全文获取类型
收费全文 | 3955篇 |
免费 | 258篇 |
出版年
2023年 | 5篇 |
2022年 | 25篇 |
2021年 | 43篇 |
2020年 | 26篇 |
2019年 | 41篇 |
2018年 | 59篇 |
2017年 | 55篇 |
2016年 | 84篇 |
2015年 | 128篇 |
2014年 | 168篇 |
2013年 | 219篇 |
2012年 | 247篇 |
2011年 | 254篇 |
2010年 | 184篇 |
2009年 | 172篇 |
2008年 | 276篇 |
2007年 | 251篇 |
2006年 | 254篇 |
2005年 | 248篇 |
2004年 | 252篇 |
2003年 | 195篇 |
2002年 | 230篇 |
2001年 | 71篇 |
2000年 | 81篇 |
1999年 | 76篇 |
1998年 | 50篇 |
1997年 | 47篇 |
1996年 | 34篇 |
1995年 | 21篇 |
1994年 | 28篇 |
1993年 | 25篇 |
1992年 | 35篇 |
1991年 | 31篇 |
1990年 | 32篇 |
1989年 | 31篇 |
1988年 | 23篇 |
1987年 | 22篇 |
1986年 | 29篇 |
1985年 | 26篇 |
1984年 | 23篇 |
1983年 | 17篇 |
1982年 | 14篇 |
1981年 | 7篇 |
1980年 | 8篇 |
1979年 | 16篇 |
1978年 | 11篇 |
1977年 | 15篇 |
1976年 | 4篇 |
1974年 | 3篇 |
1972年 | 5篇 |
排序方式: 共有4213条查询结果,搜索用时 15 毫秒
71.
Olivo Miotto Makoto Sekihara Shin-Ichiro Tachibana Masato Yamauchi Richard D. Pearson Roberto Amato Sonia Gonalves Somya Mehra Rintis Noviyanti Jutta Marfurt Sarah Auburn Ric N. Price Ivo Mueller Mie Ikeda Toshiyuki Mori Makoto Hirai Livingstone Tavul Manuel W. Hetzel Moses Laman Alyssa E. Barry Pascal Ringwald Jun Ohashi Francis Hombhanje Dominic P. Kwiatkowski Toshihiro Mita 《PLoS pathogens》2020,16(12)
The rapid and aggressive spread of artemisinin-resistant Plasmodium falciparum carrying the C580Y mutation in the kelch13 gene is a growing threat to malaria elimination in Southeast Asia, but there is no evidence of their spread to other regions. We conducted cross-sectional surveys in 2016 and 2017 at two clinics in Wewak, Papua New Guinea (PNG) where we identified three infections caused by C580Y mutants among 239 genotyped clinical samples. One of these mutants exhibited the highest survival rate (6.8%) among all parasites surveyed in ring-stage survival assays (RSA) for artemisinin. Analyses of kelch13 flanking regions, and comparisons of deep sequencing data from 389 clinical samples from PNG, Indonesian Papua and Western Cambodia, suggested an independent origin of the Wewak C580Y mutation, showing that the mutants possess several distinctive genetic features. Identity by descent (IBD) showed that multiple portions of the mutants’ genomes share a common origin with parasites found in Indonesian Papua, comprising several mutations within genes previously associated with drug resistance, such as mdr1, ferredoxin, atg18 and pnp. These findings suggest that a P. falciparum lineage circulating on the island of New Guinea has gradually acquired a complex ensemble of variants, including kelch13 C580Y, which have affected the parasites’ drug sensitivity. This worrying development reinforces the need for increased surveillance of the evolving parasite populations on the island, to contain the spread of resistance. 相似文献
72.
Shoko Nakano Yuko Abe Kimiko Nakajima Shigetoshi Sano Osamu Yamamoto Kazumasa Wakamatsu Shosuke Ito Masahiro Hayashi Tamio Suzuki 《Pigment cell & melanoma research》2021,34(1):101-110
Post‐inflammatory hyperpigmentation (PIH) is a common cutaneous condition that can cause a disfigured appearance. However, the pathophysiology of PIH remains poorly understood, at least in part, because an appropriate animal model for research has not been established. In order to analyze the pathomechanism of PIH, we successfully induced PIH in a hairless version of transgenic mice (hk14‐SCF Tg/HRM) that have a human‐type epidermis containing melanin by repeated hapten application of 2,4‐dinitrofluorobenzene. Histopathologic observation showed epidermal hyperplasia, predominant infiltrations of inflammatory cells, and melanin‐containing cells in the dermis just after elicitation of the atopic dermatitis‐like condition. At week 2, the findings were similar to the characteristics of PIH, that is, an increase of melanin without spongiosis or liquid degeneration in the epidermis and an increase in dermal melanophages. Dynamic analysis of melanin showed that the melanin in the dermis remained for a longer duration than in the epidermis. Furthermore, immunohistochemical staining revealed that the majority of cells containing melanin were positive for the anti‐CD68 antibody, but negative for the anti‐F4/80 antibody. These data suggest that novel treatments of PIH should be targeted against macrophages and should eventually lead to the development of new treatment modalities. 相似文献
73.
Epstein-Barr virus (EBV) infection is associated with the development of specific types of lymphoma and some epithelial cancers. EBV infection of resting B-lymphocytes in vitro drives them to proliferate as lymphoblastoid cell lines (LCLs) and serves as a model for studying EBV lymphomagenesis. EBV nuclear antigen 3C (EBNA3C) is one of the genes required for LCL growth and previous work has suggested that suppression of the CDKN2A encoded tumor suppressor p16INK4A and possibly p14ARF is central to EBNA3C’s role in this growth transformation. To directly assess whether loss of p16 and/or p14 was sufficient to explain EBNA3C growth effects, we used CRISPR/Cas9 to disrupt specific CDKN2A exons in EBV transformed LCLs. Disruption of p16 specific exon 1α and the p16/p14 shared exon 2 were each sufficient to restore growth in the absence of EBNA3C. Using EBNA3C conditional LCLs knocked out for either exon 1α or 2, we identified EBNA3C induced and repressed genes. By trans-complementing with EBNA3C mutants, we determined specific genes that require EBNA3C interaction with RBPJ or CtBP for their regulation. Unexpectedly, interaction with the CtBP repressor was required not only for repression, but also for EBNA3C induction of many host genes. Contrary to previously proposed models, we found that EBNA3C does not recruit CtBP to the promoters of these genes. Instead, our results suggest that CtBP is bound to these promoters in the absence of EBNA3C and that EBNA3C interaction with CtBP interferes with the repressive function of CtBP, leading to EBNA3C mediated upregulation. 相似文献
74.
Paulo S. Monzani Samuel Guemra Paulo R. Adona Otavio M. Ohashi Flávio V. Meirelles 《Animal biotechnology》2013,24(2):136-142
Prior to generating transgenic animals for bioreactors, it is important to evaluate the vector constructed to avoid poor protein expression. Mammary epithelial cells cultured in vitro have been proposed as a model to reproduce the biology of the mammary gland. In the present work, three lentiviral vectors were constructed for the human growth hormone (GH), interleukin 2 (IL2), and granulocyte colony-stimulating factor 3 (CSF3) genes driven by the bovine β-casein promoter. The lentiviruses were used to transduce mammary epithelial cells (MAC-T), and the transformed cells were cultured on polystyrene in culture medium with and without prolactin. The gene expression of transgenes was evaluated by PCR using cDNA, and recombinant protein expression was evaluated by Western-blotting using concentrated medium and cellular extracts. The gene expression, of the three introduced genes, was detected in both induced and non induced MAC-T cells. The human GH protein was detected in the concentrated medium, whereas CSF3 was detected in the cellular extract. Apparently, the cellular extract is more appropriate than the concentrated medium to detect recombinant protein, principally because concentrated medium has a high concentration of bovine serum albumin. The results suggest that MAC-T cells may be a good system to evaluate vector construction targeting recombinant protein expression in milk. 相似文献
75.
Hiroaki Horikawa Midori Okada Yuko Nakamura Azusa Sato Noriko Iwamoto 《Free radical research》2013,47(10):1059-1065
The chemical properties of Amadori compounds in the presence of transition metal ions were studied, using the analogs 1-deoxy-1- n -butylamino- d -fructose (DBF) and N f -formyl-fructoselysine (fFL). The following characteristics were revealed: (a) DBF combined easily with Cu 2+ (but no other transition metal ions) to form a DBF-Cu 2+ complex in phosphate buffer, pH 7.4; (b) the complex was unstable, and degraded with the release of Cu + during incubation at 37°C; (c) degradation of the complex was associated with the production of hydroxyl radicals by the Fenton reaction and f -dicarbonyl compounds by non-autoxidative degradation; and (d) properties of DBF were similar to those of fFL. The above properties were additionally observed in glycated poly-Lys (GPL). Our findings indicate a novel mechanism for the generation of hydroxyl radicals and f -dicarbonyl compounds from Amadori adducts in the presence of Cu 2+ . 相似文献
76.
Nobuo N Noda Yuko Fujioka Takao Hanada Yoshinori Ohsumi Fuyuhiko Inagaki 《EMBO reports》2013,14(2):206-211
Atg12 is conjugated to Atg5 through enzymatic reactions similar to ubiquitination. The Atg12–Atg5 conjugate functions as an E3‐like enzyme to promote lipidation of Atg8, whereas lipidated Atg8 has essential roles in both autophagosome formation and selective cargo recognition during autophagy. However, the molecular role of Atg12 modification in these processes has remained elusive. Here, we report the crystal structure of the Atg12–Atg5 conjugate. In addition to the isopeptide linkage, Atg12 forms hydrophobic and hydrophilic interactions with Atg5, thereby fixing its position on Atg5. Structural comparison with unmodified Atg5 and mutational analyses showed that Atg12 modification neither induces a conformational change in Atg5 nor creates a functionally important architecture. Rather, Atg12 functions as a binding module for Atg3, the E2 enzyme for Atg8, thus endowing Atg5 with the ability to interact with Atg3 to facilitate Atg8 lipidation. 相似文献
77.
Sachiko N. Isobe Hideki Hirakawa Shusei Sato Fumi Maeda Masami Ishikawa Toshiki Mori Yuko Yamamoto Kenta Shirasawa Mitsuhiro Kimura Masanobu Fukami Fujio Hashizume Tomoko Tsuji Shigemi Sasamoto Midori Kato Keiko Nanri Hisano Tsuruoka Chiharu Minami Chika Takahashi Tsuyuko Wada Akiko Ono Kumiko Kawashima Naomi Nakazaki Yoshie Kishida Mitsuyo Kohara Shinobu Nakayama Manabu Yamada Tsunakazu Fujishiro Akiko Watanabe Satoshi Tabata 《DNA research》2013,20(1):79-92
78.
Yuko Kimura Noboru Takamura Masato Fukunaga Mitsuo Kanagae Yasuyo Abe Kiyoshi Aoyagi 《Biomarkers》2013,18(3):291-297
Primary objective: To carry out a preliminary evaluation of subclinical inflammation and its genetic background in young adults.Research design: Fifty-five healthy Japanese young adults aged 19–27 years (37 males and 18 females, mean age: 22.3 years), and 58 healthy Japanese adults aged 40 to 60 years (21 males and 37 females, mean age: 51.5 years) were included in this study.Methods and procedures: We measured plasma high-sensitive C-reactive protein (hs-CRP) levels and screened for the C677T polymorphism of the 5-10 methylenetetrahydrofolate reductase gene (MTHFR), which is considered a genetic risk factor for atherosclerosis, by HinfI digestion.Main outcomes and results: Hs-CRP levels of the young adult group were significantly lower than the levels of the middle aged group (0.014±0.030 mg/dl vs. 0.031±0.040 mg/dl, p=0.005). The levels were significantly higher in males than in females (0.028±0.019 mg/dl vs. 0.013±0.010 mg/dl, p=0.008) among young adults. Furthermore, we evaluated the relationship of the C677T genotype and hs-CRP values, but found no association between them.Conclusions: Although the sample size is limited, our preliminary study demonstrated the profiles of hs-CRP in Japanese young adults. Further investigation will be needed to establish the guidelines for customized school health education using sensitive laboratory and genetic markers. 相似文献
79.
Toru Sugita Yukari Semi Hiromasa Sawada Yumi Utoyama Yuko Hosomi Eri Yoshimoto Yuki Maehata Hiroyuki Fukuoka Ryutaro Nagata Akio Ohyama 《Molecular breeding : new strategies in plant improvement》2013,31(4):909-920
To facilitate marker-assisted breeding and genetic analyses of pepper (Capsicum annuum), we developed non-redundant 2- or 3-base simple sequence repeat (SSR) markers from enriched C. annuum genomic libraries and from C. annuum cDNA sequences in public databases. The SSR-enriched libraries were constructed using combinations of three restriction enzymes (AluI, HaeIII, and RsaI) and two biotinylated oligonucleotides [b(GA)15 and b(CA)15]. Ultimately, we obtained 1,736 genomic SSR markers and 1,344 cDNA-derived SSR markers from 6,528 clones and 13,003 sequences, respectively. We mapped 597 markers, including 265 of the newly developed SSR markers, onto a linkage map by using doubled-haploid (DH) lines derived from an intraspecific cross of two pure lines of C. annuum (K9-11 × MZC-180). The map, designated as the KL-DH map, consisted of 12 linkage groups. The map covered a genetic distance of 2,028 cM, and the average distance between markers was less than 4 cM. The frame structure of the KL-DH map was compared with the published standard conserved ortholog set II (COSII) map, which was derived from an interspecific F2 population (C. frutescens × C. annuum), by using tomato (Solanum lycopersicum) chromosomal sequences to bridge the two maps. The intraspecific KL-DH map constructed in this study and the interspecific COSII map were similar in map length and marker distribution, suggesting that the KL-DH map covers nearly the whole genome of C. annuum. 相似文献