A colorimetric zinc(II) assay based on the use of hairpin DNAzyme recycling and a hemin/G-quadruplex lighted DNA nanoladder.

A method is described for sensitive colorimetric determination of zinc(II) ions that uses (a) a Zn(II)-responsive hairpin DNAzyme that assists target recycling, (b) hybridization chain reaction, and (c) hemin/G-quadruplex DNA nanoladder. The Zn(II)-responsive split of the hairpin DNAzyme (HD) acts as the recognition and transformation probe. Upon addition of Zn (II) and enzyme strand, a duplex is formed in the loop region of hairpin. The caged initiator sequence is subsequently liberated from the HD by the Zn(II)-selective split of the substrate strand. This cleavage induces an enzyme strand recycling for the next round of cleavage. As a result, the initiator DNA is accumulated and cross-opened H1 and H2 to start a hybridization chain reaction (HCR). The caged G-quadruplex is released after the HCR to recruit hemin to form the hemin/G-quadruplex that is inserted into the DNA nanoladder. Once formed in the DNA nanoladder, these act as catalytic labels for the ABTS, resulting a green color change. This cascade amplification strategy allows 10 nM to 100 µM of Zn(II) to be linearly quantified by colorimetry at 415 nm with a detection limit of 3.5 nM. The recoveries ranged from 97.7 to 108.3% were obtained, confirming high reliability of the method for Zn2+ analysis in lake water samples. Graphical abstractA colorimetric assay for Zn2+ using hairpin DNAzyme (HD) assistant cycle and hemin/G-quadruplex lighted nanoladders is designed. A Zn2+-responsive split of HD is designed as the recognition and transformation probe. The hemin/G-Quadruplex inserted nanoladder act as reporter for signal readout.

The ultra-sensitive visual biosensor based on thermostatic triple step functional nucleic acid cascade amplification for detecting Zn2.

Here, we present an ultra-sensitive visual biosensor based on thermostatic triple step functional nucleic acid cascade amplification for detecting Zn2+. A Zn2+-assisted cDNAzyme assay is conducted and the Zn2+ is successfully converted into nucleic acids to achieve the first circular amplification within 1 h. The cleavage products prompted Hybridization Chain Reaction (HCR), forming multiple DNA nanowire structures with branched chains. And the Strand Displacement Amplification (SDA) were further empowered by the HCR products with the addition of the amplification enzyme and the endonuclease. In just 80 min, the second and third-order signal amplifications are reached. With the addition of Hemin and chromogenic substrates, the visual signal output is achieved in 15 min based on the large amount of G-quadruplex (G4) DNAzymes. This biosensor can detect levels as low as 1.075 pM Zn2+, showing a good linear range within 10 pM-100 nM. It shows considerable potential for the Zn2+ quantitative detection.

Expression of connexin 43 and E-cadherin protein and mRNA in non-small cell lung cancers in Chinese patients.

AIM: Connexin 43 (Cx43) and E-cadherin are important biomarkers related with cancer. Their expression at protein and mRNA levels was here investigated in 50 primary lung carcinoma tissues and 20 samples of adjacent normal tissue of Chinese patients with non-small cell lung cancer (NSCLC). METHODS: Protein and mRNA expression were evaluated by ABC immunohistochemistry and RT-PCR. RESULTS: (1) The positive expression rates of Cx43 and E-cadherin protein were higher in the adjacent normal tissues than those in the primary lung carcinoma tissues; (2) the positive expression rates of Cx43 and E-cadherin protein decreased with NSCLC progression; (3) the expression of E-cadherin protein was not related with the pathological type of NSCLC; and (4) the relative quantity of the Cx43 or E-cadherin mRNA expression was correlated with the the histological type, clinical stage, cancer cell differentiation and the lymph node metastasis. CONCLUSION: The data suggested that the Cx43 and E-cadherin are reduced with NSCLC progression, and might be important biomarkers for judging the metastasis and prognosis.