Philadelphia-negative myeloproliferative neoplasms are clonal blood disorders characterized by abnormal blood cell production. This study explores the clinical and epidemiological profiles of 111 Ecuadorian patients diagnosed with Philadelphia-negative myeloproliferative neoplasms, including polycythemia vera, essential thrombocythemia, and primary myelofibrosis, between 2014 and 2023. Patients were treated in different institutions, with clinical data collected on disease progression, complications, and survival. Polycythemia vera was the most common subtype (45.9%), followed by essential thrombocythemia (42.3%) and primary myelofibrosis (9%). The JAK2 V617F mutation was most prevalent in essential thrombocythemia (53.2%) and polycythemia vera (41.2%). Hydroxyurea, the most widely used treatment, was prescribed to 77% of the patients. Disease progression to myelofibrosis occurred in three polycythemia vera and two essential thrombocythemia cases, meanwhile One case of primary myelofibrosis and one case of myeloproliferative neoplasm, unclassified, progressed to acute myeloid leukemia. Survival rates varied across the cohort; notably, certain patients with polycythemia vera and essential thrombocythemia achieved survival durations of up to 19 years. These results reveal a relatively homogeneous epidemiological profile across the Latin American region and underscore the need for more multicenter studies to better characterize pH- MPNs in Ecuador and the region, to optimize diagnostic and treatment strategies.

Alkaline phosphatase (ALP) enzymatic activity has been proposed as a marker for distinguishing canine acute leukaemia (AL) subtypes (i.e., myeloid vs. lymphoid). However, ALP enzymatic activity has not been fully evaluated in CD34+ AL. Determine whether ALP enzymatic activity can differentiate CD34+ AL subtypes in dogs and distinguish CD34+ AL from CD34- haematopoietic tumours in tissue/effusion samples. Peripheral blood from 64 dogs with CD34+ AL, 10 with B cell chronic lymphocytic leukaemia (CLL), and 10 healthy controls were prospectively evaluated for ALP enzymatic activity via cytochemical staining; a subset also underwent ALP detection by flow cytometry (FC). Archived cytology slides from 67 tissue/effusion specimens, including 27 CD34+ AL, 22 T cell lymphomas, and 18 B cell lymphomas, were retrospectively assessed. CD34+ AL cases were categorised as acute myeloid leukaemia (AML), acute lymphoid leukaemia (ALL) or acute unclassifiable leukaemia (AUL) by established FC criteria. ALP positivity was defined as > 3% ALP+ neoplastic cells, which was selected based on receiver operating characteristic (ROC) curve analysis. Cytochemical ALP activity was detected in 61/64 (95.3%) CD34+ AL cases, with no significant differences between AML, ALL, and AUL subtypes (p > 0.05). All lymphoma and B cell CLL cases were ALP-negative. FC-based ALP analysis showed poor concordance with cytochemistry, and the correlation between %CD34 + ALP+ cells and %ALP+ neoplastic cells was weak (Spearman's ρ = 0.25). While ALP enzymatic activity is present in most CD34+ AL cases, it does not reliably differentiate CD34+ AL subtypes via cytochemistry. However, ALP may help distinguish CD34+ AL from B and T cell lymphomas. FC-based ALP analysis is not a reliable marker for CD34+ AL classification.

We present a case of acute clonal bone marrow 98% infiltration of atypical myeloid cells with borderline hypogranular/agranular promyelocytes/myelocytes and occasional blast cells maturity, which also formed extramedullary tumours in the chest wall, with isolated trisomy of chromosome 6 and pathogenic variant U2AF1 (S34F) that escapes established acute myeloid leukaemia (AML) diagnostic criteria according to the World Health Organization (WHO) classification. Following standard daunorubicin and cytarabine induction therapy, the disease progressed with the appearance of a previously undetected clone of leukaemic cells with a distinct immunophenotype demonstrating monocytoid differentiation and clonal evolution to a hypo-tetraploid karyotype with an average number of 84 chromosomes and new pathogenic NRAS and ZRSR2 mutations. The patient reactivated refractory disseminated intravascular coagulation (DIC) leading to a progressive supratentorial hematoma and finally cardiac arrest. In conclusion, our report shows that atypical clonal myelocytes can massively infiltrate the bone marrow and form extramedullary tumours, justifying the diagnosis and treatment of acute leukaemia, although they did not fit the current classification.

To investigate the clinical application value of artificial intelligence (AI)-based bone marrow cell recognition and analysis system in the diagnosis of hematological diseases. The bone marrow smears of hematological patients who were admitted to The Second Hospital of Shanxi Medical University from 2018 to 2020 were retrospectively analyzed. A total of 115 bone marrow smears with clear diagnosis and typical cell morphology characteristics were selected, including 20 cases of immune thrombocytopenia(ITP), 11 cases of iron deficiency anemia (IDA), 17 cases of megaloblastic anemia (MA), 20 cases of chronic myeloid leukemia (CML), 17 cases of acute lymphoblastic leukemia (ALL), 23 cases of acute promyelocytic leukemia (APL), and 7 cases of acute myeloid leukemia unclassified (AML-M2). The samples were analyzed by manual microscopic examination, AI automatic recognition, and manual correction after AI recognition. The images captured by the AI device were clear, and the cell morphological structures were distinct. The average experimental diagnostic efficiency parameters of the bone marrow nucleated cells classified in this system were calculated. The sensitivity was 74.90%, specificity was 99.03%, and accuracy was 98.29%. In the comparison between the AI recognition group and the manual examination group, the data of IDA, ITP, MA, and CML diseases were all greater than 0.85 in ICC correlation coefficient, with excellent consistency; the data of APL, AML-M2, and ALL three diseases were between 0.6 and 0.85 in ICC correlation coefficient, with moderate consistency. However, after manual review and correction, the ICC correlation coefficient between the data of the AI correction group and the data from the manual examination group was greatly improved. The AI bone marrow cell recognition and analysis system has the characteristics of high accuracy, high specificity, good sensitivity and fast detection. When used in combination with manual review, it can improve the detection efficiency of bone marrow cells morphological analysis and meet the needs of clinical work. 人工智能骨髓细胞识别分析系统在血液病辅助诊断中的应用. 探讨基于人工智能（Artificial Intelligence，AI）的骨髓细胞识别分析系统在临床血液系统疾病诊断工作中的应用价值。. 回顾性分析2018年至2020年于山西医科大学第二医院就诊的血液病患者的骨髓涂片,选取已明确诊断且细胞形态特征较为典型的骨髓涂片样本115例，其中免疫性血小板减少性紫癜（ITP)20例，缺铁性贫血(IDA)11例，巨幼细胞贫血(MA)17例，慢性髓系白血病（CML）20例，急性淋巴细胞白血病(ALL)17例，急性早幼粒细胞白血病(APL)23例，急性髓系白血病未分化型（AML-M2）7例，通过人工镜检、AI自动识别、AI识别后人工校正三种方法对样本进行细胞计数和形态学分析。. AI骨髓细胞识别分析系统所采集的图像清晰，细胞的形态结构清楚。AI自动识别分析常见骨髓细胞的准确度平均值为98.29%、敏感度平均值为74.90%、特异度平均值为99.03%。AI识别组与人工镜检组数据比较中，IDA、ITP、MA、CML类疾病的数据与人工镜检组的数据间ICC相关系数均大于0.85，一致性极好；APL、AML-M2、ALL三类疾病人工识别组数据与人工镜检组的数据间ICC相关系数在0.6-0.85之间，一致性一般；但经过人工审核校正后的AI校正组数据与人工镜检组数据间ICC相关系数得到很大提升。. AI骨髓细胞识别分析系统具有准确度、特异度高，敏感度好，检测快等特点，与人工审核相结合使用，可以提高骨髓细胞形态分析检测效率，满足临床工作需求。.

Homoharringtonine (HHT), a natural product used clinically to treat acute myeloid leukemia (AML) in combination with chemotherapy, induced transient remission in 25 % of unclassified AML patients when administered alone. Recent studies showed that HHT could be combined with targeting agents for AML treatment. However, the rationales of these combinations require further exploration to optimize therapeutic efficacy. To explore the sensitive subgroup of AML cells to HHT and to evaluate its optimal combinations with FLT3-ITD or Bcl-2 inhibitors based on apoptosis induction. AML cell lines harboring FLT3-ITD mutation and wild-type FLT3 were used to compare the apoptosis induction of HHT alone and in combination with venetoclax or FLT3-ITD inhibitor. siRNA and Western blotting were used to identify the key factors in the apoptosis induction. Xenograft models were used to test the in vivo antileukemia efficacy and toxicity. FLT3-ITD AML cell lines and primary AML cells exhibited greater sensitivity to HHT-induced apoptosis and in vivo antileukemia effects than FLT3 wild-type cells. HHT induces apoptosis through repressing the antiapoptotic protein Mcl-1 and activating the proapoptotic protein Bax, which is attenuated by Mcl-1 overexpression and Bax knockdown. HHT in combination with Bcl-2 inhibitor venetoclax induce synergistic apoptosis across all cell lines with enhanced toxicity observed in vivo. HHT plus FLT3 inhibitors, including midostaurin, quizartinib, gilteritinib and sorafenib, induce selective synergistic apoptosis in FLT3-ITD AML cells, with the most potent effect observed in the combination of HHT and quizartinib. HHT plus quizartinib significantly prolonged FLT3-ITD AML xenograft survival without causing toxicity. FLT3-ITD AML cells represent a responsive subgroup to HHT treatment in vitro and in vivo. The combination of HHT and quizartinib demonstrates optimal efficacy and selectivity against FLT3-ITD AML cells in xenografts, with no observed toxicity.