海参微卫星DNA的筛选与克隆的初步分析

微卫星DNA(microsatelliteDNA)广泛存在于真核生物的基因组中。由于其具有突变频率快、多态性丰富、呈共显性遗传、通用性等特点,已成为近年来被广泛应用的分子遗传标记。对刺参基因组DNA的提取及其微卫星的筛选与克隆进行了初步分析。     

Versican G3 promotes mouse mammary tumor cell growth, migration, and metastasis by influencing EGF receptor signaling

Increased versican expression in breast tumors is predictive of relapse and has negative impact on survival rates. The C-terminal G3 domain of versican influences local and systemic tumor invasiveness in pre-clinical murine models. However, the mechanism(s) by which G3 influences breast tumor growth and metastasis is not well characterized. Here we evaluated the expression of versican in mouse mammary tumor cell lines observing that 4T1 cells expressed highest levels while 66c14 cells expressed low levels. We exogenously expressed a G3 construct in 66c14 cells and analyzed its effects on cell proliferation, migration, cell cycle progression, and EGFR signaling. Experiments in a syngeneic orthotopic animal model demonstrated that G3 promoted tumor growth and systemic metastasis in vivo. Activation of pERK correlated with high levels of G3 expression. In vitro, G3 enhanced breast cancer cell proliferation and migration by up-regulating EGFR signaling, and enhanced cell motility through chemotactic mechanisms to bone stromal cells, which was prevented by inhibitor AG 1478. G3 expressing cells demonstrated increased CDK2 and GSK-3β (S9P) expression, which were related to cell growth. The activity of G3 on mouse mammary tumor cell growth, migration and its effect on spontaneous metastasis to bone in an orthotopic model was modulated by up-regulating the EGFR-mediated signaling pathway. Taken together, EGFR-signaling appears to be an important pathway in versican G3-mediated breast cancer tumor invasiveness and metastasis.     

A Strategy for the Proliferation of Ulva prolifera,Main Causative Species of Green Tides,with Formation of Sporangia by Fragmentation

Ulva prolifera, a common green seaweed, is one of the causative species of green tides that occurred frequently along the shores of Qingdao in 2008 and had detrimental effects on the preparations for the 2008 Beijing Olympic Games sailing competition, since more than 30 percent of the area of the games was invaded. In view of the rapid accumulation of the vast biomass of floating U. prolifera in green tides, we investigated the formation of sporangia in disks of different diameters excised from U. prolifera, changes of the photosynthetic properties of cells during sporangia formation, and development of spores. The results suggested that disks less than 1.00 mm in diameter were optimal for the formation of sporangia, but there was a small amount of spore release in these. The highest percentage of area of spore release occurred in disks that were 2.50 mm in diameter. In contrast, sporangia were formed only at the cut edges of larger disks (3.00 mm, 3.50 mm, and 4.00 mm in diameter). Additionally, the majority of spores liberated from the disks appeared vigorous and developed successfully into new individuals. These results implied that fragments of the appropriate size from the U. prolifera thalli broken by a variety of factors via producing spores gave rise to the rapid proliferation of the seaweed under field conditions, which may be one of the most important factors to the rapid accumulation of the vast biomass of U. prolifera in the green tide that occurred in Qingdao, 2008.     

Immune protection induced on day 10 following administration of the 2009 A/H1N1 pandemic influenza vaccine

Background

The 2009 swine-origin influenza virus (S-OIV) H1N1 pandemic has caused more than 18,000 deaths worldwide. Vaccines against the 2009 A/H1N1 influenza virus are useful for preventing infection and controlling the pandemic. The kinetics of the immune response following vaccination with the 2009 A/H1N1 influenza vaccine need further investigation.

Methodology/Principal Findings

58 volunteers were vaccinated with a 2009 A/H1N1 pandemic influenza monovalent split-virus vaccine (15 µg, single-dose). The sera were collected before Day 0 (pre-vaccination) and on Days 3, 5, 10, 14, 21, 30, 45 and 60 post vaccination. Specific antibody responses induced by the vaccination were analyzed using hemagglutination inhibition (HI) assay and enzyme-linked immunosorbent assay (ELISA). After administration of the 2009 A/H1N1 influenza vaccine, specific and protective antibody response with a major subtype of IgG was sufficiently developed as early as Day 10 (seroprotection rate: 93%). This specific antibody response could maintain for at least 60 days without significant reduction. Antibody response induced by the 2009 A/H1N1 influenza vaccine could not render protection against seasonal H1N1 influenza (seroconversion rate: 3% on Day 21). However, volunteers with higher pre-existing seasonal influenza antibody levels (pre-vaccination HI titer ≥1∶40, Group 1) more easily developed a strong antibody protection effect against the 2009 A/H1N1 influenza vaccine as compared with those showing lower pre-existing seasonal influenza antibody levels (pre-vaccination HI titer <1∶40, Group 2). The titer of the specific antibody against the 2009 A/H1N1 influenza was much higher in Group 1 (geometric mean titer: 146 on Day 21) than that in Group 2 (geometric mean titer: 70 on Day 21).

Conclusions/Significance

Recipients could gain sufficient protection as early as 10 days after vaccine administration. The protection could last at least 60 days. Individuals with a stronger pre-existing seasonal influenza antibody response may have a relatively higher potential for developing a stronger humoral immune response after vaccination with the 2009 A/H1N1 pandemic influenza vaccine.     

Distinct Expression Levels and Patterns of Stem Cell Marker,Aldehyde Dehydrogenase Isoform 1 (ALDH1), in Human Epithelial Cancers

Aldehyde dehydrogenase isoform 1 (ALDH1) has been proved useful for the identification of cancer stem cells. However, our knowledge of the expression and activity of ALDH1 in common epithelial cancers and their corresponding normal tissues is still largely absent. Therefore, we characterized ALDH1 expression in 24 types of normal tissues and a large collection of epithelial tumor specimens (six cancer types, n = 792) by immunohistochemical staining. Using the ALDEFUOR assay, ALDH1 activity was also examined in 16 primary tumor specimens and 43 established epithelial cancer cell lines. In addition, an ovarian cancer transgenic mouse model and 7 murine ovarian cancer cell lines were analyzed. We found that the expression levels and patterns of ALDH1 in epithelial cancers are remarkably distinct, and they correlate with their corresponding normal tissues. ALDH1 protein expression levels are positively correlated with ALDH1 enzymatic activity measured by ALDEFLUOR assay. Long-term in vitro culture doesn''t significantly affect ALDH1 activity in epithelial tumor cells. Consistent with research on other cancers, we found that high ALDH1 expression is significantly associated with poor clinical outcomes in serous ovarian cancer patients (n = 439, p = 0.0036). Finally, ALDH^br tumor cells exhibit cancer stem cell properties and are resistant to chemotherapy. As a novel cancer stem cell marker, ALDH1 can be used for tumors whose corresponding normal tissues express ALDH1 in relatively restricted or limited levels such as breast, lung, ovarian or colon cancer.     

Development and characterization of West Nile virus replicon expressing secreted Gaussia Luciferase

We developed a Gaussia luciferase (Gluc) reporter replicon of West Nile virus (WNV) and used it to quantify viral translation and RNA replication. The advantage of the Gluc replicon is that Gaussia luciferase is secreted into the culture medium from cells transfected with Gluc replicon RNA, and the medium can be assayed directly for luciferase activity. Using a known Flavivirus inhibitor (NITD008), we demonstrated that the Gluc-WNV replicon could be used for antiviral screening. The Gluc-WNV-Rep will be useful for research in antiviral drug development programs, as well as for studying viral replication and pathogenesis of WNV.     

Seed dormancy release and germination characteristics of <i>Corispermum lehmannianum</i> Bunge,an endemic species in the Gurbantunggut desert of China

Seed dormancy release and germination of Corispermum lehmannianum Bunge were tested using various treatments: temperature, cold stratification, gibberelins (GA3), dry storage and sand burial. Results showed that temperature and light did not affect the germination of fresh seeds, cold stratification and GA3 could improve seed germination, whereas dry storage and sand burial did not. The germination percentage was highest at 35/20 °C after the cold stratification and GA3 treatments. Corispermum lehmannianum seeds were classified as non-deep, Type-2, physiological dormancy (PD), whose seed dormancy could be released by cold stratification and GA3.     

甘肃徐家城旧石器遗址石制品拼合研究

徐家城遗址是甘肃省新发现的含丰富旧石器文化遗物的遗址,主要埋藏于水洛河二级阶地上覆的马兰黄土中,年代在距今4.3~2.3万年间。徐家城遗址的拼合分析显示,4B、4C层剥片与石器加工行为是石制品空间分布形成的主要原因,但可能存在遗址使用时古人类对石制品位置改变的行为;4A层石制品零散分布可能反映了此时遗址处于古人类活动的边缘地带。遗址剥片方法以硬锤锤击法为主,石核剥片序列主要以转向为主,古人类对石料进行单层或多层开发;同时部分石器破碎后古人类可能对其进行再利用。平面分布显示发掘区内可能存在两个相对集中的剥片区域,并且古人类在同一区域剥片和加工石器。然因遗址石制品原料性质、发掘面积等局限性,拼合分析的优势并未在此项研究中尽现。今后此类研究应结合其他研究方法,综合多方面信息解释遗址形成过程、石器技术及遗址空间利用方式。     

全球陆地保护地60年增长情况分析和趋势预测

保护地建设已成为全球生物多样性保护的第一道防线。掌握其在不同尺度上的发展状况和变化趋势对保护地规划和建设有重要意义。针对已有研究在时间跨度、空间尺度以及结果对比方面的不足, 本文基于世界保护地数据库(World Database of Protected Areas, http://www.protectedplanet.net), 对全球、洲际、地区及国家尺度1950-2013年陆地保护地的增长情况进行描述和短期预测。结果发现: (1)全球保护地增长速率不断加快, 特别是在20世纪90年代以后。(2)洲际和地区保护地发展大致呈现3种增长趋势: 在美洲及大洋洲, 多数地区的保护地增长速率一直在加快; 在亚洲和欧洲, 多数地区的发展高峰出现在20世纪80、90年代; 在非洲, 多数地区的发展高峰为20世纪70年代及21世纪前10年。(3)各国保护地建设存在不平衡性, 仍有近一半国家的陆地保护地比例小于10%, 但这种差距随时间的推移有缩小的趋势。(4)绝大部分保护地增速均匀性低的国家分布在非洲。(5)虽然全球的《爱知生物多样性目标》在2020年预计不能完成, 但包括中国在内的22个国家有望如期达到目标。本文结果为未来保护地规划和建设工作的开展提供了参考依据。     

全球陆地保护地与城市距离变化分析

在城市用地和保护地都快速扩张的背景下, 保护地与城市之间的距离在急剧缩小, 然而这并未引起人们的足够关注。城市对保护地的负面影响具有尺度效应, 且随着距离的接近, 负面影响也将加剧, 因此保护地与城市距离的远近可成为衡量负面影响大小的重要依据。本文基于全球保护地和大城市及城市用地分布数据, 对全球、洲际、地区和国家尺度1950-2010年城市与保护地距离的变化进行分析。结果表明: (1)在4个尺度上, 保护地与城市的距离都在不断缩小。欧洲和西欧是保护地与城市距离最近的洲和地区, 而大洋洲和澳大利亚及新西兰则是距离最远的洲和地区。在面积排名前20的保护地大国中, 中国是保护地与城市平均最近距离最小的国家, 其与50万以上人口城市的平均最近距离在2010年仅为143.5 km。(2)根据城市与保护地距离的现状及其变化, 可将保护地排名前60的国家分成5类: (a)距离很近, 靠近速度慢, 如西欧国家; (b)距离近, 靠近速度适中, 如中国、美国; (c)距离较近, 靠近速度快, 如沙特阿拉伯、厄瓜多尔; (d)距离较远, 靠近速度较慢, 如巴西、加拿大、俄罗斯联邦; (e)距离远, 靠近速度较快, 如澳大利亚、非洲大多数国家。(3)全球范围内, 越来越多生物多样性较高的保护地将面临被城市影响的境地。该结果以期引起大家对全球保护地与城市距离急剧缩短现象的关注和警惕。