An improved gold nanoparticle probe-based assay for HCV core antigen ultrasensitive detection.

A gold nanoparticle probe-based assay (GNPA) was developed for ultrasensitive detection of Hepatitis C virus (HCV) core antigen. In the GNPA, after anti-HCV core antigen polyclonal antibodies and single-stranded barcode signal DNA were labeled on gold nanoparticle probe (NP), DNA enzyme was used to degrade the unbound barcode DNAs. The anti-HCV core antigen monoclonal antibodies were coated on magnetic microparticles probe (MMP).

Establishment of evanescent wave fiber-optic immunosensor method for detection bluetongue virus.

The evanescent wave fiber immunosensors (EWFI) technique was developed for the real-time rapidly sensitive and specific detection of the monoclonal antibody 3E2 of BTV. The outer-core protein VP7 of BTV was labled on the surface of the exposed fiber-optic core. The monoclonal antibody 3E2 of BTV VP7 were added and then the goat ant-rat IgG conjugated with Cy3 was captured.

Evaluation of a novel ultra-sensitive nanoparticle probe-based assay for ricin detection.

A gold nanoparticle (GNP) probe-based assay (GNPA) modified from the bio-barcode assay (BCA) was developed for ultrasensitive detection of ricin, a potential biothreat agent. In the GNPA, a chain of ricin was captured by a GNP probe coated with polyclonal antibodies and single-stranded signal DNA. A magnetic microparticle (MMP) probe coated with ricin A chain monoclonal antibody was then added to form an immuno-complex.

Development of a highly sensitive gold nanoparticle probe-based assay for bluetongue virus detection.

A simple gold nanoparticle (GNP) probe based assay (GNPA) that was modified from a bio-barcode assay (BCA) technique, was developed for ultra-sensitive, rapid detection of the bluetongue virus (BTV) VP7 outer-core protein. This assay captures the VP7 target antigen using the GNP probe coated with anti-VP7 polyclonal antibodies and single-stranded signal DNA. Magnetic microparticle (MMP) probes coated with anti-VP7 monoclonal antibodies were then added to form a sandwich immuno-complex.

Noninvasive molecular imaging of interferon beta activation in mouse liver.

Background/aims: Interferon beta (IFN-β) is the priming cytokine in the interferons (IFNs) response that plays essential roles in innate immune system. Only very few studies on IFN activation in animals have been reported before, therefore, we embarked to develop a novel method to dynamically examine IFN-β activation in mouse liver by noninvasive molecular imaging.

Methods: Interferon beta promoter-directed firefly luciferase gene was integrated into chromosomes of hepatocytes by hydrodynamic injection.

A nanoparticle-based bio-barcode assay for ultrasensitive detection of ricin toxin.

The ultrasensitive bio-barcode amplification assay (BCA) technique was developed for the specific detection of the A chain of ricin toxin. The target antigen A chain was first captured by gold nanoparticles (GNPs) coated with polyclonal antibodies. Magnetic microparticles (MMPs) coated with A chain monoclonal antibody were then added to form a sandwich immuno-complex.

Nanoparticle-based bio-barcode assay for the detection of bluetongue virus.

The ultrasensitive bio-barcode amplification assay (BCA) technique was developed for the specific detection of the outer-core protein VP7 of bluetongue virus (BTV). The target antigen VP7 was first captured by gold nanoparticles (GNPs) coated with polyclonal antibodies. Magnetic microparticles (MMP) coated with VP7 monoclonal antibody were then added to form a sandwich immuno-complex.

Detection and quantitation of bluetongue virus serotypes by a TaqMan probe-based real-time RT-PCR and differentiation from epizootic hemorrhagic disease virus.

Twenty-four serotypes of bluetongue virus (BTV) have been recognized world wide. Reliable and quantitative assays of virus universal detection are essential for fighting against BT. A real-time reverse transcription-polymerase chain reaction (RT-PCR) with a TaqMan fluorescence probe has been developed for detection of the NS1 gene of different BTV serotypes.

[The construction of reporter plasmid of ULBP1 and preliminary studying on the influence of NS3/4A on transcription of ULBP1].

Aim: To construct a report vector of ULBP1 promoter gene and preliminary study on the influence of NS3/4A on transcription of ULBP1.

Methods: ULBP1 core promoter sequence was amplified by PCR, and inserted into pGL3-enhance vector, constructing ULBP1 reporter plasmid pGL3-ULBP1; The HCV protease NS3/4A gene was subcloned into pcDNA3-Flag vector (pcDNA3-Flag-NS3/4A), and the expression of this plasmid was demonstrated by Western blot. The influence of inhibition by NS3/4A on the level of ULBP1 transcription was tested by assaying the Luciferase activity in cells transfected with pGL3-ULBP1 with Luminometer.

[Epidemiological survey on the infection of hepatitis E virus among pigs in Henan province].

Objective: To investigate hepatitis E virus (HEV) infection among pigs in Henan province.

Methods: A total of 623 swine sera, collected from 5 districts, were divided into two groups, under 3-month of age and over 3-month of age. They were tested for HEV antigen and antibody by using ELISAs, respectively.

[Secretable eukaryotic expression of recombinant chicken Iglambda and preparation of monoclonal antibodies against chicken Iglambda].

Aim: To construct the eukaryotic expression vector for chicken Iglambda light chain, to express it on COS7 cells and to prepare the monoclonal antibodies against chicken Iglambda.

Methods: The cDNA of chicken Iglambda light chain with signal peptide sequence was amplified and then inserted into eukaryotic expression plasmid pcDNA3 after double enzyme cutting. The constructed recombinant vector was transfected into COS7 cells by lipofectamin and the secretable eukaryotic expression of chicken Iglambda light chain was verified by Western blot.

Intracellular sugars improve survival of human red blood cells cryopreserved at -80 degrees C in the presence of polyvinyl pyrrolidone and human serum albumin.

Cryopreservation with impermeable protectants has great significance on storage of human red blood cells. It has become feasible to use glycerol free cryopreservation for human red blood cells. This study focuses on the effect of intracellular trehalose or glucose on human red blood cells cryopreserved in the presence of polymer.

Changes of phosphatidylserine distribution in human red blood cells during the process of loading sugars.

The plasma membrane of red blood cells permits sugars to be loaded into the cytoplasm simply by incubation in a suitable buffer solution containing the sugar. This may provide some hope for the freeze-drying of human red blood cells. However, the effect of the loading process on red blood cells has not been fully investigated.

[Progress in the study of lyophilization of human red blood cells--review].

Now the clinical preservation methods of human red blood cells mainly include hypothermic storage (4 degrees C) and cryopreservation (-80 degrees C or -196 degrees C). The preservation time of hypothermic storage of red blood cells is relatively short and it is easy to be contaminated by microbes. Cryopreservation greatly prolongs the storage time, but it needs heavy storage equipments.

[Preparation and characterization of non-agglutinating mAbs against membrane antigen on human erythrocytes].

Aim: To prepare monoclonal antibodies (mAbs) against membrane antigens on human erythrocytes and characterize their properties.

Methods: BALB/c mice were immunized with the membrane antigens of human type O erythrocytes. The splenocytes of the immunized mice were fused with Sp2/0 myeloma cells by hybridoma technique.

[Study on the origin of Rana temporaria for quality Oviduetus Ranae].

Objective: To determine the origin of Rana temporaria for quality Oviduetus Ranae in the light of historical documents and modern researches on the classification of Rana temporaria chensinensis.

Method: Works of Chinese meteria medica of all ages, related historical documents and reports from home and abroad on researches of R. temporaria chensinensis were consulted, sorted out, analyzed and summarized.

Fluorescence microplate reader measurement of tissue susceptibility to lipid peroxidation.

This unit describes a method for the measurement of cellular membrane antioxidant capacity or susceptibility of tissue samples to lipid peroxidation using a fluorescence microplate reader. The assay is simple and has the advantage of monitoring susceptibility to lipid peroxidation in a large number of samples in real time.

Vitamin E therapy in Parkinson's disease.

Though the etiology is not well understood, late-onset Parkinson's disease (PD) appears to result from several key factors including exposure to unknown environmental toxicants, toxic endogenous compounds and genetic alterations. A plethora of scientific evidence suggest that these environmental and endogenous factors cause PD by producing mitochondrial (mito) oxidative stress and damage in the substantia nigra, leading to cell death. Thus assuming a critical role for mito oxidative stress in PD, therapies to treat or prevent PD must target these mito and protect them against oxidative damage.

Thenoyltrifluoroacetone, a potent inhibitor of carboxylesterase activity.

Thenoyltrifluoroacetone (TTFA), a conventional mitochondrial complex II inhibitor, was found to inhibit purified porcine liver carboxylesterase non-competitively with a K(i) of 0.61x10(-6)M and an IC(50) of 0.54x10(-6)M.