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991.
Rognoni Franco Gianesello Daniela Maddalena Fabrizio Giuntini Ilaria Herbst Detlev 《Cytotechnology》1999,29(1):11-25
Anti PSA monoclonal antibodies for diagnostic use were produced in an in vitro system. After purification using Protein G affinity chromatography a percentage of about 10% of antibody aggregates remained. The use of monoclonal antibodies containing aggregates as a capture antibody in a diagnostic kit reduces the performance of the test making it often unacceptable. The aggregates could be eliminated using gel filtration chromatography but, in that way, the final recovery of the whole production process was only about 50%. Aggregation is favoured when the working pH is near to the isoelectric point of the antibody. We varied the culture medium composition, modifying pH and osmolarity. We tested different values of pH and osmolarity: 7.1, 7.5, 8.0, 8.5 for pH, and 300, 340, 367, 395 mOsm/kg H2O for osmolarity. By modification of the cell culture medium we obtained a significant decrease of monoclonal antibody aggregates in the production cycle. In this way we achieved higher recovery rate and could avoid gel filtration polishing step. The experiments were performed in two stages: first in culture flasks changing one parameter in each experiment, and then in spinner bottle using the best conditions obtained in the first stage. During scale up we used the modifications achieved from the experiment showed in this paper in our production by hollow fibre bioreactor with positive results. 相似文献
992.
Nicéia Spanholi Calgaroto Gustavo Roberto Thomé Pauline da Costa Jucimara Baldissareli Fátima Abdala Hussein Roberta Schmatz Maribel A. Rubin Cristiane Signor Daniela Aymone Ribeiro Fabiano Barbosa Carvalho Lizielle Souza de Oliveira Luciane Belmonte Pereira Vera Maria Morsch Maria Rosa Chitolina Schetinger 《Cell biochemistry and function》2014,32(6):502-510
Diabetes is associated with long‐term complications in the brain and reduced cognitive ability. Vitamin D3 (VD3) appears to be involved in the amelioration of hyperglycaemia in streptozotocin (STZ)‐induced diabetic rats. Our aim was to analyse the potential of VD3 in avoiding brain damage through evaluation of acetylcholinesterase (AChE), Na+K+‐adenosine triphosphatase (ATPase) and delta aminolevulinate dehydratase (δ‐ALA‐D) activities and thiobarbituric acid reactive substance (TBARS) levels from cerebral cortex, as well as memory in STZ‐induced diabetic rats. Animals were divided into eight groups (n = 5): control/saline, control/metformin (Metf), control/VD3, control/Metf + VD3, diabetic/saline, diabetic/Metf, diabetic/VD3 and diabetic/Metf + VD3. Thirty days after treatment, animals were submitted to contextual fear‐conditioning and open‐field behavioural tests, after which they were sacrificed and the cerebral cortex was dissected. Our results demonstrate a significant memory deficit, an increase in AChE activity and TBARS levels and a decrease in δ‐ALA‐D and Na+K+‐ATPase activities in diabetic rats when compared with the controls. Treatment of diabetic rats with Metf and VD3 prevented the increase in AChE activity when compared with the diabetic/saline group. In treated diabetic rats, the decrease in Na+K+‐ATPase was reverted when compared with non‐treated rats, but the increase in δ‐ALA‐D activity was not. VD3 prevented diabetes‐induced TBARS level and improved memory. Our results show that VD3 can avoid cognitive deficit through prevention of changes in important enzymes such as Na+K+‐ATPase and AChE in cerebral cortex in type 1 diabetic rats. Copyright © 2014 John Wiley & Sons, Ltd. 相似文献
993.
994.
Shereen Hasan Ghamartaj Hosseini Marc Princivalle Ji-Cui Dong Daniela Birsan Cristina Cagide Ariane I de Agostini 《Biology of reproduction》2002,66(1):144-158
During the reproductive cycle, ovarian follicles undergo major tissue-remodeling involving vascular changes and proteolysis. Anticoagulant heparan sulfate proteoglycans (aHSPGs) are expressed by granulosa cells during the development of the ovarian follicle. The function of aHSPGs in the ovary is unknown, but they might be involved in proteolysis control through binding and activation of serine protease inhibitors. To identify functional interactions between aHSPGs and heparin-binding protease inhibitors in the follicle, we have coordinately localized aHSPGs, antithrombin III, protease nexin-1, and plasminogen activator inhibitor-1 in the rat ovary during natural and gonadotropin-stimulated cycles. Anticoagulant HSPGs were visualized by autoradiography of cryosections incubated with 125I-antithrombin III, and protease inhibitors were assessed by immunohistochemistry and Northern blot hybridization. Anticoagulant HSPGs were expressed in follicles before ovulation, were transiently decreased in postovulatory follicles, and were abundant in the corpus luteum, mainly on capillaries. Anticoagulant HSPGs were colocalized with protease nexin-1 in follicles from the early antral stage until ovulation, with antithrombin III in the preovulatory stage and after ovulation, and with plasminogen activator inhibitor-1 in the corpus luteum. These data demonstrate that aHSPGs are critically expressed in the ovary to interact sequentially with protease nexin-1, antithrombin III, and plasminogen activator inhibitor-1 during the cycle. The specificity of these inhibitors is shifted toward thrombin inhibition in the presence of heparin, suggesting that aHSPGs direct their action to control fibrin deposition in the follicle. The occupation of aHSPGs antithrombin-binding sites by mutant R393C antithrombin III, injected in the ovarian bursa, decreased ovulation efficiency, further supporting the involvement of aHSPGs in the ovulation process. 相似文献
995.
An hybrid experiment, composed of 1H-NMR total correlation spectroscopy (TOCSY) and rotating frame nuclear Overhauser enhancement spectroscopy (ROESY) steps, makes it possible to determine both the partial (disaccharide) sequences, and the sequential order, of whole linear and branched oligosaccharide chains. 相似文献
996.
Laura Andolfi Eugenia Bourkoula Elisa Migliorini Anita Palma Anja Pucer Miran Skrap Giacinto Scoles Antonio Paolo Beltrami Daniela Cesselli Marco Lazzarino 《PloS one》2014,9(11)
Active cell migration and invasion is a peculiar feature of glioma that makes this tumor able to rapidly infiltrate into the surrounding brain tissue. In our recent work, we identified a novel class of glioma-associated-stem cells (defined as GASC for high-grade glioma -HG- and Gasc for low-grade glioma -LG-) that, although not tumorigenic, act supporting the biological aggressiveness of glioma-initiating stem cells (defined as GSC for HG and Gsc for LG) favoring also their motility. Migrating cancer cells undergo considerable molecular and cellular changes by remodeling their cytoskeleton and cell interactions with surrounding environment. To get a better understanding about the role of the glioma-associated-stem cells in tumor progression, cell deformability and interactions between glioma-initiating stem cells and glioma-associated-stem cells were investigated. Adhesion of HG/LG-cancer cells on HG/LG-glioma-associated stem cells was studied by time-lapse microscopy, while cell deformability and cell-cell adhesion strengths were quantified by indentation measurements by atomic force microscopy and single cell force spectroscopy. Our results demonstrate that for both HG and LG glioma, cancer-initiating-stem cells are softer than glioma-associated-stem cells, in agreement with their neoplastic features. The adhesion strength of GSC on GASC appears to be significantly lower than that observed for Gsc on Gasc. Whereas, GSC spread and firmly adhere on Gasc with an adhesion strength increased as compared to that obtained on GASC. These findings highlight that the grade of glioma-associated-stem cells plays an important role in modulating cancer cell adhesion, which could affect glioma cell migration, invasion and thus cancer aggressiveness. Moreover this work provides evidence about the importance of investigating cell adhesion and elasticity for new developments in disease diagnostics and therapeutics. 相似文献
997.
Giulia Bernardini Gemma Leone Lia Millucci Marco Consumi Daniela Braconi Ottavia Spiga Silvia Galderisi Barbara Marzocchi Cecilia Viti Giovanna Giorgetti Pietro Lupetti Agnese Magnani Annalisa Santucci 《Journal of cellular physiology》2019,234(5):6696-6708
Alkaptonuria (AKU) is a disease caused by a deficient homogentisate 1,2-dioxygenase activity leading to systemic accumulation of homogentisic acid (HGA), that forms a melanin-like polymer that progressively deposits onto connective tissues causing a pigmentation called “ochronosis” and tissue degeneration. The effects of AKU and ochronotic pigment on the biomechanical properties of articular cartilage need further investigation. To this aim, AKU cartilage was studied using thermal (thermogravimetry and differential scanning calorimetry) and rheological analysis. We found that AKU cartilage had a doubled mesopore radius compared to healthy cartilage. Since the mesoporous structure is the main responsible for maintaining a correct hydrostatic pressure and tissue homoeostasis, drastic changes of thermal and rheological parameters were found in AKU. In particular, AKU tissue lost its capability to enhance chondrocytes metabolism (decreased heat capacity) and hence the production of proteoglycans. A drastic increase in stiffness and decrease in dissipative and lubricant role ensued in AKU cartilage. Multiphoton and scanning electron microscopies revealed destruction of cell–matrix microstructure and disruption of the superficial layer. Such observations on AKU specimens were confirmed in HGA-treated healthy cartilage, indicating that HGA is the toxic responsible of morphological and mechanical alterations of cartilage in AKU. 相似文献
998.
999.
Maurício Luiz Vilela Daniela de Pita-Pereira Carina Graser Azevedo Rodrigo Espíndola Godoy Constan?a Britto Elizabeth Ferreira Rangel 《Memórias do Instituto Oswaldo Cruz》2013,108(5):578-585
Phlebotomine sandflies were captured in rural settlement and periurban
areas of the municipality of Guaraí in the state of Tocantins (TO), an endemic
area of American cutaneous leishmaniasis (ACL). Forty-three phlebotomine species
were identified, nine of which have already been recognised as ACL vectors.
Eleven species were recorded for the first time in TO. Nyssomyia
whitmani was the most abundant species, followed by
Evandromyia bourrouli, Nyssomyia antunesi
and Psychodopygus complexus. The Shannon-Wiener diversity index
and the evenness index were higher in the rural settlement area than in the
periurban area. The evaluation of different ecotopes within the rural area
showed the highest frequencies of Ev. bourrouli and Ny.
antunesi in chicken coops, whereas Ny. whitmani
predominated in this ecotope in the periurban area. In the rural settlement
area, Ev. bourrouli was the most frequently captured species in
automatic light traps and Ps. complexus was the most prevalent
in Shannon trap captures. The rural settlement environment exhibited greater
phlebotomine biodiversity than the periurban area. Ps.
complexus and Psychodopygus ayrozai naturally
infected with Leishmania (Viannia) braziliensis were
identified. The data identified Ny. whitmani as a potential ACL
vector in the periurban area, whereas Ps. complexus was more
prevalent in the rural environment associated with settlements. 相似文献
1000.
Markus Auer Clemens Gruber Marzia Bellei Katharina F. Pirker Marcel Zamocky Daniela Kroiss Stefan A. Teufer Stefan Hofbauer Monika Soudi Gianantonio Battistuzzi Paul G. Furtmüller Christian Obinger 《The Journal of biological chemistry》2013,288(38):27181-27199
Reconstructing the phylogenetic relationships of the main evolutionary lines of the mammalian
peroxidases lactoperoxidase and myeloperoxidase revealed the presence of novel bacterial heme
peroxidase subfamilies. Here, for the first time, an ancestral bacterial heme peroxidase is shown to
possess a very high bromide oxidation activity (besides conventional peroxidase activity). The
recombinant protein allowed monitoring of the autocatalytic peroxide-driven formation of covalent
heme to protein bonds. Thereby, the high spin ferric rhombic heme spectrum became similar to
lactoperoxidase, the standard reduction potential of the Fe(III)/Fe(II) couple shifted to more
positive values (−145 ± 10 mV at pH 7), and the conformational and thermal stability
of the protein increased significantly. We discuss structure-function relationships of this new
peroxidase in relation to its mammalian counterparts and ask for its putative physiological
role. 相似文献