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   939篇
  免费   90篇
  国内免费   81篇
  1110篇
  2023年   18篇
  2022年   13篇
  2021年   26篇
  2020年   26篇
  2019年   43篇
  2018年   41篇
  2017年   41篇
  2016年   32篇
  2015年   32篇
  2014年   47篇
  2013年   56篇
  2012年   28篇
  2011年   39篇
  2010年   39篇
  2009年   50篇
  2008年   40篇
  2007年   42篇
  2006年   38篇
  2005年   35篇
  2004年   29篇
  2003年   26篇
  2002年   19篇
  2001年   23篇
  2000年   20篇
  1999年   18篇
  1998年   12篇
  1997年   9篇
  1996年   8篇
  1995年   9篇
  1994年   20篇
  1993年   10篇
  1992年   10篇
  1991年   14篇
  1990年   15篇
  1989年   12篇
  1988年   8篇
  1987年   16篇
  1986年   6篇
  1985年   5篇
  1984年   17篇
  1983年   16篇
  1982年   9篇
  1981年   12篇
  1980年   8篇
  1979年   16篇
  1978年   14篇
  1977年   12篇
  1976年   7篇
  1975年   8篇
  1974年   7篇
排序方式: 共有1110条查询结果,搜索用时 9 毫秒
111.
A method is presented that allows to combine the effective potential between two nano crystals (NC), the potential of mean force (PMF), as obtained from all-atomistic molecular dynamics simulations with perturbation theory. In this way, a functional dependence of the PMF on temperature is derived, which enables the prediction of the PMF in a wide temperature range. We applied the method to systems of capped gold NCs of different size. They show very good agreement with data from atomistic simulations.  相似文献   
112.

Background

To construct safer recombinant flavivirus vaccine, we exploited Japanese encephalitis virus (JEV) replicon-based platform to generate single-round infectious particles (SRIPs) that expressed heterologous neutralizing epitope SP70 derived from enterovirus-71 (EV71). Such pseudo-infectious virus particles, named SRIP-SP70, although are not genuine viable viruses, closely mimic live virus infection to elicit immune responses within one round of viral life cycle.

Results

We found that, besides gaining of full protection to thwart JEV lethal challenge, female outbred ICR mice, when were immunized with SRIP-SP70 by prime-boost protocol, could not only induce SP70-specific and IgG2a predominant antibodies but also provide their newborns certain degree of protection against EV71 lethal challenge.

Conclusions

Our results therefore exemplify that this vaccination strategy could indeed confer an immunized host a dual protective immunity against subsequent lethal challenge from JEV or EV71.  相似文献   
113.
Liu JJ  Wang XC  Fan B 《Bioresource technology》2011,102(9):5305-5311
The occurrence of polycyclic aromatic hydrocarbons (PAHs) in a domestic wastewater treatment plant (WWTP) was investigated in a 1 year period. In order to understand how PAHs were removed at different stages of the treatment process, adsorption experiments were conducted using quartz sand, kaolinite, and natural clay as inorganic adsorbents and activated sludge as organic adsorbent for adsorbing naphthalene, phenanthrene, and pyrene. As a result, the adsorption of PAHs by the inorganic adsorbents well followed the Langmuir isotherm while that by the activated sludge well followed the Freundlich isotherm. By bridging equilibrium partitioning coefficient with the parameters of adsorption isotherm, a set of mathematical models were developed. Under an assumption that in the primary settler PAHs removal was by adsorption onto inorganic particles and in the biological treatment unit it was by adsorption onto activated sludge, the model calculation results fairly reflected the practical condition in the WWTP.  相似文献   
114.
猪细小病毒(PPV)VP2蛋白N端连续9个甘氨酸富集的编码区是VP3蛋白的切割位点,常规PCR扩增容易导致该区段的缺失,为研究该缺失对PPV病毒样颗粒(VLPs)的影响,探索VP2病毒样颗粒上适合外源基因插入的位点,构建了该区段缺失的VP2的真核表达载体pCI-△VP2,并以完整VP2作为对照,采用脂质体介导法转染Vero细胞,通过生物信息学技术、SDS-PAGE、Western blotting、间接免疫荧光以及正染和免疫电镜对表达产物进行分析观察;进一步将重组质粒以核酸疫苗的方式直接肌注免疫小鼠,采用间接ELISA试验、淋巴细胞增殖试验和T细胞亚群流式细胞技术,分析免疫小鼠的体液和细胞免疫应答.结果显示,缺失△VP2和完整VP2在Vero细胞中均能自我装配成VLPs,并具有与完整病毒粒子类似血凝性,pCI-△VP2和pCI-VP2均可诱导小鼠产生较强的特异性体液免疫应答和良好的细胞免疫应答.结果表明,甘氨酸富集区的缺失不影响VP2病毒样颗粒的装配和免疫原性,△VP2同样可进行PPV VLPs疫苗和抗原转运载体的研制,为VLPs载体改造和修饰位点的探索提供了新方向,为VP2基因结构与蛋白质功能的关系提供了新的理论依据.  相似文献   
115.
116.
改性纳米TiO2对蓝藻的生理生态影响   总被引:1,自引:0,他引:1  
在自然光照条件和日光灯光照条件下,用掺1%Ag纳米TiO2、掺5%Fe2O3纳米TiO2、掺10%ZnO纳米TiO2和纯纳米TiO2对铜绿微囊藻(Synechocystis sp.)PCC6803和集胞藻(Microcystis aeruginosa)进行处理,并对2种藻样的叶绿素含量、类胡萝卜素含量、体外超氧自由基含量及超氧化物歧化酶(superoxidase dismutase,SOD)活性进行测定。结果表明,自然光照条件比日光灯光照条件更利于纳米材料光催化反应的进行;改性后的3种纳米材料对蓝藻的抑制效果要好于纯纳米TiO2,其中效果最好的为掺1%Ag纳米TiO2;2种藻样在适应由光催化剂引发光催化而产生的自由基胁迫时,集胞藻的适应能力比铜绿微囊藻更强。  相似文献   
117.
We describe a novel strategy to produce vaccine antigens using a plant cell‐suspension culture system in lieu of the conventional bacterial or animal cell‐culture systems. We generated transgenic cell‐suspension cultures from Nicotiana benthamiana leaves carrying wild‐type or chimeric Bamboo mosaic virus (BaMV) expression constructs encoding the viral protein 1 (VP1) epitope of foot‐and‐mouth disease virus (FMDV). Antigens accumulated to high levels in BdT38 and BdT19 transgenic cell lines co‐expressing silencing suppressor protein P38 or P19. BaMV chimeric virus particles (CVPs) were subsequently purified from the respective cell lines (1.5 and 2.1 mg CVPs/20 g fresh weight of suspended biomass, respectively), and the resulting CVPs displayed VP1 epitope on the surfaces. Guinea pigs vaccinated with purified CVPs produced humoral antibodies. This study represents an important advance in the large‐scale production of immunopeptide vaccines in a cost‐effective manner using a plant cell‐suspension culture system.  相似文献   
118.
Inflammation in vascular (mostly arterial) walls and heart valves triggered by the trans-endothelial influx of LDL particles and the action of subsequently modified (e.g., by oxidation) LDL particles can trigger true bone formation by valvar fibroblasts, by a subpopulation of re-differentiation-competent VSMCs (vascular smooth muscle cells) or by vascular pericytes. Vascular ossification can lead to heart failure and death. Elderly osteoporotic women who need osteogenic drugs to restore their lost skeletal bone are paradoxically prone to vascular ossification-the "calcification paradox." The recent introduction into the clinic of a potently osteogenic parathyroid hormone peptide, Lilly's rhPTH-(1-34)OH (Forteotrade mark), to reverse skeletal bone loss raises the question of whether this and other potently osteogenic PTHs still in clinical trial might also stimulate vascular ossification in such osteoporotic women. Indeed the VSMCs in human and rat atherosclerotic lesions hyperexpress PTHrP and the PTHR1 (or PTH1R) receptor as do maturing osteoblasts. And the evidence indicates that endogenous PTHrP with its NLS (nuclear/nucleolar localization sequence) does stimulate VSMC proliferation (a prime prerequisite for atheroma formation and ossification) via intranuclear targets that inactivate pRb, the inhibitory G1/S checkpoint regulator, by stimulating its hyperphosphorylation. But neither externally added full-length PTHrP nor the NLS-lacking PTHrP-(1-34)OH gets into the VSMC nucleus and instead they inhibit proliferation and calcification by only activating the cell's PTHR1 receptors. No PTH has an NLS and, as expected from the observations on the externally added PTHrPs, hPTH-(1-34)OH inhibits calcification by VSMCs and cannot stimulate vascular ossification in a diabetic mouse model. Encouraging though this may be for osteoporotics with their "calcification paradox," more work is needed to be sure that the skeletally osteogenic PTHs do not promote vascular ossification with its cardiovascular consequences.  相似文献   
119.
An internally truncated C gene of adr hepatitis B virus core antigen with long internal deletion (aa81–aa116) (ΔHBcAg with 36aa truncation) was expressed in Saccharomyces cerevisiae and the products (ΔrHBcAg) were purified from a crude lysate of the yeast by three steps: Sephrose CL-4B chromatography, sucrose step-gradient ultracentrifugation and CsCl-isopycnic ultracentrifugation. Results of ELISA test and density analysis of CsCl-isopycnic ultracentrifugation indicated that the purified products (ΔrHBcAg protein) with HBeAg antigenicity mainly located at the densities of 1.23 g ml−1. Observation and analysis of the purified ΔrHBcAg products by AFM indicated that the ΔrHBcAg (core) protein produced in S. cerevisiae could self-assemble into three or more size classes of core particles which exhibited a polymorphous distribution of ΔrHBcAg (core) particles. These different size classes of core particles mainly centred on the range whose mean diameter was from 10 nm to 48 nm, especially on the position of 11 nm, 15.6 nm and the range from 27 nm to 41 nm, respectively. Furthermore, the most number of core particles mainly centred on the range whose mean diameter was from 27 nm to 41 nm. These results above indicated that the truncated internal long fragment (aa81–aa116) probably had no effect on self-assembly of the HBcAg core particles which implied the internal length fragment (aa81–aa116) was not the sole domain for self-assembly of HBcAg dimer or the truncated HBcAg protein subunit formed the fresh interactive domain with each other. These initial results above by AFM analysis were very important for further research on the self-assembly, ultrastructure, subunit interaction and core internal deletion mutant (CIDM) function of HBcAg core particles.  相似文献   
120.
Diesel exhaust particles (DEP) are a major source of air-borne pollution and are linked to increased risk of disease including lung cancer. Here we investigated effects of exposure to DEP on the frequency of DNA deletions, levels of oxidative DNA damage and DNA adduct formation during embryonic development in mice. Pregnant dams were orally exposed to various doses of DEP (500, 250, 125, 62.5, 31.25 mg/kg/day) at embryonic days 10.5–15.5. We determined the frequency of 70 kb DNA deletions spanning exons 6–18 at the pun allele that results in black-pigmented spots in the unpigmented retinal pigment epithelium in the eyes of pun/pun offspring mice. DEP caused a significant increase in the frequency of DNA deletions. Levels of 8-OH deoxyguanosine indicating oxidative DNA damage were within the limits of the unexposed mouse embryos. 33P post-labeling analysis revealed very low levels of DNA adducts in the embryo tissue. Thus, transplacental exposure to DEP resulted in a significant increase in the frequency of DNA deletions in the mouse fetus and such genetic alterations in the offspring may have pathological consequences later in life.  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