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Purpose To investigate the expression of miR200a in different lung cancer cell lines and its effect on proliferation,migration,and apoptosis in A549 cells.Methods The expressions of miR-200a in different lung cancer cell lines were detected by RT-PCR.miR-200a mimics was transfected into A549 cells by Lipofectamine RNAiMax.The change of proliferation ablility of A549 cells was detected by CCK-8 method and plate clone formation assay.Cell migration was examined by Transwell chamber assay.The flow cytometry was used to examine the changes of apoptosis.The possible target genes of miR-200a were forecasted by bioinformatics tools.Results The results of RT-PCR showed that the expression of miR-200a was significantly down-regulated in A549,H23 and H460 cell lines than 16HBE cell line.CCK-8 assay showed that the OD values of the mimics group at 4,and 5 days were significantly lower than those in the negative control (NC) group (P ＜ 0.05).Plate clone formation assay showed rate of colony formation in the mimics group was significantly lower than that in the NC group [(33.13±0.74)％ vs (45.57 ±1.27)％,P＜0.05].Transwell migration assay showed that the cell number of mimics group that passed the Transwell membrane was significantly lower than that of the NC group [(71.60 ± 17.90) vs (140.20 ± 17.88),P ＜0.05].Flow cytometry showed that the apoptosis rate of the mimics group was significantly higher than that of the NC group [(17.80± 1.90)％ vs (11.33 ± 1.96)％,P ＜ 0.05].Tiam1 may be one of the target gene of miR-200a.Conclusion miR-200a can inhibit the proliferation and migration,and promote apoptosis of lung cancer A549 cells,suggesting a potential new therapeutic agent for lung cancer cell.MiR-200a may play a function of regulation of tumor development through target gene Tiam1.
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Objective To investigate awareness and attitude of advance directives among ICU nurses and analyze its influencing factors,and to provide references for promotion and application of advance directives in China.Methods A questionnaire survey of 256 ICU nurses from secondary and higher hospitals in Tianjin was conducted from December 2016 to January 2017,and a self-designed questionnaire of awareness and attitude of advance directives was used to measure nurses' awareness,attitude and obstacles of promotion.Results The total score of advance directives knowledge was(23.45±6.34).There were 222 nurses(90.6％)who supported the use of advance directives.‘Our country has no relevant legal support,and it is difficult to implement advance directives’ was the primary obstacle.The number of cared patients daily,children situation and hospital level were influencing factors of attitude (P＜0.05).Conclusion ICU nurses have relatively low level of knowledge of advance directives.ICU nurses hold a positive attitude towards advance directives,but there are still concerns;nurses who have the number of cared patients daily ≥3,have children,and from tertiary hospitals have a higher rate of approval.
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Objective To observe the effect of electroacupuncture(EA)at Baihui(GV20)and Siguan(Hegu/LI4 and Taichong/LR3)of affected side on expression of Tax1-binding protein 1(TAX1BP1)in cerebral cortex in focal cerebral ischemia-reperfusion rats,so as to in-vestigate its protective mechanism in inhibiting nuclear factor kappa B(NF-κB)signaling pathway and promoting neurobehavioral recovery. Methods A total of 105 Sprague-Dawley rats were randomly assigned into sham group,model group and EA group.Each group was ran-domly assigned into reperfusion six hours,twelve hours,24 hours,48 hours,72 hours groups after two hours of ischemia.The model was es-tablished by right middle cerebral artery occlusion and reperfusion.EA group received electroacupuncture at Baihui and left Siguan(Hegu and Taichong)acupoints.Neurobehavioral evaluation,TAX1BP1 protein expression,TAX1BP1 positive cell count,zink finger protein A20 expression, and nuclear NF-κB p65 protein expression were tested in each group. Results There was no neurological deficit in the sham group. Compared with the model group, the neurological scores at 48 hours, 72 hours after reperfusion decreased in EA group (P<0.05). Compared with the control group,the TAX1BP1 expression at twelve hours,24 hours and 48 hours after reperfusion increased in the model group(P<0.05),and further increased at twelve hours,24 hours,48 hours and 72 hours after reperfusion in EA group(P<0.05),and peaked at 24 hours after reperfusion.Compared with the sham group,the expression of A20 and NF-κB p65,and the number of TAX1BP1 positive cells increased in the model group(P<0.05),and the expression of A20 and the number of TAX1BP1 positive cells further increased,(P<0.05)and the expression of NF-κB p65 decreased in EA group(P<0.05)at 24 hours after reperfusion.Immunofluorescence labeling indicat-ed that TAX1BP1 protein primarily expressed in the cytoplasm,TAX1BP1 protein and A20 protein co-expressed in the cytoplasm.Immuno-histochemistry showed indicated that NF-κB p65 mainly expressed in the nucleus in the model groupr,and mainly expressed in the cyto-plasm in the EA group.Conclusion Electroacupuncture could significantly inhibit neuronal NF-κB signaling pathway and promote neurobe-havioral recovery in focal cerebral ischemia-reperfusion,which may related with up-regulating TAX1BP1 protein expression.