Objective:To explore the effect of intervention of E-cadherin (E-cad) and B-lymphoma Moloney murine leukemia virus insertion region-1 (Bmi-1) mediated by transcription activator-like effector nuclease (TALEN) on the biological behaviors ofnasopharyngeal carcinoma cells.Methods:Multi-locus gene targeting vectors pUC-DS1-CMV-E-cad-2A-Neo-DS2 and pUC-DS1-Bmi-1 shRNA-Zeo-DS2 were constructed,and the E-cad and Bmi-1 targeting vectors were transferred with TALEN plasmids to CNE-2 cells individually or simultaneously.The integration of target genes were detected by PCR,the expressions of E-cad and Bmi-1 were detected by Western blot.The changes of cell proliferation were detected by cell counting kit-8 (CCK-8) assay.T-he cell cycle and apoptosis were detected by flow cytometry.The cell migration and invasion were detected by Transwell assay.Results:The E-cad and Bmi-1 shRNA expression elements were successfully integrated into the genome of CNE-2 cells,the protein expression level of E-cad was up-regulated,and the protein expression level of Bmi-1 was down-regulated.The intervention of E-cad and Bmi-1 didn't affect the proliferation,cell cycle and apoptosis of CNE-2 cells,but it significantly inhibited the migration and invasion ability of CNE-2 cells.Furthermore,the intervention of E-cad and Bmi-1 together significantly inhibited the migration ability of nasopharyngeal carcinoma cells compared with the intervention of E-cad or Bmi-1 alone (all P＜0.01).Conclusion:The joint intervention of E-cad and Bmi-1 mediated by TALEN can effectively inhibit the migration and invasion of nasopharyngeal carcinoma cells in vitro,which may lay the preliminary experimental basis for gene therapy of human cancer.

Objective To observe dosage of insulin and analysis of glucose metabolism characteristics in the patients during the process of diabetic nephropathy (DN).Methods A total of 84 cases of diabetic nephropathy admitted into the Department of Nephrology of the hospital for treatment from January 2013 to December 2015 were chosen for the study.In accordance with the chronic kidney disease ( CKD) stages standards, as specified in the National Kidney Foundation′s Kidney Disease Treatment Guidelines, the DN process in the same patient was divided into 5 different stages:stage 1 ( CKD phase Ⅰand Ⅱ) , stage 2 ( CKD phase Ⅲ and Ⅳ) , stage 3 ( CKD phaseⅤand prior to dialysis therapy) , stage 4 ( CKD phase Ⅴand dialysis therapy for 1 to 3 months) , stage 5 ( CKD phase Ⅴand dialysis therapy for over 6 months) .Data pertaining to body mass, body mass index( BMI) , dosage of insulin and hemo-globin A1c(HbA1c), glomerular filtration rate (GFR), hemoglobin(Hb), fasting plasma glucose(FBG), postprandial blood glucose ( PBG) after 2 hours and albumin( ALB) were collected during the 5 stages, then were compared and analyzed statistically.Results During the first 3 stages, there were significant differences in the levels of GFR, Hb and ALB, as well as in dosage of insulin in the 84 patients(P<0.01), and the levels of GFR, Hb and ALB were all significantly higher than those of the second and third stages(P<0.01).The dosage of insulin during the first stage was (0.62 ±0.20) U/kg, which was significantly higher than those of the second and third stages〔(0.44 ±0.17) U/kg, (0.42 ±0.20) U/kg〕.However, no statistical significance could be seen in the dosage of in-sulin in the patients during the second and third stages(P>0.05).There were significant differences in the levels of GFR, Hb, ALB, and the dosage of insulin during the third, fourth and fifth stages for the 84 patients(P<0.01 or P<0.05).And the dosage of insulin during the third stage was significantly higher than those of the fourth and fifth stages (P<0.01).However, no statistical significance could be noted in the dosage of insulin during the fourth and fifth stages (P>0.05).The rates of hypoglycemia during the fourth and fifth stages were respectively 10 cases and 11 cases, which were all significantly higher than that of the third stage (2 cases) (P<0.05).Conclusion When the disease status of the DN patients developed from CKD Ⅰ~Ⅱphase to CKD Ⅲ~Ⅳ, the dosage of in-sulin should be reduced, and when it developed from CKD Ⅲ~Ⅳto CKD Ⅴ before the implementation of dialysis, the dosage could still remain at relatively stable level.However, when dialysis therapy was implemented for 1 to 3 months, the dosage of insulin should further be reduced.From then onwards, the dosage of insulin could remain basically unchanged.At the same time, for those patients with DN, measures should be taken for the prevention of hypoglycemia during dialysis therapy.

OBJECTIVETo investigate the effects of activation of the GLP-1 receptor on the p38MAPK signaling pathway in hepatic stellate cells (HSCs).

METHODSHSCs were isolated and identified according to morphological features; the levels of GLP-1R protein were determined by western blotting.The HSCs were randomly divided into a control grouP (normal saline treatment) and experimental grouP(liraglutide treatment); after 120 hours, the expression of p38MAPK mRNA was examined by RT-PCR and of phosphorylated (p)-p38MAPK protein was detected by western blotting.

RESULTSGLP-1R proteins were detected in the HSCs. Compared with the control group, the experimental group showed significantly decreased p38MAPK mRNA and p-p38MAPK protein (both P < 0.01).

CONCLUSIONThe p38MAPK signaling pathway could be down-regulated when GLP-1R is activated in HSCs.

Cells, Cultured ; Glucagon-Like Peptide 1 ; analogs & derivatives ; pharmacology ; Glucagon-Like Peptide-1 Receptor ; Hepatic Stellate Cells ; metabolism ; Humans ; Liraglutide ; MAP Kinase Signaling System ; RNA, Messenger ; Receptors, Glucagon ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism