Objective: To assess the clinical characteristics and prognosis of patients with SIL-TAL1-positive T-cell acute lymphoblastic leukemia (T-ALL) . Methods: The clinical data of 19 SIL-TAL1-positive T-ALL patients admitted to the First Affiliated Hospital of Soochow University between January 2014 and February 2022 were retrospectively computed and contrasted with SIL-TAL1-negative T-ALL patients. Results: The median age of the 19 SIL-TAL1-positive T-ALL patients was 15 (7 to 41 years) , including 16 males (84.2%) . SIL-TAL1-positive T-ALL patients had younger age, higher WBC, and hemoglobin compared with SIL-TAL1-negative T-ALL patients. There was no discrepancy in gender distribution, PLT, chromosome abnormality distribution, immunophenotyping, and complete remission (CR) rate. The 3-year overall survival (OS) was 60.9% and 74.4%, respectively (HR=2.070, P=0.071) . The 3-year relapse-free survival (RFS) was 49.2% and 70.6%, respectively (HR=2.275, P=0.040) . The 3-year RFS rate of SIL-TAL1-positive T-ALL patients was considerably lower than SIL-TAL1-negative T-ALL patients. Conclusion: SIL-TAL1-positive T-ALL patients were connected to younger age, higher WBC, higher HGB, and poor outcome.
Adolescent ; Adult ; Humans ; Male ; Young Adult ; Chromosome Aberrations ; Oncogene Proteins, Fusion/genetics* ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; Prognosis ; Recurrence ; Retrospective Studies ; T-Cell Acute Lymphocytic Leukemia Protein 1/genetics* ; T-Lymphocytes ; Female ; Child

This study was aimed to investigate the expression level of CIAPIN1 mRNA in leukemia patients and explore its significance in leukemias. The fresh bone marrow was collected from 112 newly diagnosed leukemia patients, the total RNA was extracted by means of TRIzoL, the cDNA was synthesized, the expression of CIAPIN1 mRNA was detected by real-time quantitative PCR using β-actin as internal reference; 10 normal healthy persons were selected as controls. The results showed that the expression of CIAPIN1 mRNA was statistically higher in acute myeloid leukemia (AML), acute lymphocytic leukemia (ALL) and chronic phase chronic myeloid leukemia (CML) patients than that in normal persons (p < 0.05); but there was no statistical difference between chronic lymphocytic leukemia (CLL) and normal persons (p > 0.05). It is concluded that the CIAPIN1 gene higher expresses in MNC of newly diagnosed leukemia patients, up-regulation of CIAPIN1 expression may play an important role in pathogenesis of leukemia.
Adult ; Bone Marrow Cells ; metabolism ; Case-Control Studies ; Female ; Gene Expression Regulation, Leukemic ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; Leukemia ; genetics ; metabolism ; pathology ; Leukemia, Lymphocytic, Chronic, B-Cell ; genetics ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Young Adult

The aim of this study was to investigate the nucleotide sequence of one distinct fusion transcript of sil-tal1 in childhood T-ALL. The PCR product was cloned into plasmid vector and then sequenced. Genomic DNA was analyzed with PCR using the designed primer pairs representing distinct sequences. The product was sequenced and analyzed with database. The results indicated that 4 different fusion transcripts were detected at cDNA level, in which a part of exons or introns of sil are reserved respectively, and some additions and deletions existed. After analyzing genomic DNA sequence of leukemic cells, the breakpoint in gene sil of this case was proved to be different at DNA level from references. Hence, the sil-tal1 rearrangement was defined to be a new type. It is concluded tal1 rearrangement of leukemic cells in this case is a new type, which expresses classical and at least 3 variant fusion transcripts, presumably caused by extraordinary mechanisms of splicing and transcription in leukemic stem cells.
Base Sequence ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; Child ; DNA Breaks ; DNA, Neoplasm ; genetics ; Humans ; Leukemia-Lymphoma, Adult T-Cell ; genetics ; Male ; Oncogene Proteins, Fusion ; genetics ; Proto-Oncogene Proteins ; genetics ; T-Cell Acute Lymphocytic Leukemia Protein 1 ; Transcription Factors

OBJECTIVETo characterize the time course of spontaneous differentiation of in vitro cultured human embryonic stem cells (hESCs) into hematopoietic cells to provide experimental evidence for induction of hematopoietic commitment of hESCs.

METHODSIn human embryoid bodies (hEBs) derived from spontaneous differentiation of chESC3, a hESC cell line we established previously, the expressions of such genes as KDR, Bmi1, Scl and gata2 were detected by RT-PCR every other day during the 12-day differentiation to monitor the process of the hematopoiesis. The hematopoietic stem cell marker CD34 was examined using flow cytometry to evaluate the efficiency of hematopoietic differentiation of the cells on days 6, 8, 10 and 12. The spontaneously differentiated hESCs were seeded in the hematopoietic colony culture system to study the hematopoietic colony forming ability. Immunocytochemical staining for CD45 was performed on the hEBs to examine the emergence of mature hematopoietic cells.

RESULTSThe expressions of the hematopoietic stem cell-related genes KDR and Bmi-1 were detected in the hESCs, and on days 4 to 6, the two genes were upregulated with prolonged cuture of the hEBs. Scl and gata2 gene expressions were detected since 6-8 days of culture and maintained high expressions till day 12. Flow cytometry revealed a gradual increase in CD34-positive cells in the culture, with positivity rates on days 6, 8, 10, and 12 of (1.4-/+0.4)%, (3.4-/+1.3)%, (5.5-/+2.2)%, and (5.1-/+1.7)%, respectively. The numbers of CD43-positive cell colonies on days 6, 8, 10, and 12 were 0, 7-/+2, 37-/+11, and 89-/+29 in each 10(5) cells, respectively. Immunocytochemical staining identified CD45-positive cells on days 10, 12, 15, and 18 in the cell colonies, with the positive cell numbers of 0, 40.5-/+15.09, 178.6-/+55.89, and 253.0-/+52.04, respectively.

CONCLUSIONThe hESCs undergo spontaneous hematopoietic differentiation in 3 stages, including the differentiation into germ layer-specific cells (days 6-8), expansion period of the hematopoitic progenitors (days 8-12), and maturation of the hematopoietic cells (after day 15).

Animals ; Antigens, CD34 ; metabolism ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; Cell Culture Techniques ; Cell Differentiation ; Embryonic Stem Cells ; cytology ; metabolism ; GATA2 Transcription Factor ; genetics ; Gene Expression Regulation ; Hematopoietic Stem Cells ; cytology ; metabolism ; Humans ; Mice ; Nuclear Proteins ; genetics ; Polycomb Repressive Complex 1 ; Proto-Oncogene Proteins ; genetics ; Repressor Proteins ; genetics ; T-Cell Acute Lymphocytic Leukemia Protein 1 ; Time Factors

OBJECTIVEIn childhood acute lymphoblastic leukemia (ALL), cytogenetics plays an important role in diagnosis, allocation of treatment and prognosis. On the basis of the conventional cytogenetic analysis, molecular methods have improved pediatric hematologists/oncologist's ability to accurately and rapidly perform risk-stratification on patients with childhood ALL during the last few years. The aim of the present study was to assess the demography of cytogenetic abnormalities in childhood ALL.

METHODThe study subjects consisted of 124 newly diagnosed ALL patients younger than 16 years of age, who were diagnosed at the Department of Pediatric Hematology/Oncology, Soochow University Children's Hospital. The diagnosis and FAB subtypes of ALL was determined by Wright-Giemsa-stained bone marrow smears and cytochemical staining. Immunophenotyping of the bone marrow samples was performed by flow cytometry. Multiplex polymerase chain reaction (Multiplex PCR) analysis was performed to detect the 29 most common leukemia translocations for routine molecular diagnostic hematopathology practice, and complement the information gained from conventional cytogenetic analysis.

RESULTSCytogenetic analysis was successful in 112 of 124 children with ALL. Sixty-eight (60%) of them had clonal chromosomal abnormalities. Numerical imbalances consisted of hyperdiploid (> 47 chromosomes, 36 cases), hypodiploid (< 46 chromosomes, 14 cases), pseudodiploidy (18 cases). Chromosomal translocations were observed in 13 patients by conventional cytogenetic analysis. Three cases were found positive for 4; 11 translocation, 3 cases for 9; 22 translocation, 1 case for 1; 19 translocation and 6 cases for other rare translocations. Multiplex-PCR analysis detected 116 of the 124 ALL patients. Thirteen cases of TEL-AML1, 10 cases of rearrangement in the MLL gene, 4 cases of E2A-PBX1, 4 cases of E2A-HLF, 3 cases of BCR-ABL, 2 cases of TLS-ERG, 32 cases of HOX11 were detected by Multiplex PCR in B-lineage leukemias. SIL-TAL1 had been found in 4 of 7 of T-lineage leukemias.

CONCLUSIONSSixty-eight cases of ALL showed chromosomal aberrations. Multiplex PCR positivity was detected in 59 (50%) of the 116 ALL patients studied. Multiplex PCR combined with chromosomal analysis uncovered chromosomal abnormalities in 95 of 124 (77%) of ALL patients and supplemented each other in detecting chromosomal abnormalities.

Adolescent ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; Child ; Child, Preschool ; Chromosome Aberrations ; Core Binding Factor Alpha 2 Subunit ; genetics ; Cytogenetic Analysis ; DNA-Binding Proteins ; genetics ; Female ; Fusion Proteins, bcr-abl ; genetics ; Gene Fusion ; genetics ; Homeodomain Proteins ; Humans ; Immunophenotyping ; methods ; Infant ; Karyotyping ; Male ; Myeloid-Lymphoid Leukemia Protein ; genetics ; Oncogene Proteins, Fusion ; genetics ; Polymerase Chain Reaction ; Pre-B-Cell Leukemia Transcription Factor 1 ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; Proto-Oncogene Proteins ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; T-Cell Acute Lymphocytic Leukemia Protein 1 ; Translocation, Genetic

The prognostic significance of multidrug resistance (MDR) gene expression is controversial. We investigated whether multidrug resistance gene 1 (MDR1), multidrug resistance-related protein (MRP) and lung resistance protein (LRP) mRNA expression are associated with outcomes in acute leukemia patients. At diagnosis we examined MDR1, MRP and LRP mRNA expression in bone marrow samples from 71 acute leukemia patients (39 myeloid, 32 lymphoblastic) using nested RT-PCR. The expression of each of these genes was then expressed as a ratio in relation to beta-actin gene expression, and the three genes were categorized as being either 0, 1+, 2+ or 3+. MDR1, MRP and LRP mRNA expression was detected in 23.9%, 83.1% and 45.1 %, respectively. LRP mRNA expression was significantly associated with resistance to induction chemotherapy in acute leukemia patients, and in the AML proportion (p=0.02 and p=0.03, respectively). MRP and high MDR1 mRNA expression was associated with poorer 2-yr survival (p=0.049 and p=0.04, respectively). Patients expressing both MRP and LRP mRNA had poorer outcomes and had worse 2-yr survival. The present data suggest that MDR expression affects complete remission and survival rates in acute leukemia patients. Thus, determination of MDR gene expression at diagnosis appears likely to provide useful prognostic information for acute leukemia patients.
Vault Ribonucleoprotein Particles/*genetics ; Survival Rate ; RNA, Neoplasm/genetics ; RNA, Messenger/genetics ; Prognosis ; Neoplasm Proteins/*genetics ; Multidrug Resistance-Associated Proteins/*genetics ; Middle Aged ; Male ; Leukemia, Myelocytic, Acute/drug therapy/genetics/mortality ; Leukemia, Lymphocytic, Acute/drug therapy/genetics/mortality ; Leukemia/drug therapy/*genetics/mortality ; Infant ; Humans ; *Genes, MDR ; Gene Expression ; Female ; Child, Preschool ; Child ; Base Sequence ; Aged ; Adult ; Adolescent

OBJECTIVE: To analyze the polymorphisms of glycosylphosphatidylinositol-specific phospholipase D (GPI-PLD) gene exon 14, GPI-PLD activity of leucocyte in the peripheral blood,and the relationship in leukemia patients of Han nationality in Hunan. METHODS: Both 96 leukemia patients and 96 healthy persons of Han nationality in Hunan were researched [including 48 acute non-lymphocytic leukaemia (ANLL) patients as group A, 31 acute lymphoblastic leukaemia (ALL) patients as group B, 12 chronic granulocytic leukaemia (CML) patients as group C, 5 chronic lymphocytic leukaemia (CLL) patients as group D]. The polymorphisms were analyzed by PCR-SSCP and sequencing;. and GPI-PLD activities were determined by GPI-anchored placental alkaline phosphatase (PLAP) as substrate and triton-X114 partioning. RESULTS: There were four variations in the coverage of GPI-PLD gene exon 14 of leukemia patients and healthy persons. The codons of variation were: 1257 C-->T, 1298 T-->C, 1218 C-->A, 1257 C-->A. The total various frequency in leukemia patient and healthy person, which was determined by SSCP, was 28.12% and 20.83%. On the basis of the percentage of GPI-anchored PLAP conversion, the leucocyte GPI-PLD activities of the 96 leukemia patients were measured. Compared with the 96 healthy controls, the leukocyte GPI-PLD activites of ANLL and CLL patients were significantly increased; the acticities of ALL and CML patients were significantly reduced. CONCLUSION Leukocyte GPI-PLD gene in the peripheral blood, which belongs to healthy persons and leukemia patients of Han nationality in Hunan, is polymorphism. The leukocyte GPI-PLD activities in the four groups are remarkable.
Adolescent ; Adult ; Aged ; Base Sequence ; Exons ; genetics ; Female ; Humans ; Leukemia, Lymphocytic, Chronic, B-Cell ; genetics ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Molecular Sequence Data ; Phospholipase D ; genetics ; metabolism ; Point Mutation ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Single-Stranded Conformational ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics

OBJECTIVEHematologic relapse remains the greatest obstacle to the cure of acute lymphoblastic leukemia (ALL), especially T-lineage acute lymphoblastic leukemia (T-ALL) in children. Recent studies have shown that patients with increased risk of relapse can be identified by measuring residual leukemic cells, called minimal residual disease (MRD), during clinical remission. Current polymerase chain reaction (PCR) methods, however, for measuring MRD are cumbersome and time-consuming. To improve and simplify MRD assessment, the author developed a real-time quantitative PCR (RQ-PCR) assay for the detection of leukemic cells that harbor the tal-1 deletion. In addition, the author discussed the significance of MRD levels at different stages in treatment and prognosis of children with T-ALL.

METHODSA total of 50 consecutively enrolled patients with T-ALL were analysed for detection of leukemic cells harboring the most common tal-1 deletion. Serial dilutions of leukemic DNA were studied to find the sensitivity of detection with RQ-PCR assay. The MRD of 28 samples in clinical remission from 10 patients were quantified by RQ-PCR assay and limiting dilution assay. The results detected by both methods were compared statistically with correlation analysis.

RESULTS(1) A total of 10 patients presented tal-1 deletion involving the sildb1 breakpoint rearranged to tal1db1 in 50 cases with T-ALL. The breakpoints of relapsed samples are the same as those of the corresponding diagnostic samples; (2) The RQ-PCR assay had a sensitivity of detection of one leukemic cell among 100,000 normal cells. In 24 samples, MRD levels > 10(-5) could be detected with both methods. The percentages of leukemic cells measured by the two methods correlated well (r = 0.898, P < 0.001); (3) The MRD levels of 3 patients out of the 8 cases undergoing disciplinary regimen were over 10(-4) at the end of induction chemotherapy. They all relapsed in bone marrow during chemotherapy. The higher the MRD levels, the earlier the relapse. The other 5 patients with MRD levels < 10(-4) had been relapse-free survival (RFS) for 4-59 months, one of whom with increased MRD levels > 10(-4) for twice at the continuation stage had been RFS for 27 months till now.

CONCLUSIONSThe sildb1-taldb1 deletion presents in 20% of T-ALL, and is an ideal PCR marker for its specificity, uniform and stability; The tal-1 RQ-PCR can be used for the rapidly, sensitively and accurately quantitative assessment of MRD in T-ALL with the tal-1 deletion. MRD levels at different stages of chemotherapy have different significance in prognosis and treatment.

Adolescent ; Base Sequence ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; Child ; Child, Preschool ; Female ; Gene Deletion ; Humans ; Male ; Molecular Sequence Data ; Neoplasm, Residual ; diagnosis ; Polymerase Chain Reaction ; methods ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; genetics ; mortality ; Prognosis ; Proto-Oncogene Proteins ; genetics ; T-Cell Acute Lymphocytic Leukemia Protein 1

Basic Helix-Loop-Helix Transcription Factors ; metabolism ; Chromosome Aberrations ; DNA Nucleotidylexotransferase ; metabolism ; Humans ; Immunoglobulin Light Chains ; metabolism ; Immunoglobulin Light Chains, Surrogate ; Lymphoma, B-Cell ; genetics ; metabolism ; pathology ; Lymphoma, T-Cell ; genetics ; metabolism ; pathology ; Membrane Glycoproteins ; metabolism ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; metabolism ; pathology ; Proto-Oncogene Proteins ; metabolism ; T-Cell Acute Lymphocytic Leukemia Protein 1

The aims of this study were to estimate the incidences of BCR/ABL, MLL, TEL/AML1 rearrangements, and p16 deletions in childhood acute lymphoblastic leukemia (ALL), to identify new abnormalities, and to demonstrate the usefulness of interphase fluorescence in situ hybridization (FISH). We performed G-banding analysis and FISH using probes for BCR/ABL, MLL, TEL/AML1 rearrangements, and p16 deletions on 65 childhood ALL patients diagnosed and uniformly treated at a single hospital. Gene rearrangements were identified in 73.8% of the patients using the combination of G-banding and FISH, while the chromosomal abnormalities were identified in 49.2% using G-banding alone. Gene rearrangements were disclosed by FISH in 24 (72.7%) of 33 patients with normal karyotype or no mitotic cell in G-banding. Among the gene rearrangements detected by FISH, the most common gene rearrangement was p16 deletion (20.3%) and the incidences of others were 14.1% for TEL/AML1, 11.3% for MLL, and 1.8% for BCR/ABL translocations. Infrequent or new aberrations such as AML1 amplification, MLL deletion, ABL deletion, and TEL/AML1 fusion with AML1 deletion were also observed. We established the rough incidences of gene rearrangements in childhood ALL, found new abnormalities and demonstrated the diagnostic capability of interphase FISH to identify cryptic chromosome aberrations.
Adolescent ; Child ; Child, Preschool ; *Chromosome Aberrations ; Chromosome Banding ; DNA-Binding Proteins/*genetics ; Female ; Fusion Proteins, bcr-abl/*genetics ; Gene Deletion ; *Gene Rearrangement ; Humans ; In Situ Hybridization, Fluorescence ; Infant ; Interphase ; Leukemia, Lymphocytic, Acute/*genetics ; Male ; Oncogene Proteins, Fusion/*genetics ; Protein p16/*genetics ; Proto-Oncogenes/*genetics ; Transcription Factors/*genetics ; Treatment Outcome