全文获取类型
收费全文 | 5525篇 |
免费 | 414篇 |
国内免费 | 3篇 |
出版年
2023年 | 15篇 |
2022年 | 16篇 |
2021年 | 59篇 |
2020年 | 40篇 |
2019年 | 54篇 |
2018年 | 92篇 |
2017年 | 89篇 |
2016年 | 106篇 |
2015年 | 196篇 |
2014年 | 242篇 |
2013年 | 340篇 |
2012年 | 334篇 |
2011年 | 386篇 |
2010年 | 227篇 |
2009年 | 183篇 |
2008年 | 334篇 |
2007年 | 369篇 |
2006年 | 354篇 |
2005年 | 351篇 |
2004年 | 322篇 |
2003年 | 316篇 |
2002年 | 286篇 |
2001年 | 107篇 |
2000年 | 82篇 |
1999年 | 99篇 |
1998年 | 82篇 |
1997年 | 53篇 |
1996年 | 71篇 |
1995年 | 67篇 |
1994年 | 48篇 |
1993年 | 48篇 |
1992年 | 68篇 |
1991年 | 40篇 |
1990年 | 49篇 |
1989年 | 51篇 |
1988年 | 30篇 |
1987年 | 36篇 |
1986年 | 27篇 |
1985年 | 30篇 |
1984年 | 37篇 |
1983年 | 23篇 |
1982年 | 33篇 |
1981年 | 18篇 |
1980年 | 24篇 |
1979年 | 22篇 |
1978年 | 15篇 |
1977年 | 17篇 |
1975年 | 7篇 |
1973年 | 7篇 |
1971年 | 6篇 |
排序方式: 共有5942条查询结果,搜索用时 22 毫秒
1.
Abstract Clostridium perfringens alpha-toxin was produced in a protein-hyperproducing strain, Bacillus brevis 47, by cloning the gene into the constructed expression-secretion vector which has the multiple promoters and the signal peptide coding region of an outer cell wall protein gene. The amount of alpha-toxin produced by the B. brevis 47 transformant carrying the gene was approximately 10 times greater than that produced by a B. subtilis transformant carrying the toxin gene. Biological activities and the N-terminal amino acid sequence of the toxin secreted by the B. brevis 47 transformant were identical to those of wild-type alpha-toxin. 相似文献
2.
Yoshitake Takada 《Ecological Research》1996,11(3):371-379
Monodonta labio (Gastropoda: Trochidae) occurs in a wide tidal zone on a boulder-covered shore in Amakusa, Japan. To investigate sources
of variation in reproductive output within a population, the fecundity ofM. labio was estimated. Regressions of gonad egg counts on shell width were calculated from samples collected bi-weekly at three tidal
zones: high, mid and low intertidal. Seasonal fluctuations in the regression revealed that individual females spawn at least
three times a year. Variation in fecundity between the three tidal zones was not detected at any time in standard 12 mm snails.
Due to the high growth rate in the low zone during the reproductive season, annual fecundity in the low zone was larger than
that in the high and mid zones. Thus, tidal zone variation in fecundity ofM. labio was a result of growth variation between tidal zones. 相似文献
3.
Takahiro Watanabe Yohei Narita Masahiro Yoshida Yoshitaka Sato Fumi Goshima Hiroshi Kimura Takayuki Murata 《Journal of virology》2015,89(19):10120-10124
Epstein-Barr virus (EBV) is a gammaherpesvirus, associated with infectious mononucleosis and various types of malignancy. We focused here on the BDLF4 gene of EBV and identified it as a lytic gene, expressed with early kinetics. Viral late gene expression of the BDLF4 knockout strain was severely restricted; this could be restored by an exogenous supply of BDLF4. These results indicate that BDLF4 is important for the EBV lytic replication cycle, especially in late gene expression. 相似文献
4.
5.
The oxygen cost of transport per unit distance (CoT; mL·kg-1·km-1) shows a U-shaped curve as a function of walking speed (v), which includes a particular walking speed minimizing the CoT, so called economical speed (ES). The CoT-v relationship in running is approximately linear. These distinctive walking and running CoT-v relationships give an intersection between U-shaped and linear CoT relationships, termed the energetically optimal transition speed (EOTS). This study investigated the effects of subtracting the standing oxygen cost for calculating the CoT and its relevant effects on the ES and EOTS at the level and gradient slopes (±5%) in eleven male trained athletes. The percent effects of subtracting the standing oxygen cost (4.8 ± 0.4 mL·kg-1·min-1) on the CoT were significantly greater as the walking speed was slower, but it was not significant at faster running speeds over 9.4 km·h-1. The percent effect was significantly dependent on the gradient (downhill > level > uphill, P < 0.001). The net ES (level 4.09 ± 0.31, uphill 4.22 ± 0.37, and downhill 4.16 ± 0.44 km·h-1) was approximately 20% slower than the gross ES (level 5.15 ± 0.18, uphill 5.27 ± 0.20, and downhill 5.37 ± 0.22 km·h-1, P < 0.001). Both net and gross ES were not significantly dependent on the gradient. In contrast, the gross EOTS was slower than the net EOTS at the level (7.49 ± 0.32 vs. 7.63 ± 0.36 km·h-1, P = 0.003) and downhill gradients (7.78 ± 0.33 vs. 8.01 ± 0.41 km·h-1, P < 0.001), but not at the uphill gradient (7.55 ± 0.37 vs. 7.63 ± 0.51 km·h-1, P = 0.080). Note that those percent differences were less than 2.9%. Given these results, a subtraction of the standing oxygen cost should be carefully considered depending on the purpose of each study. 相似文献
6.
Changes in Targeting Efficiencies of Proteins to Plant Microbodies Caused by Amino Acid Substitutions in the Carboxy-terminal Tripeptide 总被引:7,自引:0,他引:7
Hayashi Makoto; Aoki Masahiro; Kondo Maki; Nishimura Mikio 《Plant & cell physiology》1997,38(6):759-768
It has been demonstrated that the carboxyl terminus of microbodyenzymes functions as a targeting signal to microbodies in higherplants. We have examined an ability of 24 carboxy-terminal aminoacid sequences to facilitate the transport of a cytosolic passengerprotein, ß-glucuroni-dase, into microbodies in greencotyledonary cells of trans-genic Arabidopsis. Immunoelectronmicroscopic analysis revealed that carboxy-terminal tripeptidesequences of the form [C/A/S/P]-[K/R]-[I/L/M] function as amicrobody-targeting signal, although tripeptides with prolineat the first amino acid position and isoleucine at the carboxylterminus show weak targeting efficiencies. All known micro-bodyenzymes that are synthesized in a form similar in size to themature molecule, except catalase, contain one of these tripeptidesequences at their carboxyl terminus. (Received April 14, 1997; Accepted April 8, 1997) 相似文献
7.
Nobukazu Suganuma Satoru Ito Hiromichi Aso Masashi Kondo Mitsuo Sato Masahiro Sokabe Yoshinori Hasegawa 《PloS one》2012,7(9)
It is suggested that migration of airway smooth muscle (ASM) cells plays an important role in the pathogenesis of airway remodeling in asthma. Increases in intracellular Ca2+ concentrations ([Ca2+]i) regulate most ASM cell functions related to asthma, such as contraction and proliferation. Recently, STIM1 was identified as a sarcoplasmic reticulum (SR) Ca2+ sensor that activates Orai1, the Ca2+ channel responsible for store-operated Ca2+ entry (SOCE). We investigated the role of STIM1 in [Ca2+]i and cell migration induced by platelet-derived growth factor (PDGF)-BB in human ASM cells. Cell migration was assessed by a chemotaxis chamber assay. Human ASM cells express STIM1, STIM2, and Orai1 mRNAs. SOCE activated by thapsigargin, an inhibitor of SR Ca2+-ATPase, was significantly blocked by STIM1 siRNA and Orai1 siRNA but not by STIM2 siRNA. PDGF-BB induced a transient increase in [Ca2+]i followed by sustained [Ca2+]i elevation. Sustained increases in [Ca2+]i due to PDGF-BB were significantly inhibited by a Ca2+ chelating agent EGTA or by siRNA for STIM1 or Orai1. The numbers of migrating cells were significantly increased by PDGF-BB treatment for 6 h. Knockdown of STIM1 and Orai1 by siRNA transfection inhibited PDGF-induced cell migration. Similarly, EGTA significantly inhibited PDGF-induced cell migration. In contrast, transfection with siRNA for STIM2 did not inhibit the sustained elevation of [Ca2+]i or cell migration induced by PDGF-BB. These results demonstrate that STIM1 and Orai1 are essential for PDGF-induced cell migration and Ca2+ influx in human ASM cells. STIM1 could be an important molecule responsible for airway remodeling. 相似文献
8.
Haruhiko Takada Yoshiyuki Hirachi Hideo Hashizume Shozo Kotani 《Microbiology and immunology》1980,24(11):1079-1090
Cytoplasmic membranes of L-forms of Staphylococcus aureus exerted a strong mitogenic effect on splenocytes of athymic nude mice as well as normal mice, while a cytoplasmic fraction of the same bacteria did not show definite mitogenicity. The mitogenic principle(s) of the membrane fraction was resistant to treatment with trypsin and was heat stable (at 100 C for 10 min). The active principle(s) in the insoluble residue of the membrane fraction digested with trypsin was not extracted with cold acetone, but could be solubilized by extraction with a cold chloroform-methanol mixture (2:1, v/v). The mitogenic principle(s) in the extract was fractionated by silicic acid column chromatography. Among five fractions separated by chromatography, fractions eluted with chloroform-methanol mixtures (1:1 and 1:20, v/v) were found to be strongly mitogenic. The cytoplasmic membranes of the L-forms also exerted a definite mitogenic effect on guinea pig splenocytes, but not on the thymocytes. 相似文献
9.
Yutaro Yamagata Yukiko Muramoto Sho Miyamoto Keiko Shindo Masahiro Nakano Takeshi Noda 《Microbiology and immunology》2019,63(5):164-171
Defective interfering (DI) influenza viruses carry a large deletion in a gene segment that interferes with the replication of infectious virus; thus, such viruses have potential for antiviral therapy. However, because DI viruses cannot replicate autonomously without the aid of an infectious helper virus, clonal DI virus stocks that are not contaminated with helper virus have not yet been generated. To overcome this problem, we used reverse genetics to generate a clonal DI virus with a PB2 DI gene, amplified the clonal DI virus using a cell line stably expressing the PB2 protein, and confirmed its ability to interfere with infectious virus replication in vitro. Thus, our approach is suitable for obtaining purely clonal DI viruses, will contribute to the understanding of DI virus interference mechanisms and can be used to develop DI virus‐based antivirals. 相似文献
10.
Understanding how defects in mechanotransduction affect cell‐to‐cell variability will add to the fundamental knowledge of human pluripotent stem cell (hPSC) culture, and may suggest new approaches for achieving a robust, reproducible, and scalable process that result in consistent product quality and yields. Here, the current state of the understanding of the fundamental mechanisms that govern the growth kinetics of hPSCs between static and dynamic cultures is reviewed, the factors causing fluctuations are identified, and culture strategies that might eliminate or minimize the occurrence of cell‐to‐cell variability arising from these fluctuations are discussed. The existing challenges in the development of hPSC expansion methods for enabling the transition from process development to large‐scale production are addressed, a mandatory step for industrial and clinical applications of hPSCs. 相似文献