Acute myeloid leukemia (AML) can manifest as de novo AML (dn-AML) or secondary AML (s-AML), with s-AML being associated with inferior survival and distinct genomic characteristics. The underlying reasons for this disparity remain to be elucidated. In this multicenter study, next-generation sequencing (NGS) was employed to investigate the mutational landscape of AML in 721 patients from June 2020 to May 2023.Genetic mutations were observed in 93.34% of the individuals, with complex variations (more than three gene mutations) present in 63.10% of them. TET2, ASXL1, DNMT3A, TP53 and SRSF2 mutations showed a higher prevalence among older individuals, whereas WT1 and KIT mutations were more commonly observed in younger patients. BCOR, BCORL1, ZRSR2, ASXL1 and SRSF2 exhibited higher mutation frequencies in males. Additionally, ASXL1, NRAS, PPMID, SRSF2, TP53 and U2AF1 mutations were more common in patients with s-AML, which PPM1D was more frequently associated with therapy-related AML (t-AML). Advanced age and hyperleukocytosis independently served as adverse prognostic factors for both types of AML; however, s-AML patients demonstrated a greater number of monogenic adverse prognostic factors compared to dn-AML cases (ASXL1, PPM1D, TP53 and U2AF1 in s-AML vs. FLT3, TP53 and U2AF1 in dn-AML). Age and sex-related gene mutations suggest epigenetic changes may be key in AML pathogenesis. The worse prognosis of s-AML compared to dn-AML could be due to the older age of s-AML patients and more poor-prognosis gene mutations. These findings could improve AML diagnosis and treatment by identifying potential therapeutic targets and risk stratification biomarkers.

Background: Myeloid neoplasms (MN) tend to relapse and deteriorate. Exploring the genomic mutation landscape of MN using next-generation sequencing (NGS) is a great measure to clarify the mechanism of oncogenesis and progression of MN.Methods: This multicenter retrospective study investigated 303 patients with MN using NGS from 2019 to 2021. The characteristics of the mutation landscape in the MN subgroups and the clinical value of gene variants were analyzed.Results: At least one mutation was detected in 88.11% of the patients (267/303). TET2 was the most common mutation in the cohort, followed by GATA2, ASXL1, FLT3, DNMT3A, and TP53. Among patients with myeloid leukemia (ML), multivariate analysis showed that patients aged & GE;60 years had lower overall survival (OS, p = 0.004). Further analysis showed TET2, NPM1, SRSF2, and IDH1 gene mutations, and epigenetic genes (p < 0.050) presented significantly higher frequency in older patients. In patients with myelodysplastic syndrome (MDS) and myelodysplastic neoplasms (MPN), univariate analysis showed that BCORL1 had a significant impact on OS (p = 0.040); however, in multivariate analysis, there were no factors significantly associated with OS. Differential analysis of genetic mutations showed FLT3, TP53, MUC16, SRSF2, and KDM5A mutated more frequently (p < 0.050) in secondary acute myeloid leukemia (s-AML) than in MDS and MPN. TP53, U2AF1, SRSF2, and KDM5A were mutated more frequently (p < 0.050) in s-AML than in primary AML. KDM5A was observed to be restricted to patients with s-AML in this study, and only co-occurred with MUC16 and TP53 (2/2, 100%). Another mutation was MUC16, and its co-occurrence pattern differed between s-AML and AML. MUC16 mutations co-occurred with KDM5A and TP53 in 66.7% (2/3) of patients with s-AML and co-occurred with CEBPA in 100% (4/4) of patients with AML.Conclusions: Our results demonstrate different genomic mutation patterns in the MN subgroups and highlight the clinical value of genetic variants.

Regulation of cancer angiogenesis could be a useful strategy in cancer therapy. Metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) is a long non-coding RNA (lncRNA), and can induce cancer cell proliferation, while lncRNAs, generally are able to act as microRNA (miRNA) sponges. The latter is a type of competitive endogenous RNA (ceRNA) that regulates expression of the targeting miRNAs and protein-coding genes. This study investigated the proliferative role of MALAT1 in human umbilical vein endothelial cells (HUVECs) and the underlying molecular events. The data showed that knockdown of MALAT1 expression using MALAT1 siRNA inhibited HUVEC proliferation and also significantly decreased levels of FOXM1 mRNA and protein in vitro, while knockdown of FOXM1 expression reduced HUVEC proliferation. Annotation of HUVEC microarray data revealed that seven miRNAs, including miR-320a, were upregulated after knockdown of MALAT1 expression in HUVECs. MALAT1 was shown to reciprocally interact with miR-320a, i.e., expression of one negatively regulated levels of the other, whereas knockdown of MALAT1 expression promoted miR-320a levels. Furthermore, miR-320a could directly target and inhibit FOXM1 expression in HUVECs. Knockdown of MALAT1 expression enhanced miR-320a expression but reduced FOXM1 expression resulting in downregulation of HUVEC proliferation. However, such an effect was inhibited by miR-320a depletion. In conclusion, this study demonstrates that miR-320a plays an important role in mediating the effects of MALAT1 on HUVEC proliferation by suppression of FOXM1 expression. Thus, targeting of this gene pathway could be a novel strategy in cancer therapy.

Objective To observe the effect of electromagnetic radiation on the activation of JAK2 and STAT5 in the CD34 + cord blood stem cell.Methods CD34 + cord blood stem cells were isolated from the umbilical cord blood.Three exposed groups were administered with 1,5,25 mW/cm~2 electromagnetic exposure for 20 minutes,respectively.In the intervention group with the radiation power of 25 mW/cm~2,AG490 of the final concentration of 10~(-6) mol/L was pretreated for 30 minutes.The cell viabilitv (MTT),the rate of apoptosis (FCM),Caspase-3 protein level (WB) and the activation of JAK2 and STAT5 (WB) were assayed.Results Compared with the normal group,the cell viability of the electromagnetic group decreased at 12 h and 24 h,while apoptosis rate increased with dose.The activation of JA K2 and STAT5 increased at 12 h and 24 h and then decreased.The cell viability of the intervention group was significantly higher than the electromagnetic group with 25 mW/cm~2 irradiation at 12 h and 24 h,and apoptosis rate was significantly decreased as activation of JAK2 and STAT5 were lower. Conclusion JAK2 and STAT5 protein are involved in the process of the electromagnetic radiation within a scope of power (1-25 mW/cm~2) damaging CD34 + cord blood stem cells,suggesting that blocking the activation of JA K2 and STAT5 may be effective to alleviate the cell apoptosis of stem cells.

AIM: To construct a eukaryotic expression bicistron plasmid vector pIRES2-EGFP-FLT3 in order to investigate the role of FLt3 in the hemopoiesis and immune function during the recovery of acute radiation combined injury.METHODS: The experiment was completed in the Department of Occupational Hygiene of the Third Military Medical University of Chinese PLA between May and September 2003. After double digestion of pUMVC3-hFLT3 with SalⅠ and glⅡ, the product sequence (0.8 kb), was subcloned into the BglⅡsite of plasmid pIRES2-EGFP. After identification with NheⅠ and XhoⅠ digestion, the recombinant was transfected into CD34+ cells derived from human umbilical blood with liposome transfection technique, and screened with G418 followed by Flt3 fluorescence microscopy. RESULTS: ①A eukaryotic expression bicistron plasmid vector pIRES2-EGFP-FLT3 was constructed.②The recombined plasmid pIRES2-EGFP-FLT3 could be expressed in human umbilical blood CD34+ cells, the cDNA were confirmed by restriction enzyme digestion, sequencing. Green fluorescence was observed after its transfection into CD34+ cells. ③ Transfection efficiency was 10.3%.CONCLUSION: ① Plasmids vector pIRES2-EGFP- FLT3 with marker gene could be stably transfected and expressed in eukaryotic cells. ② The plasmid was transfected into isolated and purified CD34+ cells in umbilical blood, and the clone of positively transfected cells also can be obtained.

The immune system plays an important role in the treatment of chronic myeloid leukemia (CML). Identification of leukemia-associated antigens (LAAs) eliciting an immune response in patients is a prerequisite for specific immunotherapy of CML. To identify new LAAs in CML, We utilized a novel approach based serology and proteomics technologies. LAAs were identified by comparing the reactivity of proteins resolved by 2-DE with sera from CML patients and healthy donors. Several new LAAs were identified including alpha enolase, aldolase A, HSP70 protein8, beta-tubulin and tropomyosin isoforms. Although, the functions of these identified proteins in CML need further investigation, the detection of autoantibodies in CML may have value on CML screening, diagnosis, or follow-up. Additionally, identification of LAAs in CML may also be of vital importance in antigen-based immunotherapy.