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   3167篇
  免费   237篇
  国内免费   279篇
  2024年   5篇
  2023年   51篇
  2022年   116篇
  2021年   202篇
  2020年   128篇
  2019年   155篇
  2018年   135篇
  2017年   124篇
  2016年   175篇
  2015年   225篇
  2014年   237篇
  2013年   281篇
  2012年   314篇
  2011年   277篇
  2010年   165篇
  2009年   131篇
  2008年   166篇
  2007年   140篇
  2006年   107篇
  2005年   83篇
  2004年   71篇
  2003年   57篇
  2002年   49篇
  2001年   32篇
  2000年   27篇
  1999年   32篇
  1998年   18篇
  1997年   21篇
  1996年   25篇
  1995年   16篇
  1994年   22篇
  1993年   12篇
  1992年   11篇
  1991年   11篇
  1990年   11篇
  1989年   8篇
  1988年   6篇
  1987年   3篇
  1986年   4篇
  1985年   6篇
  1984年   2篇
  1983年   3篇
  1981年   2篇
  1979年   3篇
  1978年   2篇
  1974年   1篇
  1973年   2篇
  1972年   1篇
  1971年   2篇
  1968年   3篇
排序方式: 共有3683条查询结果,搜索用时 593 毫秒
991.
Tang  Pei-jia  Zhang  Zhi-hui  Niu  Li-li  Gu  Cheng-bo  Zheng  Wen-yi  Cui  Hai-chao  Yuan  Xiao-han 《Biotechnology letters》2021,43(7):1371-1383

The study aimed to characterize a novel vitexin-producing endophytic fungus Fusarium solani G6 from Cajanus cajan, improve its capability for producing vitexin and evaluate its osteoblastic proliferation activity. A total of 153 endophytic fungi, classified into 6 genera, were isolated from C. cajan. Among them, only one strain, endophyte G6 identified as Fusarium solani, was found to produce vitexin. After the optimization of fermentation conditions, the highest vitexin yield (18.72 mg/L) for the strain was observed in PDB liquid medium containing 20.54 g/L of glucose and 8.90 g/L of ammonium sulfate, at an initial medium pH of 5.1 and at 28 °C for 6 days of cultivation. Moreover, the fungal vitexin exhibited notable osteoblastic proliferation stimulating activity. A novel vitexin-producing endophytic fungus F. solani G6 was characterized from C. cajan for the first time. The findings highlighted its potential use for large-scale production of vitexin and might have a promising use as therapeutic agent for osteoporosis.

  相似文献   
992.
Kang  Bin  Pecl  Gretta T.  Lin  Longshan  Sun  Peng  Zhang  Peidong  Li  Yuan  Zhao  Linlin  Peng  Xin  Yan  Yunrong  Shen  Chunyan  Niu  Wentao 《Reviews in Fish Biology and Fisheries》2021,31(3):599-629
Reviews in Fish Biology and Fisheries - Globally, climate change impacts on marine ecosystems are evident in physical, chemical, and biological processes, and are generally more extensive in faster...  相似文献   
993.
为了初步探究魔芋热激转录因子HSFB1基因及其启动子的功能,以白魔芋Amorphophallus albus为试材,利用同源克隆方法获得长度为1 365 bp的AaHSFB1基因序列。qRT-PCR结果表明:AaHSFB1基因对热胁迫较敏感,在根中的表达量表现出先升后降的趋势,并在热处理1 h时表达丰度最高;在热处理12 h时,叶片中的表达量也达到最高,在整个热处理时段内球茎中的表达量变化不大;亚细胞定位结果显示AaHSFB1定位于细胞核内。再利用FPNI-PCR法通过三轮步移扩增得到1 509 bp的AaHSFB1启动子序列。生物信息学分析表明:AaHSFB1含有热胁迫响应元件HSE及多种与植物发育及逆境应答相关的顺式作用元件。为进一步分析AaHSFB1启动子的功能,构建融合表达载体prAaHSFB1::GUS,利用农杆菌介导法转入拟南芥,热处理后对转基因拟南芥进行GUS组织化学染色鉴定,结果显示其表达部位主要在叶中。因此推测AaHSFB1可能在白魔芋抗外界逆境特别是热胁迫中起重要作用。  相似文献   
994.
【目的】随着合成生物学的发展,通过在细菌体内设计合成复杂、多功能的基因线路进行靶向治疗已经取得巨大进展。虽然这种使用细菌作为治疗传递系统,选择性地在体内释放有效治疗成分的方式具有极大优势,但是如何使细菌在代谢负荷增加较低的情况下有效地分泌功能蛋白并发挥作用依旧是一个难题。【方法】针对这一难题,本研究提供了一种新的策略,即以细菌中广泛存在的蛋白类杀菌素和丝状噬菌体等相关编码基因作为生物模块,通过对铜绿假单胞菌的这些内源生物模块的重新编排和组装,构建了一种能在特定条件下裂解并投放功能蛋白的工程菌。为了评价工程菌中构建的生物模块能否工作,本研究选择胞外多糖水解酶PelA和PslG作为工程菌投放的功能蛋白,以此构建了工程菌PAO1102。通过对铜绿假单胞菌生物被膜的破坏实验、抑制形成实验以及抗生素耐药性实验,检验PAO1102对铜绿假单胞菌生物被膜的破坏和预防效果。【结果】与对照组相比,工程菌PAO1102的处理可以显著破坏已形成的生物被膜并抑制生物被膜的形成,同时还可显著增强生物被膜中的细菌对妥布霉素的敏感性,且这些功能主要通过外界Pf4丝状噬菌体侵染并使工程菌裂解而释放功能蛋白这一途径实现的。【结论】本研究所构建的工程菌可以作为一种微生物工具,用于靶向破坏铜绿假单胞菌生物被膜。在后续的研究中可根据不同的需求,在工程菌中表达不同的功能基因并实现功能蛋白的定向投放,从而执行不同的生物学功能。  相似文献   
995.
Copper pollution poses a serious threat to the aquatic environment; however, in situ analytical methods for copper monitoring are still scarce. In the current study, Escherichia coli Rosetta was genetically modified to express OprF and ribB with promoter Pt7 and PcusC, respectively, which could synthesize porin and senses Cu2+ to produce riboflavin. The cell membrane permeability of this engineered strain was increased and its riboflavin production (1.45–3.56 μM) was positively correlated to Cu2+ (0–0.5 mM). The biosynthetic strain was then employed in microbial fuel cell (MFC) based biosensor. Under optimal operating parameters of pH 7.1 and 37°C, the maximum voltage (248, 295, 333, 352, and 407 mV) of the constructed MFC biosensor showed a linear correlation with Cu2+ concentration (0.1, 0.2, 0.3, 0.4, 0.5 mM, respectively; R2 = 0.977). The continuous mode testing demonstrated that the MFC biosensor specifically senses Cu2+ with calculated detection limit of 28 μM, which conforms to the common Cu2+ safety standard (32 μM). The results obtained with the developed biosensor system were consistent with the existing analytical methods such as colorimetry, flame atomic absorption spectrometry, and inductively coupled plasma optical emission spectrometry. In conclusion, this MFC‐based biosensor overcomes the signal conversion and transmission problems of conventional approaches, providing a fast and economic analytical alternative for in situ monitoring of Cu2+ in water.  相似文献   
996.
997.
HIV-1 functional cure requires sustained viral suppression without antiretroviral therapy. While effector-memory CD8+ T lymphocytes are essential for viremia control, few vaccines elicit such cellular immunity that could be potently recalled upon viral infection. Here, we investigated a program death-1 (PD1)-based vaccine by fusion of simian immunodeficiency virus capsid antigen to soluble PD1. Homologous vaccinations suppressed setpoint viremia to undetectable levels in vaccinated macaques following a high-dose intravenous challenge by the pathogenic SHIVSF162P3CN. Poly-functional effector-memory CD8+ T cells were not only induced after vaccination, but were also recalled upon viral challenge for viremia control as determined by CD8 depletion. Vaccine-induced effector memory CD8+ subsets displayed high cytotoxicity-related genes by single-cell analysis. Vaccinees with sustained viremia suppression for over two years responded to boost vaccination without viral rebound. These results demonstrated that PD1-based vaccine-induced effector-memory CD8+ T cells were recalled by AIDS virus infection, providing a potential immunotherapy for functional cure.  相似文献   
998.
Zhou  Chen  Huo  Yiping  Guo  Yiyuan  Niu  Qiqiang 《Plasmonics (Norwell, Mass.)》2021,16(5):1735-1743
Plasmonics - A metal-insulator-metal (MIM) waveguide consisting of two stub resonators and a ring resonator is proposed, which can be used as refractive index sensor and stop-band filter at the...  相似文献   
999.
Jiang  Teng  Diao  Xiaoli  Ding  Meili  Niu  Xiao  Wang  Chao  Qi  Yan  Jia  Wei  Pang  Lijuan  Hu  Wenhao  Zou  Hong  Li  Feng 《Journal of molecular histology》2021,52(3):539-544
Journal of Molecular Histology - Both clear cell renal carcinoma (ccRCC) and clear cell carcinoma of the ovary (CCOC) have a clear cytoplasmic morphological feature, hence it is difficult to...  相似文献   
1000.
Niu  Lijun  Chen  Lihong  Luo  Yanhua  Huang  Wenkao  Li  Yunsheng 《BMC anesthesiology》2021,21(1):1-6
This case demonstrates the severe electrolyte derangements that may present after a common therapy such as a bowel preparation for an outpatient procedure and the rare yet potential detrimental outcomes of those abnormalities. It also highlights the implications of long QT syndrome regarding pharmacology and treatment. We present a case of 48 year-old female with severe electrolyte derangements and long QT syndrome (LQTS) leading to Torsades de Pointes (TdP), pulseless ventricular fibrillation, and unsynchronized defibrillation in the post anesthesia care unit (PACU) after uneventful upper and lower endoscopy. This led to an unanticipated intensive care unit admission for aggressive electrolyte repletion, cardiology consultation, and implantable cardioverter defibrillator (ICD) placement. This is a rare presentation after an outpatient procedure that would have had a detrimental outcome if not promptly diagnosed and treated appropriately. Therefore, we aim to provide further insight into the diagnosis and treatment of severe hypokalemia and long QT syndrome resulting in Torsades de Pointes and ventricular fibrillation.  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