LNK Gene Single Nucleotide Polymorphisms and Acute Leukemia Susceptibility / 中国实验血液学杂志

<p><b></b>To investigate the relationship between the LNK(SH2B3) gene single nucleotide polymorphism and risk of acute leukemia (AL) in Chinese population.</p><p><b>METHODS</b>The bone marrow and peripheral blood samples from 31 cases of acute lymphoblastic leukemia, 70 cases of acute myeloid leukemia and 130 healthy persons as the controls were collected. Genotype of LNK SNP Rs3184504(c.784T>C) and Rs78894077(c.724C>T) were determined by PCR-RFLP, and were confirmed by gel electrophoresis and sequencing. The NB4, THP-1 and Raji leukemia cell line models were cultured, the leukemia cell line LNK Rs3184504 and Rs78894077 polymorphism were detected by using direct sequencing.</p><p><b>RESULTS</b>The CC genotype frequencies of Rs3184504 SNP were higher in ALL and AML patients than those in control (P<0.01), but there was no different between the groups in AML and ALL. The frequency of LNK gene Rs3184504 C allele was higher in AL as compared with control (P<0.01). The LNK gene Rs78894077 locus genotype distribution was not significantly different between the AL and the normal control group (P>0.05). Both Rs3184504 and Rs78894077 sites were detected as CC genotype in NB4, THP-1 and Raji cells.</p><p><b>CONCLUSION</b>The persons carrying C allele of LNK gene Rs3184504 are more prone to develop acute leukemia.</p>

High Mobility Group Chromosal Protein N2 Is Expected to be as A Target of Cellular Immunetherapy in Leukemia and Tumors / 中国实验血液学杂志

Recently, chimeric antigen receptors T cells (CAR T) have made a breakthrough in the treatment of lymphoma and leukemia, open a new path for the tumor cellular immunetherapy. It is the key for CAR T to take the gene which can identify the CD19 antigen of lymphoblastic leukemia into lymphocytes, enable it to kill leukemia cells with specific cell-surface loci. The same principle also applies to other aspects, if we find specific target genes of lymphocytes. Recent studies have found that high mobility group protein N2 (high mobility group chromosal protein N2, HMGN2) is the excellent target of tumor-associated antigen in lymphocytes, is the antitumor effector molecule of CD8(+) T cells, which has the ability of trends and specific identify/binding in myeloid leukemia, breast cancer, cervical cancer and other tumor cells. HMGN2 is expected to be used for the preparation of specific identification of tumor lymphocytes and to treat more leukemia and tumors. This article focuses on the strucure and function of HMGN and the chemotaxis and antitumor effect of HMGN2 in leukemia and tumors.

Molecular Mechanism and Malignant Clonal Evolution of Multiple Myeloma / 中国实验血液学杂志

Almost all patients with multiple myeloma (MM) have chromosomal translocation which can result in genetic variation. There are mainly five types of chromosomal translocations, involving the IGH gene translocation to 11q13 (CCND1), 4p16 (FGFR/MMSET), 16q23 (MAF), 6p21 (CCND3) and 20q11 (MAFB). It is possible that all IGH translocations converge on a common cell cycle signal pathway. Some MM develops through a multistep transformation from monoclonal gammopathy of undetermined significance (MGUS) to smoldering MM (SMM) and eventually to MM and plasma cell leukemia (PCL). Similarly to what Darwin proposed in the mid-19th century-random genetic variation and natural selection in the context of limited resources, MM clonal evolution follow branching and nonlinear mode. The failure of MM treatment is usually related with the minimal subclone which is hardly found at newlydiagnosed.

Treatment of lymphoblastic leukemia with CD19-specific modified chimeric antigen receptor T cells / 中国实验血液学杂志

Bioengineered T cells, which are the genetically manipulated T cells to express chimeric antigen receptor T Cell (CAR T) against leukemia-associated specific antigens, were applied to treat acute and chronic lymphocytic leukemia with CAR T. CAR T cells combined with cell-surface binding site and anti-CD19 chimeric antigen receptor can treat diseases through T cells transfection. CAR T cells can recognize the CD19 antigen on B cells with specific cell-surface loci. CAR T cells can proliferate by 1000 times and differentiate in vivo by the CD19 antigen stimulation, therefore, kill the acute and chronic lymphocytic leukemia cells effectively. This article briefly reviews the CAR T cells and the effect of CAR T cells on acute and chronic lymphoblastic leukemia.

Identification of splenic marginal zone lymphoma from B lymphoproliferative disorders by flow cytometry / 中国实验血液学杂志

The splenic marginal zone lymphoma (SMZL) is a relatively rare chronic B lymphoproliferative disease, which primarily manifest increase of peripheral blood lymphocyte count and/or scale, and splenomegaly, while the peripheral superficial lymph nodes are often not swollen. Therefore, the splenectomy are usually needed to confirm the diagnosis, but the majority of patients could not accept such management, resulting in early difficult diagnosis. This study was purposed to explore the more prior way for diagnosis based flow cytometry (FCM). Six patients with suspected diagnosis of SMZL were used as research objects, 10 healthy bone marrow donors and 10 cases of chronic lymphocytic leukemia (CLL), 3 cases of hairy cell leukemia (HCL), 3 cases of lymphatic plasma cell lymphoma/Waldenströ's macroglobulinemia (LPL/WM) were selected as control. The immunophenotype of bone marrow cells were analyzed and compared by FCM using a panel of antibodies including CD45, CD5, CD10, CD19, CD20, CD22, CD23, CD25, CD103, CD11c, CD123, κ,λ, Cyclin D1, and combined with bone marrow cell morphology. The results indicated that 6 cases of suspected SMZL showed a large increase of lymphocytes and splenomegaly. Because absence of peripheral lymphadenopathy, 6 patients did not suffer from lymph node biopsy, only 1 patient underwent diagnostic splenectomy. The immunophenotypes of bone marrow in patients and controls were analyzed by FCM, as a result, except for the healthy donors, varying degrees of abnormal mature B cell clones were found in bone marrow of all patients, and the further differentiation from other B-cell tumors was performed through CD5, CD10 expression and combination with other B-cell phenotype. All 6 cases of SMZL patients expressed CD19(+) and CD20(+), but CD10 expression was negative, 4 patients expressed CD5(-), 2 patients expressed CD5(+). The expressions of CD23, CD38, ZAP-70, CD11c, CD103, CD123, Cyclin D1 were negative. The morphological examination of bone marrow cells showed velutinous abnormal lymphocytes. Combined with clinical characteristics, 6 patients were diagnosed as SMZL, 1 patient suffered from splenectomy because of concurrent hypersplenism, and this postoperative pathologic examination confirmed the patient with SMZL. Ten cases of CLL mainly expressed CD5, CD23; 3 cases of HCL had more typical morphology of "hair like" in addition to CD11c, CD103 and CD123 positive; 3 cases of LPL/WM had significantly increased light chain restriction expression, IgM, plasmacytoid lymphocytes. It is concluded that the FCM immunophenotype analysis can be used as a powerful tools for clinical diagnosis of SMZL.

Role of LNK gene mutation in pathogenesis of myeloproliferative neoplasms-review / 中国实验血液学杂志

Myeloproliferative neoplasms ( MPN ) is a class of clonal hematopoietic stem cell disease. Studies found that the JAK-STAT signaling pathway is closely related to the pathogenesis of MPN. The lymphocyte-specific adaptor protein (LNK) gene negatively regulates Janus kinase-signal transducer and activator of transcription (JAK-STAT) signaling and may play an important role in the pathogenesis of MPN. Especially in JAK2 mutation-negative MPN, LNK gene specific mutations may be the key to cause MPN subtypes. Certain single nucleotide polymorphism of LNK gene regulation of hematopoietic cells in different directions may also be important influence factors of MPN performance for different subtypes. LNK gene functional changes lead to abnormal activation of the JAK-STAT signaling pathway, and may be a new mechanism of MPN. In this review, the role of LNK gene in MPN pathogenesis is briefly summarized.

Role of signal transducer and activator of transcription 3 in the development of tumors / 中国实验血液学杂志

Signal transducer and activator of transcription 3 (STAT3) take part in cell proliferation, differentiation, survival, apoptosis, transformation, cellular immunity and some other important physiological and pathological processes. Among STAT3 signaling pathways, the JAK-STAT signaling pathway has been comprehensively studied. Abnormal activation of STAT3 is frequently detected in various tumors, and the abnormal activation is closely related with the tumorigenesis. Recent studies have found that mutations and several specific genotypes of single nucleotide polymorphisms in STAT3 gene may be involved in tumor formation, also suggesting the important role of STAT3 in tumor biology. In this review, the role of STAT3 in the development of tumors is briefly summarized.

Inherited paternal antigens induce pregnancy thrombocytopenia / 中国实验血液学杂志

Objective of this study was to investigate the correlation of body-carried inherited paternal antigens (IPA) in one mother after delivery with pregnancy thrombocytopenia. The changes of platelet (Plt) count in the mother who delivered 2 years ago and her child who is now one year's old were detected, routine tests included Helicobacter pylori, CMV, EBV, parvovirus and other herpes virus's infection were carried out. Eight insertion or deletion sites (InDel) SNP with strong polymorphisms in Chinese population was selected to detect IPA from a genomic library, then primers were designed, the nested PCR and real-time quantitative PCR were used to detect 54 healthy mother-child pairs, the obtained average value was taken as the control, finally two InDel polymorphism sites between mother and child were used to identify the mother/child microchimerism. The IPA of the mother were examined at 4 time points. The results showed that the Plt level of the mother who had suffered thrombocytopenia since 20 weeks after pregnancy reduced to 10 × 10(9)/L. After using gamma globulin, the Plt count increased gradually, but the Plt count decreased rapidly when withdrawal. This patient did not have the infections of virus and Helicobacter pylori. IPA average value of 54 cases were from 10(-5) to 10(-4). At 67 d after delivery, the Plt count of the mother was 14 × 10(9)/L, IPA was 3.45 × 10(-3), which was 30 times higher than the normal. In one month after treatment the IPA was 1.3 × 10(-4) (Plt 256 × 10(9)/L), 5 months later it was 1.2 × 10(-4) (Plt 158 × 10(9)/L), and 6 months later it was 1.5 × 10(-4) (Plt 325 × 10(9)/L). When IPA reached the normal level, the Plt count returned to normal. Her child suffered thrombocytopenia (4 × 10(9)/L) one month after he was born, then recovered after high-dose gamma globulin therapy. It is concluded that abnormal high level IPA may lead to pregnancy thrombocytopenia.

Relationship between V617F mutation and 46/1 haplotype in JAK2 gene in patients with chronic myeloproliferative diseases and frequencies of 46/1 haplotype in different Chinese nationalities / 中国实验血液学杂志

Somatic gene V617F mutation in JAK2 is a critical molecular and biological indicator to diagnosis of chronic myeloproliferative disease (MPD). This study was aimed to investigate the genetic background of V617F mutation in 46/1 gene haplotype in Chinese MPD patients, and the frequencies of 46/1 gene haplotype and V617F mutation in three nationalities of Chinese populations. Peripheral blood or bone marrow samples of 150 V617F mutation positive MPD patients, 123 V617F mutation negative MPD patients, 124 healthy Han individuals, 395 healthy Tibetan individuals and 315 healthy Yugu individuals were collected. The allele-specific multiplex PCR method was established, the presence or absence of V617F mutation, the presence or absence of 46/1 haplotype, and the relationship between V617F and 46/1 haplotype were easily identified by agarose gel image. The results showed that the V617F mutation located in the 46/1 haplotype of 88 cases (58.67) among 150 V617F-positive MPD cases. In 814 Chinese healthy individuals including Han, Tibetan, Yugu nationalities, the frequency of the 46/1 gene haplotype was 38.37 without difference in the frequency among different nationalities, and no V617F mutation was found in Chinese healthy populations, The frequency of the 46/1 gene haplotype was 43.09 in V617F mutation negative MPD patients and was 69.33 in V617F mutation positive MPD patients, the latter was obviously higher than former and than that in healthy Han individuals. In conclusion, a multiplex PCR method has been developed that is simple and useful to identify V617F mutation in JAK2 gene and its relationship to the 46/1 haplotype. In more than half of Chinese V617F-positive MPD patients, the V617F mutation locates in 46/1 haplotype in JAK2. The frequencies of 46/1 haplotype are statistically insignificant among Han, Tibetan and Yugu nationality populations.

Effects of Iron Chelators on Labile lron Pool and Apoptosis Related Genes Ex pression in K562 Cells / 实用儿科临床杂志

Objective To explore the effect of iron chelators on labile iron pool and expression of apoptosis associated genes in cells of K562, an erythroleukemia cell line.Methods K562 cells were incubated at 37 ℃ in RPMI 1640 containing 10% heat-inactived fetal bovine serum in an saturated humidity and 5% CO_2 incubator. K562 cells were incubated with different concentrations of desferro-(xamine(DFO)). The study groups were divided as following: DFO group, iron+DFO group and the control group. Following indices were detected which included apoptosis by flow cytometry (FCM) assay, expression of Rb, c-myc, bax mRNA by RT-PCR. The intracellular LIP was measured with a fluorimetric assay using the metalsensitive probe calcein-AM.Results 1. The viability of K562 cells incubated with different concentrations of DFO was lower than that of control group at 12 h,24 h and 48 h (P