To investigate the effects of Snail1 gene silence on the expression of tight junction proteins and the migration ability of Hep-2 cells, Hep-2 cells were transfected with plasmids which is containing the shRNA of Snail1 gene, and cultured till the cells could be passaged stably (named Sh-snail1 cells). The expression of tight junction proteins (ZO-1, Occludin, Claudin-5) were detected by Western blot. The migration ability of Sh-snail1 cells was investigated by wound healing assay, and the protein expression of members of RhoGTPase family (RhoA, Cdc42) was detected by Western blot, which is closely related to the migration ability. Our results showed that the expression of tight junction proteins (ZO-1, Occludin, Claudin-5) was significantly increased; the migration ability of Sh-snail1 cell was inhibited; the expression of RhoA and Cdc42 was downregulated. All of these indicated that silencing the gene of Snail1 in Hep-2 cells can up-regulate the expression of tight junction proteins and down regulate the expression of Cdc42 and RhoA, and further inhibit the migration of Hep-2 cells. Furthermore, opening of the tight junctions between cells and the stronger migration ability of cancer cells are important processes in cancer metastasis. It is confirmed that the Snail1 gene is closely related to the two processes, providing an experimental basis for targeted therapy of laryngeal squamous cell carcinoma.
Cell autophagy plays a key role in maintaining intracellular nutritional homeostasis during starvation through elimination of aberrant or obsolete cellular structures. The cellular cytoskeleton has a crucial role in multiple processes involving membrane rearrangements and vesicle-mediated events. Autophagy is mediated by both microtubules and actin networks: microtubules promote the synthesis of autophagosome and are related to the movement of autophagosome; actin networks have been implicated in structurally supporting the expanding of phagophore, moving autophagosomes and enabling their efficient fusion with the lysosome; non-muscle myosinⅡoperates in the early stages of autophagy during the initiation and expansion of the phagophore, whereas myosinⅥ and myosin 1C are involved in the late stages of autophagosome maturation and fusion with the lysosome, respectively. This review summarizes the multiple regulation of cytoskeleton on autophagy and focuses on the regulation of autophagy by actin and myosin, providing a new approach for the study of pathogenesis and innovative therapies of autophagy related diseases.
ObjectiveTo find out the influencing factors of electrocardiogram (ECG) monitor configuration decision in surgical nursing units and form a scientific configuration standard, so as to provide a basis for the reasonable configuration of ECG monitors.MethodsFrom May to June 2018, the indexes and weights affecting the configuration of ECG monitors in surgical nursing units of a large public hospital were determined by interview survey method and analytic hierarchy process.ResultsThe influencing factors for configuration of ECG monitors in surgical nursing units were the number of operations, number of rescues, number of emergencies, number of deaths, and number of patients transferred to and out of intensive care unit, and the weights were 0.459 7, 0.224 9, 0.155 3, 0.111 2, and 0.049 0, respectively. The classification of nursing units was taken as plan, and the configuration standard of ECG monitors was established.ConclusionThe configuration model of ECG monitors in surgical nursing units based on analytic hierarchy process realizes the combination of qualitative and quantitative analysis, which provides scientific and reasonable reference for the configuration of ECG monitors.
Objective To explore the pathogenesis of the level of gene and therapeutic target genes associated with intestinal obstruction by analyzing the differential expression gene. Methods The gene expression data that came from public database gene expression omnibus （GEO） which provided adhesion formation’ gene expression data on 1， 3， 7，and 14 days after operation （n=8） and normal intestinal tissues’ gene expression data （n=2） of mouse were collected. The gene function and differential expression of genes were analyzed by using gene ontology （GO） and significance analysis of microarray （SAM）. Results There were a lot of response stimulated up-regulation of gene expression when occurrence of adhesion， and the products of these genes were distributed on cell membrane. The analysis results of gene expression at different time point after operation showed that expression up-regulated of Hmgcs 2 gene occurred on 3-14 days ofter operation and expression up-regulated of Stxbp 5 gene occurred on 14 days ofter operation. Conclusions The adhesion formation may be closely associated with the genes of response to stimulus and the gene product in membrane. The Hmgcs 2 and Stxbp 5 genes may be closely associated with the occurrence of other diseases which induced by adhesion formation.This provides a basis for the discovery of potential therapeutic targets.
Objective To study oxygen carrying capacity of red blood cell in children with congenital left-to-right shunt (CLRS) and discuss its clinical significance. Methods A total of 62 children with CLRS were selected as a trial group and 40 healthy children who had accepted clinical physical examination as a control group. ELISA test was applied to determine 2,3 -DPG content of red blood cells. At the same time, pH, PaO2, PaCO2, Hb, P50O2 and relevant outcomes were tested for correlation analysis. Results Red blood cell 2,3-DPG (2.76±0.98 μmol/mL) was higher than that of the control group. PaO2, pH, Hb were lower than those of the control group with significant differences. Among the types of CLRS, 2,3 -DPG of ventricular septal defect was the highest and that of patent ductus arteriosus ranked the second, the two of which had a significant difference (P=0.007). As for red blood cell of children with congenital heart diseases, there was linear correlation between 2,3- DPG and PaO2 as [2,3-DPG=12.007 8– (0.154 7×PaO2)], as well as between P50O2 and 2,3 -DPG as [P50O2=26.303 6+ (1.799 2×2,3-DPG)]. Conclusion Children with congenital left-to-right shunt tend to have low oxygen. Therefore, it is important to detect 2,3-DPG of red blood cell, blood gas analysis, and hemoglobin level change, in order to well understand the mechanism of congenital heart disease as well as to guide clinical practice.
The article aims to explore the optimal concentration of arsenic trioxide (As2O3) on HepG2 of liver cancer cells, and the effect of As2O3 on the migration, invasion and apoptosis of HepG2 cells. In this study, the activity of HepG2 cells treated with 0, 1, 2, 4, 8, 16, 32 μmol/L As2O3 was tested by CCK-8 method, the semi-inhibitory concentration (IC50) was calculated, and the morphological changes of HepG2 cells were observed after the action of As2O3 at IC50 concentration for 12, 24, 48 h. The effect of As2O3 on cell migration and invasion ability was verified by wound healing experiment and Transwell invasion experiment. Western blot and qRT-PCR were used to detect the effects of As2O3 on the gene and protein expression levels related to cell migration, invasion and apoptosis. The results showed that, compared with the control group, the activity of HepG2 cells decreased with the increase of the concentration of As2O3 treatment, showing a dose-dependent effect, and its IC50 was 7.3 μmol/L. After 24 hours’ treatment with 8 μmol/L As2O3, HepG2 cells underwent significant apoptosis, and its migration and invasion abilities were significantly reduced. In addition, the protein expression levels of RhoA, Cdc42, Rac1 and matrix metalloproteinase-9 (MMP-9) were down-regulated, the protein and mRNA expression levels of anti-apoptotic gene Bcl-2 were significantly down-regulated, and the protein and mRNA expression levels of pro-apoptotic genes Bax and Caspase-3 were significantly up-regulated. The above results indicate that certain concentration of As2O3 can inhibit the migration and invasion of hepatocellular carcinoma cells and promote the apoptosis of hepatocellular carcinoma cells.
This study aims to investigate the effect of substances secreted or metabolized by vascular endothelial cells on epithelial-mesenchymal transition (EMT) of hepatocellular carcinoma cells under indirect co-culture condition. Human hepatocellular carcinoma cell line QGY-7703 was cultured in vitro, and then was co-cultured with conditioned medium of human umbilical vein endothelial cells (HUVEC). The morphological changes of QGY-7703 cells were observed by inverted phase contrast microscopy. The migration ability of QGY-7703 cells was analyzed by scratch-wound assays. The effect of conditioned medium on the expression and distribution of EMT related proteins was detected by Western blot and immunofluorescence assays, respectively. The results showed that the QGY-7703 cells gradually changed from polygonal to spindle shape, the migration ability promoted significantly, and both the expression and distribution of EMT related marker changed in a time-dependent manner after co-culturing. The results confirm that vascular endothelial cells can induce EMT in hepatocellular carcinoma cells under indirect co-culture condition.