[Hematopoietic reconstitution after transplantation of uncontrolled-rate cryopreservation autologous peripheral blood hematopoietic stem cells using -80 °C mechanical freezer].

Objective: This study was to identify the efficacy of -80°C cryopreservated peripheral blood hemato-poietic stem cell (PBHSC) transplantation for hematopoietic reanstitution in patients.

Methods: The efficacy of 104 patients underwent autologous peripheral blood hematopoietic stem cell transplantation using uncontrolled-rate freezing and storage at -80°C was evaluated.

Results: This cryopreservation method could effectively cryopreserve peripheral blood stem cells.

[HLA antigen compatibility between patients with hematologic diseases and their parents].

This study was aimed to investigate and analyze the HLA antigen compatibility between patients with hematologic diseases and their parents so as to provide basis for selecting the suitable donors in haploidentical hematopoietic stem cell transplantation. The HLA low resolution for 174 families was typed and analyzed by using PCR-SSP. The results showed that 52.

[Clinical analysis for 3 cases of HLA-matched between father and son and 1 case of post-hematopoietic stem cell transplant efficacy].

Getting a HLA-matched donor is a key factor for successful hematopoietic stem cell transplantation. People are almost semi-matched with their parents, while a person HLA-matched with his/her father or mother was rarely seen, if so, usually whose father and mother are genetically related. HLA-low resolution for patients and their relatives were performed using PCR-SSP technique and three patients were found HLA-matched with their father in these results.

[Application of spectral karyotyping to cytogenetic analysis in acute myeloid leukemia].

Objective: To evaluate the value of spectral karyotyping (SKY) in cytogenetic analysis of acute myeloid leukemias (AML).

Methods: Nine AML patients were analyzed by R-banding and SKY. MLL, PML-RARalpha, AML1-ETO fusion genes were detected by dual fusion- fluorescence in situ hybridization (D-FISH).

[A recombination event occurring between HLA-A and -A loci from father's HLA haplotype chromosome].

This study was aimed investigate the recombination event occurring between HLA-A and-A loci discovered from father's HLA haplotype chromosome in a family. Peripheral blood samples were collected from a family. HLA class I (-A, -B, and -Cw) and II (-DRB1 and -DQB1) alleles were amplified and typed by both low and high resolution PCR with sequence-specific primers (PCR-SSP) and sequence-based typing (SBT).

[Allogeneic hematopoietic stem cell transplantation combined with imatinib for treatment of Philadelphia chromosome positive acute lymphoblastic leukemia].

The study was purposed to explore the efficacy of allogeneic hematopoietic stem cell transplantation (allo-HSCT) combined with Imatinib for treatment of Philadelphia chromosome positive acute lymphoblastic leukemia (Ph(+)ALL) patients. From 2007 to 2008, 3 patients with Ph(+)ALL were treated with allogeneic hematopoietic stem cell transplantation and Imatinib, and the follow-up ended at Oct 21(st) 2009. 1 patient received HSCT from matched sibling donor and 2 patients from haploidentical related donors.

[Killer immunoglobulin-like receptor gene genotype and expression pattern in NK-92MI and K562 cells].

Aim: To make clear the repertoire of Killer immunoglobulin-like receptor at the level of DNA and RNA in NK-92MI and K562 cells and compare KIRs gene genotype and expression patterns in both cells.

Methods: KIRs gene genotype, include KIR2DL1, 2DL2, 2DL3, 2DL4, 2DL5, 2DS1, 2DS2, 2DS3, 2DS4, 2DS5, 3DL1, 3DL2, 3DL3, 3DS1, 2DP1, 3DP1, was analyzed by PCR. KIRs gene phenotype, include KIR2DL1, KIR2DL2, KIR2DL3, KIR3DL1, KIR2DS1, KIR2DS3, KIR2DS2/4 gene, was determined by RT-PCR and sequencing.

[Effect of demethylation treatment on the expression of inhibitory receptor KIR gene in NK-92MI cell line].

The aim of this study was to analyze the promoter methylation patterns of inhibitory killer cell immunoglobulin-like receptor (KIR) which gene expression and the effect of demethylation treatment were studied, and to explore the possible regulation mechanism of inhibitory kir gene expression. The promoter methylation levels of kir2DL1 and kir2DL2/kir2DL3 in NK-92MI cell line were detected by bisulfite sequencing technique. Then NK-92MI cells were treated with 5-azacytidine to induce the demethylation of CpG islands.

[Investigation on MS-PCR as a method for detecting minimal residual disease in acute leukemia].

The aim of this study was to investigate the feasibility of monitoring minimal residual disease (MRD) of leukemia with methylation specified-polymerase chain reaction (MS-PCR). The HL-60 cells with Id4 gene complete methylation and Hek937 cells with Id4 gene complete unmethylation were mixed in accordance with different ratios of cells and were divided into 3 groups: group A (10% HL-60 + 90% Hek937), group B (1% HL-60 + 99% Hek937) and group C (0.1% HL-60 + 99.

[Dynamic detection of chimerism and fusion gene in chronic myeloid leukemia patients relapsed after allogeneic hematopoietic stem cell transplantation].

This study was aimed to investigate the chimerism and fusion gene expression in patients with CML after allo-HSCT, to analyse engraftment and minimal residual disease by using STR-PCR combined with RT-PCR qualitative and quantitative assays, and to evaluate their clinical value for predicting disease relapse. 4 relapsed patients with CML after allo-HSCT were dynamically investigated. Qualitative analysis of donor chimerism was performed by multiplex PCR amplification of STR markers and capillary electrophoresis with fluorescence detection, qualitative detection of bcr/abl transcripts was performed by RT-PCR.

[Construction and identification of Kir2ds4 RNAi lentiviral vector].

This study was aimed to construct a lentiviral vector of RNA interfered (RNAi)-kir2ds4 gene. In accordance with study-confirmed effective sequence of siRNA targeting kir2ds4 gene, the complementary DNA containing both sense and antisense oligonuctide of the targeting sequence was designed, synthesized and inserted into pSicoR-GFP vector containing U6 promoter and GFP sequence. The resulting lentiviral vector containing kir2ds4 shRNA was named as LV-sh kir2ds4, and confirmed by PCR and sequencing.

The diversity of KIR gene in Chinese Northern Han population and the impact of donor KIR and patient HLA genotypes on outcome following HLA-identical sibling allogeneic hematopoietic stem cell transplantation for hematological malignancy in Chinese people.

Killer cell immunoglobulin-like receptors (KIRs) are members of a group of molecules that specifically recognize HLA class I ligands and are found on subsets of human lymphopoetic cells. The number of KIR loci can vary between individuals, resulting in a heterogeneous array of possible KIR genes. The range of observed profiles has been explained by the occurrence of two haplotype families termed A and B, which can be distinguished on the basis of certain KIR sequences.

[Methylation status of ZO-1 gene in patients with myelodysplastic syndrome].

The objective of this study was to investigate the methylation status of zonula occluden protein-1 (ZO-1) gene in patients with myelodysplastic syndrome (MDS) and to identify its roles in pathogenesis, development and classification of MDS. 85 patients with MDS and 30 healthy individuals were tested by methylation specific polymerase chain reaction (MS-PCR). The results indicated that no ZO-1 promoter methylation could be detected in healthy controls.

[Application of chimerism analysis to allogeneic hematopoietic stem cell transplantation by STR-PCR].

The aim of this study was to analyze chimerism, evaluate the status of engraftment and predict the outcome of allogeneic hematopoietic stem cell transplantation (allo-HSCT) by multiple short tandem repeat (STR) amplification using fluorescence labeling polymerase chain reaction (PCR) combined with capillary electrophoresis. Peripheral blood and bone marrow in 27 patients who received myeloablative allogenetic cell transplantation were collected before and after transplantation in different times. 10 and 7 different STR markers were co-amplified in a single reaction by using a commercial AmpF/STR Profiler Plus/Cofiler plus PCR amplification kits.

[Primary application of spectral karyotyping in leukemia].

Objective: To establish a spectral karyotyping (SKY) technique and explore the value of SKY in leukemia research.

Methods: SKY technique was conducted on 2 samples of peripheral blood from 2 healthy volunteers, then on the samples from 8 patients with leukemia, including chronic myelocytic leukemia (CML) and acute myelocytic leukemia (AML) confirmed by R-banding. In addition, four patients underwent dual fusion-fluorescence in situ hybridization (DF-FISH) to detect the mixed lineage leukemia (MLL), PML/RARa, and BCR/ABL fusion genes.

[Effect of compound recipe Gengniankang on senile sexual hormone and expression of estrogen receptor in bone of climacteric female rats].

Objective: To compare the therapeutic effect of Compound Recipe Gengniankang ( GNK) with that of hormone replacement treatment (HRT) on climacteric female rats with osteoporosis, and to investigate the roles of estrogen and estrogen receptors in the mechanism of osteoporosis.

Methods: Climacteric female rats with osteoporosis were chosen and divided into three groups (GNK group, HRT group and control group). Apoptosis of ovarian granulose cells was measured by terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) assay.

[Experimental study on regulatory effect of compound gengniankang on endocrine and immune functions in aged female rats with osteoporosis].

Objective: To observe the effect of compound gengniankang (GNK) in regulating the endocrine and immune functions in aged female rats.

Methods: Aged female rats with osteoporosis were selected as the object for observation and healthy young rats were taken for control. Animals were administered by GNK and nilestriol respectively, through gastric perfusion, for 3 months to observe the therapeutic effect of drug treatment on osteoporosis and the regulatory effect on endocrine and immune function.