Expression and clonal proliferation of TCR Vbeta subfamilies of peripheral T-cells in acute myeloid leukemia patients / 中国实验血液学杂志

This study was purposed to investigate the expression and clonal proliferation of receptor (TCR) Vbeta subfamilies of the T-cells in acute leukemic patients at different disease status (onset, complete remission or relapse) and to analyze the influence of the leukemic cell load on anti-leukemic effect of peripheral T-lymphocytes of the patients. Gene sequences of peripheral TCR Vbeta 24 families from 11 leukemic patients and 3 normal donors were expanded by RT-PCR. Genescan technique was applied to evaluate clonal expression of the TCRVbeta subfamilies, clonal characteristics of the CDR3 from peripheral blood of AML patients at different disease status. The application, clonal proliferation, cellular complexity of T-cells, and the variation of immunotypes of T-cells were compared. The results indicated that the lower and partial distribution of TCR Vbeta subfamily was found in all 11 patients when firstly diagnosed; the expression of TCR Vbeta subfamilies after induction in vitro increased; obvious elevation of TCR Vbeta subfamilies was observed in patients at complete remission although expression level was still lower than normal, whereas the significant descent of TCR Vbeta subfamilies was detected in 4 relapsed patients. Only 1 - 2 clonal proliferation of TCR Vbeta subfamilies existed in 9 out of 11 patients at initial diagnosis which increased at remission. The status of clonal proliferation of Vbeta subfamily T-cells continued regardless of any different disease status in most patients. There was an obvious decrease of CDR3 complexity at initial diagnosis or relapse, while CDR3 complexity would be partially improved at remission. It is concluded that the restrict distribution and expression of TCR Vbeta subfamilies were found in AML patients. Clonal proliferation of T-cells Vbeta subfamily continuously exists regardless of any different disease status in most patients. Some Vbeta subfamilies sustain clonal proliferation at different disease status. Some clonal proliferations of Vbeta subfamilies are associated with the effects of leukemic cells, CDR3 complexity obviously decreases under disease status which can be partially improved at remission.

Study on polymorphisms of CYP3A5 gene and their clinical role / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the polymorphisms of CYP3A5 gene in Chinese population and the association between CYP3A5 genotypes and their clinical functions.</p><p><b>METHODS</b>CYP3A5 gene varisances were detected in 180 samples using denaturing high-performance liquid chromatography(DHPLC), and CsA concentrations in 12 of 180 samples from hemopoietic stem cell transplant recipients were monitored by a commercial fluorescence polarization immunoassay. The data were analyzed by a statistical software.</p><p><b>RESULTS</b>In the 180 samples, there was only one allelic variant CYP3A5*3 with a frequency of 76.1% (274/360), and there were three CYP3A5 genotypes, namely CYP3A5*1/*1, CYP3A5*1/*3 and CYP3A5*3/*3 with frequencies of 5.6%, 36.7% and 57.8% respectively. Also, there were significant differences in CsA concentrations, including standardized trough concentrations C(0) and two-hour peak concentrations C(2), between CYP3A5 CYP3A5*1/*1 and CYP3A5*1/*3 found in 12 hemopoietic stem cell transplant recipients, and both C(0) and C(2) in CYP3A5*1/*1 were lower than those in CYP3A5*1/*3.</p><p><b>CONCLUSION</b>CYP3A5*3 is the primary allelic variant in Chinese population. CYP3A5 genotypes are closely associated with blood CsA concentrations in hemopoietic stem cell transplant recipients, and CYP3A5*1/*1 requires a larger CsA dose to maintain the same blood concentration than does CYP3A5*1/*1. CYP3A5 genotyping by DHPLC may predict recipients' phenotype and CsA dose requirement.</p>

A novel mutation in DKC1 gene in a patient with dyskeratosis congenita / 中华皮肤科杂志

Objective To detect the mutation in DKC1 gene in a patient with dyskeratosis congeni- ta.Methods Fifteen exons of DKC1 gene were amplified by polymerase chain reaction (PCR),and the products were screened for mutations by denaturing high performance liquid chromatography (DHPLC) technology,then DNA sequencing was performed for abnormal exons as shown by DHPLC.The gene muta- tions were verified within 100 unrelated male individuals without dyskeratosis congenita.Results An ab- normal DHPLC elution peak was found in exon 12 of DKC1 gene of the patient,but not in other family members or normal individuals.DNA sequencing showed a 1236G→T transition in DKC1 gene in the pa- tient,which resulted in a 412W→C substitution in DKC1.No mutation was found in other family members and normal individuals.Conclusion The 1236G→T transition in the patient is a novel mutation in DKC1 gene,which could be a causative factor of dyskeratosis congenita.