全文获取类型
收费全文 | 1153篇 |
免费 | 60篇 |
国内免费 | 68篇 |
出版年
2023年 | 18篇 |
2022年 | 13篇 |
2021年 | 23篇 |
2020年 | 38篇 |
2019年 | 41篇 |
2018年 | 44篇 |
2017年 | 29篇 |
2016年 | 29篇 |
2015年 | 26篇 |
2014年 | 61篇 |
2013年 | 109篇 |
2012年 | 42篇 |
2011年 | 45篇 |
2010年 | 34篇 |
2009年 | 54篇 |
2008年 | 45篇 |
2007年 | 67篇 |
2006年 | 44篇 |
2005年 | 49篇 |
2004年 | 40篇 |
2003年 | 27篇 |
2002年 | 32篇 |
2001年 | 31篇 |
2000年 | 20篇 |
1999年 | 27篇 |
1998年 | 17篇 |
1997年 | 22篇 |
1996年 | 12篇 |
1995年 | 23篇 |
1994年 | 26篇 |
1993年 | 22篇 |
1992年 | 15篇 |
1991年 | 20篇 |
1990年 | 17篇 |
1989年 | 10篇 |
1988年 | 12篇 |
1987年 | 11篇 |
1986年 | 8篇 |
1985年 | 8篇 |
1984年 | 14篇 |
1983年 | 10篇 |
1982年 | 4篇 |
1981年 | 5篇 |
1980年 | 4篇 |
1979年 | 9篇 |
1978年 | 6篇 |
1977年 | 4篇 |
1976年 | 8篇 |
1975年 | 3篇 |
1973年 | 1篇 |
排序方式: 共有1281条查询结果,搜索用时 953 毫秒
991.
992.
Matthias Brunner Philipp Brosig Monika Losing Marco Kunzelmann Amandine Calvet Fabian Stiefel Jan Bechmann Andreas Unsoeld Jochen Schaub 《Engineering in Life Science》2019,19(10):666-680
Biopharmaceutical manufacturing processes can be affected by variability in cell culture media, e.g. caused by raw material impurities. Although efforts have been made in industry and academia to characterize cell culture media and raw materials with advanced analytics, the process of industrial cell culture media preparation itself has not been reported so far. Within this publication, we first compare mid‐infrared and two‐dimensional fluorescence spectroscopy with respect to their suitability as online monitoring tools during cell culture media preparation, followed by a thorough assessment of the impact of preparation parameters on media quality. Through the application of spectroscopic methods, we can show that media variability and its corresponding root cause can be detected online during the preparation process. This methodology is a powerful tool to avoid batch failure and is a valuable technology for media troubleshooting activities. Moreover, in a design of experiments approach, including additional liquid chromatography–mass spectrometry analytics, it is shown that variable preparation parameters such as temperature, power input and preparation time can have a strong impact on the physico‐chemical composition of the media. The effect on cell culture process performance and product quality in subsequent fed‐batch processes was also investigated. The presented results reveal the need for online spectroscopic methods during the preparation process and show that media variability can already be introduced by variation in media preparation parameters, with a potential impact on scale‐up to a commercial manufacturing process. 相似文献
993.
传统公园评价的研究大多集中在单个公园使用满意度方面,存在问卷样本量小、采集时间较集中等缺陷。大数据相关研究的快速发展为公园系统评价提供了新的契机。基于对社交媒体数据的文本分析,研究对比广州市 50 个公园的评价结果,同时探究公园评价与人口统计学特征、地理区位、评价时间以及公园属性之间的关系。研究发现, 1)公园正面评价主要集中在免费设施、环境和特色等方面,而负面评价多集中于公园卫生、交通成本、停车等管理服务方面,广州公园的管理服务水平亟待提升。 2)女性较男性更关注公园能否满足其对社交场所的需求,本地游客较外地游客更关注公园本身的服务质量而非公园的外部属性。 3)基干公园应更侧重其可达性和基本的服务功能,承担大众日常活动;特色公园应更注重满足特定人群的需求及特色打造;自然公园应更关注公园生态质量并能够承担需要较大场地的活动。 4) 2010—2018 年,人们越来越关注公园特色的打造,在公园中的活动类型也越来越多样,现代公园建设更应着力于文化服务质量的提升。研究突破传统公园研究方法的局限,将大数据的手段运用到多个公园的整体评价当中,有助于建立更普适、更有针对性的公园评价指标体系,具有较高的实践价值。 相似文献
994.
目的:通过报道1例中耳术后并发耳廓软骨膜炎致耳廓畸形患者的病历资料,以加强临床医师对该疾病的认识与防范。方法:对本例患者的病历资料进行回顾性阐述,结合该患者的临床特点揭示中耳术后并发耳廓软骨膜炎的发生发展过程,并通过文献回顾阐述该疾病的发病原因、病原学特点及治疗方法。结果:耳廓软骨膜炎是耳科手术术后较为棘手但相对少见的并发症,最为常见铜绿假单胞杆菌感染所致。治疗方法视患者病情而定,包括抗菌药物应用、脓肿切开引流、病变软骨切除术。对于合并糖尿病的老年患者中耳术后更易引发耳廓软骨膜炎,且疗程更长、预后更差。结论:中耳术后并发耳廓软骨膜炎临床需警惕,对合并糖尿病的老年患者应多加防范;早诊断,早治疗,避免增加患者痛苦、加重其经济负担,并与患者做好充分沟通,避免医疗纠纷的发生。 相似文献
995.
Two‐liquid‐phase reaction media have long been used in bioconversions to supply or remove hydrophobic organic reaction substrates and products to reduce inhibitory and toxic effects on biocatalysts. In case of the terminal oxyfunctionalization of linear alkanes by the AlkBGT monooxygenase the excess alkane substrate is often used as a second phase to extract the alcohol, aldehyde, and acid products. However, the selection of other carrier phases or surfactants is complex due to a large number of parameters that are involved, such as biocompatibility, substrate bioavailability, and product extraction selectivity. This study combines systematic high‐throughput screening with chemometrics to correlate physicochemical parameters of a range of cosolvents to product specificity and yield using a multivariate regression model. Partial least‐squares regression shows that the defining factor for product specificity is the solubility properties of the reaction substrate and product in the cosolvent, as measured by Hansen solubility parameters. Thus the polarity of cosolvents determines the accumulation of either alcohol or acid products. Whereas usually the acid product accumulates during the reaction, by choosing a more polar cosolvent the 1‐alcohol product can be accumulated. Especially with Tergitol as a cosolvent, a 3.2‐fold improvement in the 1‐octanol yield to 18.3 mmol L?1 is achieved relative to the control reaction without cosolvents. 相似文献
996.
Abstract The electrophoretical separations of Mycoplasma arthritidis and the serum used in the cultivation medium show a high number of protein bands with identical molecular weights. Proteins with molecular weights of 84, 72 and 52 kDa also appeared to be identical with proteins of Mycoplasma arthritidis in their antigenic properties as demonstrated by Western blotting with rat-anti- Mycoplasma arthritidis serum. The autoradiography of electrophoretically separated Mycoplasma arthritidis cells metabolically labeled with 35 S-methionine and 35 S-cysteine revealed that the proteins of Mycoplasma arthiritidis identical in molecular weight and antigenic structure with serum proteins are synthesized by Mycoplasma arthritidis , and represent true translation products. 相似文献
997.
Tingzhou Zhang Lirong Yang Ziqiang Zhu Jianping Wu 《Journal of Molecular Catalysis .B, Enzymatic》2002,18(4-6)
Optically active form of α-cyano-3-phenoxybenzyl alcohol (CPBA), building block of pyrethroid insecticides, was synthesized as its acetate by the combination of anion-exchange resin-catalyzed transcyanation between m-phenoxybenzaldehyde and acetone cyanohydrin, and lipase-catalyzed enantioselective transesterification of the resulting cyanohydrin with vinyl acetate. Through a screening of enzymes, Alcaligenes sp. lipase showed the highest activity and the conversion exceeded 50%. Effects of solvents and temperatures on this reaction were studied. Among four kinds of solvent, diisopropyl ether was a best choice. The optimal temperature was 50 °C. Although the external diffusion limitation could be excluded by raising the rotational speed, internal diffusion could not be ignored, since the enzyme was an immobilized one on the particles with a considerably large diameter. The e.e. values of CPBA ester were measured by polarimeter and NMR. 相似文献
998.
Lucid dreams, as well as control dreams, have recently been reported as associated with video game play (Gackenbach, 2006). In this study, dreams were collected from the morning after a night of rested sleep as well as electronic media use from the day before the dream. In a factor analysis, lucid and control dreams were associated with all electronic media use but most strongly with video game play. (PsycINFO Database Record (c) 2010 APA, all rights reserved) 相似文献
999.
Menin plays an established role in the differentiation of mesenchymal cells to the osteogenic lineage. Conversely, whether Menin influences the commitment of mesenschymal cells to the myogenic lineage, despite expression in the developing somite was previously unclear. We observed that Menin is down-regulated in C2C12 and C3H10T1/2 mesenchymal cells when muscle differentiation is induced. Moreover, maintenance of Menin expression by constitutive ectopic expression inhibited muscle cell differentiation. Reduction of Menin expression by siRNA technology results in precocious muscle differentiation and concomitantly attenuates BMP-2 induced osteogenesis. Reduced Menin expression antagonizes BMP-2 and TGF-β1 mediated inhibition of myogenesis. Furthermore, Menin was found to directly interact with and potentiate the transactivation properties of Smad3 in response to TGF-β1. Finally in concert with these observations, tissue-specific inactivation of Men1 in Pax3-expressing somite precursor cells leads to a patterning defect of rib formation and increased muscle mass in the intercostal region. These data invoke a pivotal role for Menin in the competence of mesenchymal cells to respond to TGF-β1 and BMP-2 signals. Thus, by modulating cytokine responsiveness Menin functions to alter the balance of multipotent mesenchymal cell commitment to the osteogenic or myogenic lineages. 相似文献
1000.
Doina Popov Miruna Nemecz Adriana Georgescu 《Biochemical and biophysical research communications》2009,388(1):51-1126
Hyperglycemia stimulates a plethora of intracellular signaling pathways within the cells of the vascular wall resulting in dysfunction-associated pathologies. Most of the studies reported so far explored the effect of rather short-time exposure of smooth muscle cells to high glucose concentrations. To mimic situation in Type 2 diabetes in which vascular wall is constantly exposed to circulating hyperglycemia, we report here the long-term (7 days) effect of high glucose concentration on human media artery smooth muscle cells. This consists in up-regulation of PTP1B protein expression, down-regulation of basal Akt phosphorylation, and elevation of basal ERK1/2 activation. Acute stimulation of cells in high glucose with insulin down-regulated PTP1B expression, slightly decreased ERK1/2 activity, and activated Akt, whereas oxidative stress up-regulated Akt and ERK1/2 phosphorylation. In conclusion, long-term high glucose and acute oxidative stress and insulin stimulation imbalance the expression of activated kinases Akt and ERK1/2 and of dephosphorylating PTP1B in the insulin signaling pathway. 相似文献