Objective To explore the expression of microRNA(miR)-100-5p in thyroid cancer cells and its regulatory effects on cell proliferation and apoptosis through experiments.Methods The relative expressions of miR-100-5p in thyroid cancer cell lines(TPC-1 and KTC-1)and normal thyroid cell lines(Nthy ori3-1)were detected using fluorescence quantitative PCR.After transfection of miR-100-5p mimic,miR-100-5p inhibitor,and corresponding negative controls(miR-mimic NC,miR-inhibitor NC)into TPC-1 cells,the proliferation condition of TPC-1 cells was detected using CCK-8,and the apoptosis condition of TPC-1 cells was detected using flow cytometry.Prediction and functional enrichment analysis of target genes of miR-100-5p were performed using the miRTarBase and TargetScan7.2 databases.The targeted regulatory effect of miR-100-5p on fibroblast growth factor receptor 3(FGFR3)was validated using Western blot and dual luciferase reporter gene experiments.Results Compared with Nthy-ori3-1 cells,the expression levels of miR-100-5p in TPC-1 cells(1.87±0.03 vs 1.00±0.03)and KTC-1 cells(6.33±0.47 vs 1.00±0.03)were both up-regulated,with significant differences(t=-34.220,-19.588,all P＜0.05).The 450nm absorbance(A450nm)of cells transfected with miR-100-5p mimic at 24,48 and 72 h were higher than the miR-mimic NC group,with significant differences(t=-7.516,-17.828,-8.445,all P＜0.05).Conversely,the A450nm values of cells transfected with miR-100-5p inhibitor at 24,48 and 72 h were lower than the miR-inhibitor NC group,with significant differences(t=6.720,6.782,6.073,all P＜0.05).The apoptosis rate after transfection with miR-100-5p mimic was decreased compared to miR-mimic NC group(7.43％±0.49％vs 10.55％±0.80％),with significant differences(t=5.767,P=0.004).Compared to miR-inhibitor NC group,the apoptosis rate after transfection with miR-100-5p inhibitor was increased(3.19％±0.22％vs 2.64％±0.15％),with significant differences(t=-3.606,P=0.023).Western blot experiments showed that FGFR3 protein expression levels in the miR-100-5p mimic group were down-regulated compared to the miR-mimic NC group(0.78±0.12 vs 1.00±0.00),with significant differences(t=3.071,P=0.037).Compared to the miR-inhibitor NC group,FGFR3 protein expression levels in the miR-100-5p inhibitor group were up-regulated(1.17±0.07 vs 1.00±0.00),with significant differences(t=-4.509,P=0.046).There was no significant difference in the luciferase activity of the FGFR3 wild-type(1.01±0.17 vs 1.00±0.00)and mutant groups(0.99±0.11 vs 1.00±0.00)between miR-100-5p mimic and miR-mimic NC,and the differences were statistically significant(f=-0.057,0.181,P=0.96,0.873).Conclusion MiR-100-5p in thyroid cancer cells was up-regulated,which may promote cell proliferation and inhibit apoptosis.It may become a new biomarker and regulatory target in the diagnosis and treatment of thyroid cancer.

Objective To observe the clinical effects of Zaobo decoction combined bisoprolol treating premature ventricular contractions.Methods 108 patients of ventricular premature beat were recruited intoa control group and an observation group (n=54) according to the random number table method.The control group was treated with bisoprolol, while the observation group was treated with Zaobo decoction on the basis of the control group, and both groups were treated for 8 weeks.24 h dynamic electrocardiogram (ECG), renin activity plasma (PRA), angiotensin Ⅱ (angiotensionⅡ) and ALD (Ang) were observed before and after treatment.The clinical effects were evaluated.Results The total effective rate showed significant difference between the observation group and the control group (75.9％ vs.57.4％;x2=4.167, P=0.041) after the treatment.After treatment, Ang-Ⅱ (56.22 ± 12.7 pg/ml vs.68.45 ± 12.7 pg/ml, t=5.004) in the observation group was significantly lower than the control group (P＜0.01);24 h sinus RR interval standard deviation (129.16 ± 28.56 ms vs.116.13 ± 17.38 ms, t=2.864), every 5 min sinus RR interval mean standard deviation within 24 h (123.57 ± 25.24 ms vs.112.46 ± 18.23 ms, t=2.622), and within 24 h of sinus RR interval difference rms (31.76 ± 11.42 ms vs.22.64 ± 10.32 ms, t=4.354) in the observation group were significantly higher than the control group (P＜0.01).Conclusion Zaobo decoction combined with bisoprolol can effectively improve heart rate variability, regulate rernin vascular angiotensin system, and improve the clinical efficacy of the patients with ventricular premature beat.

OBJECTIVETo detect the expressions of CXCR4 and Nrf2 in non-small cell lung cancer (NSCLC) tissues and analyze their association with the clinicopathological features of NSCLC.

METHODSWe investigated the expressions of CXCR4 and Nrf2 in 66 NSCLC and corresponding distant normal tissue specimens using immunohistochemistry and real-time PCR.

RESULTSThe expressions of CXCR4 protein and mRNA were significantly higher in NSCLC tissue specimens than in the distant normal tissues, while the expression of Nrf2 protein and mRNA increased significantly in NSCLC tissues compared to those in the distant normal tissues (P<0.01). A high expression level of CXCR4 was positively correlated with a large tumor size (P=0.048), poor differentiation (P=0.024), advanced TNM stage (P=0.018), lymph node metastasis (P=0.004), and distant metastasis (P=0.016). The expression of Nrf2 protein was positively correlated with a large tumor size (P=0.008), advanced TNM stage (P=0.028), lymph node metastasis (P=0.038), and distant metastasis (P=0.023). A strong correlation was found between CXCR4 and Nrf2 expressions in NSCLC tissues (r=0.324, P<0.01), and the co-expression of CXCR4 and Nrf2 was strongly correlated with lymph node metastasis and distant metastasis.

CONCLUSIONAbnormal expressions of CXCR4 and Nrf2 may contribute to the progression and malignant biological behavior of NSCLC.

Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; NF-E2-Related Factor 2 ; metabolism ; Neoplasm Staging ; Receptors, CXCR4 ; metabolism ; Signal Transduction

OBJECTIVETo observe autophagy induced by starvation in non-small cell lung cancer A459 and 95D cells.

METHODSA549 and 95D cells in logarithmic growth in 1640 medium were cultured in Earle's balanced salt solution (EBSS) for 0, 1, 2, 3, 4 or 5 h. Autophagosome formation in the cell culture was observed by MDC fluorescent staining, and the expression of microtubule-associated protein 1 light chain 3 (LC3) and Beclin1 in the cells were detected using Western blotting.

RESULTSCompared with the control cells, the cells with prolonged starvation showed increased MDC-positive cells and autophagosome formation. The expression of Beclin-1 and the LC3-II/LC3-I ratio also increased as the starvation prolonged, reaching the peak levels at 3 h and 4 h, respectively.

CONCLUSIONAutophagy can be induced by starvation in A549 and 95D cells in correlation with the expression of autophagy-related proteins LC3 and Beclin-1. These cell models of nutritional deficiency-induced autophagy may allow for a better understanding of the role of autophagy in the development of non-small cell lung cancer.

Apoptosis Regulatory Proteins ; metabolism ; Autophagy ; Beclin-1 ; Carcinoma, Non-Small-Cell Lung ; pathology ; Cell Line, Tumor ; Humans ; Membrane Proteins ; metabolism ; Microtubule-Associated Proteins ; metabolism

Objective To explore the evolution of research themes and trends in China's nursing college education during the last decade.Methods A co-word analysis of keywords was performed in the research literatures around nursing college education from two Chinese databases,VIP China Science and Technology Journal Database and CKNI Periodical Full-text Database,between 2003 and 2012.The analysis was based on two different periods (phase Ⅰ from 2003 to 2007 and phase Ⅱ from 2008 to 2012).Results A total of 646 articles were included,with 287 articles published in phase Ⅰ and 359 articles in phase Ⅱ.Sixteen high-frequency keywords were identified during the phase Ⅰ,with seventeen during the phase Ⅱ.Two research themes,nursing student-training model and construction and reform of curriculum,were presented in both phases.However,some differences in research sub-themes exited between phase Ⅰ and phase Ⅱ.Conclusions Hot research topics in nursing college education seemed no change in the past ten years.School-hospital collaboration newly emerged as a hot topic in the field of nurse education research.The research on construction and reform of curriculum change its focus to nursing courses and humanities courses.The academic studies of nursing college education were mostly influenced and pushed by reforming and developing of nursing education.

Objective To detect the expressions of CXCR4 and Nrf2 in non-small cell lung cancer (NSCLC) tissues and analyze their association with the clinicopathological features of NSCLC. Methods We investigated the expressions of CXCR4 and Nrf2 in 66 NSCLC and corresponding distant normal tissue specimens using immumohistochemistry and real-time PCR. Results The expressions of CXCR4 protein and mRNA were significantly higher in NSCLC tissue specimens than in the distant normal tissues, while the expression of Nrf2 protein and mRNA increased significantly in NSCLC tissues compared to those in the distant normal tissues (P<0.01). A high expression level of CXCR4 was positively correlated with a large tumor size (P=0.048), poor differentiation (P=0.024), advanced TNM stage (P=0.018), lymph node metastasis (P=0.004), and distant metastasis (P=0.016). The expression of Nrf2 protein was positively correlated with a large tumor size (P=0.008), advanced TNM stage (P=0.028), lymph node metastasis (P=0.038), and distant metastasis (P=0.023). A strong correlation was found between CXCR4 and Nrf2 expressions in NSCLC tissues (r=0.324, P<0.01), and the co-expression of CXCR4 and Nrf2 was strongly correlated with lymph node metastasis and distant metastasis. Conclusion Abnormal expressions of CXCR4 and Nrf2 may contribute to the progression and malignant biological behavior of NSCLC.

Objective To observe autophagy induced by starvation in non-small cell lung cancer A459 and 95D cells. Methods A549 and 95D cells in logarithmic growth in 1640 medium were cultured in Earle's balanced salt solution (EBSS) for 0, 1, 2, 3, 4 or 5 h. Autophagosome formation in the cell culture was observed by MDC fluorescent staining, and the expression of microtubule-associated protein 1 light chain 3 (LC3) and Beclin1 in the cells were detected using Western blotting. Results Compared with the control cells, the cells with prolonged starvation showed increased MDC- positive cells and autophagosome formation. The expression of Beclin-1 and the LC3-II/LC3-I ratio also increased as the starvation prolonged, reaching the peak levels at 3 h and 4 h, respectively. Conclusion Autophagy can be induced by starvation in A549 and 95D cells in correlation with the expression of autophagy-related proteins LC3 and Beclin-1. These cell models of nutritional deficiency-induced autophagy may allow for a better understanding of the role of autophagy in the development of non-small cell lung cancer.

Objective To detect the expressions of CXCR4 and Nrf2 in non-small cell lung cancer (NSCLC) tissues and analyze their association with the clinicopathological features of NSCLC. Methods We investigated the expressions of CXCR4 and Nrf2 in 66 NSCLC and corresponding distant normal tissue specimens using immumohistochemistry and real-time PCR. Results The expressions of CXCR4 protein and mRNA were significantly higher in NSCLC tissue specimens than in the distant normal tissues, while the expression of Nrf2 protein and mRNA increased significantly in NSCLC tissues compared to those in the distant normal tissues (P<0.01). A high expression level of CXCR4 was positively correlated with a large tumor size (P=0.048), poor differentiation (P=0.024), advanced TNM stage (P=0.018), lymph node metastasis (P=0.004), and distant metastasis (P=0.016). The expression of Nrf2 protein was positively correlated with a large tumor size (P=0.008), advanced TNM stage (P=0.028), lymph node metastasis (P=0.038), and distant metastasis (P=0.023). A strong correlation was found between CXCR4 and Nrf2 expressions in NSCLC tissues (r=0.324, P<0.01), and the co-expression of CXCR4 and Nrf2 was strongly correlated with lymph node metastasis and distant metastasis. Conclusion Abnormal expressions of CXCR4 and Nrf2 may contribute to the progression and malignant biological behavior of NSCLC.

Objective To observe autophagy induced by starvation in non-small cell lung cancer A459 and 95D cells. Methods A549 and 95D cells in logarithmic growth in 1640 medium were cultured in Earle's balanced salt solution (EBSS) for 0, 1, 2, 3, 4 or 5 h. Autophagosome formation in the cell culture was observed by MDC fluorescent staining, and the expression of microtubule-associated protein 1 light chain 3 (LC3) and Beclin1 in the cells were detected using Western blotting. Results Compared with the control cells, the cells with prolonged starvation showed increased MDC- positive cells and autophagosome formation. The expression of Beclin-1 and the LC3-II/LC3-I ratio also increased as the starvation prolonged, reaching the peak levels at 3 h and 4 h, respectively. Conclusion Autophagy can be induced by starvation in A549 and 95D cells in correlation with the expression of autophagy-related proteins LC3 and Beclin-1. These cell models of nutritional deficiency-induced autophagy may allow for a better understanding of the role of autophagy in the development of non-small cell lung cancer.

Objective Amniotic membrane as the carrier to culture the vascular endothelial cells was investigated in this study in order to explore whether the corneal endothelial cells superseded by the vascular endothelial cells is feasible on the account of directing a kind of the original method to handle the bullous keratopathy.Methods The vascular endothelial cells adhering to the sphagitides lumen of the experimental rabbits were digested and gained by means of the perfusion method with 0.25%tripsin plus 0.02%EDTA.Fresh amniotic membrane without the amniotic epithelial cells was cut into the square tissue about 1.5 cm?1.5 cm and spread evenly in the 24-well culture dish.Primary cultured cells were subcultured on the amniotic membrane.We would not transplant the vascular endothelial cells feeding on the amniotic membrane until cells is full of the whole amniotic membrane surface.One to two months after the operation,the change of corneal diaphaneity was observed.Results About 12 days since the cell transplantation was performed,the corneal transparency alteration between the experimental groups and the control one is different.The corneal buttons in the experimental group show the severe edema and opacity,and the anterior chamber couldn't be seen unclearly.But,10 days after the operation,the corneal oedema which begins to extenuate was judged through the indicatrix that the corneal edema in the experimental group has been recovering slowly,among which the anterior chamber tissue of 7 animals was visible through the implant.The corneal edema in the control groups intensified evidently,even,the part of these appeared the ulcerous necrosis as result of the corneal severely edema.There is the existence of difference between two groups(P