Gengqian Zhang

Construction and Preliminary Investigation of a Plasmid Containing a Novel Immunotoxin DT390-IL-18 Gene for the Prevention of Murine Experimental Autoimmune Encephalomyelitis

Experimental autoimmune encephalomyelitis (EAE) is a neuropathological animal model for multiple sclerosis. Antigen-presenting cells (APCs) expressing interleukin-18 receptor (IL-18R) were shown to be crucial in the beginning and progress of EAE. In this study we tested the effect of a novel recombinant immunotoxin targeting IL-18R-bearing APC for EAE prevention. The novel eukaryotic plasmid DT390-IL-18-SRalpha, encoding recombinant immunotoxin DT390-IL-18, was constructed. The immunotoxin consisted of IL-18 as the targeting moiety, and a truncated diphtheria toxin (DT) as the toxic moiety. Transfection assay and proliferation inhibition assay proved the immunotoxin could be expressed in vitro and was toxic to the activated mouse T cells. To evaluate the preventive effect of DT390-IL-18-SRalpha on EAE in vivo, cationic liposome-embedded DT390-IL-18-SRalpha was injected into the hind limbs of EAE mice. DT390-IL-18-SRalpha-treated mice showed a delayed manifestation of EAE and decreased symptoms compared to the mice treated with plasmid DT390-SRalpha or phosphate-buffered saline alone. A significant reduction in infiltrating inflammatory cells was detected in the brain tissues from immunotoxin-treated mice as compared with the controls by hematoxylin-eosin staining. This study suggested that the recombinant immunotoxin DT390-IL-18 could be expressed in vitro and in vivo, and prevented murine EAE effectively.

Time course proteomic profile of rat acute myocardial infarction by SELDI-TOF MS analysis

BACKGROUND Surface-enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF MS) is a powerful tool to detect biomarkers of many diseases. The purpose of this study is to evaluate SELDI-TOF MS as a potential tool for identifying serological biomarkers of acute myocardial infarction (AMI) at an early stage. METHODS Serum samples were collected from rats after they had received left anterior descending coronary artery (LAD) ligation at 5, 15, 30, 60, 120, 240 and 360 min, respectively. These samples, along with serum samples from the sham-operated and uninjured control animals, were processed and analyzed with CM-10 ProteinChip and SELDI-TOF MS. Levels of myocardial creatine kinase isoenzyme MB (CKMB) activity and cardiac troponin I (cTnI) concentration were measured with respective commercial kits. Histological changes were examined after HE staining. RESULTS Three peaks with m/z 7586 Da, 7564 Da and 9583 Da, which were only found in the operated groups, were termed specific peaks. The peaks with m/z 7586 and 7564 Da had the sensitivity of 97.0% and specificity of 97.0% in the >or=15 min operated groups. Four peaks with m/z 4983, 5140, 8075, and 9423 Da were novel tissue unspecific injury related peaks in response to injury. Four peaks with m/z 4400, 4542, 8400 and 7666 Da exhibited more than 2 fold peak height alterations. In contrast, the significant increase of CKMB activity and cTnI concentration in the serum could not be detected until 4 h after the ligation. CONCLUSIONS The specific peaks and unspecific injury related peaks detected by SELDI-TOF MS with CM-10 ProteinChip may be used as the biomarkers for early diagnosis of AMI.

Allele frequencies for two STR loci D6S1274 and D17S1299 in Chinese and Thai populations.

POPULATION: Chinese Han, Thai.