AIM:To determine the effects of Tongxinluo (TXL) on connexin 43 (Cx43) remodeling and ven-tricular arrhythmia ( VA) after myocardial infarction ( MI) in rats .METHODS:Male SD rats were randomly divided into sham-operated (sham) group (n=25) and operation group (n=75).The left anterior descending (LAD) was ligated in operated group , while the rats in sham group only underwent pericardiotomy .The rats in operation group which survived for 3 d after operation were randomly assigned to TXL group and MI group .The rats in TXL group was administrated with TXL (2 g? kg-1? d-1, intragastric administration) for 4 weeks, while normal saline was applied to the rats in sham group and MI group.The levels of interleukin-1β(IL-1β) and endothelin-1 (ET-1) in the tissue from the border zone were measured by ELISA after treatment .The distribution and the mRNA and protein expression of Cx 43 were detected by immunohisto-chemical staining , RT-PCR and Western blotting , respectively .The burst pacing was used to induce ventricular arrhythmia ( VA) .RESULTS:Compared with sham group , the levels of IL-1βand ET-1 and the incidence of VA were significantly increased , while the mRNA and protein expression of Cx 43 was markedly reduced with irregular distribution in MI group (P<0.05).Compared with MI group, the levels of IL-1βand ET-1 and the incidence of VA were significantly reduced , while the expression of Cx 43 at mRNA and protein levels was markedly increased with augmented linear distribution in the myocardial cell intercalated disc in TXL group (P<0.05).CONCLUSION:TXL reduces the incidence of VA after MI via inhibiting the Cx43 remodeling .

To screen the specific anti-human intercellular adhesion molecule-1 (ICAM-1) single chain fragment variable (scFv) using phage display library technology and to identify its biological activity. P1 peptide was used as antigen, and the phage antibodies against human ICAM-1 antigen were panned by four binding-eluting-amplifying cycles using Tomlinson I+J phage display library. After four rounds of selective enrichment screening, the positive clones were determined by PCR, enzyme linked immunosorbent assay (ELISA)-based antigenic cross reaction and Dot blotting. Then the binding specificity and biological activity of purified scFv were identified by Western blotting, competitive ELISA and cell adhesion inhibition assay respectively. Furthermore, four positive clones were first panned through P1 peptide coated-ELISA assay, and then J-A1 was obtained and identified by PCR, ELISA-based antigenic cross reaction and Dot blotting, which could show a specific binding between P1 peptide and human ICAM-1 protein antigen. Subsequently, the purified scFv showed a satisfactory specificity and anti-adhesive activity in competitive ELISA and the cell adhesion inhibition assay. The specific anti-human ICAM-1 scFv was prepared successfully from Tomlinson I+J phage display library, which pave the way for further application of anti-human ICAM-1 scFv for inflammation diseases therapeutics.
Antibodies ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunoglobulin Variable Region ; Intercellular Adhesion Molecule-1 ; immunology ; Peptide Library ; Single-Chain Antibodies