The GNAQ T96S Mutation Affects Cell Signaling and Enhances the Oncogenic Properties of Hepatocellular Carcinoma

Hepatocellular carcinoma (HCC), the most common malignant tumor in the liver, grows and metastasizes rapidly. Despite advances in treatment modalities, the five-year survival rate of HCC remains less than 30%. We sought genetic mutations that may affect the oncogenic properties of HCC, using The Cancer Genome Atlas (TCGA) data analysis. We found that the GNAQ T96S mutation (threonine 96 to serine alteration of the Gαq protein) was present in 12 out of 373 HCC patients (3.2%). To examine the effect of the GNAQ T96S mutation on HCC, we transfected the SK-Hep-1 cell line with the wild-type or the mutant GNAQ T96S expression vector. Transfection with the wild-type GNAQ expression vector enhanced anchorage-independent growth, migration, and the MAPK pathways in the SK-Hep-1 cells compared to control vector transfection. Moreover, cell proliferation, anchorage-independent growth, migration, and the MAPK pathways were further enhanced in the SK-Hep-1 cells transfected with the GNAQ T96S expression vector compared to the wild-type GNAQ-transfected cells. In silico structural analysis shows that the substitution of the GNAQ amino acid threonine 96 with a serine may destabilize the interaction between the regulator of G protein signaling (RGS) protein and GNAQ. This may reduce the inhibitory effect of RGS on GNAQ signaling, enhancing the GNAQ signaling pathway. Single nucleotide polymorphism (SNP) genotyping analysis for Korean HCC patients shows that the GNAQ T96S mutation was found in only one of the 456 patients (0.22%). Our data suggest that the GNAQ T96S hotspot mutation may play an oncogenic role in HCC by potentiating the GNAQ signal transduction pathway.     

Cu-Fe-Cr原位复合材料的纤维相结构

通过感应熔炼和冷拔变形制备了Cu 16Fe 2Cr原位复合材料 ,将铜基体选择腐蚀后提取出了纤维 ,采用SEM和TEM观察分析了纤维相结构。在较低的应变量 (η =1.6 7)时 ,纤维不均匀 ,随着变形量的增大 (η =5 .42 ) ,纤维外形变得均匀。在 η =5 .42时 ,一个显著的特点是单根Fe Cr纤维分为一些由晶界隔开的平行亚单元 (宽度约为 10 0nm) ,通过亚单元共同的 [110 ]衍射获得了晶粒之间的相对取向 ,相邻晶粒的取向角在 3°～ 15°之间。     

Cu-Fe-Cr原位复合材料中的纤维相

通过感应熔炼、铸造、锻造和冷拔变形制备了Cu-16Fe-2Cr(质量分数,％)原位复合材料,最终线材(其轧制后的冷拔变形量达到η=ln(A_0/A)=5.42,A_0和A分别为冷拔起始和冷拔后线材的横截面面积)的抗拉强度为980 MPa。将Cu基体选择腐蚀后提取出纤维,采用SEM和TEM观察分析纤维相组织形态。在较低的应变量时,一些Fe—Cr纤维保持着与铸态树枝晶相同的bcc单晶结构,选区电子衍射表明纤维已经形成了〈110〉织构。在较高的应变量时,单根Fe-Cr纤维分为一些由晶界隔开的平行亚单元(宽度约为100 nm),通过亚单元共同的[110]衍射获得了晶粒之间的相对取向关系,相邻晶粒的偏差角在11°—82°之间。根据Hall-Petch关系讨论了原位复合材料的强度问题。     

幽门螺杆菌尿素酶B亚单位特异性抗体检测方法的建立与评价

目的建立并评价幽门螺杆菌(Hp)尿素酶B亚单位(UreB)特异性抗体的检测方法。方法应用纯化的重组HpUreB作为包被抗原,建立检测HpUreB特异性抗体的ELISA间接法。以Hp抗体Western blot检测作为“金标准”,评价所建立方法的真实性和可靠性。结果检测血清特异性IgG的灵敏度为100·0%,特异度为97·7%,阳性预测值为97·2%,阴性预测值为100·0%,约登指数为0·977,符合率为98·7%,试验的一致率为98·5%;检测血清中总的特异性Ig灵敏度为97·1%,特异度为95·3%,阳性预测值为94·4%,阴性预测值为97·6%,约登指数为0·925,符合率为96·2%,试验的一致率为97·8%;检测唾液中特异性sIgA的灵敏度为96·2%,特异度为94·5%,阳性预测值为89·3%,阴性预测值为98·1%,约登指数为0·907,符合率为95·1%,试验的一致率为97·5%;检测粪便标本中的特异性sIgA的灵敏度为92·0%,特异度为90·2%,阳性预测值为85·2%,阴性预测值为94·9%,约登指数为0·822,符合率为90·9%,试验的一致率为98·6%,CV值均小于15%。结论该检测方法真实性、可靠性、重复性良好,能满足临床标本检测的要求。     

狗心脏中I_(to)相关钾离子通道表达分析

目的探讨钾离子通道Kv4·3和Kv1·4在正常与心衰的心肌组织中的表达差别,为进一步阐明心衰的发病机理奠定基础。方法建立狗的心衰模型,用Western blot方法对正常与心衰的狗心肌组织中Kv4·3、Kv1·4及辅助亚基Mink的表达进行分析。结果在心衰时,Kv4·3表达有明显下降;Kv1·4表达有所上升,在心肌组织的不同部位,即外层和内层Kv1·4的表达明显不同;同时Mink的表达在心肌外层也有明显下降。结论心衰时,心肌组织中形成Ito的钾离子通道Kv4·3和Kv1·4以及辅助亚基Mink的蛋白表达变化之间可能存在着一些相关性。     

上贝氏体精细结构的研究

用透射电镜研究上贝氏体铁素体的形成。观察到贝氏体条由更小的薄片状亚单元组成。沿贝氏体铁素体条纵向靠近平直边的母相中存在间断的应变场，其间隔尺寸与薄片亚单元的厚度对应。由于亚单元的横向尺寸差构成边界轮廓台阶。相变弛豫使亚单元间界和应变场逐渐消失。     

Construction of Asymmetrical Hexameric Biomimetic Motors with Continuous Single‐Directional Motion by Sequential Coordination

The significance of bionanomotors in nanotechnology is analogous to mechanical motors in daily life. Here the principle and approach for designing and constructing biomimetic nanomotors with continuous single‐directional motion are reported. This bionanomotor is composed of a dodecameric protein channel, a six‐pRNA ring, and an ATPase hexamer. Based on recent elucidations of the one‐way revolving mechanisms of the phi29 double‐stranded DNA (dsDNA) motor, various RNA and protein elements are designed and tested by single‐molecule imaging and biochemical assays, with which the motor with active components has been constructed. The motor motion direction is controlled by three operation elements: (1) Asymmetrical ATPase with ATP‐interacting domains for alternative DNA binding/pushing regulated by an arginine finger in a sequential action manner. The arginine finger bridges two adjacent ATPase subunits into a non‐covalent dimer, resulting in an asymmetrical hexameric complex containing one dimer and four monomers. (2) The dsDNA translocation channel as a one‐way valve. (3) The hexameric pRNA ring geared with left‐/right‐handed loops. Assessments of these constructs reveal that one inactive subunit of pRNA/ATPase is sufficient to completely block motor function (defined as K = 1), implying that these components work sequentially based on the principle of binomial distribution and Yang Hui's triangle.     

钢中贝氏体相变热力学

应用综合理论分析的方法研究了钢中贝氏体相变热力学.指出以往的KRC,LFG模型不适于贝氏体相变驱动力的计算.在进行相变热力学分析的基础上,依据γ→αB γ1→BF γ1转变机制设计了新的计算模型,并估算了Bs温度下相变阻力为105 J/mol.指出相变不仅与驱动力有关,而且取决于原子扩散能力.在贝氏体上部温度区,可以依靠界面扩散进行台阶长大;在460 ℃以下的某一段温度,可能以原子热激活跃迁无扩散机制进行贝氏体铁素体形核-长大过程;在Ms点稍上一段温度,可能以切变方式进行.     

High-resolution subunit detection of glutamate receptor by ultrasmall gold nanoparticles

In this study, we aimed to increase the sensitivity of protein labeling using 1.4 nm gold nanoparticles and glutamate δ2 receptor (GluD2) from the postsynaptic membrane of the Purkinje cells. The very small marker size of the particles reduces the steric hindrance between antibodies leading to a higher labeling efficiency of more than one subunit per single receptor molecule. The nanoparticles are visible in 200 kV dark‐field scanning transmission electron microscope on freeze‐fractured carbon replica of nervous tissue after plasma cleaning treatment. The different elemental composition of nanoparticles as Au nanogold or CdS quantum dot can be distinguished by energy dispersive X‐ray spectroscopy. This method ensures detection of an average of three subunits per GluD2 and often labels all four of them with 1.4 nm Au nanoparticles. It is concluded that this high‐resolution microscopic method is useful for exploring the quaternary structure of membrane proteins. Microsc. Res. Tech. 75:1159–1164, 2012. © 2012 Wiley Periodicals, Inc.