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   22180篇
  免费   1525篇
  国内免费   988篇
  24693篇
  2024年   54篇
  2023年   295篇
  2022年   632篇
  2021年   1015篇
  2020年   675篇
  2019年   901篇
  2018年   891篇
  2017年   679篇
  2016年   958篇
  2015年   1336篇
  2014年   1511篇
  2013年   1753篇
  2012年   1950篇
  2011年   1819篇
  2010年   1016篇
  2009年   907篇
  2008年   1012篇
  2007年   928篇
  2006年   845篇
  2005年   725篇
  2004年   632篇
  2003年   526篇
  2002年   475篇
  2001年   309篇
  2000年   320篇
  1999年   282篇
  1998年   174篇
  1997年   161篇
  1996年   181篇
  1995年   162篇
  1994年   136篇
  1993年   102篇
  1992年   151篇
  1991年   150篇
  1990年   124篇
  1989年   97篇
  1988年   98篇
  1987年   101篇
  1986年   70篇
  1985年   89篇
  1984年   50篇
  1983年   53篇
  1982年   27篇
  1981年   26篇
  1980年   24篇
  1979年   35篇
  1978年   28篇
  1977年   20篇
  1975年   29篇
  1974年   21篇
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
101.
朱俊真 《遗传学报》1992,19(2):101-106
应用人X染色体α卫星DNA探针进行X染色体正常或异常个体的外周血淋巴细胞染色体和间期核的原位杂交,在R显带的中期分裂相上,绝大部分杂交颂粒位于X染色体着丝粒区(p11→q11);在间期核内则显现与X染色体数相一致的银颗粒簇,其中相当部分位于核边缘区。实验结果表明,用原位杂交来检测X染色体数目,比记数Barr小体的方法可靠。本文还就α卫星DNA探针在间期细胞遗传学方面广泛的应用做了讨论。  相似文献   
102.
本文给出保证传染病模型非线性积分方积x(t)=integral from n=t-τto t f[s,x(s)]ds的非零周期解存在唯一的一组充分条件。  相似文献   
103.
104.
Yang Y  Ma J  Song Z  Wu M 《FEBS letters》2002,532(1-2):36-44
Several novel prokaryotic and eukaryotic expression vectors were constructed for protein transduction and subcellular localization. These vectors employed an N-terminal stretch of 11 basic amino acid residues (47-57) from the human immunodeficiency virus type 1 (HIV-1) TAT protein transduction domain (PTD) for protein translocation and cellular localization. The vectors also contained a six-histidine (His(6)) tag at the N- or C-terminus for convenient purification and detection, and a multiple cloning site for easy insertion of foreign genes. Some heterologous genes including HSV-TK, Bcl-rambo, Smac/DIABLO and GFP were fused in-frame to TAT PTD and successfully overexpressed in Escherichia coli. The purified TAT-GFP fusion protein was able to transduce into the mammalian cells and was found to locate mainly in the cytosol when exogenously added to the cell culture medium. However, using a transfection system, mammalian-expressed TAT-GFP predominantly displayed a nuclear localization and nucleolar accumulation in mammalian cell lines. This discrepancy implies that the exact subcellular localization of transduced protein may depend on cell type, the nature of imported proteins and delivery approach. Taken together, our results demonstrate that a TAT PTD length of 11 amino acids was sufficient to confer protein internalization and its subsequent cellular localization. These novel properties allow these vectors to be useful for studying protein transduction and nuclear import.  相似文献   
105.
Magnetic hot spots, which implies confinements and enhancements of magnetic fields, are demonstrated in graphene junctions (GJs) in the mid-infrared range. The appearance of magnetic hot spots in GJs comes from the conduction currents in the junction. In further, the extinction resonance peaks suffer blue shift, along with the increases in the magnetic fields inside junction area, when the junction width reduces. In opposite to the circumstances for electric field enhancements, neither magnetic field enhancements nor resonance frequency of GJs is perturbed by the intrinsic nonlocal electronic response of graphene. Such nonlocality immunized magnetic enhancement could be explained by the polarization dependent property of nonlocal effect.  相似文献   
106.
壳聚糖固定化AS1.398中性蛋白酶稳定性的研究   总被引:3,自引:0,他引:3  
对以壳聚糖为载体,由戊二醛作交联剂。制备的固定化AS1.398中性蛋白酶的稳定性进行了研究。实验结果表明,固定化AS1.398中性蛋白酶对热、乙醇、尿素以及pH值的稳定性均有明显的提高。  相似文献   
107.

Background

Fungal rhinosinusitis has become an increasingly recognized disease, being Aspergillus species responsible for most of the cases. Its diagnosis is quite difficult because of the non-specific symptoms and low sensitivity of the current diagnostic methods.

Aims

An Aspergillus-specific nested polymerase chain reaction (PCR) assay using biopsy specimens taken from the maxillary sinuses was performed in order to assess its usefulness. Conventional diagnostic methods (histology and culture) were also carried out.

Methods

A case–control study was performed in the Institute of Stomatology, Jagiellonian University in Kraków, between 2011 and 2014. The case group consisted of 21 patients with suspected rhinosinusal mycetoma while the control group included 46 patients with no suspicion of fungal rhinosinusitis. The two-step PCR assay amplified an Aspergillus specific portion of the 18S rRNA gene. Interval estimation of sensitivity, specificity, positive (PPV) and negative (NPV) predictive values were calculated to assess the diagnostic test performance. The agreement between the PCR and the other tests was evaluated using the Kappa coefficient (k).

Results

Ninety percent of the samples obtained from patients diagnosed with mycetoma yielded positive PCR results. The PCR showed almost perfect concordance with histology (k = 0.88). Sensitivity, specificity, PPV and NPV estimates were 90%; 95% CI: (55.5–99.7%), 98.3%; 95% CI: (90.9–100%), 90%; 95% CI: (55.5–99.7%) and 98.3%; 95% CI: (90.9–100%), respectively. One clinical sample showed growth of Aspergillus fumigatus and positive PCR despite the negative histological examination.

Conclusions

Nested PCR assay is a promising diagnostic tool to evaluate the presence of Aspergillus in the tissue of maxillary sinus from patients with suspicion of sinus aspergillosis.  相似文献   
108.
Agrobacterium rhizogenes-mediated genetic transformation of Saussurea involucrata was investigated. Four bacterial strains, A4, LBA 9402, R1000 and R1601 and three explant types, leaf blade, petiole and root, were examined. Over 100 hairy root lines were successfully established with strains R1601, R1000 and LBA9402, but none with A4. The highest transformation efficiency of 67% was achieved by using strain R1601 with root explants. One hairy root line isolated from this combination, HR1601-1, produced up to 43.5 ± 1.13 mg syringin g−1 dw, which is about 50-fold higher than that in the wild type plants.Two other lines, HR1000-1 and HRLBA9402-1, isolated from R1000- and LBA9402-transformed roots, respectively, also displayed high capacity of syringin production, being 32.5 ± 3.08 and 39.7 ± 1.37 mg syringin g−1 dw. These three lines were characterized in detail. Polymerase chain reaction analyses confirmed these root lines were of A. rhizogenes origin.  相似文献   
109.
Mutations in polycystin-1 (PC1) can cause autosomal dominant polycystic kidney disease, which is a leading cause of renal failure. The available evidence suggests that PC1 acts as a mechanosensor, receiving signals from the primary cilia, neighboring cells, and extracellular matrix. PC1 is a large membrane protein that has a long N-terminal extracellular region (about 3000 amino acids) with a multimodular structure including 16 Ig-like polycystic kidney disease (PKD) domains, which are targeted by many naturally occurring missense mutations. Nothing is known about the effects of these mutations on the biophysical properties of PKD domains. Here we investigate the effects of several naturally occurring mutations on the mechanical stability of the first PKD domain of human PC1 (HuPKDd1). We found that several missense mutations alter the mechanical unfolding pathways of HuPKDd1, resulting in distinct mechanical phenotypes. Moreover, we found that these mutations also alter the thermodynamic stability of a structurally homologous archaeal PKD domain. Based on these findings, we hypothesize that missense mutations may cause autosomal dominant polycystic kidney disease by altering the stability of the PC1 ectodomain, thereby perturbing its ability to sense mechanical signals.  相似文献   
110.
N-乙酰氨基葡萄糖化在信号转导中的作用   总被引:2,自引:0,他引:2  
Li JL  Ma L 《生理科学进展》2002,33(4):373-375
蛋白质磷酸化在生命活动以及信号转导过程中的重要作用已经被研究证实,但不少研究发现在大多数核,胞液蛋白质上不仅存在磷酸化动态修饰,还存在广泛的动态N-乙酰氨基葡萄糖修饰,N-乙酰氨基葡萄糖基转移酶和N-乙酰氨基葡萄糖基酶以类似于蛋白质激酶和磷酸酶的方式调节蛋白质是否发生N-乙酰氨基葡萄糖化。N-乙酰氨基葡萄糖化蛋白质主要分布在细胞核与胞液,其生理功能涉及细胞基本生命活动和调节信号传递。N-乙酰氨基葡萄糖的作用基础与阻断或影响蛋白质的磷酸化有关。  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