1.Effect of vitamin E succinate on autophagy in human gastric cancer cells via mitochondria-associated endoplasmic reticulum membranes
Miaomiao CAO ; Fangyu CHEN ; Zhiyang WEI ; Mengmeng LÜ ; Ziqing XING ; Jinze WANG ; Shuang LI ; Liying HOU
Chinese Journal of Pathophysiology 2025;41(11):2157-2165
AIM:This study aims to investigate whether vitamin E succinate(VES)induces autophagy in hu-man gastric cancer cells through the promotion of mitochondria-associated endoplasmic reticulum membranes(MAMs).METHODS:Human gastric cancer cell lines MKN28 and MKN45 were cultured in vitro.Cell viability was assessed us-ing the CCK8 assay,and two cell growth curves were plotted to determine the treatment concentration of VES.Control groups,VES dose groups(MKN28:5,10,20,and 40 mg/L;MKN45:10,20,40,and 80 mg/L),an autophagy-posi-tive control group(rapamycin,RAPA,100 nmol/L),and a MAMs-positive control group(oligomycin A,10 mg/L)were set up.Cells were harvested after 24 h of treatment for subsequent experiments.The formation of autophagosomes and MAMs was observed using transmission electron microscopy.The expression levels of autophagy-related proteins,includ-ing beclin-1,LC3-II/LC3-I,and p62,were detected by Western blot.MAMs labeled with split green fluorescent protein(GFP)were visualized by fluorescence microscopy.The expression of mitofusin 2(MFN2),a key molecule of MAMs,was also detected by Western blot.To inhibit MFN2 specifically,the cells were treated with mitochondrial fusion inhibitor 8(MFI8)and simultaneously transfected with an MFN2 plasmid to achieve MFN2 overexpression(OE-MFN2).The cells were divided into control group,MFI8(20 μmol/L)group,VES groups(20 mg/L for MKN28 cells and 40 mg/L for MKN45 cells),VES+MFI8 group,OE-MFN2+MFI8 group and OE-MFN2+VES+MFI8 group.The MAMs were visualized by fluorescence microscopy,and the expression changes of MFN2,beclin-1 and LC3-II/I were detected by Western blot.RESULTS:The results of the CCK8 assay showed that VES significantly inhibited the viability of both human gastric can-cer cell lines(P<0.05).After VES treatment,the formation of typical autophagosomes and MAMs was observed in both cell lines by transmission electron microscopy.Fluorescence microscopy showed a significant increase in GFP signals of MAMs.Western blot analysis showed that with increasing doses of VES,the expression levels of MFN2,beclin-1,and LC3-II/I were significantly up-regulated,while that of p62 was significantly down-regulated(P<0.05).Compared with VES group,the cells pretreated with MFI8 followed by VES exposure showed markedly reduced GFP signals of MAMs and much lower protein levels of MFN2,beclin-1 and LC3-II/LC3-I(P<0.05).Transfection with an MFN2 overexpression plasmid rescued MFN2 expression.Compared with VES+MFI8 group,the cells in OE-MFN2+VES+MFI8 group had much higher protein expression levels of MFN2,beclin-1 and LC3-II/LC3-I(P<0.05).CONCLUSION:The VES may partici-pates in the regulation of autophagy in human gastric cancer cells by promoting the formation of MAMs.
2.Effect of vitamin E succinate on autophagy in human gastric cancer cells via mitochondria-associated endoplasmic reticulum membranes
Miaomiao CAO ; Fangyu CHEN ; Zhiyang WEI ; Mengmeng LÜ ; Ziqing XING ; Jinze WANG ; Shuang LI ; Liying HOU
Chinese Journal of Pathophysiology 2025;41(11):2157-2165
AIM:This study aims to investigate whether vitamin E succinate(VES)induces autophagy in hu-man gastric cancer cells through the promotion of mitochondria-associated endoplasmic reticulum membranes(MAMs).METHODS:Human gastric cancer cell lines MKN28 and MKN45 were cultured in vitro.Cell viability was assessed us-ing the CCK8 assay,and two cell growth curves were plotted to determine the treatment concentration of VES.Control groups,VES dose groups(MKN28:5,10,20,and 40 mg/L;MKN45:10,20,40,and 80 mg/L),an autophagy-posi-tive control group(rapamycin,RAPA,100 nmol/L),and a MAMs-positive control group(oligomycin A,10 mg/L)were set up.Cells were harvested after 24 h of treatment for subsequent experiments.The formation of autophagosomes and MAMs was observed using transmission electron microscopy.The expression levels of autophagy-related proteins,includ-ing beclin-1,LC3-II/LC3-I,and p62,were detected by Western blot.MAMs labeled with split green fluorescent protein(GFP)were visualized by fluorescence microscopy.The expression of mitofusin 2(MFN2),a key molecule of MAMs,was also detected by Western blot.To inhibit MFN2 specifically,the cells were treated with mitochondrial fusion inhibitor 8(MFI8)and simultaneously transfected with an MFN2 plasmid to achieve MFN2 overexpression(OE-MFN2).The cells were divided into control group,MFI8(20 μmol/L)group,VES groups(20 mg/L for MKN28 cells and 40 mg/L for MKN45 cells),VES+MFI8 group,OE-MFN2+MFI8 group and OE-MFN2+VES+MFI8 group.The MAMs were visualized by fluorescence microscopy,and the expression changes of MFN2,beclin-1 and LC3-II/I were detected by Western blot.RESULTS:The results of the CCK8 assay showed that VES significantly inhibited the viability of both human gastric can-cer cell lines(P<0.05).After VES treatment,the formation of typical autophagosomes and MAMs was observed in both cell lines by transmission electron microscopy.Fluorescence microscopy showed a significant increase in GFP signals of MAMs.Western blot analysis showed that with increasing doses of VES,the expression levels of MFN2,beclin-1,and LC3-II/I were significantly up-regulated,while that of p62 was significantly down-regulated(P<0.05).Compared with VES group,the cells pretreated with MFI8 followed by VES exposure showed markedly reduced GFP signals of MAMs and much lower protein levels of MFN2,beclin-1 and LC3-II/LC3-I(P<0.05).Transfection with an MFN2 overexpression plasmid rescued MFN2 expression.Compared with VES+MFI8 group,the cells in OE-MFN2+VES+MFI8 group had much higher protein expression levels of MFN2,beclin-1 and LC3-II/LC3-I(P<0.05).CONCLUSION:The VES may partici-pates in the regulation of autophagy in human gastric cancer cells by promoting the formation of MAMs.
3.Changes of collagen and MMP-1 in liver, lung and kidney during growth of mice.
Jianping GAO ; Yang ZHANG ; Fangyu XING ; Yingjun KONG ; Guifeng ZHANG
Chinese Journal of Biotechnology 2021;37(2):646-654
The high performance liquid chromatography (HPLC) and enzyme-linked immunoassay (ELISA) were used to investigate the changes of collagen and matrix metalloproteinase-1 (MMP-1) in liver, lung and kidney during growth process of mice. The mice from 0 to 18 weeks were used as the research objects. The contents and proportions of hydroxyproline (Hyp), which were used to calculate the collagen contents, in liver, lung and kidney of different weeks were analyzed with HPLC. The contents and activity of MMP-1 in liver, lung and kidney of different weeks were analyzed with ELISA. The results showed that the collagen contents in liver, lung, and kidney were different (Lung(COL)>Kidney(COL)>Liver(COL)), and they all increased first and then decreased with weeks. The collagen contents in liver, lung, and kidney reached the highest level in the ninth (5.52 ng/mg), sixth (54.10 ng/mg) and ninth (19.20 ng/mg) week, respectively. Then it declined slowly from 9 to 18 weeks. The result of ELISA showed that the MMP-1 contents in liver, lung and kidney decreased first and then increased with weeks, and the trend of MMP-1 activity was opposite. It indicated that the increase of collagen contents in the tissues will inhibit the secretion of MMP-1.
Animals
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Collagen
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Kidney
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Liver
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Lung
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Matrix Metalloproteinase 1
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Matrix Metalloproteinase 2
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Mice
4.User requirements of intelligent design for Intensive Care Delirium Screening Checklist
Xing REN ; Fangyu YANG ; Ying WU ; Yuling CHEN
Chinese Journal of Modern Nursing 2018;24(18):2147-2151
Objective?To determine and analyze the problems of nurses during using printed Intensive Care Delirium Screening Checklist (ICDSC) for evaluating through user requirements analysis, which is necessary for information systems development, so as to provide a basis for developing an ICDSC interactive strategy. Methods?Two methods, the documentation method and task check method, were used to analyze user requirements of printed ICDSC. The retrieval words of documentation method were "ICDSC" and "intensive care delirium screening checklist". This study retrieved documents on using and evaluation of ICDSC publishing in PubMed, Springer, Elsevier, CNIAHL, CNKI and WanFang Data from January 2000 to December 2017. The find what focused on description related to problems of Intensive Care Unit (ICU) nurses during using printed ICDSC. The task check method was used to select ICU nurses as user representatives to assess delirium of four patients cases provided by researchers using printed ICDSC. Each case all perfectly completed 8 appraising tasks from ICDSC assessments. Researchers observed and recorded problems of ICU nurses during fulfilling the task. Nurses' sample size was based on information saturation (till no new information occurred). Results?A total of 7 articles among 144 ones retrieved referred to problems of nurses during using printed ICDSC to assess delirium including time-consuming, failing to understand items, deficiency/inaccuracy in nursing record. A total of 15 ICU nurses were selected in task check method. Results showed that the problems included time-consuming, omitting items, failing to understand items, scoring mistakenly, deficiency/inaccuracy in nursing record. Conclusions?There are problems such as time-consuming, omitting items, failing to understand items, scoring mistakenly and assessed information without inaccurate source in using printed ICDSC. The interactive strategy should be developed according to these problems.
5.Application and prospect of big data in nursing field
Lili MA ; Fangyu YANG ; Xing REN ; Ting DENG
Chinese Journal of Modern Nursing 2017;23(3):308-312
With the widespread application of medical big data and the rapid development of nursing informatics, the application of big data in the service of nursing science induces high attention of nursing field. From the perspective of the concept of big data and their characteristics, this paper explained the technique of big data and its processing flow in details. Combined with five application fields of medical big data, this paper not only summarized the application situation of big data in nursing field, but discussed and analyzed the development prospect of nursing big data and its difficulty, which offers reference for the nursing academic circle.
6.Investigation of ICU delirium management status in two classⅢ grade A hospitals in Beijing
Jinglian LI ; Fangyu YANG ; Ying WU ; Ting DENG ; Xing REN
Chinese Journal of Modern Nursing 2017;23(18):2357-2361
Objective To investigate the current status of ICU delirium management so as to provide reference for the clinical management of ICU delirium.Methods A total of 6 ICU units from two class Ⅲ grade A hospitals in Beijing were selected by convenient sampling. The spot investigation method was used to investigate the delirium management status and effect from June to September 2016.Results Six ICU units had neither developed a complete delirium management system and standardized process, nor systematically evaluated delirium risk factors. When delirium patients had symptoms, delirium was diagnosed according to the clinical experience in 6 ICU units, while specialist consultation was used in 3 ICU units. None of these units used any delirium screening tool. The delirium prevention rate was from 46.2% to 76.9%. The implementation rate of delirium treatment was 65.8%. Among 38 cases of delirium patients diagnosed by medical staff, 25 cases (65.8%) of which were treated. During the study, there were 106 cases of delirium patients; the incidence of delirium was 30.8%; the delirium identification rate of medical staff was 35.8%.Conclusions The normalization of routine monitoring of delirium in ICU is necessary. Prevention and treatment for delirium is inadequate. The effect of delirium management needs to be further improved.

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