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Biomedical subjects

S Dong

Publications and source records attributed to S Dong.

At least 37 records · Page 2Linked to original sources

Electrochemiluminescence of tris(2,2'-bipyridine)ruthenium(II) immobilized in poly(p-styrenesulfonate)-silica-Triton X-100 composite thin-films.

The electrochemiluminescence (ECL) of tris(2,2'-bipyridine)ruthenium(II) [Ru(bpy)3(2+)] immobilized in poly(p-styrenesulfonate) (PSS)-silica-Triton X-100 composite films was investigated. The cooperative action of PSS, sol-gel and Triton X-100 attached Ru(bpy)3(2+) to the electrode strongly, and the presence of Triton X-100 prevented drying fractures of the sol-gel films during gelation and even on repeated wet-dry cycles. The modified electrode was used for the ECL detection of oxalate, tripropylamine (TPA) and NADH in a flow injection analysis (FIA) system with a newly designed flow cell. The detection scheme exhibited good stability, short response time and high sensitivity. Detection limits were 0.1, 0.1 and 0.5 micromol L(-1) for oxalate. TPA and NADH, respectively, and the linear concentration range extended from 0.001 to 1 mmol L(-1) for the three analytes. Applications of the flow cell in ECL and electrochemical detection, as well as the immobilization of reagents based on the cooperative action, are suggested.

Journal Article↗

Soil temperature and plant growth stage influence nitrogen uptake and amino acid concentration of apple during early spring growth.

In spring, nitrogen (N) uptake by apple roots begins about 3 weeks after bud break. We used 1-year-old 'Fuji' Malus domestica Borkh on M26 bare-root apple trees to determine whether the onset of N uptake in spring is dependent solely on the growth stage of the plant or is a function of soil temperature. Five times during early season growth, N uptake and total amino acid concentration were measured in trees growing at aboveground day/night temperatures of 23/15 degrees C and belowground temperatures of 8, 12, 16 or 20 degrees C. We used (15NH4)(15NO3) to measure total N uptake and rate of uptake and found that both were significantly influenced by both soil temperature and plant growth stage. Rate of uptake of 15N increased with increasing soil temperature and changed with plant growth stage. Before bud break, 15N was not detected in trees growing in the 8 degrees C soil treatment, whereas 15N uptake increased with increasing soil temperatures between 12 and 20 degrees C. Ten days after bud break, 15N was still not detected in trees growing in the 8 degrees C soil treatment, although total 15N uptake and uptake rate continued to increase with increasing soil temperatures between 12 and 20 degrees C. Twenty-one days after bud break, trees in all temperature treatments were able to acquire 15N from the soil, although the amount of uptake increased with increasing soil temperature. Distribution of 15N in trees changed as plants grew. Most of the 15N absorbed by trees before bud break (approximately 5% of 15N supplied per tree) remained in the roots. Forty-six days after bud break, approximately one-third of the 15N absorbed by the trees in the 12-20 degrees C soil temperature treatments remained in the roots, whereas the shank, stem and new growth contained about two-thirds of the 15N taken up by the roots. Total amino acid concentration and distribution of amino acids in trees changed with plant growth stage, but only the amino acid concentration in new growth and roots was affected by soil temperature. We conclude that a combination of low soil temperature and plant developmental stage influences the ability of apple trees to take up and use N from the soil in the spring. Thus, early fertilizer application in the spring when soil temperatures are low or when the aboveground portion of the tree is not actively growing may be ineffective in promoting N uptake.

Amino Acids↗

Flexible use of high-density oligonucleotide arrays for single-nucleotide polymorphism discovery and validation.

A method for identifying and validating single nucleotide polymorphisms (SNPs) with high-density oligonucleotide arrays without the need for locus-specific polymerase chain reactions (PCR) is described in this report. Genomic DNAs were divided into subsets with complexity of ~10 Mb by restriction enzyme digestion and gel-based fragment size resolution, ligated to a common adaptor, and amplified with one primer in a single PCR reaction. As a demonstration of this approach, a total of 124 SNPs were located in 190 kb of genomic sequences distributed across the entire human genome by hybridizing to high-density variant detection arrays (VDA). A set of independent validation experiments was conducted for these SNPs employing bead-based affinity selection followed by hybridization of the affinity-selected SNP-containing fragments to the same VDA that was used to identify the SNPs. A total of 98.7% (74/75) of these SNPs were confirmed using both DNA dideoxynucleotide sequencing and the VDA methodologies. With flexible sample preparation, high-density oligonucleotide arrays can be tailored for even larger scale genome-wide SNP discovery as well as validation.

Cell Line↗

Cadmium-induced apoptosis and phenotypic changes in mouse thymocytes.

At present cadmium (Cd)-induced immunotoxicity and the mechanisms involved have not been fully elucidated. The main objective of the present study is to explore the apoptogenic property of Cd in primary cultured mouse thymocytes and its effect on cell surface marker expression and phenotypic changes. Cd-induced thymocyte apoptosis was determined by TdT-mediated dUTP nick end labeling (TUNEL) assay, DNA content/cell cycle analysis and DNA gel electrophoresis. The results showed that Cd was able to cause apoptosis in mouse thymocytes in a time- and dose-dependent manner. Moreover, different subsets of thymocytes possessed different susceptibility to the apoptotic effect of Cd, in the order of CD8+ > CD4- CD8- (double negative cells, DN) > CD4+ CD8+ (double positive cells, DP) > CD4+. Cd treatment also altered thymocyte surface marker expression, leading to evident phenotypic changes. Such changes were characterized by a decline in DP cells and a marked decrease in CD4+/CD8+ ratio, mainly due to a significant increase in CD8+ subsets. These observations help to obtain a better understanding of the immunotoxic and immunomodulatory effects of Cd.

Animals↗

[Water quality of closed polyculture of penaeid shrimp with tilapia and constricted tagelus].

A variety of water quality parameters were monitored in enclosures for the polyculture of penaeid shrimp with tilapia and constricted tagelus. The result showed that the environment of the polyculture systems was better than that of monoculture one. The fluctuation of DO in the water of polyculture enclosures was less violent, and their lowest values were obviously higher than those in monoculture ones. The value of COD and the number of bacteria in the water of polyculture enclosures were significantly lower than those in monoculture ones(t-test, alpha < 0.05), showing that the amount of organic matter in the water of polyculture systems was less than that in monoculture one. The biomass of plankton and the concentration of chlorophyll-a in the polyculture enclosures were also lower, showing that the suppression of the filtering animals to the plankton was obvious. The accumulation of N and P in the sediment of polyculture enclosures was 39.76% and 51.26%, lower than those of monoculture ones, respectively; and the number of bacteria in the sediment was 7.63% lower. Therefore, it is obvious that closed culture system can greatly decrease the effluent to the coastal waters caused by the pond culture industry, and decrease the pollution to the coastal waters.

Animals↗

[Effect of silver carp stocking and fertilization on plankton community in enclosures in saline-alkaline ponds].

The effect of silver carp (Hypophthalmichthys molitrix) stocking and fertilization on plankton community of enclosures ecosystem in saline-alkaline ponds were studied. After stocking silver carp in enclosures, the abundance, chlorophyll-a content and primary productivity of phytoplankton increased, and the biomass of phytoplankton was mainly composed by small species, such as diatom and green algae. Moreover, Euglnophyta and Chrysophyta were also subdominant species. The biomass of zooplankton decreased with fish stocking, and that of Cladocera was greater in control than in fish-culture enclosures. Zooplankton community was dominated by larger species such as Daphnia carinata in control enclosures. Fertilization, especially applying inorganic fertilizer, significantly increased the abundance and primary productivity of phytoplankton and the biomass of zooplankton. Although the biomass of plankton in the enclosures applied organic fertilizer was greater than that of control enclosures, the primary productivity and chlorophyll-a content of phytoplankton, the diversity index of plankton, and the P/R ratio were very low, and thus, silver carp grew slowly. The effect of filter feeding fish and applying fertilizer on the structure of plankton community was also discussed.

Animals↗

[Expression and significance of urokinase-type plasminogen activator, its receptor, and type 2 inhibitor in giant cell tumor of bone].

OBJECTIVE: To investigate the expression and significance of u-PA, u-PAR and PAI-2 mRNA in giant cell tumor of bone. METHODS: The expressions of u-PA, u-PAR and PAI-2 mRNA in 42 patients with giant cell tumor(GCT) of bone was detected with in situ hybridization. RESULTS: The expression rates of u-PA, u-PAR, and PAI-2 mRNA of multinuclear giant cells (MGC) were 64.3%, 71.4% and 40.5%. Their expression rates in the mononuclear cells (MC) were 54.8%, 45.2% and 9.5%, respectively. The expression rates of u-PA mRNA in MGC, u-PA and u-PAR mRNA in MC of grade II and III lesions were significantly higher than those of grade I and non-recurrent lesions (P < 0.05). Close positive relation was observed between the expressions of u-PA mRNA of both cells, between the expressions of u-PAR and PAI-2 mRNA in MGC and also between the expressions of u-PA and u-PAR mRNA in MC. The expression rate in MGC was higher than that of MC significantly (P < 0.05). CONCLUSION: The expressions of u-PA, u-PAR mRNA of giant cell tumor, especially in the mononuclear cells demonstrate lower differentiation and recurrent potential in giant cell tumor of bone. Mononuclear cells may be the main tumor cell in giant cell tumor of bone.

Bone Neoplasms↗

[Establishment and identification of squamous cell lines carcinoma of tongue transferred with anti-sense cyclin A gene].

OBJECTIVE: The aim of this study is to investigate the relationship between cyclin A and cycle regulation of squamous cells carcinoma of tongue, and to provide basis for cancer gene therapy. METHODS: Eukaryocyte expression vector (pAS-A) containing anti-sense and the full-length human cyclin A complementary DNA (cDNA) (1.77 kb) was constructed and was transferred into squamous carcinoma of the tongue cell line (Tca8113) by Lipofect AMINETM introduction. The positive cell clones were selected with G418. Transcription of Neo gene mRNA and cyclin A mRNA were determined by in situ hybridization. The stable expression of anti-sense cyclin A in the Tca8113 cell line was determined using immunohistochemical methods. RESULTS: After G418 selection, cells transferred anti-sense cyclin A were obtained successfully. The positive cells of in situ hybridization of specific-stained Neoprobe were observed in cells transferred eukaryocyte expression vector. The positive cells of in situ hybridization of cyclin A in cells transferred anti-sense cyclin A were significantly fewer than those in cells transferred cyclin A. The positive rate of cyclin A immunohistochemical stain in cells transferred anti-sense cyclin A was significantly lower than that in cells transferred cyclin A. CONCLUSION: Human anti-sense cyclin A gene is stably expressed in the Tca8113 cell lines.

Carcinoma, Squamous Cell↗

Electrochemiluminescent detection of chlorpromazine by selective preconcentration at a lauric acid-modified carbon paste electrode using tris(2,2'-bipyridine)ruthenium(II).

A new detection scheme for the determination of adsorbable coreactants of Ru(bpy)3(2+) electrochemiluminescent reaction is presented. It is based on selective preconcentration of coreactant onto an electrode, followed by Ru(bpy)3(2+) electrochemiluminescent detection. The coreactant employed is chlorpromazine. It was sensitively detected after 5-min preconcentration onto a lauric acid-modified carbon paste electrode. The linear concentration range was found to occur from 1 x 10(-8) to 3 x 10(-6) mol L-1 with a detection limit of 3.1 x 10(-9) mol L-1. The total analysis time is less than 10 min. As a result of selective preconcentration and medium exchange, such remarkable selectivity is achieved that reproducible quantitation of chlorpromazine in urine is possible.

2,2'-Dipyridyl↗

Conformational transition of DNA in electroreduction studied by in situ UV and CD thin layer spectroelectrochemistry.

Electrochemically induced three conformational transitions of calf thymus DNA from B10.4 to Z10.2-DNA and from B10.2 to B10.4 and to C-DNA in 10 mM phosphate buffer solution (pH 7.21) at glassy carbon electrode are found and studied by in situ circular dichroism (CD) thin layer spectroelectrochemistry with singular value decomposition least square (SVDLS) analysis. It indicates that the so-called B10.2 form and the C-form of DNA may be composed of B10.4 and left-A DNA and of B10.4 and right-A DNA, respectively. The irreversible electrochemical reduction of adenine and cytosine groups in the DNA molecule is studied by UV-Vis spectroelectrochemistry. Some electrochemical parameters alpha n = 0.17, E0' = -0.70 V (vs. Ag/AgCl), and the standard heterogeneous electron transfer rate constant, k0 = 1.8 x 10(-5) cm s(-1), are obtained by double logarithmic analysis and non-linear regression.

Animals↗

Voltammetric and spectroscopic studies on methyl green and cationic lipid bound to calf thymus DNA.

DNA interaction with cationic lipids promises to be a versatile and effective synthetic transfection agent. This paper presents the study on binding of a simple artificial cationic lipid, cetyltrimethylammonium bromide (CTAB), to calf thymus DNA (CT DNA) prior to the condensation process, taking methyl green (MG) as a probe. The results show that the CTAB binds to DNA through electrostatic interaction forming a hydrophobic complex, thus changing the micro-environment of duplex of DNA, so the binding state of MG and DNA is changed, and a complex CTAB-CT DNA-MG is formed. This fact suggests a new way to mediate the conformation of molecular assemblies of DNA and lipids.

Animals↗

Amperometric quantification of polar organic solvents based on a tyrosinase biosensor.

A novel amperometric biosensor for quantification of the electrochemically inert polar organic solvents based on tyrosinase electrode was preliminarily reported. The biosensor was fabricated by simply syringing an aqueous solution of tyrosinase/PVAVP (PVAVP: copolymer of poly(vinyl alcohol) grafting with 4-vinylpyridine) onto glassy carbon electrode surface followed by drying the modified electrode at +4 degrees C in a refrigerator. The current generated from electrochemical reduction of quinone is a probe signal. The biosensor can be used for quantification of polar organic solvents, and its mechanism was characterized with in situ steady-state amperometry-quartz crystal microbalance experiments. The detection limit, sensitivity, and dynamic range for certain organic solvents are dependent on the kind and concentration of the substrate probe and the hydrophobicity of the immobilization matrix. The response time for all the tested organic solvents is less than 2 min.

Acetonitriles↗

Composition, synthesis, and assembly of the embryonic chick retinal basal lamina.

To study the biology of basal laminae in the developing nervous system the protein composition of the embryonic retinal basal lamina was investigated, the site of synthesis of its proteins in the eye was determined, and basal lamina assembly was studied in vivo in two assay systems. Laminin, nidogen, agrin, collagen IV, and XVIII are major constituents of the retinal basal lamina. However, only agrin is synthesized by the retina, whereas the other matrix constituents originate from cells of the ciliary body, the lens, or the optic disc. The synthesis from extraretinal tissues infers that the retinal basal lamina proteins must be shed from their tissues of origin into the vitreous body and from there bind to receptor proteins provided by the retinal neuroepithelium. The fact that all proteins typical for the retinal basal lamina are abundant in the vitreous body and a new basal lamina is only formed when the vitreous body was directly adjacent to the retina is consistent with the contention of the vitreous body having a function in retinal basal lamina formation. Basal lamina assembly was also studied after disrupting the retinal basal lamina by intraocular injection of collagenase. The basal lamina regenerated after chasing the collagenase with Matrigel, which served as a collagenase inhibitor. The basal lamina was reconstituted within 6 h. However, the regenerated basal lamina was located deeper in the retina than normal by reconstituting along the retracted neuroepithelial endfeet demonstrating that these endfeet are the preferred site of basal lamina assembly.

Agrin↗

Transcriptional control elements and complex initiation pattern of the TATA-less bidirectional human thymidylate synthase promoter.

The nucleotide sequences that are important for transcription of the human thymidylate synthase gene were analyzed by deletion and site-directed mutagenesis of the promoter region. Deletion analyses from the 5' and 3' ends indicated the presence of multiple positive and negative elements. The promoter had approximately the same strength in the normal or inverted orientation. The region between 161 and 141 nt upstream of the translational start codon was found to be both necessary and sufficient for high-level promoter activity in both directions and was designated the essential promoter region. This region, which is highly conserved in human, mouse and rat TS promoters, contains potential binding sites for Ets, Sp1, and LSF transcription factors. Site directed mutagenesis of each of these elements led to large decreases in promoter strength. However, inactivation of potential Sp1 and E2F elements adjacent to the essential promoter region led to increases in promoter strength. The transcriptional start site pattern was analyzed by S1 nuclease protection assays of mRNA isolated from cells transiently transfected with TS minigenes. Multiple start sites were detected, most of which were between 160 and 120 nt upstream of the AUG codon.

Animals↗

Amperometric glucose biosensor based on lipid film.

A novel glucose biosensor based on cast lipid film was developed. This model of biological membrane was used to supply a biological environment on the surface of the electrode, moreover it could greatly reduce the interference and effectively exclude hydrophilic electroactive material from reaching the detecting surface. TTF was selected as a mediator because of its high electron-transfer efficiency, and it was incorporated in the lipid film firmly. Glucose oxidase was immobilized in hydrogel covered on the lipid film. The effects of pH, operating potential were explored for the optimum analytical performance by using amperometric method. The response time of the biosensor was less than 20 s, and the linear range is up to 10 mmol l(-1) (corr. coeff. 0.9932) with the detection limit of 2 x 10(-5) mol l(-1). The biosensor also exihibited good stability and reproducibility.

Biosensing Techniques↗

Silica sol-gel composite film as an encapsulation matrix for the construction of an amperometric tyrosinase-based biosensor.

An amperometric tyrosinase enzyme electrode for the determination of phenols was developed by a simple and effective immobilization method using sol-gel techniques. A grafting copolymer was introduced into sol-gel solution and the composition of the resultant organic-inorganic composite material was optimized, the tyrosinase retained its activity in the sol-gel thin film and its response to several phenol compounds was determined at 0 mV vs. Ag/AgCl (sat. KCl). The dependences of the current response on pH, oxygen level and temperature were studied, and the stability of the biosensor was also evaluated. The sensitivity of the biosensor for catechol, phenol and p-cresol was 59.6, 23.1 and 39.4 microA/mM, respectively. The enzyme electrode maintained 73% of its original activity after intermittent use for three weeks when storing in a dry state at 4 degrees C.

Biosensing Techniques↗