1."A preliminary establishment of the""single-cell-transplant-tumorigenic-test""model for identifying liver cancer stem cells"
Wei JIA ; Yanhui CEN ; Juan BAO ; Rui YANG ; Guozhen HE ; Xiaojun WU ; Yiyi LI
Chinese Journal of Tissue Engineering Research 2017;21(9):1324-1328
BACKGROUND: There is no effective method to identify liver cancer stem cells until now, which has become one of the major challenges in this field. OBJECTIVE: To explore a more effective and suitable way for the identification of liver cancer stem cells in hepatocellular carcinoma cell lines. METHODS: Firstly, the single cell separation technique was used to obtain single cells, which were seeded into 96-wel plates one by one. Secondly, cell sublines derived from single cell colonies (tumorigenic colonies) were selected and obtained. At last, the cells from these clones were transplanted into the forelimb armpits of nude mice to observe the tumorigenic ability. RESULTS AND CONCLUSION: The number of holes of single cells obtained was 371 by the single cell separation technique, and the success rate was 96.4%. According to the growth of cell clone, tumorigenic colonies were selected to be transplanted to the forelimb armpits of nude mice, and the tumor formation rate of the colonies was 100%. The identification model of single-cell-transplant-tumorigenic-test for liver cancer stem cells is confirmed to be preliminarily established, which lays the technology and methodological basis for the follow-up research.
2.Establishment and identification of pancreatic stem cell strain derived from islets of Kunming mice under feeder layer conditions
Yanhui CEN ; Rui YANG ; Wei JIA ; Zhonghua LI ; Zhenguo ZHONG ; Jing ZHONG ; Juan BAO ; Guozhen HE ; Xiaojun WU ; Yiyi LI
Chinese Journal of Tissue Engineering Research 2017;21(13):2087-2093
BACKGROUND:Until now, little has been reported on establishment of pancreatic stem cell strains and lines,and the purification of pancreatic stem cells is difficult since the cell line establish rate is low.OBJECTIVE:To explore a more rational and effective technique of in vitro separation and continuous passage of pancreatic stem cells, with the hope to establish cell strains and even cell lines and to lay the foundation for the follow-up study of pancreatic stem cells in the treatment of diabetes.METHODS:Firstly, Percoll discontinuous density gradient centrifugation method was applied to separate the mouse pancreatic endocrine portion from the exocrine portion, then to obtain cell strains with highly proliferative ability and low differentiation from pancreatic endocrine portion-the islet. We used mouse embryonic fibroblasts treated with mitomycin C as a feeder layer, for in vitro continuous culture of islet-derived pancreatic stem cells under feeder layer conditions until they were transferred to the 30th passage to establish cell lines. Then pancreatic stem cell line derived from pancreatic islet was detected and identified by a series of tests including growth characteristic test, morphological observation, related molecular marker identification and differentiation characteristic identification.RESULTS AND CONCLUSION:In the continuous process of passage, pancreatic stem cells showed active proliferative ability, and maintained the typical morphological characteristics of stem cells and expression of pancreatic stem cell marker-Nestin. After induction, pancreatic stem cells showed insulin gene expression,reflecting their differentiation potential. Therefore, under the condition of feeder layer, the pancreatic stem cell line derived from Kunming mice was successfully established and the related identification was completed,which lays the foundation for the following research.
3.Long-term culture and differentiation of neural stem cells of embryonic mice.
Liye YANG ; Guozhen HUI ; Dashi BAO ; Lizhen JIANG ; Jian FEI ; Lihe GUO
Chinese Journal of Surgery 2002;40(10):783-785
OBJECTIVESTo assess the culture and differentiation of neural stem cells in embryonic mice and set up a basis for further research in to neural stem cells.
METHODSEmbryonic cortices of mice were dissociated and single cell suspensions were achieved by mechanical methods in sterile conditions, and cells were seeded in uncoated plate in N2 medium. The cells were passaged by mechanical methods, frozen and thawed by general procedure. They were identified by immunocytochemical techniques.
RESULTSNeural stem cells from embryonic mice were successfully cultured forming typical neurospheres in suspension. Neurons, astrocytes and oligodendrocytes were differentiated from neural stem cells, with a ratio of 7%, 85% - 90% and 2% - 4% respectively.
CONCLUSIONSNeural stem cells, which can be cultured and passaged steadily in vitro and they are the ideal cell sources for cell transplantation and gene therapy.
Animals ; Cell Differentiation ; Cells, Cultured ; Embryo, Mammalian ; cytology ; Immunohistochemistry ; Mice ; Mice, Inbred BALB C ; Neurons ; cytology ; Stem Cells ; cytology
4.Summary of best evidence for exercise rehabilitation for atrial fibrillation
Jie WANG ; Guozhen SUN ; Min GAO ; Zhipeng BAO ; Lin WANG
Chinese Journal of Practical Nursing 2022;38(19):1465-1471
Objective:To evaluate and summarize the best evidence of exercise instructions for patients with atrial fibrillation.Methods:A comprehensive search about evidence on exercise rehabilitation in patients with atrial fibrillation was conducted in following databases: Joanna Briggs Institute (JBI) Library, The Agency for Healthcare Research and Quality (AHRQ), National Institute for Health and Clinic Excellence (NICE), Scottish Intercollegiate Guidelines Network (SIGN), Registered Nurses′ Association of Ontario (RNAO), Physiotherapy Evidence Database (PEDro), American College of Physicians (ACP), Cochrane Library, PubMed, Embase, CINAHL, Medlive, China National Knowledge Infrastructure (CNKI), WanFang Database, VIP Database, and China Biology Medicine (CBM). The retrieval period was from the inception of databases to January 2021. Two researchers with evidence-based nursing background assessed the quality of the literature independently and identified the level of evidence.Results:A total of 16 articles were included, including 4 guidelines, 4 systematic reviews, 4 expert consensuses, 4 randomized controlled trials. Combined with professional judgment, 35 pieces of best evidence in 7 aspects were summarized, including exercise benefits, the appropriate crowd, exercise evaluation, exercise mode, exercise intensity, exercise time, exercise supervision and safety.Conclusions:Regular exercise is safe and beneficial for patients with atrial fibrillation. Clinical staff should guide patients to exercise moderately by applying the best evidence with scientific exercise intervention, promoting patients′ physical and mental health.
5.Path analysis of the selection and evaluation of abroad-study based on the effectiveness of scientific research
Danlu ZHANG ; Guozhen BAO ; Qi WANG
Chinese Journal of Medical Science Research Management 2020;33(3):161-165
Objective:To explore the combination of study abroad and scientific research effectiveness, improve the " effectiveness concern" related to the selection mechanism for abroad-study, establish an evaluation system combined studying abroad and scientific research effectiveness.Methods:The research was based on 45 local-funded abroad-study programs, and used path analysis to connect main influence factors of studying abroad with scientific achievements, and establish a model.Results:The final path analysis model contained two regression equations: (1) in the equation of SCI articles, adjusted R2=0.600, F=5.698, P<0.001, statistic difference was significant. In this pathway, influence factors on SCI articles were successively Professional Title ( β=0.736, P<0.001), Age ( β=-0.700, P<0.001) and foreign institutions ( β=-0.318, P=0.031); (2) in the equation of Project funding, adjusted R2=0.490, F=7.608, P<0.001, statistic difference was significant. In this pathway, influence factors on project funding were successively SCI Articles ( β=0.409, P=0.010), Study country ( β=-0.342, P=0.015), and Degree ( β=-0.223, P=0.043). Conclusions:According to the path analysis, impacts of studying abroad on scientific research effectiveness were evaluated and detailed more precisely. The research established scientific-effectiveness oriented pathway between assigning and evaluation system, providing support to promote effectiveness of hospital abroad-study scientific outputs, and provide possible reference for local and national governments to optimize international exchange programs.