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141.
Yasuko Koshihara Mieko Yamagishi Sei-Itsu Murota 《Biochimica et Biophysica Acta (BBA)/General Subjects》1976,437(1):221-228
The binding activity of [3H]dexamethasone to the specific receptor was studied in the cytoplasmic fraction of a established fibroblast line derived from rat carrageenin granuloma in culture condition. Specific receptor to dexamethasone was demonstrated. Scatchard analysis revealed a single class of binding sites with a dissociation constant for [3H]dexamethasone of 3.64 · 10?8 M and a concentration of binding sites of 0.825 pmol per mg cytosol protein. The number of cytoplasmic binding sites per cell was calculated at 1.15 · 105.Total binding activity to [3H]dexamethasone of the cytoplasmic fraction was enhanced when the cells were cultured in a medium containing salicylic acid at 37°C. The maximum enhancement was seen at the concentration of 10?3 M and in 3 h treatment of salicylic acid. This enhancement by salicylic acid was lost when cycloheximide was added to the culture medium at the same time. If salicylic acid was added to the cell free system, it showed no effect on the binding activity. The other non-steroidal anti-inflammatory drugs; phenylbutazone and indomethacin, also enhanced the total binding activity to [3H]dexamethasone of the cytoplasmic fraction at the concentration of 2 · 10?5 M and 2 · 10?7 M, respectively. 相似文献
142.
Enhancement of 5-lipoxygenase activity in mastocytoma P-815 cells by n-butyrate treatment 总被引:1,自引:0,他引:1
Cloned mastocytoma P-815, 2-E-6 cells were used to investigate regulation of 5-lipoxygenase activity. 2-E-6 cells had high 5-lipoxygenase activity with slight 12-lipoxygenase activity. The 5-lipoxygenase activity was increased over 5-fold by treatment of the cells with 1 mM n-butyrate for 18 h. the most effective dose range being 0.1-5.0 mM. Treatment with n-butyrate for 18 h was more effective than treatment for 40 h. Addition of n-butyrate to an untreated cell homogenate had no stimulatory effect. The enhancement of 5-lipoxygenase activity by n-butyrate was accompanied by new synthesis of protein(s). 12-Lipoxygenase activity was not increased so much as 5-lipoxygenase activity by the treatment. This is the first report of stimulation of 5-lipoxygenase activity in cultured cells. The different responses of the two lipoxygenases to n-butyrate treatment strongly suggest that 5-lipoxygenase is a different enzyme from 12-lipoxygenase. 相似文献
143.
A new method for the isolation of a high yield of collagen from human skeletal muscle is described. This technique employs rapid stirring of the homogenate of skeletal muscle with a magnetic stirrer. Fibrous material entangled during rapid stirring was recovered by passing the homogenate through a sieve and then sequentially extracted with Hasselbach-Schneider solution and 0.6 m KI-0.06 m Na2S2O3. The insoluble residue obtained after these extractions was shown to be highly purified collagen by amino acid analysis, and the recovery of collagen by this method was found to be more than 90% of the total collagen in skeletal muscle. The isolated collagen from 5 years of age was mostly composed of Type I collagen, and small amounts of Type III collagen and an unidentified collagenous protein migrating in a position near Type V collagen (αB chain) were also found on a sodium dodecyl sulfate-polyacrylamide gel. 相似文献
144.
An improved laser microprobe procedure is developed and applied to the measurement of calcium content in microareas of right and left subepicardial muscles. The staining of canine cardiac muscles by Methylene Blue solution (1% w/v) was found to improve sampling efficiency. Elemental content is proportional to T-1/gamma, where T is the transmittance of the characteristic emission line of the element of the photographic plate and gamma is its contrast. In the present system, the calcium content is analyzed using T-3.2. We find that the staining of samples and the determination of T-1/gamma are useful procedures in the application of laser microprobe to the study of elemental content in biologic microareas. 相似文献