全文获取类型
收费全文 | 1880篇 |
免费 | 88篇 |
国内免费 | 4篇 |
专业分类
林业 | 203篇 |
农学 | 53篇 |
基础科学 | 4篇 |
280篇 | |
综合类 | 76篇 |
农作物 | 88篇 |
水产渔业 | 230篇 |
畜牧兽医 | 902篇 |
园艺 | 17篇 |
植物保护 | 119篇 |
出版年
2023年 | 1篇 |
2022年 | 17篇 |
2021年 | 47篇 |
2020年 | 16篇 |
2019年 | 32篇 |
2018年 | 39篇 |
2017年 | 49篇 |
2016年 | 62篇 |
2015年 | 76篇 |
2014年 | 91篇 |
2013年 | 141篇 |
2012年 | 102篇 |
2011年 | 138篇 |
2010年 | 59篇 |
2009年 | 70篇 |
2008年 | 121篇 |
2007年 | 122篇 |
2006年 | 129篇 |
2005年 | 119篇 |
2004年 | 121篇 |
2003年 | 117篇 |
2002年 | 84篇 |
2001年 | 67篇 |
2000年 | 41篇 |
1999年 | 25篇 |
1998年 | 13篇 |
1997年 | 11篇 |
1996年 | 3篇 |
1995年 | 7篇 |
1994年 | 5篇 |
1993年 | 4篇 |
1992年 | 2篇 |
1991年 | 6篇 |
1990年 | 4篇 |
1989年 | 4篇 |
1988年 | 1篇 |
1987年 | 4篇 |
1986年 | 4篇 |
1985年 | 1篇 |
1984年 | 3篇 |
1983年 | 2篇 |
1982年 | 1篇 |
1981年 | 1篇 |
1979年 | 3篇 |
1977年 | 1篇 |
1974年 | 2篇 |
1973年 | 1篇 |
1972年 | 1篇 |
1971年 | 1篇 |
1970年 | 1篇 |
排序方式: 共有1972条查询结果,搜索用时 31 毫秒
21.
Tomioka I Mizutani E Yoshida T Sugawara A Inai K Sasada H Sato E 《The Journal of reproduction and development》2007,53(4):835-842
The present study was conducted to demonstrate the spindle formation and behavior of chromosomes and microtubules during first division in reconstructed rat embryos produced by somatic cell nuclear transfer (SCNT) with cumulus cell nuclei. To demonstrate the effect of oocyte aging after ovulation on the cleavage of SCNT embryos, micromanipulation was carried out 11, 15 and 18 h after injection of hCG. SCNT oocytes were activated by incubation in culture medium supplemented with 5 microM ionomycin for 5 min followed by treatment with 2 mM 6-dimethylaminopurine (6-DMAP) in mR1ECM for 2-3 h. For immunocytochemical observation, the SCNT embryos were incubated with monoclonal anti-alpha-tubulin antibody and then fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse IgG. Cleavage rates were significantly higher for oocytes collected after 15 and 18 h rather than for those collected 11 h after injection of hCG (56 and 53%, respectively vs. 28%; P<0.05). Premature chromosome condensation occurred before activation of the SCNT oocytes, but adequate spindle formation was only rarely observed. The distribution of microtubules in SCNT embryos after activation was different from those of fertilized and parthenogenic oocytes, i.e., a dense microtubule organization shaped like a ring was observed. Eighteen to 20 h post-activation, most SCNT embryos were in the 2-cell stage, but no nucleoli were clearly visible, which was quite different from the fertilized oocytes. In addition, first division with and without small cellular bodies containing DNA was observed in the rat SCNT embryos in some cases. The present study suggests that reorganization of transferred nuclei in rat SCNT embryos may be inadequate in terms of formation of the mitotic assembly and nucleolar reorganization. 相似文献
22.
23.
Kobayashi Y Inoue N Sato G Itou T Santos HP Brito CJ Gomes AA Santos MF Silva MV Mota CS Ito FH Sakai T 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2007,69(7):691-696
The incidence of canine rabies has been widely reported in Brazil, and new rabies virus (RV) variants, genetically similar to canine RV, have recently been isolated from foxes. In order to derive the epidemiological characteristics of Brazilian Carnivora RV, Brazilian RVs isolated from dogs, cats, and foxes were genetically analyzed. Brazilian Carnivora RV isolates were divided into 2 main lineages. The predominant lineage was found in dogs and cats, which included the Argentinean and Bolivian Carnivora RV isolates, and was extensively distributed throughout Brazil and surrounding countries. The other lineage consisted of three sublineages containing Brazilian dog and fox RV isolates, with the dog sublineages located on an internal branch of 2 fox sublineages, suggesting that RV transmission events might have occurred between foxes and dogs in the past. These results suggest that contact between dogs and wildlife has the potential to generate new rabies variants and that it is important to control RV infection cycles in both dogs and wildlife to prevent spread of rabies infection. 相似文献
24.
A novel culture method of canine peripheral blood lymphocytes with concanavalin a and recombinant human interleukin-2 for adoptive immunotherapy 总被引:1,自引:0,他引:1
Kato M Watarai S Nishikawa S Iwasaki T Kodama H 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2007,69(5):481-486
This study was designed to develop a novel culture method for the efficient proliferation of canine peripheral blood lymphocytes (cPBL) for adoptive immunotherapy. When cPBL were cultured in the presence of concanavalin A (Con A), proliferation of cPBL was induced and expression of interleukin-2 receptor (IL-2R) which enables to respond to exogenously added IL-2 was upregulated. And then, when cPBL were cultured with recombinant human interleukin-2 (rhIL-2) in addition to Con A, proliferation was accelerated and increased to about 10-fold after 1 week. The phenotypic analysis showed that the main population of the cultured cPBL was consisted of CD8+ positive lymphocytes. Among them, CD4+CD8+ double positive (DP) lymphocytes had significantly increased, and the ratio of CD4+ single positive (SP) lymphocytes to CD8+ SP lymphocytes (CD4+SP/CD8+SP) was decreased as compared to before culturing. To evaluate the cytotoxic activity of cPBL cultured with Con A and rhIL-2, furthermore, cytotoxic assay was carried out against xenogeneic melanoma cell line (MeWo), which resulted in MHC-unrestricted cytokilling. These results suggest that the culture method of cPBL by the use of Con A and rhIL-2 may be useful for generating lymphokine activated killer cells, and also this may be beneficial for adoptive immunotherapy of tumor-bearing dogs. 相似文献
25.
Kosuke SODA Maya YAMANE Chiharu HIDAKA Kozue MIURA Trang T. H. UNG Hang L. K. NGUYEN Hiroshi ITO Mai Q. LE Toshihiro ITO 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2021,83(12):1899
Low and highly pathogenic avian influenza viruses (LPAIVs and HPAIVs, respectively) have been co-circulating in poultry populations in Asian, Middle Eastern, and African countries. In our avian-flu surveillance in Vietnamese domestic ducks, viral genes of LPAIV and HPAIV have been frequently detected in the same individual. To assess the influence of LPAIV on the pathogenicity of H5 HPAIV in domestic ducks, an experimental co-infection study was performed. One-week-old domestic ducks were inoculated intranasally and orally with phosphate-buffered saline (PBS) (control) or 106 EID50 of LPAIVs (A/duck/Vietnam/LBM678/2014 (H6N6) or A/Muscovy duck/Vietnam/LBM694/2014 (H9N2)). Seven days later, these ducks were inoculated with HPAIV (A/Muscovy duck/Vietnam/LBM808/2015 (H5N6)) in the same manner. The respective survival rates were 100% and 50% in ducks pre-infected with LBM694 or LBM678 strains and both higher than the survival of the control group (25%). The virus titers in oral/cloacal swabs of each LPAIV pre-inoculation group were significantly lower at 3–5 days post-HPAIV inoculation. Notably, almost no virus was detected in swabs from surviving individuals of the LBM678 pre-inoculation group. Antigenic cross-reactivity among the viruses was not observed in the neutralization test. These results suggest that pre-infection with LPAIV attenuates the pathogenicity of HPAIV in domestic ducks, which might be explained by innate and/or cell-mediated immunity induced by the initial infection with LPAIV. 相似文献
26.
Chiharu HIDAKA Kosuke SODA Toshihiro ITO Hiroshi ITO 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2022,84(1):121
Although verogenic Newcastle disease viruses (NDVs) generally cause subclinical infection in waterfowls such as ducks, NDVs with high virulence in waterfowl have been sporadically reported. We previously reported that the NDV d5a20b strain, which is obtained by serial passaging of the velogenic 9a5b strain in domestic ducks, showed increased virulence in ducks (Hidaka et al., 2021). The d5a20b strain had 11 amino acid substitutions in its P/V, M, F, HN, and L proteins as compared to 9a5b. In the present study, we generated a series of recombinant (r) NDVs with these amino acid substitutions to identify the molecular basis of virulence of NDV in ducks, and evaluated their influences on virulence and in vitro viral properties. Each of the single amino acid substitutions in either the F protein I142M or the M protein Q44R contributed to the enhancement of intracerebral and intranasal pathogenicity in domestic ducks. The cell-cell fusion activity of the virus with F I142M was five times higher than that of the parental r9a5b. The virus with M Q44R rapidly replicated in duck embryo fibroblasts. Additionally, the rM+F+HN strain, which has the same amino acid sequences as d5a20b in M, F, and HN proteins, showed the highest level of virulence and replication efficiency among the generated recombinant viruses, nearly comparable to rd5a20b. These results suggest that multiple factors are involved in the high growth ability of NDV in duck cells, leading to increased virulence in vivo. 相似文献
27.
Hiroshi UENO Osamu YAMATO Takeshi SUGIURA Moeko KOHYAMA Akira YABUKI Kenjiro MIYOSHI Kazuya MATSUDA Tsuyoshi UCHIDE 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2016,78(1):91-95
A male Japanese domestic cat with retarded growth in Hokkaido, Japan, showed
progressive motor dysfunction, such as ataxia starting at 3 months of age and tremors,
visual disorder and seizure after 4 months of age. Finally, the cat died of neurological
deterioration at 9 months of age. Approximately half of the peripheral blood lymphocytes
had multiple abnormal vacuoles. Magnetic resonance imaging showed bisymmetrical
hyperintensity in the white matter of the parietal and occipital lobes in the forebrain on
T2-weighted and fluid-attenuated inversion recovery images, and mild encephalatrophy of
the olfactory bulbs and temporal lobes. The activity of lysosomal acid β-galactosidase in
leukocytes was negligible, resulting in the biochemical diagnosis of GM1 gangliosidosis.
Histologically, swollen neurons characterized by accumulation of pale, slightly granular
cytoplasmic materials were observed throughout the central nervous system. Dysmyelination
or demyelination and gemistocytic astrocytosis were observed in the white matter.
Ultrastructually, membranous cytoplasmic bodies were detected in the lysosomes of neurons.
However, genetic analysis did not identify the c.1448G>C mutation, which is the single
known mutation of feline GM1 gangliosidosis, suggesting that the cat was affected with a
new variant of the feline disease. 相似文献
28.
29.
Shogo Higaki Yoshie Koyama Emi Shirai Tomoya Yokota Yasuhiro Fujioka Noriyoshi Sakai Tatsuyuki Takada 《Fish physiology and biochemistry》2013,39(3):701-711
We succeeded to establish cell lines from endemic fish species Honmoroko Gnathopogon caerulescens, which inhabits Lake Biwa, the third oldest lake in the world. Two cell lines designated as RMT1 and RMO1 were established from testis and ovary of G. caerulescens, respectively. These cell lines were initially cultured in Leibovitz’s L-15 medium supplemented with fetal bovine serum (FBS), fish embryo extract, epidermal growth factor, and basic fibroblast growth factor. Further addition of forskolin and β-mercaptoethanol was required to establish and maintain these cell lines for more than 60 passages. RMT1 and RMO1 cells showed fibroblast- and epithelial-like morphology, respectively. From immunocytochemical staining and gene expression patterns, RMT1 cells showed a characteristic of testicular Sertoli cells and RMO1 cells did that of ovarian theca cells. Both RMT1 and RMO1 cells multiplied well in the medium supplemented with 10 % FBS at 28 °C and their minimum population doubling times were 24.4 and 28.8 h, respectively. At the 45th passage, most of the RMT1 and RMO1 cells had a hyperploid set of chromosomes (67.3 and 96.1 %, respectively). Cells with normal diploid chromosome set were not observed. RMT1 cells were transfected with an enhanced green fluorescent protein (EGFP) expression vector and human elongation factor 1 α promoter worked efficiently to express EGFP. In addition, EGFP-expressing cell lines were also established, suggesting that the cell lines could be utilized as an in vitro monitor system (biosensor) for the evaluation of endocrine disruptors which might affect gonadal function. 相似文献
30.
Toshie Wakabayashi Nobuaki Suzuki Mitsuo Sakai Taro Ichii Seinen Chow 《Fisheries Science》2006,72(3):494-502
ABSTRACT: A total of 110 adult individuals from four ommastrephid (family Ommastrephidae) squid species ( Ommastrephes bartramii, Sthenoteuthis oualaniensis, Eucleoteuthis luminosa, and Hyaloteuthis pelagica ) were used to obtain diagnostic DNA markers for species identification. Restriction fragment length polymorphism (RFLP) analysis of a partial segment (855 bp) of the mitochondrial DNA cytochrome oxidase I (COI) gene amplified by polymerase chain reaction (PCR) revealed that the restriction profiles of two endonucleases ( Alu I and Tsp 509 I) were diagnostic for species identification. The restriction assay partially supplemented with nucleotide sequence analysis successfully assigned 69 damaged and morphologically equivocal ommastrephid paralarvae collected in northern Hawaiian waters, identifying 60 O. bartramii , eight S. oualaniensis , and one E. luminosa . The family Ommastrephidae appears to be monophyletic. Although the phylogenetic relationships among genera were not resolved well due to apparent homoplasy and large genetic divergence between species, COI sequence data without transitions provided support for subfamily level relationships. 相似文献