Biosensors-Basel
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2023
·Vol. 13
·期 8
·764
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IF 5.4
Ruiting Chen,Hejing Chen,Huaping Peng,Yanjie Zheng,Zhen Lin,Xinhua Lin
Drug resistance in cancer is associated with overexpression of the multidrug resistance (MDR1) gene, leading to the failure of cancer chemotherapy treatment. Therefore, the establishment of an effective method for the detection of the MDR1 gene is extremely crucial in cancer clinical therapy. Here, we report a novel DNA biosensor based on an aligned multi-walled carbon nanotube (MWCNT) array modified electrode with 3D nanostructure for the determination of the MDR1 gene. The microstructure of the modified electrode was observed by an atomic force microscope (AFM), which demonstrated that the electrode interface was arranged in orderly needle-shaped protrusion arrays. The electrochemical properties of the biosensor were characterized by cyclic voltammetry (CV), differential pulse voltammetry (DPV), and electrochemical impedance spectroscopy (EIS). Chronocoulometry (CC) was used for the quantitative detection of the MDR1 gene. Taking advantage of the good conductivity and large electrode area of the MWCNT arrays, this electrochemical DNA sensor achieved a dynamic range from 1.0 × 10−12M to 1.0 × 10−8M with a minimal detection limit of 6.4 × 10−13M. In addition, this proposed DNA biosensor exhibited high sensitivity, selectivity, and stability, which may be useful for the trace analysis of the MDR1 gene in complex samples.Keywords:multidrug resistant gene;multi-walled carbon nanotube array;electrochemical DNA biosensor;3D sensing interface
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JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
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2023
·
IF 3.4
Tong Li,Fei-Fei Guo,Xi-Ping Mu,Ping Sun,Xue Zhang,Zhi-Hao Xu,Ri-Lei Yu,Ya-Mu Xia,Wei-Wei Gao
In this work, novel potential anthraquinone-temozolomide (TMZ) antitumor hybrids N -(2-((9,10-dioxo-9,10-dihydroanthracen-1-yl)amino)ethyl)−3-methyl-4-oxo-3,4-dihydroimidazo [5, 1-d][1,2,3,5]tetrazine-8-carboxamide ( C-1) and 2-(9,10-dioxo-9,10-dihydroanthracen-1-yl)amino) ethyl-3-methyl-4-oxo-3,4-dihydroimidazo[5,1- d ][1,2,3,5]tetrazine-8-carboxylate ( C - 9 ) were designed and synthesized successfully. The electrochemical behaviors of C - 1 ( C - 9 ) involved the reversible processes of 9,10-anthraquinone ring, the irreversible reduction and oxidation processes of TMZ ring. Electrochemical biosensors were constructed with ct DNA, poly (dG) and poly (dA) modifying the surface of glassy carbon electrode (GCE) to evaluate the DNA oxidative damage caused by the interaction of C - 1 ( C - 9 ) with DNA. Anthracycline skeleton and TMZ ring in C - 1 ( C - 9 ) could exhibit bifunctional effects with both intercalating and alkylation modes toward DNA strands. The DNA biosensor had good practicability in mouse serum. The results of gel electrophoresis further demonstrated that C - 1 ( C - 9 ) could effectively intercalated into ct DNA and disrupt plasmid conformation. Finally, anthraquinone-TMZ hybrid C-1 possessed high cytotoxicity toward A549 and GL261 cells, which could be a novel and optimal candidate for the clinic antitumor treatment.
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ANALYTICAL CHEMISTRY
·
2021
·
IF 6.986
Xiaoqing Wu,Cheng Che,Xinmeng Wang,Qiujiao Du,Huageng Liang,Pengcheng Gao,Fan Xia
DNA species are recognized as a powerful probe for nanochannel analyses to address the issues of specific target recognition and highly efficient signal conversion due to their programmable and predictable Watson–Crick bases. However, in the conventional view, abundant sophisticated DNA structures synthesized by DNA amplification strategies are unsuitable for use in nanochannel analyses owing to their low probability to enter a nanochannel restricted by the smaller opening of the nanochannel, as well as the faint ion signal produced by the steric effect. Here, we present an integrated strategy of nanochannel analyses that combines the target recognitions by encoded rolling circle amplification (RCA) in solution and the ionic signal enhancement by the space charge effect through the immobilization of highly negative-charged RCA amplicons on the outer surface of the nanochannels. Owing to the highly negative-charged RCA amplicons with 100 nm sizes, a sharp increase of ionic current up to 7454% has been achieved. The RCA amplicon triggered by mRNA-21 on the outer surface of the poly(ethylene terephthalate) membrane with a single nanochannel realized the single-base mismatch detection of mRNA-21 with a sensitivity of 6 fM. The DNA amplicon endows the nanochannel with high sensitivity and selectivity that could extend to other applications, such as DNA sequencing, desalination, sieving, and water–energy nexus.
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ANALYST
·
2023
·
IF 4.2
Hongding Zhang,Yinhui Su,Jiamiao Zhao,Huixi Song,Xiaohong Zhou
The accurate and sensitive quantification of DNA methylation is significant for the early diagnosis of cancer. In this work, an alkaline phosphatase (ALP) triggered in situ fluorogenic reaction between ascorbic acid (AA) and 2,3-DAN was employed as a ratiometric fluorescent probe for the accurate and sensitive detection of DNA methylation with the assistance of ALP encapsulated liposomes. The quinoxaline derivative with a yellow fluorescence emission (I525) was generated from the reaction between AA and 2,3-DAN. Meanwhile, the consumption of 2,3-DAN declined its fluorescence intensity (I386). A ratiometric fluorescent probe (I525/I386) constructed by the above in situ fluorogenic reaction was applied for the accurate detection of DNA methylation. The methylated DNA was first captured by its complementary DNA in 96-well plates. Then, 5mC antibody (Ab) linked liposomes that were encapsulated with ALP recognized and combined with the methylation sites of the target DNA. After the liposomes were lysed by Triton X-100, the released ALP triggered the hydrolysis of ascorbic acid diphosphate (AAP) to form AA, participating in the fluorogenic reaction with 2,3-DAN to produce a quinoxaline derivative. Thus, the ratiometric fluorescence detection of DNA methylation was achieved using I525/I386 values. Using the ALP-enzyme catalyzed reaction and liposomes as signal amplifiers, a low detection limit of 82 fM was obtained for DNA methylation detection. Moreover, the accuracy of the assay could be improved using ratiometric fluorescent probes. We hope that the proposed assay will pave a new way for the accurate determination of low-abundance biomarkers.
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TALANTA
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2023
·
IF 6.1
Hongding Zhang,Sifei Wu,Hui-Jin Xiao,Hai-Bo Wang,Linxia Fang,Jun-Tao Cao
Ultrasensitive analytical methods are still urgent for the discovery of trace level biomarkers and the early clinical diagnosis of disease. In this work, an ultrasensitive universal sensing platform was constructed by integrating fluorescent assay with chemical-chemical redox cycling signal amplification strategy. Using Ru@SiO 2 nanoparticles wrapped by MnO 2 nanosheets (Ru@SiO 2 @MnO 2 ) as fluorescent probe , the chemical-chemical redox cycling system was conducted upon ascorbic acid (AA) and tris(2-carboxyethyl)phosphine (TCEP) as reductants and MnO 2 nanosheets as oxidant. The MnO 2 nanosheets not only could quench the fluorescence of Ru@SiO 2 nanoparticles to reduce the background, but also could serve as oxidants to react with AA, generating dehydroascorbic acid (DHA). The DHA was reduced by TCEP in turn to form AA that participated in the next cycling of chemical-chemical redox reaction. Thus, the constantly released AA from the chemical-chemical redox cycling system could massively etch MnO 2 nanosheets on Ru@SiO 2 surface, making the fluorescence of Ru@SiO 2 nanoparticles greatly recovered. It was shown that the sensitivity of the fluorescent assay was improved almost 52 times by utilizing the chemical-chemical redox cycling signal amplification strategy. This strategy was further employed to detect DNA methylation with the aid of AA-encapsulated liposomes that were modified with 5 mC antibodies to bind with the methylated DNA captured in 96-well plate. A detection of limit down to 16.2 fM was achieved for the detection of methylated DNA. It's believed that the incorporation of chemical-chemical redox cycling signal amplification strategy into fluorescent sensing paves a new way for ultrasensitive detection of biomarkers.
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INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2023
·
IF 8.2
Jiadong Pang,Baoxiong Zhuang,Li-Ming Zhang
Pancreatic cancer is one of the most lethal malignancies in the world and remains one of the leading causes of cancer related death. For its treatment, a lot of investigations have dealt not only with individual chemotherapy by using polymeric carriers to deliver anticancer drugs, but also with individual gene therapy by using polymeric carriers to deliver nucleic acids such as small interfering RNA (siRNA) and plasmid DNA . However, relatively few studies have been focused on the co -delivery of gene and anticancer drug by multifunctional polymeric carriers for its synergistic therapy. In this work, a DPLL-functionalized amylose (ADP) was prepared by the click reaction between azidized amylose and propargyl focal point poly( l -lysine) dendrons, and then used to co-deliver plasmid pIRES2-EGFP-TNFα and curcumin for pancreatic cancer treatment. Due to the internal hydrophobic cavity of amylose component, ADP could load efficiently curcumin with anticancer activity and showed a sustained release behavior. Moreover, the curcumin-loaded ADP could form colloidally stable nanocomplexes with plasmid DNA in aqueous system due to the existence of cationic poly( l -lysine) dendrons and exhibited high gene transfection efficiency. The in vitro and in vivo tests confirmed the effectiveness of using ADP to co-deliver plasmid pIRES2-EGFP-TNFα and curcumin for synergistic therapy of pancreatic cancer.
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ANALYTICAL CHEMISTRY
·
2023
·
IF 7.4
Fanglei Liu,Jiayi Zhao,Xinyu Liu,Xi Zhen,Qiumei Feng,Yingqiu Gu,Guohai Yang,Lulu Qu,Jun-Jie Zhu
In this paper, a photoelectrochemical (PEC)-surface-enhanced Raman scattering (SERS) dual-mode biosensor is constructed coupled with a dual-recognition binding-induced DNA walker with a carbon nitride nanosheet (C3N4)/MXene-gold nanoparticles (C/M-Au NPs) accelerator, which is reliable and capable for sensitive and accurate detection of Staphylococcus aureus (S. aureus). Initially, a photoactive heterostructure is formed by combining C3N4 and MXene via a simple electrostatic self-assembly as they possess well-matched band-edge energy levels. Subsequently, in situ growth of gold nanoparticles on the formed surface results in better PEC performance and SERS activity, because of the synergistic effects of surface plasmon resonance and Schottky barrier. Furthermore, a three-dimensional, bipedal, and dual-recognition binding-induced DNA walker is introduced with the formation of Pb2+-dependent DNAzyme. In the presence of S. aureus, a significant quantity of intermediate DNA (I-DNA) is generated, which can open the hairpin structure of Methylene Blue-tagged hairpin DNA (H-MB) on the electrode surface, thereby enabling the switch of signals for the quantitative determination of S. aureus. The constructed PEC-SERS dual-mode biosensor that can be mutually verified under one reaction effectively addresses the problem of the low detection accuracy of traditional sensors. Experimental results revealed that the effective combination of PEC and SERS is achieved for amplification detection of S. aureus with a detection range of 5–108 CFU/mL (PEC) and 10–108 CFU/mL (SERS), and a detection of limit of 0.70 CFU/mL (PEC) and 1.35 CFU/mL (SERS), respectively. Therefore, this study offers a novel and effective dual-mode sensing strategy, which has important implications for bioanalysis and health monitoring.
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ACS Nano
·
2023
·
IF 17.1
Shaoguang Li,Jun Dai,Man Zhu,Netzahualcóyotl Arroyo-Currás,Hongxing Li,Yuanyuan Wang,Quan Wang,Xiaoding Lou,Tod E. Kippin,Shixuan Wang,Kevin W. Plaxco,Hui Li,Fan Xia
The ability to track the levels of specific molecules, such as drugs, metabolites, and biomarkers, in the living body, in real time and for long durations, would improve our understanding of health and our ability to diagnose, treat, and monitor disease. To this end, we are developing electrochemical aptamer-based (EAB) biosensors, a general platform supporting high-frequency, real-time molecular measurements in the living body. Here we report that the use of an agarose hydrogel protective layer for EAB sensors significantly improves their signaling stability when deployed in the complex, highly time-varying environments found in vivo. The improved stability is sufficient that these hydrogel-protected sensors achieved good baseline stability and precision when deployed in situ in the veins, muscles, bladder, or tumors of living rats without the use of the drift correction approaches traditionally required in such placements. Finally, our implantable gel-protective EAB sensors achieved good biocompatibility when deployed in vivo in the living rats without causing any severe inflammation.
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Experimental and Therapeutic Medicine
·
2021
·Vol. 22
·期 1
·1-8
·
IF 2.447
Peng Yi,Xiongfeng Xu,Jiawei Yao,Bo Qiu
Genetics and epigenetics are important subjects in the field of osteoarthritis (OA) research. DNA methylation may affect gene transcription, but the specific mechanisms have remained to be fully elucidated. In the present study, the ChAMP methylation analysis package was used to identify differentially methylated genes (DMGs) from the dataset GSE63695 from the Gene Expression Omnibus (GEO) database. The distribution of differentially methylated sites (DMS) and the total array sites across the genome were analyzed by enrichment analysis. Subsequently, two mRNA expression profiling datasets, GSE114007 and GSE113825, were obtained from the GEO database and common differentially expressed genes (DEGs) were identified using the Limma package. Key genes were screened by analyzing the distribution of DMS across the genome consisting of DEGs and DMGs. A total of 1,662 and 1,986 DEGs were identified between OA and normal human cartilage from the GSE113825 and GSE114007 dataset, respectively. A further screening revealed 292 genes with common differences between the two datasets. A total of 574 DMS containing 394 DMGs were observed between OA and normal cartilage. Integrative analysis revealed a corresponding subset of 15 genes. Of these, 6 genes were verified by reverse transcription‑quantitative PCR, confirming that the mRNA expression of 5 genes (MAP1B, FNDC1, ANLN, SCNN1A and STC2) in OA cartilage was consistent with the mRNA expression from the analysis of the datasets. Upon treatment with the DNA methylation inhibitor 5‑aza‑2'‑deoxycytidine, the mRNA levels of FNDC1 and SCNN1A were decreased, and no significant alteration in the mRNA levels of MAP1B, ANLN, KCNN4 and STC2 was observed. The incidence of differential methylation varied in subregions of the genome and the effects on transcription were associated with the distribution of DEGs across the genome. The regulation of this appears more complex than initially postulated. Combining the data on epigenetic differences of OA with the genome or transcriptome data for analysis may improve the understanding of the pathophysiological processes of OA. FNDC1 and SCNN1A may potentially be valuable biomarkers for OA.
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SENSORS AND ACTUATORS B-CHEMICAL
·
2021
·
IF 7.46
Jiao Zhou,Min Qing,Yu Ling,Lei Wang,Nian Bing Li,Hong Qun Luo
DNAzyme and G-quadruplex are purposed as universal recognition elements to achieve multi-functional signal amplification and readout, showing great potential in the design of biosensors and nanodevices. Herein, the single-stranded DNA (ssDNA) opens the dumbbell-shaped hairpin probes, the generated double-stranded DNA (dsDNA) is constructed as a nanobridge, and the released G-rich sequence is folded into a G-quadruplex village. The fluorescence of crystal violet/G-quadruplex complex villages is enhanced by the dsDNA nanobridge connections. Under the excitation wavelength of 580 nm, the fluorescence emission intensity at 624 nm exhibits a good linearity with the concentration of crystal violet (CV) in the range of 10–250 nM, which can be used for sensitive detection of CV . Additionally, combined with Pb 2+ -dependent DNAzyme, a platform for quantitative detection of Pb 2+ is established. In the concentration range of 5 nM – 25 μM, the fluorescence signal increases with the increase of Pb 2+ concentration. The quantitative detection limit of Pb 2+ by this method is 5 nM, and it possesses good selectivity and recovery for Pb 2+ in a variety of water samples. More importantly, the platform can sensitively detect various metal ions by changing the DNAzyme sequence. It has potential application in the field of environmental monitoring.
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ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2021
·Vol. 413
·期 8
·2071-2079
·
IF 4.157
Zhao Yunwei,Zhang Hui,Wang Ying,Zhao Yanfang,Li Yaowei,Han Lei,Lu Lihua
A simple fluorescence detection platform has been established for acetamiprid assay based on DNA three-way junctions (TWJs), which can triple the fluorescence signal without any other amplification. It is designed with three single-stranded DNAs (ssDNA), each of which contains one-third or two-thirds of the G-quadruplex sequence at each end. Upon the addition of acetamiprid, the conformation of the aptamer-containing double-stranded DNA (dsDNA) changes from its original conformation and releases a strand of ssDNA. This ssDNA, with the other two ssDNAs, can assemble into DNA TWJs, and the three pairs of the branched ends of the DNA TWJs are adjacent to each other, allowing them to form three units of G-quadruplexes. Hence, the fluorescence of N -methyl mesoporphyrin IX (NMM) is lighted by the nascent G-quadruplexes. Graphene oxide (GO) is then added to minimize the detection background by absorbing the free NMM and non-target-induced ssDNA. The proposed strategy can assay acetamiprid in a wide linear range of 0–500 nM with a detection limit of 5.73 nM. More importantly, this assay platform demonstrates high potential for acetamiprid assay in food control and environmental monitoring.
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JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2021
·
IF 5.279
Huan Li,Jiesi Feng,Yanbo Wang,Guangming Liu,Xiaojing Chen,Linglin Fu
DNA-based detection methods are highly promising for risk assessment in the food sector, such as tracing the existence of food allergens. However, due to the complexity of food matrices, cumbersome protocols are often needed to isolate the DNA components, which hinder the achievement of rapid and on-site detection. Herein, an instant and multiple DNA extraction method was developed based on the poly(vinyl alcohol) microneedle (MN) patch. With simple press and peel-off operations within 1 min, samples suitable for DNA-based analysis such as polymerase chain reaction (PCR) could be collected. By further combining with the recombinase polymerase amplification assay, rapid screening of the allergenic risks in complex samples such as shrimp ball and cheesecake could be achieved within 30 min. The MN-based DNA extraction method not only was a potential alternative to the traditional DNA extraction method but provided a transformative approach in realizing rapid, on-site detection of foodborne hazards in collaborating with fast DNA-based assays.
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