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.

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.

Acute Leukemia in Niger: Epidemiological, Diagnostic and Therapeutic Aspects

Objective: Improve the care of patients followed for acute leukemia in the Oncohematology department of the National Hospital of Niamey. Methods: This was a prospective study, over a period of 2 years from January 1, 2018 to December 31, 2019, in patients with acute leukemia in the Oncohematology department of the National Hospital of Niamey (HNN), whose diagnosis was made on a blood smear associated with a myelogram and immunophenotyping and who were consenting. Results: We collected 25 cases of acute leukemia confirmed by myelogram and immunophenotyping. The mean age of the patients was 31.32 years, with a predominance of women, a sex ratio of 0.92. Pupils and students were in the majority with 40% and most came from the Niamey region, i.e. 68%. Anemic syndrome was the most common clinical sign in 96%. ALL predominated in 64% of cases. On the blood count, the hyperleukocytosis was more marked in AML (mean white count: 197256.6 elts/mm3) than in ALL (137891.6 elts/mm3), it was the same for thrombocytopenia which is more marked in AML (75588.89/mm3) than in ALL (52156.25/mm3). Therapeutically, 52% of patients received chemotherapy. The mean overall survival was 16.223 ± 3.191 months, including a mean survival for AML of 6.853 ± 1200 months compared to 21.720 ± 5.920 months for ALL. Conclusion: Acute leukemia still remains a major problem in our context, due to the precariousness of limited financial, diagnostic and therapeutic resources. Thus reflecting in our results, the increasing number of cases, the diagnostic delay and the guarded prognosis. This is the reality in several other countries in the sub-region and even in certain developed countries.

Parametric and Non-Parametric Survival Analysis of Patients with Acute Myeloid Leukemia (AML)

Background: Acute Myeloid leukemia (AML) is the most prominent acute leukemia in adults. In the United States, we experience over 20,000 cases per year. Over the past decade, improvements in the diagnosis of subtypes of AML and advances in therapeutic approaches have improved the outlook for patients with AML. However, despite these advancements, the survival rate among patients who are less than 65 years of age is only 40 percent. Purpose: The purpose of the paper is to study if there exists any significant difference in the survival probabilities of male and female AML patients. Also, we want to investigate if there is any parametric probability distribution that best fits the male and female patient survival and compare the survival probabilities with the non-parametric Kaplan-Meier (KM) method. Methods: We used both parametric and non-parametric statistical methods to perform the survival analysis to assess the survival probabilities of 2015 patients diagnosed with AML. Results: We found evidence of a statistically significant difference between the mean survival time of male and female patients diagnosed with AML. We performed parametric survival analysis and found a Generalized Extreme Value (GEV) distribution best fitting the data of the survival time for male and female patients. We then estimated the survival probabilities and compared them with the frequently used non-parametric Kaplan-Meier (KM) survival method. Conclusion: The comparison between the survival probability estimates of the two methods revealed a better survival probability estimate by the parametric method than the Kaplan-Meier. We also compared the median survival time of male and female patients individually with descriptive, parametric, and non-parametric methods of analysis. The parametric survival analysis is more robust and efficient because it is based on a well-defined parametric probabilistic distribution, hence preferred over the non-parametric Kaplan-Meier estimate. This study offers therapeutic significance for further enhancement to treat patients with Acute Myeloid Leukemia.

Sunitinib Reduces Acute Myeloid Leukemia Clonogenic Cells in Vitro and Has Potent Inhibitory Effect on Sorted AML ALDH+ Cells

Sunitinib is an orally administered, multi-target tyrosine kinase inhibitor that has been approved by the FDA for the treatment of renal cell carcinoma and imatinib resistant gastro-intestinal tumors. Anti-leukemic activity of sunitinib has been examined in early clinical trials with limited success. However, recent trials on acute myeloid leukemia (AML) patients carrying FLT3 mutations have shown promising results. Effects of sunitinib on leukemic clonogenic cells and potential leukemic stem cells have not been examined so far. We analyzed the anti-proliferative and apoptotic properties of sunitinib on AML-derived cell lines. We also tested the effect of sunitinib on AML patient derived clonogenic cells (AML-CFC), as well as flow-sorted potential leukemic progenitors. Peripheral blood or bone marrow samples were obtained from newly diagnosed AML patients and flow sorted for CD34+ CD133+ or ALDH+ cells. Umbilical cord blood derived CD34+ cells were used as normal controls. Sunitinib induced growth arrest and apoptosis in AML derived cell lines. In addition, 7 μM sunitinib induced 75% reduction of AML-CFC as compared to DMSO treated control (±6.79%;n = 4). In contrast, 7 μM sunitinib treatment of umbilical cord blood derived normal CD34+ cells showed 29% reduction in AML-CFC (±6.77%;n = 5). Treatment of ALDH+ cells sorted from 2 AML cases and CD34+ CD133+ cells from one patient showed reduction of AML-CFC on treatment with sunitinib. Our study highlighted a potent anti-proliferative and proapoptotic effect of sunitinib on AML cell lines, AML patient derived clonogenic cells and potential leukemic stem cells.

Resveratrol-downregulated Phosphorylated Liver Kinase B1 Is Involved in Senescence of Acute Myeloid Leukemia Stem Cells

Summary： Senescence is an important obstacle to cancer development. Engaging a senescent response may be an effective way to cure acute myeloid leukemia （AML）. The aim of this study was to examine the effect of resveratrol-downregulated phosphorylated liver kinase B1 （pLKB1） on the senescence of acute myeloid leukemia （AML） stem cells. The protein expressions of pLKB 1 and Sirtuin 1 （SIRT1）, a regulator ofpLKB1, were measured in CD34＋CD38-KGla cells treated with resveratrol （40 μmol/L） or not by Western blotting. Senescence-related factors were examined, including p21 mRNA tested by real-time PCR, cell morphology by senescence-associated β-galactosidase （SA-β-gal） staining, cell pro- liferation by MTT assay and cell cycle by flow cytometry. Besides, apoptosis was flow cytometrically determined. The results showed that pLKB1 was highly expressed in CD34＋CD38- KGla cells, and resveratrol, which could downregulate pLKB1 through activation of SIRT1, induced senescence and apoptosis of CD34＋CD38- KGla cells. It was concluded that resveratrol-downregulated pLKB1 is in- volved in the senescence of AML stem cells.

Cutaneous Symptoms Revealing M5 Acute Myeloid Leukemia: A Case Report

Cutaneous presentation revealing acute myeloid leukemia (AML) is rare, and the prognosis is poor. We report a case of an 11-month-old male who presented with cutaneous lesions present as diffuse purplish papulo-nodular lesions, revealing M5 acute myeloid leukemia. Skin biopsy and immunohistochemical examination, combined with routine blood analysis and bone marrow examination, contributed to early diagnosis. However, despite intensive chemotherapy treatment, the prognosis was poor and death occurred during the first months of treatment.

Rare myeloid sarcoma/acute myeloid leukemia with adrenal mass after allogeneic mobilization peripheral blood stem cell transplantation

Myeloid sarcoma(MS)is a rare hematological neoplasm that develops either de novo or concurrently with acute myeloid leukemia(AML).This neoplasm can also be an initial manifestation of relapse in a previously treated AML that is in remission.A 44-year-old male patient was diagnosed with testis MS in a local hospital in August 2010.After one month,bone marrow biopsy and aspiration confirmed the diagnosis of AML.Allogeneic mobilization peripheral blood stem cell transplantation was performed,with the sister of the patient as donor,after complete remission(CR)was achieved by chemotherapy.Five months after treatment,an adrenal mass was detected by positron emission tomography-computed tomography(PET-CT).Radiotherapy was performed for the localized mass after a multidisciplinary team(MDT)discussion.The patient is still alive as of May 2013,with no evidence of recurrent MS or leukemia.

Analyses of therapeutic effects using rludarabine and cytarabine on acute myeloid leukemia at different stages during treatment

Objective: To evaluate the therapeutic effect of the fludarabine and cytarabine (FA) regimen on acute myeloid leukemia (AML) at different phases during treatment. Methods: A total of 185 patients with AML were divided into 4 groups based on the outcome of previous treatments. Patients in Group 1 had no remission after the first course of induction chemotherapy (n = 55). Patients in Group 2 had no remission after no less than two courses of induction chemotherapy (n = 41). Patients in Group 3 had early relapse (n = 40). Patients in Group 4 had late relapse (n = 49). Patients in groups 2, 3 and 4 had refractory AML or AML with relapse. We assessed the efficacy and toxicity of FA combination chemotherapy in each of these 4 groups. Results: The complete remission (CR) rates of Groups 1, 2, 3 and 4 were 74.5% (41/55), 45.9% (19/41), 17.5% (7/40) and 38.8% (19/49), respectively. The CR rate was higher in Group 1 than in the other 3 groups (34.6%, 45/130) (P = 0.000). A significant correlation was found between CR rate and the number of chemotherapeutic courses (P = 0.023). The main adverse reactions included bone marrow suppression and secondary infection. Conclusion: FA regimen is a good choice for patients with AML, especially those who have failed to achieve CR after the first course of induction chemotherapy.

Flow Cytometric Diagnosis of Acute Leukemia and Aberrant Antigen: Sohag University Experience

Background: Multiparameter flow cytometry is the most important method for the lineage assignment and maturational analysis of acute leukemias (AL) cells. The multi parametric immunophenotyping analysis allows the detection of aberrant antigen expression and the analysis of heterogeneity and clonality of malignant cells in AL. The aim of the work is to study the immunophenotypes of blasts from patients with AL and determine the frequency of aberrant markers. Subjects and Methods: Retrospective study to analyze immunophenotypic data of de novo 144 AL patients who were diagnosed in Clinical Pathology Department, Sohag University. Results: We found that 61.8% of AL patients were classified as acute myeloblastic leukemia (AML) while 38.9% classified as acute lymphoblastic leukemia (ALL). The commonest FAB subtype in AML group was AML-M2 (31.8%) followed by M4-M5 27.3%. As regard ALL, there were 85.7% with B-ALL and 14.3% with T-ALL. The aberrancy expressions were found in 66 of AL cases (45.8%), CD7 was the most commonly expressed lymphoid antigen in AML (25%), CD13 was the most commonly expressed myeloid antigen in ALL (39.3%). Conclusion: The multi parametric immunophenotyping analysis of AL is sufficient for diagnosis and classification of leukemia. The frequencies of

A Construction of Object Detection Model for Acute Myeloid Leukemia

The evolution of bone marrow morphology is necessary in Acute Mye-loid Leukemia(AML)prediction.It takes an enormous number of times to ana-lyze with the standardization and inter-observer variability.Here,we proposed a novel AML detection model using a Deep Convolutional Neural Network(D-CNN).The proposed Faster R-CNN(Faster Region-Based CNN)models are trained with Morphological Dataset.The proposed Faster R-CNN model is trained using the augmented dataset.For overcoming the Imbalanced Data problem,data augmentation techniques are imposed.The Faster R-CNN performance was com-pared with existing transfer learning techniques.The results show that the Faster R-CNN performance was significant than other techniques.The number of images in each class is different.For example,the Neutrophil(segmented)class consists of 8,486 images,and Lymphocyte(atypical)class consists of eleven images.The dataset is used to train the CNN for single-cell morphology classification.The proposed work implies the high-class performance server called Nvidia Tesla V100 GPU(Graphics processing unit).

Acute Leukemia in Children

Acute leukemia is the most common childhood cancer and accounts for 31% of all cancers in children. There are two main types of acute leukemia. The most common is ALL （acute lymphoblastic leukemia） affecting the lymphoid lineage, and the more rare AML （acute myeloid leukemia） affecting the myeloid linage. The intention of this thesis is to follow the course of treatment from the admission to the hospital until the last check up and also see how a child will react to the treatment and side effects in later life. We studied literature and my own case records from the period when I was treated for ALL. From the literature and my case records, we can see that children tolerate treatment quite well. Due to rapid diagnostics and the possibility to give high doses chemotherapy, the overall prognosis appears to be very good. Today, acute leukemias of paediatric patients have a really favourable prognosis. The overall survival rate for ALL is higher than 80% and for AML 65%. So the results are good, but there is still a long way to go before we can be satisfied. To date we do not have a contingency program for children treated for acute leukemia after 18 years of age （neither in Norway or Slovakia） so perhaps this should be a focus point in the future. It could be extended to follow up patients in adulthood in order to monitor late effects that may occur in later life after many years of treatment.

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.

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

地西他滨联合改良CAG方案治疗AML1-ETO阳性复发、难治急性髓系白血病的临床研究

本研究旨在分析AML1-ETO阳性复发、难治急性髓系白血病的临床特点及地西他滨联合改良CAG方案对该类白血病患者的治疗效果及副作用。回顾性分析2013年1月至8月5例AML1-ETO阳性复发、难治急性髓系白血病,并分析临床特点,包括年龄、性别、初诊时伴随症状、外周血和骨髓特点等;同时分析地西他滨联合改良的CAG方案对该类患者的治疗效果和副作用。5例患者中复发2例,难治合并复发3例,初治时白细胞中位数12.55(7.8-66.55)×109/L、血小板中位数44(20-72)×109/L,血红蛋白中位数110(77-128)g/L、乳酸脱氢酶中位数312.9(123.6-877.8)μ/L。治疗结果表明:经过1个疗程地西他滨联合改良的CAG方案治疗后4例完全缓解,1例未缓解,总缓解率为80%;治疗的副作用主要为骨髓抑制,1例未缓解经FLAG方案治疗后在移植动员过程中突发心衰死亡。结论:初步结果表明AML1-ETO阳性复发、难治急性髓系白血病经本方案治疗后,其缓解率比较高、并发症较少,值得进一步研究和在临床推广应用。

急性白血病Ly+AML型和My+ALL型预后因素的临床研究

为了研究Ly+AML(表达淋巴系抗原的急性髓性白血病),My+ALL(表达髓系抗原的急性淋巴细胞白血病),AML(急性髓性白血病),ALL(急性淋巴细胞白血病)和BAL(急性双表型白血病)的预后,采用CD45/SSC双参数散点图设门,应用三色流式细胞术,对197例AL(急性白血病)初诊患者骨髓标本进行免疫分型,采用EGIL(白血病免疫分型欧洲协作组)积分系统,将患者分为5组:ALL43例,AML53例,Ly+AML39例,My+ALL53例,BAL9例。结果表明:Ly+AML(淋系抗原以CD7表达最常见,占53.8%)与My+ALL(髓系抗原以CD13表达最常见,占47.2%)相比,在白细胞数>100×109/L的例数、CD34阳性率及完全缓解(CR)率方面,差别无统计学意义(P>0.05),但在肝、脾、淋巴结肿大的例数方面,差别有统计学意义(P<0.05),My+ALL的例数相对更多。ALL与My+ALL在白细胞数>100×109/L的例数、肝脾淋巴结肿大的例数、CD34阳性率及CR率方面,差别均无统计学意义(P>0.05)。AML与Ly+AML相比,在白细胞数>100×109/L的例数、肝脾淋巴结肿大的例数及CD34阳性率方面,差别无统计学意义(P>0.05),但在CR率方面,差别有统计学意义(P<0.05),AML患者的CR率相对更高。BAL与Ly+AML和My+ALL相比,虽然BAL患者的CR率(仅37.5%)明显低于前两者(分别低了16.8%和27.8%),但是由于BAL的例数太少,差别并无统计学意义(P>0.05)。结论:Ly+AML的临床化疗可能应兼顾AML+ALL的两方面,因为它的预后因淋系抗原的表达而更差;而对于My+ALL来说,它的预后并没有因髓系抗原的表达而与ALL表现出明显差异,因此可以考虑采用与ALL相同的化疗方案。

AML-M1相关TCR Vβ克隆性增殖T细胞的研究

目的 了解急性髓细胞白血病M1亚型 (AML M1 )患者外周血TCRVβ亚家族T细胞的克隆性分布特点。 方法采用RT PCR扩增 9例M1患者外周血的单个核细胞的TCRVβ2 4个亚家族基因 ,分析其TCRVβ亚家族的利用情况。PCR产物进一步经基因扫描分析 ,了解其CDR3长度以判断T细胞的克隆性。结果 不同标本中可检测到 2～ 5个Vβ亚家族T细胞 ,以Vβ2、Vβ3、Vβ1 7和Vβ1 9为常见 ,并在Vβ2、Vβ3、Vβ5、Vβ8、Vβ9和Vβ1 9中发现克隆性生长T细胞。结论 M1病人外周血TCRVβ亚家族T细胞表达呈现明显的选择性和克隆性增殖情况 ,这可能与M1细胞相关抗原有关 ,且克隆性增殖Vβ亚家族分布以个体特异性为主。

AML-1/ETO基因相关的AML-M_2型白血病免疫表型分析

本研究探讨AML/ETO基因重排的FAB分型为AML-M2患者的骨髓免疫表型的特征和预测性,以及与急性早幼粒白血病(APL)的区别。应用流式细胞术分析17例经荧光原位杂交(FISH)检测AML-1/ETO融合基因阳性、FAB分型为AML-M2(M2/ETO+)患者骨髓的免疫表型,并与34例AML-1/ETO阴性、FAB分型AML-M3、临床诊断为APL患者进行了比较。结果发现,17例M2/ETO+患者骨髓中均可见一群(15.89%-68.53%)原始细胞和一群SSC较高异质性的粒细胞。原始细胞表达干细胞相关抗原CD34、HLA-DR和髓系抗原CD33、CD13、MPO。在M2/ETO+患者中CD33表达强度显著低于APL患者(P<0.001),HLA-DR、CD19和CD34+CD56+共表达的阳性率均显著高于APL患者(P<0.001),而CD9的表达率则显著低于APL患者(P<0.001)。M2/ETO+患者粒细胞表达分化的髓系抗原CD11b和CD15,并可分为CD15+CD11b-和CD15+CD11b+两群,而成熟的粒细胞标志CD10均为阴性,与早幼粒细胞的表达图形相似。17例(100%)M2/ETO+患者的粒细胞均表达CD56,显著高于M3患者(6/34例,17.56%)。结论:AML-1/ETO基因重排的AML-M2型(FAB分型)白血病具有独特的免疫表型,对其基因重排有较高的预测性。应用多参数免疫分型技术可较容易地将M2/ETO+亚型白血病与APL鉴别。

地西他滨联合CAG方案治疗AML1-ETO^+ AML的临床效果

目的:观察AML1-ETO^+复发/难治急性髓系白血病患者经地西他滨联合CAG方案治疗的临床治疗效果。方法:收集2015年6月至2016年1月我院收治的8例AML1-ETO+复发/难治急性髓系白血病患者临床资料,分析患者入院一般资料及初诊伴随症状、骨髓特点等,同时分析患者经地西他滨联合CAG方案治疗的临床观察指标及不良反应。结果:8例患者中复发1例,难治合并复发7例,初治时白细胞23.57(7.5-65.29)×10~9/L、血小板40(19-69)×10~9/L,血红蛋白107(79-131)g/L、乳酸脱氢酶313.5(124.1-865.9)U/L。8例患者经地西他滨联合CAG方案1个疗程治疗后7例症状完全缓解,1例症状未缓解,总缓解率为87.5%;本方案的不良反应为骨髓抑制,1例症状未缓解患者经FLAG方案治疗后突发心衰死亡。结论:AML1-ETO^+复发/难治急性髓系白血病患者经地西他滨联合CAG方案治疗后,各临床观察指标较理想,缓解率较高,并发症较少。

CD56表达对伴有AML/ETO的急性髓系白血病患者预后影响的分析

目的:探讨CD56表达对伴有AML/ETO的急性髓系白血病患者的预后影响。方法:回顾性分析西安交通大学第一附属医院2012年1月至2016年12月60例伴有AML/ETO的初发AML患者临床资料,比较CD56^+阳性与阴性组的临床特征。结果:CD56^+与CD56^-2组患者在1个疗程诱导缓解治疗后CR率、1年复发率和基因突变率方面有明显的差异(P<0.05);CD56^+患者RFS和OS均低于CD56^-患者,差异有统计学意义(P<0.05)。结论:CD56^+可能是伴有AML/ETO的AML患者预后差的重要因素。