Insulin-like growth factor 1 （IGF-I） is a potential nutrient for nerve repair. However, it is impractical as a therapy because of its limited half- life, rapid clearance, and limited target specificity. To achieve t...Insulin-like growth factor 1 （IGF-I） is a potential nutrient for nerve repair. However, it is impractical as a therapy because of its limited half- life, rapid clearance, and limited target specificity. To achieve targeted and long-lasting treatment, we investigated the addition of a binding structure by fusing a collagen-binding domain to IGF- 1. After confirming its affinity for collagen, the biological activity of this construct was examined by measuring cell proliferation after transfection into PC12 and Schwann cells using a 3-（4,5-dimethyl-2-thiazolyl）-2,5-di- phenyl-2-H-tetrazolium bromide assay. Immunofluorescence staining was conducted to detect neurofilament and microtubule-associated protein 2 expression, while real time-polymerase chain reaction was utilized to determine IGF-1 receptor and nerve growth/actor mRNA expression. Our results demonstrate a significant increase in collagen-binding activity of the recombinant protein compared with IGF-1. Moreover, the recombinant protein promoted proliferation of PC12 and Schwann cells, and increased the expression of neurofilament and microtubule-associated protein 2. Importantly, the recombinant protein also stimulated sustained expression of IGF-1 receptor and nerve growth factor mRNA for days. These results show that the recombinant protein achieved the goal of targeting and long-lasting treatment, and thus could become a clinically used factor for promoting nerve regeneration with a prolonged therapeutic effect.展开更多
In an earlier paper, the author employed the thesis that baryons are Yang-Mills magnetic monopoles and that proton and neutron binding energies are determined based on their up and down current quark masses to predict...In an earlier paper, the author employed the thesis that baryons are Yang-Mills magnetic monopoles and that proton and neutron binding energies are determined based on their up and down current quark masses to predict a relationship among the electron and up and down quark masses within experimental errors and to obtain a very accurate relationship for nuclear binding energies generally and for the binding of 56Fe in particular. The free proton and neutron were understood to each contain intrinsic binding energies which confine their quarks, wherein some or most (never all) of this energy is released for binding when they are fused into composite nuclides. The purpose of this paper is to further advance this thesis by seeing whether it can explain the specific empirical binding energies of the light 1s nuclides, namely, 2H, 3H, 3He and 4He, with high precision. As the method to achieve this, we show how these 1s binding energies are in fact the components of inner and outer tensor products of Yang-Mills matrices which are implicit in the expressions for these intrinsic binding energies. The result is that the binding energies for the 4He, 3He and 3H nucleons are respectively, independently, explained to less than four parts in one million, four parts in 100,000, and seven parts in one million, all in AMU. Further, we are able to exactly relate the neutron minus proton mass difference to a function of the up and down current quark masses, which in turn enables us to explain the 2H binding energy most precisely of all, to just over 8 parts in ten million. These energies have never before been theoretically explained with such accuracy, which leads to the conclusion that the underlying thesis provides the strongest theoretical explanation to date of what baryons are, and of how protons and neutrons confine their quarks and bind together into composite nuclides. As is also reviewed in Section 9, these results may lay the foundation for more easily catalyzing nuclear fusion energy release.展开更多
With the total RNA of Dunaliella salina as a template,the cD NA sequence of D. salina small GTP-binding protein gene was amplified by RT-PCR technique,and cloned onto pM Dl8-T simple vector,the recon was subjected to ...With the total RNA of Dunaliella salina as a template,the cD NA sequence of D. salina small GTP-binding protein gene was amplified by RT-PCR technique,and cloned onto pM Dl8-T simple vector,the recon was subjected to PCR detection and restriction endonuclease analysis,and the total sequence of DNA was determined. The results showed that the cloned fragment was 612 bp,and shared 100% homology with reported D. salina DsRab gene（ GenB ank： JN989548）. The target gene fragment was inserted downstream of pM DCG 35 S promoter,constructing subcellular localization recombinant vector pM DCG-DsRab. The successfully constructed subcellular localization recombinant vector pM DCG-DsRab was transformed into Agrobacterium tumefaciens LBA4404,and positive single clones were screened and used for transinfection of onion epidemical cells by Agrobacterium-mediated method,and the instant expression of DsRab was observed under fluorescence microscope. The results showed that the fusion protein GFP-DsRab was successfully expressed in onion epidemical cells,and mainly distributed on cytomembrane. This study will provide reference for further illumination of the function and action mechanism of D. salina small GTP-binding protein DsRab.展开更多
基金supported by the National Natural Science Foundation of China,No.81350013a grant from the Jilin Provincial Science and Technology Plan of China,No.20160101027JC&SC201502001the Graduate Innovation Fund of Jilin University in China,No.2017031&2017176
文摘Insulin-like growth factor 1 （IGF-I） is a potential nutrient for nerve repair. However, it is impractical as a therapy because of its limited half- life, rapid clearance, and limited target specificity. To achieve targeted and long-lasting treatment, we investigated the addition of a binding structure by fusing a collagen-binding domain to IGF- 1. After confirming its affinity for collagen, the biological activity of this construct was examined by measuring cell proliferation after transfection into PC12 and Schwann cells using a 3-（4,5-dimethyl-2-thiazolyl）-2,5-di- phenyl-2-H-tetrazolium bromide assay. Immunofluorescence staining was conducted to detect neurofilament and microtubule-associated protein 2 expression, while real time-polymerase chain reaction was utilized to determine IGF-1 receptor and nerve growth/actor mRNA expression. Our results demonstrate a significant increase in collagen-binding activity of the recombinant protein compared with IGF-1. Moreover, the recombinant protein promoted proliferation of PC12 and Schwann cells, and increased the expression of neurofilament and microtubule-associated protein 2. Importantly, the recombinant protein also stimulated sustained expression of IGF-1 receptor and nerve growth factor mRNA for days. These results show that the recombinant protein achieved the goal of targeting and long-lasting treatment, and thus could become a clinically used factor for promoting nerve regeneration with a prolonged therapeutic effect.
文摘In an earlier paper, the author employed the thesis that baryons are Yang-Mills magnetic monopoles and that proton and neutron binding energies are determined based on their up and down current quark masses to predict a relationship among the electron and up and down quark masses within experimental errors and to obtain a very accurate relationship for nuclear binding energies generally and for the binding of 56Fe in particular. The free proton and neutron were understood to each contain intrinsic binding energies which confine their quarks, wherein some or most (never all) of this energy is released for binding when they are fused into composite nuclides. The purpose of this paper is to further advance this thesis by seeing whether it can explain the specific empirical binding energies of the light 1s nuclides, namely, 2H, 3H, 3He and 4He, with high precision. As the method to achieve this, we show how these 1s binding energies are in fact the components of inner and outer tensor products of Yang-Mills matrices which are implicit in the expressions for these intrinsic binding energies. The result is that the binding energies for the 4He, 3He and 3H nucleons are respectively, independently, explained to less than four parts in one million, four parts in 100,000, and seven parts in one million, all in AMU. Further, we are able to exactly relate the neutron minus proton mass difference to a function of the up and down current quark masses, which in turn enables us to explain the 2H binding energy most precisely of all, to just over 8 parts in ten million. These energies have never before been theoretically explained with such accuracy, which leads to the conclusion that the underlying thesis provides the strongest theoretical explanation to date of what baryons are, and of how protons and neutrons confine their quarks and bind together into composite nuclides. As is also reviewed in Section 9, these results may lay the foundation for more easily catalyzing nuclear fusion energy release.
基金Supported by National Natural Science Foundation of China(31472260)
文摘With the total RNA of Dunaliella salina as a template,the cD NA sequence of D. salina small GTP-binding protein gene was amplified by RT-PCR technique,and cloned onto pM Dl8-T simple vector,the recon was subjected to PCR detection and restriction endonuclease analysis,and the total sequence of DNA was determined. The results showed that the cloned fragment was 612 bp,and shared 100% homology with reported D. salina DsRab gene（ GenB ank： JN989548）. The target gene fragment was inserted downstream of pM DCG 35 S promoter,constructing subcellular localization recombinant vector pM DCG-DsRab. The successfully constructed subcellular localization recombinant vector pM DCG-DsRab was transformed into Agrobacterium tumefaciens LBA4404,and positive single clones were screened and used for transinfection of onion epidemical cells by Agrobacterium-mediated method,and the instant expression of DsRab was observed under fluorescence microscope. The results showed that the fusion protein GFP-DsRab was successfully expressed in onion epidemical cells,and mainly distributed on cytomembrane. This study will provide reference for further illumination of the function and action mechanism of D. salina small GTP-binding protein DsRab.
