组织微阵列的原理及应用

组织微阵列是将若干石蜡包埋块上的各种典型组织或细胞转移到一个新石蜡块上重新构建微型化高通量组织阵列的方法,可以有效地进行各类临床病理组织的原位观察和研究。组织微阵列技术可以在一块薄片上同时排列几百个样品,进行高通量的基因和蛋白质分析,且能够充分利用组织资源,将多个样品的平行实验一次完成。在分析RNA和某些蛋白质时,石蜡包埋组织的使用受到限制。为了解决这个问题,发展了冷冻微阵列的方法。该方法快速便捷,效率很高,为使用临床标本及其相关的病理数据库来进行基因和蛋白质水平的研究提供了机会。该阐述了来自于组织和细胞系的微阵列的制备方法和技术特点,并概述它们在临床研究中的应用以及发展前景。     

微重力组织工程的产生与发展

概述了微重力组织工程的产生与发展趋势。动物细胞培养技术为研究细胞的形态、结构、功能与遗传特性 ,揭示细胞分裂、组织分化、器官组织形成等生命科学领域的基本问题发挥了巨大作用 ;大规模动物细胞培养技术已广泛应用于生物制药、生产疫苗以及生物学诊断试剂。但是 ,常规培养技术无法实现由细胞重建组织这一构想。微重力组织工程利用旋转培养器产生的微重力环境与动物细胞培养技术相结合 ,使组织重建成为可能 ,成为生命科学发展史上的一个新的里程碑。     

卵巢组织冻存与移植生育力保存的应用现状与研究进展

年龄增长、疾病以及抗肿瘤治疗等均可能会造成女性生育力的下降甚至丧失。随着诊疗技术的发展,癌症死亡率逐渐下降,因此患者的生育力保存需求较前增加。胚胎冻存、卵母细胞冻存和卵巢组织冻存是目前常用的女性生育力保存方式。卵巢组织冻存与移植至今仅有20余年历史,现已成为临床上具备医学指征的生育力保存方式,有助于重建患者的内分泌和生育功能,已有超200多例患者通过此技术成功完成生育。为提高冻存卵巢组织再移植的安全性,现有研究将卵母细胞体外成熟技术、人工卵巢技术与之相结合,避免携癌风险高的冻存卵巢组织移植后的癌症复发风险。较多的研究均已证实了卵巢组织冻存与移植技术的有效性和安全性。尽管卵巢组织冻存与移植技术得到了快速发展,但是该技术仍面临如何通过减少卵泡储备损失进而延长卵巢组织移植后有效性等挑战,尚需要更多的基础和临床研究促进卵巢组织冻存和移植技术的进步与发展。     

细胞膜片技术在组织工程中的应用

细胞膜片技术在组织工程中显示了良好的应用前景。与传统胰酶消化收集细胞的方法相比,运用该技术收集到的细胞保留了体外培养过程中分泌的胞外基质、建立的细胞-基质连接以及细胞间连接等结构。目前细胞膜片技术已被应用到角膜、食管、心脏、牙周韧带及软骨等多种组织修复中。然而,应用细胞膜片技术构建富血管化的大块组织、细胞膜片技术的工业化进程及基于细胞膜片技术的再生医学被大众所认知和接受仍是其在组织工程中面临的挑战。本文就细胞膜片技术在组织工程中的研究现状及最新进展作一综述,并寻求出其在临床及基础研究中的进一步发展方向。     

一种手工制作组织芯片技术的应用

组织芯片技术,是近年发展起来的一项以形态学为基础的生物学新技术,在实际工作中,我们在无专用制作芯片仪器的情况下,经多次反复实验找到了一种简单可行的制作组织芯片技术,经实践工作应用,取得了较好的效果,现介绍如下:     

组织工程皮肤发展现状

组织工程皮肤从概念提出至今技术发展迅速.本文对现有的组织工程皮肤进展展开论述,组织工程皮肤主要分3大类:由种子细胞和支架材料体外三维构建培养的组织工程皮肤、由细胞组成的组织工程化皮肤和由支架材料构成的组织工程化皮肤,根据其结构组成、形态或来源又分成2~3种,每种选1~3个代表具体描述.然后针对现有组织工程存在的再生修复性能不足、细胞来源受限、生产运输成本过高等技术问题进行分析讨论,同时就目前国家对该领域的管理办法进行了讨论和建议,并提出了组织工程皮肤的一些非移植性扩展应用.通过对组织工程皮肤领域技术成果的总结、技术问题与现有研究热点的讨论和未来前景的分析,希望能更好地促进该领域发展.     

自制组织芯片穿刺工具及芯片手工制作技巧

组织芯片（tissue chip TC）技术是将数十个乃至上千个的组织样本,整齐有序地排列在同一张载玻片上,而形成的微缩组织切片;又称组织微阵列（tissue microarray TMA）技术。组织芯片的制作方式主要有石蜡切片法和冷冻切片法,目前以石蜡切片法为常用。现在还有商品化的组织芯片制作仪,如美国（Tissue Arrayer,Beecher Instrument,USA）生产的,但这套产品价格昂贵。所以,手工制作技术更经济、实用,其方法的改进层出不穷。我们采用一次性穿刺针手工制做穿刺工具,用普通石蜡包埋样本,制做组织芯片,收到了满意的效果,现将制做主要程序和技巧介绍如下。     

快速成形技术制造组织工程支架研究进展

支架作为组织工程的关键要素之一, 影响着所接种细胞的分布和增值以及新组织的形成。传统的方法虽然可以制造出各种孔隙率的支架, 但缺乏对支架多孔结构的控制。近年来, 快速成形技术发展迅速, 并成功应用于组织工程支架的制造, 实现了组织工程支架内部多孔结构与复杂外形的精确控制, 从而使得构建理想的组织工程化结构体成为可能。以下回顾了应用快速成形技术制造组织工程支架的优势与潜力, 展望了未来组织工程支架的设计制造发展方向。     

应用于组织工程支架制备的电纺技术

组织工程技术为修复病损的组织和器官提供了一种新的途径,在组织工程中,细胞支架起着支撑细胞生长、引导组织再生、控制组织结构和释放活性因子等作用。针对电纺技术的新发展和细胞支架的新理念,综述了国内外利用电纺技术制备细胞支架的工艺条件、制备方法、组织细胞培养等方面的研究进展,并结合作者所在研究团队的研究工作提出了对未来电纺技术在组织工程中应用的研究重点和发展方向的认识。     

小鼠不同组织的差异线粒体蛋白质组研究

线粒体是真核细胞的重要细胞器, 它不仅具有合成ATP等普遍性功能, 也具备一定的组织特异性, 适应不同组织的生理功能需要. 为了鉴定组织相关性线粒体蛋白, 系统地分析和比较了C57BL/6J小鼠肝脏、肾脏和心脏线粒体蛋白质组. 通过双向凝胶电泳和MALDI-TOF/TOF质谱, 在这3种小鼠组织中共发现了87种组织特异的线粒体差异蛋白; 采用ICPL的定量蛋白质组方法和Western blotting技术, 进一步证实某些典型的组织特异性蛋白. 还通过实时定量PCR和Western blotting技术分析了6种具有代表性的组织差异线粒体蛋白质在线粒体内外的分布, 并结合激光共定位技术观察它们在相应组织活细胞内的表达与定位. 研究发现, 这些核基因编码的线粒体蛋白确有组织特异性分布和线粒体内外丰度差异特性. 本结果为组织特异性线粒体蛋白质的深入研究奠定了实验基础.     

Cuban Calisto (Lepidoptera, Nymphalidae, Satyrinae), a review based on morphological and DNA data

The Cuban species of Calisto are reviewed based on the morphology of adult and immature stages, as well as DNA sequences of six genes (COI, EF1α, wingless, GAPDH, RpS5, CAD). A new species, Calisto occultasp. n., is described from the northeastern Cuban mountains. Calisto smintheus Bates, 1935 and Calisto bruneri, Michener 1949 are revised and revalidated. A new status, the species level, is proposed for Calisto brochei, Torre 1973, Calisto muripetens, Bates 1939 and Calisto bradleyi, Munroe 1950. The immature stages of Calisto smintheus, Calisto brochei,and Calisto occulta are described for the first time, and those of Calisto herophile, Hübner 1823 are redescribed. Useful morphological characters for adults are the shape and conspicuousness of androconial patch, the number and relative size of white dots on underside of hindwing, the shape of aedeagus, the shape of digitiform projection of genitalia valve, the shape and relative size of tegumen and uncus, the relative size of female genitalia, the height of sterigmal ring dorsal crown of the latter, and the relative size of corpus bursae and ductus bursae. For the immature stages, the most important characters are the color pattern of head capsule, the number and width of longitudinal lines of body, in the larvae; and the color pattern and the absence or presence of dorsal ridges on the abdomen of pupae. The phylogenetic relationships between the Cuban Calisto species are quite robust and well-supported; however, conflict between mitochondrial and nuclear datasets was detected in Calisto brochei, Calisto muripetens and to a lesser degree in Calisto bradleyi.     

Areography of the Gymnosperms of China (1) Distribution of the Pinaceae of China

Nine of 10 genera and 119 of approximately 240 species of the Pinaceae occur in China, including 67 endemic species and two endemic genera. In this paper, the distributional maps of all the genera of the Pinaceae are presented (fig. 1-8). The horizontal and vertical distributions of species in each genus are discussed. The analysis of the distribution patterns of the genera indicates that some genera, such as Keteleeria, Tsuga, Pseudotsuga, Cathaya and Pseudolarix, are restricted to the area south of the Qinling Mountains and the Huaihe River, and the others, i. e. Picea, Abies, Larix and Pinus, extend northward to northeastern China. However, all of the genera except Keteleeria and Pinus are not found in very dry areas and tropical mountainous regions of China. The monotypic genera, Cathaya and Pseudolarix, are distributed in eastern and central China. The genus Keteleeria consists of 10 species, 7 of which are concentrated in southern Guizhou, northern Guangxi, southwestern Hunan and easternmost Yunnan. The distribution of the remaining 6 genera shows the maximum concentration in western Sichuan and northwestern Yunnan. (Figs. 2-8). Furthermore, more than third of species of the Pinaceae (37.8%) are also concentrated in western Sichuan and northwestern Yunnan. where a great variety of habitats and different topographic features occur. It is apparent that to conduct our systematic and evolutionary studies on this family in these region is especially needed. The relations between the areal size and the tolerance of species are discussed. The distributions of macrofossils and microfossils of the genera of the Pinaceae ia China are given, and it has been proved that areas of most genera of the family were considerably larger in the past. than at present.     

Origin,Differentiation, and Geographic Distribution of the Chenopodiaceae

Numbers of species and genera,endemic genera,extant primitive genera,relationship and distribution patterns of presently living Chenopodiaceae(two subfamilies,12 tribes,and 118 genera)are analyzed and compared for eight distributional areas,namely central Asia,Europe,the Mediterranean region,Africa,North America,South America, Australia and East Asia． The Central Asia,where the number of genera and diversity of taxa are greater than in other areas,appears to be the center of distribution of extant Chenopodiaceae．North America and Australia are two secondary centers of distribution． Eurasia has 11 tribes out of the 12,a total of 70 genera of extant chenopodiaceous plants,and it contains the most primitive genera of every tribe． Archiatriplex of Atripliceae,Hablitzia of Hablitzeae,Corispermum of Corispermeae,Camphorosma of Camphorosmaea,Kalidium of Salicornieae,Polecnemum of Polycnemeae,Alexandra of Suaedeae,and Nanophyton of Salsoleae,are all found in Eurasia,The Beteae is an Eurasian endemic tribe,demonstrating the antiquity of the Chenopodiaceae flora of Eurasia．Hence,Eurasia is likely the place of origin of chenopodiaceous plants． The presence of chenopodiaceous plants is correlated with an arid climate．During the Cretaceous Period,most places of the continent of Eurasia were occupied by the ancient precursor to the Mediterranean,the Tethys Sea．At that time the area of the Tethys Sea had a dry and warm climate．Therefore,primitive Chenopodiaceae were likely present on the beaches of this ancient land．This arid climatic condition resulted in differentiation of the tribes Chenopodieae,Atripliceae,Comphorosmeae,Salicornieae,etc．,the main primitive tribes of the subfamily Cyclolobeae. Then following continental drift and the Laurasian and Gondwanan disintegration, the Chenopodiaceae were brought to every continent to propagate and develop, and experience the vicissitudes of climates, forming the main characteristics and distribution patterns of recent continental floras. The tribes Atripliceae, Chenopodieae, Camphorosmeae, and Salicornieae of recent Chenopodiaceae in Eurasia, North America, South America, southern Africa, and Australia all became strongly differentiated. However, Australia and South America, have no genera of Spirolobeae except for a few maritime Suaeda species. The Salsoleae and Suaedeae have not arrived in Australia and South America, which indicates that the subfamily Spirolobeae developed in Eurasia after Australia separated from the ancient South America-Africa continent, and South America had left Africa. The endemic tribe of North America, the tribe Sarcobateae, has a origin different from the tribes Salsoleae and Suaedeae of the subfamily Spirolobeae. Sarcobateae flowers diverged into unisexuality and absence of bractlets. Clearly they originated in North America after North America had left the Eurasian continent. North America and southern Africa have a few species of Salsola, but none of them have become very much differentiated or developed, so they must have arrived through overland migration across ancient continental connections. India has no southern African Chenopodiaceae floristic components except for a few maritime taxa, which shows that when the Indian subcontinent left Africa in the Triassic period, the Chenopodiaceae had not yet developed in Africa. Therefore, the early Cretaceous Period about 120 million years ago, when the ancient Gondwanan and Laurasian continents disintegrated, could have been the time of origin of Chenopodiaceae plants.The Chinese flora of Chenopodiaceae is a part of Chenopodiaceae flora of central Asia. Cornulaca alaschnica was discovered from Gansu, China, showing that the Chinese Chenopodiaceae flora certainly has contact with the Mediterranean Chenopodiaceae flora. The contact of southeastern China with the Australia Chenopodiaceae flora, however, is very weak.     

Importance of biological and abiotic factors for geochemical cycling in a freshwater eutrophic lake

Here we report the results of a comprehensive biogeochemical monitoring of Rostherne Mere in 1998, including changes in dissolved oxygen, organic carbon and nitrogen, nitrate/nitrite, ammonia, Al, Na, S, K, Mg, Ca, Si, Fe, Mn, orthophosphate, particulate N & P, suspended solids, temperature, pH, chlorophyll-a and zooplankton. The results demonstrated the major influence of primary producers on the overall geochemical cycling of N, P and Si, and suggested that the significance of zooplankton might have been previously underestimated. For major anions and cations, however, the influence of biota on lake water concentrations appeared to be negligible, reflecting the fact that these chemicals were present far in excess of plankton requirements. Thus changes in concentrations of Ca, K, Na, Mg and S were rather limited and must have reflected changes in hydrological and meteorological parameters. K, however, demonstrated a transitional pattern, reflecting some influence of biological uptake. During the stratification period, the slow processes of bacterial decomposition in the hypolimnion gradually released chemicals contained in the materials accumulated in the bottom layer, remarkably increasing the concentrations of dissolved compounds of those elements present in amounts comparable with the pool stored in the sedimenting detritus (e.g. orthophosphate P, ammonia N, Si and DOC). The decomposition also resulted in a drop in the redox potential, followed by partial denitrification and chemical release from the sediments. The hypolimnion of the Mere was confirmed to remain at the stage of Mn release, characterised by accumulation of DOC, orthophosphates, ammonia and initial stages of denitrification. High levels of P released from the sediments during the stratification period suggest that the lake’s recovery after sewage diversion might be further delayed.     

Small Subunit rRNA Phylogenies Show that the Class Nassophorea is Not Monophyletic (Phylum Ciliophora)

ABSTRACT. The hypostome ciliates have been generally classified into two classes, Phyllopharyngea and Nassophorea. The status of Nassophorea and its relationship with Phyllopharyngea is one of the most controversial issues in ciliate systematics. Here we focus on the phylogenetic interrelationships of Nassophorea and Phyllopharyngea based on small subunit ribosomal RNA gene sequences. The three nassophorean subgroups, synhymeniids, microthoracids, and nassulids, each emerged as monophyletic, with synhymeniids as a sister group of Phyllopharyngea, and microthoracids as a sister of the synhymeniids+Phyllopharyngea clade in all phylogenies. The exact placement of the nassulids, however, remains uncertain. Following a detailed analysis of phenotypic characters, we hypothesize that: (1) the Phyllopharyngea could have evolved from synhymeniids, with the further development of their subkinetal microtubules as one of the major events; and (2) the development of monokinetid structures, as well as the reduction and specialization of the cyrtos and cortex, might have occurred during the diversifications of the microthoracids, synhymeniids, and Phyllopharyngea from a common ancestor. Expanding the class Phyllopharyngea to include the synhymeniids as a subclass, and designating a new subclass Subkinetalia n. subcl. for the group comprising cyrtophorians, chonotrichians, rhynchodians, and suctorians, are proposed.     

Origin and Evolution of Rosaceae

Rosaceae. consisting of about 126 genera and 3200 species, is widely distributed in warm temperate and subtropical regions of the Northern Hemisphere, while more than half of the genera are Asiatic and more then 80% of the total number of Asiatic occur in China (Table 1). In this paper, the origin and evolution of Chinese genera is discussed mainly. The principal tendency of the whole family is also described from the point of view of evolution. First of all, the systematic position of Rosaceae in Angiospermae is reviewed. According to the records of paleobotany, rosaceous plants occurred first in the Tertiary, from the early period of Eocene (genera such as Spiraea and Prunus) to the late period of Miocene (e.g. Crataegus, Malus amd Rosa). They have quite a long history in geological data. Where has this big and old family originated and what steps does it stand in the long course of evolution of flowering plants? There are several opinions and explanations by different authors. In this paper, a general survey of the six prevailing classical systems (Table 2) is made to give a brief idea of the position of this family in the Angiospermae and of the relationships between the subfamilies and also the relationships between different genera in each subfamily. At the end of this paper, an attempt is made to analyse and sum up the major evolutionary tendency of the whole family. As generally condidered, Rosaceae originated from Magnoliales, and woody plants of the family still hold a dominant position. For instance, subfamily Spiraeoideae consists of only one herbaceous genus (i.e., Aruncus) and subfamily Rosoideae only a few herbaceous genera. All of these herbaceous genera are derived from the closely related woody genera of the same subfamily. In the course of evolution of Angiospermae, Rosaceae stands at the initial to the middle stages of development. All parts of plant body in this family are at the chang ing and developing stages, with carpels, fruits and inflorescences being the most active. The primitive types in this family, such as the members of subfamily Spiraeoideae, usually have 5 and free carpels, the number of which are either reduced to 2-1 or increased to 10-numerous. They have different levels of union and are either completely free from each other or coherent at base. The carpels usually occur on the upper part of the receptacle, because the shapes of receptacle are variable, sometimes disk-shaped, cupshaped, tube-shaped or even bottle-shaped. In the last case carpels grow inside the receptacle. Thus the position of carpels has changed from superior to inferior through halfsuperior. In accordance with the development of the carpels, various kinds of fruits are produced. The primitive types of fruit are follicles, with dry, dehiscent carpels opened along different sutures. The next step, the carpels have developed into an indehiscent, I-celled and l-seeded fruit, the so-caned achene. In different genera, the achenes have different coat types and appendages to facilitate dispersing the seeds. Some of the achenes grow upon the fleshy receptacle (like strawberry) and some of them inside the fleshy receptacle (like rose). Sometimes a few carpels are united with the receptacle and develop into a pome (like apple and pear). Another direction of the fruit development is the single carpel with fleshy exocarp and mesocarp, and a bony endocarp, then becoming a drupe (like peach and plum). In addition to fleshy receptacle of thickened fruit coats, they usually have showy colour, fragrant smell and also plenty of sugars, acids, vitamins, etc. which are edible and attract animals and human beings to assist the dispersion of seeds. In this family, there are various types of flower arrangements, both indefinite inflorescences including raceme, umbel, corymb and panicle, and the definite inflorescence, such as solitary flower, cyme and compound cyme. In the evolution course, they tend to change mostly from multiflowered compound inflorescence towards few-flowered simple inflorescence, and finally becoming a solitary flower: simultaneously with the decreasing of number of flowers on the inflorescence, the increasing of size of petals, which become very showy for attraction of insects so as to guarantee pollination and fertilization of the plants concerned. Another tendency, if the bisexual flowers change to unisexual, either monoecious- or dioecious-polygamous, then they form a dense spike which is beneficial to cross pollination. The abundance, diversity, and wide range of distribution of the species and genera of Rosaceae are considered mainly resulted from their highly developed reproductive organs.     

云南复叶耳蕨属的分类修订

研究了云南产复叶耳蕨属植物,提出了新的分类处理。云南共有本属植物２０种２变种,其中包括１新组,３个改级新组合,７个云南分布新记录,并清理出了５４个云南及其他省区种类的新异名。     

草鱼鳃上寄生毛管虫一新种——变异毛管虫的研究

吸管亚纲(Subclass suctoria)中,多数种类具有或长或短的炳,附着它物上营固着生活。寄生在鱼类体表、鳃丝内的毛管虫(Trichophrya)和簇管虫(Erastophrya)的种类中,至今未见报道有固着柄的代表。       

The impact of European Directives on estuarine and coastal science and management

Many recent developments in European marine and estuarine science have been against the demands of European Union legislation. The implementation of certain statutes, the role of scientists and the nature of the data required are discussed using examples from the UK, the Netherlands and Portugal. This includes the implementation of the EU Directives on Urban Waste-water Treatment, the control of Nitrates, the designation of Species and Habitats, the control of Dangerous Substances, the statutory requirement for Environmental Impact Assessments and the recently proposed Water Framework Directive. For these, the integration of physical, chemical and biological monitoring and investigation is discussed in relation to the science dictated by the legislative and administrative requirements. Each of these Directives requires the development of generic guidelines and protocols for implementation and the use of national enabling legislation. This indicates that, in several cases, the science should concentrate on monitoring and assessment in a well-structured and quality-controlled manner. The paper concludes by summarising developments based on similarities in the implementation of present and proposed Directives across Europe.