RESUMEN
This work developed a near-infrared (near-IR) light-activated non-enzymatic signal-off photoelectrochemical (PEC) immunoassay for the ultrasensitive detection of α-fetoprotein (AFP) on the basis of branched polyethylenimine (BPEI)-modified upconversion nanoparticle (UCNP)@CdTe quantum dot (QD) nanostructures by coupling with the synergistic effect of dual-purpose copper ions. Emission light originated from NaYF4:Yb,Er UCNP was directly utilized through the electrostatic bonding of CdTe QDs to excite the separation of electron-hole pairs, resulting in the generation of obvious photocurrent under a 980 nm laser. By using polyclonal antibody-labeled cupric oxide nanoparticle as the secondary antibody, the nanolabel was introduced into the monoclonal anti-AFP antibody-modified microplates in the presence of target AFP. After treatment with acid, the as-released copper ion decreased the photocurrent through the synergistic effect with two issues: one was initially to form coordination with BPEI on the surface of UCNP, and then the near-IR excitation light and upconversion luminescence were attenuated due to the internal filter effect; another was to snatch the electrons flowing from the valence band of CdTe QD as the exciton trapping sites. Under optimal conditions, the dual-purpose Cu2+-activated signal-off PEC immunosensing platform exhibited a dynamic linear range from 10 pg mL-1 to 50 ng mL-1, accompanying the decreasing photocurrent with the increment of AFP concentration at an experimental detection limit of 1.2 pg mL-1. Importantly, good accuracy was achieved by this method in comparison with the results with human AFP ELISA kit for analysis of human serum samples. This dual-purpose Cu2+-activated PEC immunoassay brings a promising, enzyme-free and innovative thinking for the detection of low-abundance biomarkers.
Asunto(s)
Cobre/química , Técnicas Electroquímicas/métodos , Nanoestructuras/química , Procesos Fotoquímicos , Polietileneimina/química , Puntos Cuánticos , alfa-Fetoproteínas/análisis , Técnicas Biosensibles , Compuestos de Cadmio/química , Humanos , Inmunoensayo , Límite de Detección , Telurio/químicaRESUMEN
This work reports a strategy for glutathione-loaded liposome-encoded magnetic beads initiated by palindromic fragment-mediated single-chain amplification (PFMSCA) for high-precision quantification of a low-abundance aminoglycoside antibiotic (kanamycin; Kana) by using In2O3-ZnIn2S4 (IO-ZIS) as a photoactive matrix. In this strategy, a Kana-recognition region, primer-like palindromic fragment, and polymerization/nicking template are reasonably integrated into one oligonucleotide (hairpin HP1) for target recognition, magnetic separation, and target amplification. Upon target Kana introduction, the Kana-aptamer region in HP1 specifically recognizes the Kana and triggers the palindromic tails intramolecular self-hybridization, amplifying a large number of short fragments in the presence of Klenow fragment polymerase and Nt.BbvCI. The as-generated nick fragments act as a linker to introduce the free hairpin HP2-functionalized glutathione-loaded liposomes (HP2-GLL) onto the surface of the hairpin HP3-modified magnetic beads (HP3-MB), constructing liposome-encoded magnetic beads (HP3-MB-nick-HP2-GLL). Following magnetic separation, the detached glutathione-loaded liposomes (GLL) are lysed by treatment with 1% Triton X-100 to release the glutathione within it, which were then detected as an amplified photocurrent at the IO-ZIS-based photoelectrode. Importantly, this method can be readily carried out by using one oligonucleotide to achieve an exponential amplification effect and open new opportunities for advanced development of robust biodetection systems.
Asunto(s)
Técnicas Biosensibles/métodos , ADN de Cadena Simple/química , ADN de Cadena Simple/genética , Secuencias Invertidas Repetidas , Procesos Fotoquímicos , Electroquímica , Glutatión/química , Límite de Detección , Liposomas/química , Imanes/química , Microesferas , Técnicas de Amplificación de Ácido NucleicoRESUMEN
This work presented an innovative and rationally engineered palindromic molecular beacon (PMB) based "Z-scheme" photoelectrochemical (PEC) biosensing protocol for the selective screening of kanamycin (Kana) through DNA hybridization-induced conformational conversion. Interestingly, the ingeniously designed PMB integrated the multifunctional elements including recognition region, primer-like palindromic fragment, and polymerization-nicking template. The cosensitized structures consisted of CdS quantum dot functionalized hairpin DNA2 (QD-HP2) and region-selectively deposited gold nanoparticles onto {001} facets of bismuth oxychloride (BiOCl-Au). Compared with BiOCl-Au alone, the attachment of CdS QDs onto BiOCl-Au (i.e., BiOCl-Au-CdS QDs) exhibited evidently enhanced photocurrent intensity thanks to the synergistic effect of Z-scheme BiOCl-Au-CdS QDs. After incubation with target Kana, Kana-aptamer binding could induce the exposure of PMB region for hairpin DNA1 (HP1). The exposed palindromic tails hybridized with each other (like a molecular machine) to consume the substrates (dNTPs) and fuels (enzyme) for the releasing of numerous nick fragments (Nick). The as-generated nick fragments could specifically hybridize with the complementary region of QD-HP2, thus resulting in decreasing photocurrent because of the increasing spatial distance for electron transfer between two-type photosensitizers. Under optimum conditions, the PMB-based sensing system exhibited satisfying photocurrent responses toward target Kana within the working range from 50 to 5000 fM at a low detection limit of 29 fM. Impressively, the concept of a palindromic fragment-mediated primer-free biosensing strategy offers a new avenue for advanced development of efficient and convenient biodetection systems.
Asunto(s)
Bismuto/química , Compuestos de Cadmio/química , Técnicas Electroquímicas/métodos , Kanamicina/análisis , Nanopartículas del Metal/química , Puntos Cuánticos/química , Sulfuros/química , Animales , Antibacterianos/análisis , Aptámeros de Nucleótidos/química , Aptámeros de Nucleótidos/genética , Técnicas Biosensibles/métodos , ADN/química , ADN/genética , Técnicas Electroquímicas/instrumentación , Electrodos , Contaminación de Alimentos/análisis , Oro/química , Oro/efectos de la radiación , Secuencias Invertidas Repetidas , Luz , Límite de Detección , Nanopartículas del Metal/efectos de la radiación , Leche/química , Hibridación de Ácido Nucleico , Fotoquímica/métodosRESUMEN
An innovative visible light-driven photoelectrochemical (PEC) immunosensing system was reasonably established for the sensitive detection of prostate-specific antigen (PSA) by using perovskite metal oxide@gold nanoparticle heterostructures (BaTiO3/Au) as the photoactive materials. When plasmonic Au nanoparticles were directly decorated on BaTiO3, a several times surface plasmon resonance (SPR) enhancement of photocurrent density was induced via the injection of hot electrons from visible light-excited Au nanoparticles into the conduction band of BaTiO3, and the combination of BaTiO3 and Au nanoparticles was employed as a promising platform for developing a photoelectrochemical bioanalysis. As a proof of concept, PSA had been detected by the BaTiO3/Au nanocomposite-based PEC sensor. To design such an immunoassay protocol, a monoclonal anti-PSA capture antibody (cAb)-coated microplate and glucose oxidase/polyclonal anti-PSA detection antibody-modified gold nanoparticles (GOx-Au NP-dAb) were used as the immunoreaction platform and signal probe, respectively. Upon the addition of target PSA, a sandwiched immunocomplex was formed accompanying the immuno-recognition between the antigen and antibody, and then the carried GOx could oxidize glucose to produce H2O2. The photocurrent of the BaTiO3/Au nanocomposite-functionalized electrode amplified with increasing H2O2 concentration since H2O2 is considered as a good hole scavenger. On the basis of the above-mentioned mechanisms and the optimized conditions, the assembled PEC immunosensor was linear with the logarithm of the PSA concentration in the range of 0.01-40 ng mL-1 with a detection limit of 4.2 pg mL-1. It afforded rapid response, good precision, and high stability and specificity, implying its great promise in photoelectrochemical immunoassays. More generally, this system sets up an ideal PEC immunosensing system based on the BaTiO3/Au nanocomposites and represents an innovative and low-cost "signal-on" assay scheme for the practical quantitative screening of low-abundance proteins.
Asunto(s)
Compuestos de Bario/química , Oro/química , Calicreínas/sangre , Nanopartículas del Metal/química , Antígeno Prostático Específico/sangre , Titanio/química , Anticuerpos Monoclonales/inmunología , Compuestos de Bario/efectos de la radiación , Técnicas Biosensibles/métodos , Técnicas Electroquímicas/instrumentación , Técnicas Electroquímicas/métodos , Electrodos , Glucosa/análisis , Glucosa Oxidasa/química , Oro/efectos de la radiación , Humanos , Peróxido de Hidrógeno/química , Inmunoensayo/métodos , Calicreínas/inmunología , Luz , Límite de Detección , Nanopartículas del Metal/efectos de la radiación , Nanocompuestos/química , Nanocompuestos/efectos de la radiación , Procesos Fotoquímicos , Prueba de Estudio Conceptual , Antígeno Prostático Específico/inmunología , Resonancia por Plasmón de Superficie/métodos , Titanio/efectos de la radiaciónRESUMEN
Aflatoxin B1 (AFB1) pollution is one of the most serious problems for food safety. In this paper, a split-type photoelectrochemical (PEC) immunoassay was designed for sensitive detection of AFB1 in foodstuffs by using amorphous TiO2 with all-inorganic perovskite CsPbBr3 nanocrystals (CsPbBr3/a-TiO2). The a-TiO2 layer not only improved the stability of CsPbBr3 nanocrystals, but also facilitated charge transfer, which resulted in the increasing photocurrent of the nanocomposites. Initially, a competitive-type enzyme immunoreaction was executed on a high-binding microplate between the analyte and alkaline phosphatase (ALP)-labeled AFB1-bovine serum albumin (AFB1-BSA) conjugate. Accompanied by the formation of the immunocomplex, the carried ALP triggered enzymatic hydrolysis to generate ascorbic acid (AA, as an electron donor) for increasing the photocurrent of the CsPbBr3/a-TiO2-modified electrode. Coupling with the competitive enzyme immunoassay, the photocurrent of the modified electrode decreased with the increase of target AFB1 concentration in a dynamic working range from 0.01 ng mL-1 to 15 ng mL-1 with a limit of detection (LOD) of 2.8 pg mL-1 under optimum conditions. Furthermore, the photoelectrochemical immunoassay was also utilized to detect AFB1 in peanut and corn samples, giving acceptable accuracy in comparison with the referenced AFB1 enzyme-linked immunosorbent assay (ELISA) method.
Asunto(s)
Aflatoxina B1/análisis , Contaminación de Alimentos/análisis , Inmunoensayo/métodos , Nanopartículas/química , Aflatoxina B1/inmunología , Fosfatasa Alcalina/química , Animales , Anticuerpos Inmovilizados/inmunología , Arachis/microbiología , Bromuros/química , Bovinos , Cesio/química , Técnicas Electroquímicas/instrumentación , Técnicas Electroquímicas/métodos , Electrodos , Plomo/química , Límite de Detección , Nanocompuestos/química , Prueba de Estudio Conceptual , Albúmina Sérica Bovina/química , Titanio/química , Zea mays/microbiologíaRESUMEN
A marigold-like SiC@MoS2 nanoflower with a unique Z-scheme structure efficiently achieves the overall conversion of gas phase CO2 with H2O (CO2 (g) + 2H2O (g) = CH4 + 2O2) without any sacrificial reagents under visible light (λ ≥ 420 nm) irradiation. The CH4 and O2 evolution are 323 and 621 µL·g-1·h-1, and stable throughout 5 cycle reactions of total 40 h. This work demonstrates a breakthrough in artificial photosynthesis with the Z-scheme 1D heterojunction constructed by combining 2D semiconductor and 3D semiconductor based on the transfer balance of photogenerated electron and hole.
RESUMEN
Pressure-based bioassays incorporating biomolecular recognition with a catalyzed gas-generation reaction have been developed for gas biosensors, but most involve poor sensitivity and are unsuitable for routine use. Herein we design an innovative gas pressure-based biosensing platform for the detection of Kanamycin (Kana) on polyaniline nanowires-functionalized reduced graphene oxide (PANI/rGO) framework by using platinum nanozyme-catalyzed gas generation. The signal was amplified by coupling with catalytic hairpin assembly (CHA) and strand-displacement amplification (SDA). Upon target Kana introduction, the analyte initially triggered a SDA reaction between hairpin DNA1 and hairpin DNA2, and then induced CHA conjugation between magnetic bead-labeled hairpin DNA3 (MB-H3) and platinum nanoparticle-labeled hairpin DNA4 (Pt-H4) to form a three-dimensional network. Numerous platinum nanoparticles (peroxidase-like nanozymes) were carried over with magnetic beads to reduce hydrogen peroxide into oxygen. The as-produced gas compressed PANI/rGO frameworks (modified to polyurethane sponge, used as the piezoelectric materials) in a homemade pressure-tight device, thus causing the increasing current of PANI/rGO sponge thanks to its deformation. The change in the current caused by the as-generated gas pressure was determined on an electrochemical workstation. Under optimum conditions, PANI/rGO sponge exhibited outstanding compressibility, stable signal-waveform output, fast response and recovery time (≈109 ms), and the current increased with the increasing Kana concentration within a dynamic working range of 0.2-50 pM at a detection limit of 0.063 pM. Good reproducibility, specificity, and acceptable precision were acquired for Kana analysis. In addition, the accuracy of this method was monitored to evaluate real milk samples with the well-matched results obtained by using the referenced Kana ELISA kit.
RESUMEN
A novel photoelectrochemical (PEC) enzyme immunoassay was designed for the ultrasensitive detection of alpha-fetoprotein (AFP) based on near-infrared (NIR) light-excited core-core-shell UCNP@Au@CdS upconversion nanospheres. Plasmonic gold (Au) between the sandwiched layers was not only utilized as an energy harvester for the collection of the incident light but also acted as an energy conveyor to transfer the energy from upconversion NaYF4:Yb3+,Er3+ (UCNP) to semiconductor CdS, thus exciting the efficient separation of electron-hole pairs by the generated H2O2 of enzyme immunoreaction under the irradiation of a 980 nm laser. By virtue of high catalytic activity of natural enzymes, gold nanoparticles heavily functionalized with glucose oxidase (GOx) and polyclonal anti-AFP antibody were utilized to generate H2O2. A sandwiched immunoreaction was first carried out in a monoclonal anti-AFP antibody-coated microplate by using an antibody-labeled gold nanoparticle as secondary antibody. Accompanying the gold nanoparticle, the carried GOx oxidized glucose in H2O2, thereby resulting in the enhanced photocurrent via capturing holes on the valence band of CdS to promote the separation of electron-hole pairs. Under optimum conditions, the NIR light-based PEC immunosensing system exhibited good photocurrent responses toward target AFP within the dynamic working range of 0.01-40 ng mL-1 at a detection limit of 5.3 pg mL-1. Moreover, the NIR light-based sensing platform had good reproducibility and high selectivity. Importantly, good well-matched results obtained from NIR light-based PEC immunoassay were acquired for the analysis of human serum specimens by using AFP ELISA kit as the reference.
Asunto(s)
Compuestos de Cadmio/química , Oro/química , Técnicas para Inmunoenzimas/métodos , Sustancias Luminiscentes/química , Nanosferas/química , Sulfuros/química , alfa-Fetoproteínas/análisis , Anticuerpos Inmovilizados/química , Técnicas Biosensibles/métodos , Técnicas Electroquímicas/métodos , Humanos , Rayos Infrarrojos , Límite de Detección , Nanosferas/ultraestructura , Reproducibilidad de los Resultados , Espectrofotometría InfrarrojaRESUMEN
BACKGROUND: Apatinib is an oral anti-angiogenic drug that mainly targets vascular endothelial growth factor receptor 2 (VEGFR-2) and is widely used in a variety of solid tumours. The purpose of this study is to evaluate the clinical efficacy and safety of apatinib in patients with advanced platinum-resistant relapsed epithelial ovarian cancer (EOC). METHODS: A retrospective analysis was performed, the clinical data of patients with stage IIIC-IV platinum-resistant relapsed EOC between January 2014 and May 2018 were collected. The objective response rate (ORR), disease control rate (DCR), progression-free survival (PFS), and overall survival (OS) were reviewed and evaluated. The propensity score matching (PSM) method was used to determine the final case data included in this study. RESULTS: According to 1:2 propensity matching, 108 patients were finally taken into account: 36 in the apatinib group and 72 in the control group. The follow-up ended in January 2019, and the median follow-up time was 28 months. In the apatinib group, ORR was 30.56% and DCR was 66.67%, whereas in the control group, ORR was 16.67% and DCR was 44.44%. In the apatinib group, median PFS was 6.0 months (95% CI 3.69-8.31) and median OS was 15.8 months (95% CI 6.99-24.6), while in the control group, median PFS was 3.3 months (95% CI 2.44-4.16) and median OS was 9.2 months (95% CI 6.3-12.06); the difference was statistically significant (P < 0.05). Apatinib was more effective than conventional chemotherapy in reducing the risk of PFS [HR 0.40 (95% CI 0.22-0.76), P = 0.0017] and OS [HR 0.40 (95% CI 0.21-0.73), P = 0.002]. Multivariate Cox analysis showed that the course of treatment and decrease in serum CA125 levels are independent risk factors for PFS in patients, while apatinib, the length of treatment course and the location of the lesion are independent risk factors for recurrence affecting the OS of patients. The main grade 3-4 adverse events in the apatinib group were hypertension, hand-foot syndrome, and oral mucosal ulcers, and all adverse events were controllable. CONCLUSION: Apatinib was found to be both safe and effective in patients with advanced platinum-resistant relapsed EOC. More in-depth clinical research and applications should be carried out.
Asunto(s)
Antineoplásicos , Neoplasias Ováricas , Femenino , Humanos , Antineoplásicos/farmacología , Antineoplásicos/uso terapéutico , Carcinoma Epitelial de Ovario/tratamiento farmacológico , Neoplasias Ováricas/tratamiento farmacológico , Platino (Metal)/farmacología , Platino (Metal)/uso terapéutico , Estudios Retrospectivos , Factor A de Crecimiento Endotelial Vascular , Resistencia a AntineoplásicosRESUMEN
Electrocatalytic carbon dioxide reduction reaction (CO2 RR) holds significant potential to promote carbon neutrality. However, the selectivity toward multicarbon products in CO2 RR is still too low to meet practical applications. Here the authors report the delicate synthesis of three kinds of Ag-Cu Janus nanostructures with {100} facets (JNS-100) for highly selective tandem electrocatalytic reduction of CO2 to multicarbon products. By controlling the surfactant and reduction kinetics of Cu precursor, the confined growth of Cu with {100} facets on one of the six equal faces of Ag nanocubes is realized. Compared with Cu nanocubes, Ag65 -Cu35 JNS-100 demonstrates much superior selectivity for both ethylene and multicarbon products in CO2 RR at less negative potentials. Density functional theory calculations reveal that the compensating electronic structure and carbon monoxide spillover in Ag65 -Cu35 JNS-100 contribute to the enhanced CO2 RR performance. This study provides an effective strategy to design advanced tandem catalysts toward the extensive application of CO2 RR.
RESUMEN
An all-solid-state metal-mediated Z-scheme photoelectrochemical (PEC) immunoassay was designed for sensitive detection of prostate-specific antigen (PSA) by using WO3-Au-CdS nanocomposite as photoactive material and copper ion (Cu2+) as an inhibitor. The Z-scheme PEC system comprising of CdS nanoparticle (photosystem I; PS I), WO3 nanorod (photosystem II; PS II) and gold nanoparticle (Au NP; solid electron mediator) was reasonably established by a simple and green synthetic method. As an important part of Z-scheme system, the sandwiched gold nanoparticles between electron donor materials and hole provider materials could accelerate electron transfer from positive conduction band (CB) of WO3 to negative valence band (VB) of CdS, thus resulting in high-efficient separation of the carriers. In the presence of target PSA, a sandwiched immunoreaction was executed between capture antibody-coated microplate and CuO nanoparticle-labeled detection antibody. Thereafter, CuO nano labels were dissolved into Cu2+ ions under acidic condition to decrease the photocurrent of Z-scheme WO3-Au-CdS thanks to the formation of exciton trapping center of CuxS (xâ¯=â¯1,2) on the surface. Under optimum conditions, Z-scheme PEC immunoassay exhibited good photocurrents toward target PSA within a linear range of 0.01-50â¯ngâ¯mL-1 at a limit of detection of 1.8â¯pgâ¯mL-1. Moreover, the Z-scheme PEC immunoassay had high selectivity and accuracy. Importantly, this method provides a new horizon for detection of disease-related biomarkers with high sensitivity.
Asunto(s)
Anticuerpos Inmovilizados/química , Técnicas Biosensibles/métodos , Compuestos de Cadmio/química , Cobre/química , Inmunoensayo/métodos , Nanoestructuras/química , Óxidos/química , Antígeno Prostático Específico/sangre , Sulfuros/química , Tungsteno/química , Técnicas Electroquímicas/métodos , Transporte de Electrón , Oro/química , Humanos , Límite de Detección , Nanopartículas/química , Nanotubos/química , Procesos FotoquímicosRESUMEN
Herein a versatile photoelectrochemical (PEC) bioanalysis platform for sensitive and specific screening of low-abundance antibiotics (kanamycin, Kana, used in this case) was innovatively designed using rGO-Bi2WO6-Au as photoactive matrix and target-induced branched hybridization chain reaction (t-bHCR) for efficient signal amplification. To realize the high-performance of our PEC bioanalysis system, rational introduction of reduced graphene oxide (rGO) and Au nanoparticles (Au NPs) greatly accelerated the electron transfer and enhances photoactivity. As expected, the ternary nanocomposite (i.e., rGO-Bi2WO6-Au) system with cascade energy level exhibited intense PEC signal responses thanks to multistep electron-transfer (MET) mechanism. Upon sensing the target Kana, t-bHCR is readily implemented, thus resulting in the assembly of numerous CuS nanoparticle (CuS NP). As a result, the loading CuS NPs from hyper-branched structure boosted the electron donors (ascorbic acid) consumption and enhanced the steric hindrance, synergistically decrease the photoelectric response. Under the optimized testing conditions, the t-bHCR-based PEC bioanalysis exhibited superior analytical performance with a linear range of 1â¯pM to 5â¯nM target Kana and limit of detection down to 0.78â¯pM. Additionally, favorable stability, great anti-interference ability and satisfactory accuracy for the analysis of actual samples were acquired. Impressively, the concept of t-bHCR-mediated provides an alternative to construct PEC bioanalysis and inspire more interest in the design of advanced PEC bioanalysis through nucleic acid-related signal amplification.
Asunto(s)
Técnicas Biosensibles , Técnicas Electroquímicas , Grafito/química , Anticuerpos Inmovilizados/química , Bismuto/química , Oro/química , Humanos , Límite de Detección , Nanopartículas del Metal/química , Nanocompuestos/química , Hibridación de Ácido Nucleico , Óxidos/química , Antígeno Prostático Específico , Compuestos de Tungsteno/químicaRESUMEN
This work reports the proof-of-concept of an ultrasensitive label-free electrochemical aptasensor for Kanamycin (Kana) detection coupling strand-displacement amplification (SDA) with hybridization chain reaction (HCR). In the presence of target Kana, the analyte triggers conformational change of hairpin HP1 (HP1) and two-staged SDA to produce short single-stranded DNA (S1) with the aid of KF polymerase and nicking endonuclease. Meanwhile, the as-produced S1 hybridizes with the immobilized hairpin HP2 (HP2) on the electrode to open the hairpin, thereby resulting in the formation of DNA duplex. Thereafter, DNA duplex is selectively digested by Exo III accompanying S1 recycling. The residual single-stranded probe (S2) on the electrode opens another two hairpins in sequence and propagates a chain reaction of hybridization events between two alternating hairpins (H1 and H2) to form a long nicked double-helix. Upon addition of redox-active methylene blue (MB), numerous indicators are intercalated into the grooves of double-helix DNA polymers, each of which produces an electrochemical signal within the applied potentials. Under optimum conditions, the SDA/HCR-based electrochemical aptasensor exhibits a high sensitivity for detection of Kana down to 36â¯fM with a linear range from 0.05 to 200 pM. Additionally, the as-prepared aptasensor is successfully employed to determinate the Kana in animal derived food (milk). With the advantages of high sensitivity, label-free strategy and excellent selectivity, the developed aptasensor possesses great potential application value in food-safety analysis field.
Asunto(s)
Aptámeros de Nucleótidos/química , Técnicas Electroquímicas , Kanamicina/análisis , Leche/química , Técnicas de Amplificación de Ácido Nucleico , Animales , BovinosRESUMEN
In this study, we designed a novel dual-readout biosensing protocol for quantitative or qualitative screening of antibiotic residues (Kanamycin; Kana used in this case) using a spectrofluorometer and via naked-eye detection. This assay comprises two sequential reactions: target-aptamer binding to trigger Exonuclease I (Exo I)-assisted target recycling and a three-way junction-assisted hybridization chain reaction (3WJ-HCR). Gold nanocluster-functionalized manganese dioxide nanosheets (AuNCs-MnO2) were synthesized via a one-pot biomimetic mineralization process and used as signal-generation tags because of the quenching efficiency of MnO2 toward the AuNC's fluorescence. Upon addition of Kana analyte, target-assisted recycling and the 3WJ-HCR reaction were readily implemented on functional magnetic beads, thus resulting in the assembly of numerous alkaline phosphatase (ALP) molecules through the avidin-biotin interaction, which hydrolyzed ascorbic acid 2-phosphate (AAP) to ascorbic acid (AA). The as-prepared AA etched the MnO2 nanosheets into Mn2+ to release the carried AuNCs, thereby recovering the fluorescence of the AuNCs. Meanwhile, visual detection could be performed according to the change in the color of AuNCs-MnO2. Under optimum operating conditions, the intensity of fluorescence increased with the increase in Kana within a dynamic working range from 0.002 nM to 5 nM, with a limit of detection (LOD) of 1.2 pM. A cutoff value of 50 pM Kana could also be obtained in the visual assay on the basis of change in the color from brown to white. In addition, the precision, reproducibility, and selectivity of this method were acceptable. For the analysis of real milk samples with Kana, well-matched results were acquired between the developed fluorescence assay and the referenced Kana enzyme-linked immunosorbent assay (ELISA) kit.
RESUMEN
Acute lung injury (ALI) is a common emergency and severe case in clinic. High mobility group protein box 1 (HMGB1) can be treated as a new anti-inflammatory treatment target. Toll-like receptor 4 (TLR4) is an important receptor of HMGB1. Ketamine is a widely used intravenous anesthetic with good anti-inflammatory and immune regulating function. Whether it can protect ALI through inhibiting HMGB1 and TLR4 expression in lung tissue still needs further investigation. Male SD rats were randomly divided into control, lipopolysaccharide (LPS) group and ketamine intervention group with 15 rats in each group. The rats were euthanatized at 24 h after modeling and the bronchoalveolar lavage fluid (BALF) was collected for HMGB1 and TLR4 level detection. Western Blot was applied to analyze HMGB1 and TLR4 protein expression in the lung tissue. HMGB1 and TLR4 concentration in BALF were 5.369 ± 1.564 ng/ml and 43.980 ± 7.524 pg/ml in the control, respectively. They were 12.358 ± 4.681 ng/ml and 102.538 ± 8.412 pg/ml in LPS group, and 7.399 ± 2.346 ng/ml and 87.208 ± 7.558 pg/ml in ketamine intervention group, respectively. Their levels increased significantly in LPS group and down-regulated after ketamine intervention. HMGB1 and TLR4 protein expression in lung tissue elevated obviously in LPS group, and decreased after ketamine treatment. HMGB1 and TLR4 protein level showed positive correlation in lung tissue (r = 0.921, P < 0.001). Ketamine can inhibit HMGB1 and TLR4 expression in ALI, and alleviate LPS induced rat lung injury.