中国栅蛛属2新种记述(蜘蛛目:栅蛛科)

本文记了分别采自云南高黎贡山的栅蛛科栅蛛属Hahnia 2新种:垭口栅蛛,新种S.yakouensis sp.nov.和肾形栅蛛,新种S.reniformis sp.nov..垭口栅蛛后眼列前曲,交媾腔大,扁圆形,交媾孔1个,位于交媾腔下缘,交媾管粗,呈"人"字形下行分成2支再向两侧扭曲.纳精囊有一肓管斜向上伸出,鉴于上述特征而与Hahnia mridulae Tikader,1970不同.肾形栅蛛交媾孔2个,位于生殖厣腹面中央,纳精囊1对,大,肾形,插入器始于生殖球左下方,鉴于上述特征而与Hahnia xinjiangensis Wang et Liang,1989不同. Abstract： The present paper deals with two new species of the genus Hahnia collected from the Gaoligong Mountains Region of Yunnan Province, China: Hahnia yakouensis sp. nov., Hahnia reniformis sp. nov..     

注重发挥博士后科研生力军作用

中国医科大学博士后科研流动站始建1995年。目前学校设有基础医学、临床医学和生物学3个博士后科研流动站。但由于每年国家财力有限,资助名额很少,在有限条件下建立、健全博士后制度,加强对博士后人员的培养,调动博士后人员的积极性是一项重要的研究课题。     

遗传物质的发现者之一——麦卡锡

1944年,3位科学家艾弗里、麦卡锡和麦克劳德在DNA遗传本质方面的发现是20世纪最重要的发现之一,这个发现打开了生物学革命的大门,从而改变了人类对自然界的看法,这项研究还为1953年沃森和克里克DNA双螺旋结构的发现奠定了坚实的基础,但不幸的是3位科学家都未曾荣获诺贝尔奖.通过介绍麦克林·麦卡锡的科学研究,从而对这项发现的基本状况有一个基本的了解.     

生命之树与进化模式:导言

One and half centuries ago, Charles Darwin (1859) presented overwhelming evidence and argued that all life on the earth shared common descent, and "from so simple a beginning endless forms most beautiful and most wonderful have been, and are being evolved". Ernst Haeckel (1886) and several of his contemporaries attempted to trace the pattern of descent among all extant and extinct forms in what Darwin referred to as "the great Tree of Life". Ever since then, systematists and evolutionary biologists have been exploring morphological, cytogenetic, chemical, developmental and molecular characters, and actively developing theories and methods to infer phylogenetic relationships among organisms from these characters. This endeavor has been especially stimulated by the rise of molecular biology and the emergence of computer science over the past 50 years. At the beginning of the 21st century, we are presented with an unprecedented opportunity to reconstruct the entire Tree of Life, and further, to study evolutionary processes and mechanisms in the context of a robust phylogenetic framework.     

我国微生态学的奠基人——魏曦

魏曦,字东升,1903年12月25日出生于湖南岳阳一个小职员家庭,父亲任职于邮政局.1914～1921年他在家乡湖滨中学读书,毕业后考入长沙湘雅医学院,学习两年后曾参加北伐军,任第四集团军警卫团三等军医.后退出军队,在长沙广雅中学任教.1928年入设立在上海的中央大学医学院(1932年独立为上海医学院)学习,1933年毕业,获博士学位.     

高黎贡山北段东西坡种子植物区系的比较研究

高黎贡山北段的东西坡由于在降雨量和热量分配等方面存在着显著的差异,致使东西坡在植物的种类、组成及区系特征等方面表现出明显的差异.东坡记载野生种子植物152科,580属,1475种及192变种(亚种),西坡记载野生种子植物162科,659属,1804种及186变种(亚种).东西坡种子植物科、属、种的对比分析表明:1)东西坡现代种子植物区系具有相同的历史渊源,但其区系联系减弱了,东西坡区系相似性程度,依科、属、种的顺序依次递减;2)西坡现代种子植物区系比东坡具有更为深刻的热带起源烙印.就科、属、种三个水平来说,东坡的热带成分低于西坡,温带成分高于西坡.许多典型的泛热带大科在西坡比东坡有着更为丰富的种类,其中有些泛热带科分子在东坡缺乏分布,而在西坡找到了合适的驻留之地;3)西坡现代种子植物区系与东喜马拉雅植物区系的联系比东坡紧密,而东坡与高黎贡山以东的区系联系比西坡密切,由于高黎贡山山脉的阻隔,近代植物物种的东西坡交流发生了障碍;4)西坡生态地理环境比东坡更有利于物种的生存、繁衍和分化,它既是古老成分的避难所,又是孕育新生成分的摇篮.     

用叶绿体ndhF和核核糖体ITS序列推断李属(蔷薇科)的系统发育

Sequences of the chloroplast ndhF gene and the nuclear ribosomal ITS regions are employed to reconstruct the phylogeny of Prunus (Rosaceae), and evaluate the classification schemes of this genus. The two data sets are congruent in that the genera Prunus s.l. and Maddenia form a monophyletic group, with Maddenia nested within Prunus. However, the ndhF data set is incongruent with the ITS data supporting two major groups within Prunus one consisting of subgenera Laurocerasus (including Pygeum) and Padus as well as the genus Maddenia and another of subgenera Amygdalus, Cerasus, and Prunus. The ITS data, on the other hand, support a clade composed of subgenera Amygdalus and Prunus and Prunus sect. Microcerasus in addition to a paraphyletic grade of subgenera Laurocerasus and Padus (and the genus Maddenia) taxa. In general, the subgeneric classifications of Prunus s.l. are not supported. The ITS and ndhF phylogenies differ mainly in interspecific relationships and the relative position of the Padus/Laurocerasus group. Both ITS and ndhF data sets suggest that the formerly recognized genus Pygeum is polyphyletic and that the distinction of the subgenera Padus and Laurocerasus is not supported. The biogeographic interactions of the temperate and tropical members in the Padus/Laurocera- sus/Maddenia alliance including Pygeum are shown to be highly dynamic and complex.     

途经甘肃的3种水鸟新纪录

2007年10月13日至11月5日进行敦煌市湿地鸟类调查时,分别在南湖湿地与候鸟自然保护区及党河水库使用Leica apo77高倍望远镜观察到3种水鸟,在以往的文献资料中未见其分布于甘肃的报道,应为甘肃鸟类新纪录。笔者用500mm镜头分别拍下3种水鸟的照片。1.赤颈(Podiceps grisegena)2007年10月13日13:30时,在南湖湿地与候鸟自然保护区的阳关水库(渥洼池)记录到2只赤颈。当时水面上同时有凤头(P.cristatus)、黑颈(P.nigricollis)以及大量鸭类、潜鸭类游禽活动,赤颈体形较凤头小,而又明显比黑颈大。其中一只赤颈的…     

鸡的瞬膜

瞬膜(nictitating membrane),又称第三眼睑,是一种保护眼球、防止灰尘的结构.鸟类的瞬膜位于眼眶的前眼角,为半透明的膜,其内缘具有一种羽毛上皮,借以刷洗角膜上的灰尘.在飞行时能遮覆眼球,以避免干燥气流和灰尘对眼球的伤害.由于瞬膜在鸟类睁眼的一瞬间迅速缩回前眼角,很难拍摄到.最近费了好大的周折,终于拍到了理想的鸡瞬膜照片,现予以发表供生物学界的同行共享(友情提示:如引用请注明原作者).     

Morphological selection of parental Chinese hamster ovary cell clones exhibiting high-level expression of recombinant protein

To facilitate the establishment of recombinant Chinese hamster ovary (rCHO) cell lines with dihydrofolate reductase (dhfr)-mediated gene amplification, a primary selection method based on morphology of parental CHO clones has been developed. Morphology of parental clones that were made by transfecting various plasmids encoding thrombopoietin (TPO) and its analogs and humanized antibodies into dhfr-deficient (dhfr-) CHO cells was not uniform. Morphology of many parental clones exhibiting high-level expression of the introduced gene was similar to that of nontransfected dhfr- CHO cells. On the other hand, most parental clones with low-level expression experienced noticeable morphological changes such as bipolar fibroblast-like morphology. In case of selection of parental clones with TPO expression level higher than 200 ng/mL, morphological selection improved selection efficiency by 3.5-fold compared with random selection. Furthermore, when subjected to methotrexate for gene amplification, parental clones that were selected based on morphology elevated the expression level as much as those that were selected randomly. Taken together, morphological selection of parental clones can facilitate the establishment of rCHO cell lines expressing recombinant proteins.     

Generation of recombinant CHO(dhfr-) cell lines by single selection for dhfr+ transformants.

In order to establish a mammalian cell expression system with a minimum of selection steps and a stable expression of microgram amounts of recombinant protein (human tissue-type plasminogen activator mutants and chimeric proteins) per 10(6) cells per day, we investigated Chinese hamster ovary cells and the dihydrofolate reductase-deficient Chinese hamster ovary cell line CHO(dhfr-). The 1tPA expression vector pCMVtPA was cotransfected either with the SV40 enhancer sequence containing dhfr expression vector pMT2 or with the enhancerless dhfr expression vector pAdD26SV(A) into CHO(dhfr-) cells. With both dhfr expression plasmids, selection for dhfr+ transformants followed by single dilution cloning was sufficient to generate cell lines with a production level of up to 4.6 micrograms tPA/10(6) cells.day. This approach is useful if gene amplification procedures are time-consuming and impracticable because of a large number of recombinant proteins. In order to establish CHO cell lines with a tPA expression level as high as that in the case of CHO(dhfr-) cells, repeated dilution cloning is necessary.     

Effects of synthetic lysophosphatidylcholines on suspension cultures of the Chinese hamster ovary DG44 cells in protein-free media

This study described the effects of synthetic lysophosphatidylcholines on the growth of recombinant CHO-DG44 cells in suspension. Overall, cell growth characteristics were improved when cultivated in suspension in a protein-free medium supplemented with natural soybean lysophosphatidylcholines. To substitute synthetic lysophosphatidylcholines for the naturally occurring lysophosphatidylcholines, we implemented a systematic approach in which twelve synthetic lysophosphatidylcholines were grouped into three lipid mixtures according to the length of their acyl chains. We found that synthetic lysophosphatidylcholines with medium acyl chain lengths (C14-C18), including oleoyl lysophosphatidylcholine (C18:1) could increase cell growth in the protein-free media. The fortified protein-free medium with medium acyl chain length lysophosphatidylcholines (C14-C18) maintained growth of CHO-DG44 cells over five consecutive passages, whereas the cell growth in a CHO protein-free medium was decreased gradually after four passages. We also observed that the restorative effect of oleoyl lysophosphatidylcholine was comparable to that of natural lysophosphatidylcholine in batch and long-term cultivation. These results show that synthetic lysophosphatidylcholines can be used as lipid supplements in either protein-free media or chemically defined media for CHO cell suspension cultures.     

Effects of insulin and LongR(3) on serum-free Chinese hamster ovary cell cultures expressing two recombinant proteins

Insulin is the most commonly used growth factor for sustaining cell growth and viability in serum-free Chinese hamster ovary (CHO) cell cultures. In the present study insulin and IGF-1 analogue (LongR(3)) were compared for their ability to support growth, viability, and production of two serum-free CHO cell lines expressing recombinant protein. The first cell line, VA12, expresses protein B, and the second cell line, CL23, expresses protein C. Both molecules are recombinant cytokine receptors. VA12 will grow in serum-free media lacking growth factor, while CL23 requires either insulin or LongR(3) for cell growth. Both cell lines, however, require a growth factor for optimal performance under production conditions. In this study, LongR(3) was better able to sustain the viability of both cell lines under production conditions than insulin. These data indicate that while insulin and LongR(3) can both serve as growth and viability factors for CHO cells, LongR(3) is the preferred growth factor for cell lines VA12 and CL23.     

Serum protects protein-free competent Chinese hamster ovary cells against apoptosis induced by nutrient deprivation in batch culture.

The development of serum- and protein-free Chinese hamster ovary (CHO) cell cultures is a high priority for the production of biopharmaceuticals. Protein-free competent CHO cells lines have been previously constructed by two different methods-metabolic engineering with cell-cycle regulatory proteins and long-term selective adaptation. Apoptosis was present in both cell lines during protein-free, static-batch culture as a result of nutrient deprivation, and glucose deprivation alone was a potent inducer of apoptosis compared to the depletion of other nutrients such as amino acids. By adding back serum to the cultures during batch growth or nutrient deprivation, it was shown that unidentified survival factors in serum can greatly reduce apoptosis in protein-competent cell lines in all phases of the culture. Both observations contrast to previous reports for hybridoma cells, in which amino acids were the key determinants of apoptosis and serum had no additional antiapoptotic effect. Serum's protective effect against CHO cell death in batch culture was multifaceted and complex: (1) 10% FBS increased cell viability to >99% during exponential growth from roughly 75-90%, (2) 5-10% fetal bovine serum (FBS) reduced specific glucose consumption rates in both cell lines by 40%, thereby delaying the onset of apoptosis caused by glucose deprivation, and (3) 5% FBS reduced the specific cell death rate by 65% during a 3-d lactate-consumption phase characterized by substantial abortive proliferation, in which the cells both proliferated and died at a constant rate. The benefit of serum on cell production over the various phases of batch growth was combined into a single parameter by integrating the viable cell concentration vs. time profile (termed here as cumulative volumetric viable cell-time, VCTvol). Despite the ability of both cell lines to grow indefinitely without any exogenous growth factors, the addition of serum resulted in a 2. 3-fold increase in the VCTvol. Thus, it is clear that there is much room for improvement of protein-free CHO cell lines despite their adequate growth competence, and new strategies different from those successfully used for hybridomas may be necessary to combat CHO cell apoptosis.     

Comparative Assessment on the Expression Level of Recombinant Human Follicle-Stimulating Hormone (FSH) in Serum-Containing Versus Protein-Free Culture Media

Production of recombinant pharmaceutical proteins has made a great contribution to modern biotechnology. At present, quick advances in protein expression lead to the enhancement of product quantity and quality as well as reduction in timescale processing. In the current study, we assessed the expression level of recombinant human follicle-stimulating hormone (rhFSH) in adherent and suspension Chinese hamster ovary (CHO) cell lines by cultivation in serum-containing and chemically defined, protein-free media. The expression cassette entailing FSH subunits was transfected to CHO/dhfr- and CHO DG44 cell lines, and gene amplification was achieved using dihydrofolate reductase (DHFR)/methotrexate (MTX) system. Afterward, the expression level of rhFSH was studied using real-time PCR, Western blotting and ELISA. Our achievements revealed that stepwise increase in MTX [up to 2000 nano-molar (nM)] leads to boost the expression level of rhFSH mRNA in both cell lines, although DG44 have better results, as mRNA expression level reached 124.8- and 168.3-fold in alpha and beta subunits, respectively. DG44 cells have also the best protein production in 2000 nM MTX, which reached 1.7-fold in comparison with that of the mock group. According to the above results and many advantages of protein-free media, DG44 is preferable cell line for future steps.     

Impact of coexpression and coamplification of sICAM and antiapoptosis determinants bcl-2/bcl-x(L) on productivity,cell survival,and mitochondria number in CHO-DG44 grown in suspension and serum-free media

We have engineered dihydrofolate reductase-negative (dhfr-/-) Chinese hamster ovary (CHO) DG44 cells adapted for growth in serum-free suspension cultures for simultaneous expression of the common cold therapeutic, the soluble intercellular adhesion molecule 1 (sICAM), and the antiapoptosis determinants bcl-2 or bcl-x(L). Detailed analyses of titer and antiapoptosis characteristics of these production cell lines included an independent (sICAM; bcl-2/bcl-x(L)) as well as a cocistronic (sICAM-(bcl-2/bcl-x(L))) expression set-up in which translation-initiation of the survival cistron is driven by an internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV). In transient transfections or stable mixed populations and in comparison to isogenic sICAM-only control vectors, both bcl-x(L)-encoding configurations achieved higher sICAM yields while bcl-2 over-expression resulted in decreased product levels. Overall, the death-protective impact of bcl-2 and bcl-x(L) in engineered CHO-DG44 was not significant under typical batch-mode operation, an observation that was confirmed by clonal analysis. bcl-2 and bcl-x(L) displayed their antiapoptosis potential only following dhfr-based amplification in sICAM-producing CHO-DG44 cell lines. In all cases, bcl-x(L) outperformed bcl-2 in its cell death-protective capacity. Amplification-dependent high-level expression of mitochondria-localized bcl-2 family members required for successful antiapoptosis engineering may be essential to compensate for increased mitochondria numbers found to be associated with production cell lines grown in serum-free medium.     

Reconstitution of functionally active antibody directed against creatine kinase from separately expressed heavy and light chains in non-lymphoid cells

We report here for the first time reconstitution and secretion of functionally active antibody in non-lymphoid cells. Expression vectors for the light and the heavy chain of a monoclonal antibody directed against creatine kinase (EC 2.7.3.2) were introduced into COS and CHO Chinese hamster ovary dhfr- cells. Introduction of the expression vectors separately gave rise to immuno-reactive material in the culture supernatants, but only cotransfection of the expression plasmids resulted in secretion of protein with immuno-reactivity against antibodies directed against mouse heavy and light chains as well as specific antigen-binding affinity, as determined by enzyme-linked immunosorbent assay. Secreted kappa and gamma chains from reconstituted antibody were characterized by immunoadsorption and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In COS cells, reconstituted antibody was transiently secreted; cotransfection of kappa and gamma chain expression plasmids with a dihydrofolate reductase (DHFR)-expression plasmid into CHO dhfr- cells gave rise to stable transformants secreting functionally active antibody.