Genetic manipulation(either restraint or enhancement)of the biosynthesis pathway ofα-linolenic acid(ALA)in seed oil is an important goal in Brassica napus breeding.B.napus is a tetraploid plant whose genome often har...Genetic manipulation(either restraint or enhancement)of the biosynthesis pathway ofα-linolenic acid(ALA)in seed oil is an important goal in Brassica napus breeding.B.napus is a tetraploid plant whose genome often har-bors four and six homologous copies,respectively,of the two fatty acid desaturases FAD2 and FAD3,which con-trol the last two steps of ALA biosynthesis during seed oil accumulation.In this study,we compared their promoters,coding sequences,and expression levels in three high-ALA inbred lines 2006L,R8Q10,and YH25005,a low-ALA line A28,a low-ALA/high-oleic-acid accession SW,and the wildtype ZS11.The expression levels of most FAD2 and FAD3 homologs in the three high-ALA accessions were higher than those in ZS11 and much higher than those in A28 and SW.The three high-ALA accessions shared similar sequences with the pro-moters and CDSs of BnFAD3.C4 and BnFAD3.A3.In A28 and SW,substitution of three amino acid residues in BnFAD2.A5 and BnFAD2.C5,an absence of BnFAD2.C1 locus,and a 549 bp long deletion on the BnFAD3.A3 promoter were detected.The profile of BnFAD2 mutation in the two low-ALA accessions A28 and SW is different from that reported in previous studies.The mutations in BnFAD3 in the high-ALA accessions are reported for thefirst time.In identifying the sites of these mutations,we provide detailed information to aid the design of mole-cular markers for accelerated breeding schemes.展开更多
5×FAD转基因小鼠(transgenic mice with five familial Alzheimer’s disease)是携带5个家族性基因突变的APP/PS1转基因小鼠,其中与β-淀粉样蛋白前体(amyloid precursor protein,APP)相关的突变为K670N/M671L(Swedish)、1716V(Flo...5×FAD转基因小鼠(transgenic mice with five familial Alzheimer’s disease)是携带5个家族性基因突变的APP/PS1转基因小鼠,其中与β-淀粉样蛋白前体(amyloid precursor protein,APP)相关的突变为K670N/M671L(Swedish)、1716V(Florida)和V7171(London),与早老素-1(presenilin 1,PS1)相关的突变为MI46L和L286V。5×FAD小鼠在1.5月龄时脑内已有大量的β-淀粉样蛋白(β-amyloid,Aβ),2月龄时开始出现神经炎性斑(neuritic plaque,NP)。5×FAD小鼠的病理表型包括淀粉样斑块聚集、神经元丢失、神经胶质细胞增生和记忆功能障碍等。5×FAD小鼠的生物学特性可能涉及脑内Aβ斑块的形成变化、Tau蛋白过度磷酸化、突触功能障碍、神经炎症反应、线粒体功能障碍、血脑屏障损伤、神经元损伤、内质网应激和眼部病变等。作为阿尔茨海默病的经典动物模型,5×FAD转基因小鼠在早期即可模拟AD患者晚期的神经病理过程及行为学表现,被广泛应用于AD发病机制研究和AD新药开发。本文对5×FAD转基因小鼠模型的模型构建、生物学背景、生物学特性及AD防治药物的研发应用进行总结,以期为5×FAD转基因小鼠在AD研究中的应用提供参考与借鉴作用。展开更多
α-Linolenic acid(ALA,18:3Δ9,12,15)is an essential fatty acid for humans since it is the precursor for the biosynthesis of omega-3 long-chain polyunsaturated fatty acids(LC-PUFA).Modern people generally suffer from d...α-Linolenic acid(ALA,18:3Δ9,12,15)is an essential fatty acid for humans since it is the precursor for the biosynthesis of omega-3 long-chain polyunsaturated fatty acids(LC-PUFA).Modern people generally suffer from deficiency of ALA because most staple food oils are low or lack ALA content.Biotechnological enrichment of ALA in staple oil crops is a promising strategy.Chia(Salvia hispanica)has the highest ALA content in its seed oil among known oil crops.In this study,the FAD2 and FAD3 genes from chia were engineered into a staple oil crop,oilseed rape(Brassica napus),via Agrobaterium tumefaciens-mediated transformation of their LP4-2A fusion gene construct driven by the seed-specific promoter P_(NapA).In seeds of T0,T1,and T2 lines,the average ALA contents were 20.86,23.54,and 24.92%,respectively,which were 2.21,2.68,and 3.03 folds of the non-transformed controls(9.42,8.78,and 8.22%),respectively.The highest seed ALA levels of T0,T1,and T2 plants were 38.41,35.98,and 39.19%respectively,which were 4.10-4.77 folds of the respective controls.FA-pathway enzyme genes(BnACCD,BnFATA,BnSAD,BnSCD,BnDGAT1,BnDGAT2,and BnDGAT3)and positive regulatory genes(BnWRI1,BnLEC1,BnL1L,BnLEC2,BnABI3,BnbZIP67,and BnMYB96)were all significantly up-regulated.In contrast,BnTT1,BnTT2,BnTT8,BnTT16,BnTTG1,and BnTTG2,encoding negative oil accumulation regulators but positive secondary metabolism regulators,were all significantly down-regulated.This means the foreign ShFAD2-ShFAD3 fusion gene,directly and indirectly,remodeled both positive and negative loci of the whole FA-related network in transgenic B.napus seeds.展开更多
Hypersonic vehicles suffer from extreme aerodynamic heating during flights, especially around the area of leading edge due to its small curvature. Therefore, flush air data sensing(FADS) system has been developed to p...Hypersonic vehicles suffer from extreme aerodynamic heating during flights, especially around the area of leading edge due to its small curvature. Therefore, flush air data sensing(FADS) system has been developed to perform accurate measurement of the air data parameters. In the present study, the method to develop the FADS algorithms with fail-operational capability for a sharp-nosed hypersonic vehicle is provided. To be specific, the FADS system implemented with 16 airframe-integrated pressure ports is used as a case study. Numerical simulations of different freestream conditions have been conducted to generate the database for the FADS targeting in 2 ≤ Ma ≤ 5 and 0 km ≤ H ≤ 30 km. Four groups of neural network algorithms have been developed based on four different pressure port configurations, and the accuracy has been validated by 280 groups of simulations. Particularly, the algorithms based on the 16-port configuration show an excellent ability to serve as the main solver of the FADS, where 99. 5% of the angle-of-attack estimations are within the error band ±0. 2°. The accuracy of the algorithms is discussed in terms of port configuration. Furthermore, diagnosis of the system health is present in the paper. A fault-tolerant FADS system architecture has been designed, which is capable of continuously sensing the air data in the case that multi-port failure occurs, with a reduction in the system accuracy.展开更多
We have previously found that long-term effects of exposure to radiofrequency electromagnetic fields in 5xFAD mice with severe late-stage Alzheimer’s disease reduced both amyloid-βdeposition and glial activation,inc...We have previously found that long-term effects of exposure to radiofrequency electromagnetic fields in 5xFAD mice with severe late-stage Alzheimer’s disease reduced both amyloid-βdeposition and glial activation,including microglia.To examine whether this therapeutic effect is due to the regulation of activated microglia,we analyzed mic roglial gene expression profiles and the existence of microglia in the brain in this study.5xFAD mice at the age of 1.5 months were assigned to sham-and radiofrequency electromagnetic fields-exposed groups and then animals were exposed to 1950 MHz radiofrequency electromagnetic fields at a specific absorption rate of 5 W/kg for 2 hours/day and 5 days/week for 6 months.We conducted behavioral tests including the object recognition and Y-maze tests and molecular and histopathological analysis of amyloid precursor protein/a myloid-beta metabolism in brain tissue.We confirmed that radiofrequency electromagnetic field exposure for 6 months ameliorated cognitive impairment and amyloid-βdeposition.The expression levels of Iba1(pan-microglial marker)and colony-stimulating factor 1 receptor(CSF1R;regulates microglial prolife ration)in the hippocampus in 5xFAD mice treated with radiofrequency electromagnetic fields were significantly reduced compared with those of the sham-exposed group.Subsequently,we analyzed the expression levels of genes related to mic rogliosis and microglial function in the radiofrequency electromagnetic fields-exposed group compared to those of a CSF1R inhibitor(PLX3397)-treated group.Both radiofrequency electromagnetic fields and PLX3397 suppressed the levels of genes related to microgliosis(Csf1r,CD68,and Ccl6)and pro-inflammatory cytokine interleukin-1β.N otably,the expression levels of genes related to mic roglial function,including Trem2,Fcgr1α,Ctss,and Spi1,were decreased after long-term radiofrequency electromagnetic field exposure,which was also observed in response to microglial suppression by PLX3397.These results showed that radiofrequency electromagnetic fields ameliorated amyloid-βpathology and cognitive impairment by suppressing amyloid-βdeposition-induced microgliosis and their key regulator,CSF1R.展开更多
基金The study was financially supported by Projects from Shaanxi Province(2021LLRH-07-03-01 and 2023-ZDLNY-07)Yangling Seed Industry Innovation(YLzy-yc2021-01).The funders had no role in study design,data collection and analysis,decision to publish,or preparation of the manuscript.
文摘Genetic manipulation(either restraint or enhancement)of the biosynthesis pathway ofα-linolenic acid(ALA)in seed oil is an important goal in Brassica napus breeding.B.napus is a tetraploid plant whose genome often har-bors four and six homologous copies,respectively,of the two fatty acid desaturases FAD2 and FAD3,which con-trol the last two steps of ALA biosynthesis during seed oil accumulation.In this study,we compared their promoters,coding sequences,and expression levels in three high-ALA inbred lines 2006L,R8Q10,and YH25005,a low-ALA line A28,a low-ALA/high-oleic-acid accession SW,and the wildtype ZS11.The expression levels of most FAD2 and FAD3 homologs in the three high-ALA accessions were higher than those in ZS11 and much higher than those in A28 and SW.The three high-ALA accessions shared similar sequences with the pro-moters and CDSs of BnFAD3.C4 and BnFAD3.A3.In A28 and SW,substitution of three amino acid residues in BnFAD2.A5 and BnFAD2.C5,an absence of BnFAD2.C1 locus,and a 549 bp long deletion on the BnFAD3.A3 promoter were detected.The profile of BnFAD2 mutation in the two low-ALA accessions A28 and SW is different from that reported in previous studies.The mutations in BnFAD3 in the high-ALA accessions are reported for thefirst time.In identifying the sites of these mutations,we provide detailed information to aid the design of mole-cular markers for accelerated breeding schemes.
基金This work was supported by the National Natural Science Foundation of China(31871549,32001441 and 32272015)the Chongqing Research Program of Basic Research and Frontier Technology,China(cstc2015jcyjBX0143)+2 种基金the Fundamental Research Funds for the Central Universities,China(XDJK2020C038)the National Key R&D Program of China(2016YFD0100506)the Young Eagles Program of Chongqing Municipal Commission of Education,China(CY220219)。
文摘α-Linolenic acid(ALA,18:3Δ9,12,15)is an essential fatty acid for humans since it is the precursor for the biosynthesis of omega-3 long-chain polyunsaturated fatty acids(LC-PUFA).Modern people generally suffer from deficiency of ALA because most staple food oils are low or lack ALA content.Biotechnological enrichment of ALA in staple oil crops is a promising strategy.Chia(Salvia hispanica)has the highest ALA content in its seed oil among known oil crops.In this study,the FAD2 and FAD3 genes from chia were engineered into a staple oil crop,oilseed rape(Brassica napus),via Agrobaterium tumefaciens-mediated transformation of their LP4-2A fusion gene construct driven by the seed-specific promoter P_(NapA).In seeds of T0,T1,and T2 lines,the average ALA contents were 20.86,23.54,and 24.92%,respectively,which were 2.21,2.68,and 3.03 folds of the non-transformed controls(9.42,8.78,and 8.22%),respectively.The highest seed ALA levels of T0,T1,and T2 plants were 38.41,35.98,and 39.19%respectively,which were 4.10-4.77 folds of the respective controls.FA-pathway enzyme genes(BnACCD,BnFATA,BnSAD,BnSCD,BnDGAT1,BnDGAT2,and BnDGAT3)and positive regulatory genes(BnWRI1,BnLEC1,BnL1L,BnLEC2,BnABI3,BnbZIP67,and BnMYB96)were all significantly up-regulated.In contrast,BnTT1,BnTT2,BnTT8,BnTT16,BnTTG1,and BnTTG2,encoding negative oil accumulation regulators but positive secondary metabolism regulators,were all significantly down-regulated.This means the foreign ShFAD2-ShFAD3 fusion gene,directly and indirectly,remodeled both positive and negative loci of the whole FA-related network in transgenic B.napus seeds.
文摘Hypersonic vehicles suffer from extreme aerodynamic heating during flights, especially around the area of leading edge due to its small curvature. Therefore, flush air data sensing(FADS) system has been developed to perform accurate measurement of the air data parameters. In the present study, the method to develop the FADS algorithms with fail-operational capability for a sharp-nosed hypersonic vehicle is provided. To be specific, the FADS system implemented with 16 airframe-integrated pressure ports is used as a case study. Numerical simulations of different freestream conditions have been conducted to generate the database for the FADS targeting in 2 ≤ Ma ≤ 5 and 0 km ≤ H ≤ 30 km. Four groups of neural network algorithms have been developed based on four different pressure port configurations, and the accuracy has been validated by 280 groups of simulations. Particularly, the algorithms based on the 16-port configuration show an excellent ability to serve as the main solver of the FADS, where 99. 5% of the angle-of-attack estimations are within the error band ±0. 2°. The accuracy of the algorithms is discussed in terms of port configuration. Furthermore, diagnosis of the system health is present in the paper. A fault-tolerant FADS system architecture has been designed, which is capable of continuously sensing the air data in the case that multi-port failure occurs, with a reduction in the system accuracy.
基金Institute of Information&Communications Technology Planning&Evaluation(IITP)grant funded by Korea government(MSIT),Nos.2017-0-00961 and 2019-0-00102(to HDC)。
文摘We have previously found that long-term effects of exposure to radiofrequency electromagnetic fields in 5xFAD mice with severe late-stage Alzheimer’s disease reduced both amyloid-βdeposition and glial activation,including microglia.To examine whether this therapeutic effect is due to the regulation of activated microglia,we analyzed mic roglial gene expression profiles and the existence of microglia in the brain in this study.5xFAD mice at the age of 1.5 months were assigned to sham-and radiofrequency electromagnetic fields-exposed groups and then animals were exposed to 1950 MHz radiofrequency electromagnetic fields at a specific absorption rate of 5 W/kg for 2 hours/day and 5 days/week for 6 months.We conducted behavioral tests including the object recognition and Y-maze tests and molecular and histopathological analysis of amyloid precursor protein/a myloid-beta metabolism in brain tissue.We confirmed that radiofrequency electromagnetic field exposure for 6 months ameliorated cognitive impairment and amyloid-βdeposition.The expression levels of Iba1(pan-microglial marker)and colony-stimulating factor 1 receptor(CSF1R;regulates microglial prolife ration)in the hippocampus in 5xFAD mice treated with radiofrequency electromagnetic fields were significantly reduced compared with those of the sham-exposed group.Subsequently,we analyzed the expression levels of genes related to mic rogliosis and microglial function in the radiofrequency electromagnetic fields-exposed group compared to those of a CSF1R inhibitor(PLX3397)-treated group.Both radiofrequency electromagnetic fields and PLX3397 suppressed the levels of genes related to microgliosis(Csf1r,CD68,and Ccl6)and pro-inflammatory cytokine interleukin-1β.N otably,the expression levels of genes related to mic roglial function,including Trem2,Fcgr1α,Ctss,and Spi1,were decreased after long-term radiofrequency electromagnetic field exposure,which was also observed in response to microglial suppression by PLX3397.These results showed that radiofrequency electromagnetic fields ameliorated amyloid-βpathology and cognitive impairment by suppressing amyloid-βdeposition-induced microgliosis and their key regulator,CSF1R.
