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291.
Translational control is a common regulatory mechanism for the expression of iron-related proteins. For example, three enzymes involved in erythrocyte development are regulated by three different control mechanisms: globin synthesis is modulated by heme-regulated translational inhibitor, erythroid 5-aminolevulinate synthase translation is inhibited by binding of the iron regulatory protein to the iron response element in the 5'-untranslated region (UTR); and 15-lipoxygenase is regulated by specific proteins binding to the 3'-UTR. Ceruloplasmin (Cp) is a multi-functional, copper protein made primarily by the liver and by activated macrophages. Cp has important roles in iron homeostasis and in inflammation. Its role in iron metabolism was originally proposed because of its ferroxidase activity and because of its ability to stimulate iron loading into apo-transferrin and iron efflux from liver. We have shown that Cp mRNA is induced by interferon (IFN)-gamma in U937 monocytic cells, but synthesis of Cp protein is halted by translational silencing. The silencing mechanism requires binding of a cytosolic inhibitor complex, IFN-Gamma-Activated Inhibitor of Translation (GAIT), to a specific GAIT element in the Cp 3'-UTR. Here, we describe our studies that define and characterize the GAIT element and elucidate the specific trans-acting proteins that bind the GAIT element. Our experiments describe a new mechanism of translational control of an iron-related protein and may shed light on the role that macrophage-derived Cp plays at the intersection of iron homeostasis and inflammation. 相似文献
292.
Cajanus indicus L, a herb, is popularly known for its hepatoprotective activity. Aqueous extract of the leaves of this plant contains hepatoprotective and hepatostimulatory molecule(s). Present study was aimed to isolate, purify and characterize the active principle(s) responsible for that activity. A hepatoprotective protein molecule has been purified to homogeneity (approximately 300 fold). Homogeneous preparation of the protein was achieved by homogenization, (NH4)2SO4 precipitation, ion-exchange chromatography, gel filtration and high performance liquid chromatography. The protein purified is composed of a single polypeptide chain having an apparent molecular mass of 43 kD as determined by SDS-PAGE and gel filtration through sephadex G-75 column. The isoelectric point of the protein determined was 4.8. Loss of biological activity after heat and protease treatment confirmed that the active molecule is a protein. Peptide fragments of the protein generated by trypsin cleavage were subjected to MALDI-TOF as well as LC-MS analyses and among the various fragments, four were very prominent and used for the determination of the amino acid sequence of the hepatoprotective protein. While one of the peptide fragment revealed strong sequence homology with plastocyanin, another fragment showed some similarity with a tomato protein present in the NCBI non-redundant database. The third peptide, on the other hand, is unique as it did not show any sequence homology with any known protein in the database. The protein showed maximum hepatoprotective activity when administered at a dose of 2 mg/kg body weight for five days after CCl4 administration. Histopathological studies also supported the hepatoprotective nature of the protein. Along with its curative property, the protein also possesses preventive role against a number of toxin induced hepatic damages.Kasturi Sarkar and Ayantika Ghosh contributed equally in the study 相似文献
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294.
Juthika Kundu Rupa Mazumder Ranjana Srivastava & Brahm S. Srivastava 《FEMS immunology and medical microbiology》2009,56(2):179-184
Intranasal immunization, a noninvasive method of vaccination, has been found to be effective in inducing systemic and mucosal immune responses. The present study was aimed at investigating the efficacy of intranasal immunization in inducing mucosal immunity in experimental cholera by subunit recombinant protein vaccines from Vibrio cholerae O1. The structural genes encoding toxin-coregulated pilus A (TcpA) and B subunit of cholera toxin (CtxB) from V. cholerae O1 were cloned and expressed in Escherichia coli . Rabbits were immunized intranasally with purified TcpA and CtxB alone or a mixture of TcpA and CtxB. Immunization with TcpA and CtxB alone conferred, respectively, 41.1% and 70.5% protection against V. cholerae challenge, whereas immunization with a mixture of both antigens conferred complete (100%) protection, as assayed in the rabbit ileal loop model. Serum titers of immunoglobulin G (IgG) antibodies to TcpA and CtxB, and anti-TcpA- and anti-CtxB-specific sIgA in intestinal lavage of vaccinated animals were found to be significantly elevated compared with unimmunized controls. Vibriocidal antibodies were detected at remarkable levels in rabbits receiving TcpA antigen and their titers correlated with protection. Thus, mucosal codelivery of pertinent cholera toxoids provides enhanced protection against experimental cholera. 相似文献
295.
R Mazumder 《European journal of biochemistry》1975,58(2):549-554
1. A ribosome-independent GTPase activity has been isolated from the high-speed supernatant fraction of Artemia salina embryos, and some of its properties have been studied. This activity is inhibited by fusidic acid, an antibiotic generally thought to inhibit only EF-2 in eukaryotes. However, several lines of evidence indicate that the GTPase activity, described here, is distinct from EF-2. The results suggest, therefore, that the inhibitory effect of fusidic acid in eukaryotic systems is not restricted to EF-2 (and ribosome)-dependent functions only. 2. The results of other experiments have revealed that, despite its ability to inhibit the GTPase activity mentioned above, fusidic acid is not a non-specific inhibitor of all ribosome-independent GTPase and ATPase activities present in eukaryotic cells. 相似文献
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298.
The association between glutamate decarboxylase (GAD) and its cofactor, pyridoxal-5′-phos-phate (pyridoxal-P), was studied using 20,0000 supernatant of rat brain. In this preparation GAD required added pyridoxal-P to maintain a linear reaction rate beyond 5 min of incubation. Following exhaustive dialysis the enzyme was more than 83% saturated with cofactor indicating that the cofactor was tightly bound to the enzyme. When incubations were performed in the presence of glutamate and without added pyridoxal-P there was a progressive inactivation of the enzyme which was dependent on the glutamate concentration. This lost activity was almost completely recovered by addition of pyridoxal-P to the dialyzed glutamate-inactivated enzyme. The results suggest that glutamate inactivates GAD by promoting the dissociation of pyridoxal-P from the enzyme thereby producing inactive apoen-zyme which can be reactivated by combining with available pyridoxal-P. This interpretation is supported by the finding that progress curves for the reaction were accurately described over a 30 min incubation period and 10-fold glutamate concentration range by an integrated rate equation which takes the glutamate-promoted dissociation of cofactor into account. The progressive inactivation could not be attributed to denaturation of the enzyme, impurities in the substrate, effects of pH, depletion of substrate, protein concentration, sulfhydryl reagents or product inhibition. The results presented here also show that certain precautions must be adopted to accurately measure GAD activity in the absence of added pyridoxal-P as has been widely done in studies of drug action. Specifically, measurements must be made at short times of incubation and low concentrations of glutamate to minimize the glutamate-promoted inactivation of the enzyme. 相似文献
299.
The naturally occurring free and glycosylated flavans are reviewed. The distribution, methods of isolation and structure elucidation of flavans, by comprehensive spectral analyses, are presented. Biological properties of flavans are appraised. 相似文献
300.
U K Mazumder M Gupta A Mukherjee D K Mukhopadhyay P Dey 《Indian journal of experimental biology》1999,37(7):667-670
Through energy minimization using molecular mechanics force field four ruthenium cordinate complexes have been synthesized. Compound I to IV showed antineoplastic activity with varying degree on EAC bearing mice. Mode of action may be through inhibition of antioxidant property of tumor cell as evident from lipid peroxidase activity. Among the complexes Bis pyridine tetrachloro ruthenium exhibits highest order of activity with respect to increase mean survival time, inhibition of tumour volume, total blood count, hemoglobin and lipid peroxidase activity. 相似文献