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In recent years, sex-controlled breeding has emerged as an effective strategy to enhance the yields of economic animals with different growth characteristics, while increasing the economic benefits of aquaculture. It is known that the NF-κB pathway participates in gonadal differentiation and reproduction. Therefore, we used the large-scale loach as a research model for the present study and selected an effective inhibitor of the NF-κB signaling pathway (QNZ). This, to investigates the impacts of the NF-κB signaling pathway on gonadal differentiation during a critical period of gonad development and after maturation. Simultaneously, the sex ratio bias and the reproductive capacities of adult fish were analyzed. Our results indicated that the inhibition of the NF-κB signaling pathway influenced the expression of genes related to gonad development, regulated the gene expression related to the brain-gonad-liver axis of juvenile loaches, and finally impacted the gonadal differentiation of the large-scale loach and promoted a male-biased sex ratio. Meanwhile, high QNZ concentrations affected the reproductive abilities of adult loaches and inhibited the growth performance of offspring. Thus, our results deepened the exploration of sex control in fish and provided a certain research basis for the sustainable development of the aquaculture industry., Competing Interests: Declaration of competing interest The authors have no conflicts of interest related to this research., (Copyright © 2023 Elsevier Inc. All rights reserved.)

There is a continuing high demand to design effective sensors for the determination of heavy metal ions (HMIs) since they are hazardous to both human health and the environment. In this study, we reported a facile fluorescent sensor array for rapid discrimination of HMIs based on a single gold nanocluster (AuNC) probe. This AuNC probe was prepared by using 2-mercapto-1-methylimidazole (MMI) as a ligand and polyvinypyrrolidone (PVP) as a dispersing agent. The fluorescence emission of PVP/MMI-AuNC was observed to be closely related to the pH value of the aqueous solution, which displays yellow (λ max = 512 nm) and red (λ max = 700 nm) fluorescence at pH 12.0 and 6.0, respectively. Further experiments indicated that different HMIs can produce differential effects on the photoluminescence of PVP/MMI-AuNC and thus generate distinct fluorescent responses at 512 and 700 nm. On the basis of this phenomenon, a fluorescent sensor array based on the PVP/MMI-AuNC was then built by simply changing pH value in the sensor element. A total of seven HMIs had their unique response patterns and were successfully distinguished by hierarchical cluster analysis and linear discriminant analysis both in buffer solution and spiked water samples, achieving 100% identification accuracy. This study provides a simple and powerful fingerprinting sensing platform for multiple HMIs, showing broad application prospects in the field of environmental monitoring., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Various types of bovine serum albumin (BSA)-protected fluorescent gold nanoclusters (BSA-AuNCs) have been fabricated and applied in various fields. However, the conventional synthesis methods for BSA-AuNCs usually yield a low photoluminescence quantum yield (PLQY) in solution. In this study, we systematically examined the influences of incubation time, temperature, and pH on the formation process of BSA-AuNCs and then developed a novel strategy to synthesize BSA-AuNCs with PLQY (26%), far exceeding that of existing counterparts. Of the three important factors, pH, temperature, and time, pH plays a key role in the formation of BSA-AuNCs with different compositions and fluorescence properties. The matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) results showed that BSA-Au 20 NCs with high purity can be produced at a pH value of 10 and the correct combination of incubation temperature and reaction time. The advantages of the obtained BSA-Au 20 NCs, including small size, high PLQY, long lifetime, high purity, as well as facile modification, make them ideal candidates for luminescent probes in imaging and sensing applications., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

In this study, a colorimetric sensing assay of isoniazid based on excellent oxidase-like activity of heparin sodium stabilized platinum nanoparticles (HS-PtNPs) has been demonstrated. The newly prepared HS-PtNPs exhibit a great dispersion with an average size distribution of 4.8 ± 0.6 nm, and maintain more than 90% catalytic activity under strong acid and alkali or long-term storage conditions, indicating a robust nanomaterial with attractive potential. The HS-PtNPs show distinct oxidase-like activity with an ultrahigh affinity (Km = 0.01012 mM) for 3, 3', 5, 5'-tetramethylbenzidine (TMB). More significantly, we found that the pyridine ring of isoniazid has a strong reductive hydrazyl substitution, which can compete with TMB for the catalytic site of HS-PtNPs resulting in a colorless solution. Accordingly, a colorimetric sensing of isoniazid was fabricated. A linear relationship for isoniazid was achieved in 2.5 × 10 -6 to 2.5 × 10 -4  M (R 2  = 0.998) with a low limit of detection 1.7 × 10 -6  M (S/N = 3). Recovery experiments in drug tablets show that the standard recovery rates were 95%-103%. The quantitative detection data for isoniazid in drug tablets calculated respectively from the standard method and this method exhibited a high correlation coefficient (a slope of 0.9995), suggesting that high accuracy in isoniazid detection., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Gold nanoclusters (AuNCs) have attracted much attention as signal transducers in photoluminescence chemical/biological sensors. Herein, we employ bovine serum albumin/3-mercaptopropionic acid co-modified AuNCs as a fluorescence probe, Fe 3+ as a quencher, and pyrophosphate as an alkaline phosphatase (ALP) substrate and Fe 3+ chelator to design a novel biosensor for ALP detection, achieving a detection linear range of 0.8-16 U/L and a detection limit of 0.78 U/L. The developed method is successfully applied to the detection of ALP in human osteosarcoma cells and is shown to be suited for ALP inhibitor screening., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2019. Published by Elsevier B.V.)

Although numerous sensors have been successfully fabricated for the detection of various heavy metal ions by employing fluorescent gold nanoclusters (AuNCs) as nanoprobes, serious cross-interference often occurs when these ions coexist in samples, which results in glaring errors in quantification. In this study, glutathione-protected AuNCs (GSH-AuNCs) were synthesized and found to respond to both Cu 2+ and Hg 2+ via fluorescence suppression. Intriguingly, addition of Ag + to GSH-AuNCs could completely inhibit the quenching effect of Hg 2+ while not affecting the Cu 2+ -mediated quenching process. Ag + can combine with Au + on the surface of AuNCs to form a strong Ag + -Au + metallic bond, which disrupts the interaction between Hg 2+ and Au + and thus eliminates the corresponding quenching effect. Based on this phenomenon, a simple sensing approach for highly selective and sensitive detection of Cu 2+ in aqueous solution was developed using the GSH-AuNC/Ag + complex as a fluorescent turn-off nanoprobe. The proposed method exhibited good linearity in the concentration range 0.02-10 μM with a limit of detection of 12 nM. This assay was demonstrated to be suitable for determination of Cu 2+ in real water samples even in the presence of Hg 2+ , showing great promise as a tool for assessment of environmental security and drinking water quality., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Microfluidic paper-based analytical devices (μPADs) for detection of hydrogen peroxide and glucose have been developed. The analytical performance of colorimetric detection using the conventional starch-iodine color reaction has been significantly improved by using gelatin as the surface modifier which retains the enzyme activity in the dry filter paper strip, improves antioxidability, as well as decreases the strong background signal. Under optimal conditions, the color intensities show a good linear relationship with glucose concentration ranging from 0.5 to 5 mM and hydrogen peroxide concentration from 0.5 to 6 mM, with the detection limit of 0.05 mM and 0.1 mM, respectively. In addition, the accuracy of colorimetric sensor has been successfully assessed in detecting glucose from real human serum samples and recovery value ranges from 95.7% to 97%, which are approaching to the glucose oxidase endpoint. The new colorimetric assay exhibits high sensitivity, good selectivity, acceptable stability and reproducibility. The present approach is promising for monitoring glucose for point of care diagnostic applications, especially in regions with resource-limited settings., (Copyright © 2019 Elsevier B.V. All rights reserved.)

We demonstrated a sensitive method for detection of glutathione (GSH) based on LSPR scattering spectrum using gold core-satellite nanostructure linked by T-Hg 2+ -T base pair. The core-satellite assembly caused coupling between plasmonic nanoparticles, which inducing distinct change of LSPR peak wavelength. As the interaction between Hg 2+ and GSH, the core-satellite nanostructure would be disassembled, which accompanied with spectral blue-shift of the scattering spectrum. By using this method, GSH could be quantitatively detected, and the detection limits can reach to 0.1 µM., (Copyright © 2018 Elsevier B.V. All rights reserved.)

For study of the photocatalytic reaction kinetics in a confined microsystem, a photocatalysis microreactor integrated on a microfluidic device has been fabricated using an on-line UV/vis detector. The performance of the photocatalysis microreactor is evaluated by the photocatalytic degradation of Rhodamine B chosen as model target by using commercial titanium dioxide (Degussa P25, TiO 2 ) nanoparticles as a photocatalyst. Results show that the photocatalytic reaction occurs via the Langmuir-Hinshelwood mechanism and the photocatalysis kinetics in the confined microsystem (r = 0.359 min -1 ) is about 10 times larger than that in macrosystem (r = 0.033 min -1 ). In addition, the photocatalysis activity of the immobilized TiO 2 nanoparticles in the microreactor exhibits good stability under flowing conditions. The present microchip device offers an interesting platform for screening of photocatalysts and exploration of photocatalysis mechanisms and kinetics., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Self-assembled monolayer (SAM) has been extensively applied as ideal interface layer for construction of biosensors. Its chain length and end functional groups determine the physical and chemical properties of the modified surfaces, which will affect the performance of constructed biosensors. Herein, we studied the influence of chain length of n-alkanethiols SAMs on the immunoreaction kinetics employing attenuated total reflection surface-enhanced infrared absorption spectroscopy (ATR-SEIRAS). Antibody (rabbit immunoglobulin) is assembled on carboxyl terminated SAMs of n-alkanethiols with different chain lengths (n = 3, 6, 11, 16). The whole fabrication steps of the immunoassay can be monitored in situ by the ATR-SEIRAS. From the time-dependent SEIRA spectra, the interfacial immunoreaction kinetics between the immobilized antibody and antigen (goat anti-rabbit immunoglobulin) can be evaluated. We found that the immunoreaction became faster with increasing the chain length of SAMs. This chain length dependent kinetics might be attributed to different orientations of the assembled antibody caused by different packing densities of SAMs. The present research offers a sensing platform to evaluate immunoassay kinetics and provides fundamentals for construction of immunoassay with high performance., (Copyright © 2017 Elsevier B.V. All rights reserved.)

A novel microfluidic method for measuring liquid interfacial tension using monodisperse microbubbles generated in situ has been proposed. Instead of bulky gas supply used in traditional microfluidic devices, microbubbles are efficiently generated via water electrolysis in the devices. Since the bubble formation frequency is related to the interfacial tension of liquids used, thus, precisely measuring the interfacial tension of liquids in microfluidics can be achieved. In addition, it is found that during the microbubble formation, the electrochemical potential fluctuates regularly at controlled electrolysis current, and the fluctuating period depends on the microbubble generation rate. Therefore, the change in electrochemical potential can be directly used to monitor the bubble formation process, which avoids the use of an external optical detection system. As demonstration, the interfacial tension of isopentanol solutions with different concentrations was measured, and the results show good agreement with the ones obtained using the maximum bubble pressure method, confirming the accuracy of the present method. The proposed strategy offers a simple, low cost and accurate solution to measure the liquid interfacial tension confined in microfluidic channels. The present platform is easily constructed and facilely manipulated in common laboratories, which is expected to be widely used in microfluidic-based research and application fields., (Copyright © 2017 Elsevier B.V. All rights reserved.)

In this work, bimetallic Bi/Pt nanoparticles in bovine serum albumin biomolecular scaffold (BSA-Bi/PtNPs) were synthesized through a facile and green method. As compared with BSA-PtNPs, the BSA-Bi/PtNPs possess enhanced peroxidase-like catalytic activity. Moreover, the BSA-Bi/PtNPs are stable in harsh conditions such as high temperature, extreme pH environments, and high ionic strength, as well as in common biological matrixes. These prominent advantages enable the BSA-Bi/PtNPs to be applied to a wide range of fields. Bioassays, such as serum glucose detection, extracellular hydrogen peroxide (H 2 O 2 ) monitor, and cancer cells labeling, have been realized with satisfying results. The linear range of glucose determination was from 1 to 100 μM and the limit of detection (LOD) was 0.2 μM. The H 2 O 2 released from each MCF-7 cell after stimulation was calculated to be 2.66 × 10 -16  mol/s. By utilizing folic acid as a recognition element, tumor cell could be readily distinguished by BSA-Bi/PtNPs and the LOD for MCF-7 cell detection was 90 cells., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Herein, we reported for the first time that gold nanoparticles-catalyzed 3,3',5,5'-tetramethylbenzidine-H2O2 system can serve as an ultrasensitive colorimetric pH indicator. Gold nanoparticles acted as a catalyst and imitated the function of horseradish peroxidase. The absorbance at 450 nm of the yellow-color product in the catalytic reaction exhibited a linear fashion over the pH range of 6.40-6.60. On the basis of this property, we constructed a novel sensing platform for the determination of urea, urease, and urease inhibitor. The limit of detection for urea and urease was 5 μM and 1.8 U/L, respectively. The half-maximal inhibition value IC50 of acetohydroxamic acid was found to be 0.05 mM. Urea in human urine and urease in soil were detected with satisfied results., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Numerical simulation can provide valuable insights for complex microfluidic phenomena coupling mixing and diffusion processes. Herein, a novel finite element model (FEM) has been established to extract chemical reaction kinetics in a microfluidic flow injection analysis (micro-FIA) system using high throughput sample introduction. To reduce the computation burden, the finite element mesh generation is performed with different scales based on the different geometric sizes of micro-FIA. In order to study the contribution of chemical reaction kinetics under non-equilibrium condition, a pseudo-first-order chemical kinetics equation is adopted in the numerical simulations. The effect of reactants diffusion on reaction products is evaluated, and the results demonstrate that the Taylor dispersion plays a determining role in the micro-FIA system. In addition, the effects of flow velocity and injection volume on the reaction product are also simulated. The simulated results agree well with the ones from experiments. Although gravity driven flow is used to the numerical model in the present study, the FEM model also can be applied into the systems with other driving forces such as pressure. Therefore, the established FEM model will facilitate the understanding of reaction mechanism in micro-FIA systems and help us to optimize the manifold of micro-FIA systems., (Copyright © 2015 Elsevier B.V. All rights reserved.)

The limited number of gonad-specific and gonad-related genes that have been identified in fish represents a major obstacle in the study of fish gonad development and sex differentiation. In common carp Cyprinus carpio from China's Yellow River, the ovary and testis differ in volume and weight in adult fish of the same age. Comparing sperm, egg, and somatic cell transcripts in this carp may provide insight into the mechanisms of its gonad development and sex differentiation. In the present work, gene expression patterns in the carp ovary and testis were compared using suppression subtractive hybridization. Two bidirectional subtracted complementary DNA (cDNA) libraries were analyzed in parallel using testis or ovary as testers. Eighteen nonredundant clones were identified in the male library, including 15 known cDNAs. The expression patterns of selected genes in testis and ovary were analyzed using reverse transcriptase polymerase chain reaction. Tektin-1, GAPDS, FGFIBP, IGFBP-5, and an unknown gene from the Ccmg4 clone were observed to be expressed only in testis. GSDF, BMI1b, Wt1a, and an unknown gene from the Ccme2 clone were expressed at higher levels in testis than in ovary at sexual maturity. Thirty functional expressed sequence tags (ESTs) were identified in 43 sequenced clones in the female library, including 28 known cDNAs, one uncharacterized cDNA (EST clone), and one novel sequence. Eight identified ESTs showed significant differences in expression between the testis and the ovary. ZP3C and Psmb2 were expressed exclusively in ovary, whereas the expression levels of IFIPGL-1, Setd6, ATP-6, CDC45, AIF-1, and an unknown gene from the Ccfh2 clone were more strongly expressed in ovary than in testis. In addition, the expression of ZP3C, Wt1a, and Setd6 was analyzed in male and female gonads, heart, liver, kidney, and brain. ZP3C was expressed only in ovary. Setd6 expression was significantly stronger in female tissues than that in the male, except in the liver, and Wt1a expression showed sexual dimorphism in the kidney and liver. Results suggest that these genes could play key roles during carp growth, both in the gonad and other tissues. The results provide a resource for further investigation of molecular mechanisms responsible for gonad development and sex differentiation in Yellow River common carp., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Determination of protein concentration in mild environments is of great significance in the clinic diagnose and bioassay. Herein, a simple, fast and sensitive method for protein quantitative determination in neutral solution (pH 7.0) is developed. This assay is based on competition adsorption of the sample protein and fluorescently labeled dog serum albumin (FITC-DSA) onto gold nanoparticles (AuNPs). As the competitor FITC-DSA molecules are added into the mixture solution of sample protein conjugated AuNPs, they will compete for active sites of AuNPs, resulting in decrease in fluorescence intensity due to the quenching effect of AuNPs via fluorescence resonance energy transfer (FRET). Thus, quantitative determination of sample protein concentration can be achieved. Under the optimum conditions, the decrease in fluorescence intensity of the solution is related to the concentration of sample protein and a low detection limit of 0.01 µg/mL BSA can be achieved in 5 min. For the validation of our strategy in practical applications, the total protein content in human serum was determined using the as-proposed method. The result is in well agreement with that of measured by other conventional methods, suggesting a simple, accurate, and mild approach for protein detection in bioassay., (Copyright © 2015 Elsevier B.V. All rights reserved.)

A facile electrochemical immunosensor based on graphene-three dimensional nanostructure gold nanocomposites (G-3D Au) using simple and rapid one-step electrochemical co-reduction technique was developed for sensitive detection of topoisomerase. The resultant G-3D Au nanocomposites were characterized by scanning electron microscopy, cyclic voltammetry and electrochemical impedance spectroscopy, and then were used as a substrate for construction of the "sandwich-type" immunosensor. Amperometric current-time curve was employed to monitor the immunoreaction on the protein modified electrode. The proposed method could respond to topoisomerase with a linear calibration range from 0.5 ng mL(-1) to 50 ng mL(-1) with a detection limit of 10 pg mL(-1). This new biosensor exhibited a fast amperometric response, high sensitivity and selectivity, and was successfully used in determining the topoisomerase which was added in human serum with a relative standard deviation (n=5)<5%. The immunosensor served as a significant step toward the practical application of the immunosensor in clinical diagnosis and prognosis monitor., (Copyright © 2014 Elsevier B.V. All rights reserved.)

As a peroxidase mimic, cupric oxide nanoparticles were found to enhance the chemiluminescence (CL) of luminol-H(2)O(2) system up to 400 folds. The CL spectra and radical scavengers were conducted to investigate the possible CL enhancement mechanism. It was suggested that the enhanced CL could be attributed to the peroxidase-like activity of CuO nanoparticles, which effectively catalyzed the decomposition of hydrogen peroxide into hydroxyl radicals. The effects of the reactant concentrations and some organic compounds were also investigated. The proposed method could be used as a sensitive detection tool for hydrogen peroxide and glucose., (Copyright © 2012 Elsevier B.V. All rights reserved.)

As a label-free alternative of conventional flow cytometry, chip-based impedance measurement for single cell analysis has attracted increasing attentions in recent years. In this paper, we designed a T-shape microchannel and fabricated a pair of gold electrodes located horizontally on each side of the microchannel using a transfer printing method. Instant electric signals of flowing-through single cells were then detected by connecting the electrodes to a Keithley resistance and capacitance measurement system. Experimental results based on the simultaneous measurement of resistance and capacitance demonstrated that HL-60 and SMMC-7721 cells could be differentiated effectively. Moreover, SMMC-7721 cells at normal, apoptotic and necrotic status can also be discriminated in the flow. We discussed the possible mechanism for the discrimination of cell size and cell status by electrical analysis, and it is believed that the improvement of detection with our design results from more uniform distribution of the electric field. This microfluidic design may potentially become a promising approach for the label-free cell sorting and screening., (Copyright © 2011 Elsevier B.V. All rights reserved.)

This paper presents a simple and cost-effective UV-ablation technique for fabrication of size-tunable nanofluidics devices via photochemical decomposition reaction. UV-irradiation through a PET photomask results in continuous decomposition of poly(carbonate) (PC), forming nanochannel and carboxyl groups on the surface of the etched PC. This photochemical decomposition process occurs at molecular scale, therefore, the depth of nanochannels can be controlled at nanometer level. The etching rate is estimated to be ca. 0.015 nms(-1). To demonstrate the potential application of the present UV-ablation technique, a nanochannel was fabricated and integrated with microchannels to form a micro/nanofluidics chip for protein concentration. Using this device, about 10(3)-10(5) fold protein concentration can be achieved within 10 min. The present approach offers a simple and practical solution to fabricate nanofluidics devices at low-cost, and the resulting device could provide ideal platforms for μTAS towards various applications in biology and chemistry., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Unit operations for complicated biochemical analysis cannot usually be integrated into one substrate. A possible solution to solve this problem is to integrate multi-unit operations into two or more substrates. In this case, transporting droplets from one substrate to another is essential. In this work, a new method to transport droplets from a hydrophobic glass substrate to a piezoelectric substrate is proposed. An interdigitated transducer (IDT) and reflectors were fabricated on an optic grade 128°YX-cut lithium niobate (LiNbO(3)) substrate, and its working surface between the IDT and a reflector was modified to be hydrophobic. Droplets to be transported were first pipetted onto a glass substrate. Adjust the glass substrate so that the droplets could contact the working surface of the piezoelectric substrate, and then was moved down. These droplets could be successfully transported from the glass surface to the piezoelectric substrate because of their "adhesion work" difference. By using this mechanism, water and red dye droplets were successfully transported from glass substrate to piezoelectric substrate. As an application, droplets mixing process was demonstrated in the piezoelectric substrate by using surface acoustic wave after they have been transported from the glass substrate., (Copyright © 2011 Elsevier B.V. All rights reserved.)

The electrochemistry of a macrocyclic metal complex Fe(notpH(3)) [notpH(6)=1,4,7-triazacyclononane-1,4,7-triyl-tris(methylene-phosphonic acid)] reveals that the protonation/deprotonation of the non-coordinated P-OH groups in Fe(notpH(3)) affects its formal potential value (E(0)') considerably. Plotting E(0)' as function of solution pH gives a straight line with a slope of -585 mV pH(-1) in the pH range of 3.4-4.0, which is about ten times larger than the theoretical value of -58 mV pH(-1) for a reversible proton-coupled single-electron transfer at 20 °C. A sensitive pH responsive electrochemical switch sensor is thus developed based on Fe(notpH(3)) which shows an "on/off" switching at pH ∼ 4.0., (Copyright © 2010 Elsevier B.V. All rights reserved.)

We have successfully constructed a novel gold film with open interconnected macroporous walls of nanoparticles by combining the hydrogen bubble dynamic template synthesis with galvanic replacement reaction. After modified by a self-assembled monolayer (SAM) of 11-mercaptoundecanoic acid (MUA), the three-dimensionally (3D) interconnected macroporous Au film has been used as a biocompatible substrate for the immobilization of cytochrome c. The morphology, structure and electrochemical features of the modified and unmodified macroporous Au films were characterized by field-emission scanning electron microscopy (FESEM), energy-dispersive X-ray (EDX), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The results reveal that the resultant films had a large electroactive surface area for high protein loading, enhanced electron transfer of cytochrome c, retained electrochemical activity, good stability and repeatability. And the excellent electrochemical behaviors could be attributed to the hierarchical structure of the macroporous Au film constructed by nanoparticles., (Copyright (c) 2010 Elsevier B.V. All rights reserved.)

Three kinds of ZnO nanoparticles (NPs) with different size were characterized with transmission electron microscopy (TEM), X-ray diffraction (XRD). The potential cytotoxicity of ZnO NPs with various concentrations has been investigated using human embryonic lung fibroblasts (HELF) cells. The cytotoxicity of ZnO NPs on the normal HELF cell was evaluated by 3-(4,5-dimethylthiazol)-2,5-diphenltetrazoliumhromide (MTT) assay and characterized with photo microscopy and scanning electron microscopy (SEM). The results indicated that ZnO NPs aqueous suspensions with concentrations varying from 2.5 to 150mgL(-1) were harmful to the HELF cells and the cytotoxicity to HELF showed obviously concentration dependence. With concentrations higher than 20mgL(-1), ZnO NPs led to the livability of the tested cells lower than 10%. These results highlighted the need for caution during the use of ZnO NPs above 1.2-1.6mgL(-1) for safety. Finally, an UV-vis spectroscopic approach was developed for the determination of ZnO NPs concentration in the linear range 2-50mgL(-1) (correlation coefficients more than 0.9990), and the detection limit was 1.0mgL(-1).

Nickel hexacyanoferrate film modified gold electrode was prepared by a simple chemical deposition procedure from a fresh prepared solution containing ferricyanide, Ni(2+), and sodium nitrate. The resultant films have solo composition and are significantly stable as compared to the electrochemically deposited NiHCF films. For different concentrations of Na(+) in the solution, the formal potential values of NiHCF shift according to the Nernstian behavior with a slope of 48 mV in the range of 10(-4) to 1.0M. The NiHCF film was also used for the electrocatalytic oxidation of ascorbic acid. The anodic peak current observed in cyclic voltammetry increased with the ascorbic acid concentration. At a fixed potential under hydrodynamic conditions, the calibration plot was linear over the ascorbic acid concentration range 0.1-12 mM.

The effect of surface microstructures of microchannels on the separation efficiency in microchip electrophoresis has been studied. In order to understand the influence of the surface microstructures on the microchip electrophoresis performance, a thermostatic system was designed and fabricated for precisely control of the chip temperature since the surrounding temperature causes a significant effect on the separation efficiency. The periodically ordered microstructures (dams) of different sizes were fabricated on the microchannels by a simple printing process using different gray levels of the vector graphic software. And the influence of dam sizes in microchannels on the separation efficiency was evaluated by using the number of theoretical plates (N) and the relative resolution (Rs'). Analyzing the variations of mixing elements caused by the periodically ordered dams in the separate channel and elution time, we found that with small dam size fabricated at low gray level of ca. 30%, the resolution of dopamine and epinephrine increased slightly, while it decreased obviously with larger dam sizes especially fabricated at gray level of 50%.

Multilayered hemoglobin (Hb) molecules were successfully immobilized on three-dimensional gold film electrode modified with self-assembled monolayers (SAMs) of 3-mercaptopropylphosphonic acid. Direct electrochemistry of the immobilized multilayered Hb occurs with high thermal stability and electrochemical stability. In the multilayered Hb film, the most inner Hb molecules can directly transfer electron with the electrode, while the Hb protein beyond this layer communicates electron with the electrode via protein-protein electron exchange. In addition, the proposed functional interface can greatly enhance electron transfer rate of the immobilized Hb protein (k(s) = 15.8 +/- 2.0 s(-1)) due to the increase of roughness of the gold substrate. Under optimized experimental conditions, the multilayered Hb film displays good bioelectrocatalytic activity toward the reduction of hydrogen peroxide. This electrochemical sensor shows fast response (less than 1 s), wide linear range (7.8 x 10(-8) to 9.1 x 10(-5) M) and low detection limit (2.5 x 10(-8) M), which can be attributed to good mass transport, large Hb proteins loading per unit area and fast electron transfer rate of Hb protein.

The direct electrochemistry of cytochrome c (cyt c) on a gold electrode modified with 3-mercaptopropylphosphonic acid [HS-(CH(2))(3)-PO(3)H(2), MPPA] self-assembled monolayers (SAMs) was for the first time investigated. Electrochemical measurements and surface-enhanced infrared absorption spectroscopic reveal that the adsorption kinetics of cyt c on the MPPA-SAMs is very fast (saturation adsorption is completed within 5s) and the immobilized cyt c molecules retain their native secondary protein structure. The nature of interaction between cyt c and -PO(3)H(2) groups is mainly the electrostatic interaction. The direct electrochemistry of the immobilized cyt c on the -PO(3)H(2) terminated SAMs with short chain is nearly reversible. Its formal potential (E(0)'=18+/-3 mV vs. SCE) is very close to that of cyt c in an aqueous solution (E(0)'=18-22 mV vs. SCE). In addition, the electron transfer rate of cyt c immobilized on -PO(3)H(2) terminated SAMs is relatively slow as compared to -SO(3)H and -COOH terminated SAMs, indicating excess negative charge density on the SAMs surface will decrease the electron transfer rate of cyt c.

It is reported for the first time that the phosphonic acid terminated self-assembled layers assembled on a gold surface could be used as a functional interface to sensitively and selectively detect dopamine (DA) in the presence of highly concentrated ascorbic acid (AA). X-ray photoelectron spectroscopic (XPS) and reductive desorption measurements proved that the 3-mercaptopropylphosphonic acid [HS-(CH(2))(3)-PO(3)H(2), MPPA] was successfully immobilized on the gold surface via S-Au bond. Making use of the electrostatic interactions between the negatively charged phosphonic acid groups on the electrode surface and DA and AA, the MPPA modified electrode was successfully used to electrochemically discriminate between DA and AA. Cyclic voltammetry measurements shown that the MPPA modified electrodes almost completely inhibited the electrochemical reaction of AA. Therefore, DA can be selectively and sensitively determined by using the differential pulse voltammetry technique although high concentration of AA is present.

The electrochemical behavior of isorhamnetin (ISO) at a glassy carbon electrode was studied in a phosphate buffer solution (PBS) of pH 4.0 by cyclic voltammetry (CV) and differential pulse voltammetric method (DPV). A well-defined redox wave of ISO involving one electrons and one proton appeared. The electrode reaction is a reactant weak adsorption-controlled process with a charge transfer coefficient (alpha) of 0.586. Based on the understanding of the electrochemical process of ISO at the glassy carbon electrode, analysis of ISO can be realized. Under optimal conditions, the oxidation peak current showed linear dependence on the concentration of ISO in the range of 1.0x10(-8) to 4.0x10(-7)M and 1.0x10(-6) to 1.0x10(-5)M. The detection limit is 5.0x10(-9)M. This method has been successfully applied to the detection of ISO in tablets.

Separation and determination of dopamine and epinephrine with end-channel electrochemical (EC) detection integrated on a native printed microchip capillary electrophoresis (CE) system was investigated. Factors influencing the separation and detection were investigated and optimized. Results show manipulating EOF, which can be easily achieved by adjusting buffer pH, is a simple and effective way to achieve the baseline separation of dopamine and epinephrine in native polymeric microchips. Without surface modification of microchannel, printed microchips with advantages of low cost and easy preparation can achieve high performance like other microfluidic devices.

Generally speaking, measurement of hydroxylated radical products of salicylic acid requires a fussy separation process. In this study, we describe a simple method to electrochemically detect hydroxyl radicals (*OH) using 4-hydroxybenzoic acid (4-HBA) as the *OH trap. The *OH is generated by the Fenton reaction from iron (II) sulfate and hydrogen peroxide in a phosphate buffer solution. Experimental results show that our method can detect the OH with high sensitivity without any separation process. The differential pulse voltammetric responses show a linear dependence on the concentration of *OH in a range of 2.0x10(-6) and 1.0x10(-3)M with a determination limit down to 5.0x10(-7)M. As a demonstration, the kinetics of the Fenton reaction was mapped by measuring the reaction product of hydroxyl radical trapped by 4-HBA. The result is in good agreement with that reported previously. All the results show that the present approach could provide a simple, inexpensive and promising method for biomedicine and iatrology.

In matrix-assisted laser desorption/ionization (MALDI) Fourier transform mass spectrometry (FTMS) analyses of small oligosaccharides and amino acids, high sensitivities for oligosaccharides (10 fmol) were obtained by introducing oxidized carbon nanotubes (CNTs) with short and open-end structure as valuable matrix. The CNTs were deposited in porous anodic alumina (PAA) templates by chemical vapor deposition. Transmission electron microscopy (TEM) images show that those CNTs include low levels of amorphous carbon. Thus, the background interference signals generally caused by amorphous carbon powder in CNTs can be reduced effectively. Experiments also confirmed that the FTMS signal intensity of CNTs prepared in PAA template is much lower than that of commercial multi-wall carbon nanotubes (MCNTs). Moreover, the purified process for CNTs with mixed acid (H2SO4 and HNO3) also contributed to the minimization of background. Intense signals corresponding to alkali cation adduct of neutral carbohydrates and amino acids have been acquired. In addition, reliable quantitative analyses for urine and corn root were also achieved successfully. The present work will open a new way to the application of oxidized CNTs as an effective matrix in MALDI MS research.

A novel assay for the voltammetric detection of 18-bases DNA sequences relating to Chronic Myelogenous Leukemia (CML, Type b3a2) using methylene blue (MB) as the hybridization indicator was reported. DNA was covalently attached onto a glassy carbon electrode (GCE) through amines of the DNA bases using N-hydroxysulfosuccinimide (NHS) and N-(3-dimethylamion)propyl-N'-ethyl carbodiimidehydrochloride (EDC). The covalently immobilized single-stranded DNA (ssDNA) could selectively hybridize with its complementary DNA (cDNA) in solution to form double-stranded DNA (dsDNA) on the surface. A significant increase of the peak current for methylene blue upon the hybridization of immobilized ssDNA with cDNA in the solution was observed. This peak current change was used to monitor the recognition of CML DNA sequence. This electrochemical approach is sequence specific as indicated by the control experiments in which no peak current change was observed if a non-complementary DNA sequence was used. Factors, such as DNA target concentration and hybridization conditions determining the sensitivity of the electrochemical assay were investigated. Under optimal conditions, this sensor has a good calibration range between 1.25x10(-7) and 6.75x10(-7)M, with CML DNA sequence detection limit of 5.9x10(-8)M.

An amperometric biosensor for sensitive and selective detection of glucose has been constructed by using highly dispersed Pt nanoparticles supported on carbon nanotubes (Pt-MWCNTs) as sensing interface. The Pt-MWCNTs were synthesized by using the two-step pyrolysis method. This composite shows good electrocatalytic activity towards the oxidation of glucose in alkaline and thus can be used to selectively detect glucose. We found that detection potential and Nafion amount covered on the Pt-MWCNTs modified glassy carbon electrode had considerable influence on the selectivity for amperometric detection of glucose. Under optimal detection conditions (detection potential of 0.0V versus SCE and 10muL 1.5% Nafion), selective detection of glucose in the glucose concentration range of 1.0-26.5mM (correlation coefficient, >0.999) can be performed. The results demonstrate that the Pt-MWCNTs composite is promising for the fabrication of nonenzymatic glucose sensors.

The end of separation channel in a microchip was electrochemically mapped using the feedback imaging mode of scanning electrochemical microscopy (SECM). This method provides a convenient way for microchannel-electrode alignment in microchip capillary electrophoresis. Influence of electrode-to-channel positions on separation parameters in this capillary electrophoresis-electrochemical detection (CE-ED) was then investigated. For the trapezoid shaped microchannel, detection in the central area resulted in the best apparent separation efficiency and peak shape. In the electrode-to-channel distance ranging from 65 to 15mum, the limiting peak currents of dopamine increased with the decrease of the detection distance due to the limited diffusion and convection of the sample band. Results showed that radial position and axial distance of the detection electrode to microchannel was important for the improvement of separation parameters in CE amperometric detection.

Rapid separation and determination of acetaminophen and its hydrolysate with end-channel electrochemical (EC) detection integrated on a plastified poly(ethylene terephthalate) (PET)-toner microchip capillary electrophoresis (CE) system was investigated. In this separation and detection system, a Pt ultramicroelectrode integrated on a three-dimensional adjustor was used as working electrode. Factors influencing the separation and detection were investigated and optimized. Results show that acetaminophen and p-aminophenol can be well separated within 84s with R.S.D.<1% for migration time and R.S.D.<3.6% for detection current for both analytes. Detection limits for both analytes are determined to be 5.0muM (S/N=3). This method has been successfully applied to the detection of trace p-aminophenol in paracetamol tablets. The results demonstrate that the PET-toner microchips can obtain better performance than PDMS microfluidic devices but at much lower cost.

Detection of native amino acids was accomplished using a capillary electrophoresis (CE)-amperometric detection system, in which a single carbon fiber cylinder (CFC) working electrode instead of a carbon fiber disc (CFD) electrode was mounted in the end part of a poly(dimethylsiloxane) (PDMS)/glass hybrid microchannel. Similar to that using CFD working electrode, here, the electrochemical reduction reaction at the working electrode is also coupled from the separation high voltage system, the coupling degree is related to the in-channel length of the CFC. This property simplifies the fabrication procedure of the working electrode and also provides a convenient and sensitive means for the determination of non-electroactive ions by amperometry. The present detection mode is successfully used to electrochemically detect non-electroactive arginine (Arg), threonine (Thr), glutamic acid (Glu) and electroactive cysteine (Cys). Furthermore, by simply changing the detection potential, we can easily distinguish peak mobilities of electroactive amino acids from that of non-electroactive amino acids.

This paper presents an electrochemical detector for poly(dimethylsiloxane) (PDMS) microchip CE with a three-dimensional adjustor which makes it possible to accurately align a separate working electrode that can be easily fabricated in laboratory to the uncertain PDMS microchannel outlet. The substantial influence of the electrode-PDMS microchannel distance was investigated. The optimal electrode-outlet distance was found to be 10 microm for the PDMS microchannel with the width of 50 microm due to its relatively slow electroosmotic flow. Adrenaline and catechol were well separated, with a linear response range from 20 microM to 1 mM, and a detection limit of 2 microM for catechol, using carbon disk electrode (diameter of 300 microm). Furthermore, arginine and histidine can be well separated and detected directly in the PDMS microchannel using a Cu disk electrode (diameter of 150 microm).