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251.
The Tn5-sacB-labeled symbiotic megaplasmid pMhHN3015c of Mesorhizobium huakuii HN3015 was, respectively, transferred into M. huakuii HN308SR containing three large plasmids of pMhHN308a, pMhHN308b and pMhHN308c, and 7653R-1SR, a symbiotic plasmid pMh7653Rb deleted mutant from M. huakuii 7653R by tri-parent mating. The stable indigenous plasmid pMhHN308c of HN308SR was cured by the introduction of pMhHN3015c and the transconjugant was named as HN308SRN18. The results implied that pMhHN3015c and pMhHN308c were incompatible and might be ascribed to the same incompatibility group. Furthermore, the results from plasmid curing tests of HN308SRN18 containing pMhHN3015c, pMhHN308b, and pMhHN308a showed that not only was pMhHN3015c deleted, but that pMhHN308a was also cured simultaneously. The plasmid profiles of transconjugant 7653R-1SRN18 showed pMhHN3015c could coexist with pMh7563Ra. The plasmid replication repC-like gene sequences were detected by polymerase chain reaction from 7653R-1SRN18, HN308SRN18 and its plasmid-curing derivatives, but failed to detect from plasmid-curing derivatives of 7653R-1SRN18. The repC gene sequence similarities of strains tested were up to 99%. Results from plant nodulation tests showed that introduction of pMhHN3015c failed to restore the nitrogen fixation ability of HN308SRN18 and 7653R-1SRN18.  相似文献   
252.
海南岛位于我国南部, 地处热带北缘, 其独特的岛屿气候环境孕育了丰富的生物资源, 为我国生物多样性热点地区之一。为探究岛内的翼手目物种多样性状况, 本研究组使用雾网、蝙蝠竖琴网等工具, 于2002年至2016年先后对海南岛进行了15次翼手目多样性调查, 并根据其外形与头骨特征及系统发育学方法进行标本鉴定。共获取了1,025号标本, 隶属5科15属31种, 其中2016年12月21日在海南琼中捕获的艾氏管鼻蝠(Murina eleryi)为海南岛蝙蝠分布新记录。结合前人调查及发表结果统计, 岛内共有翼手类8科20属41种。同时基于本调查采集位点和前人调查位置信息(共计363个位点), 结合WorldClim 32种气候数据, 运用最大熵模型(MaxEnt)对海南岛翼手目物种的分布进行预测, 结果显示五指山、吊罗山、鹦哥岭、尖峰岭及海口火山口国家地质公园等地为翼手目物种多样性较丰富的区域, 而三亚、澄迈、屯昌、临高、琼海等地翼手目物种多样性较低。本研究结果为海南岛翼手目资源分布及多样性状况提供了基础资料, 也为岛内后续开展翼手目资源保护管理、蝙蝠疾病防控等提供了重要的参考依据。  相似文献   
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255.
The reproducibility of conventional two-dimensional (2D) gel electrophoresis can be improved using differential in-gel electrophoresis (DIGE), a new emerging technology for proteomic analysis. In DIGE, two pools of proteins are labeled with 1-(5-carboxypentyl)-1'-propylindocarbocyanine halide (Cy3) N-hydroxy-succinimidyl ester and 1-(5-carboxypentyl)-1'-methylindodi-carbocyanine halide (Cy5) N-hydroxysuccinimidyl ester fluorescent dyes, respectively. The labeled proteins are mixed and separated in the same 2D gel. 2D DIGE was applied to quantify the differences in protein expression between laser capture microdissection-procured esophageal carcinoma cells and normal epithelial cells and to define cancer-specific and normal-specific protein markers. Analysis of the 2D images from protein lysates of approximately 250,000 cancer cells and normal cells identified 1038 protein spots in cancer cell lysates and 1088 protein spots in normal cell lysates. Of the detected proteins, 58 spots were up-regulated by >3-fold and 107 were down-regulated by >3-fold in cancer cells. In addition to previously identified down-regulated protein annexin I, tumor rejection antigen (gp96) was found up-regulated in esophageal squamous cell cancer. Global quantification of protein expression between laser capture-microdissected patient-matched cancer cells and normal cells using 2D DIGE in combination with mass spectrometry is a powerful tool for the molecular characterization of cancer progression and identification of cancer-specific protein markers.  相似文献   
256.
Yi L  Li Z  Yuan K  Qu X  Chen J  Wang G  Zhang H  Luo H  Zhu L  Jiang P  Chen L  Shen Y  Luo M  Zuo G  Hu J  Duan D  Nie Y  Shi X  Wang W  Han Y  Li T  Liu Y  Ding M  Deng H  Xu X 《Journal of virology》2004,78(20):11334-11339
Severe acute respiratory syndrome coronavirus (SARS-CoV) is the pathogen of SARS, which caused a global panic in 2003. We describe here the screening of Chinese herbal medicine-based, novel small molecules that bind avidly with the surface spike protein of SARS-CoV and thus can interfere with the entry of the virus to its host cells. We achieved this by using a two-step screening method consisting of frontal affinity chromatography-mass spectrometry coupled with a viral infection assay based on a human immunodeficiency virus (HIV)-luc/SARS pseudotyped virus. Two small molecules, tetra-O-galloyl-beta-D-glucose (TGG) and luteolin, were identified, whose anti-SARS-CoV activities were confirmed by using a wild-type SARS-CoV infection system. TGG exhibits prominent anti-SARS-CoV activity with a 50% effective concentration of 4.5 microM and a selective index of 240.0. The two-step screening method described here yielded several small molecules that can be used for developing new classes of anti-SARS-CoV drugs and is potentially useful for the high-throughput screening of drugs inhibiting the entry of HIV, hepatitis C virus, and other insidious viruses into their host cells.  相似文献   
257.
盾叶薯蓣营养器官薯蓣皂甙元含量的动态变化   总被引:2,自引:0,他引:2  
利用高效液相色谱(HPLC)法对盾叶薯蓣营养器官特别是根状茎中薯蓣皂甙元含量的动态变化、品种之间的差异以及雌雄株之间的差异进行了研究。结果表明:实生苗根状茎中薯蓣皂甙元的含量,2年生高于1年生;根茎营养繁殖的2年生根状茎中皂甙元的含量高于1年生的含量;花叶品种的含量高于绿叶品种的含量;雄株的含量比雌株的含量高。在地上的缠绕茎和叶中没有检测到薯蓣皂甙元。由根茎繁殖的1年生根状茎前期皂甙元含量增加缓慢,后期增加较快;2年生根状茎盛花期含量最高,开花后期含量最低,随后含量逐渐增加。为此应在花叶品种中选择产量高、抗性强的品种作为栽培品种。合适的采挖期仍以地上缠绕茎枯萎期为宜。  相似文献   
258.
We measured detailed regional depositionpatterns of inhaled particles in healthy adult male(n = 11; 25 ± 4 yr of age) and female (n = 11; 25 ± 3 yr of age)subjects by means of a serial bolus aerosol delivery technique formonodisperse fine [particle diameter(Dp) = 1 µm] and coarse aerosols(Dp = 3 and 5 µm). The bolus aerosol (40 ml half-width) was delivered to a specificvolumetric depth (Vp) of the lung ranging from 100 to 500 ml with a50-ml increment, and local deposition fraction (LDF) was assessed for each of the 10 local volumetric regions. In all subjects, the deposition distribution pattern was very uneven with respect to Vp,showing characteristic unimodal curves with respect to particle sizeand flow rate. However, the unevenness was more pronounced in women.LDF tended to be greater in all regions of the lung in women than inmen for Dp = 1 µm. For Dp = 3 and 5 µm, LDF showed a marked enhancement in the shallow region of Vp  200 ml in women compared with men(P < 0.05). LDF in women wascomparable to or smaller than those of men in deep lung regions of Vp > 200 ml. Total lung deposition was comparable between men and womenfor fine particles but was consistently greater in women than men forcoarse particles regardless of flow rates used: the difference rangedfrom 9 to 31% and was greater with higher flow rates(P < 0.05). The results indicatethat 1) particledeposition characteristics differ between healthy men and women undercontrolled breathing conditions and2) deposition in women is greaterthan that in men.

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259.
A sensitive and selective high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method was developed for the determination of buagafuran in human plasma. The analyte was extracted from plasma samples with hexane after addition of isotopic internal standard and chromatographed on a RP-C(8) column. The mobile phase consisted of methanol-water (90:10, v/v) and the flow rate was 0.2 mL/min. The detection was performed on a triple quadrupole tandem mass spectrometer in multiple reactions monitoring (MRM) mode using positive electrospray ionization (ESI). The method was validated over the concentration range of 0.5-200 ng/mL. Inter- and intra-day precision (RSD%) were all within 15% and the accuracy (RE%) was equal or lower than 9.5%. The lower limit of quantitation (LLOQ) was 0.5 ng/mL. The extraction recovery was on average 38.1% and the detection was not affected by the matrix. The method was successfully applied to the pharmacokinetic study of buagafuran in healthy Chinese volunteers.  相似文献   
260.
Tang XQ  Fang HR  Li YJ  Zhou CF  Ren YK  Chen RQ  Wang CY  Hu B 《Neurochemical research》2011,36(11):2176-2185
Asymmetric dimethylarginine (ADMA), an endogenous nitric oxide synthase (NOS) inhibitor, is profoundly protective against 1-methy-4-phenylpyridinium ion (MPP+)-induced neurotoxicity. Reactive oxygen species (ROS) overproduction contributes to the neurotoxicity of MPP+; while hydrogen sulfide (H2S) is a pivotal endogenous antioxidant. This study is to assess the potential role of endogenous H2S in the neuroprotection of ADMA against MPP+-induced toxicity in PC12 cells. We showed that ADMA prevented MPP+-induced inhibition of endogenous H2S generation through inhibiting the down-regulation of cystathionine-β-synthetase (CBS, the major enzyme responsible for endogenous H2S generation in PC12 cells) expression and activity elicited by MPP+. ADMA obviously attenuated MPP+-triggered accumulation of intracellular ROS, dissipation of mitochondrial membrane potential (MMP), release of cytochrome c (Cyt-c), and downregulation of Bcl-2 protein expression in PC12 cells. Inhibition of CBS activity by amino-oxyacetate and CBS silencing with a short hairpin RNA vector targeting rat CBS gene reversed the protective action of ADMA against MPP+-caused cytotoxicity, ROS overproduction, and MMP loss in PC12 cells. These results indicate that the protection of ADMA against MPP+-mediated neurotoxicity involves the melioration of MPP+-induced inhibition of endogenous H2S generation. Our findings suggest that modulation of H2S production provide new therapeutic targets for the treatment of neurodegenerative disease, such as Parkinson’s disease.  相似文献   
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