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Objective To explore the molecular mechanism of miR-200b-3p regulates the prolifera-tion,invasion,migration and apoptosis of pancreatic cancer cells by targeting vascular endothelial growth factor A (VEGFA). Methods The expression of miR-200b-3p in pancreatic cancer tissues and cell lines was detec-ted by quantitative real-time fluorescence PCR (qRT-PCR). Pancreatic cancer PANC-1 cells were divided into NC group,miR-200b-3p mimic group,si-VEGFA group and si-VEGFA + miR-200b-3p inhibitor group. The proliferation,migration and invasion of PANC-1 cells were measured by CCK-8 and Transwell assay. The apop-tosis of PANC-1 cells was detected by Annexin V-FITC/ PI double staining flow cytometry assay. The targeted relationship of miR-200b-3p and VEGFA was estimated by dual luciferase reporter gene assay and Western blotting. Results The expression of miR-200b-3p in pancreatic cancer tissues and cell lines was decreased. After miR-200b-3p was overexpressed in PANC-1 cells for 48 h,the cell viabilities of PANC-1 cells in NC group and miR-200b-3p mimic group were 1. 250 ± 0. 028 and 0. 983 ± 0. 044,the numbers of migrated cells were 402. 700 ± 21. 530 and 158. 000 ± 17. 620,the numbers of invaded cells were 478. 300 ± 31. 050 and 170. 000 ± 32. 470,and the cell apoptosis rates were (5. 280 ± 0. 352)％ and (7. 430 ± 0. 393)％ . The cell viability,migration and invasion of PANC-1 cells in miR-200b-3p mimic group were significantly decreased than those in NC group (t = 5. 060,P = 0. 007;t = 8. 796,P = 0. 001;t = 6. 863,P = 0. 002). The cell apop-tosis rate in miR-200b-3p mimic group was significantly higher than that in NC group (t = 4. 076,P = 0. 015). The fluorescence intensity in VEGFA-WT group was 1. 000 ± 0. 027,which was significantly higher than that in VEGFA-WT + miR-200b-3p mimic group (0. 632 ± 0. 048;t = 6. 637,P = 0. 003). The fluorescence intensi-ties in VEGFA-MUT group and VEGFA-MUT + miR-200b-3p mimic group were 1. 000 ± 0. 049 and 0. 868 ± 0. 047,with no statistically significant difference (t = 1. 944,P = 0. 124). After miR-200b-3p was overex-pressed in PANC-1 cells for 48 h,the expressions of VEGFA in NC group and miR-200b-3p mimic group were 1. 000 ± 0. 058 and 0. 762 ± 0. 020,respectively. The expression level in miR-200b-3p mimic group was lower than that in NC group (t = 3. 908,P = 0. 017). After transfection of PANC-1 cells with si-VEGFA or si-VEGFA + miR-200b-3p inhibitor for 48 h,the cell viabilities of PANC-1 cells in NC group,si-VEGFA group and si-VEGFA + miR-200b-3p inhibitor group were 1. 300 ± 0. 058,0. 943 ± 0. 047 and 1. 143 ± 0. 023,the numbers of migrated cells were 446. 000 ± 17. 350,206. 300 ± 19. 360 and 428. 300 ± 30. 330,and the numbers of invaded cells were 510. 300 ± 24. 550,175. 700 ± 24. 290 and 473. 700 ± 35. 530,with statisti-cally significant differences (F = 15. 830,P = 0. 004,F = 33. 530,P = 0. 001,F = 38. 860,P < 0. 001). The cell viability,migration and invasion of PANC-1 cells in si-VEGFA group were significantly decreased than those in NC group (P = 0. 003,P < 0. 001,P < 0. 001). There was no significant difference between si-VEGFA + miR-200b-3p inhibitor group and NC group (P = 0. 107,P = 0. 854,P = 0. 671). The cell apop-tosis rates in NC group,si-VEGFA group and si-VEGFA + miR-200b-3p inhibitor group were (3. 810 ± 0. 577)％,(7. 373 ± 0. 482)％ and (3. 650 ± 0. 514)％,with a statistically significant difference (F =16. 020,P = 0. 004). The cell apoptosis rate in si-VEGFA group was significantly higher than that in NC group (P = 0. 007),but there was no significantly difference between si-VEGFA + miR-200b-3p inhibitor group and NC group (P = 0. 975). Conclusion miR-200b-3p suppresses the proliferation,invasion and migration and promotes the apoptosis of pancreatic cells by down-regulating VEGFA.
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Objective@#To explore the molecular mechanism of miR-200b-3p regulates the proliferation, invasion, migration and apoptosis of pancreatic cancer cells by targeting vascular endothelial growth factor A (VEGFA).@*Methods@#The expression of miR-200b-3p in pancreatic cancer tissues and cell lines was detected by quantitative real-time fluorescence PCR (qRT-PCR). Pancreatic cancer PANC-1 cells were divided into NC group, miR-200b-3p mimic group, si-VEGFA group and si-VEGFA+ miR-200b-3p inhibitor group. The proliferation, migration and invasion of PANC-1 cells were measured by CCK-8 and Transwell assay. The apoptosis of PANC-1 cells was detected by Annexin V-FITC/PI double staining flow cytometry assay. The targeted relationship of miR-200b-3p and VEGFA was estimated by dual luciferase reporter gene assay and Western blotting.@*Results@#The expression of miR-200b-3p in pancreatic cancer tissues and cell lines was decreased. After miR-200b-3p was overexpressed in PANC-1 cells for 48 h, the cell viabilities of PANC-1 cells in NC group and miR-200b-3p mimic group were 1.250±0.028 and 0.983±0.044, the numbers of migrated cells were 402.700±21.530 and 158.000±17.620, the numbers of invaded cells were 478.300±31.050 and 170.000±32.470, and the cell apoptosis rates were (5.280±0.352)% and (7.430±0.393)%. The cell viability, migration and invasion of PANC-1 cells in miR-200b-3p mimic group were significantly decreased than those in NC group (t=5.060, P=0.007; t=8.796, P=0.001; t=6.863, P=0.002). The cell apoptosis rate in miR-200b-3p mimic group was significantly higher than that in NC group (t=4.076, P=0.015). The fluorescence intensity in VEGFA-WT group was 1.000±0.027, which was significantly higher than that in VEGFA-WT+ miR-200b-3p mimic group (0.632±0.048; t=6.637, P=0.003). The fluorescence intensities in VEGFA-MUT group and VEGFA-MUT + miR-200b-3p mimic group were 1.000±0.049 and 0.868±0.047, with no statistically significant difference (t=1.944, P=0.124). After miR-200b-3p was overexpressed in PANC-1 cells for 48 h, the expressions of VEGFA in NC group and miR-200b-3p mimic group were 1.000±0.058 and 0.762±0.020, respectively. The expression level in miR-200b-3p mimic group was lower than that in NC group (t=3.908, P=0.017). After transfection of PANC-1 cells with si-VEGFA or si-VEGFA + miR-200b-3p inhibitor for 48 h, the cell viabilities of PANC-1 cells in NC group, si-VEGFA group and si-VEGFA + miR-200b-3p inhibitor group were 1.300±0.058, 0.943±0.047 and 1.143±0.023, the numbers of migrated cells were 446.000±17.350, 206.300±19.360 and 428.300±30.330, and the numbers of invaded cells were 510.300±24.550, 175.700±24.290 and 473.700±35.530, with statistically significant differences (F=15.830, P=0.004, F=33.530, P=0.001, F=38.860, P<0.001). The cell viability, migration and invasion of PANC-1 cells in si-VEGFA group were significantly decreased than those in NC group (P=0.003, P<0.001, P<0.001). There was no significant difference between si-VEGFA + miR-200b-3p inhibitor group and NC group (P=0.107, P=0.854, P=0.671). The cell apoptosis rates in NC group, si-VEGFA group and si-VEGFA+ miR-200b-3p inhibitor group were (3.810±0.577)%, (7.373±0.482)% and (3.650±0.514)%, with a statistically significant difference (F=16.020, P=0.004). The cell apoptosis rate in si-VEGFA group was significantly higher than that in NC group (P=0.007), but there was no significantly difference between si-VEGFA + miR-200b-3p inhibitor group and NC group (P=0.975).@*Conclusion@#miR-200b-3p suppresses the proliferation, invasion and migration and promotes the apoptosis of pancreatic cells by down-regulating VEGFA.
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Objective To study the clinical significance of the expression of D-amino acid oxidase(DAO) in hepatocellular carcinoma(HCC) tissues.Methods The gene expression profiles and related clinical data of 214 HCC cases were collected.Univariate and multivariate analyses were conducted to investigate the association between DAO expression levels,clinical traits,the prognosis.Results Univariate analysis indicated that there were a lower level of blood alpha fetoprotein(AFP,P=0.001),a smaller number of nodules(P=0.042),a better TNM stage(P=0.014),a lower metastasis risk(P=0.001) and a better prognosis(P=0.011)in the samples with high DAO expression.Multivariate analysis also indicated a lower AFP level(OR:0.162,95%CI:0.078-0.336) and metastasis risk(OR:0.140,95%CI:0.069-0.284) in the samples with high DAO expression,as well as a better prognosis(OR:0.833,95%CI:0.700-0.992).Conclusion DAO expression was associated with blood AFP levels,metastasis risk and prognosis in HCC,and its high expression was a protective factor for HCC.
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BACKGROUND:Type 2 diabetic patients suffered from different levels of insulin resistance and insulin secretion defects. Retinol binding protein 4 is a fat derived factor found in the blood and is considered tobe an important target for the treatment of visceral obesity and insulin resistance. OBJECTIVE:To observe the level of serum inflammatory cytokines and the changes of immune function in insulin resistance model rats after swimming. METHODS:A total of28 8-week-old Sprague-Dawleymale ratmodels were randomly divided into swimming group and rest group. Rat received intraperitoneal injection with 3 μg/g recombinant serum retinol binding protein 4, twice a day, with an interval of 12 hours, for 6 weeks. At 6 weeks after administration, rats in the swimming group were subjected to swimming without loading for 6 weeks, 60 minutes per day. Serum retinol binding protein 4 levels were measured with enzyme linked immunosorbent assay (ELISA) kit. Insulin sensitivity (insulin resistance index) was assessed with homeostasis model assessment. Terminal deoxynucleotidyl transferase dUTP nick end labeling method was used to mark the apoptotic cels in rat thymus. ELISA was used to detect serum interleukin-6 expression. ABC-ELISA was used to test Level of serum granulocyte macrophage colony stimulating factor. S-P One Step Method was used to test CD4 and CD8 activities in blood. RESULTS AND CONCLUSION:Interleukin-6 and granulocyte macrophage colony stimulating factor were significantly positively associated with insulin resistance index. The increase in insulin resistance index would induce the increase in the levels of serum inflammatory cytokine interleukin 6 and granulocyte macrophage colony stimulating factor, but exercises could decrease above levels. Thymus index, CD4, and CD4/CD8 were negatively associated with insulin resistance index. These results indicate that exercise intervention effectively improved immunity function of rats with insulin resistance and reduced insulin resistance index.
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Objective To investigate the effect of abdominal vibration therapy on chronic obstructive pulmonary disease (COPD) patients with constipation. Methods In the respiratory department from August 2013 to July 2014, eighty-three COPD patients with constipation were divided into the experiment group (n=35) and the control group (n=48) using random digit table. The control group were treated with conventional western medicine and the experiment group received impingement abdomen vibration therapy on the basis of western medicine treatment. The two groups were compared in terms of symptoms of constipation and curative effect. Results The curative effect of the experiment groups was significantly better than that of the control group (P<0.05). The score on constipation in the experiment group was significantly lower than that of the control group (P<0.05). Conclusion The abdominal vibration therapy is effective in the treatment of COPD patients with constipation. It can improve their quality of life and be worthy of clinical application.
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<p><b>OBJECTIVE</b>To define the determinants of perioperative death and complications after cardiac valve replacement in 702 patients.</p><p><b>METHODS</b>Clinical data of the patients after cardiac valve replacement were analyzed retrospectively.</p><p><b>RESULTS</b>Perioperative mortality and morbidity correlated significantly with some of the perioperative variables, such as higher NYHA functional class (III or IV), large left ventricular end-diastolic diameter (>/= 70 mm), C/T >/= 0.70, prolonged aortic cross-clamping time and cardiopulmonary bypass time, unsatisfactory myocardial protection.</p><p><b>CONCLUSIONS</b>Perioperative mortality and morbidity correlate significantly with some of perioperative variables, such as higher NYHA functional class, unsatisfactory myocardial protection, inappropriate surgical procedure, improper therapy of some complications after cardiac valve replacement. To avoid the occurrence of these independent predictors or to correct them timely might effectively decrease the perioperative mortality and morbidity after cardiac valve replacement.</p>