全文获取类型
收费全文 | 1610篇 |
免费 | 70篇 |
国内免费 | 112篇 |
专业分类
1792篇 |
出版年
2024年 | 1篇 |
2023年 | 24篇 |
2022年 | 43篇 |
2021年 | 81篇 |
2020年 | 54篇 |
2019年 | 65篇 |
2018年 | 53篇 |
2017年 | 35篇 |
2016年 | 57篇 |
2015年 | 91篇 |
2014年 | 100篇 |
2013年 | 103篇 |
2012年 | 161篇 |
2011年 | 131篇 |
2010年 | 75篇 |
2009年 | 70篇 |
2008年 | 96篇 |
2007年 | 75篇 |
2006年 | 73篇 |
2005年 | 56篇 |
2004年 | 56篇 |
2003年 | 40篇 |
2002年 | 21篇 |
2001年 | 38篇 |
2000年 | 21篇 |
1999年 | 20篇 |
1998年 | 15篇 |
1997年 | 17篇 |
1996年 | 13篇 |
1995年 | 10篇 |
1994年 | 11篇 |
1993年 | 12篇 |
1992年 | 18篇 |
1991年 | 12篇 |
1990年 | 10篇 |
1989年 | 6篇 |
1988年 | 7篇 |
1987年 | 5篇 |
1986年 | 6篇 |
1985年 | 3篇 |
1984年 | 1篇 |
1983年 | 4篇 |
1981年 | 1篇 |
1980年 | 1篇 |
排序方式: 共有1792条查询结果,搜索用时 15 毫秒
91.
Effects of microclimate,litter type,and mesh size on leaf litter decomposition along an elevation gradient in the Wuyi Mountains,China 总被引:1,自引:0,他引:1
Litter decomposition is an important ecosystem process regulated by both biotic factors (e.g., decomposers and litter types) and abiotic factors (e.g., temperature and moisture). This study examined the regulatory effects of soil fauna and microclimate on decomposition of two substrates (Castanopsis carlesii and Pinus taiwanensis) along an elevation gradient in four ecosystems of zonal vegetation types in southeastern China: evergreen broadleaf forest (EVB), coniferous forest (COF), dwarf forest (DWF), and alpine meadow (ALM). Our objective was to identify the mechanisms by which microclimate, substrate, and fauna control litter decomposition, especially where variations in ecosystem structure and environment are markedly shown across an elevation gradient. The hypotheses were as follows: (1) litter decomposition within the same litter type would decrease across the elevation gradient, (2) litter decomposition would be lower in poorer nutrient quality substrate across the four sites, and (3) litter dynamics, influenced by strong interactions among ecosystem type, litter type, and decomposers, would vary by elevation gradient due to microclimate effects (i.e., temperature and moisture). The decomposition rates of C. carlesii were significantly higher than those of P. taiwanensis at EVB, COF, and DWF sites; however, they were not significantly different at the ALM site. Low elevation forests possessed a microclimate (warm and humid) that favors decomposer activities and also appeared to possess a decomposer community adapted to consuming large amounts of leaf litter, as indicated by the rapid leaf litter loss. Litter decomposition in micro-mesh bags proceeded more slowly compared to litter in meso-mesh and macro-mesh litterbags across the elevation gradient, indicating that restricting some detritivore access to litter reduced litter mass loss. We suggest that microclimate and faunal contributions to plant litter decomposition differ markedly across the ecosystems in the Wuyi Mountains. 相似文献
92.
Functional expression and purification of bovine enterokinase light chain in recombinant Escherichia coli 总被引:2,自引:0,他引:2
Enterokinase (EC 3.4.21.9) is a serine proteinase of the intestinal brush border that exhibits specificity for the sequence (Asp)(4)-Lys and converts trypsinogen into its active form, trypsin. A codon optimized sequence coding light chain (catalytic subunit) of bovine enterokinase gene (sBEKLC) was synthesized, and it was fused with DsbA to construct the expression vector (pET39-sBEKLC). Then, the plasmid was transformed into E. coli BL21 (DE3) for expression. Under optimal conditions, the volumetric productivity of fusion protein reached 151.2 mg L(-1), i.e., 80.6 mg sBEKLC L(-1). The cold osmotic shock technique was successfully used to extract sBEKLC from periplasmic space, and nickel affinity chromatography was employed to obtain mature sBEKLC. Finally, about 6.8 mg of bioactive sBEKLC was purified from 1 liter fermentation broth and could be used to cleave one tested fusion protein with an inter-domain enteropeptidase recognition site. This work will be helpful for large-scale production of this increasingly demanded enterokinase. 相似文献
93.
附子多糖FI的分离纯化及部分理化性质研究 总被引:3,自引:0,他引:3
白附片经热水抽提、Sevag法脱蛋白、乙醇沉淀、DEAE- C32柱层析分离 ,再通过 Sephadex G- 2 0 0柱层析进一步纯化 ,得到一种纯白色粉末状多糖 ,糖含量为 97% ,平均分子量为 2 .6× 10 5,熔点为 2 70℃。经完全酸水解、薄层层析、红外光谱分析 ,证明为葡聚糖。 相似文献
94.
95.
不同佐剂对含S+PreS1融合抗原的乙型肝炎病毒颗粒疫苗免疫原性的影响 总被引:2,自引:1,他引:2
本研究在前期工作基础上,用CHO细胞表达的含PreS1+S融合抗原的新型基因工程HBV颗粒疫苗(HBSS1)与Al(OH)3、CpG及CpG+Al(OH)3等佐剂配伍,在Balb/C小鼠模型上研究不同佐剂对HBV颗粒疫苗肌肉注射后免疫应答的影响,主要包括抗体滴度、抗体亚型分类及特异性细胞免疫(γ-IFNELISpot检测)。结果表明:CpG佐剂结合HBSS1颗粒疫苗可快速诱导(单针免疫)高水平的抗PreS1及S抗体,IgG2a/IgG1比率1,同时可诱导较高抗原特异的细胞免疫应答;Al(OH)3+CpG双佐剂组一次免疫后可诱导产生最高的抗S抗体滴度(1:105),其产生的抗体亚类包括IgG1、IgG2a与IgG2b;在S抗原N端(13~49aa)存在优势CTL表位。结论:CpG佐剂结合HBSS1颗粒疫苗应是发展新型治疗性乙肝疫苗的较佳选项。 相似文献
96.
Wen Yang Jing Liu Fanfan Zheng Meixiang Jia Linnan Zhao Tianlan Lu Yanyan Ruan Jishui Zhang Weihua Yue Dai Zhang Lifang Wang 《PloS one》2013,8(4)
Background
Autism is a neurodevelopmental disorder with a high estimated heritability. ATP2B2, located on human chromosome 3p25.3, encodes the plasma membrane calcium-transporting ATPase 2 which extrudes Ca2+ from cytosol into extracellular space. Recent studies reported association between ATP2B2 and autism in samples from Autism Genetic Resource Exchange (AGRE) and Italy. In this study, we investigated whether ATP2B2 polymorphisms were associated with autism in Chinese Han population.Methods
We performed a family based association study between five SNPs (rs35678 in exon, rs241509, rs3774180, rs3774179, and rs2278556 in introns) in ATP2B2 and autism in 427 autism trios of Han Chinese descent. All SNPs were genotyped using the Sequenom genotyping platform. The family-based association test (FBAT) program was used to perform association test for SNPs and haplotype analyses.Results
This study demonstrated a preferential transmission of T allele of rs3774179 to affected offsprings under an additive model (T>C, Z = 2.482, p = 0.013). While C allele of rs3774179 showed an undertransmission from parents to affected children under an additive and a dominant model, respectively (Z = −2.482, p = 0.013; Z = −2.591, p = 0.0096). Haplotype analyses revealed that three haplotypes were significantly associated with autism. The haplotype C-C (rs3774180–rs3774179) showed a significant undertransmission from parents to affected offsprings both in specific and global haplotype FBAT (Z = −2.037, p = 0.042; Global p = 0.03). As for the haplotype constructed by rs3774179 and rs2278556, C-A might be a protective haplotype (Z = −2.206, p = 0.027; Global p = 0.04), while T-A demonstrated an excess transmission from parents to affected offsprings (Z = 2.143, p = 0.032). These results were still significant after using the permutation method to obtain empirical p values.Conclusions
Our research suggested that ATP2B2 might play a role in the etiology of autism in Chinese Han population. 相似文献97.
98.
[目的]改造谷氨酸棒杆菌(Corynebacterium glutamicum)中NADPH合成途径,阻断胞内NADPH的合成,获得1株NADPH营养缺陷型菌株。[方法]通过失活L-赖氨酸高产菌C. glutamicum Lys-χ中葡萄糖-6-磷酸脱氢酶(Zwf)和苹果酸酶(MalE)并将NADP~+依赖型异柠檬酸脱氢酶(NADP~+-Icdcg)替换成变形链球菌(Streptococcus mutans)中的NAD~+-Icdsm,阻断胞内NADPH的合成。随后结合辅因子工程,引入大肠杆菌(Escherichia coli)中膜结合吡啶核苷酸转氢酶(PntAB)并通过不同强度启动子控制PntAB的表达水平。最后,分析不同重组菌中胞内氧化还原水平和L-赖氨酸生产强度的变化。[结果]重组菌C.glutamicum Lys-χΔZMI_(Cg)::I_(Sm)(即Lys-x1)胞内检测不到NADPH,为1株NADPH营养缺陷型菌株。该重组菌只在以葡萄糖酸为碳源的基础培养基中生长和积累L-赖氨酸,而以葡萄糖、丙酮酸、α-酮戊二酸和草酰乙酸为碳源时无法生长。此外,表达E.coli中的PntAB可回补重组菌Lys-χ1胞内NADPH的水平,但由于不同强度启动子控制PntAB表达水平不同,重组菌胞内NADPH水平也不同,并影响L-赖氨酸的生产强度。[结论]重组菌Lys-χ1可作为有效的底盘细胞,用于考察不同的NADPH再生策略,获得不同胞内NADPH水平的重组菌株,为进一步阐明NADPH调控微生物细胞生理代谢功能的机制提供研究基础。 相似文献
99.
100.
探讨建立一种适合贵州地区、高效、稳定的小鼠超数排卵优化方案。在饲养环境相同的基础上,对激素(PMSG, hCG)不同的剂量组合、注射间隔时间、小鼠周龄等影响因素进行了相关研究。试验结果表明:(1)平均采胚数量组间、平均异常胚组间与平均可用胚组间差异显著(p<0.05),注射10 IU的激素剂量组合获得受精卵最多,且异常胚最少,效果最佳。(2)第1、第2、第3组平均采胚数量组间差异显著(p<0.05),第1组与第2组平均可用胚组间差异不显著(p>0.05),但第1、2组与第3组差异显著(p<0.05),异常胚组间差异不显著(p>0.05),选择4周龄超排效果最佳。(3)第1、第2、第3组平均采胚数量、平均可用胚组间差异显著(p<0.05),平均异常胚组间差异不显著(p>0.05),PMSG,hCG间隔注射时间为48 h为最佳。 相似文献