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61.
(1) Exposure of phospholipids at the outer surface of activated and control platelets was studied by incubation with a mixture of phospholipase A2 from Naja naja and bee venom, solely or in combination with sphingomyelinase from Staphylococcus aureus, using conditions under which cell lysis remained below 10%. (2) Incubation with phospholipase A2 alone revealed a markedly increased susceptibility of the phospholipids in platelets activated by a mixture of collagen plus thrombin, by the SH-oxydizing compound diamide, or by calcium ionophore A23187, as compared to control platelets or platelets activated separately by collagen or thrombin. (3) Collagen plus thrombin, diamide, and ionophore treated platelets revealed an increased exposure of phosphatidylserine at the outer surface accompanied by a decreased exposure of sphingomyelin, as could be concluded from incubations with a combination of phospholipase A2 and sphingomyelinase. These alterations were much less apparent in platelets activated either by thrombin or by collagen alone. (4) The increased exposure of phosphatidylserine in activated platelets is accompanied by an increased ability of the platelets to enhance the conversion of prothrombin to thrombin by coagulation factor Xa, in the presence of factor Va and calcium. (5) It is concluded that the altered orientation of the phospholipids in the plasma membrane of platelets activated by collagen plus thrombin, by diamide, or by calcium ionophore, is the result of a transbilayer movement. Moreover, the increased exposure of phosphatidylserine in platelets stimulated by the combined action of collagen and thrombin might be of considerable importance for the hemostatic process. 相似文献
62.
Forskolin, an activator of adenylate cyclase, stimulates adrenocorticotropin (ACTH) release and increases proopiomelanocortin mRNA levels in anterior pituitary cells by enhancing cyclic AMP (cAMP)-dependent protein kinase activity. The phorbol ester phorbol 12-myristate 13-acetate (PMA) evokes these same responses from anterior pituitary cells by activating protein kinase C. Both protein kinases most likely induce their cellular effects by catalyzing the phosphorylation of specific proteins. To elucidate the mechanisms by which cAMP-dependent protein kinase and protein kinase C promote ACTH secretion and synthesis, the phosphoproteins regulated by forskolin and PMA were identified in the cell line AtT-20, which consists of a homogeneous population of corticotrophs. Phosphoproteins were analyzed in different subcellular fractions by two-dimensional polyacrylamide gel electrophoresis and autoradiography. Forskolin increased phosphate incorporation into two proteins in the cytoplasmic fraction of 24 kilodaltons (kd) (pI 6.8) and 40 kd (pI 5.8), two proteins in the plasma membrane fraction of 32 kd (pI 8.3) and 60 kd (pI 8), and one protein in the nuclear fraction of 20 kd (pI 8.7). Insertion of the inhibitor of cAMP-dependent protein kinase into the AtT-20 cells, using a liposome technique, blocked the rise in phosphate incorporation induced by forskolin. PMA also stimulated phosphate incorporation into proteins in AtT-20 cells. PMA increased the phosphorylation of three cytoplasmic proteins of 25 kd (pI 7.6), 40 kd (pI 5.8), and 40 kd (pI 8.1) as well as two membrane proteins of 32 kd (pI 8.3) and 60 kd (pI 8) and one nuclear protein of 20 kd (pI 6.3).(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
63.
Versieck Jacques Vanballenberghe Lidia Wittoek Ann Vermeir Gerda Vandecasteele Carlo 《Biological trace element research》1990,(1):683-689
A method is described for the determination of mercury in human blood serum and packed blood cells employing neutron activation
analysis. Great attention was devoted to the collection and manipulation of the samples. The accuracy and precision of the
method were tested by analyzing biological reference materials and by comparing the concentrations measured in a number of
serum samples to those obtained by another, independent technique (cold vapor atomic absorption spectrometry) in the same
samples. The article reports the levels measured in blood serum and packed blood cells samples from 15 adult volunteers, as
well as the figures determined in a “second-generation” biological reference material (freeze-dried human serum), prepared
and conditioned at the University of Ghent. 相似文献
64.
Krishnan S. S. McNeill K. G. Mernagh J. R. Harrison J. E. 《Biological trace element research》1990,(1):415-421
Two new facilities for in vivo activation analysis of patients have been designed, developed, and constructed at Toronto General
Hospital. One of these is for the determination of body calcium for the diagnosis of osteoporosis and other diseases associated
with bone loss. The other is for the measurement of total body nitrogen for the determination of protein status.
These facilities replace old university facilities and take into account the comfort and management of patients. In addition,
in the case of the calcium facility, the precision of the measurements has been improved because of larger detector volume
and increased neutron source strength. Both the facilities are now in routine hospital clinical use. 相似文献
65.
Edward J. B. Beeley P. A. Bennett L. G. I. Poland J. S. 《Biological trace element research》1990,(1):53-61
A microcomputer-controlled irradiation and measurement system and a microprocessor-controlled sample changer have been installed
at the SLOWPOKE-2 Facility at the Royal Military College of Canada (RMC). These systems can provide the gamut of instrumental
neutron activation analysis (INAA) techniques for the analyst. Custom software has been created for system control, data acquisition,
and off-line spectral analysis using programs that incorporate Gaussian peak-fitting methods of analysis. The design and use
of the equipment is discussed, and the performance is illustrated with results obtained from the analysis of marine sediment
and biological reference materials. 相似文献
66.
The green alga Chlamydomonas reinhardtii is a facultative heterotroph and, when cultured in the presence of acetate, will synthesize chlorophyll (Chl) and photosystem (PS) components in the dark. Analysis of the thylakoid membrane composition and function in dark grown C. reinhardtii revealed that photochemically competent PS II complexes were synthesized and assembled in the thylakoid membrane. These PS II centers were impaired in the electron-transport reaction from the primary-quinone electron acceptor, QA, to the secondary-quinone electron acceptor, QB (QB-nonreducing centers). Both complements of the PS II Chl a–b light harvesting antenna (LHC II-inner and LHC II-peripheral) were synthesized and assembled in the thylakoid membrane of dark grown C. reinhardtii cells. However, the LHC II-peripheral was energetically uncoupled from the PS II reaction center. Thus, PS II units in dark grown cells had a -type Chl antenna size with only 130 Chl (a and b) molecules (by definition, PS II units lack LHC II-peripheral). Illumination of dark grown C. reinhardtii caused pronounced changes in the organization and function of PS II. With a half-time of about 30 min, PS II centers were converted froma QB-nonreducing form in the dark, to a QB-reducing form in the light. Concomitant with this change, PS II units were energetically coupled with the LHC II-peripheral complement in the thylakoid membrane and were converted to a PS II form. The functional antenna of the latter contained more than 250 Chl(a+b) molecules. The results are discussed in terms of a light-dependent activation of the QA-QB electron-transfer reaction which is followed by association of the PS II unit with a LHC II-peripheral antenna and by inclusion of the mature form of PS II (PS II) in the membrane of the grana partition region.Abbreviations Chl
chlorophyll
- PS
photosystem
- QA
primary quinone electron acceptor of PS II
- QB
secondary quinone electron acceptor of PS II
- LHC
light harvesting complex
- F0
non-variable fluorescence yield
- Fplf
intermediate fluorescence yield plateau leyel
- Fmax
maximum fluorescence yield
- Fi
initial fluorescence yield increase from F0 to Fpl (Fpl–F0)
- Fv
total variable fluorescence yield (Fm–F0)
- DCMU
dichlorophenyl-dimethylurea 相似文献
67.
Bovine kidneys were found to contain about 78 ppm Zn and 0.78 ppm Cd. Approximately 45% of Zn and 60% of Cd were present in
the cytosol fraction. More than 95% of these two metals were bound to macromolecules. Both Zn- and Cd-protein complexes were
observed to be stable between pH 7 and 10.5. Separation and characterization of these proteins were carried out using several
chromatographic and electrophoretic techniques in conjunction with instrumental neutron activation analysis (INAA). The results
showed the presence of at least four Zn-binding proteins with mol wt>300,000, 260,000, 89,000, and 27,000 and at least three
Cd-binding proteins of mol wt>300,000, 32,000, and 13,000. 相似文献
68.
An epithermal neutron activation method is used to determine the concentration of mineral elements in human dental enamel.
A large number (252) of samples from ancient and modern origins are analyzed. The analytical results are mathematically processed
using a statistical multivariant method. This allows to differentiate deciduous from permanent teeth and decayed from sound
enamel. It is also possible to distinguish the teeth coming from two different necropoles. The origin and the localization
of determined elements in the mineralized part, or in the aqueous-organic part, of enamel is suggested. Their role, as witnessed
in the physiopathological phenomena of dental enamel, is discussed. 相似文献
69.
The major developments in the field of nuclear activation analysis, from 1936 to 1989, are discussed. The developments are
grouped into five consecutive time periods. The impact of various scientists on the development of the field in the first
35 years is also discussed. 相似文献
70.
By indirect immunofluorescence, using rabbit anti-heparin-binding placental protein (HBPP) antiserum, we studied HBPP expression by physiologically and non-physiologically (microsurgically) activated hamster gametes. Whereas mature gametes (sperm, metaphase II oocytes) were negative, in vivo conceived preimplantation embryos, from pronuclear to two- and four-cell stages, were HBPP positive. No HBPP was demonstrated in the zona pellucida, but HBPP-dependent immunofluorescence was localized in the perivitelline space. Oocytes incubated with hyaluronidase demonstrated variable responses from negative to positive. (Diluent or sperm) microinjected oocytes were all activated and HBPP positive within 4 h after stimulation. Thus neither activation by microinjection nor HBPP expression required paternal gametes. These kinetics suggest that HBPP may be a cortical granule secretogogue which can be applied to monitor oocyte responses during in vitro manipulations. 相似文献