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21.
22.
Alfonso Pompella Caterina Cambiaggi Silvia Dominici Aldo Paolicchi Roberto Tongiani Mario Comporti 《Histochemistry and cell biology》1996,105(3):173-178
Confocal laser scanning fluorescence microscopy coupled to image analysis was employed in order to develop and evaluate procedures for the appraisal at the single-cell level of: (1) protein-bound 4-hydroxynonenal, the specific product of membrane peroxidation (by means of immunocytochemistry with biotin-avidin revelation); (2) protein oxidation (by reaction of protein carbonyls with 2,4-dinitrophenyl-hydrazine followed by immunocytochemistry of dinitrophenyl moieties); and (3) cellular protein thiols (by direct alkylation of sulfhydryl groups with thiol-specific fluorescent reagents possessing different cell permeabilities). The procedures proved able to reveal the subcellular distribution of cytochemical parameters useful as indices of oxidative stress conditions, and may allow redox phenotyping of isolated cells, which would provide an efficient tool in selected experimental models. 相似文献
23.
Alfonso Pompella Aldo Paolicchi Silvia Dominici Mario Comporti Roberto Tongiani 《Histochemistry and cell biology》1996,106(3):275-282
A number of studies indicate that cell proliferation can be modulated by changes in the redox balance of (soluble and protein)
cellular thiols. Free radical processes, including lipid peroxidation (LPO), can affect such a balance, and a role for LPO
in multistage carcinogenesis has been envisaged. The present study was aimed to assess the relationships between the protein
thiol redox status and the LPO process in chemically induced preneoplastic tissue. The Solt-Farber's initiation-promotion
model of chemical carcinogenesis in the rat liver was used. In fresh cryostat sections, preneoplastic lesions were identified
by the reexpression of γ-glutamyltranspeptidase (GGT) activity. In serial sections, different classes of protein thiols were
stained; in additional sections, LPO was elicited by various prooxidant mixtures and determined thereafter by the hydroxynaphthoic
hydrazide-Fast Blue B procedure. The incubation of sections in the presence of chelated iron plus substrates for GGT activity
leads to the development of LPO in selected section areas closely corresponding to GGT-positive lesions, indicating the ability
of GGT activity to initiate LPO. Protein-reactive thiols, as well as total protein sulfur, were decreased by 20–25% in cells
belonging to GGT-positive preneoplastic nodules, suggesting the occurrence of oxidative conditions in vivo. The incubation
of additional adjacent sections with the prooxidant mixture H2O2 plus iron(II), in order to induce the complete oxidation of lipid present in the section, showed a decreased basal concentration
of oxidizable lipid substrate in GGT-rich areas. The decreased levels of both protein thiols and lipid-oxidizable substrate
in GGT-positive nodules suggest that the observed GGT-dependent path-way of LPO initiation can be chronically operative in
vivo during early stages of chemical carcinogenesis, in cells expressing GGT as part of their transformed phenotype. 相似文献
24.
A Mutation in the D-de Loop of D1 Modifies the Stability of the S2QA- and S2QB- States in Photosystem II 总被引:1,自引:0,他引:1 下载免费PDF全文
Maenpaa P Miranda T Tyystjarvi E Tyystjarvi T Govindjee Ducruet JM Etienne AL Kirilovsky D 《Plant physiology》1995,107(1):187-197
Photosystem II electron transfer, charge stabilization, and photoinhibition were studied in three site-specific mutants of the D1 polypeptide of Synechocystis PCC 6803: E243K, E229D, and CA1 (deletion of three glutamates 242-244 and a substitution, glutamine-241 to histidine). The phenotypes of the E229D and E243K mutants were similar to that of the control strain (AR) in all of the studied aspects. The characteristics of CA1 were very different. Formate, which inhibits the QA- to QB- reaction, was severalfold less effective in CA1 than in AR. The S2QA- and S2QB- states were stabilized in CA1. It was previously shown that the electron transfer between QA- and QB was modified in CA1 (P Maenpaa, T. Kallio, P. Mulo, G. Salih, E.-M. Aro, E. Tyystjarvi, C. Jansson [1993] Plant Mol Biol 22: 1-12). A change in the redox potential of the QA/QA- couple, which renders the reoxidation of QA- by back or forward reactions more difficult, could explain the phenotype of CA1. Although the rates of photoinhibition measured as inhibition of oxygen evolution, Chl fluorescence quenching, and decrease of thermoluminescence B and Q bands were similar in AR and CA1, the CA1 strain more quickly reached a state from which the cells were unable to recover their activity. The results described in this paper suggest that a modification in the structure of the D-de loop of D1 could influence the properties of the couple QA/QA- in D2 and the mechanism of recovery from photoinhibition. 相似文献
25.
Thermoluminescence experiments have been carried out to study the effect of a transmembrane proton gradient on the recombination properties of the S2 and S3 states of the oxygen evolving complex with QA
- and QB
-, the reduced electron acceptors of Photosystem II. We first determined the properties of the S2QA
- (Q band), S2QB
- and S3QB
- (B bands) recombinations in the pH range 5.5 to 9.0, using uncoupled thylakoids. The, a proton gradient was created in the dark, using the ATP-hydrolase function of ATPases, in coupled unfrozen thylakoids. A shift towards low temperature of both Q and B bands was observed to increase with the magnitude of the proton gradient measured by the fluorescence quenching of 9-aminoacridine. This downshift was larger for S3QB
- than for S2QB
- and it was suppressed by nigericin, but not by valinomycin. Similar results were obtained when a proton gradient was formed by photosystem I photochemistry. When Photosystem II electron transfer was induced by a flash sequence, the reduction of the plastoquinone pool also contributed to the downshift in the absence of an electron acceptor. In leaves submitted to a flash sequence above 0°C, a downshift was also observed, which was supressed by nigericin infiltration. Thus, thermoluminescence provides direct evidence on the enhancing effect of lumen acidification on the S3S2 and S2S1 reverse-transitions. Both reduction of the plastoquinone pool and lumen acidification induce a shift of the Q and B bands to lower temperature, with a predominance of lumen acidification in non-freezing, moderate light conditions.Abbreviations 9-AA
9-aminoacridine
- EA
activation energy
- F0
constant fluorescence level
- FM
maximum fluorescence, when all PS-II centers are closed
- FV
variable fluorescence (FM–F0)
- PS I, PS II
Photosystem I, photosystem II
- PQ
plastoquinone
- TL
thermoluminescence 相似文献
26.
The DNA rearrangement that generates the TRK-T3 oncogene involves a novel gene on chromosome 3 whose product has a potential coiled-coil domain. 总被引:3,自引:1,他引:2 下载免费PDF全文
A Greco C Mariani C Miranda A Lupas S Pagliardini M Pomati M A Pierotti 《Molecular and cellular biology》1995,15(11):6118-6127
Oncogenic rearrangements of the NTRK1 gene (also designated TRKA), encoding one of the receptors for the nerve growth factor, are frequently detected in thyroid carcinomas. Such rearrangements fuse the NTRK1 tyrosine kinase domain to 5'-end sequences belonging to different genes. In previously reported studies we have demonstrated that NTRK1 oncogenic activation involves two genes, TPM3 and TPR, both localized similarly to the receptor tyrosine kinase, on the q arm of chromosome 1. Here we report the characterization of a novel NTRK1-derived thyroid oncogene, named TRK-T3. A cDNA clone, capable of transforming activity, was isolated from a transformant cell line. Sequence analysis revealed that TRK-T3 contains 1,412 nucleotides of NTRK1 preceded by 598 nucleotides belonging to a novel gene that we have named TFG (TRK-fused gene). The TRK-T3 amino acid sequence displays, within the TFG region, a coiled-coil motif that could endow the oncoprotein with the capability to form complexes. The TRK-T3 oncogene encodes a 68-kDa cytoplasmic protein reacting with NTRK1-specific antibodies. By sedimentation gradient experiments the TRK-T3 oncoprotein was shown to form, in vivo, multimeric complexes, most likely trimers or tetramers. The TFG gene is ubiquitously expressed and is located on chromosome 3. The breakpoint producing the TRK-T3 oncogene occurs within exons of both the TFG gene and the NTRK1 gene and produces a chimeric exon that undergoes alternative splicing. Molecular analysis of the NTRK1 rearranged fragments indicated that the chromosomal rearrangement is reciprocal and balanced and involves loss of a few nucleotides of germ line sequences. 相似文献
27.
Caenorhabditis Elegans Mutants Resistant to Inhibitors of Acetylcholinesterase 总被引:4,自引:2,他引:2 下载免费PDF全文
We characterized 18 genes from Caenorhabditis elegans that, when mutated, confer recessive resistance to inhibitors of acetylcholinesterase. These include previously described genes as well as newly identified genes; they encode essential as well as nonessential functions. In the absence of acetylcholinesterase inhibitors, the different mutants display a wide range of behavioral deficits, from mild uncoordination to almost complete paralysis. Measurements of acetylcholine levels in these mutants suggest that some of the genes are involved in presynaptic functions. 相似文献
28.
Alfonso Calvo Luis M. Pastor Ramon Horn Jacinto Pallares 《The Histochemical journal》1995,27(9):670-680
Summary The glycoconjugates of hamster epididymis were investigated with conventional and lectin histochemistry. A zone of the caput
epididymis, with particular histochemical characteristics, has been differentiated. β-Elimination in combination with lectins
was used to establish the presence and distribution of N- and O-linked glycoconjugates. The epithelium, spermatozoa and the
intertubular matrix were rich in glycoconjugates. The Golgi apparatus and stereocilia of the principal cells were intensely
positive with HPA, PNA and SBA lectins. β-limination indicated that these cells contained abundant O-linked glycoconjugates.
Apical and clear cells presented a common lectin affinity; their reactivities towards WGA and UEA-I were very positive. These
cells probably contain abundant N-glycoconjugates. The spermatozoa were stained by periodic acid-Schiff (PAS) and by all the
lectins (especially in the acrosome), except by those with an affinity for α-l-fucosyl residues; the most intense reaction was found with HPA, WGA, PNA and SBA. Changes in the sperm lectin binding along
the ductus were observed: sperm flagellum abruptly acquired WGA and PNA labelling from the posterior caput, and HPA reactivity
was negative only in the zone between the caput and the corpus. 相似文献
29.
The cattle of Doñana (139 individuals in four social groups in 1989) have lived under free-ranging conditions for centuries. Their ranging behaviour was analysed during a three-year period. A total of 17,603 locations corresponding to 247 different animals allowed both for the estimation of global and seasonal home ranges of individuals and social groups and for the comparison of movement patterns. Cattle ranging behaviour was not affected by human interference, and was shown to be regulated by a complex interaction of environment, individual and social factors. Habitat structure and seasonal fluctuations in abundance and distribution of resources determined general patterns of ranging behaviour: the greater the concentration of resources, the smaller the home ranges of individuals and social groups. These patterns were modified at an individual level by the sex of the animal and its reproductive status if male. Social influences on ranging behaviour were important because these implied the segregation of home ranges among dominant bulls and among social groups. As a result, there was a great variability in space use and home-range behaviour. 相似文献
30.