ANALYTICAL CHEMISTRY
·
2023
·
IF 7.4
Peiqing Luo,Xiaocui Huang,Fang Luo,Zhonghui Chen,Yu Chen,Cuiying Lin,Jian Wang,Bin Qiu,Zhenyu Lin
In this work, combined with the high amplification efficiency of hybridization chain reaction (HCR), high specificity of the CRISPR/Cas12a system, and convenience of the homogeneous electrochemiluminescence (ECL) assay based on the regulation of negative charge on the reporting probes, a sensitive ECL biosensor for hepatitis B virus DNA (chosen as a model target) had been developed. The initiator chain trigger DNA that can induce HCR amplification is modified on the surface of ruthenium bipyridine-doped silica nanoparticles (Ru@SiO2 NPs) first, and large amounts of negative charges modified on the particles were achieved through the HCR amplification reaction. The efficiency of the nanoparticles reaching the negatively charged working electrode can be regulated and realize the change of the ECL signal. In addition, long DNA on the surface of the luminescent body may prevent the coreactant from entering the pore to react with ruthenium bipyridine. These factors combine to produce a low-background system. The presence of the target can activate the CRISPR/Cas12a system and make trigger DNA disappear from the nanoparticle surface, and strong ECL can be detected. The sensor does not require a complex electrode modification; therefore, it has better reproducibility. Additionally, due to dual signal amplification, the sensor has a high sensitivity. In the range of 10 fM to 10 nM, the ECL intensity exhibits a strong linear relationship with the logarithm of the target concentration, and the detection limit is 7.41 fM. This sensor has shown high accuracy in detecting clinical samples, which holds significant potential for application in clinical testing.
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MICROCHIMICA ACTA
·
2019
·Vol. 187
·期 1
·1-8
·
IF 5.479
Liu Yue,Liu Wei-Jun,Hu Jia,Li Yan,Wang Ying,Zhao Li-Xia
The target-triggered DNA assembling probe is presented for highly selective protein detection. Target-triggered DNA assembling is used in an amplification strategy based on affinity binding for identification and determination of proteins in general. Specifically, it was applied to the platelet derived growth factor-BB (PDGF-BB). A hairpin DNA (H-DNA) probe was designed containing (a) an aptamer domain for protein recognition and (b) a blocked DNAzyme domain for DNAzyme cleavage. An assistant DNA (A-DNA) probe containing aptamer and complementary domains was also employed to recognize protein and to induce DNA assembly. Once H-DNA and A-DNA recognize the same protein, H-DNA and A-DNA are in close proximity to each other. This induces DNA assembling for protein-triggered complex (Protein-Complex) with free DNAzyme domains. The free DNAzymes trigger the circular cleavage of molecular beacons for amplified signals. The assay is performed by fluorometry at an excitation wavelength of 980 nm and by collecting fluorescence at 545 nm. The platelet derived growth factor-BB (PDGF-BB) was accurately identified and selectively determined by this assay with a 22 pM detection limit (using the 3 σ criterion). The responses for PDGF-BB is nearly 6-fold higher than for PDGF-AB, and 16-fold higher than PDGF-AA. This upconversion assay avoids any interference by the autofluorescence of biological fluids.
Schematic representation of the principle of the target-triggered DNA assembling probes mediated amplification strategy based on affinity binding for PDGF-BB. The UCNP probe is used for the quantitation of PDGF-BB with high selectivity.
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INDUSTRIAL & ENGINEERING CHEMISTRY RESEARCH
·
2015
·
IF 2.587
Xiaoqian Chen,Qingxiang Wang,Liheng Wang,Hongxu Guo,Yizhen Yang,Feiming Li,Feng Gao
Rugby-ball-shaped CoS2 (rCoS2) submicroparticles have been synthesized via a simple hydrothermal reaction. The rCoS2 submicroparticles were then casted on a glassy carbon electrode and served as a highly conductive and large surface area platform for the electrosynthesis of poly(2-thiophenesulfonyl chloride) (pTSC), using 2-thiophenesulfonyl chloride as the monomer. The stepwise modification process was characterized by attenuated total reflectance–Fourier transform infrared spectroscopy (ATR-FTIR) and different electrochemical technologies. The stable pTSC layer was utilized as a functional interface to immobilize amino-modified probe DNA via a facile sulfonyl–amino condensation reaction, without the assistance of any other cross-linkers. Because of the synergistic effect of rCoS2 and pTSC from the hybrid material, the developed DNA biosensor could detect target DNA over a wide dynamic range, from 1.0 × 10–13 to 1.0 × 10–7 M, with the detection limit down to 1.1 × 10–14 M. Benefiting from the steady electropolymerization film of pTSC and the stable sulfamide bond between the amino-modified DNA and pTSC, the biosensor also showed high stability, reusability, and reproducibility.
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JOURNAL OF CHROMATOGRAPHY A
·
2024
·
IF 3.8
Ying Shi,Yi-Wen Wu,Qian-Xue Shen,Jun Cao
Aristolochic acids are one of the major compounds in aristolochia plants, which are nephrotoxic and carcinogenic. A method was established for the detection and identification of aristolochic acids and their DNA adducts in four different herbs using ultra-high performance liquid chromatography-ion mobility quadrupole time-of flight mass spectrometry. Solid phase extraction conditions were optimized to improve the sensitivity of the experiment by using 40 mg of C 18 as adsorbent and 100 μL ethanol as elution solvent. At a collision energy of 10-40 eV, these compounds and cleavage patterns were precisely identified and analyzed by secondary fragmentation and collision cross section values. The obtained mass spectrometry data were then analyzed by targeted metabolomics, including principal component analysis, partial least squares-discriminant analysis and hierarchical clustering analysis, and importing the samples in the established model, the confidence values can reach 0.61 and 0.76. All in all, this method can provide a useful tool for the detection of aristolochic acids and deoxyribonucleic acid adducts. In conclusion, this method was successfully used for the detection and identification of aristolochic acids and their DNA adducts.
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高影响因子
MICROCHIMICA ACTA
·
2019
·Vol. 186
·期 3
·1-8
·
IF 5.479
Liu Shi Gang,Luo Dan,Han Lei,Li Nian Bing,Luo Hong Qun
A hybrid material composed of guanine-rich single stranded DNA (G-rich ssDNA) and cobalt oxyhydroxide (CoOOH) nanosheets is used as a nanoprobe for fluorometric turn-on detection of ascorbic acid (AA). The CoOOH nanosheets function as a recognition component for AA. The G-rich ssDNA is used to produce a G-quadruplex, and the G-quadruplex/thioflavin T (ThT) complex acts as a fluorescent reporter. In the absence of AA, p-phenylenediamine (PPD) is oxidized to form oxPPD which has a dark red color. It causes the fluorescence of the G-quadruplex/ThT complex to be quenched. However, in the presence of AA, the CoOOH nanosheets of the nanoprobe are preferentially reduced by AA. Hence, PPD is not oxidized, and fluorescence is not quenched. A fluorometric turn-on method was developed based on these findings. It has a detection limit of 94 nM and works in the concentration range from 1 to 10 and 20 to 80 μM. This method was applied to the determination of AA in (spiked) fruit juice samples.
Schematic presentation of a fluorescent assay of ascorbic acid (AA) is established using a nanoprobe composed of guanine-rich single stranded DNA (G-rich ssDNA) and cobalt oxyhydroxide (CoOOH) nanosheets. It is based on competitive reduction of CoOOH by p-phenylenediamine (PPD) and AA. Thioflavine T (ThT) induces the formation of fluorescent G-quadruplex/ThT complex. The oxidized form of PPD (oxPPD) can quench the fluorescence via fluorescence resonance energy transfer (FRET), but AA suppresses quenching.
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高影响因子
CURRENT CANCER DRUG TARGETS
·
2022
·Vol. 22
·期 9
·771-783
·
IF 2.907
Cui Zhi,Sun Shiqun,Li Jia,Li Jianing,Sha Tong,He Jie,Zuo Linjing
Background: Inhibitor of growth 4 (ING4) level was reported to be decreased in head and neck squamous cell carcinoma (HNSC) tissue, however, it is unknown whether and how ING4 participates in regulating the development of oral squamous cell carcinoma (OSCC). Objective: This study
aimed to investigate the role and mechanism of ING4 in OSCC. Methods: ING4 was forced to up- or down-regulated in two OSCC cell lines, and its effects on the malignant behavior of OSCC cells were investigated in vitro. The ubiquitination level of NF-kB p65 in ING4 upregulated cells was
measured by co-immunoprecipitation. Moreover, the effects of ING4 on the methylation level of ALDH1A2 were evaluated by methylation-specific polymerase chain reaction (MSP) assay. The role of ING4 in OSCC growth in vivo was observed in nude mice. Results: Our results showed that the expression
of ING4 in OSCC cell lines was lower than that in normal oral keratinocyte cells. In vitro, ING4 overexpression inhibited the proliferation, migration, and invasion of OSCC cell lines and ING4 silencing exhibited opposite results. We also demonstrated that ING4 overexpression promoted the
ubiquitination and degradation of P65 and reduced DNA methyltransferase 1 (DNMT1) expression and Aldehyde dehydrogenase 1A2 (ALDH1A2) methylation. Moreover, overexpression of p65 rescued the suppression of malignant behavior, induced by ING4 overexpression. In addition, ING4 negatively regulated
the growth of OSCC xenograft tumors in vivo. Conclusion: Our data evidenced that ING4 played a tumor-repressing role in OSCC in vivo and in vitro via NF-ΚB/DNMT1/ALDH1A2 axis.
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JOURNAL OF NANOBIOTECHNOLOGY
·
2018
·Vol. 16
·期 1
·1-10
·
IF 5.294
Wang Shuai,Qu Fei,Han Wenli,You Jinmao
Background Differentiation of telomere length is of vital importance because telomere length is closely related with several deadly diseases such as cancer. Additionally, G-quadruplex and i-motif formation in telomeric DNA have been shown to act as a negative regulator of telomere elongation by telomerase in vivo and are considered as an attractive drug target for cancer chemotherapy. Results In this assay, Ag nanoclusters templated by hyperbranched polyethyleneimine (PEI–Ag NCs) are designed as a new novel resonance Rayleigh scattering (RRS) probe for sensitive differentiation of telomere length and monitoring special motifs (G-quadruplex and i-motif). In this assay, free PEI–Ag NC probe or DNA sequence alone emits low intensities of RRS, while the formation of PEI–Ag NCs/DNA complexes yields greatly enhanced RRS signals; however, when PEI–Ag NCs react with G-quadruplex or i-motif, the intensities of RRS exhibit slight changes. At the same concentration, the enhancement of RRS signal is directly proportional to the length of telomere, and the sensitivity of 64 bases is the highest with the linear range of 0.3–50 nM (limit of detection 0.12 nM). On the other hand, due to the conversion of telomere DNA molecules among multiple surrounding conditions, a DNA logic gate is developed on the basis of two chemical input signals (K + and H + ) and a change in RRS intensity as the output signal. Conclusion Our results indicate that PEI–Ag NCs can serve as a novel RRS probe to identify DNA length and monitor G-quadruplex/i-motif through the different increasing degrees of RRS intensity. Meanwhile, the novel attributes of the nanoprobe stand superior to those involving dyes or labeled DNA because of no chemical modification, low cost, green, and high efficiency.
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