淫羊藿苷调控mTOR/Akt/CREB通路对高糖诱导的足细胞自噬及凋亡的影响

目的 探讨淫羊藿苷对高糖诱导的足细胞自噬、凋亡及哺乳动物雷帕霉素靶蛋白(mTOR)/丝氨酸苏氨酸蛋白激酶(Akt)/环磷酸腺苷反应元件结合蛋白(CREB)通路的影响。方法 将小鼠足细胞MPC5分为5组:正常对照组(5.5 mmol·L^(-1)葡萄糖)、高糖组(30 mmol·L^(-1)葡萄糖)、淫羊藿苷组(30 mmol·L^(-1)葡萄糖+5μmol·L^(-1)淫羊藿苷)、GDC-0349组(30 mmol·L^(-1)葡萄糖+50μmol·L^(-1)GDC-0349)、淫羊藿苷+GDC-0349组(30 mmol·L^(-1)葡萄糖+5μmol·L^(-1)淫羊藿苷+50μmol·L^(-1)GDC-0349)。培养48 h后,噻唑蓝法检测MPC5细胞活力;吖啶橙染色观察MPC5细胞自噬情况;流式细胞术检测MPC5细胞凋亡;蛋白印迹法检测MPC5细胞自噬[微管相关蛋白1轻链3(LC3)Ⅱ、LC3Ⅰ、自噬相关蛋白(Beclin-1)]、凋亡[Bcl-2相关X蛋白(Bax)、B淋巴细胞瘤-2(Bcl-2)]和mTOR/Akt/CREB通路相关蛋白的表达。结果 与正常对照组比较,高糖组MPC5细胞活力、Bcl-2、磷酸化mTOR(p-mTOR)/mTOR、磷酸化Akt(p-Akt)/Akt、磷酸化CREB(p-CREB)/CREB蛋白表达水平显著降低(P<0.05),自噬能力增强,自噬体表现出橙色荧光,细胞凋亡率、LC3Ⅱ/LC3Ⅰ、Beclin-1、Bax蛋白表达水平显著升高(P<0.05)。与高糖组比较,淫羊藿苷组MPC5细胞活力、LC3Ⅱ/LC3Ⅰ、Beclin-1、Bcl-2、p-mTOR/mTOR、p-Akt/Akt、p-CREB/CREB蛋白表达水平显著升高,自噬能力进一步增强,自噬体数量增多,自噬体呈现出砖红色荧光(P<0.05),细胞凋亡率、Bax蛋白表达水平显著降低(P<0.05);GDC-0349组MPC5细胞活力、LC3Ⅱ/LC3Ⅰ、Beclin-1、Bcl-2、p-mTOR/mTOR、p-Akt/Akt、p-CREB/CREB蛋白表达水平显著降低,自噬能力减弱,自噬体数量减少,自噬体表现出橙色荧光(P<0.05),细胞凋亡率、Bax蛋白表达水平显著升高(P<0.05);淫羊藿苷+GDC-0349可逆转淫羊藿苷对高糖诱导MPC5细胞的作用效果(P<0.05)。结论 淫羊藿苷通过激活mTOR/Akt/CREB通路促进高糖诱导的足细胞自噬抑制细胞凋亡。

二阶非线性抛物方程的B样条有限元法

首先,用二次B样条有限元法求解Fisher-Kolmogorov(FK)方程,证明半离散格式与全离散格式解的稳定性与收敛性;其次,用Crank-Nicolson方法离散时间变量,得到近似解的收敛阶为O((Δt)^(2)+h^(3));最后,用数值算例验证了理论分析结果及B样条有限元法的有效性.

Effect of Ta addition on solidification microstructure and element segregation of IN617B nickel-base superalloy

IN617B nickel-base superalloy is considered as a good candidate material in 700℃advanced ultrasupercritical coal-fired power plants.The effect of Ta addition on solidification microstructure and element segregation of IN617B alloy was investigated by OM,SEM,TEM,EDS,EPMA and thermodynamic calculation.The results showed that the solidification microstructure exhibited a dendritic segregation pattern with many primary carbides distributed in interdendritic regions,such as network M_(6)C,lath M_(23)C_(6) and granular Ti(C,N).The addition of Ta promoted the precipitation of Ta-rich MC significantly inhibiting the precipitation of M_(6)C and M_(23)C_(6),and reduced the segregation degree of Al,Mo and Ti alloying elements.The addition of Ta decreased the melting temperature of MC carbide,but did not impact the solidification path,that was,L→γmatrix→MC or Ti(C,N)→M_(6)C→M_(23)C_(6),where MC and Ti(C,N)tended to form symbiotic microstructure with M_(6)C.This study will provide theoretical basis and data support for the alloy optimization and casting structure control of IN617B nickel-based superalloy.

ANALYSIS OF BENDING, VIBRATION AND STABILITY FOR THIN PLATE ON ELASTIC FOUNDATION BY THE MULTIVARIABLE SPLINE ELEMENT METHOD

In this paper, the bicubic splines in product form are used to construct the multi-field functions for bending moments, twisting moment and transverse displacement of the plate on elastic foundation. The multivariable spline element equations are derived, based on the mixed variational principle. The analysis and calculations of bending, vibration and stability of the plates on elastic foundation are presented in the paper. Because the field functions of plate on elastic foundation are assumed independently, the precision of the field variables of bending moments and displacement is high.

Effect of rare earth elements on the microstructure and property for magnesium alloy AM60B

The effects of rare earth elements on the microstructure and properties ofmagnesium alloy AM60B alloy were studied. Different proportions of rare earth elements were added toAM60B and the tensile tests were carried out under different temperatures. The experimental resultsshow that at room temperature the tensile strength of AM60B can be improved with the addition ofrare earth elements. The ductility of which at room or elevated temperature (120 deg C) can also beimproved, and the ductility is to some extent in proportion with the amount of rare earth elements.The ductility at 120 deg C is better than that at room temperature. The microstructure graphsdemonstrate that appropriate amount of rare earth elements (0.1 percent-0.2 percent, mass fraction)can fine AM60B's grain and improve its ductility.

THE CONSTRUCTION OF WAVELET-BASED TRUNCATED CONICAL SHELL ELEMENT USING B-SPLINE WAVELET ON THE INTERVAL

Based on B-spline wavelet on the interval （BSWI）, two classes of truncated conical shell elements were constructed to solve axisymmetric problems, i.e. BSWI thin truncated conical shell element and BSWI moderately thick truncated conical shell element with independent slopedeformation interpolation. In the construction of wavelet-based element, instead of traditional polynomial interpolation, the scaling functions of BSWI were employed to form the shape functions through the constructed elemental transformation matrix, and then construct BSWI element via the variational principle. Unlike the process of direct wavelets adding in the wavelet Galerkin method, the elemental displacement field represented by the coefficients of wavelets expansion was transformed into edges and internal modes via the constructed transformation matrix. BSWI element combines the accuracy of B-spline function approximation and various wavelet-based elements for structural analysis. Some static and dynamic numerical examples of conical shells were studied to demonstrate the present element with higher efficiency and precision than the traditional element.

Two 8-node quadrilateral spline elements by B-net method

Isoparametric quadrilateral elements are widely used in the finite element method, but the accuracy of the isoparametric quadrilateral elements will drop obviously deteriorate due to mesh distortions. Spline functions have some properties of simplicity and conformality. Two 8-node quadrilateral elements have been developed using the trian- gular area coordinates and the B-net method, which can ex- actly model the quadratic field for both convex and concave quadrangles. Some appropriate examples are employed to evaluate the performance of the proposed elements. The nu- merical results show that the two spline elements can obtain solutions which are highly accurate and insensitive to mesh distortions.

Development of quadrilateral spline thin plate elements using the B-net method

The quadrilateral discrete Kirchhoff thin plate bending element DKQ is based on the isoparametric element Q8, however, the accuracy of the isoparametric quadrilateral elements will drop significantly due to mesh distortions. In a previous work, we constructed an 8-node quadrilateral spline element L8 using the triangular area coordinates and the B- net method, which can be insensitive to mesh distortions and possess the second order completeness in the Cartesian co- ordinates. In this paper, a thin plate spline element is devel- oped based on the spline element L8 and the refined tech- nique. Numerical examples show that the present element indeed possesses higher accuracy than the DKQ element for distorted meshes.

Cluster and Condensed Phase Properties of Light Elements (especially B, C, N, Na and Si)

Amorphous forms of C will first be compared and contrasted with amorphous Si, the differences in chemical bonding being emphasized in relation to atomic structure. After a brief discussion of ordered layers of graphite and of BN, some attention will be focused on BN cages and polymers and on C nanotubes. Finally properties of Na metal clusters will be discussed, including fission of such charged clusters

Identification of a NF-кB site in the negative regulatory element (ε-NRAII) of human ε-globin gene and its binding protein NF-кB p50 in the nuclei of K562 cells

The developmental control of the human e-globin gene expression is mediated by transcription regulatory elements in the 5’ flanking DNA of this gene. Sequence analysis has revealed a DNA motif (GGGGAATTTGCT) similar to NF-кB consensus sequence resides in the negative regulatory element (-3028bp～ -2902bp, termed ε-NRAII) 5’ to the cap site of this gene. NRF DNA fragment (-3010bp～ -2986bp) containing the NF-кB motif similar sequence was synthesized and used in electrophoresis mobility shift assay (EMSA) and competitive analysis. Data showed that a protein factor from nuclear extracts of K562 cells specifically interacted with NRF DNA fragment. The synthetic NF DNA fragment (containing NF-кB consensus sequence) could competed for the protein binding, but MNF DNA fragment (mutated NF-кB motif) could not, suggesting that the binding protein is a member of NF-кB/Rel family. Western blot assay demonstrated that the molecular weight of NF-кB protein in the nuclei of K562 cells is 50ku. We suggested that NF-кB p50 may play an important role in the regulation of human c-globin gene expression.

Identification of a NF-кB site in the negative regulatory element (ε-NRAII) of human ε-globin gene and its binding protein NF-кB p50 in the nuclei of K562 cells

The developmental control of the human e-globin gene expression is mediated by transcription regulatory elements in the 5' flanking DNA of this gene. Sequence analysis has revealed a DNA motif (GGGGAATTTGCT) similar to NF-кB consensus sequence resides in the negative regulatory element (-3028bp～ -2902bp, termed ε-NRAII) 5' to the cap site of this gene. NRF DNA fragment (-3010bp～ -2986bp) containing the NF-кB motif similar sequence was synthesized and used in electrophoresis mobility shift assay (EMSA) and competitive analysis. Data showed that a protein factor from nuclear extracts of K562 cells specifically interacted with NRF DNA fragment. The synthetic NF DNA fragment (containing NF-кB consensus sequence) could competed for the protein binding, but MNF DNA fragment (mutated NF-кB motif) could not, suggesting that the binding protein is a member of NF-кB/Rel family. Western blot assay demonstrated that the molecular weight of NF-кB protein in the nuclei of K562 cells is 50ku. We suggested that NF-кB p50 may play an important role in the regulation of human c-globin gene expression.

基于BDNF/TrkB/CREB通路研究六味地黄丸对丙戊酸钠诱导的孤独症谱系障碍模型仔鼠的作用机制

目的基于脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)/酪氨酸激酶受体B(tyrosine kinase receptor B,TrkB)/cAMP反应元件结合蛋白(cAMP response element binding protein,CREB)通路,探讨六味地黄丸对丙戊酸钠(sodium valproate,VPA)诱导的孤独症谱系障碍(autism spectrum disorder,ASD)仔鼠的作用机制。方法将13只SD孕鼠随机分为两组,其中10只孕鼠在第12.5天时腹腔注射VPA溶液(600 mg·kg^(-1))为VPA组,另外3只孕鼠注射等体积生理盐水为对照组。第21天对两组雄性仔鼠开展行为学检测,筛选出符合ASD疾病模型的仔鼠30只,随机分为模型组(等体积生理盐水),维生素D组(1480 IU·kg^(-1)),六味地黄丸高(3 g·kg^(-1))、中(1.5 g·kg^(-1))、低(0.75 g·kg^(-1))剂量组,每组6只。正常雄性仔鼠6只,设为空白组(等体积生理盐水)。各组仔鼠连续灌胃14 d,1次/d,给药后再次开展行为学检测。尼氏染色观察各组仔鼠海马组织神经元形态学变化,比色法检测各组仔鼠海马组织中谷氨酸(glutamic acid,GLU)、γ-氨基丁酸(gamma-aminobutyric acid,GABA)含量;qRT-PCR检测各组仔鼠海马组织中BDNF、TrkB、CREB mRNA相对表达。结果与对照组比较,VPA组仔鼠体质量、身长、尾长更小(P<0.05)。与空白组比较,模型组社交障碍症状明显(P<0.01),焦虑障碍症状明显(P<0.01),重复刻板行为增多(P<0.05或P<0.01),海马神经元结构损伤,GLU升高(P<0.01)、GABA下降(P<0.01),BDNF、TrkB、CREB mRNA表达降低(P<0.05或P<0.01);与模型组比较,维生素D组及六味地黄丸中、低剂量组仔鼠社交能力增强(P<0.05或P<0.01),焦虑障碍减轻(P<0.05或P<0.01),重复刻板行为减少(P<0.01或P<0.05),海马神经元结构明显复原,GLU下降(P<0.01),BDNF、TrkB、CREB mRNA表达增加(P<0.05或P<0.01),六味地黄丸中、低剂量组GABA上升(P<0.05或P<0.01)。结论六味地黄丸能显著改善VPA诱导的ASD仔鼠行为表现,增强海马组织神经元的再生与修复,其机制可能与平衡GLU、GABA水平,上调仔鼠海马组织中BDNF/TrkB/CREB的表达有关。

A cubic quadrilateral spline element with concave shapes

Basic requirement for applying isoparametric element is that the element has to be convex and no violent distortion is allowed. In this paper, a cubic quadrilateral spline element with 12 nodes has been developed using the triangular area coordinates and the B-net method, which can exactly model the cubic field for quadrilateral element with both convex and concave shapes. Neither mapping nor coordinate transformation is required and the spline element can obtain high accuracy solutions and insensitive to mesh distortions.

AB020.Inhibition of cyclic-AMP-response element binding protein and its impact on corneal wound healing in vitro and in vivo

Background:The cornea composes the outer surface of the eye and its transparency is required to allow light transmission to the retina.However,because of its position,the cornea is subjected to chemical and mechanical injuries that may lead to blindness.Our studies conducted using the human tissue-engineered cornea(hTEC)as a model provided evidence that the cyclic-AMP-response element binding protein(CREB)pathway is repressed during closure of corneal wounds.Based on these results,we hypothesized that closure of corneal wounds can be enhanced by preventing activation of CREB with the pharmacological inhibitor C646.Our goals were to proceed to the pharmacological inhibition of CREB(I)in vitro using the hTECs as a model,and then(II)in vivo using the rabbit as a model.Methods:The self-assembly approach was used to create hTECs,that were then wounded with an 8-mm diameter biopsy punch to create an epithelial defect.The tissues were then incubated with 10μM of C646(n=8).DMSO was used alone as a negative control(n=4).Closure of the wounds was monitored over a period of 5 days.Besides,the cornea of New Zealand white rabbits was debrided with an ethanol 70%solution to create an epithelial defect of 8-mm diameter.Several concentrations of C646(1,10,100μM et 1 mM)were applied as eye drops 3 times a day for up to 7 days.The wounded corneas(n=4 per concentration)were stained with fluorescein and photographed every day.Results:In vitro pharmacological inhibition of CREB with C646 considerably accelerated wound closure of all treated hTECs(4 days)compared to the control group(7 days).Moreover,the in vivo C646 treatment also accelerated wound healing of the corneas compared to the control group.The most effective concentration of C646 tested was the lowest(1μM),as it considerably enhanced the wound healing process.Conclusions:This study demonstrates that wound healing both in vitro and in vivo can be enhanced by preventing activation of CREB using a pharmacological inhibition approach.Most of all,this experiment suggests mediators from the CREB pathway as potential therapeutic targets on which we may influence to alter the wound healing dynamic of the cornea.We believe this study will lead to significant advancements in the clinical field of corneal defects.

MODEL B-I FOR MACRO RECTANGULAR ELEMENT AND PROBLEMS OF INFLUENCE SURFACE FOR PLATE STRUCTURES

Model B-I for marco rectangular element is presented for the first time in this paper. To establish the influence surf ace for resultant R of bending plates, a number of generalized distributive loads q are defined. It is shown by numerical examples that Model B-I and the formula for the generalized distributive loads advanced in this paper are featured by high accuracy, low memory space and flexibility in practical application, and that they are especially effective for plate structures subject to moving loads, such as the two-dimensional continuous plates of highway bridges and the flat stabs in piled jetty engineering.

Significance and identification of p53 response element binding sequence in the genome of hepatitis B virus

To explore the mechanism of interaction of hepatitis B virus (HBV) with P53 protein and its role in the hepatocarcinogenesis. Methods: HBV genome was analysed by computer program. The probe containing specific DNA-protein binding site in HBV genome was synthesized and reacted with nuclei protein of hepatoma cell lines. Specific binding was determined by electrophoretic mobility shift assay, electrophoretic mobility supershift assay and in situ ultraviolet cross-linking assay. Co-transfection was performed by using reporter gene cat, p53 and dexamethasone inducible HBx to observe the biological functions of HBV binding to p53 protein. Results: A p53 response element binding sequence is present in HBV genome at upstream of enhancer I from 1047 to 1059 hp. This sequence is capable of binding to p53 protein and increasing accumulation of p53 protein in cells. Conclusion: The results strongly suggest that DNA-protein binding of HBV with P53 protein plays a significant role and it may be the predominant mechanism in the pathogenesis of HBV-associated hepatocellular carcinoma.

电磁场B样条间断有限元方法

在计算电磁学领域,时域间断有限元算法(DGTD)一般基于模型空间的不规则网格划分和单元上高阶多项式插值计算。同样的插值阶数,二维空间四边形网格划分比三角形网格划分具有更少的自由度和更高的计算效率。然而,传统基于等参变换和多项式张量积插值的基函数空间在四边形单元上仅具有低阶完备性,且稳定性和精度受网格畸变影响较大。为此,提出了一种基于不规则四边形网格的高阶B样条插值DGTD方法,用于Maxwell方程的求解。文章采用的B样条基不仅具有高阶多项式空间的插值完备性,而且完全消除了单元内部自由度。此外,Maxwell方程离散系统的各系数矩阵还具有精确的解析形式。使用该方法分析腔体的本征模和楔形体的电磁散射,结果表明,相较于COMSOL软件最大允许时间步长提高2.5倍,计算所需未知量减少25%,证实了本文算法的高稳定性和高精度特点。

B、Zn元素掺杂力学增强型45S5生物活性玻璃支架

对45S5生物活性玻璃多孔支架进行B、Zn元素的掺杂,B元素有效降低了熔融法制备生物活性玻璃的熔融温度,减轻了高温对45S5玻璃生物活性的损害,SEM观察到掺B后支架显微结构变得更加致密,抗压测试结果显示掺B的玻璃支架抗压强度是纯45S5玻璃支架的2~3倍,可达9.68 MPa,表明了B对支架的力学性能亦有明显的提升作用。然而,掺入B元素会降低45S5玻璃支架的体外生物活性,过量的B掺杂还会导致支架被烧毁。在确定B元素的相对最佳掺杂浓度后,向玻璃支架中继续掺入Zn元素,能够提升支架的烧结稳定性,支架的抗压强度可继续增强,最优可达14.52 MPa,能与人体松质骨性能相匹配。同时,在SBF模拟体液中浸泡7天后支架的扫描电镜图展示了Zn元素的掺入,使得45S5玻璃支架的体外生物活性显著提升。总之,对45S5生物玻璃多孔支架进行B、Zn元素共掺,制备出了一种具有良好生物活性的力学增强型骨修复材料,具有一定骨组织工程运用价值。

高Ce含量RE-Fe-B烧结磁体元素分布与磁性能研究

为了开发具有商业价值的高Ce含量RE-Fe-B磁体,提高高丰度稀土Ce的利用率,对不同Ce替代量RE-Fe-B磁体的磁性能和微观结构进行了研究。实验结果表明,Ce/(Ce+Nd)质量分数达到40%的双主相磁体仍保持着优良的磁性能,剩磁Br达到1.31 T,最大磁能积(BH)_(max)达到310.56 kJ/m^(3)。在Ce替代量为40%的单、双主相磁体中,Nd和Ce元素的置换行为存在差异,单、双主相磁体中Nd元素均偏向于进入主相晶粒,Ce元素则趋于聚集在晶界相或主相晶粒表层,且在双主相磁体中形成贫Ce主相和接近名义成分的(Nd_(0.6)Ce_(0.4))_(30.5)(Fe,M)_(bal)B_(0.97)主相。通过观察磁畴翻转分析了单、双主相磁体的磁化行为,研究了双主相磁体矫顽力增强机制:贫Ce主相的高各向异性场对富Ce主相的磁矩起到了钉扎作用。Nd和Ce元素扩散行为的研究为制备高Ce含量、高磁性能的商业化RE-Fe-B磁体提供了依据。

某微型电动车侧碰条件下的B柱轻量化研究

B柱加强板作为B柱最主要的支撑结构,在侧碰中起到关键的作用,在满足侧碰安全性能的前提下进行轻量化研究具有重要意义。根据GB 20071—2006《汽车侧面碰撞的乘员保护》试验要求,利用HyperWorks建立有限元模型,基于Crachfem失效模式进行仿真分析,依托成形性分析,对焊缝位置进行优化,对比B柱轻量化前后侵入量和侵入速度的变化情况,并通过试验验证。结果表明:侧碰过程中,侵入量最大的位置位于B柱的R+400 mm的位置,此处为人体的胸部肋骨区域。通过激光拼焊的方式把B柱加强板的材料由DP780改为DP780+B1500HS,不仅侧碰安全性能得到提高,而且实现轻量化减重13.2%。