Identification of cystic fibrosis transmembrane conductance regulator as a prognostic marker for juvenile myelomonocytic leukemia via the whole-genome bisulfite sequencing of monozygotic twins and data mining.

Background: Linked deoxyribonucleic acid (DNA) hypermethylation investigations of promoter methylation levels of candidate genes may help to increase the progressiveness and mortality rates of juvenile myelomonocytic leukemia (JMML), which is a unique myelodysplastic/myeloproliferative neoplasm caused by excessive monocyte and granulocyte proliferation in infancy/early childhood. However, the roles of hypermethylation in this malignant disease are uncertain. Methods: Bone marrow samples from a JMML patient and peripheral blood samples from a healthy monozygotic twin and an unrelated healthy donor were collected with the informed consent of the participant's parents. Whole-genome bisulfite sequencing (WGBS) was then performed. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed to analyze specific differentially methylated region (DMG) related genes. The target genes were screened with Cytoscape and Search Tool for the Retrieval of Interacting Genes/Proteins (STRING), which are gene/protein interaction databases. A data mining platform was used to examine the expression level data of the healthy control and JMML patient tissues in Gene Expression Omnibus data sets, and a survival analysis was performed for all the JMML patients. Results: The STRING analysis revealed that the red node [i.e., the cystic fibrosis transmembrane conductance regulator (CFTR)] was the gene of interest. The gene-expression microarray data set analysis suggested that the CFTR expression levels did not differ significantly between the JMML patients and healthy controls (P=0.81). A statistically significant difference was observed in the CFTR promoter methylation level but not in the CFTR gene body methylation level. The overall survival analysis demonstrated that a high level of CFTR expression was associated with a worse survival rate in patients with JMML (P=0.039). Conclusions: CFTR promoter hypermethylation may be a novel biomarker for the diagnosis, monitoring of disease progression, and prognosis of JMML.

TSST-1 protein exerts indirect effect on platelet activation and apoptosis.

Thrombocytopenia or platelet dysfunction is a risk factor for severe infection. Staphylococcus aureus (S. aureus) releases a variety of virulence factors especially toxic shock syndrome toxin 1 (TSST-1), which may cause toxic shock syndrome. S. aureus, when carrying the tst gene, is more prone to cause toxic shock syndrome and is responsible for an especially high rate of mortality. However, the effect of TSST-1 protein on platelets is unknown. Patients with the tst gene positive S. aureus bacteremia showed more serious infection, higher mortality and lower platelet count. The tst gene positive S. aureus strains induce more platelet apoptosis and activation and corresponding up-regulation of Bak and down-regulation of Bcl-XL in addition to the activation of Caspase-3. C57BL/6 mice infected with the tst gene positive strains resulted in both a decrease in platelet count and an increase in platelet apoptosis and/or activation events and mortality. Moreover, TSST-1 protein, encoded by tst gene, caused the decrease of platelet count, the increase of platelet apoptosis and activation events and the level of inflammatory cytokines in vivo. However, TSST-1 protein was unable to induce traditional activation and apoptosis on human platelets in vitro. These results suggested that TSST-1 protein may exert indirect effects on platelet activation and apoptosis in vivo.

[Prognostic significance and risk factors of minimal residual disease ≥1% on 19th day of induction chemotherapy in children with acute lymphoblastic leukemia].

OBJECTIVE: To assess the prognostic value of minimal residual disease on 19th day of induction chemotherapy (D19 MRD) and the risk factors of D19 MRD ≥ 1% in children with acute lymphoblastic leukemia (ALL) treated following the Chinese Children's Cancer Group ALL protocol. METHODS: We retrospectively analyzed the data of 243 children with ALL diagnosed between January 1, 2015 and December 31, 2018 in the Department of Pediatrics of Nanfang Hospital (Guangzhou China). Kaplan Meier-survival analysis was performed to compare the survival time between the patients with D19 MRD < 1% and those with D19 MRD ≥ 1%; logistic regression analyisis and Chi-square test were used to identify the risk factors of D19 MRD ≥ 1%. RESULTS: Compared with those with D19 MRD ≥ 1%, the children with D19 MRD < 1% had significantly better 3-year overall survival (100% vs 90.2%, P=0.004) and event-free survival (97.6% vs 71.6%, P < 0.001). Univariate analysis showed that the odds ratio (OR) for mediastinal invasion, T-cell immunophenotype, TEL/AML1 fusion gene and the presence of blasts in peripheral blood on the 5th day were 4.47 (95%CI: 0.275-72.968, P=0.034), 5.250 (95%CI: 1.950-14.133, P=0.02), 0.330 (95%CI: 0.112-0.970, P=0.036) and 4.407 (95%CI: 1.782-10.895, P=0.01), respectively. The initial risk stratification (P < 0.001), white blood cell grades (P=0.018) and its counts (P=0.027), and the number of blasts on the 5th day (P < 0.001) were significantly different between the two groups. Multivariate analysis showed that initial risk stratification as intermediate and high risks (OR=2.889, 95% CI: 1.193-6.996) and the presence of blasts in peripheral blood on the 5th day (OR=4.477, 95% CI: 1.692-11.843) were independent risk factors for poor early treatment response. CONCLUSIONS: D19 MRD ≥ 1% is a predictor of poor prognosis in children with ALL. Mediastinal invasion, T-cell immunophenotype and the presence of blasts in peripheral blood on the 5th day are all risk factors for poor early treatment response, while TEL/AML1 fusion gene is a protective factor; the initial risk stratification as intermediate to high risk and the presence of blasts in peripheral blood on the 5th day are independent risk factors for poor early treatment response of the patients.

[Effect of cyclophosphamide on hematopoietic stem cells in mice with iron overload].

OBJECTIVE: To explore the effect of cyclophosphamide on hematopoietic stem cells (HSCs) in mice with iron overload. METHODS: Mouse models of iron overload were established in 30 male C57BL/6 mice by intraperitoneal injections of iron dextran at low (0.25 g/kg), moderate (0.5 g/kg), and high (1 g/kg) doses (n=10), with another 10 PBS-treated mice as the control group. The changes in body weight, liver, spleen and bone marrow of the mice were recorded, and serum level of ferritin was detected. The mice receiving a moderate dose of iron dextran were further divided into 8 groups for observation at different time points (D1, D2, D3, D4, D5, D6, D7, and D14 groups) and were given intraperitoneal injection of 50 mg/kg cyclophosphamide (Cy) for 2 consecutive days. Peripheral blood cells, bone marrow mononuclear cells (BMMNCs), and the frequencies of different HSCs (HPCs, HSCs, LT-HSCs) in the BMMNCs were monitored. The cell cycle distribution in the HSCs, level of reactive oxygen species and the microenvironment of the HSCs were analyzed using flow cytometry. RESULTS: Compared with the control mice, the mice with iron overload showed obvious weight loss with significantly increased serum ferritin level, enlargement of the liver and spleen, and iron deposition in the organs (P < 0.05). No significant changes were noted in the peripheral blood of the mice with iron overload. The cyclophosphamide-treated mice exhibited significantly decreased number of WBCs and lymphocyte ratio at days 1 to 4 (P < 0.05). The numbers of BMMNCs and HPCs in mice with iron overload did not show significant changes as compared with those in the control mice, but the numbers of HSCs and LTHSCs decreased significantly in the mice with iron overload (P < 0.05). In cyclophosphamide-treated mice, the number of HSCs increased since day 1 and reached the peak level on day 3 (P < 0.05). Compared with those in the control group, the HSCs did not exhibit significant changes in cell cycle distribution in mice with iron overload, but the proportion of G0/G1 cells decreased significantly in cyclophosphamide group since day 1 and reached the lowest level on day 3 (P < 0.05). CONCLUSIONS: Iron deposition in the bone marrow causes long- term damages of the HSCs in the bone marrow but does not induce obvious changes in the peripheral blood. In mice with iron overload, intraperitoneal injection of 50 mg/kg cyclophosphamide for two days promotes cell cycle changes of the resting HSCs to mobilize the HSCs, and this effect is the most obvious on day 4.

Increased risk of arterial thromboembolic events in transfusion-naïve thalassemia: a nationwide population-based study.

Transfusion-naïve thalassemia minor/trait is often associated with decreased risk of coronary artery diseases. The present study aimed to evaluate the effect of transfusion-naïve thalassemia on the incidence of arterial thromboembolic events using the National Health Insurance Research Database, Taiwan (2001-2010). Data from patients with transfusion-naïve thalassemia (n=2356) frequency matched with non-thalassemia subjects (n=9424) according to sex, age, and index year at a ratio of 1:4 were included. The risk of arterial thromboembolic events, cerebrovascular ischemic events, arterial embolism/thrombosis, peripheral embolism, myocardial infarction, myocardial ischemia, and angina pectoris in transfusion-naïve thalassemia were analyzed using Cox proportional hazard regression models. The transfusion-naïve thalassemia group had significantly higher risk of arterial thromboembolic events (aHR=1.28, 95% CI 1.07 to 1.52) and myocardial ischemia (aHR=1.41, 95% CI 1.13 to 1.76) as compared with the non-thalassemia group. In addition, they also had a significantly higher cumulative incidence of arterial thromboembolic event and myocardial ischemia. Interestingly, a higher risk of arterial thromboembolic events (aHR=1.58, 95% CI 1.22 to 2.04) and myocardial ischemia (aHR=1.73, 95% CI 1.25 to 2.41) was observed in men with thalassemia as compared with those without. Furthermore, patients with comorbidities had an increased risk of arterial thromboembolic events than did those without comorbidities. The effect of thalassemia on arterial thromboembolic events may be mainly attributed to the influence of thalassemia on myocardial ischemia, as no significant differences were observed in other outcomes evaluated in the present study. In conclusion, the present study confirms the increased risk of arterial thromboembolic events, mainly attributed to the dramatic increase in myocardial ischemia, inminor patients with transfusion-naïve thalassemia.

Fabrication of multifunctional monometallic nanohybrids for reactive oxygen species-mediated cell apoptosis and enhanced fluorescence cell imaging.

Novel fluorescent monometallic Ag nanohybrids (Ag NHs) have been synthesized via polycytosine-mediated biomineralization on Ag nanoparticles (Ag NPs). Thiolated polycytosine dC12 was anchored on the Ag NPs, followed by dC12-templated synthesis of the Ag nanoclusters (Ag NCs). The Ag NHs exhibited strong fluorescence emission and improved resistance to dissolved oxygen in solution as compared to the Ag NCs. As an efficient therapeutic agent, the delivery of aptamer-functionalized Ag NHs into target cells induced reactive oxygen species (ROS) generation and cell apoptosis. The ROS-mediated apoptotic effect of the Ag NHs was stronger than that of the Ag NCs. For cell imaging, the target cells treated with aptamer-functionalized Ag NHs exhibited a higher fluorescence brightness than those treated with Ag NCs. Due to the fluorescence quenching of Ag NHs by the intracellularly generated ROS, the Ag NHs served as a fluorescent indicator for real-time monitoring of ROS-mediated cell apoptosis. The In vivo antitumor efficacy of the Ag NHs was also evaluated.