Objective To explore the possibility of cytochrome P-450 3A5 (CYP3A5) genotype as a major factor to guide individualized employment of cyclosporin A(CsA) through a comparative study of CsA concentrations in the peripheral blood of hemopoietic stem cell transplant recipients with different CYP3A5 genotypes. Methods Olymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to genotype CYP3A5 gene, and CsA concentrations were detected by a commercial fluorescence polarization immunoassay. Result There were significant differences in CsA concentrations, including standardized trough concentrations C0 and two h peak concentrations C2 , in the two CYP3A5 genotypes found in our samples, and both C0 and C2 in wild homozygotes were lower than heterozygotes. Conclusion CYP3A5 polymorphism is highly associated with CsA concentrations in hemopoietic stem cell transplant recipients, and CYP3A5 genotype may be a predictor to the dose requirement before clinically employment of CsA.

Objective To discriminate morphology and immunophenotype differences between hematogones and lymphoblast to provide some references for the correct identification of hematogones and minimal residual leukemia cells.Methods Immunophenotypes were detected by flow cytometry in a total of 132 bone marrow from 58 patients with acute B lymphoblastic leukemia during diagnosis,remission and relapse.Hematogones were identified based on combination of CD34/CD10/CD19/CD45 or CD34/CD10/CD45/CD19/CD20/CD38.Results Among 132 specimens,45 (34 ％) were identified hematogones,the detection range was 0-36 ％.Three specimens appeared in diagnosis patients,one in relapse,and the remaining 41 cases in remission.The detection rate of hematogones was 62 ％ (41/66) in the remission cases.More than 5 ％ leukemia cells of nucleated cells were detected in diagnosis and relapse,and less than 5 ％ residual leukemia cells was in 24 specimens from remission patients.In 28 specimens,the co-existence of hematogones and leukemia cells was found,including three in diagnosis,one in relapse and the remaining 24 in remission.Hematogones were characterized in term of variable expression of CD45 and very low side scatter.The early hematogones expressed CD34.With maturation increasing,hematogones gradually lacked CD34.CD19 and CD10 were presented in whole hematogones stage.Early hematogones had expression of CD10.Lymphoblasts showed maturation arrest and more homogeneous populations.SSC values of hematogones were higher than that of normal B cell progenitors.Antigen overexpression or underexpression was not found in normal hematopoietic progenitor B cells,and hematopoietic progenitor B cells did not appear cross-lineage markers,CD20+ cells exhibited continuous distribution from negative to weak positive for normal hematogones.Conclusions Hematogones were present in diagnosis,remission and relapse cases with acute B lymphoblastic leukemia,especially abundant in bone marrow after chemotherapy.It should be careful to diagnose and discriminate the malignant cells from benign cells.By comprehending continuous and complete maturation spectrum of antigen expression for normal hematogones,knowing phenotype of leukemia cells drift change patterns and using multiparameter flow cytometry and optimal antibody combination,it is significant in identifying residual lymphoblasts from hematogones and improving the detection accuracy in minimal residual disease.

Objective To construct a new molecular biological method for the analysis of microbial species in lower respiratory tract infections based on 16S rRNA gene by denaturing high-performance liquid chromatograph(DHPLC).Methods The universal primer set was analyzed basing on the highly conserved regions of 16S rRNA gene.DNA amplicons of lower respiratory tract were analyzed by DHPLC to generate peak profiles respectively.The incorporation of 40-bpGC clamp into the amplification primet was essential to effectively discriminate genetic differences identification.Results The primers could only amplify bacterial 16S rRNA.Bacterial of amplicons which incorporation of a 40-bpGC clamp were effectively discriminated genetic differences in DHPLC.The results of clinical isolares identification showed 100%according with the traditional method.Conclusion DHPLC has not only high accuracy,but also is a convenient,rapid and high-through technique for the discrimination bacteria.It has potential value in the detection of lower respiratory pathogenic bacteria.

Objective To investigate the significance of fluorescence in situ hybridization (FISH) panel in detecting cytogenetic abnormalities in patients with chronic lymphocytic leukemia (CLL).Methods A panel of FISH probes [D13S25 (13q14.3),RB1 (13q14),ATM(1 1q22.3),CSP12(12p1 1.1-12q1 1.1) and p53 (17p13.1)] were performed in 21 cases with CLL.The cytogenetic features in correlation with clinical manifestation,other laboratory tests and prognosis were analyzed.Results Cytogenetic abnormalities were found in 13 of 21 patients with CLL (61.90 ％).The most frequent abnormality was del(13q14) (42.86 ％),followed by trisomy 12 (14.29 ％),del(11q22) (9.52 ％) and del(17p13) (9.52 ％).There was no significant relationship among cytogenetic abnormalities and sex,binet stages,expression of CD38,level of lactate dehydrogenase.Conclusion FISH with probe panel is a rapid,sensitive and accurate technique for detection of cytogenetic abnormalities in patients with CLL.

Objective To investigate the target genes in RYBP-mediated leukemia cell apoptosis by high-flux sequencing. Methods The HL-60 cell line with knockdown of RYBP was set up. MRNA and microRNA sequencing was conducted by the next-generation sequencing technology. Result MRNAs and miRNAs were dys-regulated in HL-60 cell line with knockdown of RYBP. The results of KEGG analysis indicated that the down-regu-lated genes were associated with cancer transcription regulation and amino acid metabolism. SPI1 and miR-214-3p were shown downregulated after knockdown of RYBP. Conclusion The target genes in RYBP-mediated leukemia cell apoptosis include BBC3,BAI1,SESN2 ,CCNG2,JAK3,STAT4,SPI1,BCL6,CD11b and hsa-miR-214-3p.