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   2155篇
  免费   201篇
  2023年   8篇
  2022年   24篇
  2021年   43篇
  2020年   30篇
  2019年   62篇
  2018年   74篇
  2017年   66篇
  2016年   75篇
  2015年   120篇
  2014年   140篇
  2013年   170篇
  2012年   160篇
  2011年   204篇
  2010年   122篇
  2009年   69篇
  2008年   132篇
  2007年   121篇
  2006年   145篇
  2005年   121篇
  2004年   90篇
  2003年   93篇
  2002年   64篇
  2001年   18篇
  2000年   10篇
  1999年   13篇
  1998年   7篇
  1997年   9篇
  1996年   11篇
  1995年   8篇
  1994年   4篇
  1993年   6篇
  1992年   8篇
  1990年   8篇
  1989年   4篇
  1988年   7篇
  1987年   4篇
  1986年   7篇
  1985年   6篇
  1984年   6篇
  1983年   8篇
  1982年   5篇
  1981年   4篇
  1980年   5篇
  1978年   5篇
  1977年   8篇
  1976年   6篇
  1975年   15篇
  1974年   4篇
  1973年   6篇
  1970年   5篇
排序方式: 共有2356条查询结果,搜索用时 828 毫秒
81.
82.
83.
Regulation of autophagy in bovine mammary epithelial cells   总被引:1,自引:0,他引:1  
The bovine mammary gland undergoes intensive remodelling during the lactation cycle, and the escalation of this process is observed during dry periods. The main type of cell death responsible for bovine mammary gland involution is apoptosis; however, there are also a lot of cells exhibiting morphological features of autophagy during drying off. Our in vitro and in vivo studies of bovine mammary gland physiology suggest that the enhanced process of autophagy, observed at the end of lactation and during dry periods, is the result of: (1) decreased level of lactogenic hormones (GH, IGF-I), (2) decreased GH-R and IGF-IR alpha expression, (3) increased expression of auto/paracrine apoptogenic peptides (IGFBPs, TGFbeta), (4) increased influence of sex steroids (17beta-estradiol and progesterone) and (5) enhanced competition between the between the intensively developing fetus and the mother organism for nutritional and bioactive compounds. The above conditions may create a state of temporary malnutrition of mammary epithelial cells, which forces the cells to the induction of autophagy, as a mechanism for stabilizing intracellular supplies of energy and amino acids, especially during the enhanced activity of apoptogenic factors.  相似文献   
84.
The paper concerns the problem of fitting mathematical models of cell signaling pathways. Such models frequently take the form of sets of nonlinear ordinary differential equations. While the model is continuous in time, the performance index used in the fitting procedure involves measurements taken at discrete time moments. Adjoint sensitivity analysis is a tool which can be used for finding the gradient of a performance index in the space of parameters of the model. In the paper, a structural formulation of adjoint sensitivity analysis called the generalized backpropagation through time (GBPTT) is used. The method is especially suited for hybrid, continuous-discrete time systems. As an example, we use the mathematical model NF-kappaB of the regulatory module, which plays a major role in the innate immune response in animals.  相似文献   
85.
86.
Biological Trace Element Research - We analyzed cobalt (Co), chromium (Cr), and lead (Pb) concentrations in human semen and catalase CAT activity in seminal plasma and the effects of their...  相似文献   
87.

Background

During standard gene cloning, the recombinant protein appearing in bacteria as the result of expression leakage very often inhibits cell proliferation leading to blocking of the cloning procedure. Although different approaches can reduce transgene basal expression, the recombinant proteins, which even in trace amounts inhibit bacterial growth, can completely prevent the cloning process.

Methods

Working to solve the problem of DNase II-like cDNA cloning, we developed a novel cloning approach. The method is based on separate cloning of the 5′ and 3′ fragments of target cDNA into a vector in such a way that the short Multiple Cloning Site insertion remaining between both fragments changes the reading frame and prevents translation of mRNA arising as a result of promoter leakage. Subsequently, to get the vector with full, uninterrupted Open Reading Frame, the Multiple Cloning Site insertion is removed by in vitro restriction/ligation reactions, utilizing the unique restriction site present in native cDNA.

Results

Using this designed method, we cloned a coding sequence of AcDNase II that is extremely toxic for bacteria cells. Then, we demonstrated the usefulness of the construct prepared in this way for overexpression of AcDNase II in eukaryotic cells.

Conclusions

The designed method allows cloning of toxic protein coding sequences that cannot be cloned by standard methods.

General significance

Cloning of cDNAs encoding toxic proteins is still a troublesome problem that hinders the progress of numerous studies. The method described here is a convenient solution to cloning problems that are common in research on toxic proteins.  相似文献   
88.
89.
90.
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