ABSTRACT
BACKGROUND: Y-27632 is a Rho-associated coil-formed protein kinase inhibitor that can regulate the self-renewal of stem cells, promote clonal formation and cell survival, and regulate and protect neuronal cell growth and development. How to improve the differentiation efficiency of embryonic stem cells into neuron-like cells is highly important for nerve injury repair and nerve regeneration. OBJECTIVE: To investigate the effect of Y-27632 on the differentiation efficiency and function of human embryonic stem cells into neuron-like cells. METHODS: After resuscitation, human embryonic stem cells at passage 16 were subjected to morphological observation and staining identification. The embryoid bodies were prepared by suspension culture, and after 8 days of incubation, the cells were cultured in Sato medium containing different concentrations of Y-27632 (0, 5, 10, 20, 40 μmol/L) for 10 days and identification by staining. After induction for 18 days, the differentiation efficiency and neurite outgrowth were identified by immunofluorescence staining. RESULTS AND CONCLUSION: The human embryonic stem cells co-expressed Oct4 and SSEA-3 stem cell specific markers. After suspension culture for 8 days and further adherent culture for 10 days, the cells could be differentiated into neuron-like cells with neurogenic morphology and expressing Tuj-1. Y-27632, especially at a concentration of 10 μmol/L, not only promoted cell proliferation (a significant increase in adherent cells an Tuj-1 positive cells), but also facilitated cell differentiation into neurons. Immunofluorescence staining findings showed that 10 μmol/L Y-27632 significantly increased the number of Tuj-1 positive cells and neurites and the length of neurites after 18 days of differentiation. These results indicate that Y-27632 not only promotes the differentiation of human embryonic stem cells into neuron-like cells, but also accelerates neurite outgrowth.
ABSTRACT
Objective To investigate question comprehension and production among agrammatic aphasics, and to explore the mechanisms of any dysfunction in questioning.Methods Twenty aphasics were recruited in this study.According to the Chinese Agrammatism Battery,10 were classified as agrammatic (the agrammatic group) and 10 as non-agrammatic (the non-agrammatic group).Ten normal subjects served as a control group.All the subjects were tested in terms of their comprehension and production of questions using a set of general and what-where-who- why questions (wh-questions).Results No significant difference was found between the two experimental groups with regard to the correct comprehension and production of both general and wh-questions.However,there was a sig- nificant difference in correctness between comprehension and production.The performance of the agrammatic aphasics was worse than that of the non-agramatics and the normal subjects.Conclusion The impaired question comprehen- sion and production of Chinese agrammatic aphasics has its own characteristies which can form a basis for rehabilita- tion planning and outcome prediction.