Changqun Cai

Analysis of interaction between tamoxifen and ctDNA in vitro by multi-spectroscopic methods.

Multi-spectroscopic methods including resonance light scattering (RLS), ultraviolet spectra (UV), fluorescence spectra, (1)H NMR spectroscopy, coupled with thermo-denaturation experiments were firstly used to study the interaction of antitumor drug tamoxifen (TMX) with calf thymus (ctDNA) in acetate buffer solutions (pH 4.55). The interaction of TMX with ctDNA could cause a significant enhancement of RLS intensity, the hyperchromic effect, red shift of absorption spectra and the fluorescence quenching of TMX, indicating that there is an inserting interaction between TMX and ctDNA. This inference was confirmed by (1)H NMR spectroscopy. The chemical shift of the benzene proton changes significantly which indicates that TMX could insert into the base pairs of ctDNA. These studies are valuable for a better understanding the mode of TMX-ctDNA interaction further, which are important and useful for designing of new ctDNA targeted drug. And the antitumor drug TMX inserted directly into ctDNA in vitro, which can provide a lot of useful information to explore the development of new and highly effective anti-cancer drugs.

Double-Strand Displacement Biosensor and Quencher-Free Fluorescence Strategy for Rapid Detection of MicroRNA.

We describe a facile quencher-free fluorescence strategy for rapid detection of microRNAs (miRNAs) by using a novel double-strand displacement sensor. The sensor is designed with an outstanding 2-aminopurine (2-AP) fluorophore as a probe and a predesigned cDNA, which can completely complement the target miRNA and partly complement the 2-AP probe. When the target miRNA is added, the cDNA can be competed off from the cDNA\2-AP probe duplex, thereby forming a cDNA\RNA heteroduplex. The free 2-AP probe induces an increase in the fluorescent signal. A limit of detection of 5 nM and a wide linear range from 5 to 1000 nM (R(2) = 0.9971) are achieved by this assay. The rapid detection strategy can be accomplished within 2 h without expensive nanoparticles and complicated instruments for the whole procedure, thus, offering a significant potential for clinical application.

Multi-spectroscopic method study the interaction of anti-inflammatory drug ketoprofen and calf thymus DNA and its analytical application

Interactions of the anti-inflammatory drug ketoprofen with calf thymus DNA (ctDNA) in aqueous solution have been studied by multi-spectroscopic method including resonance light scattering (RLS) technique, ultraviolet spectra (UV), (1)H NMR, etc. The characteristics of RLS spectra, the effective factors and optimum conditions of the reaction have been unequivocally investigated. Mechanism investigations have shown that ketoprofen can bind to ctDNA by groove binding and form large particles, which resulted in the enhancement of RLS intensity. In Critic acid-Na(2)HPO(4) buffer (pH=6.5), ketoprofen has a maximum peak 451.5 nm and the RLS intensity is remarkably enhanced by trace amount of ctDNA due to the interaction between ketoprofen and ctDNA. The enhancement of RLS signal is directly proportional to the concentration of ctDNA in the range of 1.20×10(-6)-1.0×10(-5) mol/L, and its detection limit (3σ) is 1.33×10(-9) mol/L. The method is simple, rapid, practical and relatively free from interference generated by coexisting substance, and was applied to the determination of trace amounts of nucleic acid in synthetic samples with satisfactory results.

Interaction of anticancer drug methotrexate with nucleic acids analyzed by multi-spectroscopic method

Methotrexate (MTX) as an antifolate, which is widely used as chemotherapeutic drugs. A high-dose MTX therapy has a direct toxicity influence on the non-germinal cells, especially the liver cells. It is known that the inject dose for adults is 10-30 mg and is half for children for routine use, while our experiments showed that the optimum dosage of MTX which enhanced the RLS intensities to the maximum is 4.54 ng ml(-1). The interaction of methotrexate (MTX) with nucleic acids in aqueous solution in the presence of cetyltrimethylammonium bromide (CTMAB), a kind of cationic surfactant similar to the Human cells, were investigated based on the measurements of resonance light scattering (RLS), UV-vis, fluorescence and NMR spectra, etc. The interaction has been proved to give a ternary complex of MTX-CTMAB-DNA in BR buffer (pH 9.30), which exhibits strong enhanced RLS signals at 339.5 nm.

Y-shaped probe for convenient and label-free detection of microRNA-21 in vitro.

A simple, highly selective, and label-free microRNA (miRNA) detection method based on l-alanine-reduced graphene oxide fluorescence quenching with a Y-shaped probe is proposed. The Y-shaped probe was synthesized by silver nitrate and a cytosine-rich molecular beacon (MB) in two terminals through sodium borohydride reduction, which generated a stronger fluorescent signal than ordinary DNA-templated silver nanoclusters (AgNCs). Meanwhile, the Y-shaped probe contained a single-stranded loop structure, which could be superbly adsorbed onto the surface of reduced graphene oxide (RGO) via π-π stacking interaction, and this special structure of the probe was designed to improve its sensitivity and selectivity. In addition, the quenching capacities of graphene oxide (GO) and RGO were compared in this research. The strong interaction between nucleobases of the loop structure and RGO nanosheet made the MB-AgNCs-RGO system exhibit minimal background fluorescence. In the presence of miRNA-21, the loop structure of the Y-shaped probe can hybridize with target miRNA-21; the molecular beacon encapsulated probe is far away from RGO surface and produces a detectable signal. The MB-AgNCs based approach provides a label-free avenue to detect miRNA with high selectivity and good reproducibility, which has a promising application in early clinical diagnosis and biomedical research.

Rapid and efficient separation of glycoprotein using pH double-responsive imprinted magnetic microsphere

As biomarkers of many diseases, glycoproteins are of great significance to clinical diagnostics. However, the determination of low abundant glycoproteins in complex biological samples without any pretreatment process is still a problem. In this study, a rapid and convenient separation method for highly efficient enrichment of glycoproteins is reported, based on pH double-responsive imprinted magnetic microspheres. Thin imprinted polymer shells were fabricated onto the surface of magnetic microspheres by free radical polymerization, using 2-(Dimethylamino) ethyl methacrylate as pH-sensitive monomer, 4-vinylphenylbronic acid as boronate affinity monomer, and ovalbumin (OVA) as template molecule. Combining the advantages of pH-sensitive monomer and boronate affinity monomer, rapidly capture-release of OVA could be modulated by changing solution pH. Moreover, high absorption ability (81.2mg/g) was achieved within about 10min. This study provided responsible way to imprint glycoproteins and showed great potential for glycoprotein detection in clinical diagnostic.

A virus resonance light scattering sensor based on mussel-inspired molecularly imprinted polymers for high sensitive and high selective detection of Hepatitis A Virus

We described a novel resonance light scattering (RLS) sensor for the specific recognition of trace quantities of Hepatitis A Virus (HAV); the sensor was based on a mussel-inspired hepatitis molecularly imprinted polymer. As a recognition element, polydopamine (PDA)-coated totivirus-imprinted polymer was introduced on the surface of SiO2 nanoparticles (virus-imprinted SiO2@PDA NPs) using an efficient one-step synthesis method. The target virus was selectively captured by the imprinted polymer films, thereby increasing the RLS intensity. A simple fluorescence spectrophotometer was employed to measure the changes in the intensity. The enhanced RLS intensity (∆IRLS) was proportional to the concentration of HAV in the range of 0.04-6.0nmol∙L-1, with a low limit of detection of 8.6pmol∙L-1. The selectivity study confirmed that the resultant HAV-imprinted SiO2@PDA NPs possessed high selectivity for HAV. The sensor was successfully applied for the direct detection of additional HAV from a 20,000-fold dilution of human serum. The proposed strategy is simple, eco-friendly, highly selective, and sensitive.

A novel CdTe quantum dots probe amplified resonance light scattering signals to detect microRNA-122

We report a rapid and facile resonance light scattering (RLS) technique that utilizes CdTe quantum dots (CdTe QDs) probe to detect microRNA-122. The RLS sensor is ingeniously designed with P1 and P2, two cDNA sequence probes with partially complementary sequences to miRNA-122. The amine-modified P1 and P2 are coupled to the surface of QDs to form functional QDs-P1 and QDs-P2 conjugates, which are collectively referred to as QDs-P. The cDNAs hybridize with the target miRNA to rapidly induce the self-assembly of QDs probe and change RLS intensity. The proposed technique can detect miRNA-122 within 40min. RLS intensity is enhanced in proportion with miRNA-122 concentrations of 0.16-4.80nM and has a low detection limit of 9.4pM. In addition, the assay satisfactorily detects miRNAs in human serum samples. Thus, the assay has considerable potential for the analysis of other interesting tumor makers.

Click on the bidirectional switch: the aptasensor for simultaneous detection of lysozyme and ATP with high sensitivity and high selectivity.

A bifunctional and simple aptasensor was designed to one-spot simultaneously detect two analytes, lysozyme and ATP. The aptasensor was obtained by the electronic interaction between methyl violet (MV) and dsDNA. The dsDNA was obtained by hybridization of ATP aptamer and lysozyme aptamer. And we used the resonance light scattering (RLS) technique to detect the concentration of lysozyme and ATP. During the procedure of detection, the aptasensor works like a bidirectional switch, the corresponding side of the dsDNA will open when the target (lysozyme or ATP) “click” the aptamer, which results in corresponding RLS signal change. By the combination of the RLS technique, it is found that the changed RLS intensity was proportional to the concentration of lysozyme and ATP. The mixtures of ATP and lysozyme also met two binary function relations. The results indicated that the aptasensor could achieve simultaneous detection of ATP and lysozyme, the detection limits of ATP and lysozyme could reach 10−11 M and 10−12 M, respectively. The aptasensor shows potential application for small molecule and protein detection by RLS, it could extend the application of RLS technique.

Synthesis and Application of Molecularly Imprinted Polymer on Selective Solid-Phase Extraction for the Determination of Artemisinin in Artemisia Annua L.

Molecular imprinted polymer (MIP) was prepared through thermal polymerization by using artemisinin as the template molecule, styrene as the functional monomer, ethylene glycol dimethacrylate (EGDMA) as the cross-linker, and azobisisobutyronitrile (AIBN) as the initiator. Evaluated by static, kinetic, and bonding capacity adsorption tests, the polymer exhibited a special selective rebinding to artemisinin. Scatchard analysis revealed that two different binding sites were formed in the polymers. The novel imprinted polymer was used as a solid-phase extraction (SPE) sorbent for the extraction of artemisinin from Artemisia annua L., followed by HPLC-UV analysis. The maximum static adsorption capacity of the artemisinin-imprinted sorbent for artemisinin was 8.46 mg g−1.The application of artemisinin-imprinted polymers with high affinity and excellent selectivity toward artemisinin in SPE might offer a novel method for the enrichment and determination of the active ingredient in Chinese medicine.