2.Expression of stomatin like protein-2 in gastric adenocarcinoma and its effects on the migration and invasion of SGC7901 cell
Jian ZHANG ; Jiansheng LI ; Min WU ; Liqun ZHAO ; Zisen ZHANG ; Ye ZHAO
Chinese Journal of Digestion 2013;(1):37-41
Objective To investigate the expression of stomatin like protein 2 (SLP-2) in gastric adenocarcinoma and its effects on migration and invasion of SGC7901 cell line.Methods The expression of SLP-2 in 60 pairs of gastric adenocarcinoma tissue specimens and corresponding adjacent tumor tissue specimens was detected by immunohistochemistry,and its relation with clinical pathological characteristics was also analyzed.SGC7901 cells were transient transfected with SLP-2 siRNA.After transfection,the expression of SLP-2 in SGC7901 cells at mRNA and protein level was detected by reverse transcripton-polynerase chain reaction (RT-PCR) and Western blot.The changes of migration and invasion of SGC7901 cells after transfection were observed through Transwell test and Matrigel in vitro invasion test.The data were analyzed by x2 test and LSD-t test.Results The high expression percentage of SLP-2 in gastric adenocarcinoma tissue was 68.3% (41/60),which was significantly higher than that of the adjacent tumor tissues (25.0%,15/60; x2 =22.634,P<0.01).The expression of SLP-2 in late stage gastric adenocarcinoma tissues was higher than that of early stage gastric adenocarcinoma tissues (x2 =4.962,P<0.05).The expression of SLP-2 in gastric adenocarcinoma tissues with lymph nodes metastasis was higher than that in gastric adenocarcinoma tissues without lymph nodes metastasis (x2 =3.994,P<0.05).There was no correlation between the expression and gender,age,tumor size and differentiation degree (all P>0.05).After SLP-2 siRNA transfection,the expression of SLP-2 in SGC7901 cells significantly decreased at both mRNA and protein level.At 48 h after SLP-2 siRNA transfection,the cell migration ability was significantly lower (migrated cell number 53.0± 10.8) than that of negative control group (75.8±13.4) and blank control group (78.4± 15.2) (t=22.778 and 25.444,both P<0.01).The in vitro invasion ability of SGC7901 cells (migrated cell number 36.3 ± 8.9) was also significantly lower than that of negative control group (53.9±12.1) and blank control group (49.8±9.0) (t=17.556 and 13.444,both P<0.01).Conclusions The expression of SLP-2 was high in gastric adenocarcinoma tissues.SLP-2 may involve in the metastasis of gastric adenocarcinoma through promoting the migration and invasion ability of gastric adenocarcinoma cell.
3.Pathophysiological features of rat hemorrhagic shock under high temperature conditions
Sheng MA ; Chenyang DUAN ; Zisen ZHANG ; Hongliang ZHAO ; Tao LI ; Liangming LIU
Chinese Journal of Trauma 2016;32(12):1125-1129
Objective To investigate the pathophysiological features of rat hemorrhagic shock under high temperature conditions.Methods A total of 128 SD rats were assigned to high temperature group and normal temperature group according to the random number table,with 64 rats per group.Rats in high temperature group were pretreated at 342 for 12 h,and in normal temperature group were kept at 25℃.Hemorrhagic shock models in rats were produced by withdrawing 40% of the total blood volume via the femoral artery.Parameters of the two groups were measured including blood electrolytes (Na +,K +),plasma osmotic pressure,liver function [aspartate aminotransferase (AST),alanine aminotransferase (ALT)],kidney function [creatinine (CREA)],arterial blood gases (pH,PO2 and PCO2) and cardiac function [heart rate (HR),cardiac output (CO),cardiac index (CI) and DO2].Animal survival time and survival rate were detected.Results Before shock,high temperature group versus normal temperature group showed higher detections of Na+ concentration [(142.3 ± 2.2) mmol/L:(139.1 ±1.5) mmol/L],plasma osmotic pressure [(304.8 ± 4.7) mmol/L:(300.0 ± 1.9) mmol/L],HR [(462 ±30) times/min:(402 ± 44) times/min],CO [(0.892 ± 0.190) L/min:(0.713 ± 0.090) L/min] and CI [(0.0030±0.0006)L·min-1 · cm-2:(0.0023 ±0.0002)L· min-1 · cm-2] (P<0.05).After shock,normal temperature group showed further increased concentrations of Na + and K + and significantly enhanced AST,ALT and CREA compared to normal temperature group (P < 0.05).After shock,CO,CI and DO2 in high temperature group were decreased by 63%,63% and 69% respectively compared to those before shock,but less decrease was observed in normal temperature group.Rat survival rate and survival time were significantly reduced in high temperature group compared to normal temperature group (P < 0.05).Conclusion Pathophysiological changes of hemorrhagic shock in rats under high temperature are mainly manifested as impaired cardiac function,significantly increased concentrations of Na + and K + and significantly shortened survival time.
4. Effect of miR-513a-3p targeting MDM2 on proliferation, migration and invasion of gastric cancer cells
Yingguang MA ; Zisen ZHANG ; Yong YU ; Xiaofang XU ; Li YUAN
Chinese Journal of Oncology 2020;42(1):30-36
Objective:
To investigate the effects of miR-513a-3p on proliferation, migration and invasion of gastric cancer cells and its mechanism.
Methods:
The miR-NC (miR-negative control mimics), miR-513a-3p (miR-513a-3p mimics), anti-miR-NC, anti-miR-513a-3p, si-NC, si-MDM2 (murine double minute 2), miR-513a-3p+ pcDNA3.1 (co-transfected with miR-513a-3p and pcDNA3.1), miR-513a-3p+ pcDNA3.1-MDM2 (co-transfected with miR-513a-3p and pcDNA3.1-MDM2) were transfected into BGC-823 cells, respectively. The expression of miR-513a-3p was detected by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR), and the protein expressions of cyclin D1, MMP-2, p21, E-cadherin, MDM2 were detected by western blot. The viability of BGC-823 cells of each group was detected by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) assay. The migration and invasion of each group were detected by Transwell, the targeting relationship between miR-513a-3p and MDM2 was detected by double luciferase reporter gene assay.
Results:
The expression of miR-513a-3p in gastric epithelial cells GES-1 was 0.76±0.08, significantly higher than 0.21±0.02 in gastric cancer cells BGC-823 and 0.34±0.03 in MGC-803, respectively (
5.Study on protective effects and mechanism of ziyuglycoside Ⅰ on acute lung injury in sepsis rats
Daiqin BAO ; Yiyan LIU ; Zisen ZHANG ; Han SHE ; Lei TAN ; Tao LI ; Qingxiang MAO ; Liangming LIU
China Pharmacy 2023;34(5):537-543
OBJECTIVE To investigate the protective effects and mechanism of ziyuglycoside Ⅰ on acute lung injury in sepsis rats based on network pharmacology, and conduct experimental verification. METHODS The network pharmacology was used to predict the potential target of ziyuglycoside Ⅰ in the treatment of acute lung injury following sepsis. The rat model of sepsis was reproduced by cecum ligation and puncture for experimental verification. Totally 192 SD rats were randomly divided into the sham operation group (Sham group), sepsis group (Sep group), conventional therapy group (CT group) and ziyuglycoside Ⅰ group (Zg Ⅰ group), respectively. Sham group and Sep group were given sterile normal saline, and CT group and ZgⅠ group were given relevant volume of Ringer’s solution and ziyuglycoside Ⅰ. The arterial blood gas, serum inflammatory factors, lung wet/dry mass ratio, pathological changes of lung tissue, pulmonary vascular permeability, the expressions of pulmonary vein tight junction protein 1 (ZO-1) and vascular endothelial cadherin (VE-cadherin) protein and 72-hour survival were observed in each group. RESULTS Results of network pharmacology showed that there were 47 potential targets of ziyuglycoside Ⅰ in the treatment of sepsis. The results of gene ontology function enrichment analysis and Kyoto encyclopedia of genes and genomes pathway enrichment analysis showed that the mechanism could 598486924@qq.com be correlated with biological processes such as positive regulation of reactive oxygen species metabolism, wound healing, regulation of endothelial cell proliferation, cell activation, blood vessel development, response to oxidative stress, etc., and with signaling pathway such as apoptosis, tight junction, HIF-1 signaling pathway, etc. The results of experimental verification showed that compared with Sham group, pH value and the level of partial arterial oxygen pressure were decreased significantly in Sep group (P<0.05), while the level of partial pressure of carbon dioxide, serum levels of tumor necrosis factor α, interleukin 6 were increased significantly (P<0.05); the ratio of lung wet/dry mass was increased significantly (P<0.05); the protein expressions of ZO-1 and VE-cadherin were decreased significantly (P<0.05); 72 h survival rate decreased,the survival time was significantly shortened (P<0.05); the results of pathological observation of lung tissue showed that the rats’ alveoli were extensively ruptured, the alveolar wall was thickened and accompanied with edema, and there was obvious inflammatory cell infiltration; the results of pulmonary vascular permeability observation showed that the lung surface of rats was dark, with a large amount of Evans blue exudation, and the left lower lung was obviously dark blue. Compared with Sep group, the levels of above indexes almost were reversed significantly in CT group and ZgⅠ group (P<0.05); the lung histopathology and pulmonary vascular permeability were significantly improved, and the recovery degree of ZgⅠ group was greater than that of CT group, which was close to the results of Sham group. CONCLUSIONS Ziyuglycoside Ⅰ can significantly reduce inflammatory reaction and acute lung injury in septic rats, which is related to vascular function and tight junction signal pathway.