ELECTROPHORESIS
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2020
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IF 3.081
Mengqi Li,Hao Shen,Zixin Zhou,Wenting He,Ping Su,Jiayi Song,Yi Yang
In recent years, CE-integrated immobilized enzyme reactors (IMERs) for single-enzyme immobilization have attracted considerable attention. However, there has been little research on multienzyme immobilization in CE. Here, we introduce a method for fabricating a CE-integrated IMER, using DNA-directed immobilization to fix glucose oxidase and horseradish peroxidase in the capillary, which had been functionalized with polyamidoamine dendrimer (PAMAM). Owing to the reversibility of DNA hybridization, the reactor is capable of dynamic immobilization. Moreover, by introducing the PAMAM, the loading capacity of the IMER is greatly enhanced, and the PAMAM can spontaneously form complexes with DNA and then contribute to the efficiency and stability of the reactor. After 25 days storage, the prepared IMER ultimately retained approximately 70% of its initial activity. We also used the IMER to detect glucose, and the favorable linearity was obtained over the concentration range of 0.78–12.5 mM, with an LOD of 0.39 mM, demonstrating that the CE-integrated IMER can be applied to actual samples. We believe that this strategy can be extended to other multienzyme immobilization systems, and CE-integrated IMERs are potentially useful in a wide range of biochemical research applications.
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BIOSENSORS & BIOELECTRONICS
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2020
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IF 10.257
Yanyan Yu,Wei S. Zhang,Yuehua Guo,Huaping Peng,Min Zhu,Dandan Miao,Gaoxing Su
Tumor-derived exosomes containing multiple proteins originating from parent cancer cells have emerged as biomarkers for cancer diagnosis. Herein, we propose a three-dimensional DNA motor-based exosome assay platform for the selective and sensitive detection of exosomes. The DNA motor used gold nanoparticle (GNP) tracks, consisting of fluorescein-labeled substrate strands and aptamer-locked motor strands. Recognition of the target protein on exosomes by its aptamer unlocked the motor strand and triggered the DNA motor process. Powered by restriction endonuclease , the motor strands autonomously walked along the GNP track. Each movement step cleaved one substrate strand and restored one fluorescein molecule. For exosome detection, the proposed method displayed a broad dynamic range acrossing 5 orders of magnitude with the detection limit as low as 8.2 particles/μL in PBS. The method also exhibited good selectivity among different tumor-derived exosomes and performed well in complex biological samples. The capability to profile exosomal surface proteins efficiently endowed our DNA motor great potential for developing a simple and cost-effective device for clinical diagnosis.
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BIOMEDICINE & PHARMACOTHERAPY
·
2019
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IF 3.743
Tianhua Liu,Ling Zuo,Dongqing Guo,Xinlou Chai,Jie Xu,Zhaorui Cui,Zhiyi Wang,Chunying Hou
Lung cancer is the leading cause of cancer-related deaths. Ginsenoside Rg3 is the main ingredient of Ginseng which is used to treat non-small cell lung cancer (NSCLC). It has been found to enhance the efficiency of chemotherapy thereby reducing its side effects. Previous studies found that ginsenoside Rg3 can reduce the occurrence of NSCLC by inducing DNA damage. Yet, its anti-DNA damaging effects and mechanisms in tumor cells are still not fully understood. This study explored the effect of ginsenoside Rg3 on DNA repair and VRK1/P53BP1 signaling pathway. Ginsenoside Rg3 treatment significantly decreased the incidence and invasionin a mouse model of lung cancer induced by urethane. The results of cell survival assay and single cell gel electrophoresis showed that ginsenoside Rg3 protected lung adenocarcinoma cells from DNA damage as well as inhibited the proliferation of tumor cells. Ginsenoside Rg3 increased the mRNA and protein expression of VRK1 in NSCLC cells as measured by RT-qPCR and western blot, respectively. These findings suggests that ginsenoside Rg3 regulates VRK1 signaling. Immunofluorescence assays showed that P53BP1 and VRK1 protein level increased, and the VRK1 protein translocated between the nuclei and cytoplasm. Finally, this conclusion was confirmed by the reverse validation in VRK1-knockdown cells. Taken together, these results show that ginsenoside Rg3 upregulate VRK1 expression and P53BP1 foci formation in response to DNA damage thereby inhibiting the tumorigenesis and viability of cancer cells. These findings reveal the role of Rg3 in lung cancer and provides therapeutic targets for developing new drugs in the prevention and treatment of lung cancer.
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高影响因子
Nature Communications
·
2022
·Vol. 13
·期 1
·1-16
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IF 17.694
Li Fangyuan,Sun Heng,Ren Jiafeng,Zhang Bo,Hu Xi,Fang Chunyan,Lee Jiyoung,Gu Hongzhou,Ling Daishun
Platinum (Pt) resistance in cancer almost inevitably occurs during clinical Pt-based chemotherapy. The spontaneous nucleotide-excision repair of cancer cells is a representative process that leads to Pt resistance, which involves the local DNA bending to facilitate the recruitment of nucleotide-excision repair proteins and subsequent elimination of Pt-DNA adducts. By exploiting the structural vulnerability of this process, we herein report a nuclease-mimetic Pt nanozyme that can target cancer cell nuclei and induce concurrent DNA platination and oxidative cleavage to overcome Pt drug resistance. We show that the Pt nanozyme, unlike cisplatin and conventional Pt nanoparticles, specifically induces the nanozyme-catalyzed cleavage of the formed Pt-DNA adducts by generating in situ reactive oxygen species, which impairs the damage recognition factors-induced DNA bending prerequisite for nucleotide-excision repair. The recruitment of downstream effectors of nucleotide-excision repair to DNA lesion sites, including xeroderma pigmentosum groups A and F, is disrupted by the Pt nanozyme in cisplatin-resistant cancer cells, allowing excessive accumulation of the Pt-DNA adducts for highly efficient cancer therapy. Our study highlights the potential benefits of applying enzymatic activities to the use of the Pt nanomedicines, providing a paradigm shift in DNA damaging chemotherapy.
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SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
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2022
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IF 4.831
Yan Liu,Baibing Guan,Ziqi Xu,Yaohui Wu,Yonghong Wang,Ge Ning
Kanamycin was a group of essential antibiotics generally served in treating infections of animals which leached into the environment residual in food, causing health concerns. Thus, selective and sensitive monitoring of kanamycin was significant for food safety. In this work, split aptamers were used as templates to prepare fluorescent Cu/Ag NCs for detection of kanamycin. According to the impressive affinity of the aptamer to kanamycin, two different detection modes were designed using kanamycin aptamer as a recognition molecule, in which one was to combine split aptamer Apt-1 with Apt-2 to form an entangled DNA as a Cu/Ag NCs template, the other was to associate the normal aptamer after encirclement to form Cu/Ag NCs templates. After the addition of kanamycin, the fluorescence signals of the Cu/Ag NCs synthesized in the two modes were both enhanced, but the approach with split aptamer exhibited a superior observable sensitivity than that of the normal type. The detection range showed a well linear relationship between 80 nM and 10 μM when the emission wavelength was 560 nm, and the detection limit was 13.3 nM. In addition, when streptomycin, oxytetracycline, chloramphenicol and chlortetracycline were involved in the selective interference experiment under the same conditions, the fluorescence intensity of the system performed no significant changes. The results demonstrated that this method possessed favorable specificity and selectivity for the assay of kanamycin, proficiently achieving efficient, rapid and sensitive evaluation of kanamycin in the milk samples.
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JOURNAL OF NATURAL PRODUCTS
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2015
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IF 3.798
Xueqian He,Zhenzhen Zhang,Yinghan Chen,Qian Che,Tianjiao Zhu,Qianqun Gu,Dehai Li
A new, highly modified fatty acid amide, varitatin A (1), was isolated from the fungus Penicillium variabile HXQ-H-1 cultivated with the DNA methyltransferase inhibitor 5-azacytidine. The structure including the absolute configuration of 1 was established by analysis of NMR and MS data, together with chemical degradation and Mosher’s method based on MPA esters. Compound 1 showed cytotoxicity against HCT-116 cells with an IC50 value of 2.8 μM and also inhibited the effects of protein tyrosine kinases.
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MICROCHEMICAL JOURNAL
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2023
·
IF 4.8
Feng Xu,Xin Chen,Daofei Lv,Yong Liang
Here, we report a DNA hairpin strategy for electrochemical determination of Hg 2+ based on T-Hg 2+ -T pairs and exonuclease III (Exo III) assistance. The innovation of this work lies in the designed DNA structure and detection principles. Unlike the previously reported DNA hairpin structure, the ring portion of the hairpin DNA we designed consisting of T bases serves as a recognition probe, forming a T-Hg-T complex with Hg 2+ . The stem is composed of 5 pairs of complementary bases, which is conducive to the stability of the hairpin structure. After the formation of T-Hg-T in the ring, with the help of Exo III, Exo III can digest more DNA than the T bases designed in the stem. In this event, the shorter the length of the remaining DNA strand on the electrode, the weaker the repulsive force against [Fe(CN) 6 ] 3−/4− , and the stronger the differential pulse voltammetry (DPV) current signal, achieving “signal-on” transitions. Under the optimal conditions, the DPV current response increases with increasing Hg 2+ concentration and is linear with the logarithm of Hg 2+ concentration (1 nM–10 μM). The limit ofdetection is 0.16 nM. The electrochemical sensor was successfully applied to the determination of Hg 2+ in river water.
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SENSORS AND ACTUATORS B-CHEMICAL
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2023
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IF 8.4
Han Lu,Dan Wang,Guangfei Huang,Qilin Ma,Yuan Zhang,Mingliang Jin,Lingling Shui
Surface-enhanced Raman spectroscopy (SERS) has exhibited great potential in deoxyribonucleic acid (DNA) analysis. Rapid and label-free identification of single-base mutation in DNA is still limited by the sensitivity and reproducibility of SERS substrates. In this work, we develope plasmonic SiNW arrays decorated with Ag nanoparticles (SiNW@AgNP arrays) via hexadecyltrimethylammonium bromide (CTAB)-assisted galvanic reaction to address these challenges, which show excellent sensitivity and reproducibility. Under optimized experimental conditions, plasmonic SiNW@AgNP arrays enable detection of DNA bases of adenine (A), guanine (G), cytosine (C), and thymine (T) in solution at concentration down to pM-μM range, and multiple detection ability with mixture of DNA bases. More significantly, our study demonstrates the SiNW@AgNP arrays may serve as excellent SERS substrates in discriminating DNA single-base mutation between sickle cell anemia gene and normal hemoglobin gene in serum samples, which can be exploited in a wide range of plasmonic enhanced sensing applications for a fast, real-time, label-free and cost-effective clinical diagnosis.
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SENSORS AND ACTUATORS B-CHEMICAL
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2023
·
IF 8.4
Yunshan Zhang,Shijie Xu,Jian Chen,Lanyue Wang,Lina Bian,Jing ye,Lin Weng,Xiaoyu Zhao,Cheng-Te Lin,Shuang Li,Diming Zhang
Single nucleotide polymorphisms (SNPs) are emerging as important biomarkers for disease diagnosis, prognostics and disease pathogenesis. However, it is still challenging to make highly specific and sensitive SNPs detection to distinguish target DNA with single-base difference, so that flap endonuclease 1 (FEN 1) manipulated semi-DNA walker reaction and catalytic hairpin assembly (CHA)-FRET loop for ultrasensitive SNP detection has been proposed. Herein, FEN 1 was employed to specifically recognize and cut the 5′ flap of three-base overlapping structure formed by only hybridizing with mutant target (MT). Simultaneously, FEN 1 drove MT hybridization to autonomously walk along downstream probe (DP) and the cut sequence (CS) from semi-DNA walker reaction was collected by magnetic bead-based separation to avoid false-positive signal. Next, the cleaved CS triggered CHA and generated fluorescence resonance energy transfer (FRET) via Cy3 and Cy5. FEN 1 manipulated semi-DNA walker reaction to specifically recognize the target DNA and combined CHA-FRET loop to dually amplify signal. Thus, the SNP biosensor demonstrated superior performance for ultrasensitive detection of mutant KRAS gene with a low detection limit as low as 80 aM and was successfully applied to monitor the expression of mutant KRAS gene in human cancer cell lysates. This strategy provides an ultrasensitive way for the detection of biomolecules and reveals an effective avenue for diseases diagnosis.
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SCIENCE OF THE TOTAL ENVIRONMENT
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2023
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IF 9.8
Junhua Chen,Manjia Chen,Hui Tong,Fei Wu,Yizhang Liu,Chengshuai Liu
A fluorescence biosensor was developed for the ultrasensitive detection of the available lead in soil samples by coupling with DNAzyme and hairpin DNA cyclic assembly. The biorecognition between lead and 8-17 DNAzyme will cleave the substrate strands (DNA2) and release the trigger DNA (T), which can be used to initiate the DNA assembly reactions among the hairpins (H1, H2, and H3). The formed Y-shaped sensing scaffold (H1-H2-H3) contains active Mg 2+ -DNAyzmes at three directions. In the presence of Mg 2+ , the BHQ and FAM modified H4 will be cleaved by the Mg 2+ -DNAyzme to generate a high fluorescence signal for lead monitoring. The linear range of the fluorescence biosensor is from 1 pM to 100 nM and the detection limit is 0.2 pM. The biosensor also exhibited high selectivity and the nontarget competing heavy metals did not interfere with the detection results. Compare with the traditional method (DTPA+ICP-MS) for the available lead detection, the relative error (Re) is in the range from −8.3 % to 9.5 %. The results indicated that our constructed fluorescence biosensor is robust, accurate, and reliable, and can be applied directly to the detection of the available lead in soil samples without complex extraction steps.
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SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
·
2023
·
IF 4.4
Qian Zhao,Jiahao Wu,Zhenghong Jiang,Dai Lu,Xinhui Xie,Liye Chen,Xingbo Shi
Aflatoxin B1 (AFB1) is one of the most toxic mycotoxins, which is frequently detected in agricultural products. Herein, a novel functional DNA -linked immunosorbent assay (DLISA) with dual-modality based on hybrid chain reaction (HCR) has been successfully developed for ultrasensitive detection of AFB1. The strategy relies on AFB1 immune-bridged occurrence of HCR and the salt-induced aggregation of gold nanoparticles (AuNPs). An aptamer-initiator stand (Apt-Ini stand) is designed for the AFB1 recognition and the activation of HCR, which can recognize the matched hairpins and cause the crossing-opening of H1 and H2, producing a long double-stranded DNA polymer. The addition of SYBR Green I achieves the fluorescent signal output. Remaining less DNA hairpins were added and stuck on the surface of AuNPs, which were insufficient to protect the AuNPs, resulting in the salt-induced aggregation with the color change from red to blue. The dual-modality provides limits of detections of 1.333 × 10 −14 g/mL and 2.471 × 10 −15 g/mL, respectively. This DLISA with dual-modality provides not only a colorimetry that can meet the needs of on-the-spot preliminary inspection, but also a fluorescence assay that can acquire the precise results.
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MICROCHIMICA ACTA
·
2023
·Vol. 190
·期 11
·1-9
·
IF 5.7
Yao Yuying,Wang Yeru,Zhu Jingru,Guo Zhiqiang,Li Zongqiang,Li Qianru,Liu Su,Wang Yu,Yu Jinghua,Huang Jiadong
A DNA triangular prism nanomachine (TPN)–based logic device for intracellular AND-gated imaging of adenosine triphosphate (ATP) has been constructed. By using i-motif sequences and ATP-binding aptamers as logic control units, the TPN logic device is qualified to respond to the acidic environment and ATP in cancer cell lysosomes. Once internalized into the lysosome, the specific acidic microenvironment in lysosome causes the i-motif sequence to fold into a tetramer, resulting in compression of DNA tri-prism. Subsequently, the split ATP aptamer located at the tip of the collapsed triangular prism binds stably to ATP, which results in the fluorescent dyes (Cy3 and Cy5) modified at the ends of the split aptamer being in close proximity to each other, allowing Förster Resonance Energy Transfer (FRET) to occur. The FRET signals are excited at a wavelength of 543 nm and can be collected within the emission range of 646–730 nm. This enables the precise imaging of ATP within a cell. We also dynamically operate AND logic gates in living cells by modulating intracellular pH and ATP levels with the help of external drugs. Owing to the AND logic unit on TPN it can simultaneously recognize two targets and give corresponding intelligent logic judgment via imaging signal output. The accuracy of molecular diagnosis of cancer can be improved thus eliminating the false positive signal of single target-based detection. Hence, this space-controlled TPN-based logical sensing platform greatly avoids sensitivity to extracellular targets during the cell entry process, providing a useful tool for high-precision imaging of the cancer cell’s endogenous target ATP.
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