Protein extraction is a critical step for two-dimensional electrophoresis （2-DE）. Different plant samples require different and adaptive protein extraction protocols. The leaves of medicinal plant, Baphicacanthus cu...Protein extraction is a critical step for two-dimensional electrophoresis （2-DE）. Different plant samples require different and adaptive protein extraction protocols. The leaves of medicinal plant, Baphicacanthus cusia （Nees） Bremek are notoriously recalcitrant to common protein extraction methods due to high levels of interfering compounds （especially the secondary metabolites and pigments）. This study was aimed to establish a routine procedure for the proteomic analysis ofB. cusia leaves, and a new protocol for the protein extraction was developed by optimizing trichloroacetic acid （TCA）/ acetone extraction method. The efficiency of this protocol was demonstrated by comparison with 3 published protein extraction methods （chloroform/acetone, Mg/NP-40, Tris-base/acetone）. The results showed that the optimized TCA/ acetone precipitation extraction method gave a relatively high protein yield （9.263 mg g^-1 fresh weight）, high-resolution separation, clear protein profiles, the highest proteins spots （1 31 t protein spots）, and displayed less contamination in 2- DE gels. Therefore, the results suggested that the optimized TCA/acetone method was the most effective among the 4 methods for B. cusia leaves.展开更多
The aim of this experiment is to study the mechanism of APN in alleviating the Ca2+-overloading in dog model during the process of ischemic reperfusion. In comparison with the sustained ischemic group, the parameters ...The aim of this experiment is to study the mechanism of APN in alleviating the Ca2+-overloading in dog model during the process of ischemic reperfusion. In comparison with the sustained ischemic group, the parameters in the ischemic reperfusion group demonstrated: Ca2+ of ischemic region of myocardial cell increased (P<0.05) , Na+ increased remarkably(P<0. 01), the activity of Ca2+-ATPase dropped remarkably(P<0. 01),and MDA increased significantly(P<0. 01). Whereas in the group pretreated with APN, the Ca2+ in the relevant area reduced (P<0.05), Na+ decreased significantly(P<0. 01), the activity of Ca2+-ATPase and Na+-K+ ATPase increased remarkably(P<0.01) ,and MDA decreased significantly (P<0. 01). These findings indicate that APN may improve the activity of sarcolemma ATPase in alleviating the Ca2+ and Na+-overloading by decreasing the harmful effect of oxygen free radicals.展开更多
In experimental dogs,the effect of APN in alleviating the ischemiareperfusion injury was prominent.Compared with the sustained isthmian group,superoxide dismutast(SOD)in the ischemic region of myocardial tissue in the...In experimental dogs,the effect of APN in alleviating the ischemiareperfusion injury was prominent.Compared with the sustained isthmian group,superoxide dismutast(SOD)in the ischemic region of myocardial tissue in the ischemiareperfusion group was significantly decreased and malondialdehyde(MDA)marktdly increased Ca2+in myocardial cells was increased. and ultrastructural changes of myocardial tisstes were severe in the APN-pretreated ischemia-reperfusion group ,on the contrary,all the above parameters showed reversely,i,e.,SOD increased ,MDA and intracellular Ca2+ decreased.the ultrastructure changes were less distorted.展开更多
Machilus spp.are mostly valuable timber and economic tree species,widely used in many fields such as artificial afforestation,landscaping,furniture and building materials,chemicals and medicine.In this article,the dis...Machilus spp.are mostly valuable timber and economic tree species,widely used in many fields such as artificial afforestation,landscaping,furniture and building materials,chemicals and medicine.In this article,the distribution,uses and domestic resources of Machilus Nees are summarized.Focus is placed on the analysis of propagation methods,including seed propagation,cutting propagation and tissue culture propagation.The research status and existing problems are summarized,and the future research focus on the propagation technology of Machilus Nees is prospected,with a view to provide theoretical references for the efficient breeding of high-quality seedlings and comprehensive promotion and application of Machilus spp.and contribute to the conservation and sustainable use of wild resources.展开更多
[Objectives]To observe the effects of polysaccharides from Dicliptera chinensis(L.)Nees.on the expression of TLR/NF-κB pathway related proteins in HepG2 cells induced by oleic acid,and to explore the possible mechani...[Objectives]To observe the effects of polysaccharides from Dicliptera chinensis(L.)Nees.on the expression of TLR/NF-κB pathway related proteins in HepG2 cells induced by oleic acid,and to explore the possible mechanism of polysaccharides from D.chinensis(L.)Nees.in the treatment of non-alcoholic fatty liver disease(NAFLD).[Methods]HepG2 cells were induced with oleic acid to establish a non-alcoholic fatty liver cell model.After intervention with 0.25 and 0.5 mg/mL of D.chinensis(L.)Nees.polysaccharides,the ALT and AST activity and TG and TC contents were detected with kits,and the changes in the expression of CDK5,TLR4,p-NF-κB and NF-κB were analyzed using Western-blotting.[Results]In the HepG2 cells induced with oleic acid,the ALT and AST activity increased significantly,the TG and TC contents increased significantly,and the expression levels of CDK5,TLR4 and p-NF-κB proteins up-regulated significantly.In the HepG2 cells intervened with D.chinensis(L.)Nees.polysaccharides,the activity of ALT and AST,the contents of TG and TC,and the expression levels of CDK5,TLR4 and p-NF-κB proteins all reduced significantly.[Conclusions]Polysaccharides from D.chinensis(L.)Nees.may interfere with NAFLD by inhibiting the TLR4/NF-κB pathway.展开更多
[Objective] The study aimed to establish a protoplast transformation system in Alternaria tenuissima. [Method] The protoplast of A.tenuissima was firstly prepared by enzymolysis method; then the yielded protoplast was...[Objective] The study aimed to establish a protoplast transformation system in Alternaria tenuissima. [Method] The protoplast of A.tenuissima was firstly prepared by enzymolysis method; then the yielded protoplast was transformed by G418 resistant DNA plasmid using PEG/CaCl2 method. [Result] The growth phenotype and PCR detection showed that resistance gene had integrated into A.tenuissima genome. The transformation efficiency of this method reached per μg DNA 3-4 transformants. After subculture thrice under nonselective condition, G418 resistance could still inherit stably. [Conclusion] The transformation system of A.tenuissima was successfully established, which laid basis for studying of the gene function of Alternaria tenuissima.展开更多
基金supported by the Department of Edu-cation(JA05238)the Key Project on Ecology of Fujian Province,China(0608537)
文摘Protein extraction is a critical step for two-dimensional electrophoresis （2-DE）. Different plant samples require different and adaptive protein extraction protocols. The leaves of medicinal plant, Baphicacanthus cusia （Nees） Bremek are notoriously recalcitrant to common protein extraction methods due to high levels of interfering compounds （especially the secondary metabolites and pigments）. This study was aimed to establish a routine procedure for the proteomic analysis ofB. cusia leaves, and a new protocol for the protein extraction was developed by optimizing trichloroacetic acid （TCA）/ acetone extraction method. The efficiency of this protocol was demonstrated by comparison with 3 published protein extraction methods （chloroform/acetone, Mg/NP-40, Tris-base/acetone）. The results showed that the optimized TCA/ acetone precipitation extraction method gave a relatively high protein yield （9.263 mg g^-1 fresh weight）, high-resolution separation, clear protein profiles, the highest proteins spots （1 31 t protein spots）, and displayed less contamination in 2- DE gels. Therefore, the results suggested that the optimized TCA/acetone method was the most effective among the 4 methods for B. cusia leaves.
文摘The aim of this experiment is to study the mechanism of APN in alleviating the Ca2+-overloading in dog model during the process of ischemic reperfusion. In comparison with the sustained ischemic group, the parameters in the ischemic reperfusion group demonstrated: Ca2+ of ischemic region of myocardial cell increased (P<0.05) , Na+ increased remarkably(P<0. 01), the activity of Ca2+-ATPase dropped remarkably(P<0. 01),and MDA increased significantly(P<0. 01). Whereas in the group pretreated with APN, the Ca2+ in the relevant area reduced (P<0.05), Na+ decreased significantly(P<0. 01), the activity of Ca2+-ATPase and Na+-K+ ATPase increased remarkably(P<0.01) ,and MDA decreased significantly (P<0. 01). These findings indicate that APN may improve the activity of sarcolemma ATPase in alleviating the Ca2+ and Na+-overloading by decreasing the harmful effect of oxygen free radicals.
文摘In experimental dogs,the effect of APN in alleviating the ischemiareperfusion injury was prominent.Compared with the sustained isthmian group,superoxide dismutast(SOD)in the ischemic region of myocardial tissue in the ischemiareperfusion group was significantly decreased and malondialdehyde(MDA)marktdly increased Ca2+in myocardial cells was increased. and ultrastructural changes of myocardial tisstes were severe in the APN-pretreated ischemia-reperfusion group ,on the contrary,all the above parameters showed reversely,i,e.,SOD increased ,MDA and intracellular Ca2+ decreased.the ultrastructure changes were less distorted.
基金Forestry Science and Technology Innovation Project of Guangdong Province(14KJCX005-02).
文摘Machilus spp.are mostly valuable timber and economic tree species,widely used in many fields such as artificial afforestation,landscaping,furniture and building materials,chemicals and medicine.In this article,the distribution,uses and domestic resources of Machilus Nees are summarized.Focus is placed on the analysis of propagation methods,including seed propagation,cutting propagation and tissue culture propagation.The research status and existing problems are summarized,and the future research focus on the propagation technology of Machilus Nees is prospected,with a view to provide theoretical references for the efficient breeding of high-quality seedlings and comprehensive promotion and application of Machilus spp.and contribute to the conservation and sustainable use of wild resources.
基金National Natural Science Foundation of China(81960779,81760114,81660104,81860673)Natural Science Foundation of Guangxi Zhuang Autonomous Region(2017GXNSFAA198218,2017GXNSFAA198326,2018GXNSFAA281040)。
文摘[Objectives]To observe the effects of polysaccharides from Dicliptera chinensis(L.)Nees.on the expression of TLR/NF-κB pathway related proteins in HepG2 cells induced by oleic acid,and to explore the possible mechanism of polysaccharides from D.chinensis(L.)Nees.in the treatment of non-alcoholic fatty liver disease(NAFLD).[Methods]HepG2 cells were induced with oleic acid to establish a non-alcoholic fatty liver cell model.After intervention with 0.25 and 0.5 mg/mL of D.chinensis(L.)Nees.polysaccharides,the ALT and AST activity and TG and TC contents were detected with kits,and the changes in the expression of CDK5,TLR4,p-NF-κB and NF-κB were analyzed using Western-blotting.[Results]In the HepG2 cells induced with oleic acid,the ALT and AST activity increased significantly,the TG and TC contents increased significantly,and the expression levels of CDK5,TLR4 and p-NF-κB proteins up-regulated significantly.In the HepG2 cells intervened with D.chinensis(L.)Nees.polysaccharides,the activity of ALT and AST,the contents of TG and TC,and the expression levels of CDK5,TLR4 and p-NF-κB proteins all reduced significantly.[Conclusions]Polysaccharides from D.chinensis(L.)Nees.may interfere with NAFLD by inhibiting the TLR4/NF-κB pathway.
文摘[Objective] The study aimed to establish a protoplast transformation system in Alternaria tenuissima. [Method] The protoplast of A.tenuissima was firstly prepared by enzymolysis method; then the yielded protoplast was transformed by G418 resistant DNA plasmid using PEG/CaCl2 method. [Result] The growth phenotype and PCR detection showed that resistance gene had integrated into A.tenuissima genome. The transformation efficiency of this method reached per μg DNA 3-4 transformants. After subculture thrice under nonselective condition, G418 resistance could still inherit stably. [Conclusion] The transformation system of A.tenuissima was successfully established, which laid basis for studying of the gene function of Alternaria tenuissima.
