Concatenated Catalytic Hairpin Assembly/Hyperbranched Hybridization Chain Reaction Based Enzyme-Free Signal Amplification for the Sensitive Photoelectrochemical Detection of Human Telomerase RNA.

Human telomerase RNA (hTR), an important biomarker for cancer diagnosis, is the template for the synthesis of telomeric DNA repeats and is found to be 7-fold overexpressed in tumor cells. Herein, we present a photoelectrochemical (PEC) biosensor for hTR detection coupled with a novel amplification strategy based on cascades of catalytic hairpin assembly (CHA) and hyperbranched hybridization chain reaction (HB-HCR). At the electrode surface, thiolated hairpin 1 probes were immobilized on deposited CdS nanoparticles via a Cd-S bond. In the presence of target hTR, a CHA reaction was triggered and the exposing of trigger1 could further initiate an HB-HCR reaction to form abundant hemin/G-quadruplex DNAzymes containing dendritic DNA structure. The DNAzymes' catalytic precipitation of 4-chloro-1-naphthol (4-CN) by H2O2 subsequently took place on the surface of the PEC electrode and efficiently suppressed the photocurrent output. Therefore, the change of photocurrent response had a positive linear relationship with logarithmic value of hTR concentration varying from 200 fM to 20.0 nM with a limit of detection (LOD) of 17.0 fM. The LOD for CHA/HB-HCR was about 8.8-fold lower than that of CHA/linear-branched HCR (CHA/LB-HCR) and 547-fold lower than that of CHA. By coupling the feature of high signal amplification capacity for DNA nanotechnology, a prominently stable, reproducible, and selective PEC biosensor was successfully constructed and applied in hTR detection.

[Preparation of mixed crystal TiO2 nanoparticles and photocatalytic degradation of toxic organic pollutants].

Mixed crystal TiO2 nanoparticles were prepared from a precursor of TiO2 by hydrothermal-steam method. The effects of the reaction temperature and the reaction time on the photocatalytic activity of the brookite TiO2 were studied. The TiO2 samples were characterized by X-ray diffraction (XRD) and transmission electron microscopy (TEM). Photocatalytic degradation of organic dye sulforhodamine B (SRB) under UV light (lambda < or = 387 nm) was used as probe reaction to evaluate the properties of the TiO2. The result showed that TiO2 prepared under 150 degrees C for 24 h had high photocatalytic activity. The size of the mixed crystal TiO2 was 14.20 nm. Brookite and anatase phase of the mixed crystal TiO2 were 63.6% and 36.4%, respectively. The dye discoloration and degradation rates were tracked, and the intermediate products hydrogen peroxide (H2O2) and hydrogen radicals (*OH) were determined during the photocatalytic experiments. The results indicated that photocatalytic degradation of brookite TiO2 mainly referred to the *OH radical mechanism. After 5 h, the mineralization and oxidation rates of SRB and 2,4-DCP mineralization were 89% and 78%, respectively. The catalyst showed good stability with no significant changes in catalytic properties after 5 cycles of SRB photocatalytic degradation experiments.

Development of a group selective molecularly imprinted polymers based solid phase extraction of malachite green from fish water and fish feed samples.

A group selective molecularly imprinted solid phase extraction (MISPE) for malachite green (MG) from fish water and fish feed samples was developed. Using MG as template molecule, methacrylic acid as functional monomer, ethylene glycoldimethacrylate as linking agent and bulk polymerization as synthetic method, the molecularly imprinted polymers (MIPs) were synthesized and characterized with rebinding experiment. The Scatchard polt's analysis revealed that the template-polymer system showed the two-site binding behavior with dissociation constants of 0.3194 micromol L(-1) and 15.70 micromol L(-1), respectively. MG and two structurally related compounds, leucomalachite green (LMG) and crystal violet (CV) were employed for selectivity test. The MIPs exhibited the highest selective rebinding to MG, but also displayed 83.0% and 87.5% of cross-reactivity with LMG and CV, demonstrating that MIPs could be used as group recognition sorbents in solid phase extraction. The extraction conditions of MISPE column for MG were optimized. Tap water samples spiked with MG at concentration of 0.5-10 ng mL(-1) were extracted by MISPE column and analyzed by high performance liquid chromatography. The recoveries of MISPE column for MG extraction were found to be 76.8-93.7% with the relative standard deviations of 2.12-10.09%, indicating the feasibility of the prepared MIPs for MG extraction. No detectable MG was observed in one fish farming water sample and two fish feed samples; while the MG concentrations in two pet fishpond water samples were found at 1.50 ng mL(-1) and 0.67 ng mL(-1), respectively.

N-(2-Hydroxy-ethyl)-3,5-dinitro-benzamide.

The title compound, C(9)H(9)N(3)O(6), was synthesized by the condensation of methyl 3,5-dinitro-benzoate and 2-amino-ethanol. The non-centrosymmetric space group results in the formation of pseudo-chiral helices in the crystal structure, which exhibits a layer packing structure involving intra-molecular N-H⋯O and O-H⋯O inter-actions.

Development and validation of a highly sensitive ELISA for the determination of pharmaceutical indomethacin in water samples.

The development and validation of a highly sensitive and specific indirect competitive enzyme-linked immunosorbent assay (ELISA) for the detection of pharmaceutical indomethacin in water samples was presented. The immunogen and coating antigen were prepared by covalently linking indomethacin to bovine serum albumin and ovalbumin by anhydride ester method. Two rabbits were immunized by standard immunization processes and the superior antibody was characterized in terms of sensitivity, specificity, precision, accuracy and stability. Under optimal experimental conditions, the standard curve was constructed in the concentration range of 0.01-10ng/mL. For 10 consecutive standard curves run in 2 weeks, IC(50) value (the concentration of analyte producing 50% of inhibition) were found within 0.10-0.25ng/mL, and the detection limit (DL) at a signal-to-noise ratio of 3 (S/N=3) was about 0.01ng/mL. The antiserum recognized acemetacin, a precursor of indomethacin with 92.3% cross-reactivity, while the cross-reactivity values of antiserum with other tested compounds were very low. From the spiking experiments, the recoveries were found within 98-123%. The ELISA was applied for the determination of indomethacin in different water samples and the results were confirmed by conventional HPLC. The correlation coefficient of 0.988 was obtained, demonstrating a good correlation of ELISA with HPLC.

Direct competitive ELISA for the determination of polychlorinated biphenyls in soil samples.

A direct competitive ELISA for determination of polychlorinated biphenyls (PCB) in soil samples was described. The standard calibration curve based on Delor 103 (Aroclor 1242) was constructed in the dynamic range of 10-1000 micro g L(-1) and a detection limit of 5.0-12.9 micro g L(-1) (or 0.5-1.29 micro g g(-1) soil) was achieved. When spiked soil samples were extracted with methanol recoveries were 90.6-106.3 %. The effect of methanol and DMSO on assay signal and sensitivity was established. Eight PCB-contaminated soil samples were analyzed by ELISA and gas chromatography (GC). The ELISA results from Soxhlet extraction were in a good agreement with those of GC (correlation coefficient 0.9866; n=8). Except for one soil sample the results from ELISA with methanol extraction were not significantly different from those from GC.