Oxidized low-density lipoprotein induces inflammatory responses in cultured human mast cells via Toll-like receptor 4.

Background/aims: Oxidized low-density lipoprotein (ox-LDL) is a powerful atherogen. Toll-like receptor 4 (TLR4) has a pathophysiological role in regulating inflammatory responses and atherosclerosis. Mast cells can infiltrate into the atheromatous plaque and secrete various pro-inflammatory cytokines, which significantly amplify the atherogenic processes and promote plaque vulnerability.

Generation and application of anti-ouabain IgY antibodies.

Ouabain is a bioactive hapten and is very difficult to be accurately quantified because of the lack of useful reagents. Furthermore, where ouabain is produced in the adrenal glands has not been identified. In this study, ouabain-BSA was generated for immunizing the laying hens to generate ouabain-specific IgY antibodies in chicken eggs.

Applying the extensor digitorum reflex to neurological examination.

Aim: To determine the value of the extensor digitorum reflex in neurologic examination.

Methods: The extensor digitorum, biceps, and brachioradialis reflexes were elicited in 65 patients with hemiplegia and upper-limb paralysis and in a control group of 120 apparently healthy people. Reflexes were elicited by both conventional means and a new method for the extensor digitorum reflex.

[Expression and purification of truncated fragment of extracellular segment of sodium pump alpha3 subunit in Escherichia coli by in-fusion technology].

Objective: To construct prokaryotic expression system for expressing, purifying and identifying truncated fragment of extra-cellular segment of sodium pump alpha3 subunit with pGEX-6P-1 GST gene fusion system in Escherichia coli by in-fusion technology.

Methods: According to the conservative sequence of M1-M2 and M3-M4 extra-cellular gene fragments of sodium pump a3 subunit, which published in GenBank, a serial of primers and gene fragments was designed, and directly synthesized to fuse the above two gene fragments. The fusion gene was fused with gene-specific primers by PCR, and then fusion gene fragment was fused into the single stranded homology regions of vector pGEX-6P-1 by in-fusion cloning to construct recombinant vector pGEX-Trf-alpha3 (Truncated fragment of extracellular segment of sodium pump alpha3 subunit, Trf-alpha3).

[Binding activity of polypeptide containing human Na+, K+-ATPase alpha1 subunit M1-M2 extracellular segment].

Objective: To assess the binding activity of polypeptide containing human Na+, K+-ATPase alpha1 subunit M1-M2 extracellular segment (HES1 derivative).

Methods: HES1 derivative was synthesized by Fmoc method and purified by high-performance liquid chromatography-mass spectrometry, and its binding activity was identified by radioligand binding assay.

Results: 3H-ouabain and synthetic HES1 derivative showed some binding activity with the equilibrium dissociation constant (KD) of 24.

[Comparison between anti-ouabain egg yolk(IgY) and rabbit antibody(IgG) in enzyme-linked immunosorbent assay].

Aim: To improve specificity and accuracy of endogenous ouabain measurement assay.

Methods: Anti-ouabain polyclonal antibody egg yolk (IgY) and anti-ouabain rabbit antibody (IgG) were prepared respectively. In the presence of two kinds of antibody, then the specificity and accuracy of enzyme-linked immunosorbent assay (ELISA) were compared.

[Preparation and application of anti-ouabain IgY antibody].

Objective: To prepare highly specific anti-ouabain polyclonal antibody for detecting endogenous ouabain in tissues.

Methods: Ouabain-BSA compound was used to immunize hens, and the eggs were collected one week after the first immunization. The IgY antibodies in the egg yolk were separated and purified by PEG-6000 Method, and analyzed by 12% SDS-PAGE and enzyme-linked immunosorbent assay (ELISA) for titration.