非伤害性取样在无尾两栖类保护遗传学研究中的应用

为了改变传统的将两栖类动物处死后再从肌肉提取DNA的取样方法,采用非伤害性取样方法分别采集了虎纹蛙的血液和趾提取DNA,并与从其肌肉中提取的DNA进行了比较,结果显示:使用血液和趾提取的DNA产量与肌肉样本的DNA产量相当;随后,分别对3种方法取样的样本进行了线粒体16S rRNA和核DNA的微卫星PCR分析,结果表明:3种取样方法所获得的扩增效果相当,均能满足虎纹蛙的保护遗传学研究.因此认为,用非伤害性取样技术替代传统的肌肉取样的方法在无尾两栖类的保护遗传学研究中是切实可行的.

应用非伤害性取样提取番鸭毛囊组织总RNA

目的:寻求一种从番鸭毛囊组织中高效提取总RNA的方法。方法:探讨了伤害性取样(剪切皮肤毛囊)、非伤害性取样(直接拔取毛囊)2种不同毛囊取样法对总RNA提取质量的影响,并对常用RNA提取方法TRIzol法中研磨和组织匀浆步骤细节稍加改进,琼脂糖电泳检测总RNA质量。结果:2种毛囊取样方法均能提取出高质量的总RNA,其28S、18S和5S条带清晰可见,无DNA污染。结论:非伤害性取样法可作为番鸭毛囊组织总RNA提取的适用取样方法。

非伤害性取样方法获取的拟穴青蟹DNA质量研究

选取5只采自广西北海的野生拟穴青蟹（Scylla paramamosain）,分别采用非伤害性（血淋巴）和伤害性（肌肉组织）取样方法提取基因组DNA进行COI扩增和ISSR扩增,检测DNA质量。结果表明,非伤害性（血淋巴）和伤害性（肌肉组织）取样方法获得的拟穴青蟹全基因组DNA的OD260/OD280分别为1.75~2.01和1.81~1.94,全基因组DNA纯度均较高,质量不相上下。同时,两种取样方法获取的DNA扩增片段大小均在600bp左右,同一个体扩增带型也是一致的。非伤害性（血淋巴）取样方法提取的基因组DNA,在质量上可以媲美伤害性取样（肌肉组织）提取的DNA,可以用于其它分子生物学实验研究。

棘腹蛙非伤害性取样DNA提取及16S rRNA基因序列特征分析

为了改变将两栖类动物处死后从肌肉提取DNA的传统取样方法,试验采用非伤害性取样方法分别从棘腹蛙蹼和趾提取DNA,并以肌肉为对照进行PCR扩增比较。结果表明:蹼和趾的扩增效果与肌肉相当,均能满足获得高质量DNA的试验需求。试验测定了四面山和酉阳棘腹蛙群体的16SrRNA基因部分片段的序列,长度均为530 bp,膘嘌呤(A)、胸腺嘧啶(T)、鸟嘌呤(G)、胞嘧啶(C)的含量相近,分别为30.2%2、2.8%、21.5%、25.5%,30.4%、22.8%、21.5%、25.3%,2个序列间有6个变异位点,核苷酸差异率为1.13%,其中转换5个、颠换1个,转换与颠换比值为5.0。基于已知的棘腹蛙所有16S rRNA基因511 bp同源序列,获得的系统发生树的拓扑结构表明,各群体间遗传分化程度表现为与其分布的地理区域的临近关系及是否存在地理隔离相对应。

非伤害性触摸刺激对慢性坐骨神经痛大鼠脊髓背角c-fos与TrkA受体表达的影响

本研究拟观察非伤害性触摸刺激对慢性坐骨神经损伤(CCI)大鼠脊髓背角c-fos及TrkA受体表达的影响,探讨脊髓在慢性神经痛产生和维持中的作用.

珍稀濒危鸟类微量取样及DNA提取的研究

为解决珍稀濒危鸟类分子生物学研究中存在的取样局限性,采用非伤害性取样和非损伤性取样,对大鸨10～70μL的血液和羽毛的不同部位(羽根、羽髓、羽鞘、实心羽茎和羽毛上脐周围绒羽)的DNA提取进行了研究.结果表明,鸟类血液样本与哺乳动物样本极为不同,两者相同体积的血液中,鸟类样本DNA含量是哺乳动物样本的几百倍,研究发现,大鸨可以从微量的血液中(10μL)提取出较多的DNA(45.75μg).对于羽毛的不同部位,羽髓、羽鞘和羽根可以提出一定纯度的核DNA和mtDNA;实心羽茎可以提出一定纯度的mtD-NA.因此该研究将拓宽珍稀濒危鸟类采样的范围,为珍稀濒危鸟类的分子生物学研究提供了更广阔的样品来源.

不同部位取样对鸡基因组DNA提取的效果的影响

采用伤害性取样和非伤害性取样,采集鸡的肝脏、血液、羽髓、羽鞘、羽根、实心羽茎和羽毛上脐周围绒羽等不同部位样品,并采取对样品不同处理程序提取基因组DNA。结果表明,鸡的微量血液(50μl)和羽髓样品提取基因组DNA与肝脏(50 mg)样品提取的基因组DNA的量相当;羽鞘、羽根、实心羽茎和羽毛上脐周围绒羽可以提出一定量的基因组DNA,其提取量少于肝脏和羽髓组织,但可以满足各种涉及PCR反应的分子生物学试验,肝脏羽髓消化最适时间为2.5 h,其他样品需消化过夜。该研究结果为禽类分子生物学研究拓宽了样本来源。

Data-based model free hysteresis identification for a nonlinear structure

In the last two decades, the damage detection for civil engineering structures has been widely treated as a modal analysis problem and most of the currently available vibration-based system identification approaches are based on modal parameters, namely the natural frequencies, mode shapes and damping ratios, and/or their derivations, which are suitable for linear systems. Nonlinearity is generic in engineering structures. For example, the initiation and development of cracks in civil engineering structures as typical structural damages are nonlinear process. One of the major challenges in damage detection, early warning and damage prognosis is to obtain reasonably accurate identification of nonlinear performance such as hysteresis which is the direct indicator of damage initiation and development under dynamic excitations. In this study, a general data-based identification approach for hysteretic performance in form of nonlinear restoring force using structural dynamic responses and complete and incomplete excitation measurement time series was proposed and validated with a 4-story frame structure equipped with smart devices of magneto-theological （MR） damper to simulate nonlinear performance. Firstly, as an optimization method, the least-squares technique was employed to identify the system matrices of an equivalent linear system of the nonlinear structure model basing on the exci- tation force and the corresponding vibration measurements with impact test when complete and incomplete excitations; and secondly, the nonlinear restoring force of the structure was identified and compared with the test measurements fi- nally. Results show that the proposed data-based approach is capable of identifying the nonlinear behavior of engineering structures and can be employed to evaluate the damage initiation and development of different structure under dynamic loads.