全文获取类型
收费全文 | 2194篇 |
免费 | 126篇 |
出版年
2022年 | 6篇 |
2021年 | 25篇 |
2020年 | 16篇 |
2019年 | 15篇 |
2018年 | 23篇 |
2017年 | 22篇 |
2016年 | 40篇 |
2015年 | 65篇 |
2014年 | 68篇 |
2013年 | 168篇 |
2012年 | 138篇 |
2011年 | 135篇 |
2010年 | 82篇 |
2009年 | 93篇 |
2008年 | 150篇 |
2007年 | 133篇 |
2006年 | 125篇 |
2005年 | 118篇 |
2004年 | 146篇 |
2003年 | 131篇 |
2002年 | 118篇 |
2001年 | 22篇 |
2000年 | 31篇 |
1999年 | 32篇 |
1998年 | 32篇 |
1997年 | 24篇 |
1996年 | 26篇 |
1995年 | 20篇 |
1994年 | 15篇 |
1993年 | 13篇 |
1992年 | 19篇 |
1991年 | 21篇 |
1990年 | 25篇 |
1989年 | 17篇 |
1988年 | 18篇 |
1987年 | 21篇 |
1986年 | 13篇 |
1985年 | 11篇 |
1984年 | 9篇 |
1983年 | 13篇 |
1982年 | 11篇 |
1980年 | 15篇 |
1979年 | 10篇 |
1978年 | 9篇 |
1977年 | 9篇 |
1976年 | 6篇 |
1975年 | 11篇 |
1973年 | 7篇 |
1972年 | 8篇 |
1970年 | 7篇 |
排序方式: 共有2320条查询结果,搜索用时 31 毫秒
81.
Sujin Bao Libin Zhu Qiang Zhuang Lucia Wang Pin-Xian Xu Keiji Itoh Ian R. Holzman Jing Lin 《PloS one》2013,8(3)
One approach to deliver therapeutic agents, especially proteins, to the gastro-intestinal (GI) tract is to use commensal bacteria as a carrier. Genus Lactobacillus is an attractive candidate for use in this approach. However, a system for expressing exogenous proteins at a high level has been lacking in Lactobacillus. Moreover, it will be necessary to introduce the recombinant Lactobacillus into the GI tract, ideally by oral administration. Whether orally administered Lactobacillus can reach and reside in the GI tract has not been explored in neonates. In this study, we have examined these issues in neonatal rats. To achieve a high level of protein expression in Lactobacillus, we tested the impact of three promoters and two backbones on protein expression levels using mRFP1, a red fluorescent protein, as a reporter. We found that a combination of an L-lactate dehydrogenase (ldhL) promoter of Lactobacillus sakei with a backbone from pLEM415 yielded the highest level of reporter expression. When this construct was used to transform Lactobacillus casei, Lactobacillus delbrueckii and Lactobacillus acidophilus, high levels of mRFP1 were detected in all these species and colonies of transformed Lactobacillus appeared pink under visible light. To test whether orally administered Lactobacillus can be retained in the GI tract of neonates, we fed the recombinant Lactobacillus casei to neonatal rats. We found that about 3% of the bacteria were retained in the GI tract of the rats at 24 h after oral feeding with more recombinant Lactobacillus in the stomach and small intestine than in the cecum and colon. No mortality was observed throughout this study with Lactobacillus. In contrast, all neonatal rats died within 24 hours after fed with transformed E. coli. Taken together, our results indicate that Lactobacillus has the potential to be used as a vehicle for the delivery of therapeutic agents to neonates. 相似文献
82.
Yasuo Noda Yoshiki Ida Shinichi Tanaka Taku Toyama Murilo Felix Roggia Yasuhiro Tamaki Naohiko Sugita Mamoru Mitsuishi Takashi Ueta 《PloS one》2013,8(1)
Purpose
To elucidate the merits of robotic application for vitreoretinal maneuver in comparison to conventional manual performance using an in-vitro eye model constructed for the present study.Methods
Capability to accurately approach the target on the fundus, to stabilize the manipulator tip just above the fundus, and to perceive the contact of the manipulator tip with the fundus were tested. The accuracies were compared between the robotic and manual control, as well as between ophthalmologists and engineering students.Results
In case of manual control, ophthalmologists were superior to engineering students in all the 3 test procedures. Robotic assistance significantly improved accuracy of all the test procedures performed by engineering students. For the ophthalmologists including a specialist of vitreoretinal surgery, robotic assistance enhanced the accuracy in the stabilization of manipulator tip (from 90.9 µm to 14.9 µm, P = 0.0006) and the perception of contact with the fundus (from 20.0 mN to 7.84 mN, P = 0.046), while robotic assistance did not improve pointing accuracy.Conclusions
It was confirmed that telerobotic assistance has a potential to significantly improve precision in vitreoretinal procedures in both experienced and inexperienced hands. 相似文献83.
Masatoshi Teraguchi Noriko Yoshimura Hiroshi Hashizume Shigeyuki Muraki Hiroshi Yamada Hiroyuki Oka Akihito Minamide Yuyu Ishimoto Keiji Nagata Ryohei Kagotani Sakae Tanaka Hiroshi Kawaguchi Kozo Nakamura Toru Akune Munehito Yoshida 《PloS one》2016,11(2)
Objective
The objective of the present study was to examine the associations between metabolic syndrome (MS) components, such as overweight (OW), hypertension (HT), dyslipidemia (DL), and impaired glucose tolerance (IGT), and intervertebral disc degeneration (DD).Design
The present study included 928 participants (308 men, 620 women) of the 1,011 participants in the Wakayama Spine Study. DD on magnetic resonance imaging was classified according to the Pfirrmann system. OW, HT, DL, and IGT were assessed using the criteria of the Examination Committee of Criteria for MS in Japan.Results
Multivariable logistic regression analysis revealed that OW was significantly associated with cervical, thoracic, and lumbar DD (cervical: odds ratio [OR], 1.28; 95% confidence interval [CI], 0.92–1.78; thoracic: OR, 1.75; 95% CI, 1.24–2.51; lumbar: OR, 1.87; 95% CI, 1.06–3.48). HT and IGT were significantly associated with thoracic DD (HT: OR, 1.54; 95% CI, 1.09–2.18; IGT: OR, 1.65; 95% CI, 1.12–2.48). Furthermore, subjects with 1 or more MS components had a higher OR for thoracic DD compared with those without MS components (vs. no component; 1 component: OR, 1.58; 95% CI, 1.03–2.42; 2 components: OR, 2.60; 95% CI, 1.62–4.20; ≥3 components: OR, 2.62; 95% CI, 1.42–5.00).Conclusion
MS components were significantly associated with thoracic DD. Furthermore, accumulation of MS components significantly increased the OR for thoracic DD. These findings support the need for further studies of the effects of metabolic abnormality on DD. 相似文献84.
85.
Yasuko Sekita Keiji Murakami Hiromichi Yumoto Hiroyuki Mizuguchi Takashi Amoh Satoshi Ogino 《Bioscience, biotechnology, and biochemistry》2016,80(6):1205-1213
Houttuynia cordata (HC) has been commonly used as many traditional remedies in local areas of Japan. Although many pharmacological activities of HC have been reported, the mechanism underlying the effect of HC remains unknown. We conducted the interview survey in Japan to verify how HC was actually used. The interview survey revealed that HC poultice (HCP) prepared from smothering fresh leaves of HC was most frequently used for the treatment of purulent skin diseases including furuncle and carbuncle with high effectiveness. Ethanol extract of HCP (eHCP) showed anti-bacterial effects against methicillin-resistant Staphylococcus aureus (MRSA), and showed an anti-biofilm activity against MRSA. eHCP showed dose-dependent inhibition of S. aureus lipoteichoic acid (LTA)-induced interleukin-8 and CCL20 production in human keratinocyte without any cytotoxicity. These results suggest that HCP is effective for skin abscess and its underlying mechanism might be the complicated multiple activities for both bacteria and host cells. 相似文献
86.
Kimata-Ariga Y Kurisu G Kusunoki M Aoki S Sato D Kobayashi T Kita K Horii T Hase T 《Journal of biochemistry》2007,141(3):421-428
The human malaria parasite (Plasmodium falciparum) possesses a plastid-derived organelle called the apicoplast, which is believed to employ metabolisms crucial for the parasite's survival. We cloned and studied the biochemical properties of plant-type ferredoxin (Fd) and Fd-NADP+ reductase (FNR), a redox system that potentially supplies reducing power to Fd-dependent metabolic pathways in malaria parasite apicoplasts. The recombinant P. falciparum Fd and FNR proteins were produced by synthetic genes with altered codon usages preferred in Escherichia coli. The redox potential of the Fd was shown to be considerably more positive than those of leaf-type and root-type Fds from plants, which is favourable for a presumed direction of electron flow from catabolically generated NADPH to Fd in the apicoplast. The backbone structure of P. falciparum Fd, as solved by X-ray crystallography, closely resembles those of Fds from plants, and the surface-charge distribution shows several acidic regions in common with plant Fds and some basic regions unique to this Fd. P. falciparum FNR was able to transfer electrons selectively to P. falciparum Fd in a reconstituted system of NADPH-dependent cytochrome c reduction. These results indicate that an NADPH-FNR-Fd cascade is operative in the apicoplast of human malaria parasites. 相似文献
87.
Fbs1 is an F-box protein present abundantly in the nervous system. Similar to the ubiquitously expressed Fbs2, Fbs1 recognizes N-glycans at the innermost position as a signal for unfolded glycoproteins, probably in the endoplasmic reticulum-associated degradation pathway. Here, we show that the in vivo majority of Fbs1 is present as Fbs1-Skp1 heterodimers or Fbs1 monomers but not SCF(Fbs1) complex. The inefficient SCF complex formation of Fbs1 and the restricted presence of SCF(Fbs1) bound on the endoplasmic reticulum membrane were due to the short linker sequence between the F-box domain and the sugar-binding domain. In vitro, Fbs1 prevented the aggregation of the glycoprotein through the N-terminal unique sequence of Fbs1. Our results suggest that Fbs1 assists clearance of aberrant glycoproteins in neuronal cells by suppressing aggregates formation, independent of ubiquitin ligase activity, and thus functions as a unique chaperone for those proteins. 相似文献
88.
Background
This paper reports on the modification of the Parental Nurturance Scale (PNS), translation of the modified version (PNSM) from English to Japanese, and equivalence assessment between the PNSM and the translated version (PNSM-J). The PNS was modified so as to enable its use in nurturance studies where the prime source of nurturance might vary between respondents. 相似文献89.
90.