LATS1 Promotes B-ALL Tumorigenesis by Regulating YAP1 Phosphorylation and Subcellular Localization

Objective YAP1 plays a dual role as an oncogene and tumor suppressor gene in several tumors;differentiating between these roles may depend on the YAP1 phosphorylation pattern.The specific function of YAP1 in B cell acute lymphoblastic leukemia(B-ALL),however,is currently unclear.Thus,in the present study,the role of YAP1 in B-ALL was investigated using relevant cell lines and patient datasets.Methods The effects of shRNA-mediated knockdown on YAP1 and LATS1 levels in the NALM6 and MOLT-4 cell lines were examined using Western blotting,quantitative real-time polymerase chain reaction,flow cytometry,immunostaining,and nude mouse subcutaneous tumorigenesis experiments.Gene expression levels of Hippo pathway-related molecules before and after verteporfin(VP)treatment were compared using RNA-Seq to identify significant Hippo pathway-related genes in NALM6 cells.Results Patients with ALL showing high YAP1 expression and low YAP1-Ser127 phosphorylation levels had worse prognoses than those with low YAP1 protein expression and high YAP1-Ser127 phosphorylation levels.YAP1-Ser127 phosphorylation levels were lower in NALM6 cells than in MOLT-4 and control cells;YAP1 was distributed in the nuclei in NALM6 cells.Knockdown of YAP1 inhibited MOLT-4 and NALM6 cell proliferation and arrested the NALM6 cell cycle in the G0/G1 phase.Before and after VP treatment,the expression of the upstream gene LATS1 was upregulated;its overexpression promoted YAP1-Ser127 phosphorylation.Further,YAP1 was distributed in the plasma.Conclusion LATS1 may downregulate YAP1-Ser127 phosphorylation and maintain B-ALL cell function;thus,VP,which targets this axis,may serve as a new therapeutic method for improving the outcomes for B-ALL patients.

Process optimization of a closed-chamber distillation system for the recovery of FLiNaK molten salt

Low-pressure distillation has been proposed as a suitable technique for the recovery of carrier salt from molten salt reactor spent fuel.A closed-chamber distillation system,in which the pump is stopped and pressureinduced salt distillation is performed,was arranged for fluoride salt treatment.A stair-step optimization process was demonstrated to improve the recovery efficiency by up to 99%.The pressure change curve was feasible for estimating the distillation process,and a method for displaying the pressure value online in order to determine the endpoint was also developed.The decontamination factor of Nd in the condensate salt was deduced to be greater than 100 with 1 wt%NdF3–FLiNaK distillation.The optimal conditions developed in this study showed a high recovery ratio for the fluoride carrier salt and a high separation efficiency for rare earth products.

Genistein-induced Anticancer Effects on Acute Leukemia Cells Involve the Regulation of Wnt Signaling Pathway Through H4K20mel Rather Than DNA Demethylation

Objective:To investigate the effects and mechanisms of genistein on the gene expression in the Wnt pathway in acute leukemia(AL)cells.Methods:The expression of Wnt pathway genes and cell cycle-related genes were analyzed in two AL cell lines.Pyrophosphate sequencing was performed to determine the methylation degree.Then,the enrichment of H4K20mel and H3K9ac was determined using ChIP-qPCR.Flow cytometry was used to analyze the cell cycle.Results:The IC_(50) of genistein in the two AL cell lines was lower than that for the bone marrow mesenchymal stem cell line.Genistein upregulated H4K20mel,KMT5A and Wnt suppressor genes,including Wnt5a,and downregulated the downstream target genes of Wnt,such as c-myc and β-catenin.The methylation degree and H3K9ac enrichment in the Wnt5a promoter region remained unchanged.However,the enrichment of H4K20mel in the Wnt5a promoter and coding regions increased.In addition,genistein upregulated Phospho-cdc2,Mytl,Cyclin A,Cyclin E2,p21 and Phospho-histone H3,but downregulated Phospho-weel.Cell cycle arrest was induced in the G2/M phase.Conclusion:Genistein inhibits the activation of the Wnt pathway by promoting the expression of Wnt5a through the activation of KMT5A and enrichment of H4K20mel in the Wnt5a gene promoter and coding regions,rather than demethylation.Genistein also blocks the cell cycle in the G2/M phase.Therefore,genistein is a potential anti-leukemia drug.

Unsteady state precipitation of M_(23)C_(6)carbides during thermal cycling in reduced activation steel manufactured by laser melting deposition

The temperature field distribution and thermal history of Fe-9Cr2WVTa reduced activation steel prepared by laser melting deposition(LMD)have been calculated with Gaussian and Ring laser beams,and the nucleation and growth behaviors of M_(23)C_(6)precipitates in the 1st,7th and 19th layers have been calculated using the modified classical nucleation theory and Svoboda Fischer Fratzl Kozeschnik model.The energy distribution shows W-shape with Ring laser beam while it shows V-shape with Gaussian laser beam,which results in the more uniform M_(23)C_(6)size in the same layer with Ring laser beam.Precipitates in the bottom(i.e.,the 1st layer)have the minimum size and the size increases with the layer number with Gaussian and Ring laser beams.The temperature history,the instantaneous nucleation rate and the size evolution of M_(23)C_(6)have been systematically discussed.The results indicate that the nucleation,growth and re-dissolution of precipitates in reduced activation steel depend on the amount of energy absorbed in the thermal cycle during LMD.The continuous accumulation of energy during the thermal cycle leads to larger M_(23)C_(6)at the top area.The unsteady state precipitation dynamics of M_(23)C_(6)carbides during thermal cycling are consistent with the simulation results.