (FAO), as a direct target of miR-378. Overexpression of miR-378 impaired FAO and promoted lipid accumulation in murine hepatoma Hepa1-6 cells. In contrast, knockdown of miR-378 using its ASO (anti-sense oligo) improved FAO and reduced intracellular lipid content in Hepa1-6 cells. Liver-specific expression of miR-378 impaired FAO, which subsequently promoted the development of hepatosteatosis. Antagonizing miR-378 via injecting miR-378-ASO into HFD-treated mice led to increased expression of Nrf1, improved FAO and decreased hepatosteatosis. Additional knockdown of up-regulated Nrf1 offset the effects of miR-378-ASO, suggesting that Nrf1 mediated the inhibitory effect of miR-378-ASO on hepatosteatosis. Furthermore, Nrf1 was identified as a transcriptional repressor of miR-378. Ablation of Nrf1 using its shRNA in livers led to increased miR-378, which subsequently resulted in reduced FAO and elevated hepatic lipid content. CONCLUSIONS: These findings identified a negative feedback loop between miR-378 and Nrf1 that promotes the pathogenesis of hepatosteatosis, and suggests the use of miR-378 as a potential therapeutic target for NAFLD.
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Dieta Alta en Grasa/efectos adversos , Retroalimentación Fisiológica/fisiología , MicroARNs/metabolismo , Enfermedad del Hígado Graso no Alcohólico/metabolismo , Factor Nuclear 1 de Respiración/metabolismo , Obesidad/metabolismo , Animales , Células Hep G2 , Humanos , Metabolismo de los Lípidos , Hígado/metabolismo , Ratones , MicroARNs/genética , Enfermedad del Hígado Graso no Alcohólico/etiología , Enfermedad del Hígado Graso no Alcohólico/genética , Factor Nuclear 1 de Respiración/genética , Obesidad/genéticaRESUMEN
Biosensors based on excellent optical properties of quantum dots (QDs) nanohybrids are efficient for biological detection. In this work, a room-temperature phosphorescent (RTP) PDAD-Mn-ZnS QDs biosensor was constructed with poly(diallyldimethylammonium chloride) (PDAD) as the modifier of MPA-capped Mn-ZnS QDs, and used to detect hyaluronic acid (HA). The newly-added HA induced severe electrostatic interaction with PDAD-Mn-ZnS QDs, leading to the aggregation between PDAD-Mn-ZnS QDs and HA and thereby enhancing RTP. The enhancement of RTP was proportional to the HA concentrations within certain ranges. On this basis, a high-performance HA sensor was built and this sensor had a detection limit of 0.03 µg mL-1 and a detection range of 0.08-2.8 µg mL-1. This proposed RTP sensor can avoid interferences from the background fluorescence or scattering light of the matrix that are encountered in spectrofluorometry. Thus, this biosensor is potentially suitable for detection of HA in real samples without complicated pretreatment.
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In this study, a convenient and sensitive method for the detection of resveratrol was established based on phosphorescence quenching of resveratrol for MPA-capped Mn:ZnS QDs. The quenching intensity was in proportion to the concentration of resveratrol within a certain range. Under optimal conditions, the present assay was valid for detecting resveratrol in the range from 0.03 µM to 14 µM with a detection limit of 0.01 µM. The mechanism of action was also explored, and its application in actual samples was also demonstrated with satisfactory results. This study aims to provide a convenient method for the detection of resveratrol.
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MicroRNAs (miRNAs) play an important role in many biological processes, and its level in plasma and other biological fluids is closely related to many diseases. In this work, a selective room-temperature phosphorescence (RTP) detection method for miRNA was developed based on a duplex-specific nuclease (DSN) -assisted signal amplification strategy and phosphorescence resonance energy transfer (PRET) between poly-diallyldimethylammonium chloride-modified quantum dots (QDs@PDDA) and 6-carboxy-X-rhodamine-modified miRNA sequences complementary oligonucleotide (ROX-ssDNA). The positively charged QDs@PDDA could adsorb negatively charged ROX-ssDNA by electrostatic interaction, whereas the RTP signal of QDs@PDDA could be efficiently quenched by ROX-ssDNA via PRET. In the presence of microRNA-21 (miR-21) and DSN, miR-21 hybridized with ROX-ssDNA initially to form a DNA-RNA heteroduplex as the substrate of DSN, then ssDNA in DNA-RNA heteroduplex would be cleaved into small fragments by DSN and liberate miR-21 to hybridize with another ROX-ssDNA. Eventually, due to weak interaction between ROX-ssDNA fragments and QDs@PDDA, PRET efficiency continually decreased whereas the RTP signal was significantly amplified. By employing the strategy above, quantitative detection of miR-21 in the range of 0.25-40 nM with a detection limit of 0.16 nM was realized, showing excellent performance with simplicity, good selectivity and the ability to be a promising method for miRNA detection.
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Endonucleasas/metabolismo , Transferencia Resonante de Energía de Fluorescencia , MicroARNs/análisis , ADN de Cadena Simple/química , ADN de Cadena Simple/metabolismo , MicroARNs/química , MicroARNs/metabolismo , Hibridación de Ácido Nucleico , Polietilenos/química , Puntos Cuánticos/química , Compuestos de Amonio Cuaternario/química , Rodaminas/química , Electricidad EstáticaRESUMEN
The severe background fluorescence and scattering light of real biological samples or environmental samples largely reduce the sensitivity and accuracy of fluorescence resonance energy transfer sensors based on fluorescent quantum dots (QDs). To solve this problem, we designed a novel target sequence DNA biosensor based on phosphorescent resonance energy transfer (PRET). This sensor relied on Mn-doped ZnS (Mn-ZnS) room-temperature phosphorescence (RTP) QDs/poly-(diallyldimethylammonium chloride) (PDADMAC) nanocomposite (QDs+) as the energy donor and the single-strand DNA-ROX as the energy receptor. Thereby, an RTP biosensor was built and used to quantitatively detect target sequence DNA. This biosensor had a detection limit of 0.16nM and a linear range of 0.5-20nM for target sequence DNA. The dependence on RTP of QDs effectively avoided the interference from background fluorescence and scattering light in biological samples. Moreover, this sensor did not need sample pretreatment. Thus, this sensor compared with FRET is more feasible for quantitative detection of target sequence DNA in biological samples. Interestingly, the QDs+ nanocomposite prolonged the phosphorescence lifetime of Mn-ZnS QDs by 2.6 times to 4.94ms, which was 5-6 magnitude-order larger than that of fluorescent QDs. Thus, this sensor largely improves the optical properties of QDs and permits chemical reactions at a long enough time scale.
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Técnicas Biosensibles/métodos , ADN/aislamiento & purificación , Mediciones Luminiscentes/métodos , Puntos Cuánticos/química , ADN/química , ADN de Cadena Simple/química , Transferencia Resonante de Energía de Fluorescencia/métodos , Límite de Detección , Manganeso/química , Nanocompuestos/química , Sulfuros/química , Compuestos de Zinc/químicaRESUMEN
A QDs-DNA nano-probe was made by combining Mn-doped ZnS room-temperature phosphorescence (RTP) quantum dots (QDs) and DNA. Then an RTP sensor for quantitative detection of genetically-modified mark sequence cauliflower mosaic virus 35S promoter (Ca MV 35S) DNA was built on basis of phosphorescent resonance energy transfer (PRET). The underlying principles were that a QDs-DNA water-soluble nano-probe was built by connecting single-strand DNA to the surfaces of QDs via a ligand exchange method. This probe had good RTP performance and could well identify Ca MV 35S. Thereby, the simple, rapid and efficient detection of genetically-modified organisms was realized. With the increase of target DNA sequence, the phosphorescent intensity of QDs was gradually reduced due to the energy transfer between QDs and the organic quencher BHQ2. This sensor had a detection limit of 4.03nM and a detection range of 12-300nM. Moreover, this sensor had high selectivity. This sensor could effectively detect the target DNA compared with mismatched and random sequences. Thus, this method is very promising for biological analysis.
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Caulimovirus/genética , Sondas de ADN/química , ADN de Cadena Simple/química , ADN Viral/análisis , Mediciones Luminiscentes/métodos , Puntos Cuánticos/química , Técnicas Biosensibles/métodos , Sondas de ADN/genética , ADN de Cadena Simple/genética , ADN Viral/genética , Transferencia de Energía , Regiones Promotoras Genéticas , Puntos Cuánticos/ultraestructura , Sulfuros/química , Transgenes , Compuestos de Zinc/químicaRESUMEN
A simple phosphorescence method is proposed for quercetin detection based on Al(3+)-amplified room-temperature phosphorescence (RTP) signals of 3-mercaptopropionic acid (MPA)-capped Mn-doped ZnS quantum dots (QDs). The sensor was established based on some properties as follows. Al(3+) can interact with carboxyl groups on the surface of MPA-capped Mn-doped ZnS QDs via chelation, which will lead to the aggregation of QDs and amplification of RTP signals, After the addition of quercetin, it can form more stable complex with Al(3+) in alkaline aqueous solution and dissociate Al(3+) from the surface of Mn-doped ZnS QDs, which will result in significant recovery of RTP intensity of the MPA-capped Mn-doped ZnS-Al(3+) system. Under the optimized conditions, the change of RTP intensity was proportional to the concentration of quercetin in the range from 0.1 to 6.0 mg L(-1), with a high correlation coefficient of 0.996 and a detection limit of 0.047 mg L(-1). The proposed method is potentially suitable for detection of quercetin in real samples without complicated pretreatment.
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Ácido 3-Mercaptopropiónico/química , Acetatos/química , Compuestos de Aluminio/química , Antioxidantes/análisis , Indicadores y Reactivos/química , Nitratos/química , Puntos Cuánticos/química , Quercetina/análisis , China , Humanos , Límite de Detección , Mediciones Luminiscentes , Microscopía Electrónica de Transmisión , Tamaño de la Partícula , Puntos Cuánticos/ultraestructura , Quercetina/sangre , Quercetina/orina , Reproducibilidad de los Resultados , Espectrofotometría Ultravioleta , Espectroscopía Infrarroja por Transformada de Fourier , Propiedades de Superficie , Difracción de Rayos X , Sulfato de Zinc/químicaRESUMEN
Quantum dot (QD) nanohybrids provide an effective route to explore the new properties of materials and are increasingly used as highly valuable sensitive (bio) chemical probes. Interestingly, the room-temperature phosphorescence (RTP) of 3-mercaptopropionic acid (MPA)-capped Mn-doped ZnS QDs could be remarkably enhanced by the addition of protamine. Based on the above finding, a simple, sensitive, and selective method for rapid detection of protamine was successfully designed. With this method, protamine as a cationic peptide interacts electrostatically with MPA-capped Mn-doped ZnS QDs to form MPA-capped Mn-doped ZnS QD/protamine complexes, which leads to the aggregation of QDs and enhances the RTP intensity. Under the optimized conditions, the RTP intensity change was linearly proportional to the concentration of protamine in the range 0.2-3.0 µg ml(-1), and the limit of detection was 0.14 µg ml(-1). The proposed method was successfully applied to detect protamine in protamine sulfate injection and human serum samples with satisfactory results, and the recovery ranged from 96.5 to 105.6%.