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931.
Duarte Toubarro Miguel Lucena-Robles Gisela Nascimento Gonalo Costa Rafael Montiel Ana V. Coelho Nelson Simes 《International journal for parasitology》2009,39(12):1319-1330
Steinernema carpocapsae is an insect parasitic nematode able to parasitise and kill the host within 48 h. Secreted products (ESP) of the parasitic stage of a virulent strain contain higher amounts of proteolytic activity than a low virulence strain, suggesting proteases are involved in virulence. From the ESP we purified a protein (Sc-SP-3) with a Mr of 30 kDa and a pI of 7 that cleaved the synthetic substrate N-succinyl-Ala-Ala-Pro-Phe-pNA and was inhibited by phenylmethanesulfonyl fluoride, benzamidine and chymostatin, thus indicating that it belongs to the chymotrypsin-like serine protease family. Sc-SP-3 has a Vmax of 0.3 mM min−1 ml−1 and Km of 6.6 × 10−4 M, with maximum activity at pH 8 and 40 °C. The full-length cDNA was obtained using degenerate oligonucleotides for serine proteases. This open reading frame encodes a preproprotein containing a putative signal peptide composed of 16 amino acid residues, a prodomain of 40 residues and a mature protease domain of 261 residues, including the catalytic triad His/Asp/Ser characteristic of trypsin-like serine proteases. The N-terminal sequence and the peptide masses fingerprint obtained by MALDI-TOF–MS for the purified protein matched the cDNA. Gene expression analysis by quantitative real-time-PCR showed that this gene is expressed only during the parasitic stage and that pre-invasive nematodes inside the mid-gut expressed higher amounts of Sc-SP-3 than those that already enter the haemocoel. Sc-SP-3 caused histolysis in the insect mid-gut. In vitro assays demonstrated that Sc-SP-3 digested extracellular proteins and induced apoptosis in Sf9 insect cells, thus suggesting Sc-SP-3 is a multifunctional chymotrypsin-like protease involved in pathogenesis. 相似文献
932.
933.
Diverse endophytic bacteria isolated from a leguminous tree Conzattia multiflora grown in Mexico 总被引:1,自引:0,他引:1
Wang ET Tan ZY Guo XW Rodríguez-Duran R Boll G Martínez-Romero E 《Archives of microbiology》2006,186(4):251-259
Conzattia multiflora is a leguminous tree present only in Mexico and Guatemala. There is no record about its symbiotic or pathogenic microbes. In this study, we found that numerous bacteria with 104–106 individuals per gram of fresh epidermis were distributed in the tissue of this plant. All the bacteria isolated from the Conzattia epidermis were Gram-negative, facultative anaerobic rods and formed yellow or colorless colonies. They were identified as endophytes by inoculation tests. Some of the bacteria could significantly promote the growth of Conzattia seedlings. Nine different groups were defined by PCR-based RFLP, which were classified as Pantoea, Erwinia, Salmonella, Enterobacter, Citrobacter and Klebsiella by the phylogenetic analysis of 16S rRNA genes. The existence of plant-borne lineages of Salmonella indicates that the unexplored plants may harbor some unknown microbes. 相似文献
934.
Harald Prüss Gisela Grosse Irene Brunk Rüdiger W. Veh Gudrun Ahnert-Hilger 《Histochemistry and cell biology》2010,133(3):301-312
The development of the hippocampal network requires neuronal activity, which is shaped by the differential expression and
sorting of a variety of potassium channels. Parallel to their maturation, hippocampal neurons undergo a distinct development
of their ion channel profile. The age-dependent dimension of ion channel occurrence is of utmost importance as it is interdependently
linked to network formation. However, data regarding the exact temporal expression of potassium channels during postnatal
hippocampal development are scarce. We therefore studied the expression of several voltage-gated potassium channel proteins
during hippocampal development in vivo and in primary cultures, focusing on channels that were sorted to the axonal compartment.
The Kv1.1, Kv1.2, Kv1.4, and Kv3.4 proteins showed a considerable temporal variation of axonal localization among neuronal
subpopulations. It is possible, therefore, that hippocampal neurons possess cell type-specific mechanisms for channel compartmentalization.
Thus, age-dependent axonal sorting of the potassium channel proteins offers a new approach to functionally distinguish classes
of hippocampal neurons and may extend our understanding of hippocampal circuitry and memory processing. 相似文献
935.
María Laura Lavaggi Mauricio Cabrera María de los Ángeles Aravena Claudio Olea-Azar Adela López de Ceráin Antonio Monge Gisela Pachón Marta Cascante Ana María Bruno Lía I. Pietrasanta Mercedes González Hugo Cerecetto 《Bioorganic & medicinal chemistry》2010,18(12):4433-4440
Phenazine 5,10-dioxides are prodrugs for antitumor therapy that undergo hypoxic-selective bioreduction to form cytotoxic species. Here we investigate the expanded system benzo[a]phenazine 7,12-dioxides as selective hypoxic cytotoxin-scaffold. The clonogenic survival of V79 cells on aerobic and anaerobic conditions, conduct us to study antiproliferative activity on Caco-2 tumoral cells in normoxia. Electrochemical, DNA-interaction and DNA-damage studies were performed to establish the mode of action. The results demonstrated the potential biological properties of the studied scaffold being derivatives 6–10 structural hits for further chemical-modifications to become into therapeutics for solid tumors. Compounds 6 and 8 with cytotoxicity against V79 cells in both conditions (aerobia and anaerobia) were also cytotoxic against Caco-2 tumoral cells in aerobiosis. 相似文献
936.
Ca2+ channels that underlie mitochondrial Ca2+ transport first reported decades ago have now just recently been precisely characterized electrophysiologically. Numerous data indicate that mitochondrial Ca2+ uptake via these channels regulates multiple intracellular processes by shaping cytosolic and mitochondrial Ca2+ transients, as well as altering the cellular metabolic and redox state. On the other hand, mitochondrial Ca2+ overload also initiates a cascade of events that leads to cell death. Thus, characterization of mitochondrial Ca2+ channels is central to a comprehensive understanding of cell signaling. Here, we discuss recent progresses in the biophysical and electrophysiological characterization of several distinct mitochondrial Ca2+ channels. 相似文献
937.
Polyunsaturated Fatty acids (PUFAs) seem to be helpful in the therapy of depression. Zinc (Zn) may be one co-factor contributing
to their antidepressive effect. Zn acts lipid protective and is a constituent of fatty acid metabolism enzymes. In animals,
an antidepressive effect of Zn was already demonstrated. Therefore, if and how Zn and PUFAs correlate in depressive patients
or in individuals from the general population was investigated. Blood samples were collected from 88 depressive in-patients
and 88 volunteers from the general population matched for age-group and gender (each 32 men and 56 women, 21–70 years) for
measurement of Zn (colorimetric) and of 12 fatty acids (FAs) (by capillary gas-chromatography). Severity of depression in
patients was assessed by Beck Depression Inventory (BDI) and Hamilton Depression Rating Scale (HDRS). Zn concentration was
independent of age, gender and body-mass-index and significantly correlated with the severity of depression measured by BDI
(r = 0.26; P = 0.034) in depressive patients,. HDRS was inversely correlated with gammalinolenic acid concentration (r = −0.24; P = 0.029). Median serum Zn concentration in depressive patients did not differ from control individuals. Zn was correlated
with myristic acid concentration (r = 0.22; P < 0.05) in controls from the general population; and a negative correlation between Zn and dihomogammalinolenic acid concentration
(r = −0.26; P < 0.05) was found in depressive inpatients. FA composition in serum significantly differed between depressive and healthy
persons: Depressive patients had higher stearic and arachidonic acid (AA) concentration. Relative to AA, their eicosapentaenoic
and docosapentaenoic acid concentration were diminished compared to the general populations group. These results do not confirm
the hypothesis of a general lack of Zn in depressive disorders, but Zn concentrations differed dependent on comorbid disorders
and severity of depression. In depressive patients and control persons Zn concentration is associated with different FAs indicating
diverging metabolic pathways. 相似文献
938.
The role of free radicals and active states of oxygen in human cancer is as yet unresolved. Various lines of evidence provide strong but inferential evidence that free radical reactions can be of crucial importance in certain carcinogenic mechanisms. A central point in considering free radical reactions in carcinogenesis is that human cancer is really a group of highly diverse diseases for which the initial causation and the progression to clinical disease occur through a wide variety of mechanisms. Furthermore, for many human cancers it appears that there are alternate pathways capable of tumor initiation and tumor progression. While for certain of these pathways free radical reactions appear necessary, it is unlikely that there are human cancers for which free radicals, or any other mechanism, are sufficient for the entire processbeginning with the genetic alteration leading to a somatic mutation and eventually resulting in clinically overt disease. It is crucial that we view free radical reactions as aong a panoply of mechanisms leading to human cancer, and consider research about the role of free radicals in cancer as opportunities to prevent the initiation or progression of human cancer. 相似文献
939.
Identification of Mycelium-Associated Cellulase from Streptomyces reticuli 总被引:2,自引:3,他引:2
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Gisela Wachinger Karin Bronnenmeier Walter L. Staudenbauer Hildgund Schrempf 《Applied microbiology》1989,55(10):2653-2657
Among 180 Streptomyces strains tested, 25 were capable of hydrolyzing microcrystalline cellulose (Avicel) at 30°C. Streptomyces reticuli was selected for further studies because of its ability to grow at between 30 and 50°C on Avicel. Enzymatic activities degrading Avicel, carboxymethyl cellulose, and cellobiose were found both in the culture supernatant and in association with the mycelium and crystalline substrate. The bound enzymes were efficiently solubilized by repeated washes with buffer of low ionic strength (50 mM Tris hydrochloride [pH 7.5]) and further purified by fast protein liquid chromatography. A high-molecular-weight Avicelase of >300 kilodaltons could be separated from carboxymethyl cellulase (CMCase) and β-glucosidase activities (molecular mass, 40 to 50 kilodaltons) by gel filtration on Superose 12. The CMCase fraction was resolved by Mono Q anion-exchange chromatography into two enzymes designated CMCase 1 and CMCase 2. The β-glucosidase activity was found to copurify with CMCase 2. The purified cellulase components showed optimal activity at around pH 7.0 and temperatures of between 45 and 50°C. Avicelase (but not CMCase) activity was stimulated significantly by the addition of CaCl2. 相似文献
940.
Due to the ongoing development of clinical photodynamic therapy (PDT), the search continues for optimized photosensitizers that can overcome some of the side effects associated with this type of treatment modality. The main protagonists being: post-treatment photosensitivity, due to only limited cellular selectivity and post-treatment tumor regrowth, due to the up-regulation of pro-inflammatory agents within the tumor microenvironment. A photosensitizer that could overcome one or both of these drawbacks would be highly attractive to those engaged in clinical PDT. Certain non-steroidal anti-inflammatory drugs (NSAIDs) when used in combination with PDT have shown to increase the cytotoxicity of the treatment modality by targeting the tumor microenvironment. Temoporfin (m-THPC), the gold standard chlorin-based photosensitizer (PS) since its discovery in the 1980’s, has successfully been conjugated to non-steroidal anti-inflammatory compounds, in an attempt to address the issue of post-treatment tumor regrowth. Using a modified Steglich esterification reaction, a library of “iPorphyrins” was successfully synthesized and evaluated for their PDT efficacy. 相似文献