全文获取类型
收费全文 | 3268篇 |
免费 | 305篇 |
国内免费 | 9篇 |
出版年
2022年 | 28篇 |
2021年 | 57篇 |
2020年 | 25篇 |
2019年 | 34篇 |
2018年 | 46篇 |
2017年 | 22篇 |
2016年 | 53篇 |
2015年 | 112篇 |
2014年 | 115篇 |
2013年 | 181篇 |
2012年 | 209篇 |
2011年 | 205篇 |
2010年 | 118篇 |
2009年 | 106篇 |
2008年 | 140篇 |
2007年 | 151篇 |
2006年 | 146篇 |
2005年 | 135篇 |
2004年 | 145篇 |
2003年 | 148篇 |
2002年 | 134篇 |
2001年 | 59篇 |
2000年 | 49篇 |
1999年 | 62篇 |
1998年 | 47篇 |
1997年 | 23篇 |
1996年 | 26篇 |
1995年 | 27篇 |
1994年 | 22篇 |
1993年 | 23篇 |
1992年 | 56篇 |
1991年 | 50篇 |
1990年 | 40篇 |
1989年 | 35篇 |
1988年 | 60篇 |
1987年 | 40篇 |
1986年 | 45篇 |
1985年 | 41篇 |
1984年 | 30篇 |
1983年 | 27篇 |
1982年 | 26篇 |
1981年 | 39篇 |
1980年 | 34篇 |
1979年 | 30篇 |
1978年 | 37篇 |
1977年 | 30篇 |
1976年 | 35篇 |
1975年 | 32篇 |
1974年 | 30篇 |
1972年 | 25篇 |
排序方式: 共有3582条查询结果,搜索用时 187 毫秒
981.
982.
Summary Lysosomal diseases result from deficiency of one of the many enzymes involved in the normal, step-wise breakdown of macromolecules. Studies in vitro have shown that cells from enzyme-deficient patients can be corrected by an exogenous supply of the missing enzyme. This occurs by receptor-mediated endocytosis of normal enzyme added to tissue culture medium and also by direct transfer from normal leukocytes during cell-to-cell contact. Immunohistochemical analysis has revealed that these processes have similar pathways of intracellular transport of the acquired enzymes, which ultimately reach mature lysosomes in the recipient cells. Moreover, recent studies suggest that both mechanisms are important in the therapy of lysosomal storage diseases by bone marrow transplantation. Advances in gene technology are likely to improve the successful treatment of these disorders, by facilitating the large scale production of clinically effective proteins and also by enabling the stable and safe introduction of normal lysosomal genes into cells of affected patients. 相似文献
983.
Young Hoon Oh Matthew L. Becker Kerri M. Mendola Leila H. Choe Lie Min Kelvin H. Lee Yinges Yigzaw Alexander Seay Jerome Bill Xuanwen Li David J. Roush Steven M. Cramer Stefano Menegatti Abraham M. Lenhoff 《Biotechnology and bioengineering》2023,120(4):1068-1080
In the production of biopharmaceuticals such as monoclonal antibodies (mAbs) and vaccines, the residual amounts of host-cell proteins (HCPs) are among the critical quality attributes. In addition to overall HCP levels, individual HCPs may elude purification, potentially causing issues in product stability or patient safety. Such HCP persistence has been attributed mainly to biophysical interactions between individual HCPs and the product, resin media, or residual chromatin particles. Based on measurements on process streams from seven mAb processes, we have found that HCPs in aggregates, not necessarily chromatin-derived, may play a significant role in the persistence of many HCPs. Such aggregates may also hinder accurate detection of HCPs using existing proteomics methods. The findings also highlight that certain HCPs may be difficult to remove because of their functional complementarity to the product; specifically, chaperones and other proteins involved in the unfolded protein response (UPR) are disproportionately present in the aggregates. The methods and findings described here expand our understanding of the origins and potential behavior of HCPs in cell-based biopharmaceutical processes and may be instrumental in improving existing techniques for HCP detection and clearance. 相似文献
984.
Effects of polyamine depletion by -difluoromethylornithine (DFMO) were studied on the GABA-agonist mediated enhancement of the morphological development of cultured rat cerebellar granule cells. An increase in the number of neurite extending cells and in the cytoplasmic density of organelles relevant for protein synthesis was observed upon culturing in the presence of 4,5,6,7-tetrahydro-isoxazolo[5,4-c]pyridin-3-ol (THIP) for 4 days. The intracellular concentrations of putrescine, spermadine, and spermine in these cultures were similar to the concentrations of the polyamines observed in cultures grown in a plain culture medium for 1, 2, 3 or 4 days, respectively. Upon culturing in the simultaneous presence of THIP and DFMO, the concentrations of putrescine and spermadine were reduced to less than 20% of the levels in the controls. This depletion was associated with a severely impaired morphological development of the granule cell cultures. Thus, the number of neurite extending cells was reduced to 50% of the number in the control cultures upon culturing in the presence of DFMO alone or in combination with THIP. Moreover, the THIP mediated increase in the cytoplasmic density of rough endoplasmic reticulum, Golgi apparatus and different types of vesicles was prevented by the exposure to DFMO. 相似文献
985.
Cesia Shenberg Miriam Boazi Jacob Cohen Abraham Klein Marcus Kojler Abraham Nyska 《Biological trace element research》1994,40(2):137-149
The effect ofcis-DDP treatment with and without selenite and selenocistamine was studied on kidneys of tumor-bearing mice. The amounts ofcis-DDP, selenite, and selenocistamine injected were chosen so as to be compatible with the treatment of humans. The animals
were sacrificed at 7, 14, and 28 d after treatment. The kidneys were removed and subjected to trace element analysis by a
novel X-ray fluorescence (XRF) method and for pathological assessment. The results show that following treatment withcis-DDP, K, Fe, Cu, and Zn reach a maximum level after 7 d; K, Fe, and Cu levels were significantly reduced by the addition of
selenite. The level of Zn was reduced only in groups treated with selenite whereas that of K and Cu was reduced also in groups
treated with selenocistamine with and withoutcis-DDP. The greatest increase in Pt and Se levels was reached 1 wk after injection withcis-DDP, with and without selenocompounds, and in the case of Pt was partly reduced by addition of selenite. Se returned to control
values 2 wk after injection, although Pt was still high in all groups 2 and 4 wk after injection. The results corroborate
the findings of our previous studies. The effect of selenocistamine incis-DDP treated mice was partly insufficient. The pathological examination of the kidneys did not show any differences in the
effect of various additives during the study. 相似文献
986.
The specific activity (units activity/mg cytosolic protein) of malic enzyme was found to be three-fold higher in the livers of mice fed a semipurified diet containing 50% () glucose and 15% () saturated and monounsaturated but no polyunsaturated fat (hydrogenated cottonseed oil) over an 11-day period than in the livers of mice fed a standard laboratory mouse chow (Purina) diet. In contrast, when other lab chow-fed mice were fed an isocaloric diet containing 15% () polyunsaturated fat (corn oil), no change in the specific activity of malic enzyme occurred over a similar period of time. Rocket immunoelectrophoresis performed on cytosols from both dietary groups demonstrated that the livers of mice consuming the hydrogenated cottonseed oil diet contained approximately three times more malic enzyme protein than did the livers from the corn oil-fed animals. In mice pulse-labeled with l-[4,5-3H]leucine, the rate of hepatic malic enzyme synthesis (relative to that for total protein) was approximately twofold greater in the hydrogenated cottonseed oil-fed mice than in their corn oil-fed counterparts whereas the rate of hepatic malic enzyme degradation was similar for both groups. Immunotitration of liver malic enzyme from hydrogenated cottonseed oil-fed and corn oil-fed mice revealed identical equivalence points, demonstrating that the catalytic efficiency of mouse liver malic enzyme had not been affected by the type of dietary fat administered. When total liver RNA, isolated from the hydrogenated cottonseed oil- and the corn oil-fed animals, was translated in cell-free translation systems (wheat germ extract and reticulocyte lysate) we found that both dietary treatments had resulted in an increase in the activity of malic enzyme messenger RNA. Furthermore, there were no significant differences between the two dietary groups in this regard. These results suggest that hepatic malic enzyme specific activity in high-carbohydrate polyunsaturated fat-fed mice is regulated principally by dietary-induced changes in the rate of enzyme synthesis and not by the activity of messenger RNA coding for the enzyme. 相似文献
987.
Transport and metabolism of bacilysin and other peptides by suspensions of Staphylococcus aureus 总被引:2,自引:0,他引:2
L-Alanyl-L-tyrosine and glycyl-L-phenylalanine labelled with 14C competed with each other and with the dipeptide antibiotic bacilysin for transport into Staphylococcus aureus NCTC 6571 in a medium which did not support growth. They also competed with other dipeptides and several tripeptides. The fast initial transport ofthe two labelled peptides appeared to show Michaelis-Menten kinetics. Neither was transported into a bacilysin-resistant mutant of S. aureus NCTC 6571, although tyrosine was taken up by the mutant as readily as it was by the parent strain. Uptake of alanyltyrosine or glycylphenylalanine was followed by rapid hydrolysis of the peptide and the excretion of tyrosine or phenylalanine. Glycine liberated from glycylphenylalanine was partly degraded and partly incorporated into the bacterial wall. The behaviour of these dipeptides paralleled the inactivation of bacilysin by suspensions of S. aureus and the appearance of its C-terminal amino acid, anticapsin, in the extracellular fluid. 相似文献
988.
Using synchronous populations obtained by selectively detaching mitotic cells from cultures grown in monolayer, we demonstrate here that Chinese hamster ovary (CHO) cells exhibit a differential sensitivity to mutation induction by UV as a function of position in the cell cycle. When mutation induction to 6-thioguanine (TG) resistance is monitored, several maxima and minima are displayed during cell-cycle traverse, with a major maximum occurring in early S phase. Although cells in S phase are more sensitive to UV-mediated cell lethality than those in G1 or G2/M phases, there is not a strict correlation with induced mutation frequency. Fluence-response curves obtained at several times during the cell cycle yield Dq values approximating 6 J/m2. The primary survival characteristic which varies with cell cycle position is D0, ranging from 2.5 J/m2 at 6 h after mitotic selection to 5.5 J/m2 at 11 h afterward. Based on studies with asynchronous, logarithmically growing populations, as well as those mitotically selected to be synchronous, the optimum phenotypic expression time for induced TG resistance is 7–9 days and is essentially independent of both UV fluence and position in the cell cycle. All isolated mutants have altered hypozanthine—guanine phosphoribosyl transferase (HGPRT) activity, and no difference in the residual level of activity was detected among isolated clones receiving UV radiation during G1, S, or late S/G2 phases of the cell cycle. Changes in cellular morphology during cell-cycle traverse do not contribute to the differential susceptibility to UV-induced mutagenesis. 相似文献
989.
The present paper deals with the histopathological studies of galls of Ustilago hordei (Pers.) on Hordeum vulgare L. The gall formation in the present case is the result of both hyperplastic and hypertrophic activities of the infected host cells. Also the gall formation by U. hordei on. H. vulgare is being reported for the first time through present. communication. 相似文献
990.
Summary Electron microscopic analysis of primary cultures derived from larvalXenopus liver has shown that these cells, although they form only two-dimensional aggregates, retain and presumably also develop structural characteristics typical of liver parenchyma cells, such as bile canaliculi with microvilli and epithelial junctional complexes. As judged from structural criteria, primary cultures contain 80–90% hepatocytes. In contrast to the intact tissue, primary cultures showed excessive development of microfilaments, however.Incorporation of labeled amino acids has revealed further that the capacity for protein synthesis is maintained in culture and that synthesis of liverspecific protein albumin is maintained in vitro, even in liver cultures derived from thyrostatic tadpoles. This latter result suggests that initiation of albumin synthesis in the larval liver is probably not dependent upon thyroid hormones but rather reflects the protodifferentiated state of this tissue. 相似文献