A convenient smartphone-assisted colorimetric for 6-Mercaptopurine detection using enhanced oxidase-like activity of ß-cyclodextrin modified MnO2 nanosheets.

6-mercaptopurine (6-MP) is widely used in the treatment of many diseases, but exhibits some serious side effects due to its toxicity. Therefore, it is important and imperative to effectively control and monitoring concentration of 6-MP. Herein, we designed a smartphone-assisted colorimetric sensing platform for 6-MP detection, based on an excellent ß-cyclodextrin modified MnO2 nanosheets (ß-CD@MnO2 NNS) mediated oxidase-like activity. ß-CD@MnO2 NNS can directly oxidizes 3,3',5,5'-tetramethylbenzidine (TMB) into oxidized TMB with color changes, yielding more than 3-fold higher oxidase-like catalytic activity compared with individual MnO2 NNS. After adding 6-MP, ß-CD@MnO2 NNS can be reduced to Mn2+ and lose their oxidase-like properties, resulting in a color and absorbance change for sensitive and selectivity detection of 6-MP. Meanwhile, the smartphone-based color recognition application can intuitively and simply measure the concentration of 6-MP. The limits of detection UV-vis instrument and smartphone were 0.35 µM and 0.86 µM, respectively. This method has also been successfully applied to the detection of real samples. Finally, this study provides a new promising platform for detection of 6-MP and is expected to be used in application of pharmaceutical analysis and biomedicine.

Recent advances in functional nucleic acid decorated nanomaterials for cancer imaging and therapy.

Nanomaterials possess unusual physicochemical properties including unique optical, magnetic, electronic properties, and large surface-to-volume ratio. However, nanomaterials face some challenges when they were applied in the field of biomedicine. For example, some nanomaterials suffer from the limitations such as poor selectivity and biocompatibility, low stability, and solubility. To address the above-mentioned obstacles, functional nucleic acid has been widely served as a powerful and versatile ligand for modifying nanomaterials because of their unique characteristics, such as ease of modification, excellent biocompatibility, high stability, predictable intermolecular interaction and recognition ability. The functionally integrating functional nucleic acid with nanomaterials has produced various kinds of nanocomposites and recent advances in applications of functional nucleic acid decorated nanomaterials for cancer imaging and therapy were summarized in this review. Further, we offer an insight into the future challenges and perspectives of functional nucleic acid decorated nanomaterials.

Enzymatic reaction modulated DNA assembly on graphitic carbon nitride nanosheets for sensitive fluorescence detection of acetylcholinesterase activity and inhibition.

A novel fluorescent strategy has been developed by using an enzymatic reaction modulated DNA assembly on graphitic carbon nitride nanosheets (CNNS) for the detection of acetylcholinesterase (AChE) activity and its inhibitors. The two-dimensional and ultrathin-layer CNNS-material was successfully synthesized through a chemical oxidation and ultrasound exfoliation method. Because of its excellent adsorption selectivity to ssDNA over dsDNA and superior quenching ability toward the fluorophore labels, CNNS were employed to construct a sensitive fluorescence sensing platform for the detection of AChE activity and inhibition. The detection was based on enzymatic reaction modulated DNA assembly on CNNS, which involved the specific AChE-catalyzed reaction-mediated DNA/Hg2+ conformational change and subsequent signal transduction and amplification via hybridization chain reaction (HCR). Under the excitation at 485 nm, the fluorescence signal from 500 to 650 nm (λmax = 518 nm) of the developed sensing system was gradually increased with increasing concentration of AChE. The quantitative determination range of AChE is from 0.02 to 1 mU/mL and the detection limit was 0.006 mU/mL. The developed strategy was successfully applied to the assay of AChE in human serum samples, and can also be used to effectively screen AChE inhibitors, showing great promise providing a robust and effective platform for AChE-related diagnosis, drug screening, and therapy.

An "on-off-on" fluorescence probe for glyphosate detection based on Cu2+ modulated g-C3N4 nanosheets.

The analysis of glyphosate is essential to agricultural production, environment protection and public health. Herein, we proposed a fast and convenient "on-off-on" fluorescence platform for sensitive detection of glyphosate via Cu2+ modulated g-C3N4 nanosheets. The fluorescence of the system was quenched by Cu2+. With the presence of glyphosate, the fluorescence could be restored due to the formation of Cu2+- glyphosate complex. The proposed method was cost-effective with label-free and enzyme-free. Moreover, it exhibits high sensitivity with a low detection limit of 0.01 µg/ml. Furthermore, the proposed method has been successfully monitored glyphosate in real samples.

An amplified fluorescence polarization assay for sensitive sensing of organophosphorus pesticides via MnO2 nanosheets.

It is highly desirable to develop a simple, efficient and sensitive strategy for organophosphorus pesticides (OPs) in both environment pollution and human health. Herein, a novel amplified fluorescence polarization (FP) biosensor was established for highly sensitive detection of OPs using MnO2 nanosheets as the signal enhancer. In this system, OPs can suppress the activity of acetylcholinesterase (AChE) efficiently, blocking the hydrolysis reaction of acetylthiocholine (ATCh) to generate thiocholine (TCh) by AChE. TCh can lead the decomposition of MnO2 nanosheets to manganese ions. So, without the influence of TCh, MnO2 nanosheets can maintain its original shape and form a stable complex with FAM-DNA, which greatly enhanced the FP signal. This method can tremendously improve the sensitivity of FP with a detection limit of 0.01 ng/mL for diazinon. In addition, it was also applicable to determine other four OPs and investigate the level of diazinon in real water samples. Consequently, the proposed approach provides a new promising platform for detection of OPs and is expected to be used in application of environmental monitoring.

A unimolecular DNA fluorescent probe for determination of copper ions based on click chemistry.

A homogenous fluorescence method was constructed for Cu2+ detection by employing DNA-templated click chemistry and exonuclease reaction. In this strategy, a dumbbell shaped DNA probe, which contained an alkyne group and an azide group at its ends, was designed as the template for the click chemistry reaction, and also the signal probe. In the absence of Cu2+, the DNA probe was digested into small oligonucleotide fragments by exonuclease, resulting in a low fluorescence background. However, this DNA probe can be sealed at its two ends by Cu2+-induced click chemistry ligation in the presence of Cu2+. This closed structure of DNA would remain stable after addition of exonuclease, and could then be stained by SYBR Green I. A strong fluorescence signal was observed, which was related to the concentration of Cu2+. This assay showed high selectivity and reached the detection limit of 39 nM. Moreover, the proposed strategy exhibited satisfactory detection results in real complex sample analysis, and has promising application in environmental monitoring and food safety.

A sensitive fluorescence assay of organophosphorus pesticides using acetylcholinesterase and copper-catalyzed click chemistry.

Organophosphorus pesticides (OPs) are widely used in agricultural fields, but exhibit high toxicity to human beings. A sensitive fluorescence assay for organophosphorus pesticides was developed using the inhibition of acetylcholinesterase (AChE) activity and the copper-catalyzed click chemical reaction. In the click reaction, two hybridized DNA probes can be ligated with copper ions, inducing a fluorescence quenching during the strand displacement reaction. AChE can hydrolyze acetylthiocholine (ATCh) to form thiocholine (TCh) which contains a thiol group. TCh will react with copper ions, blocking the click reaction and a high fluorescence signal is observed. But in the presence of OPs, the activity of AChE is inhibited, releasing a high concentration of copper ions that catalyze the click chemical reaction and resulting in decreased fluorescence signals. Taking advantage of the copper-mediated signal amplification effect, the sensitivity was improved. This assay has also been applied to detect OPs in river water samples with satisfactory results, which demonstrates that the method has great potential for practical applications in environmental protection and food safety fields.

Ultrasensitive fluorescent detection of nucleic acids based on label-free enzymatic-assisted cascade signal amplification.

A simple and homogenous label-free fluorescent method based on target-triggered and enzymatic-assisted isothermal cascade signal amplification was established for nucleic acid detection. This amplification method consists of two circuits that depend on polymerase and nicking endonuclease. In the presence of a target, a hairpin probe was opened to initiate the polymerization and nicking reaction, producing target analogues and G-rich sequences, and releasing the original target DNA. The released target and produced target analogues then trigger a new amplification cycle. Large amounts of G-rich sequences were generated through this cascade amplification process. The fluorogenic dye thioflavin T (ThT) specifically recognized G-rich sequences to form a G-quadruplex/ThT complex, which induced a strong fluorescence intensity. The approach was ultrasensitive for nucleic acid detection, and the detection limit was as low as 5.6 fM. The system discriminates single-nucleotide mutations in DNA and provides a promising strategy for nucleic acid detection in complex biological samples. This simple, label-free and ultrasensitive approach is a promising tool for biomedical research and clinical diagnostics.

A novel multiplex signal amplification strategy based on microchip electrophoresis platform for the improved separation and detection of microRNAs.

A multiplex fluorescence signal amplification method based on microchip electrophoresis (MCE) platform was developed for the improvements in the separation and detection of microRNAs. The method used two kinds of fluorescein amidite labeled DNA signal probes to hybridize with its target microRNAs, utilizing T7 exonuclease assisting target circling realized the fluorescence signal amplification. Then, two kinds of fluorescein amidite labeled DNA segments with different size were separated and detected on the MCE-laser induced fluorescence detection platform. The microRNAs-126 and microRNAs-141 were used as model analytes in the proof-of-concept experiments. Two calibration curves between the fluorescence intensity and microRNAs concentration all showed good linearity in the range of 0.025-20 nM. The correlation coefficients obtained were 0.9975 and 0.9925, respectively. The limits of detection for two kinds of microRNAs were estimated to all be 15 pM. By spiking T24 cell lysate samples with varying amounts of miRNA-126 and miRNA-141, the recovery of analytes ranged from 96.0% to 115%, and the relative standard deviations are lower than 5.5%. The present method showed high sensitivity and selectivity, which has a promising application in biomedical research.

A microchip electrophoresis-based fluorescence signal amplification strategy for highly sensitive detection of biomolecules.

We have developed a microchip electrophoresis (MCE)-based fluorescence signal amplification strategy as a universal MCE method for the detection of trace biomolecules. This strategy exhibits high sensitivity and specificity for target molecules, and has been applied for the detection of interferon-gamma (IFN-γ) in human plasma with satisfactory results.

A sensitive fluorescence turn-on assay of bleomycin and nuclease using WS2 nanosheet as an effective sensing platform.

As an important antitumor drug, bleomycin (BLM) is widely used in the treatment of a variety of cancers. In addition, nucleases play a crucial role in DNA replication, recombination and repair which are associated with cancer development. Thus, the development of BLM and nuclease detection methods is of great significance in cancer therapy and related biological mechanism research. Here, a WS2 nanosheet-based turn-on fluorescent sensing platform for simple, fast and sensitive detection of BLM and nuclease was reported. WS2 nanosheet exhibits different affinity toward ssDNA with different length and excellent fluorescence quenching ability. A fluorescein (FAM)-labeled long ssDNA could be adsorbed on the surface of WS2 nanosheet and the fluorescence was therefore quenched. In the presence of BLM·Fe(II) or S1 nuclease (a ssDNA-specific nuclease which was used as a model enzyme), an irreversible scission of long ssDNA was underwent through the BLM-induced oxidation cleavage or S1 nuclease-induced enzymatic hydrolysis. Short FAM-linked oligonucleotide fragments which could not be adsorbed on the nanosheet surface were then produced, resulting in a weak fluorescence quenching after mixing WS2 nanosheets. Thus, the fluorescence signal was restored. The proposed sensor displays a wide linear range and a high sensitivity with a detection limit of 0.3 nM for BLM and 0.01 U mL(-1) for S1 nuclease. It also exhibits a good performance in complex biological samples. This method not only provides a strategy for BLM or S1 nuclease assay but also offers a potential application in biomedical and clinical study.

A convenient label free colorimetric assay for pyrophosphatase activity based on a pyrophosphate-inhibited Cu(2+)-ABTS-H2O2 reaction.

The evaluation of pyrophosphatase (PPase) activity plays an important role in diagnosing diseases and understanding the function of PPase in the related biological events. In this work, an inhibition effect of pyrophosphate (PPi) on Cu(2+)-catalyzed H2O2-mediated oxidation of 2,2-azinobis(3-ethylbenzothiazoline)-6-sulfonic acid (ABTS) was observed. We utilize this inhibition effect to develop a convenient label free visual method for PPase activity detection. A hydroxyl radical could be generated from a Cu(2+)-based Fenton-like reaction, and then reacted with ABTS to produce colored ABTS˙(+). The strong complexation between PPi and Cu(2+) disturbed this Cu(2+)-catalyzed ABTS-H2O2 reaction probably due to changing redox potentials of Cu(2+) towards H2O2. The PPase-catalyzed hydrolysis of PPi into Pi prohibited the complexation, resulting in the recovery of catalytic capability of Cu(2+). As a result, the solution color changed from colorless to green with a remarkable increase of absorbance. Compared with the traditional PPase assays, the developed visual assay is cost-effective and easy to implement. And a high sensitivity for PPase with a detection limit of 0.027 U mL(-1) was achieved. Moreover, the proposed colorimetric strategy was also applied to evaluate PPase inhibition and exhibited a good assay performance in complex biological samples.