To investigate the apoptosis-induction effect of brucine on human chronic myeloid leukemia cell line K562 cells, K562 cells were exposed to various dosages of brucine. MTT method was used to assayed the growth inhibition effect of brucine on K562 cells. The apoptosis of K562 cells was detected by acridine orange/ethidium bromide (AO/EB) double staining, Annexin-V/PI double labeling method and DNA agarose gel electrophoresis. The results showed that brucine could remarkably inhibit the K562 cell growth in a concentration-dependent and time-dependent manners at the range of 50 to 400 µg/ml, and its most significant inhibition was observed at 400 µg/ml for 72 hours and the inhibition rate was 94.0%. Staining of cells with AO-EB revealed that brucine induced nuclear chromatin condensation. After the K562 cells were treated with the brucine of 400 µg/ml for 72 hours, the most of the nucleus were orange stained and condensation-like or bead-like showing apoptotic morphology. The K562 cells treated with brucine of different concentrations (50, 100, 200, 400, 800 µg/ml) for 72 hours, Annexin-V/PI detection showed brucine could induce apoptosis of K562 cells, and apoptosis rate increased gradually with increasing concentration of drugs. The K562 cells treated with brucine of 400 µg/ml for 72 hours displayed typical ladder strap in DNA gel electrophoresis. It is concluded that brucine can efficiently inhibit cell growth and induce apoptosis of K562 cells with dose-dependent manner in concentrations of 50 - 400 µg/ml.
Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Humans ; K562 Cells ; Strychnine ; analogs & derivatives ; pharmacology

The donkey leukocyte-attenuated vaccine of equine infectious anemia virus (EIAV) was the first lentiviral vaccine that induced solid protection from the infection of virulent strains. To elucidate the mechanism of increased immunogenicity and attenuated virulence of the vaccine, the proviral genomic DNA of an EIAV vaccine strain, EIAV(DLV121) was analyzed and compared with the genome of a parental virulent strain EIAV(DV117). Full length viral genomic DNAs were amplified as two segments by LA-PCR and were cloned. Because of the genomic diversity of retroviral quasispecies, 10 full-length sequences of EIAV(DLV121) and 4 full-length sequences of EIAV(DV117) from randomly picked clones were analyzed. Results showed that the average length of the complete nucleotide sequence of EIAV(DLV121) was 8,236bp and EIAV(DV117) was 8,249bp, with the inter-strain diversity of 2.8%. As for individual genes between the vaccine and virulent strains, the differences in nucleotide sequence of S2, LTR and env were significantly higher than the other genes with the diversity of 4.1%, 3.7% and 3.1%, respectively. Considerable variations in deduced amino acid sequences were found in S2, S3 and env. The diversities were 10.4%, 5.6% and 4.8%, respectively. Furthermore, the LTR of EIAV(DLV121) consisted of at least 5 subtypes grouped by their nucleotide sequences. There were two additional N-linked glycosylation sites in the deduced amino acid sequence of EIAV(DV117) gp90 than that of EIAV(DLV121). Among glycosylation sites in the gp90 of virulent strain, 3 were found unique only in EIAV(DV117), of which 2 were located in the principle neutralizing domain (PND). In addition, there was one EIAV(DLV121) -specific glycosylation site, which was positioned in the PND, too. Taken together, it is clear that greatly increased genomic diversity exists in the EIAV vaccine strain, which provides important information for the further study on biological characters of the Chinese EIAV attenuated vaccine.
Amino Acid Sequence ; Animals ; Base Sequence ; Equidae ; Genome, Viral ; Infectious Anemia Virus, Equine ; chemistry ; genetics ; immunology ; Leukocytes ; immunology ; virology ; Molecular Sequence Data ; Sequence Alignment ; Vaccines, Attenuated ; chemistry ; genetics ; immunology ; Viral Proteins ; chemistry ; genetics ; immunology ; Virulence

OBJECTIVETo investigate the relationship between acute graft-versus-host disease and graft composition in patients with aplastic anemia(AA) after haploidentical hematopoietic stem cell transplantation.

METHODSFifty-seven cases of AA after haploidentical hematopoietic stem cell transplantation were retrospectively analyzed. All the patients were divided into 2 groups according to whether presence or absence grade Ⅱ-Ⅳ aGVHD, the relationship between aGVHD and graft composition was analyzed by comparing the differences of graft components between the 2 groups.

RESULTSFourteen out of 57 patients had grade Ⅱ-Ⅳ aGVHD and the other 43 did not have grade Ⅱ-Ⅳ aGVHD. The mononuclear cells, CD3, CD4, CD8, NK cells, NKT cells, B cells and Treg cells were not significantly different between the 2 groups (P>0.05), the CD34 cell count in the patients with grade Ⅱ-Ⅳ aGVHD was 3.85（1.73-10.61）×10/kg, which was significantly lower than that without grade Ⅱ-ⅣaGVHD: 6.31（2.98-19.35）×10/kg (P<0.05).

CONCLUSIONSThe incidence of grade Ⅱ-Ⅳ aGVHD may be related with CD34 cell count in AA after haploidentical hematopoietic stem cell transplantation..