Effect of physical activity on anxiety, depression and obesity index in children and adolescents with obesity: A meta-analysis.

FOR FULL-LENGTH ARTICLES: This study systematically identified the effects of physical activity (PA) on depression, anxiety and weight-related outcomes among children and adolescents with overweight/obesity. EMBASE, The Cochrane Library, Web of Science, and PubMed were searched from January 1, 2000 to August 1, 2022 for peer-reviewed papers. Meta-analyses were conducted to ascertain the effect of physical activity on symptoms of anxiety, depression and weight-related outcomes in overweight/obese children and adolescents. Twenty-five studies representing 2188 participants, with median age 12.08 years old (8.3 to 18.44 years) were included. Depressive and anxiety symptoms, BMI, BMI z-scores, weight, waist circumference and height were evaluated. After incorporating the effects of PA interventions on children and adolescents with overweight/obesity, PA could improve depressive and anxiety symptoms, but not obesity indexes except waist circumference. While, PA combined with other interventions have a significant effect both on anxiety symptoms and BMI compared to pure PA intervention. In terms of intervention duration, we observed that durations falling within the range of 8 to 24 weeks exhibited the most positive effects on reducing depressive symptoms. FOR SHORT COMMUNICATIONS: We included 25 articles on the effects of physical activity on psychological states such as depression and anxiety, weight, BMI and other weight-related indicators in children and adolescents with overweight/obesity. We attempted to determine the most appropriate type of physical activity intervention for children and adolescents with overweight/obesity, as well as the most appropriate population characteristics and duration by combining the outcome data from each article. This has a great enlightening effect for health workers to carry out corresponding strategies in the future.

Pyruvate kinase deficiency and PKLR gene mutations: Insights from molecular dynamics simulation analysis.

Pyruvate kinase deficiency is a rare hereditary erythrocyte enzyme disease caused by mutations in the pyruvate kinase liver and red blood cell gene. The clinical presentations of pyruvate kinase deficiency are significantly heterogeneous, ranging from just mild anemia to hemolytic crisis or even death. The proband in our study was a 2-year-old girl for severe skin and scleral icterus with progressive aggravation. We collected the family's data for further analysis. Whole exome genome sequencing of the pedigree revealed a novel compound heterozygous mutation, c.1097del (p.P366Lfs*12) and c.1493G > A (p.R498H), in the pyruvate kinase liver and red blood cell gene. Furthermore, molecular dynamics simulations were employed to uncover differences between the wild type and mutant pyruvate kinase liver and red blood cell proteins, focusing on structural stability, protein flexibility, secondary structure, and overall conformation. The combined bioinformatic tools were also utilised to assess the effects of the missense mutation on protein function. Thereafter, wild type and mutant plasmids were constructed and transfected into 293T cells, and Western blot assay was conducted to validate the impact of the mutations on the expression of pyruvate kinase liver and red blood cell protein. The data presented in our study enriches the genotype database and provides evidence for genetic counseling and molecular diagnosis of pyruvate kinase deficiency.

The novel compound heterozygous variants identified in a Chinese family with glucose phosphate isomerase deficiency and pathogenicity analysis.

BACKGROUND AND AIMS: Glucose phosphate isomerase (GPI) deficiency is an extremely rare autosomal recessive disorder caused by mutations in the GPI gene. In this research, the proband displaying typical manifestations of haemolytic anaemia and his family members were recruited to analyse the pathogenicity of the detected variants. METHODS: Peripheral blood samples were collected from the family members and genomic DNA was extracted and targeted for capture and sequencing. The effect of the candidate pathogenic variants on splicing was further investigated using the minigene splicing system. The computer simulation was also used for further analysis of the detected data. RESULTS: The proband carried the compound heterozygous variants c.633 + 3 A > G and c.295G > T in the GPI gene, which have never been reported before. In the genealogy, co-segregation of the mutant genotype with the phenotype was established. The minigene study showed that intronic mutations resulted in abnormal pre-mRNA splicing. Specifically, the two aberrant transcripts: r.546_633del and r.633 + 1_633 + 2insGT were transcribed by the minigene plasmid expressing the c.633 + 3 A > G variant. The missense mutation c.295G > T in exon 3 resulted in altering glycine at codon 87 to cysteine which was predicted to be pathogenic in an in silico analysis. Deeper analyses revealed that the Gly87Cys missense mutation led to steric hindrance. Compared to the wild-type, the mutation G87C led to a significant increase in intermolecular forces. CONCLUSION: Overall, the novel compound heterozygous variants in the GPI gene contributed to the etiology of the disease. Genetic testing can assist in the diagnosis. The novel gene variants identified in the present study has further expanded the mutational spectrum of GPI deficiency, which can better guide family counselling.

The associations between hematological parameters and the incidence of prehypertension and hypertension in children and adolescents: a prospective cohort study.

Uncontrolled pediatric hypertension may increase the risk of hypertension in adulthood. Several studies have reported an association between hematological parameters and blood pressure (BP) levels. However, epidemiologic evidence of this association in children and adolescents remains scarce. This study aims to explore the associations between hematological parameters and the incidence of prehypertension and hypertension in children and adolescents. This longitudinal study was conducted with 1368 participants aged 6-8 years from baseline visit to follow-up visit. Compared with participants from the normal blood pressure (BP) group, participants from the elevated BP group had significantly higher baseline red blood cell (RBC) counts, hemoglobin (Hb) counts and hematocrit (Hct) levels (all P < 0.001). A multilevel linear mixed model was conducted to analyze the relationship between hematological parameters and BP levels. The results suggested that SBP, DBP and MAP increased significantly with a quartile increase of levels of hematological parameters (all P < 0.05). Furthermore, a multilevel mixed logistic regression model was used to analyze the risk of per interquartile range increase in hematological parameters on the incidence of prehypertension and hypertension. The risk of prehypertension and hypertension incidence increased by (1.34 (95%CIs: 1.20, 1.50)), (1.38 (95%CIs: 1.24,1.54)), (1.33 (95%CIs: 1.19,1.50)), (1.14 (95%CIs: 1.03,1.26)) fold with a one-quartile increase in levels of RBC, Hb, Hct and Fe, respectively (all P < 0.05). This longitudinal study showed that hematological parameters were positively associated with BP levels in healthy children and adolescents, which excluded the effect of antihypertensive drugs on BP levels that often appeared in adults.

Netrin-1 inducing antiapoptotic effect of acute myeloid leukemia cells in a concentration-dependent manner through the Unc-5 netrin receptor B-focal adhesion kinase axis.

Acute myeloid leukemia (AML) is a hematological malignancy that commonly occurs in children. The prognosis of pediatric AML is relatively poor, thus threatening the patient's survival. The aberrant expression of the axon guidance factor, netrin-1, is observed in various types of malignancies, and it participates in the proliferation and apoptosis of tumor cells. Herein, we aimed to explore the role of netrin-1 in AML cells. Netrin-1 is highly expressed in AML patients. Proliferation and anti-apoptosis were observed in AML cells treated with netrin-1. The interaction between netrin-1 and Unc-5 netrin receptor B (UNC5B) was detected through coimmunoprecipitation, and UNC5B ribonucleic acid interference restrained the influence of netrin-1 on the AML cells. The phosphorylation of focal adhesion kinase-protein kinase B (FAK-Akt) was upregulated in AML cells treated with netrin-1. Both FAK and Akt inhibitors abrogated the effects of netrin-1 on the proliferation and apoptosis of AML cells. In conclusion, netrin-1 could promote the growth and reduce the apoptosis of AML cells in a concentration-dependent manner, and that these effects were mediated by activating the FAK-Akt signaling pathway via the UNC5B.

Inhibition of Integrin αvß3-FAK-MAPK signaling constrains the invasion of T-ALL cells.

The role of adhesion receptor integrin αvß3 in T-ALL was unclear. Firstly, we performed quantitative real-time PCR to assess medullary expression of integrin ß3(ITGB3) in T-ALL patients and high ITGB3 expression was relevant with the central nervous system leukemia(CNSL) incidence. Decreasing of cell invasion was observed in Jurkat and Molt4 treated with integrin αvß3 specific antibody and inhibitor as well as cells with ITGB3 interference. Further, phosphorylation of FAK, cRAF, MEK and ERK decreased in cells with integrin αvß3 inhibition or interference. Invasion decreased in T-ALL cells treated with FAK and ERK inhibitors. In conclusion, inhibition of integrin αvß3 signals significantly limits the cell invasion of T-ALL cells.

De novo variations of ANK1 gene caused hereditary spherocytosis in two Chinese children by affecting pre-mRNA splicing.

BACKGROUND AND AIMS: Hereditary spherocytosis (HS) is one of the most common hereditary haemolytic disorders. Here, two unrelated families with the probands displaying typical manifestations of HS were enrolled. Our study aimed to characterize the effect of two novel variants in HS patients on gene splicing to help minimize the rate of misdiagnosis of HS and enhance clinicians' understanding of the disease. PARTICIPANTS AND METHODS: A retrospective review was conducted. Peripheral blood samples were collected from all the family members, and genomic DNA was extracted for genetic diagnostics. First, high-throughput sequencing technology was used for the preliminary screening of candidate causative variants. Thereafter, the variants were verified via Sanger sequencing. Furthermore, a pathogenicity analysis of the detected variants was performed including in silico prediction and in vitro experiments. We constructed matched wild-type and mutant-type minigene plasmid of ANK1 based on HEK293T cells to address the effects of variants on mRNA splicing. RESULTS: The c.1305 + 2 T > A (family1) and c.1305 + 2del (family2) variants were detected in the ANK1 gene. These two de novo mutations described by us which have not been reported prior to this study. Moreover, the validation results of splicing reporter systems revealed that the intronic mutations resulted in abnormal pre-mRNA splicing. Specifically, the minigene plasmid expressing the c.1305 + 2 T > A variant transcribed the two aberrant transcripts: r.1305_1306ins1305 + 1_1305 + 229 and r.1305_1306ins1305 + 1_1305 + 552. The minigene plasmid expressing c.1305 + 2del transcribed the two aberrant transcripts: r.1305_1306ins1305 + 1_1305 + 228 and r.1305_1306ins1305 + 1_1305 + 551. CONCLUSION: The two de novo variants identified in the ANK1 gene were the genetic etiology of the probands with HS in our study. Our findings further enrich the HS genotype database and provide a basis for genetic counselling and molecular diagnosis.

The effect of fine particulate matter exposure on allergic rhinitis of adolescents aged 10-13 years: A cross-sectional study from Chongqing, China.

Background: Allergic rhinitis (AR) has become a tremendous disease burden worldwide. Only a few studies have explored the effects of environmental exposure on the prevalence of AR in children in China. Methods: In the present study, we investigated the associations of environmental exposure (including fine particulate matter (PM2.5), air humidity, temperature, and passive smoking) with AR in adolescents aged 10-13 years in Chongqing. Data from 4,146 participants in urban and rural areas between March 2019 and May 2019 were collected. Results: The overall prevalence of AR was 17.50% in adolescents. After adjusting for other covariates, AR was positively correlated with the annual mean PM2.5 concentration, monthly mean PM2.5 concentration and air temperature, and negatively related to air humidity. Furthermore, the annual mean PM2.5 was positively associated with the risk of AR after adjusting for air temperature and humidity. Passive smoking (PS) was marginally associated with a high risk of AR. Conclusion: High PM2.5 exposure, high air temperature, and low air humidity were associated with a high risk of AR in adolescents. Our findings have potential implications for public health strategies and interventions aimed at reducing the burden of AR in adolescents.

Type O blood, the MCHC, and the reticulocyte count impact the early recurrence of primary warm-antibody autoimmune hemolytic anemia in children: A retrospective cohort analysis.

Objective: Primary warm-antibody autoimmune hemolytic anemia (w-AIHA) is prone to recurrence in children. In this study, we aimed to identify risk indicators for the early recurrence of primary w-AIHA and construct an effective recurrence risk assessment model. Methods: This was a retrospective cohort study. The clinical data of patients hospitalized with primary w-AIHA in the Department of Hematology and Oncology, Children's Hospital of Chongqing Medical University, between 1 January 2018 and 30 September 2021, were collected at the initial diagnosis. Univariate and multivariate logistic regression analyses were used to determine risk indicators for the early recurrence of primary w-AIHA in children, and ROC curve and Kaplan-Meier survival analyses were used for verification. Finally, a risk assessment model for early recurrence in children with primary w-AIHA was constructed using Cox regression and visualized using a nomogram. The model was also verified internally and externally. Results: This study included 62 children with primary w-AIHA. Of which, 18 experienced recurrence 1 year after the initial diagnosis. The univariate and multivariate logistic regression analyses showed that type O blood and the reticulocyte count (Ret) were risk indicators for the early recurrence of pediatric primary w-AIHA (P = 0.009, 0.047, respectively). The mean corpuscular hemoglobin concentration (MCHC) is a protective factor (P = 0.040). According to the ROC curve and Kaplan-Meier survival analyses, children with primary w-AIHA whose blood type was O or had an MCHC of <313.5 pg/fL or a Ret of ≥0.161×1012/L had a higher risk of early recurrence (HR = 2.640, 4.430 and 4.450, respectively, and P = 0.040, 0.015 and 0.018, respectively). The blood types (O), MCHCs, and Rets of 56 patients were incorporated into the Cox regression model, and the recurrence risk assessment model for children with primary w-AIHA was successfully constructed and visualized using a nomogram. The calibration curves and decision-curve analysis (DCA) suggested that the risk model has clinical applicability and effectiveness. Conclusion: Children with type O blood and an MCHC value of <313.5 pg/fL or a Ret value of ≥0.161×1012/L have a higher risk of early recurrence. The risk assessment model for the early recurrence of pediatric primary w-AIHA constructed in this study has good clinical applicability and effectiveness.

Netrin-1 induces the anti-apoptotic and pro-survival effects of B-ALL cells through the Unc5b-MAPK axis.

BACKGROUND: B-cell acute lymphoblastic leukemia (B-ALL) comprises over 85% of all acute lymphoblastic leukemia (ALL) cases and is the most common childhood malignancy. Although the 5 year overall survival of patients with B-ALL exceeds 90%, patients with relapsed or refractory B-ALL may suffer from poor prognosis and adverse events. The axon guidance factor netrin-1 has been reported to be involved in the tumorigenesis of many types of cancers. However, the impact of netrin-1 on B-ALL remains unknown. METHODS: The expression level of netrin-1 in peripheral blood samples of children with B-ALL and children without neoplasia was measured by enzyme-linked immunosorbent assay (ELISA) kits. Then, CCK-8 cell proliferation assays and flow cytometric analysis were performed to detect the viability and apoptosis of B-ALL cells (Reh and Sup B15) treated with exogenous recombinant netrin-1 at concentrations of 0, 25, 50, and 100 ng/ml. Furthermore, co-immunoprecipitation(co-IP) was performed to detect the receptor of netrin-1. UNC5B expression interference was induced in B-ALL cells with recombinant lentivirus, and then CCK-8 assays, flow cytometry assays and western blotting assays were performed to verify that netrin-1 might act on B-ALL cells via the receptor Unc5b. Finally, western blotting and kinase inhibitor treatment were applied to detect the downstream signaling pathway. RESULTS: Netrin-1 expression was increased in B-ALL, and netrin-1 expression was upregulated in patients with high- and intermediate-risk stratification group of patients. Then, we found that netrin-1 induced an anti-apoptotic effect in B-ALL cells, implying that netrin-1 plays an oncogenic role in B-ALL. co-IP results showed that netrin-1 interacted with the receptor Unc5b in B-ALL cells. Interference with UNC5B was performed in B-ALL cells and abolished the antiapoptotic effects of netrin-1. Further western blotting was applied to detect the phosphorylation levels of key molecules in common signaling transduction pathways in B-ALL cells treated with recombinant netrin-1, and the FAK-MAPK signaling pathway was found to be activated. The anti-apoptotic effect of netrin-1 and FAK-MAPK phosphorylation was abrogated by UNC5B interference. FAK inhibitor treatment and ERK inhibitor treatment were applied and verified that the FAK-MAPK pathway may be downstream of Unc5b. CONCLUSION: Taken together, our findings suggested that netrin-1 induced the anti-apoptotic effect of B-ALL cells through activation of the FAK-MAPK signaling pathway by binding to the receptor Unc5b. Video Abstract.

[Effect of Netrin-1 on VEGFA Expression in T-ALL Cells and Its Related Mechanism].

AbstractObjective: To investigate the effect of the axon guidance factor Netrin-1 on the expression of VEGFA in T cell acute lymphoblastic leukemia(T-ALL) and its related mechanism. METHODS: ELISA assays were applied to detect the levels of Netrin-1 and VEGFA in the bone marrow (BM) samples from children in the T-ALL and control group. The level of Netrin-1 and VEGFA were compared between control children and patients, and the liner correlation between Netrin-1 and VEGFA was analyzed. The T-ALL cells Jurkat and Molt-4 were culture in vitro, and the cells were treated with different concentration of Netrin-1 (0, 25, 50, 100 ng/ml) for 24 h, quantitative RT-PCR (qRT-PCR) and Western blot were used to detect the VEGFA expression in Jurkat, Molt-4 cells. The expression of Netrin-1 receptors in T-ALL cells was detected by qRT-PCR and the interaction between Netrin-1 and receptor in each cells was detected by co-IP. Furthermore, Western blot was used to detect the phosphorylation level of key prateins of AKT signal transduction pathway including Akt and mTOR in T-ALL cells treated with Netrin-1 (100 ng/ml). The expression of VEGFA and phosphorylation of AKT pathway transducers were detected by Western blot, after T-ALL cells treated with Netrin-1 (100 ng/ml) combined with inhibitors specific to Akt or mTOR. RESULTS: The expression level of Netrin-1 and VEGFA in T-ALL patients BM samples were both signi-ficantly higher than that of control group. And the expression level of Netrin-1 was positively correlated with that of VEGFA（r2=0974). With the increase of Netrin-1 concentration, the expression level of VEGFA also increased(P<0.05）. Netrin-1 interacted with its receptor, integrin-ß4 at the Netrin-1 concentration of 100 ng/ml. Further, the treatment of Netrin-1 could increase the phosphorylation of Akt and mTOR, which were the key transducers of AKT pathway. After treatment of T-ALL cells with Netrin-1 (100 ng/mL) and Akt inhibitor, the expression of VEGFA and phosphorylation of Akt or mTOR decreased. When the cells were treated with Netrin-1(100 ng/ml) and mTOR inbititor, the phosphorylation level of mTOR and the expression of VEGFA decreased, the phosphorylation level of Akt increased. CONCLUSION: The expression of Netrin-1 and VEGFA in bone marrow of childred with T-ALL were abnormal, and there was a linear relationship between them. Netrin-1 can interact with its receptor, integrin-ß4 and activate AKT transduction pathway to elevate the expression of VEGFA in T-ALL cells.

Combining multiaspect factors to predict the risk of childhood hypertension incidence.

Childhood hypertension has become a global public health issue due to its increasing prevalence and association with cerebral-cardiovascular disease in adults. In this study, we developed a predictive model for childhood hypertension based on environmental and genetic factors to identify at-risk individuals. Eighty children diagnosed with childhood hypertension and 84 children in the control group matched by sex and age from an established cohort were included in a nested case-control study. We constructed a multiple logistic regression model to analyze the factors associated with hypertension and applied the 10-fold cross-validation method to verify the prediction stability of the model. Childhood hypertension was found positively correlated with triglyceride level ≥150 mg/dL; low-density lipoprotein cholesterol level ≥110 mg/dL; body mass index Z score; waist-to-height ratio Z score; and red blood cell count (all P < .01) and negatively correlated with the relative expression level of retinol acyltransferase; relative expression level of vitamin D receptor; and dietary intake of fiber, vitamin C and copper (all P < .05) in this study. BMI Z score, triglyceride ≥150 mg/dL, RBC count, VDR/ß-actin and LRAT/ß-actin ratios were used to construct the predictive model. The area under the receiver operating characteristic curve was 94.45% (95% CI = 89.35%∼98.65%, P < .001). The accuracy, sensitivity, specificity, positive predictive value, and negative predictive value were all above 80% in both the training and validation sets. In conclusion, this model can predict the risk of childhood hypertension and could provide a theoretical basis for early prevention and intervention to improve the cardiovascular health of children.

The Impact of PM2.5 on the Growth Curves of Children's Obesity Indexes: A Prospective Cohort Study.

Aims: To explore the effect of long-term exposure to particulate matter with an aerodynamic diameter of 2.5 µm or less (PM2.5) on childhood obesity based on a cohort study in Chongqing. Methods: A total of 4,284 children aged 6-8 years at baseline were enrolled from the Chongqing Children Health Cohort in 2014-2015 and were followed up in 2019. A stratified cluster sampling was applied to select the participants. A Mixed-effects linear regression model was used to examine the effect of long-term exposure to PM2.5 on the growth curve of obesity indicators [including body mass index (BMI), BMI Z-score (BMIz), and waist-to-height ratio (WHtR)]. A mixed-effects logistic regression model was used to study the dose relationship between PM2.5 exposure and the risk of obesity indicators. Results: A higher level of accumulating exposure to PM2.5 was associated with an increased childhood obesity index, and the effect was the most significant for WHtR than BMI and BMIz. This effect was more pronounced in boys than in girls except for WHtR, and it was the most significant under the PM2.5 exposure period from pregnancy to 6 years old. Compared the annual average PM2.5 exposure level of <60 µg/m3, the WHtR and BMI were increased by 0.019 [(95% CIs): 0.014, 0.024] and 0.326 [(95% CIs): 0.037, 0.616] Kg/m2 for participants living with the PM2.5 exposure level of 70-75 µg/m3, respectively. For every 5 µg/m3 increase in PM2.5 levels (from pregnancy to 6 years old), the risk of central obesity was increased by 1.26 {odds ratio [OR] (95% CIs): 1.26 (1.16, 1.37), p < 0.001} times. Conclusions: This study confirmed a dose-response relationship between PM2.5 exposure and childhood obesity, especially central obesity, suggesting that controlling ambient air pollution can prevent the occurrence of obesity in children and adolescents.

The impact of PM2.5 on children's blood pressure growth curves: A prospective cohort study.

OBJECTIVE: The aim of this study was to explore the association between exposure to particulate matter with an aerodynamic diameter of 2.5 mm or less (PM2.5) and blood pressure (BP) levels in children and adolescents and to illustrate the impact of PM2.5 levels on BP growth curves in a cohort study. METHODS: A longitudinal study was designed and included 4303 children (7617 BP measurements) living in the selected areas, and evaluations were conducted in 2014-2015 (visit 1) and followed up in 2019 (visit 2). Two-stage stratified cluster sampling was used to include urban-rural areas. A mixed linear regression model and mixed logistic regression model were used to analyze the effect of PM2.5 exposure on BP and the incidence of prehypertension and hypertension in children. RESULTS: After adjusting for covariates, systolic blood pressure (SBP) (2.21 (95% CIs: 0. 81, 3.62), mmHg), diastolic blood pressure (DBP) (1.92 (95% CIs: 0.74, 3.11), mmHg), mean arterial pressure (MAP) (2.03 (95% CIs: 0.89, 3.17), mmHg) and heart rate (HR) (2.24 (95% CIs: 0.11, 4.37), beats/min) increased significantly in the fourth quartile of PM2.5 exposure levels compared with the first quartile (all P < 0.01). In addition, long-term exposure to PM2.5 was significantly positively correlated with SBP, DBP and MAP, and the effect was more notable in urban areas than that in rural areas. Moreover, the risk of prehypertension and hypertension incidence increased by 1.17 (95% CIs: 1.03, 1.33) fold with a one-quartile increase in PM2.5 exposure. The long-term effects of annual mean PM2.5 exposure on SBP, DBP and MAP were significant from pregnancy to 7, 3 and 4 years of age, respectively. CONCLUSION: Long-term exposure to PM2.5 was positively associated with growth curves of hemodynamics indexed from pregnancy to childhood and adolescence, and the effect was more significant in urban areas than in rural areas.

Prognostic significance of measurable residual disease based on multiparameter flow cytometry in childhood acute myeloid leukemia. / åŸºäºŽæµå¼ç»†èƒžæœ¯çš„å¯æµ‹é‡æ®‹ç•™ç— å¯¹å„¿ç«¥æ€¥æ€§é«“ç³»ç™½è¡€ç— é¢„åŽæ„ä¹‰çš„å‰çž»æ€§ç ”ç©¶.

OBJECTIVES: To study the prognostic value of measurable residual disease (MRD) for childhood acute myeloid leukemia (AML) by analyzing MRD-guided risk stratification therapy. METHODS: A total of 93 children with AML were prospectively enrolled in this study. Chemotherapy with the 2015-AML-03 regimen was completed according to the risk stratification determined by genetic abnormality at initial diagnosis and MRD and bone marrow cytology after induction therapy I. Multiparameter flow cytometry was used to dynamically monitor MRD and analyze the prognostic effect of MRD on 3-year cumulative incidence of recurrence (CIR) rate, event-free survival (EFS) rate, and overall survival (OS) rate. RESULTS: The 93 children with AML had a 3-year CIR rate of 48%±6%, a median time to recurrence of 11 months (range 2-32 months), a 3-year OS rate of 65%±6%, and a 3-year EFS rate of 50%±5%. After induction therapy I and intensive therapy I, the MRD-positive children had a significantly higher 3-year CIR rate and significantly lower 3-year EFS and OS rates than the MRD-negative children (P<0.05). There were no significant differences in 3-year CIR, EFS, and OS rates between the MRD-positive children with a low risk at initial diagnosis and the MRD-negative children after adjustment of chemotherapy intensity (P>0.05). The multivariate analysis showed that positive MRD after intensive treatment I was a risk factor for 3-year OS rate in children with AML (P<0.05). CONCLUSIONS: MRD has predictive value for the prognosis of children with AML. Based on the MRD-guided risk stratification therapy, reasonable application of chemotherapy may improve the overall prognosis of children with AML.

[Bioinformatics Analysis and Verification of Acute B-Lymphocytic Leukemia in Children with Isolated Bone Marrow Relapse].

OBJECTIVE: To analyze the hub genes affecting the solely bone marrow relapse of childish acute B-cell lymphoblastic leukemia (B-ALL). METHODS: The high-throughput RNA sequencing data were downloaded from TCGA database, the differentially expressed genes were screened by DESeq2 package of R, and the differentially expressed genes were grouped by GO function enrichment analysis and KEGG pathway enrichment analysis. Further, the data of STRING database and Cytoscape software were used to construct protein interaction network, screen hub genes and highly interaction protein sub network, perform GO and KEGG analysis of the hub genes and protein sub network respectively. JASPAR database was used to screen the upstream transcription factor of the hub gene promoter. Survival analysis based on the expression of hub genes was performed with clinical information attached to TCGA database. The bone marrow samples and clinical data of the patients were collected, the analysis results of hub genes were verified through clinical samples. RESULTS: 847 differentially expressed genes were collected, including 813 up-regulated genes, 34 down-regulated genes, 11 hub genes were screened out. The results of survival analysis showed that RPS5、RPS15、RPL23、RPL35、RPS8、RPS27A、RPS3、RPL9、RPS21、RPS7 and RPL38 showed significant effect on the survival of the children, and ZNF460 might be involved in their regulation. The high expressions of RPS3, RPS15, RPS8, RPS27A, and RPS21 had been verified in clinical samples of solely bone marrow relapsed patients. CONCLUSION: RPS3, RPS15, RPS8, RPS27A, RPS21 can be used as biomarkers to indicate the malignant event of solely bone marrow relapse, which may be regulated by ZNF460.

[Clinical features and prognosis of pediatric acute megakaryocytic leukemia].

OBJECTIVE: To study the clinical features and prognosis of children with acute megakaryocytic leukemia (AMKL) and the clinical effect of acute myeloid leukemia 03 (AML03) regimen for the treatment of pediatric AMKL. METHODS: The clinical data were collected from 47 children with AMKL who were diagnosed from May 2011 to December 2019. The treatment outcomes and prognostic factors were analyzed. The Kaplan-Meier method and the log-rank test were used for survival analysis. RESULTS: Among the 47 children with AMKL, 22 with non-Down syndrome-AMKL were treated by the AML03 regimen, with a median follow-up time of 11.4 months. For the 22 non-Down syndrome-AMKL patients, the remission rate of bone marrow cytology was 85% and the negative rate of minimal residual disease (MRD) was 79% after induction Ⅱ, with a 2-year overall survival (OS) rate of (50±13)% and a 2-year event-free survival (EFS) rate of (40±12)%. The group with positive immunophenotypic marker CD56 had significantly lower 2-year EFS and OS rates than the group with negative CD56 (P < 0.05). The group without remission of bone marrow cytology after induction Ⅱ had significantly lower 2-year EFS and OS rates than the group with remission (P < 0.05). The group with positive MRD after induction Ⅱ had a significantly lower 2-year EFS rate than the group with negative MRD (P < 0.05). There was no significant difference in 2-year OS and EFS rates between the patients with transplantation and those without transplantation (P > 0.05). CONCLUSIONS: Children with AMKL tend to have a low remission rate and a poor prognosis. Positive immunophenotypic marker CD56, bone marrow cytology during early treatment response, and MRD results are important factors influencing the prognosis. Allogeneic hematopoietic stem cell transplantation has no significant effect on the prognosis of AMKL.

[Expression of Netrin-1 in Patients with Acute Lymphoblastic Leukemia and Its Clinical Significance].

OBJECTIVE: To investigate the correlation of the Netrin-1 expression level with the clinical characteristics in children with acute lymphoblastic leukemia (ALL) and to explore its possible regulatory mechanism. METHODS: ELISA was used to detect the expression level of Netrin-1 in peripheral blood serum from 48 child ALL patients (newly diagnosed, recurrent), and its relevance with clinical indicators was statistically analyzed. The blood serum samples from 27 children with non malignant hematological diseases were choosen as controls. Leukemia cell lines of Jurkat,Molt-4,SUP-B15 and Raji were cultivated in vitro, after treated with different concentrations of recombinant human Netrin-1 protein， the invasive ability of the cells was detected by Transwell method； the effect of Netrin-1 to the proli feration of cells was detected by CCK-8 method； The expression and phosphorylation level of key molecules, such as FAK,Erk1/2,PI3K and Akt signaling pathway were detected by Western blot. RESULTS: The expression of Netrin-1 in child patients was significantly higher than that of the control group (P<0.05). With the increasing of Netrin-1 level, the level of Plt (r=0.483, P<0.05) increased, while the level of WBC (r=-0.290, P<0.05) decreased, and there were no significant correlation with age, Hb level and the proportion of immature cells in bone marrow. When the concentration of Netrin-1 was 25-50 ng/ml, the level of Netrin-1 positively correlated with WBC (r=0.886, P<0.05) ; the level of Netrin-1 significantly decreased when the patient's WBC was ＞50×109/L and Plt ＞20×109/L(P=0.042，P=0.001); The expression level of Netrin-1 was significantly different in the risk group(P=0.017), and level of Netrin-1 in high-risk group was significantly higher than that in low risk group and middle risk group, but there was no significant difference of Netrin-1 expression in sex, hepatosplenomegaly, MRD, recurrence and chromosome abnormality. Netrin-1 could promote the invasiveness of the four kinds of cells (P<0.05). With the increase of Netrin-1 concentration, the number of cells increased at first and then decreased, and the number of cells in the invading chamber was the highest when the concentration of Netrin-1 was 100 ng/ml; the survival rate of the four kinds of cells significantly increased when the concentration of Netrin-1 was 25 ng/ml(P<0.05), and SUP-B15 cells showed the highest cell survival rate at a concentration of 100 ng/ml; The survival rate of the four kinds of cells showed a tendency : survival of cells increased at low concentration of Netrin-1 and survival of cells decreased at high concentration of Netrin-1. The results of Western blot showed that Netrin-1 activated the phosphorylation level of key molecules such as FAK,Erk1/2,PI3K,Akt signaling pathway (P<0.05). CONCLUSION: There is abnormal expression of Netrin-1 in serum of children with ALL. Netrin-1 may affect the occurrence and development of ALL by increasing the proliferation and invasiveness of leukemia cells, and may become a risk factor of ALL or a potential target in biotherapy.

Liraglutide protects pancreatic ß cells from endoplasmic reticulum stress by upregulating MANF to promote autophagy turnover.

AIMS: This study was conducted to determine the relationship between mesencephalic astrocyte-derived neurotrophic factor (MANF), autophagy and endoplasmic reticulum (ER) stress, and whether liraglutide (LRG) can protect ß cells, promote autophagy and alleviate ER stress by regulating MANF expression. MAIN METHODS: Human serum samples were collected from healthy controls (NC), simple hyperlipidemia (HLD), and newly diagnosed type 2 diabetes (T2D). The MANF levels were detected using ELISA. In vitro, after the mouse islet MIN6 cells were treated with glucose (GLU), palmitate (PA), thapsigargin (TG), LRG, and chloroquine (CQ), cell proliferation was detected using cell counting kit-8 (CCK-8), apoptosis-related protein cleaved caspase 3 (C-cas-3), ER stress, and autophagy-related proteins were detected by Western blotting, MANF, insulin, and C-cas-3 proteins were detected via immunofluorescence. Subcellular structures and autophagosomes were examined using electron microscopy. KEY FINDINGS: Compared with the NC group, the MANF levels in the HLD and T2D groups increased significantly. After ER stress induced by GLU, PA, and TG, cell viability decreased, while MANF, c-cas3, ERS, and autophagy-related proteins increased, which was related to the concentration of GLU, PA, and TG. Compared with the BSA group, the number of mitochondria and autophagosomes in the PA group increased and the mitochondria were damaged. In the PA and TG plus CQ groups, the effect was further exaggerated. But after co-treatment with LRG, the effects of GLU, PA, and TG were attenuated. SIGNIFICANCE: LRG protects islet ß cells from ER stress by upregulating MANF to promote autophagy turnover.

Clinical efficacy and safety of imatinib treatment in children and adolescents with chronic myeloid leukemia: A single-center experience in China.

Chronic myeloid leukemia (CML) is relatively rare in childhood and few studies have reported the clinical use of imatinib (IM) in pediatric CML. In this study, we evaluated the efficacy and tolerability of IM in children and adolescents with CML.We investigated 21 patients under 18 years of age with newly diagnosed CML and treated with IM in Children's Hospital of Chongqing Medical University between May 2014 and February 2018. The disease was staged according to the European LeukemiaNet criteria and the IM dose was determined based on the disease stage. Cumulative responses and survival probabilities were estimated according to the Kaplan-Meier method.The estimated complete hematologic response rate of chronic phase-chronic myeloid leukemia (CML-CP) was 89.5% at 3 months. The complete cytogenetic response rates increased with time, reaching 47.4%, 73.7%, and 80.3% at 6, 12, and 24 months, respectively. The cumulative major molecular response rates were 42.1% and 76.3% at 12 and 24 months, respectively. With a median follow-up time of 33.8 months (range, 3.2-61.7 months), the estimated 2-year overall survival (OS) rate for CML was 95.2% (95% confidence interval [CI], 70.7%-99.3%). None of the CML-CP patients progressed to the accelerated phase or had a blast crisis. The 2-year OS and progression-free survival rates for the CML-CP cohort were both 100%, while the estimated 2-year event-free survival rate was 68% (95% CI, 42.1%-84.2%). None of the patients in this group had treatment-related deaths or IM discontinuation due to drug toxicities, and only 1 patient had a grade III-IV nonhematologic adverse event. Overall, anemia was the most common adverse effect and 42.9% of patients had a decrease in bone mineral density.IM was effective and the adverse effects were well-tolerated throughout the follow-up period in Chinese CML patients under 18 years of age.