全文获取类型
收费全文 | 1504篇 |
免费 | 70篇 |
国内免费 | 5篇 |
出版年
2022年 | 5篇 |
2021年 | 13篇 |
2019年 | 16篇 |
2018年 | 15篇 |
2017年 | 9篇 |
2016年 | 24篇 |
2015年 | 42篇 |
2014年 | 46篇 |
2013年 | 136篇 |
2012年 | 67篇 |
2011年 | 67篇 |
2010年 | 35篇 |
2009年 | 60篇 |
2008年 | 71篇 |
2007年 | 67篇 |
2006年 | 58篇 |
2005年 | 85篇 |
2004年 | 88篇 |
2003年 | 78篇 |
2002年 | 85篇 |
2001年 | 29篇 |
2000年 | 27篇 |
1999年 | 38篇 |
1998年 | 22篇 |
1997年 | 20篇 |
1996年 | 7篇 |
1995年 | 12篇 |
1994年 | 12篇 |
1993年 | 19篇 |
1992年 | 30篇 |
1991年 | 25篇 |
1990年 | 25篇 |
1989年 | 17篇 |
1988年 | 22篇 |
1987年 | 19篇 |
1986年 | 21篇 |
1985年 | 16篇 |
1984年 | 19篇 |
1983年 | 17篇 |
1982年 | 12篇 |
1981年 | 17篇 |
1980年 | 12篇 |
1979年 | 10篇 |
1978年 | 10篇 |
1977年 | 8篇 |
1976年 | 9篇 |
1975年 | 5篇 |
1974年 | 6篇 |
1973年 | 4篇 |
1972年 | 6篇 |
排序方式: 共有1579条查询结果,搜索用时 15 毫秒
1.
Immunological memory for T and B cells was studied in an in vitro culture system with spleen cells from mice primed with bovine serum albumin (BSA). Spleen cells taken from mice immunized at various times previously with a single intravenous injection of alum-precipitated (AP) BSA and bacterial endotoxin (ET) were cultured in Marbrook's system with dinitrophenylated (DNP) BSA as the in vitro antigen. In the cultures of spleen cells obtained from mice primed more than 14 days previously, an IgG-predominant anti-BSA response was generated. However, no anti-BSA response was observed in the culture of spleen cells taken from mice primed 7 days previously (day 7 spleen cells). The failure of day 7 spleen cells to generate an antibody response in vitro was shown to be attributable to both the lack of B memory cells and the effect of “suppressive” macrophages induced by ET. On the other hand, anti-BSA memory in the spleen of mice primed with AP-BSA plus ET and 2 months later challenged with AP-BSA matured within 7 days and declined rather quickly by 30 days after the challenge. The difference in the time course of the generation of memory between the spleen cells from primary and from secondary immunized mice might be attributable to the difference in the maturation of memory B cells, since the time course of the development of memory T cells after the secondary immunization was similar to that observed after primary immunization. 相似文献
2.
Yusuke Nakamura Michio Ogawa Takahiro Nishide Mitsuru Emi Goro Kosaki Seiichi Himeno Kenichi Matsubara 《Gene》1984,28(2):263-270
The nucleotide sequences of the cloned human salivary and pancreatic α-amylase cDNAs correspond to the continuous mRNA sequences of 1768 and 1566 nucleotides, respectively. These include all of the amino acid coding regions. Salivary cDNA contains 200 bp in the 5′-noncoding region and 32 in the 3′-noncoding region. Pancreatic cDNA contains 3 and 27 bp of 5′- and 3′-noncoding regions, respectively. The nucleotide sequence humology of the two cDNAs is 96% in the coding region, and the predicted amino acid sequences are 94% homologous.Comparison of the sequences of human α-amylase cDNAs with those previously obtained for mouse α-amylase genes (Hagenbuchle et al., 1980; Schibler et al., 1982) showed the possibility of gene conversion between the two genes of human α-amylase. 相似文献
3.
Demonstration of up-regulated IL 2 receptor expression on an in vitro cloned BCL1 subline 总被引:1,自引:0,他引:1
K Nakanishi T Hashimoto K Hiroishi K Matsui T Yoshimoto H C Morse J Furuyama T Hamaoka K Higashino W E Paul 《Journal of immunology (Baltimore, Md. : 1950)》1987,138(6):1817-1825
We have established BCL1 CL-3 cells capable of responding to B15-TRF and interleukin 2 (IL 2). This clone has both high affinity and low affinity receptors for IL 2 (IL 2R), but IL 2 by itself did not stimulate either proliferation or immunoglobulin (Ig) secretion. B15-TRF, which possesses both growth and differentiation activity, causes an increase in size of CL-3 cells and renders CL-3 cells responsive to IL 2, including an increased expression of IL 2R (eight-fold to 10-fold) and the differentiation of CL-3 cells into Ig secretion (60 to 80% of cultured cells). CL-3 cells pretreated with B15-TRF for 12 hr become competent to respond to IL 2 by up-regulation of IL 2R within 12 hr. In contrast CL-3 cells pretreated with IL 2 for 12 hr required 24 hr B15-TRF stimulation to result in IL 2R up-regulation. Thus the ordered action of B15-TRF and IL 2 is the most effective operational pathway for the up-regulation of IL 2R. This IL 2-mediated IL 2R up-regulation and induction of Ig synthesis depends upon the concentration of IL 2 in the culture. Both responses seem to be caused by IL 2 molecules bound to high affinity IL 2R. However, the possibility of involvement of low affinity IL 2R can not be vigorously excluded. In fact the level of IL 2 required for a response is far higher than that needed for activated T cell proliferation. This cloned BCL1 subline promises to be a useful tool for studying the regulation and mechanisms of B cell responses. 相似文献
4.
Y Inada K Ohwada T Yoshimoto S Kojima K Takahashi Y Kodera A Matsushima Y Saito 《Biochemical and biophysical research communications》1987,148(1):392-396
The activated magnetic modifier was synthesized from magnetite, alpha, omega-dicarboxymethylpoly(oxyethylene) and N-hydroxysuccinimide (Biochem. Biophys. Res. Commun., 145, 908-914, 1987). Urokinase was directly coupled with the activated magnetic modifier to obtain magnetic urokinase. The magnetic urokinase dispersed in saline and exerted high fibrinolytic activity (13.8 X 10(4) IU/mg protein), and was readily recovered from saline by magnetic force of 250 Oe. By applying magnetic force, the urokinase was attracted at our will and local fibrinolysis was achieved on fibrin gel in a petri dish. 相似文献
5.
Detection of point mutation in the tyrosinase gene of a Japanese albino patient by a direct sequencing of amplified DNA 总被引:7,自引:1,他引:6
Hideaki Kikuchi Satoshi Hara Seiichi Ishiguro Makoto Tamai Minro Watanabe 《Human genetics》1990,85(1):123-124
Summary Enzymatic DNA amplification and direct DNA sequencing were used to detect a mutation in the tyrosinase gene of an albino patient. Single-base change could be detected by direct sequencing. This base change (G to A) is thought to result in an amino acid change (Arg to Gln) in tyrosinase of the patient. 相似文献
6.
Investigations were made of the growth ofNelumbo nucifera, an aquatic higher plant, in a natural stand in Lake Kasumigaura. A rise of 1.0 m in the water level after a typhoon in August
1986 caused a subsequent decrease in biomass ofN. nucifera from the maximum of 291 g d.w. m−2 in July to a minimum of 75 g d.w. m−2. The biomass recovered thereafter in shallower regions. The underground biomass in October tended to increase toward the
shore. The total leaf area index (LAI) is the sum of LAI of floating leaves and emergent leaves. The maximum total LAI was
1.3 and 2.8 m2 m−2 in 1986 and 1987, respectively. LAI of floating leaves did not exceed 1 m2 m−2. The elongation rates of the petiole of floating and emergent leaves just after unrolling were 2.6 and 3.4 cm day−1, respectively. The sudden rise in water level (25 cm day−1) after the typhoon in August 1986 caused drowning and subsequent decomposition of the mature leaves. Only the young leaves
were able to elongate, allowing their laminae to reach the water surface. The fluctuation in water level, characterized by
the amplitude and duration of flooding and the time of flooding in the life cycle, is an important factor determining the
growth and survival ofN. nucifera in Lake Kasumigaura. 相似文献
7.
8.
Summary An immunohistochemical study of the production of the intermediate filaments [vimentin, cytokeratin, and glial filament acidic protein (GFAP)] during development of the pituitary gland was made by use of fetal and adult human pituitary tissue. Among these intermediate filament proteins in the anterior and intermediate lobes of the pituitary, cytokeratin is the first to appear, followed by GFAP and vimentin. However, only cytokeratin is seen during the period of morphogenesis of the pituitary gland, with the type-II subfamily cytokeratin 8 being the earliest to appear. Among the simple-epithelial-type cytokeratins, cytokeratins 8 and 19 were observed within the pituitary primordium during morphogenesis. Cells immunoreactive for cytokeratins 8 and 19 showed a heterogeneous three-dimensional distribution pattern in Rathke's pouch. Both cytokeratins 8 and 19 tended to be strongly positive at sites in the pituitary primordium where cells had become more loosely arranged (i.e., areas far from the diencephalon) but were only weakly positive in areas in which the epithelial cells were densely packed (i.e., areas closely associated with the diencephalon). It is concluded that, during the period of morphogenesis, Rathke's pouch has the intermediate filaments characteristic of simple epithelium and shows different immunoreactivity for simple-epithelial-type cytokeratins from place to place according to the extent of cellular differentiation. 相似文献
9.
Structural and functional analysis of a polyoma-related mammalian plasmid (L factor): the enhancer activity and plasmid establishment. 总被引:1,自引:1,他引:0 下载免费PDF全文
H Yoshimura Y Ikeda M Yoshimoto S Tamaki K Hanada T Kusano T Kohda H Saito M Oishi 《Nucleic acids research》1991,19(13):3633-3639
L factor is a unique plasmid DNA which was originally discovered in a subclone (B822) of mouse L cells at a high copy number (more than 5,000 copies/cell). The presence of L factor caused no detectable abnormalities to the plasmid-bearing cells. We determined the total DNA sequence of the L factor I (and a part of L factor II) and compared it with that of polyoma DNA. Both DNA are common to the general construction of DNA frames such as early, late and noncoding regions, suggesting the two to be closely related. On the other hand, the L factor DNA sequences differ substantially from that of polyoma in the DNA sequences corresponding to the polyoma large T antigen, capsid proteins and a portion of the enhancer region. In order to investigate the mechanism of plasmid establishment of L factor, we compared the enhancer activity, capacity of DNA replication and efficiency of plasmid establishment of L factor with those of polyoma. The results indicate that L factor enhancer activity and DNA replication capacity were considerably lower than those of polyoma, suggesting that these altered (lowered) activities associated with L factor contribute to the plasmidal establishment and stable maintenance of L factor. 相似文献
10.
Studies on prolyl endopeptidase from shakashimeji (Lyophyllum cinerascens): purification and enzymatic properties 总被引:1,自引:0,他引:1
High prolyl endopeptidase (post-proline cleaving enzyme) [EC 3.4.21.26] activity was detected in fruit bodies of shakashimeji (Lyophyllum cinerascens), tsukuritake (mushroom: Agaricus bisporus), hirohachichitake (Lactarius hygrophoroides), and yaburebenitake (Russula lepida) which belong to the genus Basidiomycetes. Cell-free extract of shakashimeji showed high activities of proline iminopeptidase and arylamidase as well as prolyl endopeptidase. The prolyl endopeptidase was purified from the extract of shakashimeji by sequential chromatographies on DEAE-Toyopearl, DEAE-Sephadex and hydroxyapatite, and high-performance liquid chromatography with a DEAE-5PW column. The purified enzyme was homogeneous as judged by disc gel electrophoresis. The enzyme was most active at pH 6.8 as checked with Z-Gly-Pro-beta-naphthylamide as a substrate and was stable in the range of pH 5.8-7.4. The isoelectric point of the enzyme was 5.2 and the molecular weight was estimated to be 76,000 by gel filtration on Sephadex G-150 and by sodium dodecyl sulfate (SDS) gel electrophoresis, suggesting that the enzyme was a monomer. The enzyme was completely inhibited by diisopropyl fluorophosphate (DFP), Z-Gly-Pro-CH2Cl, and Z-Pro-prolinal, while it was not inhibited by p-chloromercuribenzoate (PCMB), phenylmethylsulfonyl fluoride (PMSF), or metal chelators. It was estimated that at least five subsites were concerned with the enzyme-substrate binding. Among them, the S1, S2, and S1' sites showed high stereospecificity, as in mammalian, microbial, and plant enzymes. The enzyme hydrolyzed TRH at the carboxyl side of the proline residue. The mushroom enzyme, that was sensitive to DFP, Z-Pro-prolinal, and Z-Gly-Pro-CH2Cl, but not to PCMB, were quite similar in characteristics to the Flavobacterium enzyme.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献