Targeting UHRF1-SAP30-MXD4 axis for leukemia initiating cell eradication in myeloid leukemia.

Aberrant self-renewal of leukemia initiation cells (LICs) drives aggressive acute myeloid leukemia (AML). Here, we report that UHRF1, an epigenetic regulator that recruits DNMT1 to methylate DNA, is highly expressed in AML and predicts poor prognosis. UHRF1 is required for myeloid leukemogenesis by maintaining self-renewal of LICs. Mechanistically, UHRF1 directly interacts with Sin3A-associated protein 30 (SAP30) through two critical amino acids, G572 and F573 in its SRA domain, to repress gene expression. Depletion of UHRF1 or SAP30 derepresses an important target gene, MXD4, which encodes a MYC antagonist, and leads to suppression of leukemogenesis. Further knockdown of MXD4 can rescue the leukemogenesis by activating the MYC pathway. Lastly, we identified a UHRF1 inhibitor, UF146, and demonstrated its significant therapeutic efficacy in the myeloid leukemia PDX model. Taken together, our study reveals the mechanisms for altered epigenetic programs in AML and provides a promising targeted therapeutic strategy against AML.

Clinical Characteristics and Prognosis of 27 Patients with Childhood Acute Megakaryoblastic Leukemia.

BACKGROUND The aim of this study was to investigate the clinical features and prognostic factors of childhood acute megakaryoblastic leukemia (AMKL). MATERIAL AND METHODS The data of 27 cases of childhood AMKL admitted from November 2009 to July 2018 were retrospectively analyzed. The survival analysis and prognostic factors were analyzed by Kaplan-Meier method. RESULTS The median follow-up time was 26.4 months in 27 cases, and the complete response rate was 92.31% after 2 chemotherapy courses. Eight patients underwent bone marrow transplantation after 3-6 courses. Five patients died after transplantation, 4 of whom died due to recurrence after transplantation. Of the 27 patients, 10 developed recurrence (37.04%), and 8/10 had recurrence within 1 year. The 3-year overall survival rate and disease-free survival rates were (47±12)% and (36±14)%, respectively. Of the 27 AMKL cases, the 3 with Down syndrome (DS-AMKL) all survived after treatment, and the 3-year overall survival rate was 100%. However, of the other 24 AMKL patients without Down syndrome (non-DS-AMKL), 6 died and 6 abandoned treatment, and the 3-year overall survival rate was only 50%. Univariate analysis showed that 3-year overall survival rate was not correlated to gender, age, number of newly diagnosed white blood cells, karyotype, remission after 2 courses of treatment, and transplant after 3 courses of treatment of childhood AMKL cases. Nevertheless, recurrence and remission after 2 courses of treatment were significantly correlated with 3-year overall survival rate. CONCLUSIONS Children with non-DS-AMKL have a high degree of malignancy and are prone to early recurrence with a poor prognosis, whereas the prognosis of DS-AMKL is relatively good. Recurrence after treatment and remission after 2 courses of treatment are important factors influencing the prognosis of childhood AMKL. Recurrence after transplantation is the leading cause of death in transplantation patients.

SETD2 deficiency accelerates MDS-associated leukemogenesis via S100a9 in NHD13 mice and predicts poor prognosis in MDS.

SETD2, the histone H3 lysine 36 methyltransferase, previously identified by us, plays an important role in the pathogenesis of hematologic malignancies, but its role in myelodysplastic syndromes (MDSs) has been unclear. In this study, low expression of SETD2 correlated with shortened survival in patients with MDS, and the SETD2 levels in CD34+ bone marrow cells of those patients were increased by decitabine. We knocked out Setd2 in NUP98-HOXD13 (NHD13) transgenic mice, which phenocopies human MDS, and found that loss of Setd2 accelerated the transformation of MDS into acute myeloid leukemia (AML). Loss of Setd2 enhanced the ability of NHD13+ hematopoietic stem and progenitor cells (HSPCs) to self-renew, with increased symmetric self-renewal division and decreased differentiation and cell death. The growth of MDS-associated leukemia cells was inhibited though increasing the H3K36me3 level by using epigenetic modifying drugs. Furthermore, Setd2 deficiency upregulated hematopoietic stem cell signaling and downregulated myeloid differentiation pathways in the NHD13+ HSPCs. Our RNA-seq and chromatin immunoprecipitation-seq analysis indicated that S100a9, the S100 calcium-binding protein, is a target gene of Setd2 and that the addition of recombinant S100a9 weakens the effect of Setd2 deficiency in the NHD13+ HSPCs. In contrast, downregulation of S100a9 leads to decreases of its downstream targets, including Ikba and Jnk, which influence the self-renewal and differentiation of HSPCs. Therefore, our results demonstrated that SETD2 deficiency predicts poor prognosis in MDS and promotes the transformation of MDS into AML, which provides a potential therapeutic target for MDS-associated acute leukemia.

Different roles of E proteins in t(8;21) leukemia: E2-2 compromises the function of AETFC and negatively regulates leukemogenesis.

The AML1-ETO fusion protein, generated by the t(8;21) chromosomal translocation, is causally involved in nearly 20% of acute myeloid leukemia (AML) cases. In leukemic cells, AML1-ETO resides in and functions through a stable protein complex, AML1-ETO-containing transcription factor complex (AETFC), that contains multiple transcription (co)factors. Among these AETFC components, HEB and E2A, two members of the ubiquitously expressed E proteins, directly interact with AML1-ETO, confer new DNA-binding capacity to AETFC, and are essential for leukemogenesis. However, the third E protein, E2-2, is specifically silenced in AML1-ETO-expressing leukemic cells, suggesting E2-2 as a negative factor of leukemogenesis. Indeed, ectopic expression of E2-2 selectively inhibits the growth of AML1-ETO-expressing leukemic cells, and this inhibition requires the bHLH DNA-binding domain. RNA-seq and ChIP-seq analyses reveal that, despite some overlap, the three E proteins differentially regulate many target genes. In particular, studies show that E2-2 both redistributes AETFC to, and activates, some genes associated with dendritic cell differentiation and represses MYC target genes. In AML patients, the expression of E2-2 is relatively lower in the t(8;21) subtype, and an E2-2 target gene, THPO, is identified as a potential predictor of relapse. In a mouse model of human t(8;21) leukemia, E2-2 suppression accelerates leukemogenesis. Taken together, these results reveal that, in contrast to HEB and E2A, which facilitate AML1-ETO-mediated leukemogenesis, E2-2 compromises the function of AETFC and negatively regulates leukemogenesis. The three E proteins thus define a heterogeneity of AETFC, which improves our understanding of the precise mechanism of leukemogenesis and assists development of diagnostic/therapeutic strategies.

Clinical comparison of mifepristone and gestrinone for laparoscopic endometriosis.

Endometriosis is a common disease among women of childbearing age, and it is the main cause of dysmenorrhea and infertility. This article analyzes the efficacy of mifepristone and gestrinone in the treatment of endometriosis. The results showed that the recurrence rate of mifepristone group and gestrinone group were 8.33% and 5%, respectively, which was significantly lower than 23.33% of the control group. Before and after treatment, LH, endocrine test results FSH PRL had no obvious change in mifepristone group and gestrinone group, while E2 decreased, as mifepristone group (141.7±31.2) pmol/L, gestrinone group (64.2±11.7) pmol/L. The incidence of adverse reactions and liver dysfunction in the mifepristone group were significantly lower than those the gestrinone group (P<0.05). Mifepristone and gestrinone can be used for endometriosis postoperative adjuvant treatment, is safe and effective, but using mifepristone has the lower rate of adverse reaction. In conclusion, mifepristone is a current research focus, its mechanism of action in the process of exploration, has broad prospects in the treatment of endometriosis, its long-term application security is paid more and more attention.

[Lipid Composition of Different Breeds of Milk Fat Globules by Confocal Raman Microscopy].

Different breeds of cows affect the form of fat exist in dairy products and the final functionality, which depended mainly on the composition of the milk fat globules(MFG). However, the relationship between the composition and breeds has not been illuminated. In our study, differences in the lipid content and fatty acid composition of native bovine, buffalo and yak MFG were investigated by confocal Raman spectroscopy. The research offers the possibility of acquisition and analysis of the Raman signal without disruption of the structure of fat globule. The results showed that yak MFG had a higher ratio of band intensities at 2 885/2 850 cm(-1), indicating yak MFG tend to have a triglyceride core in a fluid state with a milk fat globule membrane in a crystalline state. The buffalo and yak MFG had a higher level of unsaturation compared to bovine MFG, shown by a higher ratio of band intensities at 1 655/1 744 cm(-1). The results indicate that small MFG of buffalo is more unsaturated than yak, while the large MFG of buffalo is less unsaturated than the yak. Thus, selective use of cream with yak MFG would allow a harder and more costly churning process but lead to a softer butter. Buffalo milk which contains larger MFG is more suitable for cream and MFG membrane separation.

Differential role of Id1 in MLL-AF9-driven leukemia based on cell of origin.

Inhibitor of DNA binding 1 (Id1) functions as an E protein inhibitor, and overexpression of Id1 is seen in acute myeloid leukemia (AML) patients. To define the effects of Id1 on leukemogenesis, we expressed MLL-AF9 in fetal liver (FL) cells or bone marrow (BM) cells isolated from wild-type, Id1(-/-), p21(-/-), or Id1(-/-)p21(-/-) mice, and transplanted them into syngeneic recipient mice. We found that although mice receiving MLL-AF9-transduced FL or BM cells develop AML, loss of Id1 significantly prolonged the median survival of mice receiving FL cells but accelerated leukemogenesis in recipients of BM cells. Deletion of Cdkn1a (p21), an Id1 target gene, can rescue the effect of Id1 loss in both models, suggesting that Cdkn1a is a critical target of Id1 in leukemogenesis. It has been suggested that the FL transplant model mimics human fetal-origin (infant) MLL fusion protein (FP)-driven leukemia, whereas the BM transplantation model resembles postnatal MLL leukemia; in fact, the analysis of clinical samples from patients with MLL-FP(+) leukemia showed that Id1 expression is elevated in the former and reduced in the latter type of MLL-FP(+) AML. Our findings suggest that Id1 could be a potential therapeutic target for infant MLL-AF9-driven leukemia.

The improvement effect and mechanism of citrus fiber on the water-binding ability of low-fat frankfurters.

Water-binding ability is a major quality attribute of low-fat meat products. This study aimed to evaluate the effects of citrus fiber on the water-binding ability of low-fat frankfurters. Low-fat (10% fat) frankfurters with different contents (0, 1, 2, and 3%) of citrus fiber were prepared. Their cooking loss, water distribution, microstructure, and protein structure were assessed and compared to those of normal-fat (20% fat) frankfurters. Results demonstrated that citrus fiber could significantly reduce the cooking loss of low-fat frankfurters from 12.69 to 4.71%. Adding citrus fiber led to a faster relaxation time in low-fat frankfurters, and a significant increase in the proportion of immobilized water. Frankfurters with citrus fiber exhibited a more compact, continuous protein network structure. Furthermore, the intensity changes in the Raman bands near 760 and 2930 cm-1 showed that citrus fiber increased hydrophobic interactions around hydrocarbon chains and tryptophan residues in myofibrillar proteins. In conclusion, citrus fiber effectively improved the water-binding ability of low-fat frankfurters by increasing the hydrophobic interactions of myofibrillar proteins and changing the frankfurter microstructure.

Eriocalyxin B ameliorates experimental autoimmune encephalomyelitis by suppressing Th1 and Th17 cells.

Eriocalyxin B (EriB), a diterpenoid isolated from Isodon eriocalyx, was previously reported to have antitumor effects via multiple pathways, and these pathways are related to immune responses. In this study, we demonstrated that EriB was efficacious in experimental autoimmune encephalomyelitis (EAE), an animal model for multiple sclerosis. Treatment with EriB led to amelioration of EAE, which correlated with reduced spinal cord inflammation and demyelination. EriB treatment abolished encephalitogenic T-cell responses to myelin oligodendrocyte glycoprotein in an adoptive transfer EAE model. The underlying mechanism of EriB-induced effects involved inhibition of T helper (Th) 1 and Th17 cell differentiation through Janus Kinase/Signal Transducer and Activator Of Transcription and Nuclear factor-κB signaling pathways as well as elevation of reactive oxygen species. These findings indicate that EriB exerts potent antiinflammatory effects through selective modulation of pathogenic Th1 and Th17 cells by targeting critical signaling pathways. The study provides insights into the role of EriB as a unique therapeutic agent for the treatment of autoimmune diseases.

Dissection of mechanisms of Chinese medicinal formula Realgar-Indigo naturalis as an effective treatment for promyelocytic leukemia.

To enhance therapeutic efficacy and reduce adverse effects, practitioners of traditional Chinese medicine (TCM) prescribe a combination of plant species/minerals, called formulae, based on clinical experience. Nearly 100,000 formulae have been recorded, but the working mechanisms of most remain unknown. In trying to address the possible beneficial effects of formulae with current biomedical approaches, we use Realgar-Indigo naturalis formula (RIF), which has been proven to be very effective in treating human acute promyelocytic leukemia (APL) as a model. The main components of RIF are realgar, Indigo naturalis, and Salvia miltiorrhiza, with tetraarsenic tetrasulfide (A), indirubin (I), and tanshinone IIA (T) as major active ingredients, respectively. Here, we report that the ATI combination yields synergy in the treatment of a murine APL model in vivo and in the induction of APL cell differentiation in vitro. ATI causes intensified ubiquitination/degradation of promyelocytic leukemia (PML)-retinoic acid receptor alpha (RARalpha) oncoprotein, stronger reprogramming of myeloid differentiation regulators, and enhanced G(1)/G(0) arrest in APL cells through hitting multiple targets compared with the effects of mono- or biagents. Furthermore, ATI intensifies the expression of Aquaglyceroporin 9 and facilitates the transportation of A into APL cells, which in turn enhances A-mediated PML-RARalpha degradation and therapeutic efficacy. Our data also indicate A as the principal component of the formula, whereas T and I serve as adjuvant ingredients. We therefore suggest that dissecting the mode of action of clinically effective formulae at the molecular, cellular, and organism levels may be a good strategy in exploring the value of traditional medicine.

[The investigation of JAK2 V617F point mutation in myeloproliferative disorders by allele-specific polymerase chain reaction in combination with sequence analysis].

OBJECTIVE: To study the JAK2 V617F point mutation in myeloproliferative disorders (MPD) and explore the clinical significance. METHODS: We used Allele-specific polymerase chain reaction (AS-PCR) in combination with sequence analysis to detect the mutation in genomic DNA of peripheral blood mononuclear cells from 20 chronic myelogenous leukemia (CML) patients, 23 polycythaemia vera (PV), 40 essential thrombocythaemia (ET), 8 idiopathic myelofibrosis (IMF), 3 hypereosinophilic syndrome (HES). RESULTS: JAK2 V617F was found in 38 (51.4%) of 74 BCR/ABL-negative MPD including 16 PV, 18 ET, 3 IMF and 1 HES patients. All positive samples and 10 negative samples identified by AS-PCR were confirmed by sequence analysis. Mutation-positive patients with ET had significantly increased hemoglobin, hematocrit, and neutrophil proportion than those without the mutation. CONCLUSION: JAK2 V617F mutation is the key molecular genetics feature of BCR/ABL-negative MPD. Detection of JAK2 V617F mutation will bring about a major impact to the diagnosis, classification and treatment of MPD.

[Application of a four antibody (cMPO/cCD79aalpha/cCD3/CD45) combination to the diagnosis of acute leukemia expressing cross-lineage antigens].

OBJECTIVE: To explore the diagnostic value of intracellular antibody combination in acute leukemia (AL) expressing cross-lineage cell-surface antigens. METHODS: Flow cytometric immunophenotyping using intracellular antibody combination (cMPO/cCD79alpha/cCD3/CD45) was performed additionally in 60 patients who expressed cross-lineage antigens from 269 previously untreated adult AL. RESULTS: Fifty-four of 269 previously untreated adult AL patients who expressed only one kind of intracellular antigen were diagnosed as cross-lineage AL, the percentage of cross-lineage AL in T cell acute lymphoblastic leukemia (T-ALL), B-ALL and acute myeloid leukemia (AML) was 28.6%, 43.6% and 13.4%, respectively. The positive rate of CD7, CD19, CD5 and CD20 in cross-lineage AML was 65.4%, 15.4%, 11.5%, and 7.7%, respectively. The positive rate of CD13, CD33 and CD15 in cross-lineage ALL was 89.3%, 21.4% and 3.6%, respectively. Six (2.3%) patients expressed two-lineage intracellular antigens were diagnosed as biphenotypic AL: 2 of T/B type and 4 B/M (B/myeloid) type. CONCLUSION: Intracellular antibodies possess lineage specificity and four-color combination flow cytometric immunophenotyping can provide fast and multi-parameter data. To ensure accuracy of the results, CD45/SSC gating and normal cells as internal reference should be used in the immunophenotyping of abnormal cells.

[Four-color flow cytometric immunophenotypic features of blasts in myelodysplastic syndromes].

This study was aimed at exploring the immunophenotypic features of blasts in patients with myelodysplastic syndromes (MDS). Four-color flow cytometry using conventional and secondary gating strategies was used to immunophenotype blasts and the CD34 positive cells in bone marrow nucleated cells of 29 patients with MDS. The results showed: (1) with progression of MDS from RA/RAS, RAEB to RAEB-T, the proportion of CD34(+) cells were gradually increased from 8.0%, 46.4% to 76.8% (P < 0.05); (2) using CD45 vs SSC gating strategy, with the transformation of RA/RAS, RAEB to RAEB-T, the expression of HLA-DR, CD13, CD33, CD117 were also gradually increased (P < 0.05), and the expression of CD15 was gradually decreased (P < 0.05); (3) using CD45 vs CD34 gating strategy, the expression of HLA-DR, CD13, CD33, CD117 on blasts were higher by secondary gating method than those by conventional gating (P < 0.05). However, there were no significant difference (P > 0.05) in the expression of above-mentioned antigens on CD34(+) cells among different MDS subtypes. It is concluded that conventional gating method can reflect MDS progression from RA/RAS, RAEB to RAEB-T, and secondary gating strategy may accurately reflect the biological features of blasts in MDS. Abnormal expression of CD34 is related to the immaturity level and heterogeneity of blast cells, which is beneficial to the diagnosis of clinically suspected MDS incapable of diagnosing with morphology and cytogenetics.

[ZAP-70 expression in 24 patients with B cell chronic lymphocytic leukemia].

To explore the ZAP-70 expression in chronic lymphocytic leukaemia (CLL) and its relationship with other prognostic factors, the expressions of ZAP-70 protein and CD38 in bone marrow or peripheral blood of 24 patients with B-CLL were determined by four-color flow cytometry. The results showed that ZAP-70 was positively expressed in 37.5% of all B-CLL patients, 20% (3/15) patients in Binet A stage, and 66.7% in Binet B + C. The expression of ZAP-70 had significant difference between Binet A and Binet B + C (P < 0.05). CD38 was expressed in 29.1% of B-CLL patients and 3 out of these cases were in stage A, 2 out of 3 cases in stage A co-expressed CD38 and ZAP-70. The expression of CD38 had no significant difference between Binet A and Binet B + C stage (P > 0.05). ZAP-70+ and CD38+ were both expressed in 83.3% of B-CLL patients (P < 0.05). It is concluded that ZAP-70 protein can be routinely measured by flow cytometry in the laboratory. High expression of ZAP-70 is correlated with other prognostic factors such as clinic stage, chromosome abnormalities and CD38 in B-CLL.