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
In order to further understand the effects of drought stress on hydraulic characteristics and the relationship between hydraulic conductivity and aquaporins or water channels of root systems of three wolfberry cultiva...In order to further understand the effects of drought stress on hydraulic characteristics and the relationship between hydraulic conductivity and aquaporins or water channels of root systems of three wolfberry cultivars(Lycium chinense Mill.),hydraulic conductivity of 2-yearold pot-grown seedlings was measured under drought stress,rewatering,and treatment with exogenous mercuric chloride andβ-mercaptoethanol.Under moderate and severe drought stress levels,the most significant decrease of hydraulic conductivity was 37.3%and 24.0%,respectively,in the‘Ningqi 5’cultivar compared with the nonstressed control.After rewatering,the rate of recovery in specific conductivity was most rapid in the‘Mengqi 1’cultivar,at 0.058 and 0.072 kg MPa^-1 m^-2 s^-1 h^-1 under moderate and severe drought stress levels,respectively.The‘Mengqi 1’cultivar had the highest recovery degree of hydraulic conductivity under two concentrations ofβ-ME(500 or 1000 lmol L^-1),reaching 82.4%and 88.5%,respectively,of the initial conductivity.The adaptive capacity of hydraulic conductivity in the‘Ningqi 5’cultivar was weaker than in the‘Ningqi 1’and‘Mengqi 1’cultivars under drought stress.The recovery capacity of hydraulic conductivity in‘Mengqi 1’cultivar was stronger than the‘Ningqi 1’and‘Ningqi 5’cultivars after rewatering.Aquaporins of the‘Ningqi 1’cultivar root systems had the highest binding affinity with mercuric chloride,which was the most likely cause in the decrease in hydraulic conductivity,whereas aquaporins of‘Mengqi 1’root systems had the weakest binding affinity.The inhibitory effect of mercuric chloride was readily eliminated byβ-mercaptoethanol in the‘Mengqi 1’cultivar.The hydraulic characteristics of this cultivar were more sensitive to drought,mercuric chloride andβ-mercaptoethanol than the other cultivars.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
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
Objective To study the protective effects of hydroxysafflor yellow A (HSYA) against the oxidative damage caused by β-mercaptoethanol (BME) during neural differentiation of mesenchymal stem cells (MSCs) in vitro...Objective To study the protective effects of hydroxysafflor yellow A (HSYA) against the oxidative damage caused by β-mercaptoethanol (BME) during neural differentiation of mesenchymal stem cells (MSCs) in vitro. Methods When the confluence reached 50%-60%, 4th passage MSCs were divided into three groups to culture. Gt : normal group which was cultured using basic medium (DMEM containing 10% FBS all the time); G2: unprotected group which was continuously cultured using basic medium for 24 h, and then cultured using pre-induction medium (DMEM containing 10% FBS and 1 mmol/L BME); G3: protected group which was firstly cultured using protective medium (DMEM containing 10% FBS and 160 mg/L HSYA) for 24 h, and then cultured using pre-induction medium for 24 h. After these treatments as above, cell viability, relative levels of SOD/GSH and apoptosis rate were respectively detected. The expression of Bcl and Bax was examined by Western blotting. After HSYA protection and BME pre-induction, neural induction was performed. The expression of NSE and MAP-2 was respectively analyzed on cellular and molecular levels. Results Compared with unprotected group, 160 mg/L HSYA could obviously improve cells viability, maintain high level of SOD and GSH in MSCs, reduce apoptosis rate and improve the ratio of Bcl/Bax. After protection with 160 mg/L HSYA, the survival time of neuron-like cells could be extended. Immunocytochemical staining showed that after 10 h of neural induction, the differentiated neuron-like cells in protected group were still in a good state, and the mRNA levels of NSE and MAP-2 were increased during the induction course checked. Conclusion HSYA could improve the resistance of cells to the oxidative damage caused by BME.展开更多
β-mercaptoethanol induces in vitro adult adipose-derived stromal cells (ADSCs) to differentiate into neurons. However, the ultrastructural features of the differentiated neuronal-like cells remain unknown. In the p...β-mercaptoethanol induces in vitro adult adipose-derived stromal cells (ADSCs) to differentiate into neurons. However, the ultrastructural features of the differentiated neuronal-like cells remain unknown. In the present study, inverted phase contrast microscopy was utilized to observe β-mercaptoethanol-induced differentiation of neuronal-like cells from human ADSCs, and immunocytochemistry and real-time polymerase chain reaction were employed to detect expression of a neural stem cells marker (nestin), a neuronal marker (neuron-specific enolase), and a glial marker (glial fibrillary acidic protein). In addition, ultrastructure of neuronal-like cells was observed by transmission election microscopy. Results revealed highest expression rate of nestin and neuron-specific enolase at 3 and 5 hours following induced differentiation; cells in the 5-hour induction group exhibited a neuronal-specific structure, i.e., Nissl bodies. However, when induction solution was replaced by complete culture medium after 8-hour induction, the differentiated cells reverted to the fibroblast-like morphology from day 1. These results demonstrate that β-mercaptoethanol-induced ADSCs induced differentiation into neural stem cells, followed by morphology of neuronal-like cells. However, this differentiation state was not stable.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.
基金the National Natural Science Foundation of China(Grant No.31560380)Chief Expert of Modern Agricultural Traditional Chinese Medicine Industry System in Gansu Province(GARS-ZYC-1)the Open Project of Qinghai Key Laboratory of Qinghai-Tibet Plateau Biological Resources(Grant No.2017-ZJ-Y10)。
文摘In order to further understand the effects of drought stress on hydraulic characteristics and the relationship between hydraulic conductivity and aquaporins or water channels of root systems of three wolfberry cultivars(Lycium chinense Mill.),hydraulic conductivity of 2-yearold pot-grown seedlings was measured under drought stress,rewatering,and treatment with exogenous mercuric chloride andβ-mercaptoethanol.Under moderate and severe drought stress levels,the most significant decrease of hydraulic conductivity was 37.3%and 24.0%,respectively,in the‘Ningqi 5’cultivar compared with the nonstressed control.After rewatering,the rate of recovery in specific conductivity was most rapid in the‘Mengqi 1’cultivar,at 0.058 and 0.072 kg MPa^-1 m^-2 s^-1 h^-1 under moderate and severe drought stress levels,respectively.The‘Mengqi 1’cultivar had the highest recovery degree of hydraulic conductivity under two concentrations ofβ-ME(500 or 1000 lmol L^-1),reaching 82.4%and 88.5%,respectively,of the initial conductivity.The adaptive capacity of hydraulic conductivity in the‘Ningqi 5’cultivar was weaker than in the‘Ningqi 1’and‘Mengqi 1’cultivars under drought stress.The recovery capacity of hydraulic conductivity in‘Mengqi 1’cultivar was stronger than the‘Ningqi 1’and‘Ningqi 5’cultivars after rewatering.Aquaporins of the‘Ningqi 1’cultivar root systems had the highest binding affinity with mercuric chloride,which was the most likely cause in the decrease in hydraulic conductivity,whereas aquaporins of‘Mengqi 1’root systems had the weakest binding affinity.The inhibitory effect of mercuric chloride was readily eliminated byβ-mercaptoethanol in the‘Mengqi 1’cultivar.The hydraulic characteristics of this cultivar were more sensitive to drought,mercuric chloride andβ-mercaptoethanol than the other cultivars.
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.
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Major Projection of Hebei North University(No.ZD201413)
文摘Objective To study the protective effects of hydroxysafflor yellow A (HSYA) against the oxidative damage caused by β-mercaptoethanol (BME) during neural differentiation of mesenchymal stem cells (MSCs) in vitro. Methods When the confluence reached 50%-60%, 4th passage MSCs were divided into three groups to culture. Gt : normal group which was cultured using basic medium (DMEM containing 10% FBS all the time); G2: unprotected group which was continuously cultured using basic medium for 24 h, and then cultured using pre-induction medium (DMEM containing 10% FBS and 1 mmol/L BME); G3: protected group which was firstly cultured using protective medium (DMEM containing 10% FBS and 160 mg/L HSYA) for 24 h, and then cultured using pre-induction medium for 24 h. After these treatments as above, cell viability, relative levels of SOD/GSH and apoptosis rate were respectively detected. The expression of Bcl and Bax was examined by Western blotting. After HSYA protection and BME pre-induction, neural induction was performed. The expression of NSE and MAP-2 was respectively analyzed on cellular and molecular levels. Results Compared with unprotected group, 160 mg/L HSYA could obviously improve cells viability, maintain high level of SOD and GSH in MSCs, reduce apoptosis rate and improve the ratio of Bcl/Bax. After protection with 160 mg/L HSYA, the survival time of neuron-like cells could be extended. Immunocytochemical staining showed that after 10 h of neural induction, the differentiated neuron-like cells in protected group were still in a good state, and the mRNA levels of NSE and MAP-2 were increased during the induction course checked. Conclusion HSYA could improve the resistance of cells to the oxidative damage caused by BME.
文摘β-mercaptoethanol induces in vitro adult adipose-derived stromal cells (ADSCs) to differentiate into neurons. However, the ultrastructural features of the differentiated neuronal-like cells remain unknown. In the present study, inverted phase contrast microscopy was utilized to observe β-mercaptoethanol-induced differentiation of neuronal-like cells from human ADSCs, and immunocytochemistry and real-time polymerase chain reaction were employed to detect expression of a neural stem cells marker (nestin), a neuronal marker (neuron-specific enolase), and a glial marker (glial fibrillary acidic protein). In addition, ultrastructure of neuronal-like cells was observed by transmission election microscopy. Results revealed highest expression rate of nestin and neuron-specific enolase at 3 and 5 hours following induced differentiation; cells in the 5-hour induction group exhibited a neuronal-specific structure, i.e., Nissl bodies. However, when induction solution was replaced by complete culture medium after 8-hour induction, the differentiated cells reverted to the fibroblast-like morphology from day 1. These results demonstrate that β-mercaptoethanol-induced ADSCs induced differentiation into neural stem cells, followed by morphology of neuronal-like cells. However, this differentiation state was not stable.