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71.
Abstract: Previous work has shown that nerve growth factor (NGF) stimulates the phosphorylation of the ribosomal protein S6 in PC12 cells. In this study, we show that S6 kinase activity is also present in purified PC12 cell nuclei. This activity was increased by treatment of the cells with NGF and, to a lesser extent, by treatment with epidermal growth factor. The NGF-stimulated activity was obtained from nuclear extracts and some of its characteristics described. The increase in activity was prevented by treatment of the cells with rapamycin or with wortmannin, and the overall activity could be precipitated by antibodies directed against the p85S6K . These data indicate that p85S6K is the NGF-stimulated S6 kinase in PC12 cell nuclei. The presence of S6 protein in the nucleus of PC12 cells has been confirmed and evidence is presented that suggests that it is identical to a protein called SMP reported some years ago. 相似文献
72.
Masashi Yamada Toshihiko Ikeuchi Saburo Aimoto Dr. Hiroshi Hatanaka 《Neurochemical research》1996,21(7):815-822
PC12h-R cell, a subclone of PC12 cells, exhibited a neuron-like phenotype, including neurite outgrowth and increased acetylcholinesterase
activity, in response to epidermal growth factor (EGF) as well as nerve growth factor (NGF). We examined the mechanism by
which EGF induced the neuronal differentiation in PC12h-R cells. The EGF-induced neuronal differentiation of PC12h-R cells
was not blocked by K252a, whereas that induced by NGF was. EGF induced sustained tyrosine phosphorylation of the EGF receptor
in PC12h-R cells, but not in the parent PC12h cells, which do not show neuronal differentiation in response to EGF. In addition,
the rate of EGF-induced down-regulation of the EGF receptor in PC12h-R cells was decreased compared with that in PC12h cells.
Furthermore, we found that the duration of EGF-induced tyrosine phosphorylation of the EGF receptor in PC12h-R cells was similar
to that of NGF-induced tyrosine phosphorylation of p140
trkA in PC12h cells. The EGF-induced phosphorylation of the EGF receptor in PC12h cells was less sustained than that of p140
trkA by NGF in PC12h cells. These findings suggested that the EGF-induced neuronal differentiation of PC12h-R cells is due to
the sustained activation of the EGF receptor, resulting from the decreased down-regulation of the EGF receptor and that the
duration of the receptor tyrosine kinase activity determines the cellular responses of PC12 cells. We concluded that sustained
activation of the receptor tyrosine kinase induces neuronal differentiation, although transient activation promotes proliferation
of PC12 cells.
Special issue dedicated to Dr. Hans Thoenen. 相似文献
73.
By using monoclonal antibodies directed against discoidin II,we have isolated cDNA clones from axenically grown Ax-2 cells.On cDNA clone (D2) condtained a 1.2-k.b insert encoding theentire discoidin II protein, which is conposed of 257 aminoacid residuces and has a calculated molecular mass of 28,574.The amino acid sequences, determined by Edman degradation ofsix tryptic peptides of discoidin II, were identical to thosededuced from the cDNA sequences. The protein bears no resemblanceto any proteins in the data banks, except that its sequenceis 49% identical with the amino acid sequence of discoidin I.Discoidin II shares with discoidin I both a carbohydratebindingsite and an Arg-Gly-Asp (RGD) sequence, which has been foundin fibronectin in mammalian cells. With the onset of aggregation(8 h of development), a 1.3-kb discoidin II mRNA begins to accumulate.A similar pattern of regulation occurs at the protein level.
1Present address: MRC Laboratory for Molecular Cell Biology,University College London, Gower Street, London, WC1E 6BT UnitedKingdom 相似文献
74.
Fujimoto J Nishigaki M Hori M Ichigo S Morishita S Tamaya T 《Journal of biomedical science》1995,2(2):160-165
The mechanism of known receptor-mediated androgen effects on the endometrial stroma was studied in endometrial fibroblasts derived from human uterus. 17-Estradiol (E) induced the expressions of androgen receptor (AR) mRNA, and predominantly increased the level of testosterone-binding sites (TBS) in uterine endometrial fibroblasts. The effect on the level of dihydrotestosterone-binding sites (DHTBS) was similar but smaller. This result suggests that the AR mRNA expressed might encode TBS, but probably not DHTBS. The TBS level increased by estrogen was down-regulated by testosterone (T) + E, but the AR mRNA expression increased by E was not down-regulated by E + T in the fibroblasts. Although the synthesis rate of AR was slightly increased (p<0.05) by E alone or E + T, the degradation rate of AR was significantly accelerated (p<0.05) by E + T in the fibroblasts. This result suggests that T might stimulate the metabolic rate of TBS, but does not inhibit the synthesis rate of AR mRNA to TBS in endometrial fibroblasts. 相似文献
75.
Light-induced changes of cytosolic pH (pHc) and the plasmalemmapotential (Em) in dark-adapted leaf cells of the aquatic plant,Egeria densa were measured simultaneously with double-barreledpH-sensitive microelectrodes. Upon illumination, pHc increasedtransiently and then decreased to a level that was lower thanthe original value, while the plasmalemma was greatly hyperpolarizedafter an initial small depolarization. DCMU inhibited the light-inducedchanges in both pHc and Em. DCMU acted without directly inhibitingthe electrogenic proton pump in the plasmalemma since a decreasein pHc caused by treatment with butyrate (H+-loading) hyperpolarizedthe plasmalemma in DCMU-pretreated cells. N.N-Dicyclohexylcarbodiimide(DCCD) also inhibited the light-induced changes in both pHcand Em. This result may be explained by direct inhibition ofthe proton pump in the plasmalemma by DCCD since the decreasein pHc caused by butyrate did not induce membrane hyperpolarizationin DCCD-treated leaf cells. Fusicoccin induced membrane hyperpolarizationand slight acidification of the cytosol. DCCD inhibited thefusicoccin-induced changes in both pHc and Em. The mechanismof the light-induced changes in pHc is discussed in relationto activities of the proton pump in the plasmalemma and photosynthesis. (Received January 10, 1994; Accepted June 9, 1994) 相似文献
76.
Naoyuki Miyashita Yoshifumi Kubota Masashi Kimura Masamitsu Nakajima Yoshihito Niki Rinzo Soejima Akira Matsumoto 《Microbiology and immunology》1994,38(11):857-864
The isolation of Chlamydia pneumoniae, especially from elderly persons, is generally not easy. Recently, we succeeded in isolating a chlamydial strain, which was designated KKpn-15, from a 57-year-old man suffering from acute bronchitis. It was compared with well established strains of C. pneumoniae, C. trachomatis and C. psittaci, and its biological properties, such as the morphology of elementary bodies (EBs) and inclusions, and the immunochemistry of EB proteins, were investigated. Based on the results obtained in the present study, it was confirmed that the new chlamydial strain, KKpn-15, is a member of the C. pneumoniae strain and that the organisms of KKpn-15 are useful as an antigen for the serodiagnosis and epidemiology of C. pneumoniae infection. 相似文献
77.
Jianyu Zheng Kohei Irifune Kouji Hirai Masashi Nakata Ryuso Tanaka Hiromichi Morikawa 《Journal of plant research》1994,107(4):365-369
In situ hybridization with a biotin-labeled rice ribosomal DNA (rDNA) probe to the somatic metaphase chromosomes of six species ofPhaseolus andVigna (P. angularis, P. calcaratus, P. coccineus, P. vulgaris, V. sesquipedalis andV. sinensis) was done to determine the sites of rDNA. Hybridization signals were present in the terminal and subterminal chromosome regions
of each of the six species. The number of rDNA sites was two inP. angularis andP. calcaratus, four inP. coccineus andP. vulgaris, and six inV. sesquipedalis andV. sinensis. 相似文献
78.
79.
Keiko Miyajima Mamoru Hirata Toshiaki Yoshida Hiroshi Kosaka Akira Okayama 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1994,654(2)
A method for the determination of δ-aminolevulinic acid in plasma of lead-exposed workers by high-performance liquid chromatography with fluorescence detection of a fluorescent δ-aminolevulinic acid derivative (2-methylidineamino-3,5-diacetyl-4,6-dimethylpropionic acid) was established. The detection limit of δ-aminolevulinic acid in plasma was 0.01 μg/ml at a signal-to-noise ratio of 5:1. A linear correlation was obtained between the amounts of δ-aminolevulinic acid injected from 0.01 to 0.5 μg/ml (r = 0.999). The recovery of 0.05 and 0.1 μg/ml of δ-aminolevulinic acid added to plasma with various concentrations of δ-aminolevulinic acid in plasma ranged from 80.0 to 100.8%. This method, combined with the use of an automatic sampler, should facilitate the routine measurement of δ-aminolevulinic acid in plasma. 相似文献
80.
Inhibition of endoplasmic reticulum (ER)-to-Golgi transport induces relocalization of binding protein (BiP) within the ER to form the BiP bodies. 总被引:11,自引:6,他引:5
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Immunofluorescence staining of yeast cells with anti-binding protein (BiP) antibodies shows uniform staining of the endoplasmic reticulum (ER). We have found that overproduction of Sec12p, an ER membrane protein, causes a change of BiP distribution within the cell. Upon induction of Sec12p by the GAL1 promoter, the staining pattern of BiP turns into bright dots scattering in the cell, whereas the staining of Sec12p remains to be the typical ER figure. Overproduction of other ER membrane proteins, HMG-CoA reductase or Sed4 protein, does not induce such relocalization of BiP. Pulse-chase experiments and electron microscopy have revealed that the overproduction of Sec12p inhibits protein transport from the ER to the Golgi apparatus. When the transport is arrested by one of the sec mutations that block the ER-to-Golgi step at the restrictive temperature, the BiP staining also changes into the punctate pattern. In contrast, the sec mutants that block later or earlier steps of the secretory pathway do not induce such change of BiP localization. These observations indicate that relocalization of BiP is caused by the inhibition of ER-to-Golgi transport. Using immunoelectron microscopy, we have found that the punctate staining is because of the accumulation of BiP in the restricted region of the ER, which we propose to call the "BiP body." This implicates existence of ER subdomains in yeast. A vacuolar protein, proteinase A, appears to colocalize in the BiP body when the ER-to-Golgi transport is blocked, suggesting that the BiP body may have a role as the site of accumulation of cargo molecules before exit from the ER. 相似文献