ACS Applied Materials & Interfaces
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2019
·
IF 8.456
Tianqi Nie,Zhiyu He,Yang Zhou,Jinchang Zhu,Kuntao Chen,Lixin Liu,Kam W. Leong,Hai-Quan Mao,Yongming Chen
Oral delivery of nucleic acid therapy is a promising strategy in treating various diseases because of its higher patient compliance and therapeutic efficiency compared to parenteral routes of administration. However, its success has been limited by the low transfection efficiency resulting from nucleic acid entrapment in the mucus layer and epithelial barrier of the gastrointestinal (GI) tract. Herein, we describe an approach to overcome this phenomenon and improve oral DNA delivery in the context of treating type II diabetes (T2D). Linear PEI (lPEI) was used as a carrier to form complexes with plasmid DNA encoding glucagon-like peptide 1 (GLP-1), a common target in T2D treatments. These nanoparticles were then coated with a mixture of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and 1,2-dimyristoyl-rac-glycero-3-methoxy poly(ethylene glycol)-2000 (DMG-PEG) to render the nanoparticle surface hydrophilic and electrostatically neutral. The surface-modified lPEI/DNA nanoparticles showed higher diffusivity and transport in the mucus layer of the GI tract and mediated high levels of transfection efficiency in vitro and in vivo. Moreover, these modified nanoparticles demonstrated high levels of GLP-1 expression for more than 24 h in the liver, lungs, and intestine in a T2D murine model after a single dose, as well as controlled blood glucose levels within a normal range for at least 18 h with repeatable therapeutic effects upon multiple dosages. Taken together, this work demonstrates the feasibility of an oral plasmid DNA delivery approach in the treatment of T2D through a facile surface modification to improve the mucus permeability and delivery efficiency of the nanoparticles.
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ACS Applied Materials & Interfaces
·
2019
·
IF 8.456
Ding Wang,Yaqin Chai,Yali Yuan,Ruo Yuan
In this work, the array arrangement of cascade enzymes was implemented by alternately and equidistantly anchoring two model enzymes glucose oxidase (GOx) and horseradish peroxidase (HRP) to the vertexes of rigid DNA tetrahedron units in lattice-like nucleic acid scaffold, in which the distance between any adjacent cascade enzymes had been regulated to the optimum for obtaining high enzyme cascade catalytic efficiency. Compared to the enzyme cascade system with no-array arrangement of cascade enzymes, the proposed enzyme cascade system allowed the intermediate H2O2 produced by GOx catalyzing substrate glucose to concurrently and equidistantly diffuse toward the four adjacent HRP enzyme surfaces. In this case, the invalid diffusion effect of intermediate H2O2 between cascade enzymes could be effectively avoided, thereby promoting the enzyme cascade reaction with high catalytic efficiency. The specific catalytic efficiency (kcat/Km) of the cascade enzyme system with array arrangement had been evaluated, which exhibited catalytic efficiency about 3.6 times higher than that of the randomly arranged cascade enzyme system. As a result, this strategy provided a new avenue for constructing a highly efficient enzyme cascade system with ultimate applications in biosynthesis, bioanalysis, and biodiagnostics.
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ACS Applied Materials & Interfaces
·
2019
·
IF 8.456
Ting Bao,Ruobing Fu,Wei Wen,Xiuhua Zhang,Shengfu Wang
In this paper, a label-free and stimuli-responsive electrochemical biosensing platform was fabricated based on target-driven load release from DNA-gated metal–organic frameworks (MOFs) with cascade amplification. By using MOFs (UiO-66-NH2) as a nanocarrier of electroactive molecules (methylene blue; MB) and the programmably assembled DNA acted as the gatekeeper, the biofunctionalized MOFs (MB@DNA/MOFs) were not only used as an amplified signal label but also worked as three-dimensional tracks for biosensing. In the presence of a target, the nicking endonuclease cleavage process was triggered, leading to the generation of two strands (S1 and S2). Both S1 and S2 act as stimuli to participate in the strand displacement reaction on the MB@DNA/MOFs, which caused the unlocking of the pore to release MB, resulting in the decrease of the signal. Using carcinoembryonic antigen (CEA) as a model target, the cascade-amplified biosensor presented good performance for CEA detection, ranging from 50 fg/mL to 10 ng/mL with a detection limit of 16 fg/mL. The stimuli-responsive DNA-gated MOF-based electrochemical platform exhibited three-dimensional biosensing tracks with rational utilization of the cascade amplification, providing an effective method for cancer biomarker detection.
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Oxidative Medicine and Cellular Longevity
·
2019
·
IF 4.868
Lifang Sun,Min Zhu,Wei Feng,Yiping Lin,Jia Yin,Juan Jin,Yunguang Wang
Idiopathic pulmonary fibrosis (IPF) is a prototype of chronic, progressive, and fibrotic lung disease with high morbidity and high mortality. Menstrual blood-derived stem cells (MenSCs) have proven to be an attractive tool for the treatment of acute lung injury and fibrosis-related diseases through immunosuppression and antifibrosis. However, whether MenSC-derived exosomes have the similar function on pulmonary fibrosis remains unclear. In the present study, exosomes secreted from MenSCs (MenSCs-Exo) were verified by transmission electron microscope (TEM), nanoparticle tracking analyzer (NTA), and western blotting. And MenSC-Exo addition significantly improved BLM-induced lung fibrosis and alveolar epithelial cell damage in mice, mainly reflected in BLM-mediated enhancement of the fibrosis score, blue collagen deposition, dry/wet gravity ratio, hydroxyproline and malondialdehyde levels, and downregulation of glutathione peroxidase, which were all robustly reversed by MenSC-Exo management. Additionally, BLM- and TGF-β1-evoked cellular reactive oxygen species (ROS), mitochondrial DNA (mtDNA) damage, and cell apoptosis were rescued by MenSCs-Exo in vivo and in vitro. Further study indicated that the MenSCs-Exo could transport miRNA Let-7 into recipient alveolar epithelial cells. Let-7 inhibitor administration significantly blocked the exosome-mediated improvement role on lung fibrosis in mice. Mechanistically, Let-7 was able to regulate the expression of lectin-like oxidized low-density lipoprotein receptor-1 (LOX1) through binding to its 3-UTR region. Forced expression of LOX1 promoted the expression of apoptosis-related protein and mtDNA damage markers via regulating NLRP3 which was also confirmed in BLM model mice under the combination therapy of the exosome and Let-7 inhibitor. Collectively, this study demonstrates that exosomal Let-7 from MenSCs remits pulmonary fibrosis through regulating ROS, mtDNA damage, and NLRP3 inflammasome activation. This provides a new approach of exocytosis on the treatment of fibrotic lung disease.
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ANALYTICAL CHEMISTRY
·
2019
·
IF 6.35
Bing Wang,Suwen Shi,Xiaolin Yang,Yue Wang,Honglan Qi,Qiang Gao,Chengxiao Zhang
A separation-free electrogenerated chemiluminescence (ECL) immunoassay for biomarkers has been developed incorporating target assistant proximity hybridization and dynamically competitive hybridization of a DNA ECL signal probe for the first time. In this work, the biomarkers of acute myocardial infarction including cardiac troponin I (cTnI), cardiac troponin T (cTnT), and myoglobin (Myo) were chosen as the model proteins while the corresponding antibody was utilized as a recognition probe and the DNA5 tagged with ruthenium complex was chosen as an ECL signal probe (DNA5-Ru1). The biosensors were fabricated by covalently coupling the capture probe DNA1 onto the surface of a glassy carbon electrode, and then, a competitor ss-DNA2 was hybridized with DNA1. When the biosensor was incubated in the solution containing a target protein, the recognition probes (DNA3-Ab1 and DNA4-Ab2), DNA5-Ru1, and the coreactant tri-n-propylamine, the target protein was bounded with two antibodies of the recognition probes and thus induced the sufficient proximity hybridization of DNA3 with DNA1, DNA4 with DNA5-Ru1, and DNA5-Ru1 with DNA1 and the unwinding of the competitor DNA2 with DNA1, and ECL measurement was performed in separation-free format. It was found that the hybridization base number and length of DNA1 and a competing hybridization of DNA5-Ru1 with DNA2 for DNA1 have important effects. The developed ECL method showed a quite low detection limit of 0.4 pg/mL for cTnI, 0.5 pg/mL for cTnT, and 0.5 ng/mL for Myo. The fabricated biosensor exhibited stability and reusability. This work demonstrated that the developed ECL immunoassay is a promising separation-free and flexible strategy for quantitation of multiple proteins using one biosensor.
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MICROCHIMICA ACTA
·
2018
·Vol. 185
·期 5
·1-9
·
IF 5.705
Huang Jianyong,Li Fenglan,Guo Rubin,Chen Yuyuan,Wang Zhenzhen,Zhao Chengfei,Zheng Yanjie,Weng Shaohuang,Lin Xinhua
Amino-modified carbon dots (C-dots) with positively charged surface were prepared. They display strong blue fluorescence and are shown to act as quenchers of the green fluorescence of FAM-labeled ssDNA such as the F-probe used in this work that was immobilized on the C-dots. On the addition of highly negatively charged heparin (Hep), it will interact with the C-dots and displace the F-probe from C-dots. Once the F-probe is displaced by Hep, its green fluorescence is restored. The intrinsic blue fluorescence of the C-dots remains stable after addition of Hep. Thus, a signal-on ratiometric fluorometric assay was developed for the ultra-sensitive detection of Hep. The underlying mechanisms of quenching and recovery are discussed. Under optimized conditions, the recovery of the ratiometric fluorescence of the system composed of C-dots and quenched F-probe is proportional to the Hep concentration in the range of 0.01–2.0 μg·mL −1 (= 0.00125–0.25 U·mL −1 ). The method was successfully applied to the determination of Hep in spiked serum samples.
Schematic of a signal-on ratiometric fluorometric method for the ultra-sensitive detection of heparin on the basis of the displacement and fluorescence enhancement of adsorbed FAM-labeled ssDNA from amino-modified carbon dots (C-dots) by heparin.
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Journal of Materials Chemistry B
·
2018
·Vol. 6
·期 36
·5718-5728
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IF 4.776
Jiayi Song,Hao Shen,Ye Yang,Zixin Zhou,Ping Su,Yi Yang
The overall stability and activity of immobilized enzyme systems have suffered from non-specific adsorption immobilization of enzymes at the surface of carriers through electrostatic and hydrophobic interactions. Elimination of these non-specific adsorptions of enzymes on the surface of carriers is critical for enzyme reactions. Herein, for the first time, we have prepared zwitterion-functionalized magnetic particles with amino, phosphonate, and thiol functional groups to coimmobilize glucose oxidase and horseradish peroxidase by DNA directed immobilization. The zwitterionic surface of the multifunctional magnetic particles could efficiently suppress non-specific adsorption of different kinds of enzymes, and the immobilized multienzyme catalyst without non-specifically adsorbed enzymes exhibited excellent enzymatic activity, stability, and reusability compared with those of free and non-specifically adsorbed enzymes. The immobilized multienzymes maintained 87% of their enzymatic activity after two weeks of storage at 4 °C and 58% of their enzymatic activity after 90 min of thermal incubation at 60 °C. Furthermore, the immobilized multienzymes exhibited more than 87% enzymatic activity after reuse for 10 cycles. The apparent Km and catalytic efficiency (kcat/Km) values of the immobilized multienzymes were 12.6 mM and 4.03 s−1 mM−1, respectively, which were 0.2- and 11.8-fold better than those of the free enzymes, indicating effective cascade efficiency and substrate affinity. The optimal ratio of GOx and HRP was 1 : 5, and the prepared immobilized enzymes could detect low concentrations of glucose (0.5 μM) with excellent selectivity. Therefore, we believe that the strategy developed in this study can be widely applied in biotechnology, industrial catalysis, and biomedical engineering.
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BIOSENSORS & BIOELECTRONICS
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2018
·
IF 8.173
Chengji Sui,Tingting Wang,Yunlei Zhou,Huanshun Yin,Xiangjian Meng,Shenran Zhang,Geoffrey I.N. Waterhouse,Qin Xu,Yuping Zhuge,Shiyun Ai
5-Hydroxymethylcytosine (5hmC) plays an important role in switching genes on and off in mammals, and it is implicated in both embryonic development and cancer progression. Herein, a novel photoelectrochemical (PEC) biosensor was developed for 5hmC detection based on WS 2 nanosheets as the photoactive material and boronic acid functionalized carbon dots (B-CDs) for signal amplification unit. This biosensor can also be used for T4-β-glucosyltransferase (β-GT) activity assessment. Firstly, WS 2 nanosheets and gold nanoparticles (AuNPs) were immobilized on an ITO electrode surface . Then probe DNA was immobilized on this electrode surface via Au-S bond. Afterwards, the complementary DNA containing 5hmC was then captured on the modified electrode surface by hybridization. Subsequently, β-GT transferred glucose from uridine diphosphoglucose to the hydroxyl groups of the 5hmC residues. After glycosylation , B-CDs could further be immobilized on the modified electrode surface resulting in a strong photocurrent . The PEC biosensor afforded high selectivity , excellent sensitivity and good reproducibility, with detection limits of 0.0034 nM and 0.028 unit/mL for 5hmC and β-GT, respectively. Results demonstrate that the photoelectrochemical strategy introduced here based on WS 2 nanosheets and B-CDs offers a versatile platform for hydroxymethylated DNA detection, β-GT activity assessment and β-GT inhibitor screening.
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RSC Advances
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2018
·Vol. 8
·期 45
·25611-25616
·
IF 2.936
Dan Han,Chunying Wei
In this paper, we construct a fluorescence nucleic acid probe based on DNA-templated silver nanoclusters (DNA-Ag NCs) for the detection of the p53 gene. The fluorescence biosensing of the “turn-on” model is successfully implemented as a result of the target-triggered configurational change in the hairpin DNA probe and the synthesis of fluorescent Ag NCs. With this biosensor, the limit of detection (LOD) for the p53 gene is 3.57 nM and the linear range is 250–2500 nM in phosphate buffer solution, while a LOD of 6.06 nM in the linear range of 250–2500 nM is obtained in 1% diluted fetal calf serum. So this probe, with its advantages of specificity, practical application, easy operation and low cost, will have favourable development prospects in biological sensing and imaging.
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MICROCHIMICA ACTA
·
2019
·Vol. 186
·期 7
·1-7
·
IF 5.479
Xu Chengnan,Ying Yibin,Ping Jianfeng
The authors describe a colorimetric method for determination of kanamycin by using gold nanoparticles (AuNPs) as the element of signal-conversion and by applying hybridization chain reaction-assisted signal amplification. The assay is carried out by monitoring the absorbance change and color change adding salt to the reaction solution containing kanamycin (analyte), hairpin DNA probe, and AuNPs. Three hairpin DNA probes with sticky ends were absorbed on the AuNPs via their sticky ends. Cating with DNA prevents them from salt-induced aggregation (which leads to a color change from red to blue) in the complete absence of kanamycin. In contrast, in the presence of kanamycin, the aptamer hairpin DNA probe binds kanamycin, and the newly exposed section of DNA triggers a cascade of hybridization chain reactions with formation of numerous dsDNAs. On addition of salt, the AuNPs form blue aggregates due to the repulsion between dsDNA and AuNPs. Under optimal conditions, the ration of absorbance at 520 and 630 nm drops with the kanamycin concentration in the range from 1 to 40 μM, and the limit of detection is 0.68 μM. The assay can selectively distinguish kanamycin from other antibiotics. The method was applied to kanamycin detection in (spiked) milk samples and gave excellent recoveries.
Schematic presentation of colorimetric method for kanamycin detection using gold nanoparticles modified with hairpin DNA probes and hybridization chain reaction-assisted amplification.
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SENSORS AND ACTUATORS B-CHEMICAL
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2018
·
IF 5.667
Jiao Wang,Ziying Zhang,Xia Gao,Xiaodong Lin,Yaqing Liu,Shuo Wang
Lead ion (Pb 2+ ) is known as a major non-degradable environmental pollutant and is quite harmful to human health. In our investigation, a novel ratiometric fluorescent nanosensor with DNA-AgNCs as single fluorophore has been successfully constructed for ultrasensitive and specific detection of Pb 2+ . The single-strand DNA templated silver nanoclusters present green emission, which can be converted into red emission after approaching a specific DNA segment by forming duplex with its complementary DNA strands (ds-DNA-AgNCs). The ds-DNA-AgNCs contains rA cleavage site of Pb 2+ -dependent DNAzyme configuration. In the presence of Pb 2+ , the specific DNA segment would be released from the ds-DNA-AgNCs, resulting in discoloration of DNA-AgNCs from red to green. Since Pb 2+ can successively cleave the rA site, the signal change from red to green emission is amplified. Thus, benefiting from the Pb 2+ -dependent DNAzyme and the charming properties of DNA-AgNCs, the proposed platform exhibits a good linear relationship from 0.001 nM to 10 nM with a detection limit of 1.0 pM for Pb 2+ determination, which is lower than most of the reported Pb 2+ biosensors. Moreover, this sensitive ratiometric fluorescent probe exhibits excellent selectivity toward Pb 2+ detection in real samples, such as lake water, tap water, and human serum samples, illustrating the huge potential applications for complicated samples in the future.
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ANALYTICAL AND BIOANALYTICAL CHEMISTRY
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2013
·Vol. 405
·期 8
·2435-2442
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IF 3.659
Wang Xiaoli,Wang Hailin
A variety of environmental pollutants may cause abnormal DNA methylation, which further disturb gene expression. In this work, we developed a rapid and sensitive method for the characterization and identification of the epigenotoxicity of environmental pollutants in terms of DNA methylation. The method combines in vitro inhibition reactions of a model DNA methyltransferase (DNMT) with rapid and sensitive capillary electrophoresis–laser-induced fluorescence (CE-LIF) immunoassays. This method was first assessed using two known DNMT inhibitors, (–)-epigallocatechin-3-gallate and RG108, and then applied to epigenotoxic evaluation of four aldehydes and six benzo-1,4-quinones. It was found that all these electrophilic chemicals could inhibit DNMT activity, probably due to their interactions with the active sites of DNMT. Interestingly, benzo-1,4-quinones displayed more inhibitory effects on DNMT activity than aldehydes. Among the tested six benzo-1,4-quinones, halogenated benzo-1,4-quinone showed higher inhibitory activity than non-halogenated p -benzo-1,4-quinone. Owing to its speed and sensitivity, our method will be potentially applicable for fast epigenotoxic screening of environmental pollutants and mechanistic study of environmental epigenetics.
Epigenotoxicity evaluation of environmental pollutants
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