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51.
van der Pol JJ Machnik M Biselli M Portela-Klein T de Gooijer CD Tramper J Wandrey C 《Cytotechnology》1997,24(1):19-30
The monoclonal-antibody production of an immobilized hybridoma cell line cultivated in a fluidized-bed reactor was monitored
on-line for nearly 900 h. The monoclonal antibody concentration was determined by an immuno affinity-chromatography method
(ABICAP). Antibodies directed against the product, e.g. IgG, were immobilized on a micro-porous gel and packed in small columns.
After all IgG present in the sample was bound to the immobilized antibodies, unbound proteins were removed by rinsing the
column. Elution of the bound antibodies followed and the antibodies were determined by fluorescence. The analytical procedure
was automated with a robotic device to enable on-line measurements. The correlation between the on-line determined data and
antibody concentrations measured by HPLC was linear.
A sampling system was constructed, which was based on a pneumatically actuated in-line membrane valve integrated into the
circulation loop of the reactor. Separation of the cells from the sample stream was achieved by a depth filter made of glass-fibre,
situated outside the reactor. Rapid obstruction of the filter by cells or cell debris and contamination of the sample system
was avoided by intermittent rinsing of the sample system with a chemical solution. The intermittent rinsing of the filter,
which had a surface of 4.8 cm2, resulted in an operational capacity of up to 40 samples (1.0 l total sample volume). Both the sampling system and the analytical
device functioned without failure during this long-term culture.
The culture temperature was varied between 34 and 40 °C. Raising the temperature from 34 up to 37 °C resulted in a simultaneous
increase of growth and specific antibody production rate. Specific metabolic rates of glucose, lactate, glutamine and ammonium
stayed constant in this temperature range. A further enhancement of temperature up to 40 °C had a negative effect on the growth
rate, whereas the specific monoclonal antibody production rate showed a small increase. The other specific metabolic rates
also increased in the temperature range between 38 to 40 °C.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
52.
Mikkel Nissum Jens-Jakob Karlsson Jens Ulstrup Palle Waage Jensen G. Smulevich 《Journal of biological inorganic chemistry》1997,2(3):302-307
Di-heme Pseudomonas stutzeri cytochrome c
4 has been characterized by electronic absorption and resonance Raman spectroscopies in the ferric and ferrous forms at pH 7.5
and at room temperature. The data indicate that the two hemes are inequivalent. It is proposed that the N-terminal contains
a more relaxed heme as a consequence of the relative orientation of the methionine and histidine ligands with respect to the
N-Fe-N directions of the heme plane. This causes a weakening of the Fe-S bond with concomitant partial dissociation of the
methionine and the formation of an Fe-aquo bond. Heme group relaxation is further accompanied by less distortion of the heme
group than that associated with cytochrome c, expansion of the "core" and a negative shift of the redox potential.
Received: 17 December 1996 / Accepted: 6 March 1997 相似文献
53.
Abstract: Activation of immediate early gene expression is a key event in stress-induced neuronal cell injury. To study whether changes in cytoplasmic calcium activity are necessary to activate neuronal immediate early gene expression, endoplasmic reticulum (ER) calcium stores of primary neurons were depleted by exposing cells to thapsigargin (Tg), an irreversible inhibitor of ER Ca2+ -ATPase. Tg-induced rise in [Ca2+ ]i and the effect of loading neurons with the cell-permeable calcium chelator BAPTA-AM on this increase in [Ca2+ ]i were measured in fura-2-loaded cells by fluorescence microscopy. Changes in c- fos mRNA levels were evaluated by quantitative PCR. Tg treatment of neurons produced a pronounced rise in c- fos mRNA levels (∼10-fold more than DMSO) which peaked at 1 h after exposure. The Tg-induced rise in c- fos mRNA content was unchanged (hippocampal neurons) or even increased further (cortical neurons) by preloading cells with BAPTA before incubation with Tg. It is concluded that in neuronal cells an increase in cytoplasmic calcium activity is not a prerequisite for a rise in mRNA levels of c- fos . Thus, stress-induced changes in mRNA levels of immediate early genes of neurons may also result from disturbances in ER calcium homeostasis and not necessarily by an overload of cells with calcium ions. The results of the present series of experiments cast further doubt on the widely accepted hypothesis that the stress-induced cytoplasmic overload of neurons with calcium ions is the primary event triggering cell injury. 相似文献
54.
Molecular mechanisms of endotoxin activity 总被引:20,自引:0,他引:20
Jens Schletter Holger Heine Artur J. Ulmer Ernst T. Rietschel 《Archives of microbiology》1995,164(6):383-389
Endotoxin (lipopolysaccharide, LPS), a constitutent of the outer membrane of the cell wall of gramnegative bacteria, exerts
a wide variety of biological effects in humans. This review focuses on the molecular mechanisms underlying these activities
and discusses structure-function relationships of the endotoxin molecule, its interaction with humoral and cellular receptors
involved in cell activation, and transmembrane and intra-cellular signal transduction pathways. 相似文献
55.
Wolf-Dietrich Hard Jens M. Warnecke Roland K. Hartmann 《Molecular biology reports》1995,22(2-3):161-169
Modification interference is a powerful method to identify important functional groups in RNA molecules. We review here recent developments of techniques to screen for chemical modifications that interfere with (i) binding of(pre-)tRNA to bacterial RNase P RNA or (ii) pre-tRNA cleavage by this ribozyme. For example, two studies have analyzed positions at which a substitution of sulfur for thepro-Rp oxygen affects tRNA binding [1] or catalysis [2]. The results emphasize the functional key role of a central core element present in all known RNase P RNA subunits. The four sulfur substitutions identified in one study [2] to inhibit the catalytic step also interfered with binding of tRNA toE. coli RNase P RNA [1]. This suggests that losses in binding energy due to the modification at these positions affect the enzyme-substrate and the enzyme-transition state complex. In addition, the two studies have revealed, for the first time, sites of direct metal ion coordination in RNase P RNA. The potentials, limitations and interpretational ambiguities of modification interference experiments as well as factors influencing their outcome are discussed.Abbreviations nt
nucleotide(s)
- PAGE
polyacrylamide gel electrophoresis 相似文献
56.
Jens Meyer Norbert Elsner 《Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology》1995,176(4):563-573
1. | Up to 9 kHz, the tympanal membrane of the grasshopper Chorthippus biguttulus responds with equal sensitivity at the attachment sites of the low and the high-frequency receptors; at the latter site it is also particularly sensitive between 10 and 20 kHz. |
2. | The frequency spectra of the songs of both sexes exhibit maxima at 7–8 kHz, to which the membrane is well matched. In the high-frequency region, where the male songs have a peak at 30 kHz, there is no corresponding maximum in the membrane oscillation. |
3. | Because the tympanal membrane is immediately adjacent to air sacs in the tracheal system, it is deflected inward and outward by as much as 80 m during the respiratory cycle. |
4. | Measurements by laser vibrometry show that acoustically induced membrane oscillations are attenuated severely due to the respiratory displacement of the membrane for frequencies up to 10–12 kHz. By contrast, at higher frequencies the membrane sensitivity is doubled or tripled. |
5. | As a result of these membrane effects, the discharge in the tympanal nerve was profoundly reduced in the low-frequency range, whereas above 11 kHz there was a marked increase. This modulation of auditory sensitivity affects the animals' ability to detect conspecific songs. |
57.
Sulphur-heterotrophic growth exhibited a dual response to the expression of sulphate-assimilating enzymes. The level of ATP-sulphurylase (EC 2.7.7.4) appeared repressed while sulphite reductase (EC 1.8.7.1) and O-acetyl-l-serine sulphhydrylase (EC 4.2.99.8) were derepressed and coordinated in their occurrence. The capability of the cells to reduce adenylylphosphosulphate or 3-phospho adenylylphosphosulphate to cysteine coincided with the activity of sulphite reductase. The expression of these reducing steps lacked correlation with the regulation of ATP-sulphurylase.Abbreviations APS
adenylylphosphosulphate
- MVH
reduced methylviologen
- OAS
O-acetyl-l-serine
- PAPS
3-phospho adenylylphosulphate 相似文献
58.
Alchemilla austriaca is a new species which belongs to the group ofA. demissa, A. frigens, A. longana, A. longiuscula, A. semisecta, andA. sinuata. The holotype specimen as well as leaf and flower details are illustrated (Figs. 1–3). A complete character analysis is given, differences and similarities of allied species are presented in two tables, and the position of the group within the genus is discussed.A. austriaca so far is known only from the Austrian Alps and mainly from the central ranges (distribution map: Fig. 4). Its wet subalpine and alpine habitats are characterized by species lists. 相似文献
59.
An improved timm sulphide silver method for light and electron microscopic localization of heavy metals in biological tissues 总被引:3,自引:0,他引:3
Summary Modifications of the Timm sulphide silver method for the demonstration of heavy metals are described.To improve the structural preservation of the tissues perfusion with a glutaraldehyde fixative is employed before perfusion with the sodium sulphide solution. For the subsequent staining for light and electron microscopy, procedures for plastic embedding, paraffin embedding and cryostat sectioning are presented. Examples from several tissues are shown, including the pituitary, pancreas, intestine, tongue, kidney, testis and brain. The staining of autolytic, postmortal human brain tissue is demonstrated. 相似文献
60.