[Objective] A method for the determination of exchangeable Ca and Mg contents in tobacco growing area by atomic absorption spectrometry was established. [Method] Factors in the extraction process, sample weight, ammon...[Objective] A method for the determination of exchangeable Ca and Mg contents in tobacco growing area by atomic absorption spectrometry was established. [Method] Factors in the extraction process, sample weight, ammonium acetate volume, extraction time and strontium chloride volume were tested, and the stability, precision and accuracy of the method were also investigated. Meanwhile, samples at different pH values were determined by this method. [Result] The results showed that this method operation has simple operation with good stability and high precision. [Conclusion] This method could be applied to determine exchangeable Ca and Mg contents in tobacco growing area.展开更多
A leaching experiment was carried out with repacked soil columns in laboratory to stndy the leachingprocess of a red soil derived from sandstone as affected by warions fertilization practices. The treatments wereCK (a...A leaching experiment was carried out with repacked soil columns in laboratory to stndy the leachingprocess of a red soil derived from sandstone as affected by warions fertilization practices. The treatments wereCK (as a control), CaCO_3, CaSO_4, MgCO_3, Ca(H_2PO_4)_2, Urea, KCl, Mnltiple (a mixture of the above-mentioned fertilizers), and KNO_3. The fertilizers were added to the bare surface of the soil columns, andthen the columns were leached with 120 mL deionized water daily through peristaltic pumps over a periodof 92 days. At the end of leaching process, soils were sampled from different depths of the soil profiles, i.e.,0-5 cm, 5-10 cm, 10-20 cm, 20-40 cm, and 40-60 cm. The results showed when applying Ca, Mg, andK to the bare surface of the soil columns, exchangeable Ca￣(2+), Mg￣(2+), and K￣+ in the upper layer of thesoil profile increased correspondingly, with an extent depending mainly on the application rates of Ca, Mg,and K and showing a downward trend. CaCO_3, CaSO_4, MgCO_3, and Ca(H_2PO_4)_2 treatments had scarcelyany effect on movement of exchangeable K￣+, while CaCO_3, and CaSO_4 treatments significantly promotedthe downward movement of exchangeable Mg￣(2+) although these two treatments had no obvious effect onleaching losses of Mg. The fact that under Urea treatment, exchangeable Ca￣(2+) and Mg￣(2+) were higheras compared to CK treatment showed urea could prevent leaching of exchangeable Ca￣(2+) and Mg￣(2+). Theobvious downward movement of exchangeable Ca￣(2+) and Mg￣(2+) was noticed in KCl treatment. In Multipletreatment, the downward movement of exchangeable Ca￣(2+) and Mg￣(2+) was evident, while that of K￣+ was lessevident. Application of KNO_3 strongly promoted the downward movement of exchangeable Ca￣(2+) and Mg￣(2+)in the soil profile.展开更多
To determine the effects of HOE 694, a new and potent Na＋- H＋ exchanger blocker, on transient inward current (Iti) and Na＋- Ca2＋ exchange during hypoxia- reoxygenation in guinea pig cardiomyocytes. Methods. Cardio...To determine the effects of HOE 694, a new and potent Na＋- H＋ exchanger blocker, on transient inward current (Iti) and Na＋- Ca2＋ exchange during hypoxia- reoxygenation in guinea pig cardiomyocytes. Methods. Cardiomyocytes were isolated from adult guinea pig ventricle. Experiment was performed in an experimental chamber that allowed the cells to be exposed to a sufficiently low O2 pressure. The cells were subjected to hypoxia and reoxygenation. The ionic currents were studied with patch clamp technique. Results. In the absence of HOE 694, hypoxia- reoxygenation induced Iti in 12 of 15 experiments; but in cardiomyocytes pretreated with HOE 694 (10～ 50μ mol/L), the incidence of Iti observed during reoxygenation was reduced to 5 of 11 experiments and 3 of 10 experiments, P Conclusions. Blockade of the Na＋- H＋ exchange by HOE 694 could reduce Ca2＋ overload upon hypoxia- reoxygenation, and inhibition of Na＋- H＋ exchange may also indirectly decrease Na＋- Ca2＋ exchange activity during hypoxia.展开更多
We determined whether La3+ enter human peripheral blood lymphocytes via Na+/Ca2+ exchanger (measured with fura-2). We first compared the sensitivity of fura-2 with La3+ and Ca2+, the result indicates that the sensitiv...We determined whether La3+ enter human peripheral blood lymphocytes via Na+/Ca2+ exchanger (measured with fura-2). We first compared the sensitivity of fura-2 with La3+ and Ca2+, the result indicates that the sensitivity of fura-2 for La3+ is much greater than for Ca2+. La3+ forms a 1:1 La3+-fura-2 complex (apparent dissociation constant = 1.7x10(-12) mol/L, pH 7.05). Ouabain-pretreated cells, suspended in Na+-free medium, showed that La3+ can enter human lymphocytes via the Na-i(+)/Ca2+ (La3+)(o) exchanger and is found to be about 10(-12) mol/L in cells exposed to 0.4 mmol/L La3+. Otherwise, the higher concentration (0.1 mmol/L) blocks the Na-i(+)/Ca2+(La3+)(o) exchange-mediated influx of Ca2+, but the lower concentration (0.01 mmol/L) appears to increase Ca2+ entry.展开更多
At present, there are relevant scientific materials on the cellular and molecular mechanisms of electrogenic Na/K pump function and structure, as well as on the potential- and ligand-activated ionic channels in the me...At present, there are relevant scientific materials on the cellular and molecular mechanisms of electrogenic Na/K pump function and structure, as well as on the potential- and ligand-activated ionic channels in the membrane. However, the role of electrogenic Na/K pump in regulation of semipermeable properties of cell membrane has not been elucidated yet, which is due to the fact that our knowledge about the biophysical properties of cell membrane is based on the conductive membrane theory of Hodgkin-Huxley-Katz, which is developed on internally perfused squid axon and lacks intracellular metabolism. Thus, the accumulated abundance of data on the role of G-proteins-dependent intracellular signaling system in regulation of Na/K pump activity and biophysical properties of cell membrane presumes fundamental revision of some statements of membrane theory. The aim of the present review is to briefly demonstrate our and literature data on cell hydration-induced auto-regulation of Na/K pump as well as on its role in metabolic control of semipermeable properties and excitability of neuronal membrane, which are omitted in the study of internally perfused squid axon.展开更多
CRISPR-Cas9 system is now widely used to edit a target genome in animals and plants. Cas9 protein derived from Streptococcus pyogenes（Sp Cas9） cleaves double-stranded DNA targeted by a chimeric single-guide RNA（sg ...CRISPR-Cas9 system is now widely used to edit a target genome in animals and plants. Cas9 protein derived from Streptococcus pyogenes（Sp Cas9） cleaves double-stranded DNA targeted by a chimeric single-guide RNA（sg RNA）. For plant genome editing, Agrobacterium-mediated T-DNA transformation has been broadly used to express Cas9 proteins and sg RNAs under the control of Ca MV 35 S and U6/U3 promoter, respectively. We here developed a simple and high-throughput binary vector system to clone a 19 20 bp of sg RNA, which binds to the reverse complement of a target locus, in a large T-DNA binary vector containing an Sp Cas9 expressing cassette. Twostep cloning procedures：（1） annealing two target-specific oligonucleotides with overhangs specific to the Aar I restriction enzyme site of the binary vector; and（2） ligating the annealed oligonucleotides into the two Aar I sites of the vector, facilitate the high-throughput production of the positive clones. In addition, Cas9-coding sequence and U6/U3 promoter can be easily exchanged via the GatewayTMsystem and unique Eco RI/Xho I sites on the vector, respectively. We examined the mutation ratio and patterns when we transformed these constructs into Arabidopsis thaliana and a wild tobacco, Nicotiana attenuata. Our vector system will be useful to generate targeted large-scale knock-out lines of model as well as non-model plant.展开更多
Objective To study the alteration of Na + Ca 2+ exchange in rat cardiac sarcolemmal membrane during phases of septic shock Methods Sepsis was induced by cecal ligation and puncture (CLP) Na + Ca 2+ ...Objective To study the alteration of Na + Ca 2+ exchange in rat cardiac sarcolemmal membrane during phases of septic shock Methods Sepsis was induced by cecal ligation and puncture (CLP) Na + Ca 2+ exchange was assayed by radioactive analysis Results Na + dependent 45 Ca 2+ uptake was decreased by 62%-69% in late phase of sepsis, whereas it was not affected in early phase of sepsis Na + Ca 2+ exchange stimulated by 5'guanylyl imidodiphosphate [Gpp(NH)p] was decreased by 65 7% in late phase of sepsis but unaltered in early phase of sepsis Two agonists (angiotensin Ⅱ and phenylephrine) coupled to Gq and a protein kinase C activator, phorbol 12 myristate 13 acetate (PMA) all inhibited Na + Ca 2+ exchange in late phase of sepsis Na + Ca 2+ exchange activities induced by phosphorylation of Na + Ca 2+ exchange were decreased in late phase of sepsis, whereas inhibition of Na + Ca 2+ exchange by dephosphorylation was increased both in early and late phases of sepsis Conclusion The alteration of Na + Ca 2+ exchange during different phases of sepsis might be related to the activities of Gq, protein kinase C, and phosphorylation/dephosphorylation展开更多
Changes in cellular Ca2+concentration control a variety of physiological activities including hormone and neurotransmitter release,muscular contraction,synaptic plas-ticity,ionic channel permeability,apoptosis,enzyme ...Changes in cellular Ca2+concentration control a variety of physiological activities including hormone and neurotransmitter release,muscular contraction,synaptic plas-ticity,ionic channel permeability,apoptosis,enzyme activity,gene transcription and reproduction process.Spatial-temporal Ca2+dynamics due to Ca2 t release,buffering and re-uptaking plays a central role in studying the Ca2+regulation in T lympho-cytes.In most cases,Ca2+has its major signaling function when it is elevated in the cytosolic compartment.In this paper,a two-dimensional mathematical model to study spatiotemporal variations of intracellular Ca2+concentration in T lymphocyte cell is proposed and investigated.The cell is assumed to be a circular shaped geomnetrical domain for the representation of properties of Ca2+dynamics within the cell includ-ing important parameters.Ca2+binding proteins for the dynamics of Ca2+are itself buffer and other physiological parameters located in Ca2+stores.The model incorpo-rates the important biophysical processes like difusion,reaction,voltage gated Ca2+channel,leak from endoplasmic reticulum(ER),efflux from cytosol to ER via sarco ER Ca2+adenosine triphosphate(SERCA)pumps,buffers and Na+/Ca2+exchanger.The proposed mathematical model is solved using a finite difference method and the finite element method.Appropriate initial and boundary conditions are incorporated in the model based on biophysical conditions of the problem.Computer simulations in MAT-LAB R2019b are employed to investigate mathematical models of reaction-diffusion equation.The effect of source,buffer,Nat/Ca2+exchanger,etc.on spatial and tempo-ral patterns of Ca2+in T lymphocyte has been studied with the help of numerical results.From the obtained results,it is observed that,the coordinated combination of the incor-porated parameters plays a significant role in Ca2+regulation in T lymphocytes.ER leak and voltage-gated Ca2+channel provides the necessary Ca2+to the cell when required for its proper functioning,while on the other side buffers,SERCA pump and Na+/Ca2+exchanger makes balance in the Ca2+concentration,so as to prevent the cell from death as higher concentration for longer time is harmful for the cell and can cause cell death.展开更多
文摘[Objective] A method for the determination of exchangeable Ca and Mg contents in tobacco growing area by atomic absorption spectrometry was established. [Method] Factors in the extraction process, sample weight, ammonium acetate volume, extraction time and strontium chloride volume were tested, and the stability, precision and accuracy of the method were also investigated. Meanwhile, samples at different pH values were determined by this method. [Result] The results showed that this method operation has simple operation with good stability and high precision. [Conclusion] This method could be applied to determine exchangeable Ca and Mg contents in tobacco growing area.
文摘A leaching experiment was carried out with repacked soil columns in laboratory to stndy the leachingprocess of a red soil derived from sandstone as affected by warions fertilization practices. The treatments wereCK (as a control), CaCO_3, CaSO_4, MgCO_3, Ca(H_2PO_4)_2, Urea, KCl, Mnltiple (a mixture of the above-mentioned fertilizers), and KNO_3. The fertilizers were added to the bare surface of the soil columns, andthen the columns were leached with 120 mL deionized water daily through peristaltic pumps over a periodof 92 days. At the end of leaching process, soils were sampled from different depths of the soil profiles, i.e.,0-5 cm, 5-10 cm, 10-20 cm, 20-40 cm, and 40-60 cm. The results showed when applying Ca, Mg, andK to the bare surface of the soil columns, exchangeable Ca￣(2+), Mg￣(2+), and K￣+ in the upper layer of thesoil profile increased correspondingly, with an extent depending mainly on the application rates of Ca, Mg,and K and showing a downward trend. CaCO_3, CaSO_4, MgCO_3, and Ca(H_2PO_4)_2 treatments had scarcelyany effect on movement of exchangeable K￣+, while CaCO_3, and CaSO_4 treatments significantly promotedthe downward movement of exchangeable Mg￣(2+) although these two treatments had no obvious effect onleaching losses of Mg. The fact that under Urea treatment, exchangeable Ca￣(2+) and Mg￣(2+) were higheras compared to CK treatment showed urea could prevent leaching of exchangeable Ca￣(2+) and Mg￣(2+). Theobvious downward movement of exchangeable Ca￣(2+) and Mg￣(2+) was noticed in KCl treatment. In Multipletreatment, the downward movement of exchangeable Ca￣(2+) and Mg￣(2+) was evident, while that of K￣+ was lessevident. Application of KNO_3 strongly promoted the downward movement of exchangeable Ca￣(2+) and Mg￣(2+)in the soil profile.
基金This work was supported by Leading Specialty Funding of Shanghai, Grant No.94- III- 001.
文摘To determine the effects of HOE 694, a new and potent Na＋- H＋ exchanger blocker, on transient inward current (Iti) and Na＋- Ca2＋ exchange during hypoxia- reoxygenation in guinea pig cardiomyocytes. Methods. Cardiomyocytes were isolated from adult guinea pig ventricle. Experiment was performed in an experimental chamber that allowed the cells to be exposed to a sufficiently low O2 pressure. The cells were subjected to hypoxia and reoxygenation. The ionic currents were studied with patch clamp technique. Results. In the absence of HOE 694, hypoxia- reoxygenation induced Iti in 12 of 15 experiments; but in cardiomyocytes pretreated with HOE 694 (10～ 50μ mol/L), the incidence of Iti observed during reoxygenation was reduced to 5 of 11 experiments and 3 of 10 experiments, P Conclusions. Blockade of the Na＋- H＋ exchange by HOE 694 could reduce Ca2＋ overload upon hypoxia- reoxygenation, and inhibition of Na＋- H＋ exchange may also indirectly decrease Na＋- Ca2＋ exchange activity during hypoxia.
基金The authors acknowledge the support of the National Natural Scicnce Foundation of ChinaProvincial Natural Science Foundation of Shanxi.
文摘We determined whether La3+ enter human peripheral blood lymphocytes via Na+/Ca2+ exchanger (measured with fura-2). We first compared the sensitivity of fura-2 with La3+ and Ca2+, the result indicates that the sensitivity of fura-2 for La3+ is much greater than for Ca2+. La3+ forms a 1:1 La3+-fura-2 complex (apparent dissociation constant = 1.7x10(-12) mol/L, pH 7.05). Ouabain-pretreated cells, suspended in Na+-free medium, showed that La3+ can enter human lymphocytes via the Na-i(+)/Ca2+ (La3+)(o) exchanger and is found to be about 10(-12) mol/L in cells exposed to 0.4 mmol/L La3+. Otherwise, the higher concentration (0.1 mmol/L) blocks the Na-i(+)/Ca2+(La3+)(o) exchange-mediated influx of Ca2+, but the lower concentration (0.01 mmol/L) appears to increase Ca2+ entry.
文摘At present, there are relevant scientific materials on the cellular and molecular mechanisms of electrogenic Na/K pump function and structure, as well as on the potential- and ligand-activated ionic channels in the membrane. However, the role of electrogenic Na/K pump in regulation of semipermeable properties of cell membrane has not been elucidated yet, which is due to the fact that our knowledge about the biophysical properties of cell membrane is based on the conductive membrane theory of Hodgkin-Huxley-Katz, which is developed on internally perfused squid axon and lacks intracellular metabolism. Thus, the accumulated abundance of data on the role of G-proteins-dependent intracellular signaling system in regulation of Na/K pump activity and biophysical properties of cell membrane presumes fundamental revision of some statements of membrane theory. The aim of the present review is to briefly demonstrate our and literature data on cell hydration-induced auto-regulation of Na/K pump as well as on its role in metabolic control of semipermeable properties and excitability of neuronal membrane, which are omitted in the study of internally perfused squid axon.
基金supported by Institute for Basic Science (IBS-R021-D1)
文摘CRISPR-Cas9 system is now widely used to edit a target genome in animals and plants. Cas9 protein derived from Streptococcus pyogenes（Sp Cas9） cleaves double-stranded DNA targeted by a chimeric single-guide RNA（sg RNA）. For plant genome editing, Agrobacterium-mediated T-DNA transformation has been broadly used to express Cas9 proteins and sg RNAs under the control of Ca MV 35 S and U6/U3 promoter, respectively. We here developed a simple and high-throughput binary vector system to clone a 19 20 bp of sg RNA, which binds to the reverse complement of a target locus, in a large T-DNA binary vector containing an Sp Cas9 expressing cassette. Twostep cloning procedures：（1） annealing two target-specific oligonucleotides with overhangs specific to the Aar I restriction enzyme site of the binary vector; and（2） ligating the annealed oligonucleotides into the two Aar I sites of the vector, facilitate the high-throughput production of the positive clones. In addition, Cas9-coding sequence and U6/U3 promoter can be easily exchanged via the GatewayTMsystem and unique Eco RI/Xho I sites on the vector, respectively. We examined the mutation ratio and patterns when we transformed these constructs into Arabidopsis thaliana and a wild tobacco, Nicotiana attenuata. Our vector system will be useful to generate targeted large-scale knock-out lines of model as well as non-model plant.
文摘Objective To study the alteration of Na + Ca 2+ exchange in rat cardiac sarcolemmal membrane during phases of septic shock Methods Sepsis was induced by cecal ligation and puncture (CLP) Na + Ca 2+ exchange was assayed by radioactive analysis Results Na + dependent 45 Ca 2+ uptake was decreased by 62%-69% in late phase of sepsis, whereas it was not affected in early phase of sepsis Na + Ca 2+ exchange stimulated by 5'guanylyl imidodiphosphate [Gpp(NH)p] was decreased by 65 7% in late phase of sepsis but unaltered in early phase of sepsis Two agonists (angiotensin Ⅱ and phenylephrine) coupled to Gq and a protein kinase C activator, phorbol 12 myristate 13 acetate (PMA) all inhibited Na + Ca 2+ exchange in late phase of sepsis Na + Ca 2+ exchange activities induced by phosphorylation of Na + Ca 2+ exchange were decreased in late phase of sepsis, whereas inhibition of Na + Ca 2+ exchange by dephosphorylation was increased both in early and late phases of sepsis Conclusion The alteration of Na + Ca 2+ exchange during different phases of sepsis might be related to the activities of Gq, protein kinase C, and phosphorylation/dephosphorylation
基金Author of the paper is very grateful to Xi'an Jiaotong University for the postdoc-toral position provided to him.Also,the author would like to thank the reviewers and editors of this feedback that improved the presentation of the paper greatlyThis study was supported by the Grant from China Postdoctoral Science Foundation(Grant No.2019M663653)The funding body did not play any roles in the design of the study and in writing this paper.
文摘Changes in cellular Ca2+concentration control a variety of physiological activities including hormone and neurotransmitter release,muscular contraction,synaptic plas-ticity,ionic channel permeability,apoptosis,enzyme activity,gene transcription and reproduction process.Spatial-temporal Ca2+dynamics due to Ca2 t release,buffering and re-uptaking plays a central role in studying the Ca2+regulation in T lympho-cytes.In most cases,Ca2+has its major signaling function when it is elevated in the cytosolic compartment.In this paper,a two-dimensional mathematical model to study spatiotemporal variations of intracellular Ca2+concentration in T lymphocyte cell is proposed and investigated.The cell is assumed to be a circular shaped geomnetrical domain for the representation of properties of Ca2+dynamics within the cell includ-ing important parameters.Ca2+binding proteins for the dynamics of Ca2+are itself buffer and other physiological parameters located in Ca2+stores.The model incorpo-rates the important biophysical processes like difusion,reaction,voltage gated Ca2+channel,leak from endoplasmic reticulum(ER),efflux from cytosol to ER via sarco ER Ca2+adenosine triphosphate(SERCA)pumps,buffers and Na+/Ca2+exchanger.The proposed mathematical model is solved using a finite difference method and the finite element method.Appropriate initial and boundary conditions are incorporated in the model based on biophysical conditions of the problem.Computer simulations in MAT-LAB R2019b are employed to investigate mathematical models of reaction-diffusion equation.The effect of source,buffer,Nat/Ca2+exchanger,etc.on spatial and tempo-ral patterns of Ca2+in T lymphocyte has been studied with the help of numerical results.From the obtained results,it is observed that,the coordinated combination of the incor-porated parameters plays a significant role in Ca2+regulation in T lymphocytes.ER leak and voltage-gated Ca2+channel provides the necessary Ca2+to the cell when required for its proper functioning,while on the other side buffers,SERCA pump and Na+/Ca2+exchanger makes balance in the Ca2+concentration,so as to prevent the cell from death as higher concentration for longer time is harmful for the cell and can cause cell death.
