首页 | 本学科首页   官方微博 | 高级检索  
文章检索
  按 检索   检索词:      
出版年份:   被引次数:   他引次数: 提示:输入*表示无穷大
  收费全文   257篇
  免费   20篇
  国内免费   43篇
  2024年   2篇
  2022年   4篇
  2021年   8篇
  2020年   3篇
  2019年   3篇
  2018年   2篇
  2017年   8篇
  2016年   8篇
  2015年   9篇
  2014年   14篇
  2013年   12篇
  2012年   13篇
  2011年   16篇
  2010年   11篇
  2009年   13篇
  2008年   15篇
  2007年   17篇
  2006年   19篇
  2005年   14篇
  2004年   10篇
  2003年   9篇
  2002年   10篇
  2001年   8篇
  2000年   3篇
  1999年   5篇
  1998年   5篇
  1997年   3篇
  1996年   2篇
  1995年   2篇
  1993年   2篇
  1992年   9篇
  1991年   9篇
  1990年   6篇
  1989年   7篇
  1988年   2篇
  1987年   2篇
  1986年   4篇
  1985年   2篇
  1984年   2篇
  1983年   6篇
  1980年   4篇
  1979年   4篇
  1978年   1篇
  1977年   1篇
  1976年   1篇
  1973年   2篇
  1970年   1篇
  1969年   1篇
  1967年   1篇
  1962年   1篇
排序方式: 共有320条查询结果,搜索用时 15 毫秒
61.
依据1996年、2003—2007年扎龙保护区丹顶鹤巢址数据,利用Voronoi图理论,分析了巢址的空间分布特征,计算了其拓扑指数—Voronoi图面积。结果表明,丹顶鹤巢址空间分布具有明显的空间聚类特征。以扎龙保护区的2006年巢址、居民点、国道和铁路为对象,构成其Voronoi图,较好地说明了人类活动对丹顶鹤巢址的空间格局的影响,研究结果对丹顶鹤栖息地的保护将起到重要作用。  相似文献   
62.
Urinary incontinence can occur poststroke owing to weakness or incoordination of sphincter muscles, impaired bladder sensation, or hyperreflexic, neurogenic bladder. Four male subjects who had urinary incontinence associated with a stroke that had occurred 8 months to 10 years earlier, and who averaged 1.6 to 7.5 accidental voidings per week, participated in an outpatient study with a 4-week scheduled-voiding baseline, 2 to 5 sessions of biofeedback-assisted bladder retraining, and 6- to 12-month follow-up. Training sessions included stepwise filling of the bladder and manometric feedback display of bladder pressure, abdominal pressure, and external anal sphincter pressure. Training procedures were designed to teach subjects to attend to bladder sensations, inhibit bladder contractions, and improve voluntary sphincter muscle control. All four subjects achieved and maintained continence regardless of substantial differences in subject characteristics, including laterality of stroke, degree of sensory impairment, and independence in daily activities.  相似文献   
63.
A chemical toxicity and teratogenicity test was adapted to assess potential adverse effects of a microbial pest control agent on a nontarget fish. Developing embryos of the inland silverside, Menidia beryllina, were exposed to conidiospores of the insect-pathogenic fungus Beauveria bassiana. Embryo rupture and death were observed. Embryo rupture did not always result in death, nor was death always associated with embryo rupture. Adherence of spores to the chorion, followed by germination and penetration by the germ tube, probably caused the embryos to rupture. Statistically significant (P less than or equal to 0.05) responses were observed in tests in which conidiospore concentrations were greater than or equal to 8.3 x 10(4) or less than or equal to 1.5 x 10(6)/ml. Conidiospores treated with a dispersant (biological detergent) showed significantly less binding (P less than or equal to 0.01) to embryos than did untreated spores. Both detergent-treated and heat-killed spores failed to cause significant adverse effects.  相似文献   
64.
This paper probes into the feasibility of increasing expression level of hFIX gene with endogenous intron 1 sequence.hFIX minigene was obtained with middle sequence truncated intron 1 inserted into the relative site of hFIX cDNA,and plasmid vector pKG5i‘IX,retroviral vector G1NaCi‘IX were constructed.These vectors were transduced into target cells of PA317,C2C12,primary rabbit skin fibroblasts (RSF) and primary human skin fibroblasts (HSF).The expression level of mixed colonies are PA317/pKG5i‘IX,151 ng/10^6 cells/24h;PA317/G1NaCi‘IX,308 ng/10^6 cells/24 h;C2C12/G1NaCi‘IX,186 ng/10^6 cells/24 h;RSF/G1NaCi‘IX,1929 ng/10^6 cells/24 h;HSF/G1NaCi‘IX,1646 ng/10^6 cells/24 h.These results indicated that hFIX minigene with intron l is able to increase the expression level to about 3 times of that of hFIX cDNA.Meanwhile,in order to study the application of hFIX minigene in the retroviral-mediated gene transfer system and refrain from intron splicing during viral production,a retroviral vector G1NaCi‘IXR with reversely inserted hFIX minigene expression cassette was constructed.The expression level of reverse constructor in PA317 cells was 390 ng/10^6 cells/24 h with 79% of bioactivity.PCR detection of HT/G1NaCi‘IXR cells infected with PA317/G1NaCi‘IXR supernatant confirmed the existence of intron 1 sequence.These results suggested that expression vector with forward-inserted intronl-carrying hFIX expression cassette can be used in directed gene transfer,but when using the retroviral-mediated gene transfer system,reversely-inserted intronl-carrying hFIX expression cassette should be considered.  相似文献   
65.
Gram‐negative pathogens often use conserved type three secretion systems (T3SS) for virulence. The Shigella type three secretion apparatus (T3SA) penetrates the host cell membrane and provides a unidirectional conduit for injection of effectors into host cells. The protein Spa47 localizes to the base of the apparatus and is speculated to be an ATPase that provides the energy for T3SA formation and secretion. Here, we developed an expression and purification protocol, producing active Spa47 and providing the first direct evidence that Spa47 is a bona fide ATPase. Additionally, size exclusion chromatography and analytical ultracentrifugation identified multiple oligomeric species of Spa47 with the largest greater than 8 fold more active for ATP hydrolysis than the monomer. An ATPase inactive Spa47 point mutant was then engineered by targeting a conserved Lysine within the predicted Walker A motif of Spa47. Interestingly, the mutant maintained a similar oligomerization pattern as active Spa47, but was unable to restore invasion phenotype when used to complement a spa47 null S. flexneri strain. Together, these results identify Spa47 as a Shigella T3SS ATPase and suggest that its activity is linked to oligomerization, perhaps as a regulatory mechanism as seen in some related pathogens. Additionally, Spa47 catalyzed ATP hydrolysis appears to be essential for host cell invasion, providing a strong platform for additional studies dissecting its role in virulence and providing an attractive target for anti‐infective agents.  相似文献   
66.
从康氐木霉(Trichoderma k(?)ningii)白色变异株As 3.4001的粗酶制剂中,获得了纤维素酶系中的一组C_x酶(C_(x1) C_(x2) C_(x3) C_(x4))。分离步骤包括Sephadex G-75凝胶过滤,DEAESephadex A-50离子交换层析,ConA-Sepharose亲合层析,SE-Sephadex C-50离子交换层析及聚丙烯酰胺凝胶电泳。C_(x1)与C_(x2)的分子量不同而所带电荷相同,它们的分子量各自为44,500和34,000。C_(x2)—C_(x4)的分子量相同而所带电荷不同。纯化的C_(x1)—C_(x4)经聚丙烯酰胺凝胶电泳鉴定为单带。比较它们对羧甲基纤维素钠(CMC-Na)的糖化力及液化力表明在作用方式的随机性上C_(x2)>C_(x3)>C_(x1)>C_(x4)。  相似文献   
67.
Both Staphylococcus aureus strains 243 and S-6 cells producing enterotoxin B and free enterotoxin in food and culture medium were rapidly demonstrated by using the fluorescent-antibody technique. Comparison of cell fluorescence and enterotoxin B production determined by double gel diffusion showed that an estimation of enterotoxin production could be made by observing the degree of cell fluorescence. The fluorescent-antibody technique was used to determine whether cells were producing enterotoxin under varying nutritional and environmental conditions: NaCl concentration, culture aeration, and time and temperature of incubation in Brain Heart Infusion broth and shrimp slurries. At the various NaCl concentrations, the fluorescence of cells was found positively associated with enterotoxin B production only during the first 12 hr of growth. As the NaCl concentration was increased from 0 to 10%, the fluorescence of cells and toxin production decreased. Maximum for cell fluorescence and enterotoxin production was observed at 37 C. Little or no difference in cell fluorescence and enterotoxin production with both strains was found between Brain Heart Infusion broth and shrimp slurry cultures. All results obtained with the fluorescent-antibody technique were verified with double gel diffusion for enterotoxin detection and quantitation.  相似文献   
68.
This investigation was conducted to characterize the algal inhibitory material originally isolated by Proctor (1957b) from Chlamydomonas. Analysis of purified extracts demonstrated that the inhibitor was a mixture of fatty acids. The identity and relative percentages of most of the components of the mixture were determined by GLC analysis. There is evidence that unsaturated fatty acids were responsible for the majority of the toxicity.  相似文献   
69.
Homoserine has been analyzed quantitatively by gas-liquid chromatography of its N-heptafluorobutyryl isopropyl ester. The method was confirmed by analysis of the soluble amino acid fraction of pea leaves. The possible use of the method for analysis of methionine is discussed.  相似文献   
70.
The effect of surface adsorption on the structure and stability of proteins is a matter of increasing interest in biotechnology. Therefore, we have examined the effect of adsorption to silica on the thermal stability of 7 proteins employing differential scanning calorimetry (DSC) and front surface fluorescence (FSF) spectroscopy. In general, it was found that surface adsorption decreased the thermal stability of the bound protein. Using lysozyme for further studies, DSC, FSF, and FTIR spectroscopies, as well as enzymatic activity measurements, were used to explore the effect of decreasing surface apolarity on stability. It was observed that increasing surface apolarity produced decreasing stability and increasing structural alteration of the adsorbed protein.  相似文献   
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号