Focal lymphoblastic transformation of chronic myelogenous leukemia develops into erythroid leukemia:A case report

BACKGROUND We present a case of focal lymphoblastic transformation to erythroid leukemia following acute myeloblastic transformation in a patient with chronic myelogenous leukemia(CML)and discuss its mechanism of occurrence and development.CASE SUMMARY The presence of the Philadelphia(Ph)chromosome was identified through karyotype analysis,while the BCR-ABL fusion gene was detected using quantitative real-time polymerase chain reaction of the peripheral blood sample.Fluorescence in situ hybridization was used to detect the expression of the BCRABL gene in the lymphoma.Antigen expression and gene mutations in the primitive cells were detected by flow cytometry.The analysis confirmed the presence of CML along with focal lymphoblastic transformation to erythroid leukemia.Additionally,the patient was found to have secondary erythroid leukemia,along with multiple new gene mutations and abnormalities in complex karyotypes of chromosomes.CONCLUSION Our findings suggest a possible molecular basis for the focal lymphoblastic transformation secondary to myeloblastic transformation in patients with CML.

Detection of Telomerase Activity and the Expression of Telomerase Subunits in the Patients with Acute Myelogenous Leukaemia

Telomerase activity and the expression of telomerase subunits (for example, telomerase reverse transcriptase and telomerase associated protein 1 and telomerase RNA component) of peripheral white blood cells were detected in the patients with acute myelogenous leukaemia (AML) and the correlation between telomerase activity and the expression of telomerase subunits was observed. In 94 peripheral white blood cells from 18 healthy volunteers and 76 patients with AML, including 31 AML at initial presentation, 24 at relapse and 21 at complete remission, the telomerase activity and telomerase subunits mRNA or RNA were detected by PCR ELISA and RT PCR respectively. The results showed that the positive rate of telomerase from patients with AML at initial presentation, at relapse and at complete remission was 74.1 %, 79.2 % and 4.8 % respectively. The positive rate of telomerase reverse transcriptase mRNA from healthy volunteers, AML at initial presentation, AML at relapse and AML at complete remission was 5.6 %, 80.6 %, 83.3 % and 9.5 % respectively. The positive rate of telomerase associated protein 1 mRNA and telomerase RNA component in all samples were 100 %. It was suggested that the up regulation of telomerase activity and the expression of telomerase reverse transcriptase is correlated closely with the occurrence and relapse of AML, so telomerase activity and the expression of telomerase reverse transcriptase may be used to estimate the curative effect and predict relapse of AML. Moreover, the up regulation of telomerase activity is correlated with the expression of telomerase reverse transcriptase significantly.

Correlation of Aurora family and TRPV family gene expression in acute myelogenous leukemia with disease progression

Objective:To study the correlation of Aurora family and TRPV family gene expression in acute myelogenous leukemia with disease progression.Methods: Patients who were diagnosed with acute myelogenous leukemia in Kashgar Prefecture First People's Hospital between January 2014 and September 2017 were selected as the leukemia group of the research, and patients who underwent bone marrow puncture and were without myelogram anomaly in Kashgar Prefecture First People's Hospital during the same time were selected as the control group. The bone marrow tissue was collected to determine the mRNA expression of Aurora family and TRPV family genes, apoptosis genes and metastasis genes.Results: Aurora-A, Aurora-B, Aurora-C, TRPV-5, TRPV-6, GFI-1, Bcl-2, CXCR3, C3AR1,β-arrestin1, MMP2 and MMP9 mRNA expression in bone marrow tissue of leukemia group were significantly higher than those of control group whereas CD40L, Bax, Caspase-9 and Caspase-3 mRNA expression were significantly lower than those of control group. Aurora-A, Aurora-B, Aurora-C, TRPV-5 and TRPV-6 mRNA expression in bone marrow tissue of leukemia group were positively correlated with GFI-1, Bcl-2, CXCR3, C3AR1,β-arrestin1, MMP2 and MMP9 mRNA expression, and negatively correlated with CD40L, Bax, Caspase-9 and Caspase-3 mRNA expression.Conclusion: The high expression of Aurora family and TRPV family genes in acute myelogenous leukemia can promote the proliferation and metastasis of tumor cells during disease progression.

Myeloid sarcoma presenting as a colon polyp and harbinger of chronic myelogenous leukemia

Myeloid sarcoma, also known as granulocytic sarcoma or chloroma is an unusual accumulation of malignant myeloid precursor cells in an extramedullary site, which disrupts the normal architecture of the involved tissue. It is known to occur more commonly in patients with acute myelogenous leukemia and less commonly in those with myelodysplastic syndrome and myeloproliferative neoplasm, such as chronic myelogenous leukemia. The most common sites of involvement include bone, skin and lymph nodes. However, rare cases have been reported in the gastrointestinal tract, genitourinary tract, or breast. Most commonly, a neoplastic extramedullary proliferation of myeloid precursors in a patient would have systemic involvement of a myeloid neoplasm, including in the bone marrow and peripheral blood. Infrequently, extramedullary disease may be the only site of involvement. It may also occur as a localized antecedent to more generalized disease or as a site of recurrence. Herein, we present the first case in the English literature of a patient presenting with an isolated site of myeloid sarcoma arising in the form of a colonic polyp which, after subsequent bone marrow biopsy, was found to be a harbinger of chronic myelogenous leukemia.

Redistribution of Platelet Membrane Glycoprotein IV and Release of Intracellular α-granule Thrombospondin in Patients with Chronic Myelogenous Leukemia

The redistribution of platelet membrane glycoprotein IV (GPIV) and the release of intracellular Q-granule thrombospondin (TSP) were examined and the inhibition of 5-thromboglobulin (&TG) and platelet factor 4 (PF4) in patients with chronic myelogenous leukemia (CML) was observed and quantitation of β-TG and PF4 in sera was conducted. GPIV in inactive platelet from CML was 36080±17010 molecules/platelet as compared with 13190±4810 from the controls (P<0,01), No abnormality was found in the distribution of platelet membrane GPIb and GPIIb/III.(P>0. 05). The GPIV redistribution on active platelet membrane induced thrombin (1U/ml) from CML and healthy donors was 44320132310 and 228001 12700 molecules/platelet respectively (P<0. 01 ). The difference in the release of intracellular Q-granule TSP between CML and the control group was not found (P>0.05). There was no direct correlation between GPIV expression and TSP binding after platelet activation. The high leveIs of β-TG and PF4 in sera inhibited release of intracellular a-granule TSP in vitro. These results indicate that the abnormality of platelet membrane GPIV is a common marker in CML, therefore the specific increase of platelet GPIV in patients with CML may be a useful tool for the diagnosis and monitoring of the platelet dysfunction. The release of interna1 TSP pools is hindered by either β-TG or PF4 in sera.

HIM_(1) AND HIM4, TWO MONOCLONAL ANTIBODIES POTENTIALLY USEFUL FOR AUTOLOGOUS BONE MARROW TRANSPLANTATION IN CHRONIC MYELOGENOUS LEUKEMIA

We developed two complement-fixing MoAbsHIMand HIM(murine)that were specifically reac-tive with chronic myelogenous leukemia (CML) cells.They were capable of fixing human or rabbit com-plement and suitable for CML cells purging of re-mission marrow from CML patients.HIMreactedwith majority leukemic cells form 7 out of 10 CMLpatients by complement-mediated cytotoxicity(C’MC)assay(positive cells 80%—90%),HIMreacted withmajority CML cells from 4 out of 5 CML by C’MCassay(positive cells 80%—90%).Treatment withHIMor HIMand human C’was capable of lysing97% of K562,U937,HL-60 and CML cells in a 20fold excess of unrelated cells by indirect FITC+EBstain.Using limited dilution culture,incubation withHIMand C’produced 1.5 logs inhibition of growthin K562 cells,and 1.9 logs in U937 cells,and withHIMand C’produced 2.9 logs inhibition in HL-60cells and 3.0 logs in U937 cells.Both MoAbs cocktailwas shown 1.8 logs in K562 cells and 3.2 logs in U937cells.They were no suppression on the growth o

Preliminary Research on the p53 Gene Rearrangements in the Evolution of Chronic Myelogenous Leukemia to Blast Crisis

DNA from 36 patients with chronic myelogenous leukemia (CML) at various clinical stages and 6 cases of acute leukemia was investigated for alterations of the p53 gene by Southern blot analysis.Rearrangements of the p53 gene were seen in 3 of 12 (25.00%) cases of blast crisis and accelerated phase (AP) of CML and in only one of 18 chronic phrase (CP),just as has been reported previously. Meanwhile,by restriction fragment length polymorphism (RFLP) analysis the Bgl II site polymorphism in the p53 gene was also found. The frequency in Chinese people detected here was 0.392,which was strikingly higher than that in some other countries(P<0. 001).These results suggested that the alterations of the p53 gene, for example,p53 rearrangements,were probably responsible for the progression of BC in some CML patients, and that the frequency of Bgl II polymorphism in the p53 gene might be related to the population distribution.

Correlated Flow Cytometric Analysis of H－ras p21 and DNA Ploidy in Acute Myelogenous Leukemia

The flow cytometric immunoassay was used to study the correlation between the H-ras oncogene product p21 and the DNA ploidy in 30 de novo cases of acute myelogenous leukemia (AML). The results showed that 17 cases were negative for p21 expression and 13 positive for p21. The patients with positive p21 had higher percentage of bone marrow and peripheral blasts and lower peripheral leukocyte count. The expression of p21 had no influence on the therapeutic effect. Before treatment,DNA diploidy occurred in 18 cases including 13 p21 negative ones,and DNA aneuploidy was revealed in 12 cases including 8 p21 positive ones. Patients with positive p21 or having aneuploidy in complete remission were at risk for early relapse. Our results suggest that p21 may be involved in the process of leukemogenesis and progression in AML.

Research on Marketing Strategies of Product D—a Chronic Myelogenous Leukemia Drug for Pharmaceutical Company A

Objective To put forward some suggestions on the marketing strategies for chronic myelogenous leukemia in a pharmaceutical enterprise.Methods Based on the development status of the pharmaceutical industry and the SWOT analysis of a company’s product,the marketing strategies were formulated to provide theoretical basis for pharmaceutical enterprise to adapt to the new medical reform.Results and Conclusion Nowadays,due to fierce competition,in order to expand the new market,enterprises should implement the strategies of new products,centralized management and professional training.Meanwhile,the effective marketing strategies should be formulated and strictly carried out according to the conditions of the pharmaceutical company.

RNF6 promotes chronic myelogenous leukemia cell proliferation and migration by stabilizing vimentin via multiple atypical ubiquitinations

Chronic myelogenous leukemia(CML)is a malignancy from bone marrow myeloid stem cells mainly driven by the fusion gene BCR-ABL.In addition to BCR-ABL,other genes including RNF6 are also dysregulated in CML cells.1 RNF6,a ubiquitin ligase of the RING family,promotes various cancer cell proliferation,chemoresistance,and tumor growth in vivo by targeting various proteins for ubiquitination and degradation,including SHP1,TLE3,FOXA1,and MAD1.^(2) However,its specific mechanism in CML is not known.

Prolonged chronic phase in chronic myelogenous leukemia after homoharringtonine therapy

Background Homoharringtonine （HHT） is effective in treating late stage chronic myelogenous leukaemia （CML）, but little is known about long term maintenance during complete cytogenetic response. Long term efficacy and toxicity profiles of low dose HHT were evaluated in this study. Methods One hundred and six patients with CML received 1.5 mg/m^2 of HHT alone by continuous daily infusion for seven to nine days every four weeks. Of 79 patients in the control group, 31 were treated with interferon α （IFN-α） and 48 with hydroxycarbamide. For 17 patients who failed to achieve cytogenetic response within 12 months＇ treatment of IFN-α, HHT was administered. Quantitative RT-PCR was used to detect the BCR-ABL mRNA expression in 36 Philadelphia positive CML patients enrolled after 2007. Haematological and cytogenetic responses were evaluated in all patients at the 12th month of follow-up. Long term efficacy was assessed in a follow-up with a median time of 54 months （12 months-98 months）. Results After 12 months of therapy, cytogenetic response rate of the HHT, IFN-α and hydroxycarbamide groups were 39/106, 14/31 and 3/48, and corresponding molecular cytogenetic response rates 6/18, 3/8 and 0. Of the 17 patients who received HHT as salvage treatment, 6 achieved cytogenetic response （3 major）. At the 48 months＇ follow-up, cytogenetic response was maintained in 32/39 patients treated with HHT. Patients who had cytogenetic response in HHT group or treated with IFN-α also showed longer median chronic durations, which were 45 months （12 months-98 months） and 49 months （12 months-92 months） respectively, indicating a longer survival time. Conclusions Low dose HHT alone showed considerable short term and long term efficacy in the treatment of late stage CML. It may also be a good choice for patients who have failed imatinib, IFN-α treatment or haematopoietic stem cell transplantation or cannot afford these treatments.

Arsenic trioxide downregulates the expression of annexin II in bone marrow cells from patients with acute myelogenous leukemia

Background Most patients with acute myelogenous leukemia （AML） suffer from disordered hemostasis. We have previously shown that annexin II （Ann II）, a high-affinity co-receptor for plasminogen/tissue plasminogen activator, plays a central role in primary hyperfibrinolysis in patients with acute promyelocytic leukemia （APL）. The expression of Ann II in cells from patients with major subtypes of AML and the effect of arsenic trioxide （As203） on Ann II expression in AML cells were investigated to determine whether As203-mediated downregulation of Ann II could restore hemostatic stability. Methods A total of 103 patients （48 females and 55 males; age, 19-58 years） were included. Plasma samples were collected before and after treatment as well as after complete remission. Ann II and plasminogen activation were measured in leukemic cells during treatment with 1 pmol/L As203. Results Before AS203 treatment, Ann II mRNA expression （real-time PCR） was the highest in M3 cells （P 〈0.05）, higher in M5 cells than that in M1, M2, M4, and M6 cells （P〈0.001）, and positively correlated with Ann II protein expression （flow cytometry） （r=0.752, P 〈0.01）. Exposure for up to 120 hours to As203 （1 μmol/L） had no significant effect on Ann II protein in M1 and M2 leukemic cells, but decreased Ann II protein expression twofold within 48 hours of exposure in M3 cells （P 〈0.05） and twofold within 96 hours in M5 cells （P 〈0.05）. The rate of plasmin generation was higher in APL, M5, and M4 cells than in M1, M2, and M6 cells. Conclusions As203 may reduce hyperfibrinolysis in AML by downregulation of Ann 11. Furthermore, As2O3 affects more than one form of AML （APL, M4 and M5）, suggesting its potential role in their management.

Severe vulgaris psoriatic patients with acute myelogenous leukaemia and resolution after allogeneic bone marrow transplantation/ peripheral blood stem cell transplantation

Psoriasis is a common disease with a prevalence of up to 2% in the world population.~1 The pathophysiological mechanism of this inflammatory disease has not been determined although it is generally accepted to have a genetic predisposition and be T-cell mediated. The lymphocytes are the target of psoriatic therapeutic strategies. Therapy resolves psoriasis for a time, but induces some severe side effects. Some cases of severe psoriasis, treated with multiple therapeutic regimens for many years, developed leukemia.

High expression of RbAp46 gene in patients with acute leukemia or chronic myelogenous leukemia in blast crisis

The retinoblastoma （Rb） suppressor associated protein 46 （ RbAp46 ） also named retinoblastoma binding protein 7 （RBBP7） was first identified as a protein in HeLa cells that binds to an Rb affinity column. RbAp46 has been shown to be a core component of the mSin3 histone deacetylase （HDAC） complex and NuRD （ a multi-subunit complex containing chromosome-remodeling activity）. 2 RbAp46 is also known as the histone acetyltransferase （HAT） type B subunit

The role of RAS effectors in BCR/ABL induced chronic myelogenous leukemia

BCR/ABL is the causative agent of chronic myelogenous leukemia(CML).Through structure/function analysis,several protein motifs have been determined to be important for the development of leukemogenesis.Tyrosine177 of BCR is a Grb2 binding site required for BCR/ABL-induced CML in mice.In the current study,we use a mouse bone marrow transduction/transplantation system to demonstrate that addition of oncogenic NRAS(NRASG12D)to a vector containing a BCR/ABL^(Y177F)mutant“rescues”the CML phenotype rapidly and efficiently.To further narrow down the pathways downstream of RAS that are responsible for this rescue effect,we utilize well-characterized RAS effector loop mutants and determine that the RAL pathway is important for rapid induction of CML.Inhibition of this pathway by a dominant negative RAL is capable of delaying disease progression.Results from the present study support the notion of RAL inhibition as a potential therapy for BCR/ABL-induced CML.

Inhibitory effects of rapamycin on proliferation of chronic myelogenous leukemia cells and its mechanism

Objective To explore the inhibitory effects of rapamycin on proliferation of chronic myelogenous leukemia (CML) cells and its possible mechanism. Methods The effects of rapamycin at various concentrations on cell proliferation of CML cell line K562 cells were analyzed by MTT. The

Comparison of efficacy and prognostic factors in elderly acute myelogenous patiens with MA or CAG induction chemotherapy regimen

Objective To analyze the difference of efficacy and prognostic factors between MA and CAG induction chemotherapy in elderly acute myelogenous leukemia(AML)patients.Methods From April 2009 to September 2015,103 consecutive hospitalized 60-plus-year-old AML patients were retrospectively analyzed.NPM1

格列卫对K562细胞中miR-146a、miR-29b及3种甲基化酶表达的影响

目的观察BCR/ABL抑制剂格列卫作用后K562细胞中microRNA(miR)-146a及miR-29b的表达水平变化及DNMT1、DNMT3a、DNMT3b3种甲基化酶水平的改变。方法 MTT法检测格列卫作用于K562细胞时的IC50浓度。通过茎环引物法及荧光定量PCR的方法检测miRNAs以及甲基化酶基因的水平。结果格列卫作用于K562细胞时的IC50浓度为40.85μmol/L。格列卫作用后miR-29b表达出现了上升的趋势而3种甲基化酶基因表达水平均有所降低,miR-146a的水平显著升高(P<0.05)。结论格列卫能影响K562细胞中miR-146a、miR-29b和3种甲基化酶的表达水平。

GENE THERAPY USING RETROVIRAL VECTOR OF bcr-abl SPECIFIC MULTI-UNIT RIBOZYMES COULD INHIBIT CML CELL GROWTH AND INDUCE APOPTOSIS

Objective: To investigate the in vitro cleavage ability and effects on apoptosis and cell growth of the bcr-abl fusion gene specific multi-unit ribozymes. Methods: Three fusion point specific ribozymes were designed and the multi-unit ribozymes?in vitro transcription vector and retroviral vector were constructed. The in vitro cleavage ability was tested. The retroviral vector was transfected into K562 cell and the effects on proliferation, apoptosis, cell cycle and cell structure were observed. Results: Multi-unit ribozymes had in vitro cleavage efficiency of 70.8%, which was more efficient than single-unit and double-unit ribozymes. Transfection of the retroviral vector of the ribozyme into K562 cells, induced inhibition of cell growth and apoptosis. The incorporation rate of DNA in ribozymes transfected K562 cells was greatly decreased along with time passed, with an inhibition rate of more than 50% after 96 h of transfection. Under FCM, 18.4% of the cells underwent apoptosis 72 h after transfection and more cells were blocked in G phase, with the ratio in S phase greatly decreased (41.9%). Under electron microscope, compaction of nuclear chromatin and apoptosis bodies were observed. Conclusion: Multi-unit ribozymes specific to bcr-abl fusion gene can be used to treat CML and to purge bone marrow for self-grafting.

Cryptococcal Meningitis in Patient with Chronic Myeloid Leukemia

Objective: This study aimed to report the case of a female patient with chronic myeloid leukemia affected by cryptococcal meningitis. Case report: ML, white, 48 years old, female sex, previously diagnosed with chronic myeloid leukemia that has been refractive to the use of imatinib and who has recently begun using nilotinib, was admitted complaining of sudden and disabling migraine in the last 1 month associated with asthenia, adinamia, anorexia, disinterest for daily activities, dizziness, nausea, and vomiting. She evolved with ataxia, and started to stroll with help and showed decrease of muscular strength in her upper limbs. She also presented episodes of decrease of consciousness, with look fixation, no respond to sound stimulation, and short-term hearing loss. The cerebrospinal fluid showed presence of Cryptococcus sp. and, therefore, we began treatment with intravenous liposomal amphotericin B in the dose of 3 mg/kg/day, for 6 weeks. A new cerebrospinal fluid analysis, at the end of treatment, also showed rare structures that are compatible with Cryptococcus sp. As sequelae, she continued with hearing loss in her right ear and enhancement in her right auditory canal, seen in the magnetic resonance imaging. After stabilization and clinical improvement, she was discharged. After 3 weeks, she was hospitalized again with degeneration of the condition, and died due to intracranial hypertension secondary to cryptococcal infection. Final Considerations: This report reinforces the need of reflecting on fungi pathologies, especially in immunosuppressant patients, as well as the importance of early diagnosing and making a fast intervention, with the aims of providing quality of life and comfort to the patient and of minimizing neurological sequelae to the patient.