端粒酶RNA反义基因对肝癌细胞的影响

用RT-PCR的方法钓取端粒酶RNA基因的cDNA,并将其反向插入到逆转录病毒载体pLNCX上,构建hTR基因的反义表达质粒。将质粒经脂质体介导转染人肝癌SMMG-7721细胞中表达。结果表明hTR反义基因的表达有效地封闭或抑制肝癌细胞的端粒酶活性,抑制细胞的生长和增殖,延长细胞的倍增时间并促进细胞凋亡。     

胡桃楸提取液对肿瘤细胞增殖的抑制作用

目的考察胡桃楸提取液对肿瘤细胞Hela、K562的抑制作用和相关机制。方法用MTT方法分析胡桃楸提取液对Hela、K562细胞增殖的影响。采用端粒酶PCR ELISA试剂盒分析胡桃楸提取液对Hela、K562细胞端粒酶的影响。结果 Hela细胞24、48和72 h的LD50分别为406.18μg/mL、319.48μg/mL和112.84μg/mL。K562细胞24 h LD50为154.50μg/mL。HLF细胞LD50为918.69μg/mL。胡桃楸提取液可抑制Hela细胞和K562细胞的端粒酶活性,而对HLF细胞端粒酶活性影响不大。结论胡桃楸提取液对Hela细胞、K562细胞有抑制作用,在低浓度下对HLF细胞杀伤不大。对肿瘤细胞抑制作用可能与抑制端粒酶活性相关。     

浑河流域2010-2014年的鱼类群落和水生态健康变化分析

为了解浑河流域鱼类群落结构的变化趋势和水生态健康变化,分别于2010年和2014年对浑河流域46个采样点进行了鱼类和环境因子调查。其中2010年采集到鱼类15039尾,隶属于6目9科32种;2014年采集到鱼类10483尾,隶属于6目10科41种。Mann-Whitney U检验表明从2010年到2014年鱼类总物种数、香农多样性指数、底层物种数百分比、植食性和肉食性个体数百分比等鱼类特征参数显著上升。典型对应分析（CCA）结果表明,2010年显著影响鱼类群落结构的环境因子为流速、电导率、河流等级和钙离子浓度;而2014年显著影响鱼类群落结构的环境因子则为电导率、河流等级和氨氮浓度。2010年和2014年的鱼类完整性指数（FIBI）评分分别为48.7624.82和50.4117.35,与2010年的评价结果相比,2014年F-IBI结果稍有改善,其中极好和好的点位数分别少3个和1个,一般的点位数多9个,而极差和差的点位分别少4个和1个。     

荧光定量PCR 法检测肿瘤细胞端粒酶活性的变化

目的：利用荧光定量PCR法检测端粒酶抑制剂作用于人肝癌细胞SMMC-7721后端粒酶活性的变化,探讨其抑制端粒酶活性的可能机制,为端粒酶抑制剂的临床应用提供理论依据。方法：利用荧光染料SYBR—Green I建立一种新的端粒酶活性检测方法：FQ—TRAP法。利用FQ—TRAP法检测端粒酶抑制剂作用后肿瘤细胞端粒酶活性变化。结果：端粒酶抑制剂作用后,肝癌细胞端粒酶活性都有变化,其中以ASODN,EGCG,AZT抑制效果较明显。结论：端粒酶FQ—TRAP法是一种特异性、灵敏度、重复性都较好,可快速、简便及定量检测人端粒酶活性的方法,端粒酶抑制剂作用后癌细胞端粒酶活性的变化,为端粒酶抑制剂的临床应用提供理论依据。     

上海苏州河治理中鱼类多样性及群落结构变化

为了评估近20年来上海苏州河环境综合整治过程中鱼类多样性的变化状况, 本文于2019年6月(夏季)和9月(秋季)对苏州河13个断面的鱼类样本进行了采集, 对鱼类多样性和群落结构作了分析。结果显示, 2次采集共获得鱼类样本10,102号, 隶属于8目15科37属45种。夏季和秋季, 上游8个断面的鱼类均为36种, 下游5个断面则为12种和15种。从上游到下游, Shannon-Wiener多样性指数(H')、Margalef种类丰富度指数(D)和Simpson优势度指数(C)总体呈逐渐降低的趋势。相对重要性指数(IRI)显示, 夏秋两季上游断面共同的优势种为似鳊(Pseudobrama simoni), 下游为鲫(Carassius auratus)和泥鳅(Misgurnus anguillicaudatus)。丰度生物量曲线表明, 全河段鱼类群落结构总体上并不稳定, 特别是夏季的下游河段。Cluster聚类和非参数多维标度排序分析可明显将鱼类群落分为上游群组(I)与下游群组(II), 似鳊、泥鳅、鲫、光泽黄颡鱼(Pelteobagrus nitidus)和兴凯鱊(Acheilognathus chankaensis)等是造成群组间差异性的主要分歧种。与2001年的43种、2006年的28种相比, 本次调查的物种数出现了明显回升, 特别是下游河段。这种上游鱼类向下游迁移的趋势, 预示着下游中心城区与上游郊区河段的水质差别正在缩小, 水生态系统恢复迹象明显。本文认为, 上下游之间鱼类群落结构差异的缩小以及翘嘴鲌(Culter alburnus)、红鳍原鲌(Cultrichthys erythropterus)、棒花鱼(Abbottina rivularis)、子陵吻虾虎鱼(Rhinogobius giurinus)等在下游河段的出现, 可作为苏州河综合整治效果的重要生态标志。     

Telomeres,senescence, and hematopoietic stem cells

The replicative lifespan of normal somatic cells is restricted by the erosion of telomeres, which are protective caps at the ends of linear chromosomes. The loss of telomeres induces antiproliferative signals that eventually lead to cellular senescence. The enzyme complex telomerase can maintain telomeres, but its expression is confined to highly proliferative cells such as stem cells and tumor cells. The immense regenerative capacity of the hematopoietic system is provided by a distinct type of adult stem cell: hematopoietic stem cells (HSCs). Although blood cells have to be produced continuously throughout life, the HSC pool seems not to be spared by aging processes. Indeed, limited expression of telomerase is not sufficient to prevent telomere shortening in these cells, which is thought ultimately to limit their proliferative capacity. In this review, we discuss the relevance of telomere maintenance for the hematopoietic stem cell compartment and consider potential functions of telomerase in this context. We also present possible clinical applications of telomere manipulation in HSCs and new insights affecting the aging of the hematopoietic stem cell pool and replicative exhaustion. This work was supported by European Community Grant LSHC-CT-2004-502943 (MOL CANCER MED).     

Immortalization with telomerase of the Nestin-positive cells of the human pancreas

Cells expressing the neuronal stem cell marker Nestin are present in the human pancreas but the biological role of these cells has yet to be resolved. We report here the establishment with the catalytic subunit of human telomerase (hTERT) of a line of normal human cells representing this cell type. Primary human cells derived from the ducts of the pancreas were transduced with an hTERT cDNA. The infected cells became positive for telomerase, failed to senesce, and were still proliferating after more than 150 doublings. The immortalized cells were positive for the expression of Nestin (at both the mRNA and protein levels) and were found to be free of cancer-associated changes: diploid and expressing wild type p16(INK4a), p53, and K-Ras. An established line of normal human cells representing this cell type should be of great value to help define the biological properties of this novel cell type.     

The role of farm structure on bird assemblages around a Kenyan tropical rainforest

Although it is clear that the farmlands neighbouring fragmented forests are utilized by some forest birds, it is not clear how birds in general respond to farmland habitat mosaic. An effort was made to determine how bird density and foraging assemblages were influenced by farm structural characteristics and distance from forest edge. Thirty farms up to a distance of 12 km around Kakamega forest in western Kenya were studied. Farm structure entailed size, hedge volume, habitat heterogeneity, woody plant density, plant diversity and crop cover. Birds were surveyed using line transects and DISTANCE analyses and classified into six feeding guilds and three habitat associations. Size of farms increased away from the forest, as woody plant density, plant diversity, indigenous trees and subsistence crop cover declined. The most important farm structure variable was hedge volume, which enhanced bird species richness, richness of shrub‐land bird species and insectivorous bird density (R = 0.58, P < 0.01). Bird density increased with tree density while indigenous trees were suitable for insectivores and nectarivores. There were very few forest bird encounters. Agricultural practices incorporating maintenance of hedges and sound selection of agroforestry trees can enhance conservation of birds on farmland, though, not significantly for forest species.     

Synthesis of combretastatin A-4 O-alkyl derivatives and evaluation of their cytotoxic,antiangiogenic and antitelomerase activity

We here report the synthesis and biological evaluation of several combretastatin A-4 derivatives alkylated at the phenol hydroxyl group. Some of these derivatives contain an (E)-arylalkene fragment reminiscent of that present in some natural stilbenes like resveratrol. The cytotoxicities towards one human healthy kidney embryonic and two tumoral cell lines were determined. In addition, the ability of these compounds to inhibit the production of the vascular endothelial growth factor (VEGF) was measured. Finally, the expression of genes controlling the production of telomerase was measured. Some of the compounds were found to have an activity comparable or higher than that of combretastatin A-4 in at least one of the aforementioned biological properties. The compounds with the (E)-arylalkene fragment were in general terms more active than the simple O-alkyl derivatives. However, no clear structure/activity correlations were perceived when comparing the observed compound activities across the three biological properties. This points out the existence of marked differences between the mechanisms responsible for their cytotoxicity.     

Stream fish communities and their associations to habitat variables in a rain forest reserve in southeastern Brazil

This paper describes the spatial variability of fish communities and identifies patterns of association between fish communities and habitat variables, including anthropogenic factors. We sampled streams inside and in the surroundings of a rain forest reserve in the southeast of Brazil in the rainy season. We could distinguish three main groups of streams: upland streams (draining the upland, flat portions of the mountain ridge of Serra de Paranapiacaba), adventitious streams of clearwater mountain torrents (small streams draining confined valleys in the slopes of this sierra), and large streams of clearwater mountain torrents (relatively unconstrained large streams close to the foots of the mountains). Despite the high variability of fish communities associated with these streams we identified some patterns using exploratory statistical analyses. These patterns were corroborated by additional field observations and information from the scientific literature. The main differences in fish community composition and diversity among the three groups of streams are probably related to large-scale factors such as elevation and position of the stream in the watershed. However, differences within these three groups seem to be mostly due to site-specific factors. Differences of instream characteristics are likely to be caused by natural variability of the ecosystems but also, in some cases, by human disturbances like pollution from human settlements, agriculture and mining.