全文获取类型
收费全文 | 2751篇 |
免费 | 331篇 |
国内免费 | 202篇 |
专业分类
3284篇 |
出版年
2024年 | 27篇 |
2023年 | 70篇 |
2022年 | 70篇 |
2021年 | 111篇 |
2020年 | 154篇 |
2019年 | 172篇 |
2018年 | 149篇 |
2017年 | 88篇 |
2016年 | 101篇 |
2015年 | 94篇 |
2014年 | 159篇 |
2013年 | 187篇 |
2012年 | 117篇 |
2011年 | 125篇 |
2010年 | 88篇 |
2009年 | 98篇 |
2008年 | 105篇 |
2007年 | 118篇 |
2006年 | 88篇 |
2005年 | 105篇 |
2004年 | 76篇 |
2003年 | 86篇 |
2002年 | 69篇 |
2001年 | 57篇 |
2000年 | 49篇 |
1999年 | 64篇 |
1998年 | 37篇 |
1997年 | 56篇 |
1996年 | 32篇 |
1995年 | 61篇 |
1994年 | 59篇 |
1993年 | 51篇 |
1992年 | 39篇 |
1991年 | 37篇 |
1990年 | 30篇 |
1989年 | 41篇 |
1988年 | 24篇 |
1987年 | 14篇 |
1986年 | 17篇 |
1985年 | 16篇 |
1984年 | 34篇 |
1983年 | 28篇 |
1982年 | 28篇 |
1981年 | 20篇 |
1980年 | 16篇 |
1979年 | 5篇 |
1978年 | 1篇 |
1977年 | 3篇 |
1976年 | 6篇 |
1974年 | 2篇 |
排序方式: 共有3284条查询结果,搜索用时 15 毫秒
11.
Mortimer M. Civan Stephen R. Williams David G. Gadian Enrique Rozengurt 《The Journal of membrane biology》1986,94(1):55-64
Summary Swiss mouse 3T3 cells grown on microcarrier beads were superfused with electrolyte solution during continuous NMR analysis. Conventional31P and19F probes of intracellular pH (pH
c
) were found to be impracticable. Cells were therefore superfused with 1 to 4mm 2-deoxyglucose, producing a large intracellular, pH-sensitive signal of 2-deoxyglucose phosphate (2DGP). The intracellular incorporation of 2DGP inhibited the Embden-Meyerhof pathway. However, intracellular ATP was at least in part retained and the cellular responsivity to changes in extracellular ionic composition and to the application of growth factors proved intact. Transient replacement of external Na+ with choline or K+ reversibly acidified the intracellular fluids. Quiescent cells and mitogenically stimulated cells displayed the same dependence of shifts in pH
c
on external Na+ concentration (c
Na
o
). pH
c
also depended on intracellular Na+ concentration (c
Na
o
). Increasingc
Na
c
by withdrawing external K+ (thereby inhibiting the Na,K-pump) caused reversible intracellular acidification; subsequently reducingc
Na
o
produced a larger acid shift in pH
c
than with external K+ present. Comparison of separate preparations indicated that pH
c
was higher in stimulated than in quiescent cells. Transient administration of mitogens also reversibly alkalinized quiescent cells studied continuously. This study documents the feasibility of monitoring pH
c
of Swiss mouse 3T3 cells using31P NMR analysis of 2DGP. The results support the concept of a Na/H antiport operative in these cells, both in quiescence and after mitogenic stimulation. The data document by an independent technique that cytoplasmic alkalinization is an early event in mitogenesis, and that full activity of the Embden-Meyerhof pathway is not required for the expression of this event. 相似文献
12.
D. Landowne 《The Journal of membrane biology》1985,88(2):173-185
Summary Measurements of the changes in birefringence associated with changes in membrane potential were made with internally perfused squid giant axons in low sodium solutions at 0–8°C. The time course of the birefringence changes share many properties of the gating (polarization) currents previously studied in this nerve. Both can be demonstrated as an asymmetry in the response to voltage pulses symmetrical about the resting potential which is not present about a hyperpolarized holding potential. Both have a rapid relaxation, which precedes the sodium permeability change. Both exhibit an initial delay or rising phase. Both are reversibly blocked by perfusion with 30mm colchicine; neither are altered by changes on sodium concentrations or 300nm tetrodotoxin. The birefringence response has a decrease in the amplitude of the rapid relaxation associated with the appearance of a slow relaxation. This is similar to the immobilization of fast gating charges which parallels sodium current inactivation.The amplitude of the birefringence and the gating current responses is consistent with a change in the alignment of several hundred peptide bonds per sodium channel. 相似文献
13.
Summary A model has been developed for 5-nitroxide stearate, I(12,3), distribution in human erythrocyte ghosts which accurately predicts ESR spectral alterations observed with increased probe/total lipid (P/L) at 37°C. This spin probe occupies a class of high-affinity, noninteracting sites at low loading. Saturation occurs with increasing probe concentration, and, at higher loading, the probe inserts itself at initially dilute sites to form membranebound clusters of variable size. No low probe remains at high P/L where all I(12,3) clusters in a concentrated phase. This model allows determination of the dilute/clustered probe ratio, and shows that I(12,3) segregates in erythrocytes at what might otherwise be considered low P/L (e.g., 1/359). These findings validate the earlier use of empirical parameters to estimate probe sequestration in biological membranes. 相似文献
14.
Summary Cellular distribution of insulin receptors was studied in fractionated rat liver cell suspensions using 1251-insulin and a visual probe consisting of latex beads covalently linked to insulin (minibeads). Fractionation was done on metrizamide gradients which yielded two cellular fractions. The large cell fraction consisted mostly of hepatocytes and the small cell fraction consisted of 37% endothelial cells as well as Kupffer cells. The magnitude of insulin uptake by the endothelium-rich small cell fraction was at least double that of the uptake by the hepatocyte-rich fraction. The minibead technique demonstrated that in the small cell fraction only endothelial cells, and not Kupffer cells, were responsible for the insulin uptake. Our findings suggest that liver endothelium may be responsible for the uptake of circulating insulin and its transport to hepatocyte. This emphasizes the presence of a tissue-blood barrier in the liver.Abbreviations PRS
phosphate-buffered saline
- SEM
scanning electron microscopy
- TEM
transmission electron microscopy 相似文献
15.
Sexing the human fetus and identification of polyploid nuclei by DNA-DNA in situ hybridisation in interphase nuclei 总被引:1,自引:0,他引:1
J D West C M Gosden J R Gosden K M West Z Davidson C Davidson K H Nicolaides 《Molecular reproduction and development》1989,1(2):129-137
Samples of human adult lymphocytes, fetal lymphocytes, amniotic fluid cells, and chorionic villus cells were sexed independently by cytogenetics and DNA-DNA in situ hybridisation to a tritiated Y probe. For the in situ hybridisation analysis, the presence of Y bodies (hybridisation bodies) in 100 interphase nuclei were scored after autoradiography. In all, 82/83 samples were sexed in this way (one technical failure) and 78/82 were sexed by both in situ hybridisation and cytogenetics. There was complete agreement between the two methods. There was a considerable variation (40-100%) in the percentage of interphase nuclei with a hybridisation body among the male samples, but very few nuclei from female samples showed significant hybridisation. In situ hybridisation could be used to sex the conceptus when males but not females are at risk for various X-linked genetic disorders and may also be useful for detecting 45,X/46,XY mosaicism or polyploid/diploid mosaicism. This would be particularly useful for direct preparations of chorionic villus samples, which often prove difficult to analyse cytogenetically but offer the best means of avoiding maternal contamination. Some interphase nuclei had more than one hybridisation body, and this was most commonly found among amniotic fluid cells. Comparison of sizes of nuclei with one or two hybridisation bodies strongly suggested that most of the amniotic fluid cell nuclei with two hybridisation bodies were tetraploid. 相似文献
16.
The enantioseparation of the sherry aroma components 5-oxo-4-hydroxyhexanoic acid γ-lactone (solerone) and 4,5-dihydroxyhexanoic acid γ-lactone (solerole) is achieved, using Chiraspher (Merck) as the chiral HPLC phase and the optical purity ascertained directly by HRGC with heptakis(3-O-acetyl-2,6-di-O-pentyl)-β-cyclodextrin (Lipodex D) as the chiral stationary phase. The absolute configurations of 4,5-dihydroxyhexanoic acid γ-lactones are assigned by 1H-NMR spectral data of diastereomeric α-methoxy-α-trifluoromethylphenylacetic acid (MTPA) esters, according to Mosher's model. Sensory qualities of the isomers are given. 相似文献
17.
Enantiomers of diastereomeric benzo[a]pyrene (BP) diol-epoxides, r-7,t-8-dihydroxy-t-9,10-epoxy-7,8,9,10-tetrahydro-BP (BP 7,8-diol-anti-9,10-epoxide), r-7,t-8-dihydroxy-c-9,10-epoxy-7,8,9,10-tetrahydro-BP (BP 7,8-diol-syn-9,10-epoxide), r-9,t-10-dihydroxy-t-7,8-epoxy-7,8,9,10-tetrahydro-BP (BP 9,10-diol-anti-7,8-epoxide), and several 7,8,9,10-tetrahydroxy-7,8,9,10-tetrahydrobenzo[a]pyrenes (BP tetrols) were resolved by high-performance liquid chromatography (HPLC) using columns packed with either (R)-N-(3,5-dinitrobenzoyl)phenylglycine[(R)-DNBPG] or (S)-N-(3,5-dinitrobenzoyl)leucine [(S)-DNBL], which is either ionically or covalently bonded to gamma-aminopropylsilanized silica. Resolution of enantiomers was confirmed by ultraviolet-visible absorption and circular dichroism spectral analyses. Resolved enantiomers of BP diol-epoxides were each hydrolyzed in acidic solution to a pair of diastereomeric tetrols which were separated by reversed-phase HPLC. Absolute stereochemistries of enantiomeric diol-epoxides were deduced by the absolute configuration of their hydrolysis products. 相似文献
18.
Determination of the pressure in the water-conducting vessels of intactNicotiana rustica L. plants showed that the pressure probe technique gave less-negative values than the Scholander-bomb method. Even though
absolute values of the order of −0.1 MPa could be directly recorded in the xylem by means of the pressure probe, pressures
between zero and atmospheric were also frequently found. The data obtained by the pressure probe for excised leaves showed
that the Scholander bomb apparently did not read the actual tension in the xylem vessles ofNicotiana plants. The possibility that the pressure probe gave false readings was excluded by several experimental controls. In addition,
cavitation and leaks either during the insertion of the microcapillary of the pressure probe, or else during the measurements
were easily recognized when they occurred because of the sudden increase of the absolute xylem tension to that of water vapour
or to atmospheric, respectively. Tension values of the same order could also be measured by means of the pressure probe in
the xylem vessels of pieces of stem cut from leaves and roots under water and clamped at both ends. The magnitude of the absolute
tension depended on the osmolarity of the bathing solution which was adjusted by addition of appropriate concentrations of
polyethylene glycol. Partial and uniform pressurisation of plant tissues or organs, or of entire plants (by means of the Scholander
bomb or of a hyperbaric chamber, respectively) and simultaneous recording of the xylem tension using the pressure probe showed
that a 1∶1 response in xylem pressure only occurred under a few circumstances. A 1∶1 response required that the xylem vessels
were in direct contact with an external water reservoir and/or that the tissue was (pre-)infiltrated with water. Corresponding
pressure-probe measurements in isolated vascular bundles ofPlantago major L. orP. lanceolata L. plants attached to a Hepp-type osmometer indicated that the magnitude of the tension in the xylem vessels was determined
by the external osmotic pressure of the reservoir. These and other experiments, as well as analysis of the data using classical
thermodynamics, indicated that the turgor and the internal osmotic pressure of the accessory cells along the xylem vessels
play an important role in the maintenance of a constant xylem tension. This conclusion is consistent with the cohesion theory.
In agreement with the literature (P.E. Weatherley, 1976, Philos. Trans. R. Soc. London Ser. B23, 435–444; 1982, Encyclopedia of plant physiology, vol. 12B, 79-109), it was found that the tension in the xylem of intact
plants under normal and elevated ambient pressure (as measured with the pressure probe) under quasi-stationary conditions
was independent of the transpiration rate over a large range, indicating that the conductance of the flow path must be flow-dependent. 相似文献
19.
K. Anamthawat-Jónsson T. Schwarzacher A. R. Leitch M. D. Bennett J. S. Heslop-Harrison 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1990,79(6):721-728
Summary Labelled total genomic DNA was used as a probe in combination with blocking DNA to discriminate between taxonomically closely related species in the genera Hordeum and Secale. Discrimination was possible both by Southern hybridization to size-fractionated restriction enzyme digests of genomic DNA and by in situ hybridization to chromosome preparations. To distinguish between two species (e.g. H. vulgare and H. bulbosum), genomic DNA from one species was used as the labelled probe, while unlabelled DNA from the other species was applied at a much higher concentration as a block. The blocking DNA presumably hybridized to sequences in common between the block and the labelled probe, and between the block and DNA sequences on the membrane or chromosomes in situ. If so, mainly species-specific sequences would remain as sites for probe hybridization. These species-specific sequences are dispersed and represent a substantial proportion of the genome (unlike many cloned, species-specific sequences). Consequently, rapid nonradioactive methods detected probe hybridization sites satisfactorily. The method was able to confirm the parentage of hybrid plants. It has potentially wide application in plant breeding for the detection of alien DNA transfer, and it can be easily adapted to many species. 相似文献
20.
Ellen J. Lehning Renu Doshi Norman Isaksson Peter K. Stys Richard M. LoPachin Jr. 《Journal of neurochemistry》1996,66(2):493-500
Abstract: To investigate the route of axonal Ca2+ entry during anoxia, electron probe x-ray microanalysis was used to measure elemental composition of anoxic tibial nerve myelinated axons after in vitro experimental procedures that modify transaxolemmal Na+ and Ca2+ movements. Perfusion of nerve segments with zero-Na+/Li+-substituted medium and Na+ channel blockade by tetrodotoxin (1 µM) prevented anoxia-induced increases in Na and Ca concentrations of axoplasm and mitochondria. Incubation with a zero-Ca2+/EGTA perfusate impeded axonal and mitochondrial Ca accumulation during anoxia but did not affect characteristic Na and K responses. Inhibition of Na+-Ca2+ exchange with bepridil (50 µM) reduced significantly the Ca content of anoxic axons although mitochondrial Ca remained at anoxic levels. Nifedipine (10 µM), an L-type Ca2+ channel blocker, did not alter anoxia-induced changes in axonal Na, Ca, and K. Exposure of normoxic control nerves to tetrodotoxin, bepridil, or nifedipine did not affect axonal elemental composition, whereas both zero-Ca2+ and zero-Na+ solutions altered normal elemental content characteristically and significantly. The findings of this study suggest that during anoxia, Na+ enters axons via voltage-gated Na+ channels and that subsequent increases in axoplasmic Na+ are coupled functionally to extraaxonal Ca2+ import. Intracellular Na+-dependent, extraaxonal Ca2+ entry is consistent with reverse operation of the axolemmal Na+-Ca2+ exchanger, and we suggest that this mode of Ca2+ influx plays a general role in peripheral nerve axon injury. 相似文献