ABSTRACT
AIM: To choose the anti myocardial ischemic ingredients from polysaccharide of Ophiopogon Japonicus(PO), then to confirm their effects. METHODS: According to molecular weight, PO was separated to three portions by using ultrafiltration technique. The effects of each separated portion on myocardial ischemia induced by isoproterenol were studied. Then the effects of PO active portions (POAP) on the convulsion of coronary artery induced by pituitrin and acute myocardial ischemia caused by the ligation of coronary artery were further studied in rats. RESULTS: POAP could resist the elevation of Tand S T phase induced by pituitrin in rats, remarkably decrease the elevated S T phase and contents of CK and LDH in serum caused by the ligation of rat coronary artery. POAP also exerted some inhibition effects on the reduction of SOD and increase of MDA caused by myocardial ischemia. CONCLUSION: POAP can protect myocardial cells, inhibit the generation of free radicals, and scavenge the oxygen free radicals generated from myocardial ischemia.
ABSTRACT
190-kilodalton glycoprotein (P190) of Plasmodium falciparum. precursor of the major surface protein of merozoites, is considered a promising candidate for blood stage malarial vaccine. We designed six primers according to the sequence of MAD20 strain, with a GC clamp and BamHI site at the 5'- end of each one, and a GC clamp and Xbal site at the 3'- end of each one. The primers were synthesized by phosphoramidite approach (User's Manual of ABI Company) and purified using HPLC. Three fragments in the second, third and fourth conserved regions of P190 gene of Plasmodium falciparum FCC1/HN strain isolated from the blood of patients in Hainan Province of China were amplified by the polymerase chain reaction (PCR) technique. The amplified fragments were subcloned into pUC18 vectors and sequenced using the dideoxy chain termination method. All three regions of P190 gene of FCCl/HN strain also were highly conservative as compared with P190 gene of MAD20 (Papua New Guinea isolate), K1 (Thailand isolate), Wellcome (West Africa isolate) and CAMP (Malaysia) strains of Plasmodium falciparum. The C at position 81 in the second conserved block of P190 gene of FCC1/HN isolate was substituted by T, which did not change the amino acid determined by the coden corresponding to the substitution.
ABSTRACT
Two DNA fragments, designated as P190CRI (AA1-55) and P190CRV (AA1597-1667) respectively, which encoded amino acid residues of conserved region I and V on the P190 antigen, were amplified by polymerase chain reaction from genomic DNA in FCC1/HN strain of Plasmodium falcipamm isolated from Hainan Province, China. It was found that there were five bases substitution in the P190CRV, in comparison with the nucleotide sequences of MAD20 strain. These two sequenced fragments were inserted into pGEX-2T plasmid. E.coli JM109 (DE3) were transformed with the recombinant plasmids and the parental plasmid. The results show that the two fragments were expressed at high level as C-terminal fusions with glutathione s-transferase (GST). The fusion proteins were easily purified from bacterial lysates by affinity chromatography using glutathione sepharose 4B.