JOURNAL OF CHROMATOGRAPHIC SCIENCE
·
2025
·Vol. 63
·期 10
·
IF 1.3
Zheng Jun-tao,Kong Jia-he,Yang Ting,Wang Xue-ding,Guan Shao-xing,Guan Yan-ping,Huang Min,Gao Xiang,Chao Kang,Zhu Xia
Deoxythioguanosine (dTG), incorporated into DNA as DNA-TG, is considered a more relevant thiopurine metabolite than erythrocyte thioguanine nucleotides, as erythrocytes are not the drug target. We established a simple and sensitive liquid chromatography–tandem mass spectrometry method to detect dTG in human blood nucleated cell DNA and validated its clinical applicability in Chinese patients with inflammatory bowel disease to improve individualized thiopurine treatment. dTG was released from 1 μg DNA in a single hydrolysis step, separated by the X-Terra RP18 column and quantified by tandem mass spectrometry with 2′- Deoxyguanosine-13C,15N2 as the internal standard. The calibration curve covered eight concentration levels (0.125–25 ng/mL) with the correlation coefficients r2 ≥ 0.99. The intra- and inter-assay variability was achieved less than 9.9%. This method was employed in a study where a total of 125 inflammatory bowel disease patients with a median DNA-TG level of 279.7 fmol/μg DNA (interquartile range 183.5 ~ 410.6 fmol/μg DNA). Nucleoside diphosphate-linked moiety X-type motif 15 variant carriers (*2, *3, *5, *6) showed significantly higher DNA-TG/dose and DNA-TG/erythrocyte thioguanine nucleotides ratios than wild-type patients (P = 0.0023, P = 0.0001). In summary, this method was simple and stable to detect dTG in DNA, which may act as a promising biomarker predicting the toxicity and efficacy in patients treated with thiopurine.
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BIOMACROMOLECULES
·
2022
·
IF 6.978
Hao Sun,Haifeng Wu,Qiao Teng,Yuanxi Liu,Hui Wang,Zhen-Gang Wang
In enzymatic active sites, the essential functional groups are spatially arranged as a result of the enzyme three-dimensional folding, which leads to remarkable catalytic properties. We are inspired to self-assemble the polylysine peptides with guanine-rich DNA and hemin as cofactor to fabricate the peroxidase-mimicking catalytic nanomaterials. The DNA can fold into G-quadruplex to provide a supramolecular scaffold and a nucleobase for supporting and coordinating hemin, and the polylysine provides amine as distal groups to promote the H2O2 adsorption to the iron of hemin. The polylysine and DNA components synergistically accelerated the hemin-catalyzed reactions, and the complex containing ε-polylysine exhibited higher activity than α-polylysine. This activity difference is attributed to the higher pKa value and more susceptible protonation of amine of ε-polylysine than α-polylysine. The ε-polylysine/DNA/hemin had similar coordination states of hemin and conformations of the components to α-polylysine/DNA/hemin but accelerated the formation of the intermediate compound I faster than α-polylysine. Theoretical simulation reveals that the unprotonated NH2 behaved like a base catalyst, similar to His-42 residue in the natural heme pocket, while the protonated NH3+ acted as an acid, which indicated that the base catalyst on the distal side of the hemin pocket is more active than the acid. This work provides an avenue to control the distribution of the catalytic residues in an enzyme-like active site and to understand the roles of the key residues of native enzymes.
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TALANTA
·
2021
·
IF 6.057
Cai-Xia Shi,Yan-Zi Li,Qing Liu,Zeng-Ping Chen,Sha-Sha Li,Ru-Qin Yu
Biosensors based on various spectroscopic techniques discriminate the target microRNA (miRNA) from non-target ones with single nucleotide polymorphisms (SNPs) according to the differences in signal intensities which can be caused by other factors besides SNPs. As a result, they are liable to produce false positive results. Herein, we report an attempt to develop a false-positive resistance, sensitive and reliable mass spectrometric platform for miRNA detection. In the proposed platform, the qualitative and quantitative information of the target miRNA was obtained through analyzing mass spectral responses of the multiply charged ions of the residual fragments of the probe DNA produced during exonuclease III assisted signal amplification reaction using an advanced data analysis method. The proposed platform could achieve sensitive and accurate quantitative results for the target miRNA (e.g., miRNA-141) in complex medium with a detection limit of about 1 pM, and unambiguously identify non-target miRNAs with SNPs based on the length distribution patterns of residual fragments of probe DNA. The findings obtained in this study might open an avenue for the design of new miRNA detection methods based on mass spectrometry in combination with various nuclease assisted signal amplification strategies.
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MOLECULES
·
2021
·Vol. 26
·期 5
·1453
·
IF 4.412
Haoran Wang,Jianhua Wei,Hong Jiang,Ye Zhang,Caina Jiang,Xianli Ma
The use of cisplatin is severely limited by its toxic side-effects, which has spurred chemists to employ different strategies in the development of new metal-based anticancer agents. Here, three novel dehydroabietyl piperazine dithiocarbamate ruthenium (II) polypyridyl complexes (6a–6c) were synthesized as antitumor agents. Compounds6aand6cexhibited better in vitro antiproliferative activity against seven tumor cell lines than cisplatin, they displayed no evident resistance in the cisplatin-resistant cell line A549/DPP. Importantly,6aeffectively inhibited tumor growth in the T-24 xenograft mouse model in comparison with cisplatin. Gel electrophoresis assay indicated that DNA was the potential targets of6aand6c, and the upregulation of p-H2AX confirmed this result. Cell cycle arrest studies demonstrated that6aand6carrested the cell cycle at G1 phase, accompanied by the upregulation of the expression levels of the antioncogene p27 and the down-regulation of the expression levels of cyclin E. In addition,6aand6ccaused the apoptosis of tumor cells along with the upregulation of the expression of Bax, caspase-9, cytochrome c, intracellular Ca2+release, reactive oxygen species (ROS) generation and the downregulation of Bcl-2. These mechanistic study results suggested that6aand6cexerted their antitumor activity by inducing DNA damage, and consequently causing G1 stage arrest and the induction of apoptosis.Keywords:dithiocarbamate;ruthenium polypyridyl complexes;antitumor activity;DNA damage;cell cycle arrest;apoptosis
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INTERNATIONAL JOURNAL OF FOOD SCIENCE AND TECHNOLOGY
·
2021
·Vol. 56
·期 11
·6039-6051
·
IF 3.713
Ruifei Wang,Meimei Fang,Xinyi Hu,Yigang Yu,Xinglong Xiao
Summary The development of effective natural antibacterial agents is important due to the insecurity of synthetic antimicrobial agents and consumer preferences. In this study, kojic acid (KA) and tea polyphenols (TP) were found to exhibit synergistic inhibitory effect against Escherichia coli O157:H7 with a fractional inhibitory concentration index of 0.25. KA combined with TP at 25% of their minimum inhibitory concentrations (MICs) completely inactivated E. coli O157:H7 within 4 h. Subsequently, propidium iodide uptake tests, genomic DNA interaction analysis, molecular docking, field emission scanning electron microscopy and biofilm formation assay were conducted to understand the synergistic antibacterial mechanism of KA-TP. The results demonstrated that KA-TP attacked the cell membrane cooperatively, thus disturbing membrane integrity and cell structure, and KA could bind to the genomic DNA to affect the biofilm formation of E. coli O157:H7. Moreover, the contamination of E. coli O157:H7 on raw salmon fillets was eliminated to varying degrees after soaking treatments with KA and/or TP for 30 min. The depuration effect was further enhanced when KA combined with TP, and more than 2-log reduction of E. coli O157:H7 was obtained under the treatment of KA-TP at 2 MIC, suggesting that KA-TP may serve as a natural compound disinfectant for ready-to-eat aquatic products.
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JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
·
2020
·
IF 3.209
Chen Zhang,Wen-Juan Liu,Tian-Yu Wang,Ting-Ting Qian,Noor Ul Ain,Lu-Qian Wu,Xiao-Ning Wu,Xue-Ping Cai,Qi Zeng,Hong-Ping Xie
Gene transfection vector polyethyleneimine (PEI) was used as a cross-linking agent to crosslink the surface epoxidized magnetic nanoparticles and aggregate them to form a small magnetic bead (MB) with multiple nanoscale bumps on its surface (i.e. the multi-bumpy small magnetic bead, mbsMB). As there is a very low content of non-magnetic components (the cross-linking agent) in the magnetic bead, the mbsMB has an ultrahigh magnetic content of 81.95 % and a smaller particle size of 1.4 μm when compared with the usual medical MB. Such a small MB also has a strong magnetic force allowing it to reach the rapid separating ability of the commonly used larger medical MB which has 8 times its volume. The mbsMB has an obvious pH sensitivity of positive and negative surface charges and the salt-free isolation of DNA has been achieved based on the electrostatic interactions between mbsMB and DNA. This avoids the desalting of the isolated DNA as well as the effects of high salt concentration on its long chain helix structure. Whether in an acidic absorbing medium, an alkalinous desorbing one or a near neutral particle-storing one, the mbsMB will have obvious surface electrostatic charges. There is also its good suspension stability in an aqueous medium which provides a good condition for isolating of DNA suitable for efficiently adsorbing and desorbing. The as-prepared MB has a unique surface structure and some excellent properties, all suitable for adsorbing DNA. In addition, a large amount of commonly used gene transfection vector PEI can be cross-linked and bonded on the surface of mbsMB, whilst still having an excellent DNA-loading ability. In summary, the mbsMB has an ultrahigh capacity of 629.49 mg/g for DNA load.
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MICROCHIMICA ACTA
·
2019
·Vol. 186
·期 2
·1-9
·
IF 5.479
Zhang Kai,Cao Jinxuan,Wu Yongxiang,Hu Futao,Li Tianhua,Wang Ying,Gan Ning
A simple and highly sensitive fluorometric method is described for the determination of the antibiotic kanamycin (Kana) in food. Dual signal amplification is accomplished by making use of double Y-shaped aptamer DNA probes acting as a capture probes and signal amplification probes. The DNA probes were immobilized on a gold bar and on a magnetic bar, respectively. On addition of Kana, the Y-shaped aptamer probe captures Kana and then is disassembled to release two single-stranded DNAs. These trigger target recycling and HCR between the two bars simultaneously. As a result, many long duplex DNA chains are formed in the supernatant. After pulling out the bars and adding the fluorescent intercalating probe SYBR Green I, strong fluorescence (with excitation/emission peaks at 497/525 nm) is induced. The use of such double Y-shaped DNA probes obviously overcomes the unspecific signal amplification by HCR which increases selectivity and sensitivity. This is due to the fact that the hairpin of HCR is separated in being present in different arms of the Y-shaped probe. Under the optimal conditions, the assay has a limit of 0.45 pg·mL −1 for Kana. It was applied to analyze spiked milk, fish and pork samples.
The scheme represents a sensitive fluorometric aptamer-based method to detect kanamycin (Kana). It is making use of a double stirring bar-assisted dual amplification strategy with zero background. Abbreviations: apt: aptamer, AuNPs: gold nanoparticles, HCR: hybridization chain reaction.
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FOOD CHEMISTRY
·
2019
·
IF 5.399
Xiang-Xiu Chen,Zheng-Zhong Lin,Cheng-Yi Hong,Qiu-Hong Yao,Zhi-Yong Huang
A dichromatic label-free aptasensor was described for sulfadimethoxine (SDM) detection. Compared with the binding of SDM-aptamer to SDM, the higher affinity of aptamer to cDNA may result in the hybridization of dsDNA. In the presence of SDM, the aptamer specifically binds to SDM, leading to a blue color of AuNPs in deposit and fluorescence at 530 nm in supernatant after adding cDNA and SGI. With no target of SDM, AuNPs protected with the aptamer re-disperse in PBS with a red color, and no fluorescence occurs in supernatant. Based on the principle, SDM can be quantitatively detected through both fluorescent emission and AuNPs color changes with recoveries ranging from 99.2% to 102.0% for fish and from 99.5% to 100.5% for water samples. An analytical linear range of 2–300 ng mL −1 was achieved with the detection limits of 3.41 ng mL −1 for water and 4.41 ng g −1 for fish samples (3 σ , n = 9).
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MICROCHIMICA ACTA
·
2019
·Vol. 186
·期 12
·1-8
·
IF 5.479
Niu Yanyan,Luo Guiling,Xie Hui,Zhuang Yujiao,Wu Xianqun,Li Guangjiu,Sun Wei
A photoelectrochemical (PEC) aptasensor for Pb(II) detection is described. A nanocomposite consisting of CdS (2.5 μm) and TiO 2 nanoparticles (10 nm) was used as a photoactive material, and gold nanochains (Au NCs) as the support for immobilization of the Pb(II)-binding aptamer. The quercetin-copper(II) complex was further employed as the intercalator for the improvement of the photoactivity by embedding it into dsDNA. In the presence of Pb(II), a Pb(II)-stabilized G-quadruplex was formed between Pb(II) and DNA S1. This is accompanied by unwinding of the dsDNA and the release of the quercetin-copper(II) complex from the surface of the sensor. This results in a decrease of the photocurrent that drops linearly from 5.0 × 10 −12 to 1.0 × 10 −8 mol·L −1 Pb(II) concentration range with a detection limit of 1.6 × 10 −12 mol·L −1 . The method was applied to the determination of Pb(II) in various samples and gave satisfactory results.
A photoelectrochemical aptasensor was fabricated for the detection of Pb(II) based on CdS-TiO 2 nanocomposite modified indium tin oxide (ITO) electrode. Gold nanochains (AuNCs) were used as anchor to immobilize the aptamers S1 and S2 that form a double helix structure by DNA hybridization. After embedding of quercetin-copper(II) complex as intercalator and electron donor, the concentrations of Pb(II) were determined by the changes of photocurrents.
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ANALYST
·
2014
·Vol. 139
·期 22
·5858-5867
·
IF 3.906
Yizhong Shen,Shaopu Liu,Ling Kong,Xuanping Tan,Youqiu He,Jidong Yang
Although various strategies have been reported for double-stranded DNA (DNA) detection, development of a time-saving, specific, and regeneratable fluorescence sensing platform still remains a desired goal. In this study, we proposed a new DNA detection method that relies on an “off-on” switch of a regenerated fluorescence biosensor based on an electron transfer mechanism from glutathione (GSH)-capped CdTe quantum dots (QDs) to nile blue (NB). Initially, the high fluorescence of GSH-capped CdTe QDs could be effectively quenched by NB due to the binding of NB to the GSH on the surface of the QDs and the electron transfer from the photoexcited GSH-capped CdTe QDs to NB. Then, the high affinity of DNA to NB enabled the NB to be dissociated from the surface of GSH-capped CdTe QDs to form a more stable complex with DNA and suppress the electron transfer process between GSH-capped CdTe QDs and NB, thereby restoring the fluorescence of NB surface modified GSH-capped CdTe QDs (QDs–NB). In addition, we have testified the regenerability of the proposed DNA senor. The corresponding result shows that this DNA sensor is stable for two reuses. This fluorescence “off-on” signal was sensitive to the concentration of DNA in the range from 0.0092 to 25.0 μg mL−1 with a good correlation coefficient of 0.9989, and the detection limit (3σ/S) was 2.78 ng mL−1. To further investigate for perfect analysis performance, the developed biosensor was applied for the determination of DNA in human fresh serum samples with satisfactory results.
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DALTON TRANSACTIONS
·
2014
·Vol. 43
·期 24
·9216-9225
·
IF 4.097
Qiong Wu,Tianfeng Chen,Zhao Zhang,Siyan Liao,Xiaohui Wu,Jian Wu,Wenjie Mei,Yanhua Chen,Weili Wu,Lingli Zeng,Wenjie Zheng
Two arene Ru(II) complexes coordinated by 2-(3-methoxyphenyl)imidazole[4,5-f][1,10]phenanthroline, [(η6-RC6H5)Ru(m-MOPIP)Cl]Cl (R = H, 1; R = CH3, 2), have been prepared under microwave irradiation; the crystal structure of 2 exhibits a typical “piano stool” conformation, with bond angles for N1–Ru1–Cl1 86.02 (14)° and N2–Ru1–Cl1 84.51 (14)°. The Ru–C distance for the Ru atom bound to the benzene ring is about 0.2178(8) nm, and the average Ru–N distance for Ru atom to the two chelating N atoms is about 0.2092(4) nm. The evaluation of in vitro anticancer activities revealed that these synthetic Ru(II) complexes selectively inhibited the growth of HepG2 hepatocellular carcinoma cells, with low cytotoxicity toward LO2 human normal liver cells. The results demonstrated that the complexes exhibited great selectivity between human cancer and normal cells by comparing with the ligand m-MOPIP. Furthermore, complexes 1 and 2 could bind to c-myc G4 DNA in groove binding mode in promising affinity, and the insertion of the methyl groups in the arene ligand contributed to strengthen the binding affinity. This was also confirmed by molecular docking calculation and 1H NMR analysis which showed that both 1 and 2 can bind in the loop constructed by A6–G9 and G21–A25 base pairs in c-myc G4 DNA to block the replication of c-myc oligomer. Taken together, these results suggest that arene Ru(II) complexes display application potential as small molecule inhibitors of c-myc G4 DNA.
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ENVIRONMENTAL POLLUTION
·
2024
·
IF 7.6
Chen Wu,Sicheng Hang,Feng Li,Yujun Wu,Shengwei Yi,Xingang Liu,Mingjie Chen,Fei Ge,Jiang Tian,Ming Zhang,Dayi Zhang
Rice rhizosphere soil-porewater microdomains exist within an iron (Fe)-rich microoxic habitat during paddy soil flooding. However, the response mechanisms of core microflora in this habitat to Fe(II)-oxidation-mediated cadmium (Cd) and phenanthrene (Phen) remain unclear. Using gel-stabilized gradient systems to replicate the microoxic conditions in the rice rhizosphere porewater, we found that microaerophilic rhizobacteria drove Fe(II) oxidation to yield iron oxides, thereby reducing the Cd and Phen contents in the rhizosphere porewater and rice (Cd and Phen decreased by 15.9%–78.0% and 10.1%–37.4%, respectively). However, co-exposure to Cd and Phen resulted in a greater reduction in the Cd uptake and a greater increase in the Phen uptake in rice as compared to those in the Cd or Phen treatments, possibly attributing to the cation-π interactions between Cd and Phen, as well as competition between the adsorption sites on the roots. The elevation of Cd-tolerant genes and Phen-degradation genes in biogenic cell-mineral aggregates unveiled the survival strategies of rhizobacteria with respect to Cd and Phen in the microoxic habitat. Potential Cd-tolerant rhizobacteria (e.g., Pandoraea and Comamonas ) and Phen-degradation rhizobacteria (e.g., Pseudoxanthobacter ) were identified through the DNA-SIP and 16S rRNA gene amplicon sequencing. Metagenomic analysis further confirmed that these core microbes harbor Cd-tolerant, Phen-degradation, and Fe(II) oxidation genes, supporting their metabolic potential for Cd and/or Phen in the microoxic habitat of the rice rhizosphere. These findings suggest the potential mechanism and ecological significance of core rhizospheric microbial-driven Fe(II) oxidation in mitigating the bioavailability of Cd and Phen in paddy soil during flooding.
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