全文获取类型
收费全文 | 19737篇 |
免费 | 1541篇 |
国内免费 | 910篇 |
出版年
2024年 | 34篇 |
2023年 | 170篇 |
2022年 | 321篇 |
2021年 | 665篇 |
2020年 | 463篇 |
2019年 | 563篇 |
2018年 | 713篇 |
2017年 | 570篇 |
2016年 | 825篇 |
2015年 | 1192篇 |
2014年 | 1376篇 |
2013年 | 1542篇 |
2012年 | 1804篇 |
2011年 | 1675篇 |
2010年 | 1072篇 |
2009年 | 833篇 |
2008年 | 1122篇 |
2007年 | 935篇 |
2006年 | 885篇 |
2005年 | 775篇 |
2004年 | 716篇 |
2003年 | 710篇 |
2002年 | 570篇 |
2001年 | 312篇 |
2000年 | 282篇 |
1999年 | 277篇 |
1998年 | 157篇 |
1997年 | 117篇 |
1996年 | 94篇 |
1995年 | 104篇 |
1994年 | 100篇 |
1993年 | 74篇 |
1992年 | 109篇 |
1991年 | 94篇 |
1990年 | 68篇 |
1989年 | 66篇 |
1988年 | 63篇 |
1987年 | 45篇 |
1986年 | 47篇 |
1985年 | 48篇 |
1984年 | 48篇 |
1983年 | 36篇 |
1982年 | 27篇 |
1979年 | 30篇 |
1976年 | 24篇 |
1975年 | 28篇 |
1974年 | 33篇 |
1973年 | 38篇 |
1972年 | 33篇 |
1969年 | 24篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
101.
Comparison of DMO and flow cytometric methods for measuring intracellular pH and the effect of hyperthermia on the transmembrane pH gradient 总被引:1,自引:0,他引:1
Intracellular pH (pHi) was measured in both unheated and heated cells by the distribution of the weak acid, 5,5-dimethyl-2,4-oxazolidinedione-2-14C (14C-DMO), and by the fluorescence intensity ratio (I530/I630) of the pH sensitive fluorescent dye, 2',7'-bis(carboxyethyl)-5,6-carboxy-fluorescein (BCECF), analyzed by flow cytometry (FCM). BCECF-loaded Chinese hamster ovary (CHO) cells were analyzed by FCM after they had incubated in fresh medium at 37 degrees C for 90 min, during which time a decrease in fluorescence ratio stabilized. After stabilization, the pHi determined for CHO cells by the FCM method at pHe values of 6.0-8.1 agreed-within 0.1 pH units with that determined by the 14C-DMO method. There is a pH gradient across the plasma membrane that is not affected by heat. In CHO cells, the gradient, determined by DMO and FCM, is less or greater than pHe by 0.30 and 0.15 pH units at pHe 7.4 and 6.3, respectively, and in NG108-15 cells, the gradient determined by DMO increases to 0.50 pH units at pHe 6.3. Both cells maintained their pH gradients for at least 4 h after heating, although 99.9% of the cells were reproductively dead (survival of 10(-3)) after heating at 45.5 degrees C either at the normal pHe of 7.4 or at a low pHe of 6.4-6.7. 相似文献
102.
103.
A. A. J. J. L. Rutten B. G. A. G. G. Béquet-Passelecq H. B. W. M. Koëter 《In vitro cellular & developmental biology. Plant》1990,26(4):353-360
Summary A new method was developed for rabbit skin organ culture. In a two-compartment model, skin discs were cultured on a Millicell-HA
insert unit with a microporous membrane which allows transport of culture medium via the dermis into the epidermis, whereas
the epidermal side remains free of direct contact with culture medium. In this relatively simple two-compartment organ culture
model, rabbit skin could be cultured for 7 d in RPMI 1640 medium supplemented with fetal bovine serum, or for 2 d in RPMI
1640 medium supplemented with cofactors. The histomorphology and ultrastructure of 7-d cultured rabbit skin discs was essentially
similar to that of freshly isolated rabbit skin. Keratinocytes in the stratum basale continued to divide during organ culture.
The terminal differentiation of the epidermis continued in vitro as was found by the presence of keratohyalin granules, the
intact stratum corneum, and keratin expression. Furthermore, glucose consumption continued until culture Day 7, but thereafter
it declined rapidly. Concomitantly, degenerative changes were found. At the end of the 7-d culture period the distance between
single dermal collagen fibrils had increased as compared to noncultured skin. This model of skin organ cultures can be used
to study biological processes, dermal toxicity, and penetration and metabolism of xenobiotics in intact skin. Furthermore,
within certain limits, processes responsible for repair and regeneration of damaged skin can also be studied in this model
because the rabbit skin can be cultured for 7 d.
The present study was financially supported by grants of Duphar B. V. (Weesp, Netherlands), the European Community, and the
Dutch animal welfare organizations Samenwerkingsverband van de Nederlandse Vereniging tot Bescherming van Dieren en de Nederlandse
Bond tot Bestrijding van de Vivisectie, Anti-Vivisectie Stichting en Stichting Schoonheid Zonder Wreedheid. 相似文献
104.
Akira Yamada Masaaki Murakami Kenichi Ijima Hideo Yagita Ko Okumura Sakuzo Komatsu Toshimitsu Uede 《Microbiology and immunology》1996,40(7):513-518
The immunosuppressant FK506 prolongs allograft survival. However, at therapeutic doses it has significant side effects. A fusion protein consisting of the extracellular portion of CTLA4 and the Fc portion of human IgG (CTLA4IgG) also prolongs allograft survival, but large doses of CTLA4IgG are required for the induction of cardiac allograft acceptance. Therefore, we constructed a pentameric form of a new CTLA4 fusion protein, CTLA4IgM. We tested whether low doses of CTLA4IgG or CTLA4IgM in combination with subtherapeutic doses of FK506 can prolong allograft survival in a synergistic fashion. C57BL/6 (H-2b) neonatal hearts were transplanted to CBA/J (H-2k) mice in a heterotopic, nonvascularized cardiac allograft model. The findings demonstrate that a combination of low doses of FK506 plus a pentameric form of CTLA4Ig, CTLA4IgM, leads to significant graft survival, while a combination of FK506 plus CTLA4IgG does not. 相似文献
105.
Yeon-Jeong Kim Jong-Hoon Park Koung-Sook Kim Ji-Eun Chang Jeong Heon Ko Myung-Hee Kim Dong-Hyo Chung Tae-Wha Chung In-Seong Choe Young-Choon Lee Cheorl-Ho Kim 《Gene》1996,170(2):281-283
We have isolated and characterized the immediate (1651 bp) 5′-flanking region of the gene (GnT-III) encoding N-acetylglucosaminyltransferase III (GnT-III) from a human placental genomic library. Analysis of promoter elements shows a similarity to the 5′-flanking region of murine 1,4-galactosyltransferase. The sequence lacks obvious TATA elements and CCAAT boxes; however, putative regulatory sites, including 2 potential cAMP-response regulatory elements (CRE), 11 insulin-response element consensus sequences (IRE), 7 potential AP-2-binding sites, 2 SP1 consensus sequences (GC boxes) and 2 sequences similar to the half-palindromic glucocorticoid-responsive element (GRE), are present. 相似文献
106.
棕色固氮菌缺失nifZ基因的突变种固氮酶MoFe蛋白的纯化和性质 总被引:1,自引:0,他引:1
采用 52℃下加热 6 min,后经 DEAE- 52、Sephacryls S- 2 0 0和 Q- Sepharose等柱层析方法 ,分离纯化了棕色固氮菌 (Azotobacter vinelandii)缺失 nif Z基因突变种固氮酶 Mo Fe(Δnif Z Mo Fe)蛋白 ,其纯度达到电泳纯。Δnif Z Mo Fe蛋白的固氮活性为 2 83nmol C2 H2 还原 / (min·mg蛋白 ) ,远低于野生种 Mo Fe蛋白。Δnif Z Mo Fe蛋白对氧更敏感 ;热稳定性略低于野生种。Δnif Z Mo Fe蛋白的可见光吸收光谱与野生种 Mo Fe蛋白极为相似。其圆二色谱和磁圆二色谱在 450~ 550 nm与野生种 Mo Fe蛋白显著不同 ,表明其 P- cluster及其周围环境与野生种 Mo Fe蛋白有所差异。这亦可能是造成缺失 nif Z突变种 Mo Fe蛋白固氮活性低的原因。 相似文献
107.
Interactions between the ankyrin repeat-containing protein Akr1p and the pheromone response pathway in Saccharomyces cerevisiae. 总被引:4,自引:2,他引:2
下载免费PDF全文
![点击此处可从《Molecular and cellular biology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Akr1p, which contains six ankyrin repeats, was identified during a screen for mutations that displayed synthetic lethality with a mutant allele of the bud emergence gene BEM1. Cells from which AKR1 had been deleted were alive but misshapen at 30 degrees C and inviable at 37 degrees C. During a screen for mutants that required one or more copies of wild-type AKR1 for survival at 30 degrees C, we isolated mutations in GPA1, which encodes the G alpha subunit of the pheromone receptor-coupled G protein. (The active subunit of this G protein is G beta gamma, and G alpha plays an inhibitory role in G beta gamma-mediated signal transduction.) AKR1 could serve as a multicopy suppressor of the lethality caused by either loss of GPA1 or overexpression of STE4, which encodes the G beta subunit of this G protein, suggesting that pheromone signaling is inhibited by overexpression of Akr1p. Mutations in AKR1 displayed synthetic lethality with a weak allele of GPA1 and led to increased expression of the pheromone-inducible gene FUS1, suggesting that Akr1p normally (and not just when overexpressed) inhibits signaling. In contrast, deletion of BEM1 resulted in decreased expression of FUS1, suggesting that Bem1p normally facilitates pheromone signaling. During a screen for proteins that displayed two-hybrid interactions with Akr1p, we identified Ste4p, raising the possibility that an interaction between Akr1p and Ste4p contributes to proper regulation of the pheromone response pathway. 相似文献
108.
To clarify the way in which the light available for growth affectsrespiration in leaves of sun and shade plants, we examined therespiratory properties of mature leaves of Spinacia oleraceaL., a sun species, and of Alocasia macrorrhiza (L.) G. Don.,a shade species, that had been grown at various irradiances.In leaves of S. oleracea, the respiratory rates, on a dry massbasis, decreased with time during the night, and the higherwas the growth irradiance during the day, the higher was therespiratory rate. The marked decreases in the respiratory rateduring the night were accompanied by decreases in the concentrationof carbohydrates in the leaves. By contrast, the respiratoryrates of leaves of A. macrorrhiza were virtually constant throughoutthe night and the absolute rates were lower than those of S.oleracea even though the absolute value of the concentrationof carbohydrates and its decrease at night resembled to thosein S. oleracea. The maximum activities of respiratory enzymeswere also similar to those in S. oleracea. However, the leavesof A. macrorrhiza contained less soluble protein than thoseof S. oleracea. These results suggest that, in S. oleracea,the concentration of carbohydrates might determine the respiratoryrate while such is not the case in A. macrorrhiza. The lowerrespiratory rates in A. macrorrhiza might be due to a lowerdemand for ATP. (Received November 29, 1995; Accepted February 15, 1996) 相似文献
109.
Two Chains of Rhamnogalacturonan II Are Cross-Linked by Borate-Diol Ester Bonds in Higher Plant Cell Walls 总被引:21,自引:4,他引:17
下载免费PDF全文
![点击此处可从《Plant physiology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Polysaccharide moiety of the boron-polysaccharide complex (T. Matoh, K. Ishigaki, K. Ohno, J. Azuma [1993] Plant Cell Physiol 34: 639-642) isolated from radish (Raphanus sativus) roots has been shown to be rhamnogalacturonan II by glycosyl-linkage analysis and the presence of diagnostic monosaccharides, including apiose, aceric acid, 2-O-methylfucose, and 3-deoxy-D-manno-2-octulosonic acid. Removal of boron from the complex reduced the molecular weight by one-half without causing a significant increase in the number of reducing end groups, indicating that boron, as boric acid, links two rhamnogalacturonan II chains together to form the boron-polysaccharide complex. 相似文献
110.