[Study on antiangiogenesis effect of Vaccaria segetalis].

Objective: To study the inhibiting effect of Vaccaria segetalis extracts on neovascularization.

Methods: The effect of Vaccaria segetalis extracts on the proliferation, migration in vitro and tube formation on Matrigel of endothelial cell (HMEC) in vivo were examined by MTT assay and Matrigel plug assay.

Results: The proliferation and migration of HMEC were inhibited significantly by Vaccaria segetalis extracts in a dose-dependent manner (IC50 = 50 microg/mL).

Identification of binding peptides of the ADAM15 disintegrin domain using phage display.

ADAM15 plays an important role in tumour development by interacting with integrins. In this study, we investigated the target peptides of the ADAM15 disintegrin domain. First, we successfully produced the recombinant human ADAM15 disintegrin domain (RADD) that could inhibit melanoma cell adhesion by using Escherichia coli.

Anti-angiogenic activity of julibroside J8, a natural product isolated from Albizia julibrissin.

Purpose: The purpose of this study was to investigate the anti-angiogenic properties of julibroside J(8), a triterpenoid saponin isolated from Albizia julibrissin.

Methods: In the presence of juliborside J(8,) the growth of human microvascular endothelial cells (HMEC-1), four human tumor cell lines, and a normal cell line (MRC-5) was evaluated by MTT assay. The in vivo anti-angiogenic effect of julibroside J(8) was evaluated on a chorioallantoic membrane (CAM) and in transplanted colon carcinoma cells in a nude mice neovascularisation model.

Dual-label time-resolved fluoroimmunoassay for simultaneous detection of aflatoxin B1 and ochratoxin A.

A dual-label time-resolved fluoroimmunoassay (TRFIA) for simultaneous quantification of aflatoxin B1 (AFB1) and ochratoxin A (OTA) is described. For this, microtitration wells were coated with AFB1-horse radish peroxidase (HRP) and OTA-bovine serum albumin. The standards and samples were loaded on the coated plates, and diluted antibodies and Eu3+- and Sm3+-labeled IgG were then added.

Preparation and biological evaluation of a glycosylated fusion interferon directed to hepatic receptors.

The antiviral activity and biodistribution of a glycosylated fusion interferon directed to hepatic receptors were evaluated to determine whether its pharmaceutical concentration in the liver could be improved. The novel glycosylated fusion interferon, galactosyl-human serum albumin-interferon-alpha2b (G-HSA-IFN) was obtained from a long-term recombinant fusion protein (HSA-IFN) by covalent coupling with a bifunctional reagent, 2-imino-2-ethyloxymethy1-1-thiogalactose. There are about 24 thiogalactose residues in each G-HSA-IFN molecular on average.

[Improving production and bioactivity of recombinant human disintegrin domain of ADAM15 (rhADAM15) in Escherichia coli].

Objective: To enhance the production and bioactivity of recombinant human disintegrin domain of ADAM15 (A Disintegrin and Metalloproteinase 15) (rhADAM15) in Escherichia coli.

Methods: The expression host was chose and the recombinant expression plasmid pGEX-ADAM15 was constructed, based on analysis of the cDNA sequence of rhADAM15.

Results: (1) 298 mg/L GST (Glutathione-S-transferase)-ADAM15 and 42 mg/L rhADAM15 were achieved when choosing E.

Screening cellular proteins involved in the anti-proliferative effect of the ADAM15 disintegrin domain in murine melanoma cells.

A disintegrin and metalloprotease protein 15 (ADAM15), a membrane-anchored glycoprotein, is believed to function in cell-cell interactions via an integrin binding motif within its disintegrin domain. To screen its target proteins, the recombinant ADAM15 disintegrin domain (RADD) was expressed in Escherichia coli, biotinylated and used in a protein pull-down assay in vitro. A total of eight kinds of proteins were identified by 2DE/LC-MS-MS.

Enhancement of recombinant human ADAM15 disintegrin domain expression level by releasing the rare codons and amino acids restriction.

This study was aimed at increasing the production of the recombinant human ADAM15 disintegrin domain (RADD) in Escherichia coli by releasing the rare codons and restricting amino acid residues. Three different strategies for increasing RADD production were examined: to select the suitable host strain, to optimize the rare codons, and to delete the amino acids residues. The total fusion protein glutathione-S-transferase (GST)-RADD concentration of 298 mg/l and 326 mg/l were achieved by selecting E.

Expression, purification, and characterization of recombinant human serum albumin fusion protein with two human glucagon-like peptide-1 mutants in Pichia pastoris.

Glucagon-like peptide-1 (GLP-1) is a 30-residue peptide hormone secreted by intestinal L-cells in response to nutrient ingestion. In the present study, overlapping PCR technology was employed to construct two GLP-1 mutants (GLP-1(A2G))2 and human albumin (HSA) genes in vitro without linker. The spliced gene, (GLP-1(A2G))2-HSA, was over expressed under the control of promoter AOX1 and Mat alpha signal peptide in Pichia pastoris.

[Use of ultrasensitive time-resolved fluoroimmunoassay for rapid detection of clenbuterol hydrochloride in urine].

Objective: In order to provide a rapid and selectivity method for the determination of clenbuterol(CBL), an indirect competitive time-resolved fluoroimmunoassay (TRFIA) was developed.

Methods: Anti-CBL antibody, was raised by immunization against CBL-BSA in rabbits. CBL-OVA was coated by physical adsorption onto the microtitre plate, CBL or sample with CBL as a competitor.

[Effect and mechanism of recombinant hirudin on fibrinolysis].

Aim: To study the effect of recombinant hirudin (rH) on tPA-induced fibrinolysis and the possible mechanism of its action.

Methods: The effect of rH on thrombin-fibrin complex (Th-Fn) was detected by 99mTc labeled rH. In the in vitro clot lysis, tPA as plasminogen activator, and recalcified plasma as plasminogen resource were used to study the influence of rH on fibrinolysis by detecting TAFIa, D-Dimer and FXIII.

[Studies on active substance of anticancer effect in Polygonum cuspidatum].

Objective: To supply the scientific basis of research and development of the medicinal value of Polygonum cuspidatum.

Methods: One composition was isolated from the roots of Polygonum cuspidatum by cytotoxicity based fractionation and identified by HPLC-MS, UV scanning and IR. The inhibition and morphology of L-02, Hep G2, SHZ-888, MCF-7, MCF-7/ADM cells growth caused by this composition was determined by MTT assay and HE dyeing.