ELECTROCHIMICA ACTA
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2015
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IF 4.504
Tingting Zhang,Huimin Zhao,Xie Quan,Shuo Chen
An electrochemiluminescence (ECL) sensing for sensitive assaying of active DNA glycosylase was developed using the enhanced host-guest recognition technique. The α-cyclodextrin was selected as the host molecule which captured the guest-labeled ECL probe to the surface of electrode by the host-guest recognition. The ECL probe can be protected from the digestion of exonuclease I (Exo I) and exonuclease III (Exo III) with the presence of the target enzyme human 8-oxoguanine DNA glycosylase 1 (hOGG 1), resulting in the ECL emission intensity was correlated with the quantity of hOGG 1. The α-cyclodextrin functionalized gold/silica (Au/SiO 2 ) cell-shell nanoparticles was prepared to enhance the host-guest recognition sensitivity. Because of the increased recognition sites provided by α-cyclodextrin functionalized Au/SiO 2 cell-shell nanoparticles, the nanoparticle-modified electrode displayed a high capacity for the guest ECL probe, and four fold enhancement of the ECL signals was achieved. The as-prepared ECL sensing exhibits excellent analytical property for the detection of hOGG 1 in the linear range of 2–100 U mL −1 with a detection limit of 0.225 U mL −1 (S/N = 3). Interference tests show that the ECL intensity of the interferents human apurinic/apyrimidinic endonuclease (APE 1), T4 endonuclease V (T4 PDG) and endonuclease III (Endo III) is within 36.5 to 44.7% of that of hOGG 1. The ECL sensing exhibits long-term stability with a relative standard deviation (RSD) of 1.6% for 16 cycles of continuous potential scans, and the life time of the ECL sensing is up to 20 days. The obtained results in this study indicate that the proposed ECL sensing possesses high sensitivity, specificity and stability and provides a powerful tool for assaying hOGG 1 activity.
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高影响因子
JOURNAL OF COLLOID AND INTERFACE SCIENCE
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2015
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IF 3.368
Linlin Wang,Shaopu Liu,Wanjun Liang,Dan Li,Jidong Yang,Youqiu He
A fluorescent “turn off–on” pattern for the detection of herring sperm DNA (hsDNA) had been designed through utilizing the interaction between polymyxin B sulfate (PMBS) and hsDNA as an inherent performance and the fluorescent transformation of glutathione (GSH)-capped CdTe quantum dots (QDs) as an external manifestation. Due to the occurrence of the photoinduced electron transfer from the QDs to PMBS, the fluorescence of GSH-capped CdTe QDs could be effectively quenched by PMBS, causing the system into “off” state. With the addition of hsDNA, the quenched fluorescence of GSH-capped CdTe QDs could be restored for the reason that PMBS embedded into hsDNA double helix structure to form new complex and peeled off from the surface of GSH-capped CdTe QDs, leading the system into “on” condition. Corresponding experimental results illustrated that the relative recovered fluorescence intensity of GSH-capped CdTe QDs–PMBS system was near proportional to the concentration of hsDNA within the range of 0.059–15.0 μg mL −1 . This proposed method demonstrated a good linear correlation coefficient of 0.9937 and a detection limit (3 σ / K ) of 0.018 μg mL −1 for hsDNA. This dual-directional fluorescent biosensor overcame the selectivity problem commonly existed in the traditional mono-directional fluorescence detection mode and owned perfect analysis applications in biochemical DNA monitoring.
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BIOSENSORS & BIOELECTRONICS
·
2015
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IF 6.409
Ying Zhu,Xiao-chun Hu,Shuo Shi,Ru-ru Gao,Hai-liang Huang,Yan-yan Zhu,Xiao-yan Lv,Tian-ming Yao
We report here an ultrasensitive strategy based on the recognition-induced conformational alteration of aptamer and fluorescence turn-on abilities of guanine-rich (G-rich) DNA sequence in proximity to silver nanoclusters for adenosine triphosphate (ATP), adenosine (A) and thrombin (TB) detection. Herein, we designed two tailored DNA sequences noted as complementary DNA (abbreviated as c-DNA) and signal probe DNA (abbreviated as s-DNA), respectively. c-DNA is designed as a special structure consisting of a sequence complementary to aptamer at the 3′-end and a guanine-rich DNA sequence at the 5′-end; s-DNA contains a cytosine-rich sequence responsible for Ag NCs templated synthesis at the 3′-end and a link sequence (part of aptamer) complementary to partial of the c-DNA at the 5′-end. In the presence of target, the aptamer associated with the target, resulting in the formation of duplex DNA (dsDNA), the DNA–Ag NCs thereafter could close to the guanine-rich sequence, leading to enhanced fluorescence signal readout. The widespread application of the sensing system is achieved success in the detection of three biomolecules . ATP, adenosine and thrombin in the range of 0.5-8.0 μM, 0.5-7.0 μM and 50-900 nM could be linearly detected with the detection limits of 91.6 nM, 103.4 nM and 8.4 nM, respectively. This label-free and turn-on fluorescent sensing system employing the mechanism proposed here turns out to be sensitive, selective, and convenient for the detection of biomolecules without washing and separation steps.
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高影响因子
ACS Applied Materials & Interfaces
·
2015
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IF 6.723
Xuehong Min,Yuan Zhuang,Zhenyu Zhang,Yongmei Jia,Abdul Hakeem,Fuxin Zheng,Yong Cheng,Ben Zhong Tang,Xiaoding Lou,Fan Xia
We demonstrate an ultrasensitive microRNA detection method based on an extremely simple probe with only fluorogens but without quencher groups. It avoids complex and difficult steps to accurately design the relative distance between the fluorogens and quencher groups in the probes. Furthermore, the assay could accomplish various detection limits by tuning the reaction temperature due to the different activity of exonuclease III corresponding to the diverse temperature. Specifically, 1 pM miR-21 can be detected in 40 min at 37 °C, and 10 aM (about 300 molecules in 50 μL) miR-21 could be discriminated in 7 days at 4 °C. The great specificity of the assay guarantees that the real 21 urine samples from the bladder cancer patients are successfully detected by our method.
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BIOSENSORS & BIOELECTRONICS
·
2013
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IF 5.437
Huanshun Yin,Bing Sun,Yunlei Zhou,Mo Wang,Zhenning Xu,Zhengliang Fu,Shiyun Ai
In this work, we fabricated a novel photoelectrochemical immunosensor for assay of DNA methylation , where Bi 2 S 3 nanorods were used as photoelectric conversion material, MBD1 protein (a kind of methyl bonding domain protein) was used as DNA methylation recognizing unit, anti-his tag antibody was used to further inhibit the photocurrent and increase the detection sensitivity. The results demonstrated that Bi 2 S 3 possessed excellent photoelectron property. The detection conditions, such as Bi 2 S 3 concentration, MBD1 protein concentration, incubation time of MBD1 protein, antibody concentration and antibody incubation time, were optimized. Under optimal experimental conditions, the photocurrent variation was proportional to the logarithm of methylated target DNA concentration from 10 −9 to 10 –13 M with detection limit of 3.5×10 –14 M ( S / N =3). Moreover, the immunosensor presented high detection specificity, even distinguishing single-base mismatched sequence.
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JOURNAL OF CONTROLLED RELEASE
·
2016
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IF 7.441
Wenjun Zhang,Feihu Wang,Yun Wang,Jining Wang,Yanna Yu,Shengrong Guo,Rongjun Chen,Dejian Zhou
A thiolated pH-responsive DNA conjugated gold nanorod (GNR) was developed as a multifunctional nanocarrier for targeted, pH-and near infrared (NIR) radiation dual-stimuli triggered drug delivery. It was further passivated by a thiolated poly(ethylene glycol)-biotin to improve its cancer targeting ability by specific binding to cancer cell over-expressed biotin receptors. Doxorubicin (DOX), a widely used clinical anticancer drug , was conveniently loaded into nanocarrier by intercalating inside the double-stranded pH-responsive DNAs on the GNR surface to complete the construction of the multifunctional nanomedicine . The nanomedicine can rapidly and effectively release its DOX payload triggered by an acidic pH environment (pH ~ 5) and/or applying an 808 nm NIR laser radiation. Compared to free DOX, the biotin-modified nanomedicine displayed greatly increased cell uptake and significantly reduced drug efflux by model multidrug resistant (MDR) breast cancer cell lines (MCF-7/ADR). The application of NIR radiation further increased the DOX release and facilitated its nuclear accumulation. As a result, this new DNA-GNR based multifunctional nanomedicine exerted greatly increased potency (~ 67 fold) against the MDR cancer cells over free DOX.
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SENSORS AND ACTUATORS B-CHEMICAL
·
2016
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IF 4.758
Qian Luan,Yun Xi,Ning Gan,Yuting Cao,Tianhua Li,Yinji Chen
Herein, a promising colorimetric strategy for small molecule detection was developed based on one novel magnetic single-stranded DNA binding protein (SSB)-linked composite probe. Firstly, a magnetic probe was synthesized by labeling SSB on gold magnetic particles (Au MNPs-SSB) and detection probe by co-immobilizing aptamer (Apt) and horseradish peroxidase (HRP) on the core-shell SiO 2 @Au nanoparticles (Apt-SiO 2 @Au-HRP); furthermore, Au MNPs-SSB was bound with Apt-SiO 2 @Au-HRP to construct a composite probe relied on the specific affinity between SSB and free aptamer. When the target, with chloramphenicol (CAP) as model, was introduced into the reaction system, the formation of the target-Apt-SiO 2 @Au-HRP complex can release from its parent probe to supernatant after magnetic separation because SSB can’t bind target-aptamer complex. The target-Apt-SiO 2 @Au-HRP complex in supernatant could efficiently catalyze 3,3′,5,5′-tetramethylbenzidine(TMB)-H 2 O 2 system for the color development. Under the optimized reaction conditions, the linear range was from 0.05 to100 ng mL −1 with a low detection limit of 0.02 ng mL −1 for CAP. Moreover, the aptamer assay was employed to detect CAP in milk and the results were consistence with those from ELISA. In addition, the assay can be easily extended to other small molecules detection when changing the corresponding aptamer on the detection probe.
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ANALYTICAL CHEMISTRY
·
2016
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IF 5.886
Hui Zhang,Huilei Dong,Guoqing Yang,Hongfei Chen,Chenxin Cai
Effective detection of DNA methyltransferase (DNMT) activity is significant for cancer research. Herein, we developed a sensitive electroanalytical method to detect human DNA (cytosine-5)–methyltransferase 1 (DNMT1) from crude lysates of cancer cells. In this assay, capture DNA having a preferred DNMT1 methylation site was immobilized on a gold electrode and then hybridized with gold nanoparticle (Au NP)–DNA complexes. The modified electrodes were equilibrated with the lysate and then incubated with methylation-sensitive restriction enzyme. If the lysate was negative for DNMT1 activity, the Au NP–DNA complexes would be cut by the restriction enzyme and released from the electrode. Conversely, restriction enzyme cleavage would be blocked by the fully methylated duplexes, and the Au NP–DNA complexes would remain on the electrode. Electroactive Ru(NH3)63+ was used as the signal reporter, because of its electrostatic attraction to DNA, resulting in an electrochemical signal. Since the electrochemical signal reflects the amount of Ru(III) redox and the amount of Ru(III) redox is correlated with the activity of DNMT1, the activity of DNMT1 is proportional to the electrochemical signal. The signal could be amplified by the numerous DNAs on the Au NPs and further amplified by Ru(III) redox recycling. With this method, a detection limit down to 0.3 U/mL for pure DNMT1 and 8 MCF-7 cells was achieved. DNMT1 activities of different cell lines were also successfully evaluated.
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ANALYTICAL CHEMISTRY
·
2024
·
IF 6.7
Junhua Chen,Xu Wang,Junhui Guo,Yiwen Lv,Manjia Chen,Hui Tong,Chengshuai Liu
The uranyl ion (UO22+) is the most stable form of uranium, which exhibits high toxicity and bioavailability posing a severe risk to human health. The construction of ultrasensitive, reliable, and robust sensing techniques for UO22+ detection in water and soil samples remains a challenge. Herein, a DNA network biosensor was fabricated for UO22+ detection using DNAzyme as the heavy metal recognition element and double-loop hairpin probes as DNA assembly materials. UO22+-activated specific cleavage of the DNAzyme will liberate the triggered DNA fragment, which can be utilized to launch a double-loop hairpin probe assembly among Hab, Hbc, and Hca. Through multiple cyclic cross-hybridization reactions, hexagonal DNA duplex nanostructures (n[Hab•Hbc•Hca]) were formed. This DNA network sensing system generates a high fluorescence response for UO22+ monitoring. The biosensor is ultrasensitive, with a detection limit of 2 pM. This sensing system also displays an excellent selectivity and robustness, enabling the DNA network biosensor to work even in complex water and soil samples with excellent accuracy and reliability. With the advantages of enzyme-free operation, outstanding specificity, and high sensitivity, our proposed DNA network biosensor provides a reliable, simple, and robust method for trace levels of UO22+ detection in environmental samples.
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SENSORS AND ACTUATORS B-CHEMICAL
·
2017
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IF 5.401
Yilin Lu,Jie Zhong,Guohua Yao,Qing Huang
A prototype of label-free SERS sensor, based on the formation of T-Hg 2+ -T pairs and the corresponding orientation variation of DNA on Au shell surface, was developed for detection of Hg 2+ in aqueous solution quantitatively and selectively. The DNA aptamer immobilized on the surface of SiO 2 @Au core/shell nanoparticles (NPs) consisted of two segments, namely, the segment which has the consecutive thymines (T) severing as the Hg 2+ recognition elements, and the segment which contains the guanine (G) and adenine (A) bases working as the signal reporter. With interaction of Hg 2+ ions and the thymines between the adjacent single-stranded DNA (ssDNA), the DNA molecule adopted vertical orientation, resulting in increase of Raman intensity ratio I(660 cm −1 )/I(736 cm −1 ) with increase of Hg 2+ concentration, which thus allowed to measure trace amounts of Hg 2+ in aqueous solution selectively and quantitatively. Our results revealed that this label-free SERS sensor could sensitively respond to Hg 2+ ions within a wide concentration range (from 1 × 10 −8 to 1 × 10 −3 M). This work therefore demonstrates that proper design of aptamer-modified SiO 2 @Au core/shell NPs can be utilized for label-free SERS detection of heavy metal ions in the environment.
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MOLECULAR MEDICINE
·
2025
·Vol. 31
·期 1
·1-15
·
IF 6.4
Pei Lipeng,Zhu Yan,Li Wenpeng,Xu Bin,Sun Jingli
Background Uterine corpus endometrial carcinoma (UCEC) is one of the most frequent female genital malignant tumors. Targeting DNA damage and cell apoptosis are regarded as effective ways for UCEC therapy. Celastrol is a natural anti-cancer product from the Celastraceae plant family, while its role in UCEC has not been investigated. Methods UCEC cell lines Ishikawa and HEC-1-A were applied and treated with different concentrations of Celastrol. The appropriate and nontoxic concentrations were used for the subsequent experiments. Functional experiments analyzed the cell viability, cell cycle distribution, DNA damage, apoptosis and the expression of related proteins. We determined tumor growth in xenograft nude mice. Bioinformatic analysis, protein coimmunoprecipitation (Co-IP), luciferase assay, cell experiments were performed to reveal the relationship of Celastrol/KAT2B/RBPJ/MCM4 in UCEC. Results Treatment of Celastrol inhibited cell viability in a dose-dependent manner, and caused cell cycle arrest, accompanied by the downregulation of CDK2 and cyclin E expression and the upregulation of p21. Celastrol treatment resulted DNA damage and apoptosis in cultured cells, as demonstrated by increased number of TUNEL-positive cells, activity of caspase-3 and expression of cleaved-caspase-9, cleaved PARP1 and γ-H2AX. In xenograft nude mice, Celastrol also repressed tumor growth. Furthermore, lysine acetyltransferase KAT2B was a putative target of Celastrol, and its expression was upregulated by Celastrol in vitro and in vivo. Overexpression of KAT2B in UCEC inhibited cell proliferation and increased DNA damage and apoptosis. KAT2B knockdown overcame the anti-proliferative and pro-apoptotic roles of Celastrol. Moreover, Co-IP demonstrated that KAT2B bound to RBPJ, a transcriptional repressor, and increased the acetylation of RBPJ. RBPJ could bind to the MCM4 promoter to suppress the luciferase activity. Further functional analysis revealed that the functions of KAT2B in UCEC cell proliferation, DNA damage and apoptosis were mediated by MCM4, and Celastrol enhanced RBPJ acetylation and reduced MCM4 expression. Conclusions These results underscore that Celastrol is a promising anti-cancer agent in UCEC with preferential anti-proliferative, pro-apoptotic and DNA damage effects through the KAT2B/RBPJ/MCM4 axis, and KAT2B is a promising therapeutic target for UCEC. Graphical abstract
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BIOSENSORS & BIOELECTRONICS
·
2012
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IF 5.602
Jing Wang,Wei-Wei Zhao,Hong Zhou,Jing-Juan Xu,Hong-Yuan Chen
In this work, we firstly enunciated the presence of synergy effect between the electron and energy transfer in an electrochemiluminescence (ECL) system involving the CdS NCs and Au NPs, based on which an amplified ECL biosensor was constructed for sensitive detection of DNA–protein interactions. Specifically, Au NPs modified on electrode, following by assembled probe DNA . Then the modified GCE was hybridized with SiO 2 @CdS/DNA conjugates. With DNA duplex as a rigid spacer, Au NPs could not only accelerate the electron transfer but also produce the surface plasma resonance upon excited ECL of CdS NCs. However, the enhanced ECL could be greatly suppressed by the binding of target protein. The prepared biosensor possesses excellent analytical performance with the linear range of target protein from 0.015 to 150 nM.
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