Specific oligonucleotides such as telomere DNA and aptamer often undergo conformational changes upon ligand binding. Composite reagent composed of o-phthalaldehyde and β-mercaptoethanol（OPAME） has been extensively ...Specific oligonucleotides such as telomere DNA and aptamer often undergo conformational changes upon ligand binding. Composite reagent composed of o-phthalaldehyde and β-mercaptoethanol（OPAME） has been extensively applied to fluorescent detection of amino compounds based on the reaction of primary amino-group, herein we proposed a general spectrofluorometry for ions and small molecules due to conformational changes upon ligand binding taking K^＋ and ATP as examples. In a borate controlled buffer medium, telomere DNA could react with OPAME, giving a thio-subtituted isoindole compound with strong fluorescence emission at 455 nm when excited at 340 nm. It was found that however, the fluorescence emission was greatly reduced in the presence of K^＋ since the formation of the quadruplex structure inhibits the reaction activity of amino-groups of telomere DNA. In order to testify the general application of OPAME reagent based on the conformational change of oligonucleotides, we further proposed a sensitive method of ATP based on its highly selective interaction with ATP-aptamer. The above mentioned applications show that the spectrofluorometry with the aid of OPAME reagent is simple, label free that is expected to be potentially general for DNA conformational change-based target detection.展开更多
β-mercaptoethanol can induce adipose-derived stromal cells to rapidly and efficiently differentiate into neurons in vitro.However,because of the short survival time of the differentiated cells,clinical applications f...β-mercaptoethanol can induce adipose-derived stromal cells to rapidly and efficiently differentiate into neurons in vitro.However,because of the short survival time of the differentiated cells,clinical applications for this technique are limited.As such,we examined apoptosis of neurons differentiated from adipose-derived stromal cells induced with β-mercaptoethanol in vitro using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling and transmission electron microscopy.The results revealed that the number of surviving cells decreased and apoptosis rate increased as induction time extended.Taken together,these results suggest that apoptosis occurring in the process of adipose-derived stromal cells differentiating into neurons is the main cause of cell death.However,the mechanism underlying cellular apoptosis should be researched further to develop methods of controlling apoptosis for clinical applications.展开更多
This study was conducted to investigate the effects of different concentrations ofβ-mercaptoethanol on the development ability of oocytes in Dorper sheep at the age of 6 weeks.In this study,6 Dorper sheep of 6 weeks ...This study was conducted to investigate the effects of different concentrations ofβ-mercaptoethanol on the development ability of oocytes in Dorper sheep at the age of 6 weeks.In this study,6 Dorper sheep of 6 weeks old were chosen to induce oocyte collection in vivo by gonadotropin,and A and B grade cumulus oocyte complexes were obtained(COCs)with different concentrations of(0,50,70 and 100μmol/L)ofβ-mercaptoethanol in the mature liquid.The cleavage rate and blastocyst rate were counted after matured oocytes and capacitated sperm were co-incubated and fertilized eggs were cultivated.The results showed that compared with the cleavage rate(59.13%)and the blastocyst rate(12.17%)of the control group,the cleavage rates(60.87%,63.48%and 65.22%)and blastocyst rates(14.78%,17.39%and 21.74%)of the test groups I,II and III were higher(P>0.05),and the order was test group III>test group II>test group I>control group.The cleavage rate and blastocyst rate of the 100μmol/Lβ-mercaptoethanol group increased by 6.09 percentage points and 9.57 percentage points,respectively.It indicated that the addition of a certain amount ofβ-mercaptoethanol in the mature liquid could improve the developmental ability of oocytes and the quality of fertilized eggs of the 6-week-old Doper sheep.When theβ-mercaptoethanol concentration was within 100μmol/L,the cleavage rate and the blastocyst rate of oocytes were on the rise.The optimal concentration ofβ-mercaptoethanol in the mature liquid still needs further study.展开更多
[Objective]The aim was to explore optimization of system of oocytes in-vitro culture of young animals. [Method]Effects of EGF （ epidermal growth factor） and β-mercaptoethanol in maturation media on fertilization,cl...[Objective]The aim was to explore optimization of system of oocytes in-vitro culture of young animals. [Method]Effects of EGF （ epidermal growth factor） and β-mercaptoethanol in maturation media on fertilization,cleavage and blastaea were researched,which were then compared with those of adult sheep. [Result]EGF in different concentrations had little effects on rates of cleavage and blastaea （ P 〉0.05） and β-mer-captoethanol in different concentrations would improve blastaea rate of oocytes,for example,100 μmol/L of β-mercaptoethanol has significant effects on balstaea rate （ P 〈0.05） ,but has little effects on cleavage rate （ P 〉0. 05） . In addition,rates of cleavage and balstaea of oocytes in lamb were both lower than those of adult sheep （ P 〈0.05） ; fertilization rate of oocytes in lamb （ P 〈0.05） ,which differed little with that of adult sheep （ P 〉0.05） ,could be significantly enhanced by 100 μmol/L of β-mercaptoethanol. Furthermore,polyspermy rate was higher than that of adult sheep without β-mercaptoethanol （ P 〈0.05） ; the rate was of little differences with that of adult sheep with 100 μmol/L of β-mercaptoethanol （ P 〉 0.05） ; unfertilization rate （ 20%） in media without β-mercaptoethanol was a little higher （ P 〉0.05） than those of adult sheep （ 12.3%） and those in media with β-mercaptoethanol （ 13.5%） . [Conclusion]Developmental capacity of oocytes and fertilization rate could be improved by 100 μmol/L of β-mercaptoethanol with polyspermy rate reduced,but developmental capacity of lamb was significantly lower than that of adult sheep.展开更多
基金Supported by the National Natural Science Foundation of China(No.20775061)
文摘Specific oligonucleotides such as telomere DNA and aptamer often undergo conformational changes upon ligand binding. Composite reagent composed of o-phthalaldehyde and β-mercaptoethanol（OPAME） has been extensively applied to fluorescent detection of amino compounds based on the reaction of primary amino-group, herein we proposed a general spectrofluorometry for ions and small molecules due to conformational changes upon ligand binding taking K^＋ and ATP as examples. In a borate controlled buffer medium, telomere DNA could react with OPAME, giving a thio-subtituted isoindole compound with strong fluorescence emission at 455 nm when excited at 340 nm. It was found that however, the fluorescence emission was greatly reduced in the presence of K^＋ since the formation of the quadruplex structure inhibits the reaction activity of amino-groups of telomere DNA. In order to testify the general application of OPAME reagent based on the conformational change of oligonucleotides, we further proposed a sensitive method of ATP based on its highly selective interaction with ATP-aptamer. The above mentioned applications show that the spectrofluorometry with the aid of OPAME reagent is simple, label free that is expected to be potentially general for DNA conformational change-based target detection.
文摘β-mercaptoethanol can induce adipose-derived stromal cells to rapidly and efficiently differentiate into neurons in vitro.However,because of the short survival time of the differentiated cells,clinical applications for this technique are limited.As such,we examined apoptosis of neurons differentiated from adipose-derived stromal cells induced with β-mercaptoethanol in vitro using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling and transmission electron microscopy.The results revealed that the number of surviving cells decreased and apoptosis rate increased as induction time extended.Taken together,these results suggest that apoptosis occurring in the process of adipose-derived stromal cells differentiating into neurons is the main cause of cell death.However,the mechanism underlying cellular apoptosis should be researched further to develop methods of controlling apoptosis for clinical applications.
基金Supported by Earmarked Fund for Construction of National Wool Sheep Industry Technology Research System(CARS-39-24)Fund for Science and Technology Innovation Team in Shanxi Province(201705D131028-20)
文摘This study was conducted to investigate the effects of different concentrations ofβ-mercaptoethanol on the development ability of oocytes in Dorper sheep at the age of 6 weeks.In this study,6 Dorper sheep of 6 weeks old were chosen to induce oocyte collection in vivo by gonadotropin,and A and B grade cumulus oocyte complexes were obtained(COCs)with different concentrations of(0,50,70 and 100μmol/L)ofβ-mercaptoethanol in the mature liquid.The cleavage rate and blastocyst rate were counted after matured oocytes and capacitated sperm were co-incubated and fertilized eggs were cultivated.The results showed that compared with the cleavage rate(59.13%)and the blastocyst rate(12.17%)of the control group,the cleavage rates(60.87%,63.48%and 65.22%)and blastocyst rates(14.78%,17.39%and 21.74%)of the test groups I,II and III were higher(P>0.05),and the order was test group III>test group II>test group I>control group.The cleavage rate and blastocyst rate of the 100μmol/Lβ-mercaptoethanol group increased by 6.09 percentage points and 9.57 percentage points,respectively.It indicated that the addition of a certain amount ofβ-mercaptoethanol in the mature liquid could improve the developmental ability of oocytes and the quality of fertilized eggs of the 6-week-old Doper sheep.When theβ-mercaptoethanol concentration was within 100μmol/L,the cleavage rate and the blastocyst rate of oocytes were on the rise.The optimal concentration ofβ-mercaptoethanol in the mature liquid still needs further study.
基金funded by the National Natural Science Foundation of China (31160460)Doctor Fund of Xinjiang Military Groups (2010JC10)+3 种基金National Wool Industry System (nycytx-40-06)National Key Technology Research and Development Program of the Ministry of Science and Technology of China(2011BAD28B05-1-1)863 Plan (2011AA100307)Breeding Strategy of Xinjiang Group (2011BA006)
文摘[Objective]The aim was to explore optimization of system of oocytes in-vitro culture of young animals. [Method]Effects of EGF （ epidermal growth factor） and β-mercaptoethanol in maturation media on fertilization,cleavage and blastaea were researched,which were then compared with those of adult sheep. [Result]EGF in different concentrations had little effects on rates of cleavage and blastaea （ P 〉0.05） and β-mer-captoethanol in different concentrations would improve blastaea rate of oocytes,for example,100 μmol/L of β-mercaptoethanol has significant effects on balstaea rate （ P 〈0.05） ,but has little effects on cleavage rate （ P 〉0. 05） . In addition,rates of cleavage and balstaea of oocytes in lamb were both lower than those of adult sheep （ P 〈0.05） ; fertilization rate of oocytes in lamb （ P 〈0.05） ,which differed little with that of adult sheep （ P 〉0.05） ,could be significantly enhanced by 100 μmol/L of β-mercaptoethanol. Furthermore,polyspermy rate was higher than that of adult sheep without β-mercaptoethanol （ P 〈0.05） ; the rate was of little differences with that of adult sheep with 100 μmol/L of β-mercaptoethanol （ P 〉 0.05） ; unfertilization rate （ 20%） in media without β-mercaptoethanol was a little higher （ P 〉0.05） than those of adult sheep （ 12.3%） and those in media with β-mercaptoethanol （ 13.5%） . [Conclusion]Developmental capacity of oocytes and fertilization rate could be improved by 100 μmol/L of β-mercaptoethanol with polyspermy rate reduced,but developmental capacity of lamb was significantly lower than that of adult sheep.
