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951.
Bettina Schmidt Thomas Tradler Jens-U. Rahfeld Birgit Ludwig Bunty Jain Karlheinz Mann K. Peter Rücknagel Bernhard Janowski Angelika Schierhorn Gerhard Küllertz Jörg Hacker Gunter Fischer 《Molecular microbiology》1996,21(6):1147-1160
Legionella pneumophila is the causative agent of a severe form of pneumonia in humans (Legionnaires’disease). A major virulence factor, the Mip protein (FK506-binding protein, FKBP25mem), belongs to the enzyme family of peptidyl-prolyl cis/trans isomerases (PPIases). Here we show that L. pneumophila Philadelphia I possesses an additional cytoplasmic PPiase at a level of enzyme activity comparable to that of FKBP25mem. The N-terminal amino acid sequence of the purified protein was obtained by Edman degradation and showed that the protein is a member of the cyclophilin family of PPIases. The Icy gene (Legionella cycophn) was cloned and sequenced. It encodes a putative 164-amino-acid protein with a molecular mass of 17 968 Da called L. pneumophila cyclophilin 18 (L. p. Cyp18). Amino acid sequence comparison displays considerable similarity to the cytoplasmic and the periplasmic cyclophilins of Escherichia coll with 60.5% and 51.5% identity, respectively. The substrate specificity and inhibition by cyclosporin A revealed a pattern that is typically found for other bacterial cyclophilins. An L. pneumophila Cyp18 derivative with a 19-amino-acid polypeptide extension including a 6-histi-dine tag and an enterokinase cleavage site exhibits 相似文献
952.
Lene Jorgensen Anton P. J. Middelberg Brian K. O'Neill Connor J. Thomas 《Biotechnology Techniques》1996,10(2):83-88
Summary Slot- and dot-blotting are commonly used to evaluate levels of messenger ribonucleic acid (mRNA). Quantitation of bacterially-expressed chloramphenicol acetyl transferase (CAT) mRNA by this method is highly dependent on total RNA immobilised onto the solid support as well as mRNA concentration. mRNA quantitation by comparison with a pure standard results in underestimation. An improved protocol for CAT mRNA detection is described. 相似文献
953.
Torsten Müller Stefan Fiedler Thomas Schnelle Kai Ludwig Hartmut Jung Günter Fuhr 《Biotechnology Techniques》1996,10(4):221-226
Summary Combining dielectrophoretic and hydrodynamic forces in micro electrode structures allows enrichment and stable trapping of viruses in aqueous solutions. Fluorescently labelled Influenza and Sendai viruses were collected from solutions of 2*105 – 2*108 viruses/l within a few seconds. In the central part of the trap a virus aggregate of about 2–9 m in diameter was formed. This corresponds to a local enrichment of viruses up to a factor of about 1400. 相似文献
954.
955.
Dale L. Ludwig James R. Stringer David C. Wight Thomas C. Doetschman John J. Duffy 《Transgenic research》1996,5(6):385-395
The FLP recombinase of yeast catalyses site-specific recombination between repeated FLP recombinase target (FRT) elements in yeast and in heterologous system (Escherichia coli, Drosophila, mosquito and cultured mammalian cells). In this report, it is shown that transient FLP recombinase expression can recombine and activate an extrachromosomal silent reporter gene following coinjection into fertilized one-cell mouse eggs. Furthermore, it is demonstrated that introduction of a FLP-recombinase expression vector into transgenic one-cell fertilized mouse eggs induces a recombination event at a chromosomal FRT target locus. The resulting event occured at the one-cell stage and deleted a chromosomal tandem array of a FRT containinglacZ expression cassette down to one or two copies. These results demonstrate that the FLP recombinase can be utilized to manipulate the genome of transgenic animals and suggest that FLP recombinase-mediated plasmid-to-chromosome targeting is feasible in microinjected eggs. 相似文献
956.
Mark D. Sternlicht Suzi Safarians Thomas C. Calcaterra Sanford H. Barsky 《In vitro cellular & developmental biology. Animal》1996,32(9):550-563
Summary Myoepithelial cells exert important paracrine effects on epithelial morphogenesis and mitogenesis through direct cell-cell
interactions and through synthesis of a basement membrane extracellular matrix. To study these effects further, this study
established the first immortalized human myoepithelial cell line, HMS-1, and transplantable xenograft, HMS-X, from the rare
parotid basal cell adenocarcinoma. The cell line exhibited a fully differentiated myoepithelial phenotype and the xenograft
exhibited the rare property of accumulating an abundant extracellular matrix composed of both basement membrane and nonbasement
membrane components with the latter predominating. With HMS-1 as a feeder layer, dramatic and specific induction of epithelial
morphogenesis (sheroid formation) occurred with selected normal epithelial and primary carcinoma target cells. HMS-1 and HMS-X
provide distinct advantages over the conventional murine matrices in existence. They will be invaluable in future studies
of human tumor-myoepithelial and matrix interactions important for tumor cell growth, invasion, and metastasis. 相似文献
957.
Thomas J. Wiese Sherry A. Wuensch Paul D. Ray 《Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology》1996,114(4):417-422
Rabbit, pigeon and rat liver mitochondria convert exogenous phosphoenolpyruvate and acetylcarnitine to citrate at rates of 14, 74 and 8 nmol/15 min/mg protein. Citrate formation is dependent on exogenous HCO3, is increased consistently by exogenous nucleotides (GDP, IDP, GTP, ADP, ATP) and inhibited strongly by 3-mercaptopicolinate and 1,2,3-benzenetricar☐ylate. Citrate is not made from pyruvate alone or combined with acetylcarnitine. Pigeon and rat liver mitochondria make large amounts of citrate from exogenous succinate, suggesting the presence of an endogenous source of acetyl units or a means of converting oxalacetate to acetyl units. Citrate synthesis from succinate by pigeon and rabbit mitochondria is increased significantly by exogenous acetylcarnitine. Pigeon and rat liver contain 80 and 15 times, respectively, more ATP:citrate lyase activity than does rabbit liver. Data suggest that mitochondrial phosphoenolpyruvate car☐ykinasein vivo could convert glycolysis-derived phosphoenolpyruvate to oxalacetate that, with acetyl CoA, could form citrate for export to support cytosolic lipogenesis as an activator of acetyl CoA car☐ylase, a carbon source via ATP:citrate lyase and NADPH via NADP: malate dehydrogenase or NADP: isocitrate dehydrogenase. 相似文献
958.
Michael Linden Ph.D. Thomas Habib Vesna Radojevic 《Applied psychophysiology and biofeedback》1996,21(1):35-49
Eighteen children with ADD/ADHD, some of whom were also LD, ranging in ages from 5 through 15 were randomly assigned to one of two conditions. The experimental condition consisted of 40 45-minute sessions of training in enhancing beta activity and suppressing theta activity, spaced over 6 months. The control condition, waiting list group, received no EEG biofeedback. No other psychological treatment or medication was administered to any subjects. All subjects were measured at pretreatment and at posttreatment on an IQ test and parent behavior rating scales for inattention, hyperactivity, and aggressive/defiant (oppositional) behaviors. At posttreatment the experimental group demonstrated a significant increase (mean of 9 points) on the K-Bit IQ Composite as compared to the control group (p<.05). The experimental group also significantly reduced inattentive behaviors as rated by parents (p<.05). The significant improvements in intellectual functioning and attentive behaviors might be explained as a result of the attentional enhancement affected by EEG biofeedback training. Further research utilizing improved data collection and analysis, more stringent control groups, and larger sample sizes are needed to support and replicate these findings.This research was supported by an equipment grant by Autogenics Systems. Portions of this paper were presented at the annual convention of the Association of Applied Psychophysiology and Biofeedback, March, 1993 in Los Angeles and at the annual meeting of the Biofeedback Society of California, November, 1992 in Monterey, California. The authors gratefully acknowledge Todd Fischer and Paul Clopton for their valuable assistance in statistical analysis for this article. 相似文献
959.
Thomas Hiltonen Jan Karlsson Kristin Palmqvist Adrian K. Clarke Göran Samuelsson 《Planta》1995,195(3):345-351
An intracellular carbonic anhydrase (CA; EC 4.2.1.1) was purified and characterised from the unicellular green alga Coccomyxa sp. Initial studies showed that cultured Coccomyxa cells contain an intracellular CA activity around 100 times higher than that measured in high-CO2-grown cells of Chlamydomonas reinhardtii CW 92. Purification of a protein extract containing the CA activity was carried out using ammonium-sulphate precipitation followed by anion-exchange chromatography. Proteins were then separated by native (non-dissociating) polyacrylamide gel electrophoresis, with each individual protein band excised and assayed for CA activity. Measurements revealed CA activity associated with two discrete protein bands with similar molecular masses of 80 +5 kDa. Dissociation by denaturing polyacrylamide gel electrophoresis showed that both proteins contained a single polypeptide of 26 kDa, suggesting that each 80-kDa native protein was a homogeneous trimer. Isoelectric focusing of the 80-kDa proteins also produced a single protein band at a pH of 6.5. Inhibition studies on the purified CA extract showed that 50% inhibition of CA activity was obtained using 1 M azetazolamide. Polyclonal antibodies against the 26-kDa CA were produced and shown to have a high specific binding to a single polypeptide in soluble protein extracts from Coccomyxa cells. The same antiserum, however, failed to cross-react with soluble proteins isolated from two different species of green algae, Chlamydomonas reinhardtii and Chlorella vulgaris. Correspondingly, antisera directed against pea chloroplastic CA, extracellular CA from C. reinhardtii and human CAII, showed no cross-hybridisation to the 26-kDa polypeptide in Coccomyxa. The 26-kDa protein was confirmed as being a CA by N-terminal sequencing of two internal polypeptide fragments and alignment of these sequences with that of previously identified CA proteins from several different species.Abbreviations CA
carbonic anhydrase
- CCM
CO2-concentrating mechanism
- IEF
isoelectric focusing
- Rubisco
ribulose-l,5-bisphosphate carboxylase/oxygenase
We would like to thank Drs. Cecilia Forsman, Inga-Maj Johansson and Nalle Jonsson for their valuable advice concerning the isolation of CA. This work was supported by the Swedish Natural Research Council and Seth M. Kempes Memorial foundation. 相似文献
960.
Thomas Geissmann 《Primates; journal of primatology》1995,36(3):447-455
Data concerning the status, habitat, and vocalizations of yellow-cheeked crested gibbons (Hylobates gabriellae) were collected during a short field trip to the Nam Bai Cat Tien National Park (southern Vietnam). Nam Bai Cat Tien may
be the southernmost locality where crested gibbons (i.e. theHylobates concolor group) still survive. Fewer songs were heard at Nam Bai Cat Tien National Park than at other crested gibbon sites visited
by the author. At least two gibbon groups appear to have been greatly reduced in number since previous surveys in the park.
There is some evidence that both the gibbon population and the gibbon habitat in Nam Bai Cat Tien are disturbed. The first
case of a great call solo song in wild gibbons of theconcolor group is reported. Great calls ofH. gabriellae are described and documented with sonagrams for the first time. They differ from those previously described forH. leucogenys. 相似文献