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101.
The effect of frequency of transvaginal follicular aspiration on oocyte yield and subsequent superovulatory response was studied in 2 experiments. In Experiment 1, 32 primiparous Hereford x Friesian cows were assigned to 4 treatments (n = 8 per treatment). Oocyte recovery was carried out once a week for 12, 8, 4 or 0 (control) wk. Embryo recovery for all animals was 7 wk after the completion of the aspiration schedules. In Experiment 2, the effects of oocyte recovery once or twice a week (n = 8 per treatment; control n = 18) for 12 wk and response to superovulation 4 wk after the last aspiration were compared using nulliparous purebred Simmental heifers. Increasing the period of once weekly aspirations from 4 to 12 wk (Experiment 1) did not affect the number of follicles observed per session (mean +/- SEM; 10.0 +/- 0.82) or aspirated (7.8 +/- 0.71), but the recovery rate of oocytes from follicles aspirated was greater for donors aspirated for either 4 or 8 wk than for 12 wk (32.3 +/- 3.73 vs 28.4 +/- 2.61 vs 20.1 +/- 2.13 %; P < 0.05). Following the last aspiration and prior to commencing superovulatory procedures, estrus or estrous activity was observed in 7 8 , 8 8 , 7 8 and 6 8 of the animals aspirated over 12, 8, 4 or 0 wk, respectively. Subsequent superovulatory responses and in vivo embryo recoveries were similar for all aspiration treatments and for control animals. Changing the frequency of oocyte recovery from once to twice weekly (Experiment 2) did not affect the numbers of follicles observed (9.1 +/- 0.63 vs 8.3 +/- 0.85), follicles aspirated (5.9 +/- 0.56 vs 6.2 +/- 0.69), oocytes recovered (1.7 +/- 0.27 vs 1.9 +/- 2.0) per session or the oocyte recovery rate (29.4 +/- 2.4 vs 30.4 +/- 2.4 %); nor was there any effect of frequency of aspiration on subsequent superovulatory response and embryo recovery. In conclusion, increasing the period of aspiration from 4 to 12 wk and the frequency from once to twice a week over 12 wk did not reduce the number of follicles observed or aspirated, or number of oocytes recovered per donor per session. Subsequent estrous cyclicity and responses to superovulation were unaffected by the periods or frequencies of oocyte recovery examined here. 相似文献
102.
The estimation of the amount of evolutionary divergence that has taken
place between two DNA coding sequences depends strongly on the degree of
constraint on amino acid replacements. If amino acid replacements are
relatively unconstrained, the individual nucleotide is the appropriate unit
of analysis and the method of Tajima and Nei can be used. If amino acid
replacements are constrained, however, this method is shown to be
inapplicable. For sequences with strong amino acid constraints, a method is
outlined analogous to the Tajima and Nei method using codons as the unit of
analysis. Only synonymous substitutions are used. Codon usage data can be
employed to estimate the necessary parameters of the calculation, or a
priori models of substitution may be employed. Sequences with significant
but intermediate constraints on amino acid replacements are, in principle,
unanalyzable.
相似文献
103.
104.
A kinetic study has been carried out over the pH range of 2.63-9.37 for the reaction of horseradish peroxidase with hydrogen peroxide to form compound I of th;e enzyme. Analysis of the results, indicates that there are two kinetic influencing, ionizable groups on the enzyme with pKa values of 3.2 and 3.9. Protonation of these groups results in a decrease in the rate of reaction of the enzyme with H2O2. A previous study of the kinetics of cyanide binding to horseradish peroxidase (Ellis, W.D. & Dunford, H.B.: Biochemistry 7, 2054-2062 (1968)) has been extended to down to pH 2.55, and analysis of these results also indicates the presence of two kinetically important ionizable groups on the enzyme with pKa values of 2.9 and 3.9. 相似文献
105.
K M Dolman B A van de Wiel C M Kam J J Abbink C E Hack A Sonnenberg J C Powers A E von dem Borne R Goldschmeding 《FEBS letters》1992,314(2):117-121
Physiological inhibitors were tested for their in vitro interaction with neutrophil proteinase 3 (PR3). The major plasma proteinase inhibitor of PR3 is alpha 1AT. We have developed a radioimmunoassay (RIA) for quantitative detection of PR3-alpha 1AT complexes formed in vivo in inflammatory exudates such as synovial fluid and plasma from patients with sepsis. Levels of PR3-alpha 1AT complexes correlated significantly with levels of human neutrophil elastase (HNE)-alpha 1AT complexes. Thus, in vivo alpha 1AT not only protects against excessive HNE activity, but also against excessive PR3 activity. 相似文献
106.
C M Kam J E Kerrigan K M Dolman R Goldschmeding A E Von dem Borne J C Powers 《FEBS letters》1992,297(1-2):119-123
Various amino acid and peptide thioesters were tested as substrates for human proteinase 3 and the best substrate is Boc-Ala-Ala-Nva-SBzl with a kcat/Km value of 1.0 x 10(6) M-1.s-1. Boc-Ala-Ala-AA-SBzl (AA = Val, Ala, or Met) are also good substrates with kcat/Km values of (1-4) x 10(5) M-1.s-1. Substituted isocoumarins are potent inhibitors of proteinase 3 and the best inhibitors are 7-amino-4-chloro-3-(2-bromoethoxy)isocoumarin and 3,4-dichloroisocoumarin (DCI) with kobs/[I] values of 4700 and 2600 M-1.s-1, respectively. Substituted isocoumarins, peptide phosphonates and chloromethyl ketones inhibited proteinase 3 less potently than human neutrophil elastase (HNE) by 1-2 orders of magnitude. 相似文献
107.
Phosphatidylinositol-linked FcRIII mediates exocytosis of neutrophil granule proteins, but does not mediate initiation of the respiratory burst 总被引:7,自引:0,他引:7
T W Huizinga K M Dolman N J van der Linden M Kleijer J H Nuijens A E von dem Borne D Roos 《Journal of immunology (Baltimore, Md. : 1950)》1990,144(4):1432-1437
In this report, we present data on the activation of different neutrophil effector functions by two distinct Fc-gamma receptors, FcRII and FcRIII. We and others have shown previously that IgG-dependent activation of phagocytosis and superoxide generation is mediated via FcRII. IgG-dependent exocytosis of granule proteins was assessed with Staphylococcus aureus Oxford opsonized with human IgG or with IgG-coated latex. Both anti-FcRII mAb and anti-FcRIII-F(ab')2 mAb inhibited this release, whereas the combination of these mAb inhibited this process more strongly than either mAb alone. This indicates that both FcRII and FcRIII are involved in IgG-dependent release of granule proteins. Cross-linking of the receptors by anti-FcR mAb and F(ab')2 fragments of goat-anti-mouse-Ig showed again that both FcRII and FcRIII mediate lysozyme release, whereas cross-linking of a control antigen (CD67) did not. By measuring the release of elastase and lactoferrin, we found that cross-linking of either FcRII or FcRIII induced release of both azurophilic and specific granules. Under these conditions, we did not measure any activation of the respiratory burst. When FcRIII was removed by treatment of neutrophils with glycosylphosphatidylinositol-specific phospholipase C, the lysozyme release induced by cross-linking of FcRIII was lower than the release from control neutrophils, whereas the release induced by cross-linking of FcRII was similar. Therefore, we conclude that IgG-dependent activation of neutrophils follows two distinct pathways: one via transmembrane FcRII, activating both the NADPH oxidase and the release of granule proteins (as was demonstrated previously by us and by others), and the other via phosphatidylinositol-linked FcRIII, activating exocytosis of granule proteins. 相似文献
108.
Mitogenic activity of pituitary hormones on cell cultures of normal and carcinogen-induced tumor epithelium from rat mammary glands 总被引:5,自引:2,他引:3 下载免费PDF全文
Cell suspension containing normal or tumor epithelium were readily obtained by enzymatically digesting rat mammary glands from perphenazine-treated (prolactin-hypersecreting) cycling, female virgin animals or hormone- responsive mammary tumors from animal treated with dimethylbenzanthracene. Cell suspensions were fractioned into predominantly epithelial and predominantly stromal cells by their differential rates of attachment to culture dishes. Both normal mammary and tumor epithelial cells were characterized by the presence of specific cell-junctional complexes, desmosome-like structures, surface microvilli, and their ability to synthesize casein. Serum-dependent protease activity was greater in cultures derived from tumors, and cells from such cultures grew in agarose whereas those from the non-neoplastic gland did not. The addition of prolactin to the culture medium stimulated DNA synthesis in primary or secondary epithelial cultures from tumors, whereas additional insulin and hydrocortisone with prolactin were required for similar levels of DNA synthesis in cultures from non-neoplastic glands. The fraction of cells synthesizing DNA was, however, smaller than that with 10 percent serum measured in the same time period. Both growth hormone and epidermal growth factor stimulated DNA synthesis but to a lesser extent than did prolactin. Prolactin with hydrocortisone and insulin were relatively inactive in promoting DNA synthesis of the nonepithelial cells whereas pituitary fibroblast growth factor was more active. These mitogenic effects were obtained when the hormones were added to the medium at near physiological concentrations, and paralleled the known activities of the hormones in control of mammary gland growth and development in the rat. 相似文献
109.
A molecular and evolutionary study of the beta-globin gene family of the Australian marsupial Sminthopsis crassicaudata 总被引:1,自引:0,他引:1
Cooper SJ; Murphy R; Dolman G; Hussey D; Hope RM 《Molecular biology and evolution》1996,13(7):1012-1022
Beta-globin gene families in eutherians (placental mammals) consist of a
set of four or more developmentally regulated genes which are closely
linked and, in general, arranged in the order 5'-embryonic/fetal genes-
adult genes-3'. This cluster of genes is proposed to have arisen by tandem
duplication of ancestral beta-globin genes, with the first duplication
occurring 200 to 155 MYBP just prior to a period in mammalian evolution
when eutherians and marsupials diverged from a common ancestor. In this
paper we trace the evolutionary history of the beta-globin gene family back
to the origins of these mammals by molecular characterization of the
beta-globin gene family of the Australian marsupial Sminthopsis
crassicaudata. Using Southern and restriction analysis of total genomic DNA
and bacteriophage clones of beta-like globin genes, we provide evidence
that just two functional beta-like globin genes exist in this marsupial,
including one embryonic- expressed gene (S.c-epsilon) and one
adult-expressed gene (S.c-beta), linked in the order 5'-epsilon-beta-3'.
The entire DNA sequence of the adult beta-globin gene is reported and shown
to be orthologous to the adult beta-globin genes of the North American
marsupial Didelphis virginiana and eutherian mammals. These results,
together with results from a phylogenetic analysis of mammalian beta-like
globin genes, confirm the hypothesis that a two-gene cluster, containing an
embryonic- and an adult-expressed beta-like globin gene, existed in the
most recent common ancester of marsupials and eutherians. Northern analysis
of total RNA isolated from embryos and neonatals indicates that a switch
from embryonic to adult gene expression occurs at the time of birth,
coinciding with the transfer of the marsupial from a uterus to a pouch
environment.
相似文献
110.
Stebbings R Findlay L Edwards C Eastwood D Bird C North D Mistry Y Dilger P Liefooghe E Cludts I Fox B Tarrant G Robinson J Meager T Dolman C Thorpe SJ Bristow A Wadhwa M Thorpe R Poole S 《Journal of immunology (Baltimore, Md. : 1950)》2007,179(5):3325-3331
The CD28-specific mAb TGN1412 rapidly caused a life-threatening "cytokine storm" in all six healthy volunteers in the Phase I clinical trial of this superagonist, signaling a failure of preclinical safety testing. We report novel in vitro procedures in which TGN1412, immobilized in various ways, is presented to human white blood cells in a manner that stimulates the striking release of cytokines and profound lymphocyte proliferation that occurred in vivo in humans. The novel procedures would have predicted the toxicity of this superagonist and are now being applied to emerging immunotherapeutics and to other therapeutics that have the potential to act upon the immune system. Data from these novel procedures, along with data from in vitro and in vivo studies in nonhuman primates, suggest that the dose of TGN1412 given to human volunteers was close to the maximum immunostimulatory dose and that TGN1412 is not a superagonist in nonhuman primates. 相似文献