首页 | 本学科首页   官方微博 | 高级检索  
文章检索
  按 检索   检索词:      
出版年份:   被引次数:   他引次数: 提示:输入*表示无穷大
  收费全文   10846篇
  免费   929篇
  国内免费   905篇
  2024年   30篇
  2023年   138篇
  2022年   277篇
  2021年   480篇
  2020年   356篇
  2019年   441篇
  2018年   502篇
  2017年   345篇
  2016年   491篇
  2015年   694篇
  2014年   822篇
  2013年   900篇
  2012年   1034篇
  2011年   904篇
  2010年   593篇
  2009年   417篇
  2008年   556篇
  2007年   458篇
  2006年   406篇
  2005年   372篇
  2004年   357篇
  2003年   361篇
  2002年   281篇
  2001年   181篇
  2000年   148篇
  1999年   172篇
  1998年   95篇
  1997年   68篇
  1996年   55篇
  1995年   69篇
  1994年   61篇
  1993年   42篇
  1992年   59篇
  1991年   51篇
  1990年   34篇
  1989年   32篇
  1988年   36篇
  1987年   19篇
  1986年   27篇
  1985年   23篇
  1984年   25篇
  1983年   15篇
  1982年   14篇
  1981年   14篇
  1979年   12篇
  1974年   12篇
  1973年   21篇
  1972年   12篇
  1966年   12篇
  1958年   13篇
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
991.
Tan T  Tang X  Zhang J  Niu Y  Chen H  Li B  Wei Q  Ji W 《PloS one》2011,6(2):e17124
Trophoblast stem (TS) cells are ideal models to investigate trophectoderm differentiation and placental development. Herein, we describe the derivation of rabbit trophoblast stem cells from embryonic stem (ES) cells. Rabbit ES cells generated in our laboratory were induced to differentiate in the presence of BMP4 and TS-like cell colonies were isolated and expanded. These cells expressed the molecular markers of mouse TS cells, were able to invade, give rise to derivatives of TS cells, and chimerize placental tissues when injected into blastocysts. The rabbit TS-like cells maintained self-renewal in culture medium with serum but without growth factors or feeder cells, whilst their proliferation and identity were compromised by inhibitors of FGFs and TGF-β receptors. Taken together, our study demonstrated the derivation of rabbit TS cells and suggested the essential roles of FGF and TGF-β signalings in maintenance of rabbit TS cell self-renewal.  相似文献   
992.
Nie M  Wang Y  Yu J  Xiao M  Jiang L  Yang J  Fang C  Chen J  Li B 《PloS one》2011,6(3):e17961
Plant-microbe interactions are considered to be important processes determining the efficiency of phytoremediation of petroleum pollution, however relatively little is known about how these interactions are influenced by petroleum pollution. In this experimental study using a microcosm approach, we examined how plant ecophysiological traits, soil nutrients and microbial activities were influenced by petroleum pollution in Phragmites australis, a phytoremediating species. Generally, petroleum pollution reduced plant performance, especially at early stages of plant growth. Petroleum had negative effects on the net accumulation of inorganic nitrogen from its organic forms (net nitrogen mineralization (NNM)) most likely by decreasing the inorganic nitrogen available to the plants in petroleum-polluted soils. However, abundant dissolved organic nitrogen (DON) was found in petroleum-polluted soil. In order to overcome initial deficiency of inorganic nitrogen, plants by dint of high colonization of arbuscular mycorrhizal fungi might absorb some DON for their growth in petroleum-polluted soils. In addition, through using a real-time polymerase chain reaction method, we quantified hydrocarbon-degrading bacterial traits based on their catabolic genes (i.e. alkB (alkane monooxygenase), nah (naphthalene dioxygenase) and tol (xylene monooxygenase) genes). This enumeration of target genes suggests that different hydrocarbon-degrading bacteria experienced different dynamic changes during phytoremediation and a greater abundance of alkB was detected during vegetative growth stages. Because phytoremediation of different components of petroleum is performed by different hydrocarbon-degrading bacteria, plants' ability of phytoremediating different components might therefore vary during the plant life cycle. Phytoremediation might be most effective during the vegetative growth stages as greater abundances of hydrocarbon-degrading bacteria containing alkB and tol genes were observed at these stages. The information provided by this study enhances our understanding of the effects of petroleum pollution on plant-microbe interactions and the roles of these interactions in the phytoremediation of petroleum-polluted soil.  相似文献   
993.
Huo X  Qi X  Tang F  Zu R  Li L  Wu B  Qin Y  Ji H  Fu J  Wang S  Tian H  Hu Z  Yang H  Zhou M  Wang H  Zhu F 《PloS one》2011,6(3):e17995

Background

We investigated the seropositive rates and persistence of antibody against pandemic (H1N1) 2009 virus (pH1N1) in pregnant women and voluntary blood donors after the second wave of the pandemic in Nanjing, China.

Methodology/Principal Findings

Serum samples of unvaccinated pregnant women (n = 720) and voluntary blood donors (n = 320) were collected after the second wave of 2009 pandemic in Nanjing. All samples were tested against pH1N1 strain (A/California/7/2009) with hemagglutination inhibition assay. A significant decline in seropositive rates, from above 50% to about 20%, was observed in pregnant women and voluntary blood donors fifteen weeks after the second wave of the pandemic. A quarter of the samples were tested against a seasonal H1N1 strain (A/Brisbane/59/2007). The antibody titers against pH1N1 strain were found to correlate positively with those against seasonal H1N1 strain. The correlation was modest but statistically significant.

Conclusions and Significance

The high seropositive rates in both pregnant women and voluntary blood donors suggested that the pH1N1 virus had widely spread in these two populations. Immunity derived from natural infection seemed not to be persistent well.  相似文献   
994.
RNA/DNA sequences rich in guanine (G) can form a 4-strand structure, G-quadruplex, which has been extensively researched and observed in mammalian, fungi, and plants, with in vivo existence in eukaryotic cells. Compared with DNA quadruplex, the potential existence of RNA quadruplex appears to be generally rare; however, it is believed by some researchers to be more inevitable in vivo and speculated to play an important role where it exists. Recently, researches concerning the function of G-quadruplexes in RNAs commence, making much progress. However, there is no available review particularly focusing on RNA quadruplex till now as we know. Therefore, we decide to give a review to comprehensively summarize research progress on it. This review highlights the diverse topologies for RNA quadruplex structure and its effect factors; outlines the current knowledge of RNA quadruplex's physiological functions in biological systems, especially in gene expression; and presents the prospects of RNA quadruplex.  相似文献   
995.
目的:比较小羽贯众根状茎和叶提取物体外抑菌活性。方法:牛津杯法和96孔板法,通过测定抑菌圈大小、最小抑菌浓度(MIC)和最小杀菌浓度(MBC),比较小羽贯众根状茎和叶提取物分别对10种常见细菌的抑制作用,以及不同产地抑菌效果的比较。结果:小羽贯众的根状茎和叶的提取物对10种常见的细菌均表现出较强的抑菌活性,肺炎克雷伯氏菌对提取物最敏感。结论:小羽贯众提取物作为一种天然的抗菌物质,具有广谱抗菌效果,有较高的应用潜力和开发前景。  相似文献   
996.
目的:分析不同颧颞部骨折性质与颞部凹陷的相关性,评价颧颞部骨折术后并发颞部凹陷的防治效果。方法:对105例颧颞部骨折病例进行回顾性分析,52例患者行颞部凹陷修复术,采用头皮冠状切口,应用钛网修复颞部凹陷,术后通过长期随访评价治疗效果。结果:陈旧性骨折颞部凹陷的发生率显著高于新鲜骨折,但治疗后颞部外形均有明显改善。结论:钛网植入能有效地修复颧颞部骨折术后并发的颞部凹陷,但应把握手术时机及治疗方法。  相似文献   
997.
CBR2激活与小胶质细胞的活化和损伤的关系   总被引:1,自引:1,他引:0  
目的:探讨大麻素CBR2受体激动剂AM1241预处理对脂多糖(LPS)和γ-干扰素(IFN-γ)所致炎症反应对小胶质细胞活化和损伤的影响。方法:联用LPS和IFN-γ作为小胶质细胞损伤模型,将细胞分为Control组、AM1241组、LPS/IFN-γ组和AM1241+LPS/IFN-γ组;AM1241组和AM1241+LPS/IFN-γ组经AM1241预处理2h,LPS/IFN-γ组和AM1241+LPS/IFN-γ组用含LPS和IFN-γ的培养基培养24h。采用MTT法检测细胞代谢率,硝酸还原酶法检测细胞培养液中一氧化氮(NO)释放量,酶联免疫吸附剂测定细胞培养基中炎症因子释放量,倒置相差显微镜观察细胞形态。结果:与LPS/IFN-γ组相比,AM1241+LPS/IFN-γ组细胞代谢率明显升高(P<0.05),NO、TNF-α、IL-1β和IL-10释放量明显减少(P<0.05),活化和损伤程度明显减轻。结论:大麻素CBR2受体激动剂AM1241预处理可减轻LPS和IFN-γ对小胶质细胞的活化和损伤。  相似文献   
998.
目的:探讨四逆散对疲劳大鼠学习记忆及脑内GABA_B1受体的影响。方法:应用游泳训练和睡眠剥夺方法复制疲劳动物模型,运用Y-迷宫检测各组大鼠的学习记忆能力,采用免疫组化方法检测大鼠脑内GABA_B1受体阳性反应物的变化。结果:模型组与正常组相比,大鼠Y-迷宫正确反应率明显下降,错误反应率明显升高,大鼠脑内GABA_B1的阳性反应物数目明显增多;中药治疗组与模型组相比,正确反应率明显升高,错误反应次数明显下降,脑内GABA_B1受体的阳性反应物数目明显减少。结论:四逆散对疲劳大鼠学习记忆能力的下降具有改善作用。  相似文献   
999.
目的:探究云芝糖肽(PSP)对人急性淋巴母细胞白血病Molt-4细胞周期的影响。方法:采用流式细胞术BrdU/DNA双染法获得各时相细胞分布状况和细胞周期的动力学参数。结果:0.1 mg/mlPSP处理12 h后,G2/M期细胞百分比由对照组的11.09%减少至3.69%。DNA合成时间由12.10 h延长至108.40 h。24 h处理组中,S期细胞百分比由对照组的43.29%增加至67.26%,而G0/G1期和G2/M期细胞百分比均减少,G0/G1期细胞百分比由对照组的37.47%减少至27.43%,G2/M期细胞百分比由对照组的19.24%降低至5.31%。DNA合成时间更是由11.95 h延长至114.52 h。结论:PSP对人急性淋巴母细胞白血病Molt-4细胞周期的阻滞作用在于S期,该作用与DNA合成抑制有关。  相似文献   
1000.
Li S  Wang Y  Zhang J  Yang WH  Dai ZH  Zhu W  Yu XQ 《Molecular bioSystems》2011,7(4):1254-1262
Polyethylenimine (PEI, especially with M(w) of 25,000) has been known as an efficient gene carrier and a gold standard of gene transfection due to its high transfection efficiency (TE). However, high concomitant cytotoxicity limited the application of PEI. In this report, several cationic polymers derived from low molecular weight (LMW) PEI (M(w) 600) linked with diglycidyl adipate (DA-PEI) or its analogs (diglycidyl succinate, DS-PEI and diglycidyl oxalate, DO-PEI; D-PEIs for all 3 polymers) were prepared and characterized. GPC gave M(w)s of DA-PEI, DS-PEI and DO-PEI as 6861, 16,015 and 35,281, respectively. Moreover, degradation of the ester-containing DS-PEI was also confirmed by GPC. In addition, hydroxyls in these polymers could improve their water solubility. These polymers exhibited good ability to condense plasmid DNA into nanoparticles with the size of 120-250 nm. ζ-potentials of the polyplexes were found to be around +10-20 mV under weight ratios (polymer/DNA) from 0.5 to 32. Agarose gel retardation showed that DNA could be released from the polyplexes after being pre-incubated for 30 h. In vitro experiments were carried out and it was found that DS-PEI showed about 5 times of TE compared to that of the PEI/DNA polyplex under a weight ratio of 1 in A549 cells. Meanwhile, the cytotoxicity of D-PEIs assayed by MTT is lower than that of 25 kDa PEI in HEK293 cells. These results suggested that this series of PEI derivatives would be promising non-viral biodegradable vectors for gene delivery.  相似文献   
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号