全文获取类型
收费全文 | 4295篇 |
免费 | 300篇 |
出版年
2021年 | 33篇 |
2020年 | 20篇 |
2019年 | 39篇 |
2018年 | 50篇 |
2017年 | 43篇 |
2016年 | 79篇 |
2015年 | 105篇 |
2014年 | 114篇 |
2013年 | 284篇 |
2012年 | 197篇 |
2011年 | 235篇 |
2010年 | 127篇 |
2009年 | 149篇 |
2008年 | 185篇 |
2007年 | 174篇 |
2006年 | 179篇 |
2005年 | 152篇 |
2004年 | 208篇 |
2003年 | 186篇 |
2002年 | 186篇 |
2001年 | 118篇 |
2000年 | 129篇 |
1999年 | 111篇 |
1998年 | 61篇 |
1997年 | 47篇 |
1996年 | 50篇 |
1995年 | 48篇 |
1994年 | 38篇 |
1993年 | 46篇 |
1992年 | 101篇 |
1991年 | 102篇 |
1990年 | 85篇 |
1989年 | 85篇 |
1988年 | 85篇 |
1987年 | 59篇 |
1986年 | 50篇 |
1985年 | 51篇 |
1984年 | 53篇 |
1983年 | 42篇 |
1982年 | 44篇 |
1981年 | 40篇 |
1980年 | 28篇 |
1979年 | 61篇 |
1978年 | 33篇 |
1977年 | 37篇 |
1976年 | 23篇 |
1975年 | 31篇 |
1974年 | 35篇 |
1973年 | 21篇 |
1972年 | 26篇 |
排序方式: 共有4595条查询结果,搜索用时 15 毫秒
91.
Kengo Sasaki Jun Inoue Daisuke Sasaki Namiko Hoshi Tomokazu Shirai Itsuko Fukuda Takeshi Azuma Akihiko Kondo Ro Osawa 《Biotechnology journal》2019,14(5)
Compositional alteration of the gut microbiota is associated with ulcerative colitis (UC). Here, a model culture system is established for the in vitro human colonic microbiota of UC, which will be helpful for determining medical interventions. 16S ribosomal RNA sequencing confirms that UC models are successfully developed from fecal inoculum and retain the bacterial species biodiversity of UC feces. The UC models closely reproduce the microbial components and successfully preserve distinct clusters from the healthy subjects (HS), as observed in the feces. The relative abundance of bacteria belonging to the family Lachnospiraceae significantly decreases in the UC models compared to that in HS, as observed in the feces. The system detects significantly lower butyrogenesis in the UC models than that in HS, correlating with the decreased abundance of Lachnospiraceae. Interestingly, the relative abundance of Lachnospiraceae does not correlate with disease activity (defined as partial Mayo score), suggesting that Lachnospiraceae persists in UC patients at a decreased level, irrespective of the alteration in disease activity. Moreover, the system shows that administration of Clostridium butyricum MIYAIRI restores butyrogenesis in the UC model. Hence, the model detects deregulation in the intestinal environment in UC patients and may be useful for simulating the effect of probiotics. 相似文献
92.
Gorawit Yusakul Seiichi Sakamoto Hiroyuki Tanaka Satoshi Morimoto 《Biotechnology progress》2019,35(4):e2822
The assembly between heavy and light chains is a critical step of immunoglobulin (Ig) and fragment antigen-binding (Fab) antibody expression and of their binding activity. The genes encoding Fab were obtained from hybridoma cells secreting monoclonal antibody (MAb, IgG2b) against adenylate cyclase activator forskolin (FOR). The subclass of the first constant domain of heavy chain (CH1) of IgG2b was modified to IgG1 via overlap extension polymerase chain reaction and expressed via Escherichia coli bacterial system. Since both Fabs (IgG2b and IgG1) were expressed as inclusion bodies, functional analysis was performed after in vitro refolding via stepwise dialysis. The result indicated that the folding efficiency between VH-CH1 and VL-CL was improved by the CH1 modification from IgG2b to IgG1 subclass, although their specificity for FOR was not altered. Effective folding of IgG1 was also observed when they were expressed in the hemolymph of silkworm larvae using the Bombyx mori nuclear polyhedrosis virus bacmid system. An indirect competitive enzyme-linked immunosorbent assay (icELISA) was then developed for the determination of FOR using effectively prepared Fab IgG1. The sensitivity of FOR determination was in the range of 3.91–62.5 ng/mL with less than 9% relative standard deviation, implying the sensitive and reliable analysis of developed icELISA. In addition, high accuracy of the icELISA was supported by the results of spiked-and-recovery tests, ranging from 100.2 to 102.3%. Therefore, Fab could be utilized reliably for icELISA instead of the more expensive MAb. Collectively, this approach improved productivity of Fab and reduced the cost of antibody production. 相似文献
93.
Tanida Kotomi Shimada Mihoko Khor Seik-Soon Toyoda Hiromi Kato Kayoko Kotorii Nozomu Kotorii Tatayu Ariyoshi Yu Kato Takao Hiejima Hiroshi Ozone Motohiro Uchimura Naohisa Ikegami Azusa Kume Kazuhiko Kanbayashi Takashi Imanishi Aya Kamei Yuichi Hida Akiko Wada Yamato Kuroda Kenji Miyamoto Masayuki Hirata Koichi Takami Masanori Yamada Naoto Okawa Masako Omata Naoto Kondo Hideaki Kodama Tohru Inoue Yuichi Mishima Kazuo Honda Makoto Tokunaga Katsushi Miyagawa Taku 《Sleep and biological rhythms》2022,20(1):137-148
Sleep and Biological Rhythms - Idiopathic hypersomnia (IH) is a rare sleep disorder characterized by excessive daytime sleepiness, great difficulty upon awakening, and prolonged sleep time. In... 相似文献
94.
Nitta Masato Kondo Yusuke Ohtsuka Susumu Kamarudin Ahmad Syazni Ismail Norshida 《Systematic parasitology》2022,99(5):587-599
Systematic Parasitology - A new monogenean species, Kannaphallus leptosomus n. sp., from the gills of the diamond trevally, Scyris indica Rüppell, caught off Terengganu, Peninsular Malaysia is... 相似文献
95.
Naoyuki Kondo Mariana Marin Jeong Hwa Kim Tanay M. Desai Gregory B. Melikyan 《The Journal of biological chemistry》2015,290(10):6558-6573
Whether HIV-1 enters cells by fusing with the plasma membrane or with endosomes is a subject of active debate. The ability of HIV-1 to mediate fusion between adjacent cells, a process referred to as “fusion-from-without” (FFWO), shows that this virus can fuse with the plasma membrane. To compare FFWO occurring at the cell surface with HIV-cell fusion through a conventional entry route, we designed an experimental approach that enabled the measurements of both processes in the same sample. The following key differences were observed. First, a very small fraction of viruses fusing with target cells participated in FFWO. Second, whereas HIV-1 fusion with adherent cells was insensitive to actin inhibitors, post-CD4/coreceptor binding steps during FFWO were abrogated. A partial dependence of HIV-cell fusion on actin remodeling was observed in CD4+ T cells, but this effect appeared to be due to the actin dependence of virus uptake. Third, deletion of the cytoplasmic tail of HIV-1 gp41 dramatically enhanced the ability of the virus to promote FFWO, while having a modest effect on virus-cell fusion. Distinct efficiencies and actin dependences of FFWO versus HIV-cell fusion are consistent with the notion that, except for a minor fraction of particles that mediate fusion between the plasma membranes of adjacent cells, HIV-1 enters through an endocytic pathway. We surmise, however, that cell-cell contacts enabling HIV-1 fusion with the plasma membrane could be favored at the sites of high density of target cells, such as lymph nodes. 相似文献
96.
Kai Song Brett H. Herzog Jianxin Fu Minjia Sheng Kirk Bergstrom J. Michael McDaniel Yuji Kondo Samuel McGee Xiaofeng Cai Ping Li Hong Chen Lijun Xia 《The Journal of biological chemistry》2015,290(33):20159-20166
Mucin-type core 1-derived O-glycans, one of the major types of O-glycans, are highly expressed in mammary gland epithelium. Abnormal O-glycans such as Tn antigen are found in over 90% of breast cancers; however, the in vivo role of these aberrant O-glycans in the etiology of breast cancer is unclear. We generated mice with mammary epithelial specific deletion of core 1-derived O-glycans. By crossing with two spontaneous mouse breast cancer models, we determined that loss of core 1-derived O-glycans delays the onset and progression of breast cancer development. Deficiency of core 1 O-glycosylation impaired the localization of Muc1, a major O-glycoprotein, on the apical surfaces of mammary epithelium. Signaling mediated by Muc1, which is critical for breast cancer development, was also defective in the absence of core 1 O-glycans. This study reveals an unexpected role of core 1-derived O-glycans in breast cancer development in mice. 相似文献
97.
98.
99.
Won-Kyung Hong Sun-Yeon Heo Baek-Rock Oh Chul Ho Kim Jung-Hoon Sohn Ji-Won Yang Akihiko Kondo Jeong-Woo Seo 《Bioprocess and biosystems engineering》2013,36(9):1191-1197
In the present study, we established a genetic system for manipulating the oleaginous heterotrophic microalgae Aurantiochytrium sp. KRS101, using cycloheximide resistance as the selectable marker. The gene encoding ribosomal protein L44 (RPL44) of Aurantiochytrium sp. KRS101 was first identified and characterized. Proline 56 was replaced with glutamine, affording cycloheximide resistance to strains encoding the mutant protein. This resistance served as a novel selection marker. The gene encoding the Δ12-fatty acid desaturase of Mortierella alpina, used as a reporter, was successfully introduced into chromosomal DNA of Aurantiochytrium sp. KRS101 via 18S rDNA-targeted homologous recombination. Enzymatic conversion of oleic acid (C18:1) to linoleic acid (C18:2) was detected in transformants but not in the wild-type strain. 相似文献
100.
Y Takubo M Okuda I Takemura F Haruna A Sawatari T Nishihara M Kondo 《Microbiology and immunology》1989,33(7):527-538
It was proved that three spore coat proteins of 48, 36, and 22 kDa (P48, P36, and P22) were the components of the outermost layer (OL) of Bacillus megaterium ATCC 12872 spore by analysis of the isolated OL. And it was indicated that these proteins were deposited not by disulfide bond, but by ionic and/or hydrophobic bonds on the spore. Among them, P36 and P22 were expected to be located on the very surface of the spore by immunological analysis. In the OL deficient mutant of B. megaterium ATCC 12872, MAE05, whose spore was lacking in these OL proteins and galactosamine-6-phosphate polymer, both P36 and P22 were present in the mother cell cytoplasm and deposited on the forespores, but they disappeared with the lysis of mother cells. An OL protein-releasing factor having proteolytic activity was detected in the culture supernatant at the late sporulating stage of both the wild-type and the mutant strains. But the factor could not act on the proteins of the mature spores and the forespores at t10 (tn indicates n hr after the end of exponential growth) of the wild-type strain. Moreover, P36 and P22 were found in the spores of a revertant of MAE05 which could form galactosamine-6-phosphate polymer, suggesting that this sugar polymer played the role in protecting the OL proteins against the protease-like substance after the deposition. 相似文献