全文获取类型
收费全文 | 2144篇 |
免费 | 130篇 |
国内免费 | 1篇 |
出版年
2022年 | 9篇 |
2021年 | 19篇 |
2020年 | 16篇 |
2019年 | 27篇 |
2018年 | 15篇 |
2017年 | 28篇 |
2016年 | 45篇 |
2015年 | 70篇 |
2014年 | 82篇 |
2013年 | 124篇 |
2012年 | 141篇 |
2011年 | 144篇 |
2010年 | 76篇 |
2009年 | 94篇 |
2008年 | 131篇 |
2007年 | 141篇 |
2006年 | 121篇 |
2005年 | 117篇 |
2004年 | 117篇 |
2003年 | 146篇 |
2002年 | 130篇 |
2001年 | 18篇 |
2000年 | 19篇 |
1999年 | 25篇 |
1998年 | 29篇 |
1997年 | 23篇 |
1996年 | 16篇 |
1995年 | 22篇 |
1994年 | 18篇 |
1993年 | 17篇 |
1992年 | 23篇 |
1991年 | 12篇 |
1990年 | 15篇 |
1989年 | 12篇 |
1988年 | 11篇 |
1987年 | 19篇 |
1986年 | 17篇 |
1985年 | 9篇 |
1984年 | 10篇 |
1981年 | 8篇 |
1980年 | 8篇 |
1979年 | 12篇 |
1978年 | 18篇 |
1977年 | 10篇 |
1976年 | 8篇 |
1975年 | 8篇 |
1974年 | 9篇 |
1973年 | 10篇 |
1972年 | 9篇 |
1971年 | 10篇 |
排序方式: 共有2275条查询结果,搜索用时 15 毫秒
71.
Abstract Recent site-directed mutagenesis and thermodynamic studies have shown that the V74I mutant of Escherichia coli ribonuclease HI (RNase HI) is more stable than the wild type protein [Ishikawa et al., Biochemistry 32, 6171 (1993)]. In order to clarify the stabilization mechanism of this mutant, we calculated the free energy change due to the mutation Val 74→Ile in both the native and denatured states by free energy perturbations based on molecular dynamics (MD) simulations. We carried out inclusive MD simulations for the protein in water; i.e., fully solvated, no artificial constraints applied, and all long-range Coulomb interactions included. We found that the free energy of the mutant increased slightly relative to the wild type, in the native state by 1.60 kcal/mol, and in the denatured state by 2.25 kcal/mol. The unfolding free energy increment of the mutant (0.66 ± 0.19 kcal/mol) was in good agreement with the experimental value (0.6 kcal/mol). The hysteresis error in the free energy calculations, i.e., forward and reverse perturbations, was only ±0.19 kcal/mol. These results show that the V74I mutant is stabilized relative to the wild type by the increased free energy of the denatured state and not by a decrease in the free energy of the native state as had been proposed earlier based on the mutant X-ray structure. It was found that the stabilization was caused by a loss of solvation energy in the mutant denatured state and not by improved packing interactions inside the native protein. 相似文献
72.
Hirokazu Saiwai Hiromi Kumamaru Yasuyuki Ohkawa Kensuke Kubota Kazu Kobayakawa Hisakata Yamada Takehiko Yokomizo Yukihide Iwamoto Seiji Okada 《Journal of neurochemistry》2013,125(1):74-88
Acute inflammation is a prominent feature of central nervous system (CNS) insult and is detrimental to the CNS tissue. Although this reaction spontaneously diminishes within a short period of time, the mechanism underlying this inflammatory resolution remains largely unknown. In this study, we demonstrated that an initial infiltration of Ly6C+Ly6G? immature monocyte fraction exhibited the same characteristics as myeloid‐derived suppressor cells (MDSCs), and played a critical role in the resolution of acute inflammation and in the subsequent tissue repair by using mice spinal cord injury (SCI) model. Complete depletion of Ly6C+Ly6G? fraction prior to injury by anti‐Gr‐1 antibody (clone: RB6‐8C5) treatment significantly exacerbated tissue edema, vessel permeability, and hemorrhage, causing impaired neurological outcomes. Functional recovery was barely impaired when infiltration was allowed for the initial 24 h after injury, suggesting that MDSC infiltration at an early phase is critical to improve the neurological outcome. Moreover, intraspinal transplantation of ex vivo‐generated MDSCs at sites of SCI significantly reduced inflammation and promoted tissue regeneration, resulting in better functional recovery. Our findings reveal the crucial role of an Ly6C+Ly6G? fraction as MDSCs in regulating inflammation and tissue repair after SCI, and also suggests an MDSC‐based strategy that can be applied to acute inflammatory diseases. 相似文献
73.
Shunichi Kojima Masato KakuToshitsugu Kawata Hiromi SumiHanaka Shikata Tahsin Raquib AbontiShotoku Kojima Tadashi FujitaMasahide Motokawa Kazuo Tanne 《Cryobiology》2013
Mesenchymal stem cells (MSCs) can be used for the regeneration of various tissues and cryopreservation of MSCs is so important for regenerative medicine. The purpose of this study was to evaluate the influences of cryopreservation on MSCs by use of a programmed freezer with a magnetic field (CAS freezer). MSCs were isolated from bone marrow of rat femora. The cells were frozen by a CAS freezer with 10% dimethyl sulfoxide (Me2SO) and cryopreserved for 7 days at a temperature of −150 °C. Immediately after thawing, the number of survived cells was counted. The cell proliferation also examined after 48 h culture. Next, MSCs were frozen by two different freezers; CAS freezer and a conventional programmed freezer without magnetic field. Then, osteogenic and adipogenic differentiations of cryopreserved cells were examined. As a result, survival and proliferation rates of MSCs were significantly higher in CAS freezer than in the non-magnetic freezer. Alizarin positive reaction, large amount of calcium quantification, and greater alkaline phosphatase activity were shown in both the non-cryopreserved and CAS groups after osteogenic differentiation. Moreover, Oil Red O staining positive reaction and high amount of PPARγ and FABP4 mRNAs were shown in both the non-cryopreserved and CAS groups after adipogenic differentiation. From these findings, it is shown that a CAS freezer can maintain high survival and proliferation rates of MSCs and maintain both adipogenic and osteogenic differentiation abilities. It is thus concluded that CAS freezer is available for cryopreservation of MSCs, which can be applied to various tissue regeneration. 相似文献
74.
Takashi Kitayama Kanako Yasuda Takeharu Kihara Michiho Ito Hiromi Fukumoto Masanori Morimoto 《Applied Entomology and Zoology》2013,48(4):455-459
Piper ribersoides Wall. (Piperaceae), which is called “Khua Sa khan” in the local language, is mainly grown in Laos. This plant is used as a food in Laos, but no report on its metabolites exists. Crushed stems were immersed in methanol. The ethyl acetate fraction showed potential insect antifeedant activity for Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae), and some of the active compounds were piperine analogs. Piperine and its geometrical isomer independently showed potent antifeedant activities, and piperine was presumably the main active compound in the methanol extract. Interestingly, we discovered that the antifeedant activity was reduced when they were mixed (50:50). 相似文献
75.
Takao Morita Akihiro NezuYosuke Tojyo Akihiko Tanimura 《Biochemical and biophysical research communications》2013
Adenoviruses have been used for gene transfer to salivary gland cells in vivo. Their use to study the function of salivary acinar cells was limited by a severe inflammatory response and by the destruction of fluid-secreting acinar cells. In the present study, low doses of adenovirus were administered to express Stim1-mKO1 by retrograde ductal injection to submandibular glands. The approach succeeded in increasing muscarinic stimulation-induced Ca2+ responses in acinar cells without inflammation or decreased salivary secretions. This increased Ca2+ response was notable upon weak muscarinic stimulation and was attributed to increased Ca2+ release from internal stores and increased Ca2+ entry. The basal Ca2+ level was higher in Stim1-mKO1-expressing cells than in mKO1-expressing and non-expressing cells. Exposure of permeabilized submandibular acinar cells, where Ca2+ concentration was fixed at 50 nM, to inositol 1,4,5-trisphosphate (IP3) produced similar effects on the release of Ca2+ from stores in Stim1-mKO1-expressing and non-expressing cells. The low toxicity and relative specificity to acinar cells of the mild gene transfer method described herein are particularly useful for studying the molecular functions of salivary acinar cells in vivo, and may be applied to increase salivary secretions in experimental animals and human in future. 相似文献
76.
77.
Isoko Kuriyama Anna Miyazaki Yuko Tsuda Hiromi Yoshida Yoshiyuki Mizushina 《Bioorganic & medicinal chemistry》2013,21(2):403-411
The present study was designed to investigate the anticancer activity of novel nine small peptides (compounds 1–9) derived from TT-232, a somatostatin structural analogue, by analyzing the inhibition of mammalian DNA polymerase (pol) and human cancer cell growth. Among the compounds tested, compounds 3 [tert-butyloxycarbonyl (Boc)-Tyr-Phe-1-naphthylamide], 4 (Boc-Tyr-Ile-1-naphthylamide), 5 (Boc-Tyr-Leu-1-naphthylamide) and 6 (Boc-Tyr-Val-1-naphthylamide) containing tyrosine (Tyr) but no carboxyl groups, selectively inhibited the activity of rat pol β, which is a DNA repair-related pol. Compounds 3–6 strongly inhibited the growth of human colon carcinoma HCT116 p53+/+ cells. The influence of compounds 1–9 on HCT116 p53?/? cell growth was similar to that observed for HCT116 p53+/+ cells. These results suggest that the cancer cell growth suppression induced by these compounds might be related to their inhibition of pol. Compound 4 was the strongest inhibitor of pol β and cancer cell growth among the nine compounds tested. This compound specifically inhibited rat pol β activity, but had no effect on the other 10 mammalian pols investigated. Compound 4 combined with methyl methane sulfonate (MMS) treatment synergistically suppressed HCT116 p53?/? cell growth compared with MMS alone. This compound also induced apoptosis in HCT116 cells with or without p53. From these results, the influence of compound 4, a specific pol β inhibitor, on the relationship between DNA repair and cancer cell growth is discussed. 相似文献
78.
Masao Ohashi Takuji Oyama Endy Widya Putranto Tsuyoshi Waku Hiromi Nobusada Ken Kataoka Kenji Matsuno Masakazu Yashiro Kosuke Morikawa Nam-ho Huh Hiroyuki Miyachi 《Bioorganic & medicinal chemistry》2013,21(8):2319-2332
In the continuing study directed toward the development of peroxisome proliferator-activated receptor gamma (hPPARγ) agonist, we attempted to improve the water solubility of our previously developed hPPARγ-selective agonist 3, which is insufficiently soluble for practical use, by employing two strategies: introducing substituents to reduce its molecular planarity and decreasing its hydrophobicity via replacement of the adamantyl group with a heteroaromatic ring. The first approach proved ineffective, but the second was productive. Here, we report the design and synthesis of a series of α-benzyl phenylpropanoic acid-type hPPARγ partial agonists with improved aqueous solubility. Among them, we selected (R)-7j, which activates hPPARγ to the extent of about 65% of the maximum observed with a full agonist, for further evaluation. The ligand-binding mode and the reason for the partial-agonistic activity are discussed based on X-ray-determined structure of the complex of hPPARγ ligand-binding domain (LBD) and (R)-7j with previously reported ligand-LDB structures. Preliminal apoptotic effect of (R)-7j against human scirrhous gastric cancer cell line OCUM-2MD3 is also described. 相似文献
79.
Hiromi Toyama Eriko Hayashi Kozo Nagaoka Yujiro Yamada 《Bioscience, biotechnology, and biochemistry》2013,77(8):1859-1864
To establish a procedure for high frequency transfection in streptomycetes, the conditions and factors affecting the polyethyleneglycol (PEG) mediated transfection of S. chartreusis SF1623 by actinophage Φr5 DNA were studied. Protoplasts of S. chartreusis SF1623 prepared by treatment with lysozyme and achromopeptidase were very stable. Protoplasts from 20 to 22hr culture cells were more competent for transfection. The optimal pH of the medium for transfection was pH 7.6. The presence of NaCl, thymidine, ATP, ADP or adenosine in the transfection medium enhanced the frequency of transfection. The optimal conditions determined for protoplast transfection were 12.5% PEG 4,000, 300 mm NaCl, 1 mm thymidine, final concentration, Φr5 DNA and protoplasts in P3 medium (pH 7.6). The frequency of transfection under the optimal conditions was 5 × 105 per μg Φr5 DNA and was about 3 × 10?3 per regenerated protoplasts.Progenitively mature phages appeared 4hr after incubation in the regeneration solution and their number continued to increase for about 11 hr. The burst size was estimated to be about 400. 相似文献
80.
Junki Katsube Hiromi Shimomura Masanao Matsui 《Bioscience, biotechnology, and biochemistry》2013,77(11):1997-2004
Synthesis of several prostaglandin-F1 related compounds utilizing bicyclo(2,2,1) heptene derivatives as key intermediates were investigated. 相似文献