三种固定液对两种氧化应激细胞模型Bclin1和LC3蛋白免疫荧光染色的影响

摘要 目的：比较采用三种不同的固定液对两种氧化应激细胞模型Beclin1和LC3蛋白免疫荧光染色的影响。方法：本研究使用丙酮/甲醇（1:1）固定液、甲醇固定液和4%多聚甲醛三种固定液分别对氧化应激细胞模型大鼠原代心肌成纤维细胞和MCF-7乳腺癌细胞株进行固定,然后再分别进行免疫荧光双染实验,对比三种固定液固定后对自噬关键调控蛋白Beclin1和LC3染色效果。结果：三种固定液对氧化应激细胞模型Beclin1和LC3蛋白免疫荧光染色结果存在较大差异。丙酮/甲醇（1:1）固定液固定后免疫荧光染色效果最佳,细胞结构清晰可见,两种蛋白定位表达清晰,甲醇固定液次之,4%多聚甲醛固定液效果欠佳。结论：在对大鼠原代心肌成纤维细胞和MCF-7乳腺癌细胞进行自噬相关蛋白免疫荧光双染色实验中,在使用其它固定液染色效果不佳的情况下,可以选择应用丙酮/甲醇（1:1）固定液固定,再进行免疫荧光染色;根据不同实验需求相应选择更适宜的固定液,以达到最佳的荧光染色结果。     

A comparison of microwave heating and proteolytic pretreatment antigen retrieval techniques in formalin fixed,paraffin embedded tissues

Antigen retrieval (AR) is a technique that re-exposes epitopes in formalin fixed, paraffin embedded sections and makes them detectable by immunohistochemistry. We compared the effects of two AR procedures, enzyme digestion and microwave heating, on immunostaining of vimentin and desmin in formalin fixed, paraffin embedded tissues. Our results showed that AR is necessary for vimentin and desmin immunostaining in tissues fixed in formalin for more than 48 h. With prolonged fixation times, microwave heating showed better results than enzyme digestion for AR. The same results were obtained using 1% zinc sulfate or Citra Plus solution as retrieval solutions for microwave heating. We recommend microwave heating for AR, because it is easier to use and produces better results compared to enzyme treatment.     

Acidiphilium cryptum DX1-1 CO2固定相关基因的克隆及在不同营养方式下的差异表达研究

目的：研究Acidiphilium cryptum DX1-1 CO2固定相关基因的克隆及在不同营养方式下的差异表达。方法：以Acidiphilium cryptum DX1-1（CCTCCM208056）的DNA为模板,基于A．cryptum JF-5同源功能基因序列（JGI,http：／／genome．oml．gov／cgi-bin／JGI_microbial／keggcategories．cgi）设计引物,对菌株DX1-1中的CO2固定相关基因Acry_0824,Acry_082,Acry_1067,Acry_1272,Acry_0022和Acry_0827进行了克隆和序列比对分析;并对它们在不同营养条件下的基因差异表达进行了分析。结果：从菌株DX1—1成功克隆了所选择的CO2固定相关基因,其序列与菌株JF的同源功能基因序列一致性分别达到了99．8％,99．6％,99．6％,99．5％,99．3％和99．8％;Acry_0824,Acry_1272和Acry_0827三个基因在各种混合养条件下表达均上调,说明它们在DX1—1 CO2固定中起较关键的作用。在加入0．1％的葡萄糖混合养条件下,DX1—1细胞明显利用空气中的CO2来生长和累积PHB。结论：限制性葡萄糖可以促进细胞自养生长和累积PHB。     

单端固定式下颌骨修复体的应力分析

目的:针对包括一侧髁状突的下颌骨缺损,通过有限元应力分析,了解单端固定式下颌骨修复体在功能运动时的受力与变形规律,以期寻求更加合理的修复体的设计和固定方式。方法:建立下颌骨断端和修复体的简易三维模型,模拟咀嚼运动,施加垂直方向载荷,进行有限元法应力分析,计算出该模型各组成部分的应力分布和受力变形。结果:在该模型加载时,延伸板基部和近断端处上部的螺钉颈部是应力集中的部位,近断端处下部的螺钉颈部和修复体的远端舌侧为形变最大的部位。结论:单端固定式下颌骨修复体在加载时,延伸板的基部和靠近断端的固定螺钉是应力集中的部位,修复体远离固定的一侧是变形最大的部位,提示我们应将延伸板形态设计为尽可能加宽,并应增加下颌骨下缘处的固定,使修复体与下颌骨断端受力更加合理,变形也尽可能缩小。     

Evolutionary dynamics of continuous strategy games on graphs and social networks under weak selection

Understanding the emergence of cooperation among selfish individuals has been a long-standing puzzle, which has been studied by a variety of game models. Most previous studies presumed that interactions between individuals are discrete, but it seems unrealistic in real systems. Recently, there are increasing interests in studying game models with a continuous strategy space. Existing research work on continuous strategy games mainly focuses on well-mixed populations. Especially, little theoretical work has been conducted on their evolutionary dynamics in a structured population. In the previous work (Zhong et al., BioSystems, 2012), we showed that under strong selection, continuous and discrete strategies have significantly different equilibrium and game dynamics in spatially structured populations. In this paper, we further study evolutionary dynamics of continuous strategy games under weak selection in structured populations. By using the fixation probability based stochastic dynamics, we derive exact conditions of natural selection favoring cooperation for the death–birth updating scheme. We also present a network gain decomposition of the game equilibrium, which might provide a new view of the network reciprocity in a quantitative way. Finally, we make a detailed comparison between games using discrete and continuous strategies. As compared to the former, we find that for the latter (i) the same selection conditions are derived for the general 2 × 2 game; especially, the rule b/c > k in a simplified Prisoner's Dilemma is valid as well; however, (ii) for a coordination game, interestingly, the risk-dominant strategy is disfavored. Numerical simulations have also been conducted to validate our results.     

Population genetic structure of the Plasmodium vivax circumsporozoite protein (Pvcsp) in Sri Lanka

Molecular methods elucidate evolutionary and ecological processes in parasites, where interaction between hosts and parasites enlighten the evolution of parasite lifestyles and host defenses. Population genetics of Plasmodium vivax parasites accurately describe transmission dynamics of the parasites and evaluation of malaria control measures. As a first generation vaccine candidate against malaria, the Circumsporozoite Protein (CSP) has demonstrated significant potential in P. falciparum. Extensive polymorphism hinders the development of a potent malaria vaccine. Hence, the genetic diversity of Pvcsp was investigated for the first time in 60 Sri Lankan clinical isolates by obtaining the nucleotide sequence of the central repeat (CR) domain and examining the polymorphism of the peptide repeat motifs (PRMs), the genetic diversity indices and phylogenetic relationships. PCR amplicons determined size polymorphism of 610, 700 and 710 bp in Pvcsp of Sri Lanka where all amino acid sequences obtained were of the VK210 variant, consisting variable repeats of 4 different PRMs. The two most abundant PRMs of the CR domain, GDRADGQPA and GDRAAGQPA consisted ~ 2-4 repeats, while GNRAAGQPA was unique to the island. Though, different nucleotide sequences termed repeat allotypes (RATs) were observed for each PRM, these were synonymous contributing to a less polymorphic CR domain. The genetic diversity of Pvcsp in Sri Lanka was due to the number of repetitive peptide repeat motifs, point mutations, and intragenic recombination. The 19 amino acid haplotypes defined were exclusive to Sri Lanka, whereas the 194 Pvcsp sequences of global isolates generated 57 more distinct a.a. haplotypes of the VK210 variant. Strikingly, the CR domain of both VK210 and VK247 variants was under purifying selection interpreting the scarcity of CSP non-synonymous polymorphisms. Insights to the distribution of RATs in the CR region with geographic clustering of the P. vivax VK210 variant were revealed. The cladogram reiterated this unique geographic clustering of local (VK210) and global isolates (VK210 and VK247), which was further validated by the elevated fixation index values of the VK210 variant.     

Discriminative whisking in the head-fixed rat: optoelectronic monitoring during tactile detection and discrimination tasks

We compared whisking movement patterns during acquisition of tactile detection and object discrimination under conditions in which (a) head movements are excluded and (b) exposure to tactile discriminanda is confined to the large, moveable vibrissae (macrovibrissae). We used optoelectronic instrumentation to track the movements of an individual whisker with high spatio-temporal resolution and a testing paradigm, which allowed us to dissociate performance on an “indicator” response (lever pressing) from the rat's “observing” responses (discriminative whisking). We analyzed the relation between discrimination performance and whisking movement patterns in order to clarify the process by which the indicator response comes under the stimulus control of information acquired by the rat's whisking behavior. Whisking patterns over the course of task acquisition differed with task demands. Acquisition of the Detection task was correlated with modulation of only one whisking movement parameter - total number of whisks emitted, and more whisking was seen on trials in which the discriminandum was absent. Discrimination between a sphere and cube differing in size and texture was correlated with a reduction in whisk duration and protraction amplitude and with a shift towards higher whisking frequencies. Our findings confirm previous reports that acquisition of tactile discriminations involves modulation by the animal of both the amount and the type of whisking. In contrast with a previous report (Brecht et al., 1997), they indicate that rats can solve tactile object detection and discrimination tasks (a) using only the large, motile mystacial vibrissae (macrovibrissae) and (b) without engaging in head movements.We conclude that the functional contribution of the macrovibrissae will vary with the nature of the task and the conditions of testing.     

The one-third law of evolutionary dynamics

Evolutionary game dynamics in finite populations provide a new framework for studying selection of traits with frequency-dependent fitness. Recently, a "one-third law" of evolutionary dynamics has been described, which states that strategy A fixates in a B-population with selective advantage if the fitness of A is greater than that of B when A has a frequency 13. This relationship holds for all evolutionary processes examined so far, from the Moran process to games on graphs. However, the origin of the "number"13 is not understood. In this paper we provide an intuitive explanation by studying the underlying stochastic processes. We find that in one invasion attempt, an individual interacts on average with B-players twice as often as with A-players, which yields the one-third law. We also show that the one-third law implies that the average Malthusian fitness of A is positive.     

Fixation in haploid populations exhibiting density dependence II: the quasi-neutral case

We determine fixation probabilities in a model of two competing types with density dependence. The model is defined as a two-dimensional birth-and-death process with density-independent death rates, and birth rates that are a linearly decreasing function of total population density. We treat the 'quasi-neutral case' where both types have the same equilibrium population densities. This condition results in birth rates that are proportional to death rates. This can be viewed as a life history trade-off. The deterministic dynamics possesses a stable manifold of mixtures of the two types. We show that the fixation probability is asymptotically equal to the fixation probability at the point where the deterministic flow intersects this manifold. The deterministic dynamics predicts an increase in the proportion of the type with higher birth rate in growing populations (and a decrease in shrinking populations). Growing (shrinking) populations therefore intersect the manifold at a higher (lower) than initial proportion of this type. On the center manifold, the fixation probability is a quadratic function of initial proportion, with a disadvantage to the type with higher birth rate. This disadvantage arises from the larger fluctuations in population density for this type. These results are asymptotically exact and have relevance for allele fixation, models of species abundance, and epidemiological models.     

The analysis of bi-level evolutionary graphs

Evolutionary graphs (EGs), in which evolutionary dynamic is arranged on a graph, were initially proposed by Lieberman et al. [Lieberman, E., Hauert, C., Nowak, M.A., 2005. Evolutionary dynamics on graphs. Nature 433, 312–316] in the biological field and many biological phenomena are successfully explained. EGs on two levels (or bi-level EGs), based on some biological phenomena, are considered in this paper. The bi-level EGs are compared with the one-rooted EGs in two cases. One has the identical numbers of the followers, the other with the same numbers of total individuals. Then, some properties of the bi-level EGs are obtained. It is showed that bi-level EGs are more stable, and the bi-level EGs with just two leaders are the most stable, if they have identical followers respectively. The bi-level EGs theory can successfully explain the phenomena of symbiosis in biology.