An Effective Method for Raising Antisera Against β-defensins: Double-copy Protein Expression of mBin1b in E. coli

Defensins are a family of evolutionarily relatedvertebrate antimicrobial peptides with a characteristicβ-sheet-rich fold and a framework of six disulphide-linkedcysteines. The two main defensin subfamilies, α- andβ-defensin, differ in the length of pep…     

Sex differences in morphine-induced behavioral sensitization and social behaviors in ICR mice

Gender and genetic strain are two prominent variants that influence drug abuse. Although certain sexrelated behavioral responses have been previously characterized in ICR mice, little is known about the effects of sex on morphine-induced behavioral responses in this outbred strain. Therefore, in this study, we investigated the sex differences of morphine-induced locomotion, anxiety-like and social behaviors in ICR mice. After morphine or saline exposure for four consecutive days(twice daily), increased locomotion, more time spent in the central area, as well as attenuated rearing and self-grooming behaviors were found in morphine-treated females in an open field; no differences were found in locomotion and the time spent in the central area between male and female controls. When interacting with the samesex individuals, female controls were engaged in more social investigation, following, body contacting and self-grooming behaviors than controls; morphine exposure reduced contacting and self-grooming behaviors in females; in contrast, these effects were not found in males. These results indicate that female ICR mice are more prosocial and are more susceptible to morphine exposure than males.     

文昌鱼抗金葡菌感染差减文库的构建及免疫相关基因分析

适应性免疫的起源一直是免疫学研究的关键问题.文昌鱼被认为是最接近于脊椎动物的祖先 自从被发现以来一直是研究脊椎动物起源与进化机制的经典模式动物.为了在文昌鱼中寻找适应性免疫系统的分子证据,采用金黄色葡萄球菌感染文昌鱼以调查免疫的起源.应用抑制性差减杂交(SSH)技术,通过对差减文库克隆序列的测定,共获得588个表达序列标签(EST).对这些EST进行生物信息学分析和进一步功能分类,发现了一些免疫上调基因,如免疫调控基因、凋亡相关基因、细胞黏附相关基因、转录相关基因、信号传导相关基因等,以及一些非免疫相关基因;这些基因在文昌鱼中绝大多数为首次报道.金黄色葡萄球菌差减文库的成功构建,为调查文昌鱼抗细菌感染的分子事件提供了重要线索,对于这些新发现基因的进一步研究将有助于深入了解免疫系统起源与进化的机制.     

唇?硬骨蛋白基因克隆及其在肌间刺发生过程中的表达

为探究硬化蛋白(Sclerostin, SOST)与肌间刺发生之间的关系, 研究通过转录组测序和RT-PCR获得了唇?(Hemibarbus labeo)硬化蛋白基因cDNA序列。序列分析表明, 硬化蛋白由信号肽和成熟肽两部分组成, 成熟肽包含一个胱氨酸结样结构域。系统进化树分析表明, 唇?硬化蛋白与金鱼(Carassius auratus)硬化蛋白最为接近。通过荧光定量PCR检测发现唇?硬化蛋白基因mRNA在所有被检测的组织中均有表达, 在鳃中的表达量最高。RNA原位杂交结果表明, 硬化蛋白mRNA分布于肌膈, 且随着肌间刺的发生信号逐步减弱。RT-qPCR显示, 在肌间刺4个发育阶段中, 硬化蛋白基因mRNA的变化显著, 在阶段Ⅰ表达量最高, 随后逐步降低。综上, 唇?硬化蛋白与肌间刺骨化存在一定的相关性。研究结果将为进一步调查鱼类硬化蛋白功能及硬化蛋白在肌间刺形成过程中的作用提供理论依据。     

Estimating ancestral distributions of lineages with uncertain sister groups： a statistical approach to Dispersal-Vicariance Analysis and a case using Aesculus L. （Sapindaceae） including fossils

We propose a simple statistical approach for using Dispersal-Vicariance Analysis （DIVA） software to infer biogeographic histories without fully bifurcating trees. In this approach, ancestral ranges are first optimized for a sample of Bayesian trees. The probability P of an ancestral range r at a node is then calculated as P（rY） = ∑t^n=1 F（rY）t Pt where Y is a node, and F（rY） is the frequency of range r among all the optimal solutions resulting from DIVA optimization at node Y, t is one of n topologies optimized, and Pt is the probability of topology t. Node Y is a hypothesized ancestor shared by a specific crown lineage and the sister of that lineage ＂x＂, where x may vary due to phylogenetic uncertainty （polytomies and nodes with posterior probability 〈 100%）. Using this method, the ancestral distribution at Y can be estimated to provide inference of the geographic origins of the specific crown group of interest. This approach takes into account phylogenetic uncertainty as well as uncertainty from DIVA optimization. It is an extension of the previously described method called Bayes-DIVA, which pairs Bayesian phylogenetic analysis with biogeographic analysis using DIVA. Further, we show that the probability P of an ancestral range at Y calculated using this method does not equate to pp＊F（rY） on the Bayesian consensus tree when both variables are 〈 100%, where pp is the posterior probability and F（rY） is the frequency of range r for the node containing the specific crown group. We tested our DIVA-Bayes approach using Aesculus L., which has major lineages unresolved as a polytomy. We inferred the most probable geographic origins of the five traditional sections of Aesculus and ofAesculus californica Nutt. and examined range subdivisions at parental nodes of these lineages. Additionally, we used the DIVA-Bayes data from Aesculus to quantify the effects on biogeographic inference of including two wildcard fossil taxa in phylogenetic analysis. Our analysis resolved the geographic     

Differential responses to feeding by the tomato/potato psyllid between two tomato cultivars and their implications in establishment of injury levels and potential of damaged plant recovery

An invasive new biotype of the tomato/potato psyllid （Bactericera [Paratrioza] cockerelli [Sulc.]） （Homoptera： Psyllidae） recently has caused losses exceeding 50% on fresh market tomatoes in western North America. Despite these extensive losses, little is known regarding the threshold levels at which populations must be suppressed in order to prevent economic losses. A series of experiments were therefore designed using combinations of two common tomato cultivars （QualiT 21 and Yellow Pear）, five pest-densities （0, 20, 30, 40 and 50 nymphs/plant）, and three feeding-duration （5 days, 10 days, and lifetime） treatments to test the relative importance of pest density, feeding period, and cumulative psyllid-days to establish economic threshold levels for psyllids. The cultivars differed considerably in their response to the toxin injected by the psyllid nymphs. ‘Yellow Pear＇ plants could recover from feeding by up to 40 nymphs for as long as 10 d, whereas ‘QualiT 21＇ plants were irreparably damaged by densities of 20 nymphs feeding for only 5 days. On ‘Yellow Pear＇, all plant measurements such as the number of yellow leaves and plant height were significantly better correlated with cumulative psyUid-days than with either pest density or feeding duration. On ‘QualiT 21 ＇, all plant measurements other than the number of yellow leaflets and leaves were significantly better correlated with pest density than with feeding duration or cumulative psyUid-days, and pest density was a better predictor of psyUid damage. Potential reasons for the variable responses between cultivars and the implications for psyllid sampling and integrated pest management are discussed.     

Secretory/releasing proteome-based identification of plasma biomarkers in HBV-associated hepatocellular carcinoma

For successful therapy, hepatocellular carcinoma (HCC) must be detected at an early stage. Herein, we used a proteomic approach to analyze the secretory/releasing proteome of HCC tissues to identify plasma biomarkers. Serum-free conditioned media (CM) were collected from primary cultures of cancerous tissues and surrounding noncancerous tissues. Proteomic analysis of the CM proteins permitted the identification of 1365 proteins. The enriched molecular functions and biological processes of the CM proteins, such as hydrolase activity and catabolic processes, were consistent with the liver being the most important metabolic organ. Moreover, 19% of the proteins were characterized as extracellular or membrane-bound. For validation, secretory proteins involved in transforming growth factor-β signaling pathways were validated in plasma samples. Alphafetoprotein (AFP), metalloproteinase (MMP)1, osteopontin (OPN), and pregnancy-specific beta-1-glycoprotein (PSG)9 were significantly increased in HCC patients. The overall performance of MMP1 and OPN in the diagnosis of HCC remained greater than that of AFP. In addition, this study represents the first report of MMP1 as a biomarker with a higher sensitivity and specificity than AFP. Thus, this study provides a valuable resource of the HCC secretome with the potential to investigate serological biomarkers. MMP1 and OPN could be used as novel biomarkers for the early detection of HCC and to improve the sensitivity of biomarkers compared with AFP.     

Atomic force microscopy imaging of live mammalian cells

Atomic force microscopy (AFM) was used to examine the morphology of live mammalian adherent and suspended cells. Time-lapse AFM was used to record the locomotion dynamics of MCF-7 and Neuro-2a cells. When a MCF-7 cell retracted, many small sawtooth-like filopodia formed and reorganized, and the thickness of cellular lamellipodium increased as the retraction progressed. In elongated Neuro-2a cells, the cytoskeleton reorganized from an irregular to a parallel, linear morphology. Suspended mammalian cells were immobilized by method combining polydimethylsiloxane-fabricated wells with poly-L-lysine electrostatic adsorption. In this way, the morphology of a single live lymphoma cell was imaged by AFM. The experimental results can improve our understanding of cell locomotion and may lead to improved immobilization strategies.