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Objective:To screen out the suitable nonpolar molecular cosolvent and concentration with adventitious root phenotype and ginsenoside content in the controlled experiment as the evaluation indexes， so as to lay a solid foundation for exploring the causes for good shape and high quality of <italic>Panax quinquefolium</italic>. Method:After being treated with different concentrations of dimethyl sulfoxide （DMSO） and ethanol， the adventitious roots were scanned using a panoramic scanner， and the resulting images were used for measuring the branch number and average diameter by WinRHIZO Pro 2016， Synbiosis ProtoCol 3 colony counter， Image J， and SmartRoot. The contents of ginsenosides Rg<sub>1</sub>， Rb<sub>1</sub>， and Re were determined by ultra-performance liquid chromatography tandem mass spectrometry （UPLC-MS/MS）. Result:Compared with the blank control， the 0.1% DMSO and 75% ethanol made the adventitious root phenotype and ginsenoside contents significantly changed. Specifically， the branch number and average diameter were significantly reduced. The ginsenoside Rg<sub>1</sub> in the adventitious roots decreased after 0.1% DMSO treatment， whereas the ginsenosides Rg<sub>1</sub> and Re increased after 75% ethanol treatment. The adventitious root phenotype and ginsenoside contents in the 0.1% DMSO treatment group were not significantly different from those in the control group. Conclusion:The 0.01% DMSO does not affect the adventitious root growth of <italic>P. quinquefolium </italic>and is insoluble in water， enabling it to be considered as a suitable nonpolar molecular cosolvent for future research on the genetic causes for the good shape and high quality of <italic>P. quinquefolium</italic>.
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Objective To explore the effect of different antifreeze or antioxidant at different cryopreservation conditions on the activity and the morphology of adipose cells.Methods After purification,fat granules were divided into 5 groups; the same volume of antifreeze or antioxidants were added in each group,stored at-20℃ (refrigerator),-80℃ (ultra-low-temperature refrigerator) and-196 ℃ (liquid nitrogen) ; the amount of glucose transfer was determined and the morphological changes were observed for each group after two weeks,two months,and three months.Results Fat activity of the group adding both dimethyl sulfoxide (DMSO) and trehalose was significantly higher than that in other groups at the same temperature and the same time point.The decreased amount of glucose transfer of 80℃ group and 196℃ group was significantly lower than that of 20℃ group,and that in groups of each 196℃ were greater than that in groups of each 80℃.Conclusions Antifreeze can obviously keep activity of fat cells.Preservative effect of DMSO is better than trehalose,and that of both combination is much better.Liquid nitrogen storage of adipose tissue for three months and fat granules stored at 80℃ for two months are still suitable for clinical use.
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Objective To search for the application of trehalose as cryoprotectant for keeping aortic valve homograft in liquid nitrogen.Methods The aortic valve homografts of Wistar rat were kept in 10%DMSO and divided into 2 groups(control and test group).The test group was added a supplement of trehalose 0.1mol?L~(-1) and all the hemografts were gradually hypothermied and then cryopreserved in-196℃ liquid nitrogen.After 6 months all the cryopreserved valves were thawed.The aortic valve homografts were observed by light and electron microscope.At the same time,the viability of endotheliocyte was measured.Results The viability of endotheliocyte in test group was higher than that in the control group and the structural damage was also much lighter.Conclusion The cryoprotective effect of combined use of 10%DMSO+0.1mol?L~(-1) trehalose was much better than that of 10%DMSO alone.
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Objective To observe the protective effects of various extracts from Flos Daturae (FD), including active fraction (AF-FD), withanolides constituents (WC-FD), and flavonoids constituents (FC-FD) on Chinese hamster ovary (CHO) cells against cytotoxicity induced by DMSO. Methods The survival rate of CHO cells was examined by MTT assay and LDH leakage assays. Results Using MTT assay, coincubation of CHO cells with 3% DMSO for 24 h resulted in a significant reduction of survival rate of CHO cells. AF-FD was tested in a range of 10—80 ?g/mL to improve the survival rate of CHO cells in a dose-dependent manner. FC-FD (2.5—20 ?g/mL), but not WC-FD (30—120 ?g/mL), could significantly relieve the injury induced by 3% DMSO in CHO cells. In the measurement of LDH leakage, coincubation of CHO cells with 4.5% DMSO for 24 h obviously increased LDH release. However, all the compounds tested, including AF-FD (10—80 ?g/mL), WC-FD (30—120 ?g/mL), and FC-FD (2.5—20 ?g/mL) had no effect on LDH leakage induced by 4.5% DMSO. Conclusion The findings suggest that the FC-FD may protect CHO cells from DMSO cytotoxicity assessed by MTT assay, which may be associated with improving mitochondrial function, but not protecting the membrane injury of CHO cells.