人体上皮细胞系（株）间交叉污染的检测

本文根据不同上皮细胞的角蛋白丝性质和多肽组成的差异,建立了四种不同上皮细胞系(株)间交叉污染的检测方法:1.SDS-PAGE法;2.免疫印迹法;3.AE1单抗免疫荧光染色法;4.角蛋白丝结构转化法。结果表明:方法1—3比较有用。我们认为,要获得较满意的检测结果,需要根据不同上皮细胞的特点,选择不同的方法配合使用。     

Isozyme differentiation in the endemic genusArgyranthemum (Asteraceae: Anthemideae) in the Macaronesian Islands

The genusArgyranthemum (Asteraceae: Anthemideae) comprises 38 taxa and is restricted to the archipelagos of the Canaries, Selvagens, Madeira, and Desertas in the Macaronesian biogeographic region. An electrophoretic study, including 17 enzyme loci and at least one population of each of the described taxa, was carried out. High identity (low distance) values between taxa (mean of 0.893) were obtained despite the old age of the islands, their close proximity to the African continent, and the fact thatArgyranthemum is the most species-rich and variable genus in Macaronesia. These results suggest that the genus is monophyletic and that it has evolved very rapidly in these islands. There is little correspondence between taxonomy and neighbor-joining analysis based on Roger's genetic distances, but in several instances populations from the same islands cluster together despite being from different species or even different sections. It is suggested that repeated genetic bottlenecks associated with the founding of new populations during radiation of the genus resulted in lineage sorting of ancestral allozyme polymorphisms. Because every population has a high average identity with all other populations, lineage sorting could result in populations of different taxa being slightly more similar than populations of the same taxon. Gene flow between different species on the same island could account for some populations clustering by island of origin rather than taxonomic disposition. Average allozyme diversity within populations (0.098) is 50% higher than the mean total diversity for species endemic to oceanic islands.     

A scaling approach for quantifying the net CO2 flux of the Kuparuk River Basin,Alaska

Net CO₂ flux measurements conducted during the summer and winter of 1994–96 were scaled in space and time to provide estimates of net CO₂ exchange during the 1995–96 (9 May 1995–8 May 1996) annual cycle for the Kuparuk River Basin, a 9200 km² watershed located in NE Alaska. Net CO₂ flux was measured using dynamic chambers and eddy covariance in moist‐acidic, nonacidic, wet‐sedge, and shrub tundra, which comprise 95% of the terrestrial landscape of the Kuparuk Basin. CO₂ flux data were used as input to multivariate models that calculated instantaneous and daily rates of gross primary production (GPP) and whole‐ecosystem respiration (R) as a function of meteorology and ecosystem development. Net CO₂ flux was scaled up to the Kuparuk Basin using a geographical information system (GIS) consisting of a vegetation map, digital terrain map, dynamic temperature and radiation fields, and the models of GPP and R. Basin‐wide estimates of net CO₂ exchange for the summer growing season (9 May?5 September 1995) indicate that nonacidic tundra was a net sink of ?31.7 ± 21.3 GgC (1 Gg = 10⁹ g), while shrub tundra lost 32.5 ± 6.3 GgC to the atmosphere (negative values denote net ecosystem CO₂ uptake). Acidic and wet sedge tundra were in balance, and when integrated for the entire Kuparuk River Basin (including aquatic surfaces), whole basin summer net CO₂ exchange was estimated to be in balance (?0.9 ± 50.3 GgC). Autumn to winter (6 September 1995–8 May 1996) estimates of net CO₂ flux indicate that acidic, nonacidic, and shrub tundra landforms were all large sources of CO₂ to the atmosphere (75.5 ± 8.3, 96.4 ± 11.4, and 43.3 ± 4.7 GgC for acidic, nonacidic, and shrub tundra, respectively). CO₂ loss from wet sedge surfaces was not substantially different from zero, but the large losses from the other terrestrial landforms resulted in a whole basin net CO₂ loss of 217.2 ± 24.1 GgC during the 1995–96 cold season. When integrated for the 1995–96 annual cycle, acidic (66.4 + 25.25 GgC), nonacidic (64.7 ± 29.2 GgC), and shrub tundra (75.8 ± 8.4 GgC) were substantial net sources of CO₂ to the atmosphere, while wet sedge tundra was in balance (0.4 + 0.8 GgC). The Kuparuk River Basin as a whole was estimated to be a net CO₂ source of 218.1 ± 60.6 GgC over the 1995–96 annual cycle. Compared to direct measurements of regional net CO₂ flux obtained from aircraft‐based eddy covariance, the scaling procedure provided realistic estimates of CO₂ exchange during the summer growing season. Although winter estimates could not be assessed directly using aircraft measurements of net CO₂ exchange, the estimates reported here are comparable to measured values reported in the literature. Thus, we have high confidence in the summer estimates of net CO₂ exchange and reasonable confidence in the winter net CO₂ flux estimates for terrestrial landforms of the Kuparuk river basin. Although there is larger uncertainty in the aquatic estimates, the small surface area of aquatic surfaces in the Kuparuk river basin (≈ 5%) presumably reduces the potential for this uncertainty to result in large errors in basin‐wide CO₂ flux estimates.     

Phylogeny of Chinese Allium (Liliaceae) using PCR-RFLP analysis

Eighteen representative species were selected from all the nine sections of Chinese Allium on the basis of the classification of morphology and cytotaxonomy. The trnK and rpL16 gene fragments of chloroplast DNA were amplified from 18 species by PCR method. The two cpDNA fragments were digested by 26 restriction enzymes, and 303 polymorphic restriction sites were found, of which 163 were informative. The restriction site data were analyzed with PAUP (version 3.1.1) and MEGA (version 1.01) as well as PHYLIP. As a result, the genus Allium could be classified into six subgenera. The recognition of Sect. Anguinum in the Flora of China is reasonable, Sect. Rhizirideum, Sect. Haplostemon and Sect. Cepa are not monophyletic. The infrageneric system of this genus was also discussed.     

微流控芯片细胞捕获分离方法概述

细胞捕获分离是免疫学、诊断检测、病理研究等学科经常用到的生物学实验方法.近年来,微流控芯片平台的细胞捕获分离方式花样繁多,层出不穷,它具有可快速检测、所需样本量少、节约试剂、成本低廉等优势.本文主要对近年来多种微流控细胞捕获分离的方法,以免疫捕获分离和无标签细胞分离两类对其进行介绍.免疫捕获分离是较为传统的细胞捕获分离方式,它的特异性好、捕获分离后的细胞纯度较高.无标签细胞分离是近几年热门发展的技术手段,它采用物理学与生物学相结合的方式,能较好地保持细胞的完整性和生物活性.细胞捕获分离在微流控平台的应用虽然发展迅速,但其在工业化生产和微型化整合等方面还存在一些问题,只有解决生产问题,细胞捕获分离在微流控平台的应用才真正具有实际价值,可以真正作为一种技术手段用于日常的实验操作和医学检测中.就目前而言,细胞捕获分离在微流控芯片中仍具有很大的发展前景.     

泗礁沙滩碎波带仔稚鱼群落结构的时空变化

2010年8月—2011年8月每月大潮期间,在泗礁沙滩8个站位点水深0.5—1.5 m处,沿海岸平行拖曳小型拖网(1m×4m,网目1 mm)采集仔稚鱼样本。共采集仔稚鱼1718尾(隶属于28科46种)。仔稚鱼群落的季节变化显示,根据种类生活史及其对沙滩碎波带利用模式的差异,主要优势种鲻Mugil cephalus、中国花鲈Lateolabrax maculatus、鳀Engraulis japonicus、相模虾虎鱼Sagamia geneionema、细鳞鯻Terapon jarbua、鮻Liza haematocheila及中华侧带小公鱼Stolephorus chinensis分别在3月、5月、6月与8月进入碎波带,栖息时间在1—4个月不等;月份聚类和排序将8个月份(渔获物稀少甚至无渔获月份舍弃)分为2011年3—5月与2010年8—9月、2011年6—8月2个群组。群落的空间结构显示,St.5与St.7分别具有最多的种类数和个体数,说明仔稚鱼偏好栖息于内湾浅滩;但各主要优势种对内湾沙滩的选择有一定的种类特异性,其具体原因有待进一步完调查完善与分析;站点聚类和排序将8个站位分为4组:Sts.1—Sts.2与Sts.7—Sts.8聚为1组,St.5与St.6聚为1组,St.3、St.4分别为1组。各主要优势种对碎波带的利用时间、方式及对沙滩生境的选择均具有一定的种类特异性,其对沙滩生境选择的影响因子有待进一步研究。     

Reversible electropermeabilisation of human and rat blood platelets: evaluation of morphological and functional integrity 'in vitro' and 'in vivo'

A high-voltage discharge procedure has been developed for permeabilising the plasma membranes of both human and rat blood platelets. The cells can be resealed by incubation at 37 degrees C, show less than 4% loss of lactate dehydrogenase (LDH) implying minimal cell lysis and also have well maintained morphological and functional integrity. The prototype apparatus used at field strengths between 6 and 8 kV/cm produces membrane pores which allow free diffusion of low molecular weight substances such as adenine nucleotides, inositol phosphate and fluorescent dyes. Two properties, namely Ca2+-induced secretion of granule stored 5-hydroxytryptamine (5HT) and inositol 1,4,5-trisphosphate (IP3)-induced release of intracellularly sequestered 45Ca, which are both well expressed immediately after permeabilisation, are essentially abolished after resealing. The efficiency of permeabilisation and resealing can be simply monitored by shifts in 'apparent platelet volume' using a resistive particle counter (Coulter). Permeabilised platelets show a shift in modal volumes from a control range 4-7 fl to 10-15 fl. Resealing restores these modal volumes to the original control range. Encapsulation of the fluorochrome, Lucifer yellow (Mr 550), during permeabilisation revealed that after resealing greater than 85% of rat platelets, and close to 100% human platelets, contained the encapsulated dye. The initial rates and % aggregation responses of both human and rat platelets to collagen, thrombin and the thromboxane A2-mimetic U46619 remained essentially normal after permeabilisation and resealing further illustrating the maintenance of functional competence following treatment. Resealed rat platelets reinfused into the circulation after labelling with [111In]indium oxine gave survival curves similar to those of control platelets. Therefore, this reversible permeabilisation procedure may allow the use of autologous or heterologous platelets as carrier vehicles for the delivery of drugs and other agents 'in vivo'.