This study investigated the intracellular localization of asparagine synthetase (ASNS) in the relation with chemoresistance in leukemia. pIRES-GFP-ASNS-Flag/Neo expression vector was transiently tansfected into SK-N-MC cells and 297T cells respectively. Immunofluorescence and Western blot analysis were performed for cellular localization of ASNS respectively. U937 cells were treated with L-asparaginase for 48 h and examined for endogenous ASNS expression on plasma membrane by immunofluorescence staining. Immunofluorescence staining showed that the transiently expressed ASNS was partly localized on transfected-SK-N-MC cell surface. Moreover, Western blotting exhibited that ASNS expressed both in cytosol and on plasma membrane of transfected-293T cells. Immunofluorescence staining with anti-ASNS-specific monoclonal antibody revealed that endogenous ASNS was localized on the plasma membrane of U937 cells, except for its distribution in the cytosol. In addition, ASNS exhibited a higher expression on plasma membrane after treatment with L-asparaginase as compared with the untreated cells. It was concluded that the subcellular translocation of ASNS may play an important role in L-asparaginase resistance in leukemia cells.

Objective To investigate the benefits of replanning after induction chemotherapy(IC) by analyzing the dosimetric impact of IC on intensity-modulated radiotherapy(IMRT)for locally advanced nasopharyngeal carcinoma(NPC)and the dosimetric characteristics of replanning after IC, and to provide data for the rational design of clinical radiotherapy plans. Methods 16 NPC patients underwent contrast-enhanced CT scan once before and after IC.Target volumes were delineated and the chemotherapy plans were created,defined as Plan-1 and Plan-2,respectively. Then the target structure after IC was copied to Plan-1, generating the third plan, defined as Plan-1-2. The paired t-test was used to compare the dosimetric parameters between Plan-1 and Plan-1-2 and between Plan-2 and Plan-1-2. Results Plan-1 vs. Plan-1-2:Plan-1-2 showed significantly reduced D meanof target volume compared with Plan-1(P<0.05). Plan-1-2 significantly increased D meanand D maxof the spinal cord(P<0.05),although significantly reduced D mean of the brain stem and D maxof the temporal lobes compared with Plan-1. Plan-1-2 also had significantly reduced conformity index(CI)and significantly increased homogeneity index(HI)for the target volume compared with Plan-1(P<0.05). Plan-2 vs. Plan-1-2:Compared with Plan-1-2, Plan-2 significantly increased D meanand D minof gross tumor volume(GTV)and primary GTV(P<0.05)and significantly reduced D meanof the temporal lobes and D maxand D meanof the spinal cord(P<0.05), with D max decreased to 430.48 cGy;Plan-2 had significantly increased CI and significantly reduced HI for the target volume compared with Plan-1-2(all P<0.05). Conclusions IMRT plan-1 after IC has worse dosimetric distribution,while replanning after IC has more dosimetric benefits.

This study investigated the intracellular localization of asparagine synthetase (ASNS) in the relation with chemoresistance in leukemia.pIRES-GFP-ASNS-Flag/Neo expression vector was transiently tansfected into SK-N-MC cells and 297T cells respectively.Immunofluorescence and Western blot analysis were performed for cellular localization of ASNS respectively.U937 cells were treated with L-asparaginase for 48 h and examined for endogenous ASNS expression on plasma membrane by immunofluorescence staining.Immunofluorescence staining showed that the transiently expressed ASNS was partly localized on transfected-SK-N-MC cell surface.Moreover,Western blotting exhibited that ASNS expressed both in cytosol and on plasma membrane of transfected-293T cells.Immunofluo-rescence staining with anti-ASNS-specific monoclonal antibody revealed that endogenous ASNS was localized on the plasma membrane of U937 cells,except for its distribution in the cytosol.In addition,ASNS exhibited a higher expression on plasma membrane after treatment with L-asparaginase as compared with the untreated cells.It was concluded that the subcellular translocation of ASNS may play an important role in L-asparaginase resistance in leukemia cells.

OBJECTIVEMucopolysaccharidosis(MPS) is a congenital hereditary disease. Only a few patients with this disease can be controlled by enzyme replacement therapy. Most of them are short of effective interference. To exploit the effect of treatment with allogenic hematopoietic stem cell transplantation, two children were treated with the transplantation.

METHODSThe two patients included a 23 month MPS-IH and an 18 month old MPS-VI at the time of transplantation. Busulfan of 20 mg/kg plus 200 mg of Cyclophosphamide were used as the conditioning regimen. Peripheral stem cells were collected from a 9/10 high resolution matched unrelated donor and a matched sibling carrier donor, respectively. The heart and lung were affected in the patient with MPS-IH. Medium obstructed pulmonary impairment was found by pulmonary function test at the time of transplantation. Medium mitral valve countercurrent and patent ductus arteriosis(PDA) were found by Doppla examination.

RESULTSThe number of hematopoietic stem cells was comparative between the two donors with total nucleated cells and CD34+ cells of 11 x 10(8)/kg and 17 x 10(8)/kg, and 7.6 x 10(6)/kg and 7.2x 10(6)/kg respectively. Neutrophil engrafted at day 11. The process of transplantation in the MPS-VI patient went smoothly with grade II graft versus host disease(GVHD) briefly and only 1 U RBC and 2 U platelet were transfused. For the MPS-IH patient, the process of transplantation was tough with platelet reaching to 20 x 10(9)/L till day 40 and 5 U RBC and 7 U platelet were transfused during transplantation. Grade III GVHD was resolved by steroid, mycophenolate mofetil (MMF) and CD25 antibody. Pneumonia recurred 3 times with 2 times rescued by trachea intubation and mechanical ventilation because of accompanying acute heart failure. At day 14 the lymphocytes in both patients were 100% from donors as evidenced by short tandem repeat-PCR(STR-PCR). MPS associated enzyme activity was increased to 70 nmol/h.mg and 66 nmol/h.mg at 3 month and still remained 50.9 nmol/h.mg and 44.5 nmol/h.mg at 2 years post transplantation. Till now the 2 patients have been followed up for 25 months and 28 months with good general condition. The cardiac and pulmonary functions have improved obviously in the MPS-IH patient. The cornea became clear in this patient.

CONCLUSIONAllogeneic hematopoietic stem cell transplantation is an effective measure to treat patient with MPS-IH and MPS-VI. Transplantation at earlier stage of age can decrease transplant related complications. It requires longer time follow up for observing the clinical effects for these patients.

Female ; Follow-Up Studies ; Graft vs Host Disease ; drug therapy ; etiology ; Hematopoietic Stem Cell Transplantation ; adverse effects ; methods ; Humans ; Infant ; Intraoperative Complications ; drug therapy ; etiology ; Male ; Mucopolysaccharidoses ; enzymology ; pathology ; physiopathology ; surgery ; Recovery of Function ; Transplantation, Homologous

Objective:To explore the feasibility and validity of constructing an intensity-modulated radiotherapy gamma pass rate prediction model after combining the SHAP values with the extreme gradient boosting tree (XGBoost) algorithm feature selection technique, and to deliver corresponding model interpretation.Methods:The dose validation results of 196 patients with pelvic tumors receiving fixed-field intensity-modulated radiotherapy using modality-based measurements with a gamma pass rate criterion of 3%/2 mm and 10% dose threshold in Hunan Provincial Tumor Hospital from November 2020 to November 2021 were retrospectively analyzed. Prediction models were constructed by extracting radiomic features based on dose files and using SHAP values combined with the XGBoost algorithm for feature filtering. Four machine learning classification models were constructed when the number of features was 50, 80, 110 and 140, respectively. The area under the receiver operating characteristic curve (AUC), recall rate and F1 score were calculated to assess the classification performance of the prediction models.Results:The AUC of prediction model constructed with 110 features selected based on the SHAP-valued features was 0.81, the recall rate was 0.93 and the F1 score was 0.82, which were all better than the other 3 models.Conclusion:For intensity-modulated radiotherapy of pelvic tumor, SHAP values can be used in combination with the XGBoost algorithm to select the optimal subset of radiomic features to construct predictive models of gamma pass rates, and deliver an interpretation of the model output by SHAP values, which may provide value in understanding the prediction by machine learning-dependent models.