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91.
Michael Hoppert Timothy J. Mahony Frank Mayer David J. Miller 《Archives of microbiology》1995,163(4):300-306
The hydroxylamine oxidoreductase from Nitrosomonas europaea was prepared to apparent electrophoretic homogeneity. Electron microscopy of negatively stained preparations of the sample revealed an overall diameter of about 8.8 nm of the enzyme particle. The native structure was determined as a tetrahedron-like assembly of identical subunits exhibiting four protein masses.Abbreviations
ESI
Electron spectroscopic imaging
-
HAO
Hydroxylamine oxidoreductase 相似文献
92.
R. Guglielmone 《Cell and tissue research》1995,281(1):163-168
Although the cerebrospinal fluid-contacting neurons of the avian paraventricular organ exhibit considerable amounts of catecholamines, they show no tyrosine hydroxylase immunoreactivity. In the quail embryo, the development of these neurons has been studied using the paraformaldeyde-glutaraldeyde method for the fluorescence-histochemical localization of catecholamines. The timing of the appearance of catecholamine fluorescence in cerebrospinal fluid-contacting neurons and that in catecholamine-containing neurons of the brainstem have been compared. The first neurons displaying catecholamine fluorescence are found within the locus coeruleus and the nucleus subcoeruleus ventralis on the 5.5th day of incubation. Catecholaminergic neuronal groups of the medulla and mesencephalon can be identified by embryonic day 7, and fluorescent cerebrospinal fluid-contacting neurons of the hypothalamic paraventricular organ can be first recognized at the 8th day of incubation. If the catecholamine content of cerebrospinal fluid-contacting neurons that lack tyrosine hydroxylase depends upon an uptake mechanism, it may be significant that, in fluorescence-histochemical preparations, these neurons can be identified 1–3 days later than those in which catecholamines are synthesized and from which catecholamines are released at an earlier developmental stage. Moreover, cerebrospinal fluid-contacting neurons that have previously been shown to be tyrosine-hydroxylase immunoreactive, and that lie at the spinal-medullary junction display a different developmental pattern. By fluorescence histochemistry, they can be detected only by embryonic day 10.5. The chemical, developmental and topographical differences suggest that the catecholamine-containing cerebrospinal fluid-contacting elements of the paraventricular organ and those of the spinal cord represent two different subsets of cerebrospinal fluid-contacting neurons whose respective functional roles remain to be investigated. 相似文献
93.
Small interfering RNA (siRNA) molecules achieve sequence-specific gene silencing through the RNA interference (RNAi) mechanism. Here, live-cell and live-animal bioluminescent imaging (BLI) is used to directly compare luciferase knockdown by unmodified and nuclease-stabilized siRNAs in rapidly (HeLa) and slowly (CCD-1074Sk) dividing cells to reveal the impact of cell division and siRNA nuclease stability on the kinetics of siRNA-mediated gene silencing. Luciferase knockdown using unmodified siRNAs lasts approximately 1 week in HeLa cells and up to 1 month in CCD-1074Sk cells. There is a slight increase in the duration of luciferase knockdown by nuclease-stabilized siRNAs relative to unmodified siRNAs after cationic lipid transfection, but this difference is not observed after electroporation. In BALB/cJ mice, a fourfold increase in maximum luciferase knockdown is observed after hydrodynamic injection (HDI) of nuclease-stabilized siRNAs relative to unmodified siRNAs, yet the overall kinetics of the recovery after knockdown are nearly identical. By using a mathematical model of siRNA-mediated gene silencing, the trends observed in the experimental data can be duplicated by changing model parameters that affect the stability of the siRNAs before they reach the cytosolic compartment. Based on these findings, we hypothesize that the stabilization advantages of nuclease-stabilized siRNAs originate primarily from effects prior to and during internalization before the siRNAs can interact with the intracellular RNAi machinery. 相似文献
94.
95.
Strosznajder Joanna Zambrzycka Agata Kacprzak Maria D. Kopczuk Dorota Strosznajder Robert P. 《Neurochemical research》1999,24(10):1277-1284
The reperfusion of previously ischemic brain is associated with exacerbation of cellular injury. Reperfusion occasionally potentates release of intracellular enzymes, influx of Ca2+, breakdown of membrane phospholipids, accumulation of amyloid precursor protein or amyloid -(like) proteins, and apolipoprotein E. In this study, the effect of reperfusion injury on the activity of cerebral cortex enzymes acting on phosphatidyl [3H] inositol (PI) and [l4C-arachidonoyl] PI was investigated. Moreover the effect of amyloid 25–35 on PI degradation by phospholipase(s) of normoxic brain and subjected to ichemia-reperfussion injury was determined. Brain ischemia in gerbils (Meriones unguiculatus) was induced by ligation of both common carotid arteries for 5 min and then brains were perfused for 15 min, 2 h and 7 days. Statistically significant activation of enzyme(s) involved in phosphatidylinositol degradation in gerbils subjected to ischemia-reperfusion injury was observed. Nearly all gerbils showed a higher activity of cytosolic PI phos-pholipase C (PLC) at 15 min after ischemia. Concomitantly, the significant enhancement of the level of DAG and AA radioactivity at this short reperfusion time confirmed the active PI degradation by phospholipase(s) in cerebral cortex and hippocampus. After a prolonged reperfusion time of 7 days after ischemia, both cytosolic and membrane-bound forms of PI-PLC were activated. The question arises if alteration of membranes by the degradation of phospholipids occurring after an ischemic episode potentates the effect of A on membrane-bound enzymes. A neuro-toxic fragment of amyloid, A 25–35, incubated in the presence of endogenous Ca2+, increased significantly the PI-PLC activity of normoxic brain. In its non-aggregated form, A 25–35 activates PI-PLC but in the aggregated form the enzymatic activity decreased. Thus, A 25–35 exerts a similar effect on the membrane-bound PI-PLC from normoxic brain or subjected to ischemia reperfussion injury. We conclude that the degradation of phosphatidylinositol by cytosolic phosphoinositide-phospholipase C may contribute to the pathophysiology of delayed neuronal death following cerebral ischemia. Thus, a specific inhibitor of this enzyme(s) may offer therapeutic strategies to protect the brain from damage triggered by ischemia. Ischemia-reperfusion injury had no effect on A-evoked alterations of synaptic plasma membrane-bound PI-PLC. 相似文献
96.
Anastasia Nikolopoulou Theodosia Maina Petros Sotiriou Paul Cordopatis Berthold A. Nock 《Journal of peptide science》2006,12(2):124-131
Two somatostatin analogues, [99mTc]Demotide and [99mTc]Demotate 4, were compared with [99mTc]Demotate 1, a previously reported somatostatin receptor subtype 2 (sst2) targeting tracer. Conjugates were prepared by coupling an open‐chain tetraamine chelator to D ‐Phe1 of [Tyr3]‐octreotide or [Tyr3]‐octreotate, respectively, via a p‐benzylaminodiglycolic acid spacer adopting solid‐phase peptide synthesis techniques. Peptide conjugates were collected in a highly pure form after chromatographic purification. Eventually, [99mTc]Demotide and [99mTc]Demotate 4 were obtained in ~1 Ci/µmol specific activity and >96% purity after labeling under alkaline conditions. Demotide and Demotate 4 exhibited similar high binding affinities for the sst2 expressed in AR4‐2J cells with IC50 values 0.16 and 0.10 nM, respectively. The (radio)metallated analogues [99mTc]Demotide and [99mTc]Demotate 4 showed equally high affinities to the sst2 during saturation binding assays in AR4‐2J cell membranes (Kds 0.08 and 0.07 nM, respectively). During incubation at 37 °C with AR4‐2J cells, the radiopeptides internalized effectively via a receptor‐mediated process, with [99mTc]Demotate 4 exhibiting a faster internalization rate than [99mTc]Demotide. After injection in athymic mice bearing sst2‐expressing AR4‐2J tumors, the radiotracers showed high and specific uptake in the tumor (>25%ID/g at 1 h) and in the sst2–positive organs. However, both [99mTc]Demotide and [99mTc]Demotate 4 showed unfavorably higher background activity, especially in the abdomen, in comparison to [99mTc]Demotate 1 and are, therefore, less suited than [99mTc]Demotate 1 for sst2‐targeted tumor imaging in man. Copyright © 2005 European Peptide Society and John Wiley & Sons, Ltd. 相似文献
97.
Calcification process dynamics in coral primary polyps as observed using a calcein incubation method
Yoshikazu Ohno Akira Iguchi Chuya Shinzato Mikako Gushi Mayuri Inoue Atsushi Suzuki Kazuhiko Sakai Takashi Nakamura 《Biochemistry and Biophysics Reports》2017
Calcification processes are largely unknown in scleractinian corals. In this study, live confocal imaging was used to elucidate the spatiotemporal dynamics of the calcification process in aposymbiotic primary polyps of the coral species Acropora digitifera. The fluorophore calcein was used as a calcium deposition marker and a visible indicator of extracellular fluid distribution at the tissue-skeleton interface (subcalicoblastic medium, SCM) in primary polyp tissues. Under continuous incubation in calcein-containing seawater, initial crystallization and skeletal growth were visualized among the calicoblastic cells in live primary polyp tissues. Additionally, the distribution of calcein-stained SCM and contraction movements of the pockets of SCM were captured at intervals of a few minutes. Our experimental system provided several new insights into coral calcification, particularly as a first step in monitoring the relationship between cellular dynamics and calcification in vivo. Our study suggests that coral calcification initiates at intercellular spaces, a finding that may contribute to the general understanding of coral calcification processes. 相似文献
98.
Aleksandro S. Da Silva Luziane P. Bellé Paula E.R. Bitencourt Herakles A. Garcia Perez Gustavo R. Thomé Marcio M. Costa Camila B. Oliveira Marta M.G. Teixeira Maria B. Moretto Cinthia M. Mazzanti Sonia T.A. Lopes Silvia G. Monteiro 《Experimental parasitology》2011,(1):173-177
The study was undertaken to evaluate changes in the activity of adenosine deaminase (ADA) in brains of rats infected by Trypanosoma evansi. Each rat was intraperitoneally infected with 106 trypomastigotes either suspended in fresh (group A; n = 13) and cryopreserved blood (group B; n = 13). Thirteen animals were used as control (group C). ADA activity was estimated in the cerebellum, cerebral cortex, striatum and hippocampus. No differences (P > 0.05) in ADA activity were observed in the cerebellum between infected and non-infected animals. Significant (P < 0.05) reductions in ADA activity occurred in cerebral cortex in acutely (day 4 post-infection; PI) and chronically (day 20 PI) infected rats. ADA activity was significantly (P < 0.05) decreased in the hippocampus in acutely infected rats, but significantly (P < 0.05) increased in the chronically infected rats. Significant (P < 0.05) reductions in ADA activity occurred in the striatum of chronically infected rats. Parasites could be found in peripheral blood and brain tissue through microscopic examination and PCR assay, respectively, in acutely and chronically infected rats. The reduction of ADA activity in the brain was associated with high levels of parasitemia and anemia in acute infections. Alterations in ADA activity of the brain in T. evansi-infected rats may have implications for pathogenesis of the disease. 相似文献
99.
目的:探讨异氟烷预处理对电磁脉冲辐射所致脑损伤的保护作用。方法:选取成年雄性SD大鼠48只,采用随机数字表法,将其随机分为4组(n=12),分别为:假辐照组(CON组)、电磁辐照组(EMP组)、异氟烷预处理组(IP组)和异氟烷预处理+电磁辐照组(IP+EMP组)。EMP组场强为400 KV/m,脉冲为200次,连续辐照3天;IP组吸入2.0%异氟醚2h;IP+EMP组吸入2.0%异氟醚2 h,24 h后制备EMP损伤模型。于辐照后24 h处死大鼠,每组随机抽取3只大鼠,取脑组织,采用ELISA法检测大鼠海马IL-6和TNF-α的表达变化;尼氏染色法观察大鼠海马区神经元的凋亡;采用Western blot法检测大鼠海马区BDNF蛋白的表达情况;采用免疫荧光法检测大鼠海马区BDNF细胞水平的表达。结果:与CON组比较,EMP组、IP组、IP+EMP组的IL-6和TNF-α的表达增高,尼氏小体减少,BDNF蛋白及细胞水平的表达均下调(P0.05);与EMP组比较,IP组和IP+EMP组IL-6和TNF-α的表达降低,尼氏小体增多,BDNF蛋白及细胞水平的表达上调(P0.05)。结论:异氟烷预处理可减轻电磁脉冲辐射所致脑损伤,其机制可能与减轻大鼠炎症反应有关。 相似文献
100.