Biomedical subjects

Hongwu Du

Publications and source records attributed to Hongwu Du.

7 recordsLinked to original sources

PubMed2006

Construction and direct immunization of eukaryotic expression plasmid encoding human soluble B lymphocyte stimulator.

B lymphocyte stimulator of the tumor-necrosis-factor family (BLyS) enhances B cell survival, a function indispensable to B cell maturation. This factor plays a crucial role in enhancing immune responses. Here, to study the primary immune effect of BLyS gene in animals, we constructed recombinant eukaryotic expression plasmid of the human soluble BLyS by using expression vector pcDNA3.1 (-). The recombinant plasmid pcDNA3.1BLyS was injected subcutaneously into BALb/C mice and this administration induced the sustained expression of human BLyS and specific IgG against the recombinant BLyS protein. This study shows that recombinant eukaryotic expression plasmid, pcDNA3.1BLyS, might be used in human gene therapy.

Animals↗

PubMed2006

Development of an immunoassay kit for detecting the alteration of serum B cell activating factor in thermally injured mice.

B cell activating factor (BAFF), a member of the family of tumor necrosis factor (TNF) ligands, is essential for the development of peripheral mature, long lived B lymphocytes. Previous studies were almost related to the function or mechanism of BAFF protein and there is little report about BAFF expression in thermally injured animals. Here, we developed a special ELISA kit to study the change of BAFF expression in thermally injured mouse model. It was shown that BAFF expression changed in double-phase. Peripheral BAFF level dropped rapidly after thermal injury and at 24 h of scald it was only 1/4 compared with that of the control group, 24 h later it began to rise up slowly and then returned to the normal level comparable to the control at 120 h.

Animals↗

PubMed2006

Protein chips for high-throughput doping screening in athletes.

Sport and doping are a contradiction in terms, however, doping abuse in sports has been a serious problem for many years. The systematic screening of every athlete for all prohibited drugs should be an indispensable feature of the Olympic Games. The gas chromatography mass spectrometry method is reserved as a reference method, but is limited by its low throughput. The advent of protein chip technology may enable the screening of all athletes for any illegal use of drugs.

Doping in Sports↗

PubMed2005

Parallel detection and quantification using nine immunoassays in a protein microarray for drug from serum samples.

A protein microarray system for detection and quantification of nine prohibited drugs in serum is described. Chemically modified slides were chosen as the microarray substrates because of their suitable for drug-BSA printing. The developed protein microarray was able to preserve the biological function of the haptens, when immobilized on the microarray surface and demonstrated binding with their corresponding antibodies. The microarray could also be used for quantitative analysis when mouse IgG was chosen as an internal control for data processing. There was no qualitative difference between the results obtained using the protein microarray and ELISA. The protein microarray technology should be applicable to performing, simultaneously, large scale screening tests for many different analytes in serum.

Blood Chemical Analysis↗

PubMed2005

Primary immune effects of eukaryotic expression plasmids encoding two hyperactive mutants of human soluble B lymphocyte stimulator.

B lymphocyte stimulator (BLyS), a ligand belonging to the tumor necrosis factor (TNF) family, plays a critical role in regulating survival and activation of peripheral B cell populations during humeral immune responses. Among the TNF family members, BLyS is unique in that it contains an unpaired Cys residue (Cys146) at the corresponding position where some other members have about 37.5% (6/16) Ala or 37.5% (6/16) Val. Here, with eukaryotic expression vector pcDNA3.1(-), we mutated Cys146 to Ala or Val and constructed two mutant eukaryotic expression plasmids of the human soluble BLyS, pcDNA3.1BY-A and pcDNA3.1BY-V. Following repetitive subcutaneous injection of these expression plasmids in BALb/C mice, the wild-type and mutant BLyS proteins were detectable in the blood of treated animals over several weeks. In addition, the expression of these proteins induced specific IgG but not IgM responses. The implications of the results are discussed.

Animals↗

PubMed2004

Development of miniaturized competitive immunoassays on a protein chip as a screening tool for drugs.

BACKGROUND: Doping in sports has become a serious problem. Gas chromatography-mass spectrometry (GC-MS) serves as an effective reference method, but it is limited by low throughput and is therefore not suitable for large-scale screening. Use of protein chips for high-throughput screening of all athletes for prohibited substances could become an important complementary tool to GC-MS. METHODS: We developed a protein chip based on an aldehyde-activated glass slide containing 10 physically isolated arrays. The chip was used to screen urine from 1347 athletes for prohibited substances and to screen a negative control group consisting of 200 females and 120 males. Urine samples from 66 individuals known to be abusers, provided by the China Doping Control Center (CDCC), and 129 standard prohibited substances were tested as positive controls. RESULTS: All 1347 urine samples screened by means of the protein chips were also subjected to reference analysis by GC-MS at the CDCC. There was no qualitative difference between the results obtained with the two methods. The correlation coefficient (r(2)) for the quantitative results obtained with the protein chip and GC-MS was 0.991. CONCLUSIONS: The protein chip could be used to screen for a series of 16 prohibited drugs in urine samples. This system has the potential to become an effective screening method to test substances prohibited by the International Olympic Committee.

Doping in Sports↗

PubMed2004

Preparation of steroid antibodies and parallel detection of multianabolic steroid abuse with conjugated hapten microarray.

A conjugated hapten microarray based on miniature immunoassay for fast and multiplex detection of anabolic steroids is reported for the first time. This preliminary study investigated the possibility of using a microarray technology as a multisteroid detection assay. The microarray system used eight monoclonal antibodies raised against three steroid conjugates, 4-androsten-4-chloro-17beta-ol-3-one, 1,5alpha-androsten-1beta-methyl-17beta-ol-3-one, and 5beta-androsten-1-en-17beta-ol-3-one, which were conjugated to BSA by the active ester method. In addition to 4 commercial conjugated haptens, 18 steroid-BSA conjugates were synthesized and from all these a conjugated hapten microarray was fabricated. The analyzed substances included 42 types of anabolic steroid reference materials and 28 positive urine samples. Of these, 24 anabolic steroids and 12 positive urines were successfully detected.

Anabolic Agents↗