胚胎干细胞体外分化研究进展

胚胎干细胞是来源于胚胎的、内细胞团的、具有多向分化潜能的细胞。随着胚胎干细胞,特别是分化机制研究的不断深入,胚胎干细胞分化为各种类型细胞的研究也取得了较大进展,从而为细胞移植、细胞治疗和组织工程等提供了一个新的途径。主要综述了小鼠和人胚胎干细胞向内、中、外三个胚层细胞类型分化的技术,以期为胚胎干细胞定向诱导分化的研究提供一些参考。     

Raccoon dogs (Nyctereutes procyonoides) are the new natural definitive hosts of Metagonimus hakubaensis

The definitive hosts of Metagonimus hakubaensis are reported to be hamsters, rats, mice, dogs, cats, chickens, and quails in experimental infection and Japanese water shrews in natural infection. Here we report that raccoon dogs are new natural definitive hosts of M. hakubaensis, based on morphological and molecular analyses of Metagonimus flukes collected from the host species from Aomori Prefecture, Japan. Moreover, M. hakubaensis recovered from raccoon dogs showed higher fecundity than those recovered from Japanese water shrews. Therefore, raccoon dogs were considered as a more suitable natural definitive host of M. hakubaensis than Japanese water shrews.     

Epidemiological investigations of trichostrongylid infections in young cattle in different parts of Norway

The occurrence of trichostrongylid nematodes in young cattle was studied in relation to climate and geographical location, by examination of herbage and faecal samples from 8 farms in 7 different regions in Norway, during 3 consecutive summers. Species of Ostertagia and Cooperia were dominant. Infective larvae, especially those of Ostertagia, were able to survive the winter in the pasture to a great extent in all the areas studied. The spring larval counts varied considerably from place to place, and from year to year, but were often at fairly high levels. This confirms previous findings that the larvae which overwinter on the pasture may be a major cause of losses. Overt clinical disease, however, was only seen at 1 farm during this investigation.The large number of larvae often seen in the autumn seems to be of less significance as a cause of disease, due to the late appearance, but is of major importance as a source of the larvae which survive the winter in the pasture and infect young cattle the following spring.There was no obvious correlation between pasture larval counts and the climate, except that there seemed to be higher counts in the warmer parts of the country, where the grazing season is also longer. The only place were both faecal egg counts and larval counts in the herbage were consistently low, was in the very far north.     

SMAD2介导的Activin A信号对人胚胎干细胞向限定性内胚层细胞诱导分化的促进作用

【目的】研究SMAD2在Activin A促进人胚胎干细胞向限定性内胚层诱导分化中的作用。【方法】在人胚胎干细胞中转染SMAD2基因siRNA或者过表达质粒后,收集经Activin A分别诱导0,6,12,24,48,72,96和120 h的细胞各100 ng/mL,采用实时定量PCR检测SMAD2 与内中胚层共同前体标记Brachyury和内胚层标记Sox17的表达,进一步通过Western-blot分析Activin A诱导中SMAD2和磷酸化SMAD2（p-SMAD2）表达的变化。【结果】在Activin A诱导人胚胎干细胞向限定性内胚层细胞分化的过程中,干扰SMAD2 后48 h时才检测到Brachyury强表达,而单纯Activin A处理组24 h就检测到强表达;Sox17的表达始终较单纯Activin A处理组明显降低,因此,干扰SMAD2直接抑制了Activin A的诱导作用。而过表达SMAD2,Brachyury和Sox17的表达水平较单纯Activin A处理组明显增加,促进了限定性内胚层的发生;并且在Activin A诱导过程中,p-SMAD2的表达水平明显提高,而SMAD2的表达没有明显改变。【结论】SMAD2作为关键因子,介导了Activin A诱导人胚胎干细胞向限定性内胚层的分化,并转录调控Brachyury和Sox17的表达;SMAD2的磷酸化,激活并介导了Activin A诱导的信号转导通路。     

香根草分区定肥试验报告

为了构建节约型水保，探索种植香根草经济合理的施肥量，应用作物报酬递减律的原理，在莒口虎山开展了7个处理、3个重复和1个对照，共28个小区两种肥料（干猪粪和钙镁磷）的分区定肥试验研究。结果显示：施用猪粪增产效果明显，施用钙镁磷增产效果欠佳。结论：株施干猪粪11．6g增产效果最明显，其为当地适宜的经济合理施肥量。     

Promoting effects of activated Nodal signal pathway on definitive endoderm induction from mouse embryonic stem cells

AIM: To study the process of promoting mouse embryonic stem cells (ESC) to specify to definitive endoderm by up-regulating of Nodal signal pathway in order to find the best cultivated systems of differentiation of mouse ESC to definitive endoderm cells. METHODS: The cells were divided into different groups based on the culture medium: ESC group (serum-free medium + LIF), natural differentiation group (serum-free medium) and activin A group (serum-free medium +50μg/L activin A). The cells and the sterilized coverslips with cells were collected at 1, 3, 5 and 7 d of the cultivation. The proportion of CXCR4⁺ cells was detected by flow cytometry. The expression of CXCR4 was determined by immunocytochemical method, and the protein expression of OCT4 and CXCR4 was detected by Western blot. RESULTS: The proportion of CXCR4⁺ cells showed no dramatic change in ESC group along with the extending of cultivation day, while there were gradually increased in natural differentiation group and activin A group and the highest level was observed at 5 d. Among the 3 groups, the proportion of CXCR4⁺ cells at 5 d was the highest in activin A group. The brown or tan staining in the cells observed under microscope was considered as positive CXCR4 by immunocytochemistry. The protein levels of OCT4 and CXCR4 in ESC group along with the extending of cultivation days was observed. The expression levels of OCT4 were gradually decreased in the cells in natural differentiation group and activin A group, while those of CXCR4 were gradually increased with the highest level at 5 d. It was highest in the cells in activin A group. CONCLUSION: The proportion of definitive endoderm was the highest at 5 d of the induction during in vitro mouse ESC differentiation. Up-regulation of Nodal signal pathway by adding activin A at the early stage of mouse ESC differentiation promotes mouse ESC to specify to definitive endoderm with CXCR4 molecular marker.     

基于粪便样本的野生犬科动物物种鉴别与棘球绦虫感染调查

旨在探索粪便样本分析对犬科动物物种鉴别的方法及其可行性,同时对野外犬科动物棘球绦虫感染情况进行调查。采用哺乳动物线粒体DNA控制区通用引物对101份采集自野外环境的犬科动物粪便DNA进行PCR扩增和测序,通过核苷酸位点变异分析、BLAST相似性比对、遗传距离分析和系统进化树构建,对粪样来源物种进行分子鉴别。同时,采用粪-抗原ELISA检测,对粪样棘球绦虫感染情况进行调查。结果表明：粪样来源物种鉴别的检测成功率为73.27%,获得的74条序列共定义单倍型20个,所有单倍型序列均可在GenBank数据库中匹配到单一物种,序列相似度为98.52%~100%。单倍型根据序列差异可分为4个单倍型集,各单倍型集间平均碱基差异为17.83~70.10,各单倍型集内单倍型碱基差异在1~10个之间。各集合间遗传距离为0.068~0.342,远大于各集合内单倍型间遗传距离0.009~0.022。系统进化分析结果显示,同一集合的单倍型以高自展值聚集成一支。综合各项分析结果,可推测所检测粪样宿主来源分别为犬、灰狼、赤狐和红狐。粪-抗原ELISA检测发现阳性样本11份,样本棘球绦虫抗原阳性率为10.89%。综上表明：线粒体DNA控制区序列分析可用于犬科动物的分子鉴别,可结合核基因等其他分子标记进一步提升检测准确率。研究区域野外犬科动物棘球绦虫粪抗原阳性率较高。     

基于Excel规划求解的泵站加压灌溉管网优化设计研究

以管网系统年费用最小为目标函数,水泵扬程H和连续管径D为设计变量,建立了确定性的泵站加压灌溉管网优化设计模型,并应用Excel规划求解法对所建立的模型进行了求解。实例表明,在管网布置形式已知的情况下,与传统的经济流速法相比,Excel规划求解法是一种实用价值较高的模型求解方法,该方法不仅能够节省一定的工程投资,还提高了设计人员的设计水平和效率。     

Definitive Callose Deposition in Tapping Panel Dryness Affected Bark of Hevea brasiliensis

The structural and functional aspects of sieve elements in Tapping Panel Dryness (TPD) affected bark of Hevea brasiliensis, with special reference to definitive callose deposition, have been studied using histochemical methods. The effect of ethephon in definitive callose deposition in TPD affected bark also has been investigated, as the ethylene is reported to have a major role in TPD. The phloem of Hevea is characterized by sieve elements with compound sieve plates. The deposition of definitive callose on the sieve areas following TPD limits the translocation of sap through sieve elements. The amount of functional phloem was narrow and the accumulation of definitive callose was more in the sieve elements of TPD affected bark compared to that of unaffected bark of the same tree and of healthy trees. The affected area showed an increment in deposition of definitive callose in both vertical and horizontal extent of sieve areas; and there was a reduction in length of sieve plate, density of sieve pores, length of inter-sieve area wall, and number of sieve element rows in the inner bark. The lateral walls of non-functional sieve elements became thick and lignified. The anatomical changes in the sieve elements of TPD affected bark suggest that the translocation of phloem sap is regulated by deposition of definitive callose and less differentiation of sieve elements from vascular cambium. The external application of ethephon on bark enhanced the definitive callose deposition on sieve elements. The findings from the study reveal that ethylene plays a major role in the senescence of sieve elements.