Safety of the express freight train running over a long-span bridge

Purpose-Express freight transportation is in rapid development currently.Owing to the higher speed of express freight train,the deformation of the bridge deck worsens the railway line condition under the action of wind and train moving load when the train runs over a long-span bridge.Besides,the blunt car body of vehicle has poor aerodynamic characteristics,bringing a greater challenge on the running stability in the crosswind.Design/methodology/approach-In this study,the aerodynamic force coefficients of express freight vehicles on the bridge are measured by scale model wind tunnel test.The dynamic model of the train-long-span steel truss bridge coupling system is established,and the dynamic response as well as the running safety of vehicle are evaluated.Findings-The results show that wind speed has a significant influence on running safety,which is mainly reflected in the over-limitation of wheel unloading rate.The wind speed limit decreases with train speed,and it reduces to 18.83 m/s when the train speed is 160 km/h.Originality/value-This study deepens the theoretical understanding of the interaction between vehicles and bridges and proposes new methods for analyzing similar engineering problems.It also provides a new theoretical basis for the safety assessment of express freight trains.

Research on Improvement Strategies of Psychological Training and Stage Expressiveness in Percussion Performance

With the continuous development of music education,percussion,as an important form of performance,has led to growing attention to the psychological training of its performers.This study aims to explore how psychological factors in percussion performance impact stage expressiveness and to propose corresponding psychological training strategies.By analyzing relevant domestic and international literature,we found that psychological training not only enhances performers’confidence and alleviates performance anxiety but also contributes to an overall improvement in performance quality.This study shows that methods such as emotional management and cognitive restructuring exhibit promising application potential in practice.Therefore,exploring a systematic psychological training program is significant for improving the stage expressiveness of percussion performers.

Effect of endurance training on the expression profile of circRNA-lncRNA-miRNA-mRNA in myocardial tissues of mice after exhaustive exercise

Objective:To clarify the effect of endurance training on the expression profile of circRNA-lncRNA-miRNA-mRNA in myocardial tissues of mice after exhaustive exercise.Methods:A total of 45 male C57BL/6 mice were randomly divided into control(C),low-strength endurance training(LSET)and high-strength endurance training(HSET)groups(n=15).The mice in the control group were not conducted to platform training.The mice in the LSET and HSET groups were conducted to platform training at 30%and 60%of exhaustive exercise once a day for 5 days a week,respectively.The exhaustion exercise was performed after 5 weeks of platform training.Total RNA was extracted from myocardial tissues,and the expression profile of circRNA-lncRNA-miRNA-mRNA in myocardial tissues was analyzed using Illimina transcriptome sequencing.Results:The distance and time of exhaustive exercise were longer in the LSET and HSET groups than in the control group,and the distance and time of exhaustive exercise were longer in the HSET group than in the LSET group(P<0.05).A total of 54 differentially expressed circRNAs(28 down-regulated and 26 up-regulated),7 differentially expressed lncRNAs(all down-regulated),3 differentially expressed miRNAs(1 down-regulated and 2 up-regulated)and 99 differentially expressed mRNAs(81 down-regulated and 18 up-regulated)were identified by transcriptome sequencing(P<0.05).Interaction network analysis revealed that ENSMUSG00000113041,MSTRG.79740,mmu-miR-374c-5p,18 down-regulated mRNAs and 3 up-regulated mRNAs formed a regulatory network.GO functional analysis revealed that the differentially expressed mRNAs were mainly enriched in primary metabolic processes and macromolecular synthesis and metabolic processes.KEGG pathway analysis revealed that the differentially expressed mRNAs were mainly enriched in complement and coagulation cascade pathways,estrogen signaling pathway and glucagon signaling pathway.Conclusion:Endurance training could alter the expression profile of circRNA-lncRNA-miRNA-mRNA in myocardial tissues of mice after exhaustive exercise,and these differentially expressed RNAs form a regulatory network that affects cardiomyocyte synthesis and metabolism and thus participates in the regulation of myocardial injury.

Cloning,localization and expression analysis of two fw2.2-like genes in small-and large-fruited pear species

Fruit size is one of the most important agronomic characters,which is mainly determined by cell number and cell size.However,our current knowledge about pear is largely unknown.Through counting of pear mesocarp cells at different stages,we found that the cell number,rather than the cell size,is responsible for the differences between small- and large-fruited cultivars.Fruit weight-2.2（fw2.2） is an important quantitative trait locus（QTL） affecting fruit weight in tomato and functions as a negative regulator in carpel cell division.To get more insights into this QTL in pear fruit development,we isolated two putative homologous fw2.2 genes,which were designated as fw2.2-like（PbFWL） genes.PbFWLs encode Cys-rich proteins with the CCXXXXCPC motif and belong to the PLAC8 superfamily.In addition,results from the subcellular localization indicated that PbFWLs were localized in the plasma membrane.The expression profile of the PbFWL genes by qRT-PCR showed they expressed higher in small-sized fruit cultivar than that in large-sized fruit cultivar during the cell division period.In summary,our data suggest that these two PbFWLs might be negatively related to the cell division in pear fruit.

Study on Dominant Expression of Regional Culture of the Qinling Mountains in Local Residential Buildings

Regional culture of the Qinling Mountains shows distinct features since it was born in the local outstanding ecological environment, study on local architecture is significant for the dominant expression of regional culture, protection of local environment, and echoing with the theme of ecological civilization construction. This paper, on the basis of the mutual infl uence and evolution of regional culture and style of local residential buildings, explored the reasons for the weakening of local architectural style, and specified the signifi cance of promoting local style of the living environment. By studying the infl uence of local natural environment and humanistic environment on architectural style along the northern foot of the Qinling Mountains, the paper explored the expression of regional culture in residential buildings, with Xian Garden(Xi'an Yuanzi) as an example, and aimed at giving useful help to the dominant expression of regional culture in modern residential buildings.

CD93 and GIPC expression and localization during central nervous system inflammation

CD93 and GAIP-interacting protein, C termius （GIPC） have been shown to interactively alter phagocytic processes of immune cells. CD93 and GIPC expression and localization during cen-tral nervous system inflammation have not yet been reported. In this study, we established a rat model of brain inlfammation by lipopolysaccharide injection to the lateral ventricle. In the brain of rats with inlfammation, western blots showed increased CD93 expression that decreased over time. GIPC expression was unaltered. Immunohistochemistry demonstrated extensive distribution of CD93 expression mainly in cell membranes in the cerebral cortex. After lipopoly-saccharide stimulation, CD93 expression increased and then reduced, with distinct staining in the cytoplasm and nucleus. Double immunolfuorescence staining in cerebral cortex of normal rats showed that CD93 and GIPC widely expressed in resting microglia and neurons. CD93 was mainly expressed in microglial and neuronal cell membranes, while GIPC was expressed in both cell membrane and cytoplasm. In the cerebral cortex at 9 hours after model establishment, CD93-immunoreactive signal diminished in microglial membrane, with cytoplasmic transloca-tion and aggregation detected. GIPC localization was unaltered in neurons and microglia. These results are the ifrst to demonstrate CD93 participation in pathophysiological processes of central nervous system inlfammation.

Theory Analysis of the Handover Challenge in Express Train Access Networks （ETAN）

To handle the handover challenge in Express Train Access Networks(ETAN).mobility fading effects in high speed railway environments should be addressed first.Based on the investigation of fading effects in this paper,we obtain two theoretical bounds:HOTiming upper bound and HO-Margin lower bound,which are helpful guidelines to study the handover challenge today and in the future.Then,we apply them to analyze performance of conventional handover technologies and our proposal in ETAN.This follow-up theory analyses and simulation experiment results demonstrate that the proposed handover solution can minimize handover time up to 4ms(which is the fastest one so far),and reduce HO-Margin to 0.16 dB at a train speed of 350km/h.

Expression, purification, and subcellular localization of phospholipase C in Dunaliella salina

Plants possess effective mechanisms to respond quickly to the external environment. Rapid activation of phosphatidylinositol-specific phospholipase C (PLC) enzymes occurs after a stimulus. The PLC in Dunaliella salina plays important roles in growth and stress responses. However, the molecular basis of PLC action in D. salina remains little understood. To gain insight into the potential biological functions of this enzyme, we cloned a phospholipase C gene from D. salina in a previous study, named DsPLC (GenBank No. KF573428). Here, we present the prokaryotic expression, purification, and characterization of the DsPLC gene. The entire coding region of DsPLC was inserted into an expression vector pET32a, and the DsPLC gene was successfully expressed in Escherichia coli. The DsPLC protein was purified and identified using a polyclonal antibody and western blotting. Expressing DsPLC fused with a green fluorescent protein (GFP) in onion showed that DsPLC-GFP was localized to the intracellular membrane. Quantitative real-time PCR analysis revealed that the relative expression of the DsPLC gene was induced significantly by 3.0-mol/L NaCl at 4 h. Our results support the importance of PLC enzymes in plant defense signaling. This study provides a basis for further functional studies of the DsPLC gene and for additional analysis of the potential roles of PLC enzymes in response to abiotic stress.

Cloned s-Lap Gene Coding Area, Expression and Localization of s-Lap/GFP Fusion Protein in Mammal Cells

s-Lap is a new gene sequence from pig retinal pigment epithelial(RPE) cells, which was found and cloned in the early period of apoptosis of RPE cells damaged with visible light. We cloned the coding area sequence of the novel gene of s-Lap and constructed its recombinant eukaryotic plasmid pcDNA3.1-GFP/s-lap with the recombinant DNA technique. The expression and localization of s-lap/GFP fusion protein in CHO and B_~16 cell lines were studied with the instantaneously transfected pcDNA3.1-GFP/s-lap recombinant plasmid. ~s-Lap/GFP fusion protein can be expressed in CHO and B_~16 cells with a high rate expression in the nuclei.

Molecular Cloning,Expression Analysis and Localization of Exo70A1 Related to Self Incompatibility in Non-Heading Chinese Cabbage(Brassica campestris ssp.chinensis)

The exocyst is a conserved protein complex,and required for vesicles tethering,fusion and polarized exocytosis.Exo70A1,the exocyst subunit,is essential for assembly of the exocyst complex.To better understand potential roles of Exo70A1 in non-heading Chinese cabbage（Brassica campestris ssp.chinensis）,we obtained the full-length cDNA of Exo70A1 gene,which consisted of 1 917 bp and encoded a protein of 638 amino acids.BlastX showed BcExo70A1 shared 94.9% identity with Brassica oleracea var.acephala（AEI26267.1）,and clustered into a same group with other homologues in B.oleracea var.acephala and Brassica napus.Subcellular localization analysis showed BcExo70A1 was localized to punctate structures in cytosol of onion epithelial cells.Results showed that BcExo70A1 was widely presented in stamens,young stems,petals,unpollinated pistils,roots and leaves of self compatible and incompatible plants.The transcripts of BcExo70A1 in non- heading Chinese cabbage declined during initial 1.5 h after incompatible pollination,while an opposite trend was presented after compatible pollination.Our study reveals that BcExo70A1 could play essential roles in plant growth and development,and is related to the rejection of self pollen in non-heading Chinese cabbage.

Expression and subcellular localization of a novel gene <i>Bm-X</i>of the silkworm, <i>Bombyx mori</i>

According to the large-scale sequencing of cDNA library from silkworm pupae, the cDNA of a novel gene with blank research background was identified and temporarily named Bm-X. The length of this cDNA is 778 bp. We obtained its ORF for further study by bioinformatics analysis. It is 444 bp and encodes 147 amino acid residues, with a predicted molecular weight (MW) of 16.4 kD and an isoelectric point (pI) of 3.69. In this study, we successfully constructed a recombinant plasmid pET-28a(+)-Bm-X and expressed it in Escherichia coli. We used the fusion protein rBm-X which purified by Niaffinity chromatography to produce polyclonal antibodies against Bm-X for Western blot analysis. The analysis revealed that Bm-X was expressed in the larval midgut, the epidermis and the silk gland. In addition, the subcellular localization analysis of silkworm ovary epithelial cells (BmN cells) showed that Bm-X protein was located both in cytoplasm and nucleus, and the signal was stronger in cytoplasm than in nucleus. Our findings indicate that Bm-X gene is a novel species-specificity gene and its expression product can be detected in tissues of the fifth silkworm instar larvae and BmN cells.

Local Shear Stresses Elicit Different Mechanical Response and Gene Expression from Complex Stresses

It is well established that living cells and tissues respond to mechanical forces such as flow-related shear stresses in blood or interstitial space and complex tractional stresses at cell-matrix contacts and cell-cell contacts.However,how different modes of forces impact mechanical and biological responses is elusive.Here we describe a strategy of using the three-dimensional magnetic twisting cytometry(3D MTC)technology to apply forces in any desired directions to the same living cell.We reveal that for a fixed stress amplitude and frequency,a live cell exhibits mechanical anisotropy and responds to a local shear stress differently from responding to a local complex stress by stretching chromatin and upregulating gene transcription to different levels,extending our previous finding on force-induced direct gene activation.This finding highlights the importance of force modes in impacting cellular mechanical and biological responses in living cells and tissues and may have implications in tissue patterning and embryonic development.

Cloning, Expression Pattern Analysis and Subcellular Localization of Resveratrol Synthase Gene in Peanut (<i>Arachis hypogaea</i>L.)

Resveratrol synthase (RS) is a key enzyme that plays a critical role in the resveratrol synthesis pathway. In this study, six RS genes were isolated and characterized from peanut variety “Zhenzhu Hong” by silico cloning and RT-PCR. Bioinformatics analysis showed that deduced amino acid sequences of the six cloned RS genes were highly conserved with a similarity from 95% to 99% when compared to the RS genes which had been deposited at the GenBank. The results of amino acid sequences analysis showed six RS proteins contained the Chal_Sti_Synt_N and ACP_Syn_III_C domains and can be classified to same family but with different evolutionary distance. Expression pattern analysis by QRT-PCR provided evidence indicating that the mRNA of six RS genes were primarily expressed in the peanut shell at different developmental stages with different expression levels, but only lower levels of them were evident in the peanut kernel. The subcellular localization of RS protein in onion epidermal cell was performed by Agrobacterium tumefaciens-mediated transformation and the green fluorescent was monitored by confocal fluorescence microscopy. The results indicated that, RS1 and RS5 were located in the nucleus and plasma membrane respectively, while the RS2, RS3, RS4 and RS6 were located in both nucleus inner membrane and plasma membrane. The data will provide basic information for elucidating the regulatory mechanisms and enzyme kinetics underlying the RS genes in the resveratrol synthase pathway.

Intracellular Localization of ABC Transporter TAPL Differs between Transient and Stable Expression

Transporter associated with antigen processing (TAP)-like (TAPL;ABCB9) is a half-type ABC transporter with sequence similarity to TAP1 and TAP2 that function in the ER membrane. To determine the cellular localization, TAPL and truncated forms of it were tagged with GFP at their car-boxyl termini. Intracellular localization of these fusion proteins was compared between transient and stable expression in CHO-K1 cells. When they were expressed transiently, the fluorescence of the fusion proteins was detected on the intracellular membrane, mainly in the ER, and all the fusion proteins, i.e., TAPL(M1 -A766)-GFP, TAPL(M1-S275)-GFP, TAPL(M1-K182 )-GFP, TAPL(M 1-R141)-GFP and TAPL(M1-G75), were co-localized with an ER marker, PDI. However, the fluorescence of all of them except for TAPL(M1-G 75)-GFP and TAPL(M1-S275)-GFP overlapped with a lysosome marker, cathepsin D, upon stable expression. Lysosomal localization was similarly observed with TAPL(M1- A766)-DsRed, which was stably expressed. These results suggest that TAPL is sorted to the lysosomal membrane when expressed stably in CHO-K1 cells. Furthermore, the lysosomal targeting signal may comprise the N-terminal four transmembrane helices since the N-terminal two transmembrane helices may not be enough to function as such a signal.

Study on the Applied Talents Training in Local Colleges

It is the inevitable requirement to emphasize applied talents training in modem society when the economy and society are under rapid development, during which the local colleges are obliged to undertake this significant responsibility. Taking the training of applied talents as the core during the teaching and education is the important step taken by local colleges in Jilin to adapt to the social demands. The quality of the talents would directly influence the contributions made to the society by local colleges and its own sustainable development. Nowadays, the local colleges in Jilin should reinforce their efforts in the training of applied talents and deepen the reform comprehensively from the educational concepts, training programs and teaching courses so as to effectively realize the teaching objectives of the training on applied talents.

Reflections on Talents Training Mode of Music Education Major in Local Normal Universities

The local normal university is the cradle of training music teachers for rural primary and secondary schools. The talent training of the music education major in local normal universities has a direct influence on the basic music education reform in rural areas, and also decides the survival and development of the music education major in local normal universities. Based on the definition of the talent training mode of the music education major, this paper deeply majoring in music education in local normal universities. discusses the special quality and the training strategies of talents

Expression of transforming growth factor-β in local bony callus in traumatic brain injury combined with extremity fracture in rats

Objective To investigate gene expression of transforming growth factor-β(TGF-β)in local bony callus in tracumatic brain in jury combined with extremity long bone fracture in rats.Methods Eighty male SD rats were randomized into 2 even

小麦倒春寒响应基因TaJAZ6的克隆、表达模式及亚细胞定位分析

JAZ蛋白在植物生长发育和胁迫信号通路中具有关键作用。为探究JAZ蛋白在小麦倒春寒中的调控机制,从小麦幼穗中克隆了TaJAZ6基因,并对其分子特征、表达特性与亚细胞定位进行分析。结果表明,该基因CDS序列全长为549 bp,编码178个氨基酸,预测编码蛋白质的分子质量为18.376 ku,理论等电点为9.37,不稳定系数为62.44,属于不稳定蛋白。该基因编码蛋白质具有1个TIFY结构域和1个CCT_2结构域。系统进化树分析发现,该蛋白质与野生二粒小麦和乌拉尔图小麦TIFY 11b蛋白亲缘关系最近。TaJAZ6基因启动子区除含有CAAT-box、TATA-box等基础作用元件外,还含有激素类响应元件、光响应元件、低温响应元件、防御和应激反应元件等。实时荧光定量PCR分析发现,TaJAZ6基因在根、茎、叶和幼穗中均有表达,根系中表达量最高。TaJAZ6基因还受低温和茉莉酸甲酯(MeJA)的诱导表达,低温胁迫下,TaJAZ6在矮抗58(耐倒春寒)和郑麦366(倒春寒敏感)根、茎和叶中表达趋势相同,均显著升高;喷施300,350μmol/L的MeJA之后再低温处理TaJAZ6在2个小麦品种中表达量均显著下降。TaJAZ6在低温胁迫后的幼穗中表达量出现相反的趋势,在矮抗58幼穗中表达量显著下降,在郑麦366幼穗中则显著上升,推测该基因可能负调控了小麦倒春寒胁迫防御反应。通过喷施MeJA显著降低了低温胁迫下2个小麦品种幼穗中TaJAZ6基因的相对表达量,同时提高了小麦的结实粒数。亚细胞定位试验显示,TaJAZ6蛋白定位于细胞核中。以上研究结果表明,TaJAZ6可能在小麦响应倒春寒胁迫过程中发挥重要作用。

四季秋海棠BsMYB62的抗旱性功能研究

【目的】以前期从四季秋海棠叶片中克隆得到的R2R3型MYB转录因子BsMYB62为研究对象,分析其在干旱胁迫下的功能。【方法】对四季秋海棠MYB62蛋白进行亚细胞定位,并通过实时荧光定量PCR方法明确BsMYB62基因在四季秋海棠根、茎、叶、雄花和雌花5个组织部位中的表达情况;构建BsMYB62基因的过表达载体转化到拟南芥,观察过表达转基因株系与野生型植株在萌发期和幼苗期遭受干旱胁迫时的生长表型、种子萌发和根长情况,测定各株系在幼苗期干旱胁迫处理时的叶绿素荧光参数PSⅡ最大光化学效率(F_(v)/F_(m)),叶绿素、丙二醛(MDA)、脯氨酸(Pro)含量,抗氧化酶(SOD、POD、CAT)活性以及转基因株系中BsMYB62基因的相对表达量变化。【结果】BsMYB62定位于四季秋海棠的细胞核,其在根、茎、叶和雌雄花中均有表达,其中以根和茎中的表达水平较高。在MS培养基中,野生型拟南芥和3个BsMYB62过表达转基因株系(OE1、OE2和OE3)的发芽率和长势均无显著差异,而在200 mmol/L甘露醇(模拟干旱胁迫)培养基中,OE1、OE2和OE3在胁迫处理后前2 d的发芽率及主根长均显著高于野生型拟南芥,幼苗长势也明显优于野生型。置于培养土中干旱胁迫1 d时,OE1、OE2和OE3的BsMYB62即被显著诱导;干旱胁迫13 d时,OE1、OE2和OE3的F_(v)/F_(m)、叶绿素含量、Pro含量以及SOD、POD和CAT活性均显著高于野生型拟南芥,而MDA含量则显著低于野生型拟南芥。【结论】BsMYB62基因通过提高转基因拟南芥的光合潜能和抗氧化能力,显著增强其对干旱胁迫的抗性。

师范专业认证背景下地方综合性高校地理科学专业人才培养模式构建──以宁波大学为例

师范专业认证是衡量高校师范素质教育的一个重要指标,对指导地方综合性大学教师教育专业的人才培养具有重要意义。立足师范认证的核心理念,结合宁波大学地理科学(师范)的专业现状,提出“1234”新课程体系人才培养模式。并在此基础上,对通识课程、专业课程、教师教育课程、综合实践课程等教育平台的建设和改革进行了探讨:以区域综合为特点,打造通识课程平台;以学生兴趣为导向,搭造专业课程平台;以师范性为中心,打造教师教育课程中心;以实践教学资源为依托,搭建综合实践课程平台。通过构建一套完整的评价与持续改进体系,以期为师范认证背景下的专业建设和人才培养提供参考。