The cholinergic anti-inflammatory pathway （CAP） is a neurophysiological mechanism that regulates the immune system. The CAP inhibits inflammation by suppressing cytokine synthesis via release of acetylcholine in org...The cholinergic anti-inflammatory pathway （CAP） is a neurophysiological mechanism that regulates the immune system. The CAP inhibits inflammation by suppressing cytokine synthesis via release of acetylcholine in organs of the reticuloendothelial system, including the lungs, spleen, liver, kidneys and gastrointestinal tract. Acetylcholine can interact with a 7 nicotinic acetylcholine receptors （ a 7 nAchR） expressed by macrophages and other cytokine producing cells, down-regulate pro-inflammatory cytokine synthesis and prevent tissue damage. Herein is a review of the neurophysiological mechanism in which the CAP regulates inflammatory response, as well as its potential interventional strategy for inflammatory diseases.展开更多
The highly specific ligand of the N-acetylcholine receptor（N-AChR）,alpha-bungarotoxin,was used to determine the effect of soman,sarin and VX onN-AChR of the diaphragm and extensor digitorum Iongus muscle of mice and...The highly specific ligand of the N-acetylcholine receptor（N-AChR）,alpha-bungarotoxin,was used to determine the effect of soman,sarin and VX onN-AChR of the diaphragm and extensor digitorum Iongus muscle of mice andrats.The effects of the three anti-cholinesterase agents on N-AChR weredifferent.Sarin did not act directly on N-AChR and cause a change in the numberof N-AChR.VX decreased the binding sites of the receptor by binding with N-AChRdirectly.The LD<sub>50</sub>was 0.054mg/kg in mousse.Soman increased the binding sites,e.g.1～1.5 LD<sub>50</sub>soman increased the number of N-AChR of mouse diaphragm by 25%.The peak increaseof N-AChR was reached 0.5 h after poisoning and could last 96h.The receptornumber was still 22% higher than that of the control on the fourth day aftersoman poisoning in rats.Soman mainly increased the number of extrasynapticN-AChR,leading to the enhancement of sensitivity of cholinergic effectors toacetylcholine（ACh）,which is similar to the hypersensitiveness resulting fromdenervation.These findings are of significance in probing the receptor mecha-nisms and treatment of soman poisoning.展开更多
[ Objective ] This paper aimed to obtain active alpha-bungarotoxin （α-BGT） protein and investigate the isolation and purification of recombinant α-BGT protein. [ Method] The expressed soluble fusion protein was pu...[ Objective ] This paper aimed to obtain active alpha-bungarotoxin （α-BGT） protein and investigate the isolation and purification of recombinant α-BGT protein. [ Method] The expressed soluble fusion protein was purified by using GSTrap FF affinity columns. Purified fusion protein bound to GSTrap FF affinity col- umns was directly cleaved by thrombin to obtain the solution containing recombinant α-BGT. Using natural α-BGT as control, the antigenicity and biological activity of the purified fusion protein and recombinant α-BGT were detected by SDS-PAGE, Western Blot and toxicity test in vivo. [ Result] Recombinant ot-BGT was iso- lated; the purified fusion protein and recombinant α-BGT were similar to the natural α-BGT in antigenicity; the toxicity of purified fusion protein was relatively wea- ker; recombinant α-BGT was similar to the natural ot-BGT in toxicity. [Condusion] This study laid the foundation for further large-scale production of recombi- nant α-BGT.展开更多
[Objective] This study aimed to obtain recombinant alpha-bungarotoxin （a-BG-0 gene fusion protein with biological activity and investiagte its fusion expression. [Method] The plasmid pGEX-a-BGT was transformed into E...[Objective] This study aimed to obtain recombinant alpha-bungarotoxin （a-BG-0 gene fusion protein with biological activity and investiagte its fusion expression. [Method] The plasmid pGEX-a-BGT was transformed into E coil BL21 （DE3） and BL21 （DE3） plysS host bacteria to identify the optimal engineering strain. Fusion expression of the optimal engineering strain was induced, in order to optimize the induced expression conditions of the soluble fusion protein. [Result] JP-a-BGT was identified as the optimal engineering strain, which could express fusion protein after induced by IPTG. The optimal induced expression conditions of the soluble fusion protein were investigatect JP-a-BGT was incubated at 37 ℃ for 2.5 h and induced with 0.50 mmol4. IPTG for 4 h at 22 ℃, and the expression level of the soluble fusion protein reached 18.42%. [Conclusion] This study laid a solid foundation for the subsequent purification of fusion proteins and the separation and purification of a-BGT.展开更多
基金This study was supported by grants from the National Natural Science Foundation of China （No. 30672178, 30872683, 30800437） and the National Basic Research Program of China （No. 2005CB522602）.
文摘The cholinergic anti-inflammatory pathway （CAP） is a neurophysiological mechanism that regulates the immune system. The CAP inhibits inflammation by suppressing cytokine synthesis via release of acetylcholine in organs of the reticuloendothelial system, including the lungs, spleen, liver, kidneys and gastrointestinal tract. Acetylcholine can interact with a 7 nicotinic acetylcholine receptors （ a 7 nAchR） expressed by macrophages and other cytokine producing cells, down-regulate pro-inflammatory cytokine synthesis and prevent tissue damage. Herein is a review of the neurophysiological mechanism in which the CAP regulates inflammatory response, as well as its potential interventional strategy for inflammatory diseases.
文摘The highly specific ligand of the N-acetylcholine receptor（N-AChR）,alpha-bungarotoxin,was used to determine the effect of soman,sarin and VX onN-AChR of the diaphragm and extensor digitorum Iongus muscle of mice andrats.The effects of the three anti-cholinesterase agents on N-AChR weredifferent.Sarin did not act directly on N-AChR and cause a change in the numberof N-AChR.VX decreased the binding sites of the receptor by binding with N-AChRdirectly.The LD<sub>50</sub>was 0.054mg/kg in mousse.Soman increased the binding sites,e.g.1～1.5 LD<sub>50</sub>soman increased the number of N-AChR of mouse diaphragm by 25%.The peak increaseof N-AChR was reached 0.5 h after poisoning and could last 96h.The receptornumber was still 22% higher than that of the control on the fourth day aftersoman poisoning in rats.Soman mainly increased the number of extrasynapticN-AChR,leading to the enhancement of sensitivity of cholinergic effectors toacetylcholine（ACh）,which is similar to the hypersensitiveness resulting fromdenervation.These findings are of significance in probing the receptor mecha-nisms and treatment of soman poisoning.
文摘[ Objective ] This paper aimed to obtain active alpha-bungarotoxin （α-BGT） protein and investigate the isolation and purification of recombinant α-BGT protein. [ Method] The expressed soluble fusion protein was purified by using GSTrap FF affinity columns. Purified fusion protein bound to GSTrap FF affinity col- umns was directly cleaved by thrombin to obtain the solution containing recombinant α-BGT. Using natural α-BGT as control, the antigenicity and biological activity of the purified fusion protein and recombinant α-BGT were detected by SDS-PAGE, Western Blot and toxicity test in vivo. [ Result] Recombinant ot-BGT was iso- lated; the purified fusion protein and recombinant α-BGT were similar to the natural α-BGT in antigenicity; the toxicity of purified fusion protein was relatively wea- ker; recombinant α-BGT was similar to the natural ot-BGT in toxicity. [Condusion] This study laid the foundation for further large-scale production of recombi- nant α-BGT.
文摘[Objective] This study aimed to obtain recombinant alpha-bungarotoxin （a-BG-0 gene fusion protein with biological activity and investiagte its fusion expression. [Method] The plasmid pGEX-a-BGT was transformed into E coil BL21 （DE3） and BL21 （DE3） plysS host bacteria to identify the optimal engineering strain. Fusion expression of the optimal engineering strain was induced, in order to optimize the induced expression conditions of the soluble fusion protein. [Result] JP-a-BGT was identified as the optimal engineering strain, which could express fusion protein after induced by IPTG. The optimal induced expression conditions of the soluble fusion protein were investigatect JP-a-BGT was incubated at 37 ℃ for 2.5 h and induced with 0.50 mmol4. IPTG for 4 h at 22 ℃, and the expression level of the soluble fusion protein reached 18.42%. [Conclusion] This study laid a solid foundation for the subsequent purification of fusion proteins and the separation and purification of a-BGT.
