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Biomedical subjects

Wensheng Huang

Publications and source records attributed to Wensheng Huang.

10 recordsLinked to original sources
PubMed2006

Screening genetically modified organisms using multiplex-PCR coupled with oligonucleotide microarray.

In this research, we developed a multiplex polymerase chain reaction (multiplex-PCR) coupled with a DNA microarray system simultaneously aiming at many targets in a consecutive reaction to detect a genetically modified organism (GMO). There are a total of 20 probes for detecting a GMO in a DNA microarray which can be classified into three categories according to their purpose: the first for screening GMO from un-transgenic plants based on the common elements such as promoter, reporter and terminator genes; the second for specific gene confirmation based on the target gene sequences such as herbicide-resistance or insect-resistance genes; the third for species-specific genes which the sequences are unique for different plant species. To ensure the reliability of this method, different kinds of positive and negative controls were used in DNA microarray. Commercial GM soybean, maize, rapeseed and cotton were identified by means of this method and further confirmed by PCR analysis and sequencing. The results indicate that this method discriminates between the GMOs very quickly and in a cost-saving and more time efficient way. It can detect more than 95% of currently commercial GMO plants and the limits of detection are 0.5% for soybean and 1% for maize. This method is proved to be a new method for routine analysis of GMOs.

Animals↗
PubMed2004

"Plus-C" odorant-binding protein genes in two Drosophila species and the malaria mosquito Anopheles gambiae.

Olfaction plays a crucial role in many aspects of insect behaviour, including host selection by agricultural pests and vectors of human disease. Insect odorant-binding proteins (OBPs) are thought to function as the first step in molecular recognition and the transport of semiochemicals. The whole genome sequence of the fruit fly Drosophila melanogaster has been completed and a large number of genes have been annotated as OBPs, based on the presence of six conserved cysteine residues and a conserved spacing between the cysteines. These proteins can be divided into three distinct subgroups; those with only one six-cysteine motif, those with two such motifs and those with one motif, three extra conserved cysteines and a conserved proline immediately after the sixth cysteine. This study concentrates on the last two subgroups, referred to as 'dimer' OBPs and 'Plus-C' OBPs, respectively. We determined the tissue-specific transcript levels of all of these OBP genes of D. melanogaster using semiquantitative RT-PCR. The results showed that the expression patterns can vary within a subgroup of genes and that this technique is valuable for assessing which of the putative OBP genes are likely to be involved in Drosophila olfaction. The publicly available genomes of another fruit fly Drosophila pseudoobscura, the malaria mosquito Anopheles gambiae and the yellow fever mosquito Aedes aegypti were searched by Blast against each Plus-C OBP and dimer OBP of D. melanogaster. Related genes were found in all of the other species and the relationships of these with the D. melanogaster genes and their possible biological functions are discussed.

Amino Acid Sequence↗
PubMed2004

Revisiting the odorant-binding protein LUSH of Drosophila melanogaster: evidence for odour recognition and discrimination.

LUSH is a soluble odorant-binding protein of the fruit fly Drosophila melanogaster. Mutants not expressing this protein have been reported to lack the avoidance behaviour, exhibited by wild type flies, to high concentrations of ethanol. Very recently, the three-dimensional structure of LUSH complexed with short-chain alcohols has been resolved supporting a role for this protein in binding and detecting small alcohols. Here we report that LUSH does not bind ethanol and that wild type flies are in fact attracted by high concentrations of ethanol. We also report that LUSH binds some phthalates and that flies are repelled by such compounds. Finally, our fluorescence data, interpreted in the light of the three-dimensional structure of LUSH, indicate that the protein undergoes a major conformational change, similar to that reported for the pheromone-binding protein of Bombyx mori, but triggered, in our case, by ligand.

Animals↗
PubMed2004

Voltammetric determination of tryptophan at a single-wall carbon nanotubes modified electrode.

A single-wall carbon nanotubes (SWNT)-film coated glassy carbon electrode (GCE) was described for the determination of tryptophan. In pH 2.5 Na2HPO4-citric acid buffer, tryptophan yields a well-defined and very sensitive oxidation peak at about 1.08 V at the SWNT-film coated GCE. The oxidation peak current increases greatly and the peak potential shifts toward more negative direction at the SWNT-modified GCE in contrast to that at the bare GCE. Under optimized conditions, the oxidation peak current is proportional to the concentration of tryptophan over the range from 4 x 10(-8) to 1 x 10(-5) mol/L. The detection limit is 1 x 10(-8) mol/L at 3 min of accumulation. Using the proposed method, tryptophan in the human's blood serum samples was determined.

Calibration↗
PubMed2004

Fabrication of multi-wall carbon nanotube film on glassy carbon electrode surface and the determination of tyrosine.

A novel chemically modified electrode has been prepared on the basis of the attachment of multi-wall carbon nanotubes (MWNT) onto the surface of a glassy carbon electrode (GCE) in the presence of a hydrophobic surfactant, dihexadecyl phosphate (DHP). This MWNT film was characterized by transmission electron microscopy images (TEM) and scanning electron microscopy (SEM). The electrochemical behavior of tyrosine at the MWNT film coated GCE was examined and it is found that this MWNT-modified GCE greatly enhances the oxidation peak current of tyrosine. Effects of some important factors, including pH, scan rate and amount of modifier, on the oxidation process of tyrosine were investigated. When the signal to noise ratio (SNR) is 3, the detection limit is 1 x 10(-7) M. The low relative standard deviations of the detection of tyrosine in human morning urine (3.3%) and white wine (5.2%) suggest a good reproducibility of the modified electrode.

Adsorption↗
PubMed2003

Induction of matrix metalloproteinase-13 gene expression by TNF-alpha is mediated by MAP kinases, AP-1, and NF-kappaB transcription factors in articular chondrocytes.

Tumor necrosis factor alpha (TNF-alpha), a major proinflammatory cytokine, induces arthritic joint inflammation and resorption of cartilage by matrix metalloproteinase-13 (MMP-13). RNA for MMP-13 is increased in human arthritic femoral cartilage. Mechanisms of this induction were investigated by pretreating primary human osteoarthritic (OA) femoral head chondrocytes or chondrosarcoma cells with the potential inhibitors of TNF-alpha signal transduction and downstream target transcription factors followed by stimulation with TNF-alpha and analysis of MMP-13 RNA/protein. TNF-alpha rapidly activated phosphorylation of extracellular signal-regulated kinases (ERKs), p38, and c-jun N-terminal kinase (JNK) mitogen-activated protein (MAP) kinases in human chondrocytes. Inhibitors of ERK (U0126, PD98059, and ERK1/2 antisense phosphorothioate oligonucleotide), JNK (SB203580, SP600125, and curcumin), and p38 (SB203580 and SB202190) pathways down-regulated the TNF-stimulated expression of MMP-13. Inhibitors of the transcription factors AP-1 (nordihydroguaiaretic acid, NDGA) and NF-kappaB (curcumin, proteasome inhibitors, and Bay-11-7085) suppressed TNF-alpha-induced MMP-13 expression in primary chondrocytes and SW1353 cells. These results suggest that induction of the MMP-13 gene by TNF-alpha is mediated by ERK, p38, and JNK MAP kinases as well as AP-1 and NF-kappaB transcription factors. Blockade of TNF-alpha signaling and its target transcription factors by the approaches tested here may be beneficial for reducing cartilage breakdown by MMP-13 in arthritis.

Cartilage, Articular↗
PubMed2003

Simultaneous determination of 2-nitrophenol and 4-nitrophenol based on the multi-wall carbon nanotubes Nafion-modified electrode.

In this work, multi-wall carbon nanotubes (MWNT) were conveniently dispersed into Nafion-ethanol solution, and the MWNT-Nafion-modified glassy carbon electrode (GCE) was described for the simultaneous determination of 2-nitrophenol and 4-nitrophenol. At pH 4.0 phosphate buffer, the reduction peak currents of 2-nitrophenol (at -0.8 V) and 4-nitrophenol (at -1.0 V) increase significantly at the MWNT-Nafion-modified GCE, in comparison with that at the Nafion-modified GCE and the bare GCE. The experimental parameters, such as solution pH of phosphate buffer, accumulation potential and time, and the amounts of MWNT-Nafion onto the GCE surface, were optimized. The reduction peak currents are linear with the concentration of 2-nitrophenol from 5 x 10(-8) to 1 x 10(-5) mol L(-1) and with that of 4-nitrophenol from 1 x 10(-7) to 1 x 10(-5) mol L(-1). The detection limits after 3-min accumulation are 1 x 10(-8) mol L(-1) for 2-nitrophenol and for 4 x 10(-8) mol L(-1) for 4-nitrophenol. This modified electrode was applied to direct determination of 2-nitrophenol and 4-nitrophenol in lake water samples.

Journal Article↗
PubMed2002

Anodic stripping voltammetric determination of mercury by use of a sodium montmorillonite-modified carbon-paste electrode.

A sodium montmorillonite (SWy-2)-modified carbon-paste electrode has been examined for determination of trace levels of mercury. Because of its strong cation-exchange and adsorptive characteristics, SWy-2 greatly improves the sensitivity of determination of Hg(2+). Hg(2+) is preconcentrated and reduced on the modified electrode surface at -0.40 V and then stripped from the electrode surface during the positive potential sweep. The conditions used for determination, e.g. supporting electrolyte, pH, amount of SWy-2, accumulation potential, and accumulation time, were optimized. The peak current was linearly dependent on the concentration of mercury from 1 x 10(-9) to 5 x 10(-7) mol L(-1). The detection limit (signal-to-noise ratio=3) was 1 x 10(-10) mol L(-1) after accumulation for 6 min. When the SWy-2-modified carbon-paste electrode was used to detect mercury in water samples the average recovery was 101.11%.

Journal Article↗
PubMed2002

Tissue inhibitor of metalloproteinases-4 (TIMP-4) gene expression is increased in human osteoarthritic femoral head cartilage.

Tissue inhibitor of metalloproteinases-4 (TIMP-4), the newest member of the TIMP family, blocks the activities of several matrix metalloproteinases (MMPs) implicated in the arthritic cartilage erosion. By utilizing semi-quantitative RT-PCR, immunoblotting, and immunohistochemistry, we investigated whether the TIMP-4 gene is expressed in human non-arthritic and osteoarthritic (OA) cartilage. Directly analyzed femoral head cartilage showed TIMP-4 RNA expression in 2 of 9 non-arthritic and 12 of 14 OA patients. Femoral head cartilage from 6 of 9 OA patients had elevated TIMP-4 protein compared to the low-level expression in 3 of 8 non-arthritic controls. In most patients, there was correlation between TIMP-4 RNA and protein expression. TIMP-4 protein was also detected immunohistochemically in the upper zone of OA cartilage. The widespread TIMP-4 RNA and protein expression and augmentation in femoral OA cartilage suggests its important role in joint tissue remodeling and pathogenesis of OA. Increased TIMP levels in arthritic cartilage may not be a sufficiently effective defense against cartilage resorption by excessive multiple MMPs and aggrecanases.

Aged↗