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21.
J L Colbran S H Francis A B Leach M K Thomas H Jiang L M McAllister J D Corbin 《The Journal of biological chemistry》1992,267(14):9589-9594
Bovine lung cGMP-binding cGMP-specific phosphodiesterase (cG-BPDE) is a potent and relatively specific substrate for cGMP-dependent protein kinase (cGK) as compared to cAMP-dependent protein kinase (cAK) (Thomas, M. K., Francis, S. H., and Corbin, J. D. (1990) J. Biol. Chem. 265, 14971-14978). A synthetic peptide, RKISASEFDRPLR (BPDEtide), was synthesized corresponding to the sequence surrounding the phosphorylation site in cG-BPDE. BPDEtide retained the cGK/cAK kinase specificity demonstrated by native cG-BPDE: the apparent Km of BPDEtide for cGK was 5-fold lower than that for cAK (Km = 68 and 320 microM, respectively). Vmax values were 11 mumol/min/mg for cGK and 3.2 mumol/min/mg for cAK. The peptide was not phosphorylated to a measurable extent by protein kinase C or by calcium/calmodulin-dependent protein kinase II. Thus, the primary amino acid sequence of the peptide substrate was sufficient to confer kinase specificity. Studies in crude tissue extracts indicated that BPDEtide was the most selective peptide substrate documented for measuring cGK activity. Peptide analogs of BPDEtide were synthesized to determine the contribution of specific residues to cGK or cAK substrate specificity. Substitution of a Lys for the amino-terminal Arg did not reduce cGK/cAK specificity; neither did the exchange of an Ala for the non-phosphorylated Ser nor the removal of the 3 carboxyl-terminal residues. A truncated BPDEtide (RKISASE) served equally well as substrate (Km approximately 90 microM) for both kinases. However, restoration of the Phe, to yield RKISASEF, reproduced the original cGK/cAK specificity for BPDEtide (Km = 120 and 480 microM, respectively), primarily by decreasing the affinity of cAK. Addition of a carboxyl-terminal Phe to the peptide RKRSRAE (derived from the sequence of the cGK phosphorylation site in histone H2B) or to the peptide LRRASLG (derived from the sequence of the cAK phosphorylation site in pyruvate kinase) also improved the cGK/cAK specificity by decreasing the affinity of cAK. These data suggested that the Phe in each substrate tested is a negative determinant for cAK. 相似文献
22.
During a 13-month period, 64 lactating dairy cows of 2 genetic lines, Holstein and crossbred, housed indoors year-round were subjected to 2 superovulations and embryo collections within 112 days post partum. Half of the follicle stimulating hormone (FSH) treatments were given in a descending dosage regimen (Treatment A; 6.5 mg, 5.5 mg, 4.5 mg, and 3.5 mg, twice a day, total 40 mg) over 4 days; the remaining half of the treatments were administered in a constant dosage regimen (Treatment B) of 5 mg twice a day over 4 days. There were no significant differences due to treatment in the number of cows stimulated (more than 2 corpora lutea) nor in the number of ova/embryos collected. However, embryos were obtained from more cows (P<0.05) when treated with the descending dosage regimen. More cows (P<0.05) were stimulated by the superovulatory treatment during the first period than during the second period regardless of the regiment used, treatment A or B. More embryos (P<0.05) were obtained from the Holstein line than from the crossbred line. Fifty-two cows were inseminated at least once after the second embryo collection. Overall, 41 cows (79%) became pregnant after the second collection, requiring up to 4 services. These results suggest that the reproduction of dairy cattle housed indoors year-round is not adversely affected by 2 superovulation treatments and embryo collections within 112 days post partum. The question as to whether the administration of FSH is more efficacious in a descending dosage regimen or a constant dosage regimen was not resolved. 相似文献
23.
Coccidian oöcysts recovered from the faeces of rough earth snakes Virginia striatula (Serpentes: Colubridae) were found to represent two previously unreported eimerians. Oöcysts of Eimeria desotoensis n. sp. were found in 5/32 (16%) of the snakes and were spherical to ellipsoidal, 18.4 × 17.2 (15–21.5 × 15–19.5) μm, with a thin, single-layered wall; their shape-index (length/width) was 1.07 (1.00–1.23). A micropyle and oöcyst residuum were absent; polar granule were present in 33% of the oöcysts. The sporocysts were ovoidal, 11.5 × 7.6 (10.5–13 × 7–8) μm, with a Stieda body; their shape-index was 1.51 (1.30–1.68). The sporocyst residuum was moderate in size and composed of a cluster of granules. Oöcysts of Eimeria hobartsmithi n. sp. were found in 2/32 (6%) of the snakes and were subspherical to ellipsoidal, 18.0 × 15.7 (16–20 × 15–17) μm, with a thin, single-layered wall; their shape-index was 1.15 (1.02–1.32). A micropyle, oöcyst residuum and polar granule were absent. The sporocysts were elongate, 13.2 × 6.3 (12–14.5 × 6–6.5) μm, with a Stieda body; their shape-index was 2.10 (1.88–2.34). A large sporocyst residuum was present in each sporocyst, often obscuring the sporozoites. In addition to the two new species, oöcysts of E. striatula Upton & McAllister, 1990 were observed in 38% of the snakes. 相似文献
24.
Exercise training-induced coronary vascular adaptation. 总被引:5,自引:0,他引:5
Aerobic exercise training induces an increase in coronary vascular transport capacity. This increased transport capacity is the result of increases in both blood flow capacity and capillary exchange capacity. These functional changes are the result of two major types of adaptive responses, structural vascular adaptation and altered control of vascular resistance. Structural vascular adaptation occurs in response to exercise training in at least two forms, increases in the cross-sectional area of the proximal coronary arteries and angiogenesis. Angiogenesis has been demonstrated in that training causes moderate cardiac hypertrophy while maintaining or increasing capillary density and increasing arteriolar density. Training-induced changes in coronary vascular control have been shown to include altered coronary responses to vasoactive substances, changes in endothelium-mediated vasoregulation, and alterations in the cellular-molecular control of intracellular free Ca2+ in both endothelial and vascular smooth muscle cells isolated from coronary arteries of exercise-trained animals. The signal or signals for these adaptive responses remain unknown. The hypothesis that the adaptive strategy entails maintenance of normal shear stress in coronary arterial vessels is discussed. We propose that as a result of training-induced structural vascular adaptations and alterations in the control of vascular resistance, shear stress throughout the coronary vasculature is returned to the level present in sedentary animals. The signal for adaptation may be peak shear stress during exercise and/or average shear stress over a 24-h period of time. 相似文献
25.
The baboon endogenous virus genome. II. Provirus sequence variations in baboon cell DNA. 总被引:7,自引:2,他引:5 下载免费PDF全文
M Cohen N Davidson R V Gilden R M McAllister M O Nicolson R M Stephens 《Nucleic acids research》1980,8(19):4423-4440
Restriction analysis of the approximately 100 integrated baboon endogenous virus (BaEV) proviruses in baboon cells and tissues has revealed two major sequence variations, both in the gag gene region of the genome. One, a 150 nucleotide pair insert, is present in a small proportion of the proviral DNAs and some baboons, but is present in the majority of the proviral DNAs of other baboons. The second, a Bam HI recognition sequence located 2.25 kb from the proviral 5' end, is missing or modified in approximately one-half of the integrated genomes. We consider the possibility that accumulation of proviruses not containing the 0.15 kb insert is correlated with viral activation and expression since it is this form that is a replication intermediate in freshly infected permissive cells. It is evident from these initial studies that the organization of the multiple BaEV proviruses in baboon DNA has undergone modification during evolution. 相似文献
26.
A study has been made to determine if the cytotoxicity observed when cells in culture were exposed to liposome-entrapped cytotoxic drugs was liposome mediated or resulted from leakage of drug from the liposomes with subsequent uptake of free drug by the cells. In preliminary experiments with the EMT6 cell line in monolayer culture, the cytotoxicity observed when the cells were exposed to a range of concentrations of liposome-entrapped methotrexate, actinomycin D and cytosine arabinoside for a variety of liposome compositions was somewhat less than that observed when the cells were exposed to similar concentrations of free drug. We suspected that the cytotoxicity was mediated via uptake of free drug leaked from liposomes. This was confirmed in experiments involving the EMT6 and S49 cell lines in monolayer or suspension culture, respectively, in the absence and presence of the nucleoside transport inhibitor, 6-((4-nitrobenzyl)thio)-9-β-d-ribofuranosylpurine. Additional experiments were performed on a transport-deficient mutant of the S49 cell line, the AE1 cell line. No evidence for liposome-mediated cell death could be found in these cell lines when tubercidin 5′-monophosphate was entrapped in either large or small unilamellar liposomes composed of egg phosphatidylcholine/ cholesterol (2:1), bovine brain phosphatidylserine/egg phosphatidylcholine/ cholesterol (8:2:5) or egg phosphatidylcholine/stearylamine/cholesterol (10:1:5). Considerable toxicity due to empty liposomes of a variety of compositions was observed in the S49 cell line at high lipid concentrations. 相似文献
27.
Net genetic and temporary epistatic or maternal environmental responses to selection for egg production in chickens 下载免费PDF全文
The difference between progeny from selected and unselected parents (T) was experimentally partitioned into net genetic change (GA), temporary favorable epistatic combinations (GEP) and egg-transmitted maternal environment (M) in two strains of Leghorns selected over 14 years for early pure-strain egg production. Differences among progeny from selected sires and dams, selected sires only, selected dams only, unselected sires and dams and the parental generation were equated to expected GA, GEP and M responses for each trait. Total response was 3.3% for early egg number, 3.7% for total egg number, 0.5% for egg weight, 3.8% for early egg mass and 4.2% for total egg mass. Among progeny that survived the test period and were judged to be normal, total response was 2.6% for total number of eggs, 3.0% for early egg mass and 3.1% for total egg mass. The percentage of T attributed to GA was 9% for early egg number, 24% for total egg number, 43% for early egg mass and 47% for total egg mass; but 52% for total egg number, 98% for early egg mass and 71% for total egg mass of normal survivors. Temporary maternal selection responses (M) were (1) positive for number of eggs and egg masses, (2) greater for all progeny than for normal survivors, and (3) increased with progeny age. The results suggest that M was caused by reduced egg-transmitted disease. Epistatic selection response was positive for earlier sexual maturity and for number of eggs, but negative for egg weight and thus was small for egg masses. Temporary epistatic and maternal responses can explain overestimation of additive genetic response from offspring-parent regression or from replicated single-generation selection and apparent superiority of mass selection over family or combined selection. 相似文献
28.
Comparative Membrane Microviscosity of Fish and Mammalian Rhabdoviruses Studied by Fluorescence Depolarization 下载免费PDF全文
Norman F. Moore Yechezkel Barenholz Philip E. McAllister Robert R. Wagner 《Journal of virology》1976,19(1):275-278
The microviscosity of the hydrophobic region of the membrane of infectious hematopoietic necrosis virus was determined using fluorescence depolarization analysis of the probe 1,6-diphenyl-1,3,5-hexatriene and was found to be much lower at 37 C than that of another rhabdovirus, vesicular stomatitis virus. However, the microviscosity of this fish virus at 18 C, the temperature at which it was grown, corresponded to the microviscosity of vesicular stomatitis virus at 37 C. Data obtained with the fish virus host cell (chinook salmon embryo cells) grown at 18 C suggest that its membranes have a lower microviscosity than either L-929 or BHK-21 cells (the vesicular stomatitis virus host cells) grown at 37 C. 相似文献
29.
Comparative studies on the structural phosphoproteins of mammalian type C viruses. 总被引:14,自引:14,他引:0 下载免费PDF全文
The major phosphoprotein common to woolly monkey sarcoma virus, gibbon ape lymphosarcoma virus, and type C viruses of the lower mammalian species (mouse, rat, cat), with the exception of the endogenous cat virus (RD-114), is the polypeptide of about 12,000 molecular weight. The protein-phosphate bond in this polypeptide of several viruses is of the phosphoserine variety excepting gibbon ape virus, which contains both phosphoserine and phosphothreonine. The primary phosphoprotein of RD-114 virus and the endogenous baboon type C virus, on the other hand, is the polypeptide of about 15,000 molecular weight which contains phosphothreonine as its phosphoamino acid. A second major phosphoprotein of molecular weight of 10,000 is detected only in viruses genetically related to rat species including those derived from the RPL cell line, from Sprague-Dawley rat embryo cells, and the Kirsten mouse sarcoma virus which was recovered from a mouse erythroblastosis virus after in vivo propagation through rat. These phosphorylated polypeptides of molecular weight 15,000, 12,000, or 10,000 are present in the virion structure in several different but nonrandom phosphorylated states. 相似文献
30.