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101.
Y Katagata K Aso M Sato T Yoshida 《Biochemical and biophysical research communications》1992,182(3):1440-1445
To date, the largest keratin peptide(K1, 68 KD) has been absent in cultured human squamous cell carcinomas. Using a low salt aqueous solution, not containing high salt and Triton X-100, as a washing buffer for keratin extraction, followed by two dimensional polyacrylamide gel electrophoresis, immunological techniques and Northern blot analysis, we demonstrated K1 peptide in two kinds of cultured human squamous cell carcinomas. Until now keratin extraction has been done using high salt/Triton X-100 solution during which K1 peptide may be removed together developed an affinity with the buffer. Many investigators may have therefore overlooked K1. 相似文献
102.
H Nakayama Y Hatanaka E Yoshida K Oka M Takanohashi Y Amano Y Kanaoka 《Biochemical and biophysical research communications》1992,184(2):900-907
Forty three percent of the labeled sites, at least, in the electroplax sodium channel with a photoactivable tetrodotoxin derivative were identified by probing protease-digested labeled fragments with several sequence-directed antibodies. They are located in the loop between segments S5 and S6 of domain IV, as well as the region containing transmembrane segment S6 and adjacent extracellular and cytoplasmic sequences in domain III. No photolabeled fragments were detected in the corresponding region of domain I. These results suggest that C-11 of tetrodotoxin where the photoreactive moiety is attached orients to the region between S5 and S6 in domain III and IV. Probable orientation of the tetrodotoxin molecule in sodium channels is considered by taking together with the recent report of the site-directed mutagenesis. 相似文献
103.
Gene expression of metalloproteinase and its inhibitor in mesangial cells exposed to high glucose. 总被引:3,自引:0,他引:3
M Kitamura A Kitamura T Mitarai N Maruyama R Nagasawa T Kawamura H Yoshida T Takahashi O Sakai 《Biochemical and biophysical research communications》1992,185(3):1048-1054
To clarify the roles of metalloproteinases and their inhibitor (TIMP) in diabetic glomerulopathy, we studied the effect of a high glucose concentration on the gene expression of metalloproteinase transin and TIMP as well as collagen type IV and laminin in cultured rat mesangial cells (MCs). In the high glucose group, collagen type IV, laminin, and TIMP mRNA levels were all elevated in a concentration-dependent manner, whereas transin expression was suppressed. Osmotic control of high glucose with mannitol selectively stimulated TIMP expression. We hypothesize that high glucose decreases matrix-degrading activity as well as increases matrix productivity in MCs. 相似文献
104.
Tadao Hashimoto Yukuo Yoshida Kunio Tagawa 《Journal of bioenergetics and biomembranes》1990,22(1):27-38
An intrinsic ATPase inhibitor inhibits the ATP-hydrolyzing activity of mitochondrial F1F0-ATPase and is released from its binding site on the enzyme upon energization of mitochondrial membranes to allow phosphorylation of ADP. The mitochondrial activity to synthesize ATP is not influenced by the absence of the inhibitor protein. The enzyme activity to hydrolyze ATP is induced by dissipation of the membrane potential in the absence of the inhibitor. Thus, the inhibitor is not responsible for oxidative phosphorylation, but acts only to inhibit ATP hydrolysis by F1F0-ATPase upon deenergization of mitochondrial membranes. The inhibitor protein forms a regulatory complex with two stabilizing factors, 9K and 15K proteins, which facilitate the binding of the inhibitor to F1F0-ATPase and stabilize the resultant inactivated enzyme. The 9K protein, having a sequence very similar to the inhibitor, binds directly to F1 in a manner similar to the inhibitor. The 15K protein binds to the F0 part and holds the inhibitor and the 9K protein on F1F0-ATPase even when one of them is detached from the F1 part. 相似文献
105.
Ferredoxin-sulfite reductases (Fd-SiRs) [hydrogen-sulfide: ferredoxin oxidoreductase, EC 1.8.7.1] from leek leaves have been
purified to homogeneity. The enzymes (SiR 1, SiR 2 and SiR 3) were separated by Mono Q chromatography. The collective molecular
mass of the enzymes was estimated to be 65 kDa by gel filtration. In all three cases, subunit analysis by SDS-PAGE yielded
a single protein band corresponding to a molecular mass of 64 kDa, indicating that the enzymes each exist as a monomer. In
the oxidized forms, SiR 1, SiR 2 and SiR 3 all exhibited nearly identical absorption maxima at 279∼280, 389∼390, 588 and 714
nm, indicating that siroheme is involved in the catalysis of sulfite reduction. On enzymatic properties, SiR 1, SiR 2 and
SiR 3 could only react with the physiological electron donor, feriedoxin. The enzymes exhibited different heat stabilities.
The pH active curve obtained from SiR 2 was different from the others. Moreover, SiR 1 exhibited a lower Km value for ferredoxin
than SiR 2. Although the N-terminal sequence was the same, the results of some enzymatic properties, amino acid analysis,
and peptide mapping suggested the presence of the Fd-SiR isozymes in leek leaves. 相似文献
106.
Homogeneously purified poly(ADP-ribose) polymerase (PARP) specifically stimulated the activity of immunoaffinity-purified calf or human DNA polymerase by about 6 to 60-fold. Apparently, poly(ADP-ribosyl)ation of DNA polymerase was not necessary for the stimulation. The effects of PARP on DNA polymerase were biphasic: at very low concentrations of DNA, it rather inhibited its activity, whereas, at higher DNA concentrations, PARP greatly stimulated it. The autopoly(ADP-ribosyl)ation of PARP suppressed both its stimulatory and inhibitory effects. By immunoprecipitation with an anti-DNA polymerase antibody, it was clearly shown that PARP may be physically associated with DNA polymerase . Stimulation of DNA polymerase may be attributed to the physical association between the two, rather than to the DNA-binding capacity of PARP, since the PARP fragment containing only the DNA binding domain showed little stimulatory activity. The existence of PARP-DNA polymerase complexes were also detected in crude extracts of calf thymus. 相似文献
107.
Takayoshi Yoshida Hiroshi Watari 《European journal of applied physiology and occupational physiology》1994,69(6):465-473
To investigate the splitting of the inorganic phosphate (Pi) peak during exercise and recovery, a time-resolved 31phosphorus nuclear magnetic resonance spectroscopy (31P-MRS) technique was used. Seven healthy young sedentary male subjects performed knee flexion exercise in the prone position inside a 2.1-T magnet, with the surface coil for 31P-MRS being placed on the biceps femoris muscle. After a 1-min warm-up without loading, the exercise intensity was increased by 0.41 W at 15-s intervals until exhaustion, followed by a 5-min recovery period. The 31P-MRS were recorded every 5 s during the rest-exercise-recovery sequence. Computer-aided contour analysis and pixel imaging of the Pi and phosphocreatine peaks were performed. Five of the seven subjects showed two distinct Pi peaks during exercise, suggesting two different pH distributions in exercising muscle (high pH and low pH region). In these five subjects, the high-pH increased rapidly just after the onset of exercise, while the low-pH peak increased gradually approximately 60 s after the onset of exercise. During recovery, the disappearance of the high-pH peak was more rapid than that of the low-pH peak. These findings suggest that our method 31P-MRS provides a simple approach for studying the kinetics of the Pi peak and intramuscular pH during exercise and recovery. 相似文献
108.
Soh Chang-Ho; Kamiya Yuji; Yoshida Shigeo; Yamane Hisakazu; Takahashi Nobutaka 《Plant & cell physiology》1994,35(7):1037-1042
Oryzains, cysteine proteinases of rice seeds, are induced byGA3 in germinating rice seeds [Abe et al. (1987) Agric. Biol.Chem. 51: 1509]. The effects of GA1, GA3, GA4, GA9, and GA20on the production of oryzain and -amylase were investigatedin embryoless half- and whole-seeds of rice (cv. Nipponbare).When gibberellins (GAs) were incubated with embryoless half-seeds,GA1, GA3 and GA4 induced oryzain and -amylase, but GA9, andGA20 did not. GA9 and GAM induced oryzain and -amylase productionin whole seeds, but this production was inhibited by the simultaneousapplication of prohexadione, an inhibitor of 2ß- and3ß-hydroxylation of GAs. Prohexadione did not inhibitthe activities of oryzain and -amylase induced by GA1. Theseresults suggest that GAs possessing the 3ß-hydroxylgroup induce activities of oryzain and -amylase in rice seedsand that GA9 and GA20 have activity only after they are convertedmetabolically to active GAs, probably GA4 and GA1, respectively.GA1, was more active than GA4 in both half seeds and wholeseeds incubation. Oryzain and -amylase activities induced byGA4 were significantly inhibited in the presence of 104M prohexadione. This suggests that the conversion of GA1, toGA4 (13-hydroxylation) might be inhibited at a high dose ofprohexadione in whole seeds.
4Present address: Institute of Food Development, Kyung Hee University,Suwon 449-701, Korea 相似文献
109.
Summary Silica glass-entrapped lipase was prepared by the sol-gel method using tetramethoxysilane, and its esterification activity in n-hexane was examined for isoamylbutyrate formation. The hydrogel preparation containing a large amount of water exhibited enough activity. Although the activity of xerogel-entrapped lipase drastically decreased probably due to shrinkage of the gel matrix, the lyophilized gel retained much higher activity than the air-dried gel. 相似文献
110.
A cis-acting element and a trans-acting factor involved in the wound-induced expression of a horseradish peroxidase gene 总被引:2,自引:0,他引:2
Akiyoshi Kawaoka Tomohiro Kawamoto Masami Sekine Kazuya Yoshida Mitsuo Takano Atsuhiko Shinmyo 《The Plant journal : for cell and molecular biology》1994,6(1):87-97
The mechanisms that control the wound-induced expression of the prxC2 gene for horseradish peroxidase (HRP) have been investigated. Analysis of the regulatory properties of 5′-deleted promoters showed that a positive element involved in the response to wounding was located between −307 and −99 bp from the site of initiation of translation. In in vitro binding assays of tobacco nuclear proteins and DNA fragments of prxC2 promoter, the binding site was the Box 1 from −296 to −283 containing the CACGTG motif. To identify the functional role of Box 1, the prxC2 promoter that has been digested from the 5′ end to −289 with a disrupted Box 1 was fused to a reporter gene for β-glucuronidase (GUS). No induction of GUS activity was observed in transgenic tobacco plants with the prxC2(−289)/GUS construct. These data indicated that the expression of prxC2 in response to wounding required the Box 1 sequence from −296 to −283. Furthermore, a tobacco cDNA expression library was screened and a cDNA clone for a protein, designated TFHP-1, that bound specifically to the Box 1 sequence was identified. The putative TFHP-1 protein contains a basic region and leucine zipper (bZip) motif and a helix—loop—helix (HLH) motif. The mRNA for TFHP-1 was abundant in roots and stems, and it was not induced by wounding in leaves. In tobacco protoplasts, antisense TFHP-1 suppressed the expression of prxC2 (−529)/GUS. 相似文献