[Effects of Leukapheresis on Hemostatic Function in Patients with Hyperleukocytic Leukemia].

Objective: To analyze and compare the effects of leukapheresis on hemostatic function in patients with hyperleukocytic leukemia.

Methods: A total of 139 patients with AML, ALL and CML who underwent leukapheresis from June 2009 to February 2020 and did coagulation test before and after operation were included in this study. The clearance efficiency of each group and the difference among three groups were evaluated, as well as hemostatic function including platelet counts, coagulation indicators, CDSS score and incidence of adverse events.

[Effects of Simvastatin on Expression of Multidrug Resistance Gene and Protein of SHI-1 Cells].

Objective: To investigate the effects of simvastatin(SIM) and serum free medium(SFM) on the expression of multidrug resistance gene(MDR1) and protein of SHI-1 cells.

Methods: Trypan blue exclusion assay was used to detect the proliferation level and viability of SHI-1 cells after treatment with SIM and culture in SFM; The multi-drug resistant protein p-gp was measured by flow cytometry after culture in SFM for 1 to 3 days and treatment with various concentration of simvastatin. The effect of SFM culture and SIM treatment on the expression of MDR1 trascript was detected by qPCR; ELISA was used to measure the change of cellular cholesterol after culture in SIM and SFM.

Comparison of SPE, IFE, and FLC in Monitoring Patients with Multiple Myeloma After Autologous Stem Cell Transplantation.

Conventionally, serum protein electrophoresis (SPE) and serum immunofixation electrophoresis (IFE) are used as primary methods to diagnose and monitor multiple myeloma (MM). Recently, serum-free light chain (FLC) assay has been incorporated into hematological screening programs for myeloma. The purpose of this study is to compare the performance of the three methods in monitoring MM patients after autologous stem cell transplantation (ASCT).

[Ex vivo inducing cultured Epstein-Barr virus specific cytotoxic T lymphocytes and evaluation of their killing effect].

This study was aimed to explore the method for induction and expansion of EB virus specific cytotoxic T lymphocytes (EBV-CTL) in vitro, and to detect their killing effect. Peripheral blood mononuclear cells (PBMNC) were collected from 6 EBV seropositive healthy donors, and EBV-transformed B lymphoblastoid cells (BLCL)were used as the antigen-presenting cells and antigen stimulant which was irradiated by 40 Gy (60)Co irradiator. The autologous PBMNC and irradiated BLCL were cultured to induce and expand the EBV-CTL, and the immunophenotype was identified by the flow cytometry.

[Quantitative monitoring of multi-donor chimerism after multi-donor allogeneic hematopoietic stem cell transplantation].

This study was aimed to establish a model for detecting the donor chimerism rate following the multi-donor hematopoietic stem cell transplantations, and simplify its calculation method. Patients with hematologic disease receiving allogeneic hematopoietic stem cell transplantation including single-donor and multi-donor were selected in this study and the donor cell chimerism rates were detected, using STR-PCR combined with capillary electrophoresis. The results indicated that the peaks of the sister alleles coming from the same individual were confirmed to have the approximate areas and can be replaced each other in the situation of mixed chimerism.

[The correlation of cytomegalovirus gB genotype with viral DNA load and treatment time in patients with CMV infection after hematopoietic stem cell transplantation].

Objective: To explore the effect of CMV gB genotypes on viral load and treatment time in patients with CMV infection after hematopoietic stem cell transplantation (HSCT).

Methods: Viral load was detected by real-time (RT) quantitative polymerase chain reaction (PCR) (Q-PCR), CMV gB genotypes by PCR restriction fragment length polymorphism (RFLP) (PCR-RFLP) in 115 patients with CMV infection (CMV-DNA positive) after HSCT during July 2004 and May 2010.

Results: (1) The distribution of CMV gB genotypes in HSCT recipients were as following: gB1, 42/115 (36.

[Culture and pluripotentiality of murine compact bone-derived mesenchymal stem cells].

This study was purposed to culture murine compact bone-derived mesenchymal stem cell (MSC) and analyze the immunological and trilineage differentiation potential. Tibia and femur were extracted. Bone marrow cells were flushed out and compact bone fragments were digested with collagenase.

[Actions of keratinocyte growth factor in leukemic mice allogeneic umbilical cord blood cell transplantation].

Objective: To explore the actions of keratinocyte growth factor (KGF) in leukemic mice allogeneic umbilical cord blood cell transplantation (UCBT) and elucidate its mechanism.

Methods: Peripheral blood drawn from the litters of C57BL/6 females was used as umbilical cord blood (UCB) graft. BALB/c mice were randomly divided into 7 groups (n = 12 each).

[Effects of simvastatin on proliferation and apoptosis of acute monocytic leukemia cell line SHI-1].

The purpose of this study was to investigate the effect of simvastatin (SIM) on proliferation and apoptosis of acute monocytic leukemia cell line SHI-1 and its mechanism. Experiments were divided into control and test groups (5 µmol/L, 10 µmol/L, 20 µmol/L SIM groups). The growth inhibitory rate of SHI-1 cells was detected using methyl thiazolyl tetrazolium (MTT) method.

[Relationship between CMV reactivation and KIR haplotype/HLA-Cw genotype in patients after unrelated-donor hematopoietic stem cell transplantation.].

Objective: To explore the relationship between CMV reactivation and KIR haplotype or HLA-Cw genotype in patients after unrelated-donor hematopoietic stem cell transplantation (HSCT).

Methods: From January 2003 to December 2008 the HLA-Cw/KIR genotype of 48 patient-donor pairs were determined by polymerase chain reaction with sequence specific primers (PCR-SSP) and sequence specific nucleotide (PCR-SSOP). Posttransplant CMV reactivation was performed by immune histochemically assay.

[Detection and clinical significance of JAK2 mutation in 412 patients with chronic myeloproliferative neoplasms].

Objective: To investigate the frequency of JAK2V617F mutation in Chinese patients with chronic myeloproliferative neoplasms (MPN) and to study the relationship between JAK2V617F mutation and clinical characteristics.

Methods: JAK2V617F mutation was screened by allele-specific polymerase chain reaction (AS-PCR).

Results: JAK2V617F mutation was detected in 277 of the 412 patients with MPN.

[A quantitative assay for JAK2 mutation in 135 patients with chronic myeloproliferative neoplasms].

Objective: To investigate the frequency and mutational status of JAK2 mutation in Chinese patients with chronic myeloproliferative neoplasms (MPN) and study the relative quantitation and clinical implications of mutated JAK2 transcript.

Methods: JAK2 mutation and the mutational status were screened with amplification-refractory mutation sequencing polymerase chain reaction (ARMS-PCR), the relative quantity of mutated JAK2 mRNA by using capillary electrophoresis.

Results: JAK2V617F mutation was detected in 95 of 135 MPN patients, including 37 (97.

[Detection of JAK2V617F mutation and its clinical significance in 80 patients with M2 acute myelogenous leukemia].

Objective: To explore the prevalence and prognostic significance of JAK2V617F gene mutation in acute myelogenous leukemia M2 (AML-M2) patients.

Methods: Allele specific polymerase chain reaction (PCR) was used to detect JAK2 gene mutation.

Results: Of 80 de novo AML-M2 patients, 6 at the time of first diagnosis and 1 at relapse were found to have JAK2V617F gene mutation (8.

[5-azacytidine treatment induces tumor suppressor gene XAF1 expression and inhibits proliferation in myeloma cells].

Objective: To investigate the effect of 5-azacytidine on XAF1 expression in myeloma cell lines RPMI8226 and XG-7 and the in vitro anti-myeloma activity of 5-azacytidine.

Methods: XAF1 mRNA and protein expression was detected by semi-quantitative reverse transcriptase PCR and Western blot, respectively. Methylation specific PCR (MSP) was used to detect methylation status of XAF1 promoter CpG islands.

[The quantitative assay and clinical significance of JAK2V617F mutation in 131 patients with chronic myeloproliferative disorders].

Objective: To investigate the frequency and mutational status of JAK2V617F mutation in Chinese patients with chronic myeloproliferative disorders (CMPD) and to study the relative quantification of mutated JAK2 mRNA and the clinical significance.

Methods: JAK2V617F mutation and the mutational status were screened with amplification-refractory mutation system polymerase chain reaction (ARMS-PCR), the relative quantification of mutated JAK2 mRNA was studied by using capillary electrophoresis.

Results: A higher prevalence of JAK2V617F in either the heterozygote or homozygote status in essential thrombocythemia (ET) was observed in elderly patients with ET (P < 0.

[Quantitative analysis for JAK2 mutation in 98 patients with essential thrombocythemia and its clinical significance].

The objective of this study was to identify the frequency and types of JAK2V617F mutation in chinese patients with essential thrombocythemia (ET), to quantitatively detect the level of mutation transcripts and to investigate its clinical significance. The frequency and types of JAK2V617F mutation were detected by amplification-refractory mutation sequencing polymerase chain reaction (ARMS-PCR), the transcript level of JAK2V617F mutation was determined by using capillary electrophoresis. The results indicated that the JAK2V617F mutation was detected in 59 out of 98 patient with ET, 18 of whom were homozygous mutation.

[Prevalence and clinical significance of FLT3 mutations in acute promyelocytic leukemia].

Objective: To evaluate the prevalence of Fms-Like tyrosine kinase 3 (FLT3) mutations including internal tandem duplication (ITD) of juxtamembrane region and point mutation in the second tyrosine kinase domain (TKD) in acute promyelocytic leukemia (APL) and its clinical significance.

Methods: Bone marrow mononuclear cells from 160 newly diagnosed APL patients were analyzed. Polymerase chain reaction (PCR) was used to detect FLT3-ITD mutations, FLT3-ITD positive samples were further analyzed for the ITD allelic ratio (ITD-AR, mutant-wild type ratio).

[Expression of JAK2V617F and MPLW515L/K mutation in 30 suspected cases of early myeloproliferative disorders].

Objective: To investigate the prevalence of JAK2V617F and MPLW515L/K mutation in patients with slightly elevated platelets (BPC) or hemoglobin (Hb) not meeting the criteria of polycythemia vera (PV) or essential thrombocythemia (ET).

Methods: Genomic DNA from bone marrow or blood mononuclear cells was screened with allele specific polymerase chain reaction (AS-PCR) for JAK2V617F and MPLW515L/K mutation. The history of thrombosis was assessed retrospectively by patients files.

[Experimental study on IL-2- and IL-15 application in allogeneic hematopoietic stem cell transplantation].

Objective: To explore the impact of IL-2- and IL-15-activated donor natural killer (NK) cell infusion on graft-versus-host-disease (GVHD) and graft-versus-leukemia (GVL) effect post allogeneic hematopoietic stem cell transplantation (allo-HSCT).

Methods: The C57BL/6 mice splenic NK cells were selected by microbeads, and then expanded in the media containing IL-2 and IL-15. The killing activity of NK cells was detected.

[Role of CD28/CTLA-4 co-stimulators in immune pathophysiology of aplastic anemia].

Objective: To explore the possible role of CD28/CTLA-4 co-stimulators in immune pathophysiology of acquired aplastic anemia(AA).

Methods: By FACS, the percentages of CD28, CTLA-4 expressing CD3+ CD4+ T cells and the level of Th1, Th2 in bone marrow were detected in 23 AA patients at active phase, 10 at recovery phase and 15 normal controls. The relationship between the co-stimulators, Th1, Th2, and absolute neutrophil counts (ANC) was evaluated.

[Study on relationship between pretransplantation host thymic recent output function and prognosis in HLA-matched sibling hematopoietic stem cell transplantation].

Objective: To study the relationship between pretransplantation host thymic recent output function and prognosis in HLA-matched sibling bone marrow transplantation (MSD-BMT) and determine whether pretransplantation host thymic recent output function can act as a marker for predication of prognosis after HSCT.

Methods: T-cell receptor excision circle (TREC) in DNA of pretransplantation peripheral blood mononuclear cells from 64 patients underwent MSD-BMT was detected by real-time quantitative PCR. The content of TREC in 70 normal donors was detected as well.

[A study of T-cell reconstitution after HLA-matched sibling bone marrow transplantation in leukemia patients].

Objective: To study T-cell reconstitution by the assessment of T-cell receptor excision circle (TRECs) and T-cell receptor clonal repertoire after HLA-matched sibling bone marrow transplantation (MSD-BMT) in leukemia patients and try to reveal the rule of T-cells repopulation in MSD-BMT.

Methods: Real-time quantitative PCR detection of TRECs was carried out in the DNA of peripheral blood mononuclear cells from 23 leukemia patients who underwent MSD-BMT. The content of TRECs in 70 normal donor individuals was also detected to determine the normal range of TRECs in healthy subjects.

[Prediction of T-cell immune reconstitution by investigating T cell receptor excision circle and T-cell receptor beta-chain variable region clonal repertoire in leukemia patients after allogeneic hematopoietic stem cell transplantation].

Objective: To predict T-cell immune reconstitution by investigating T cell receptor excision circles (TREC) and T-cell receptor beta-chain variable region (TCRBV) clonal repertoire in leukemia patients after allogeneic hematopoietic stem cell transplantation (allo-HSCT).

Methods: Real-time quantitative PCR was used to detect the TREC in 43 leukemia patients undergoing matched sibling donor bone marrow transplantation (MSD BMT), matched unrelated donor (MUD) BMT, or haploidentical-stem cell transplantation (HID-SCT), and in 70 normal individuals. RT-PCR was used to amplify 24 subfamily genes of T-cell receptor beta-chain variable region (TCRBV) in 24 of the 43 patients and 5 normal donors as control.

[The molecular characteristics of T-cell immune reconstitution in leukemia patients after allogeneic hematopoietic stem cell transplantation].

Objective: To study the molecular characteristics of CDR3 repertoires of T cell receptor beta chain variable region (TCRBV) of T lymphocytic clones in leukemia recipients after allogeneic hematopoietic stem cell transplantation ( allo-HSCT).

Methods: RT-PCR was used to amplify 24 subfamily genes of TCRBV from peripheral blood (PB) lymphocytes in twenty-four leukemia patients underwent three kinds of allo-HSCT and in five normal donors as control. The PCR products were further analyzed by genescan to evaluate the clonality of BV subfamily and characteristics of CDR3 and calculate usage rate of BV subfamily.