首页 | 本学科首页   官方微博 | 高级检索  
文章检索
  按 检索   检索词:      
出版年份:   被引次数:   他引次数: 提示:输入*表示无穷大
  收费全文   6994篇
  免费   399篇
  国内免费   754篇
林业   161篇
农学   430篇
基础科学   19篇
  260篇
综合类   2058篇
农作物   299篇
水产渔业   463篇
畜牧兽医   2939篇
园艺   1446篇
植物保护   72篇
  2024年   17篇
  2023年   75篇
  2022年   209篇
  2021年   272篇
  2020年   265篇
  2019年   361篇
  2018年   204篇
  2017年   308篇
  2016年   341篇
  2015年   325篇
  2014年   375篇
  2013年   419篇
  2012年   517篇
  2011年   530篇
  2010年   323篇
  2009年   322篇
  2008年   347篇
  2007年   500篇
  2006年   334篇
  2005年   273篇
  2004年   270篇
  2003年   201篇
  2002年   168篇
  2001年   140篇
  2000年   118篇
  1999年   90篇
  1998年   79篇
  1997年   82篇
  1996年   81篇
  1995年   70篇
  1994年   96篇
  1993年   86篇
  1992年   64篇
  1991年   54篇
  1990年   48篇
  1989年   42篇
  1988年   38篇
  1987年   23篇
  1986年   18篇
  1985年   8篇
  1984年   7篇
  1983年   3篇
  1982年   6篇
  1981年   7篇
  1980年   9篇
  1979年   5篇
  1978年   2篇
  1975年   3篇
  1973年   2篇
  1956年   8篇
排序方式: 共有8147条查询结果,搜索用时 15 毫秒
1.
In this study, primary and immortalized bovine intestinal epithelial cells (BIECs) were characterized for the expression of surface carbohydrate moieties. Primary BIEC-c4 cells showed staining greater than 90 % for 16 lectins but less than 50 % staining for four lectins. Immortalized BIECs showed significantly different lectin binding profile for few lectins compared to BIEC-c4 cells. BIEC-c4 cells were studied for infectivity to E. coli, Salmonella enterica, bovine rotavirus, bovine coronavirus, and bovine viral diarrhea virus. Bovine strain E. coli B41 adhered to BIEC-c4 cells and Salmonella strains S. Dublin and S. Mbandaka showed strong cell invasion. BIEC-c4 cells were susceptible to bovine rotavirus. LPS stimulation upregulated IL-10, IL-8, and IL-6 expression and Poly I:C upregulated TLR 8 and TLR 9 expression. This study provides important knowledge on the glycoconjugate expression profile of primary and immortalized BIECs and infectivity and immune responses of primary BIECs to bacterial and viral pathogens or ligands.  相似文献   
2.
Equine herpesvirus type 1 (EHV-1) is responsible for respiratory disorders, abortion and myeloencephalopathy (EHM) in horses. Two pathotypes of EHV-1 strains are circulating in the field: neurovirulent (N) and non-neurovirulent (NN). For both strains, CD172a+ monocytic cells are one of the main carrier cells of EHV-1 during primary infection, allowing the virus to invade the horse’s body. Recently, we showed that EHV-1 NN strains showed a restricted and delayed replication in CD172a+ cells. Here we characterize the in vitro replication kinetics of two EHV-1 N strains in CD172a+ cells and investigate if the replication of these strains is similarly silenced as shown for EHV-1 NN strains. We found that EHV-1 N replication was restricted to 7–8% in CD172a+ cells compared to 100% in control RK-13 cells. EHV-1 N replication was not delayed in CD172a+ cells but virus production was significant lower (103.0 TCID50/105 inoculated cells) than in RK-13 cells (108.5 TCID50/105 inoculated cells). Approximately 0.04% of CD172a+ cells produced and transmitted infectious EHV-1 to neighbour cells compared to 65% of RK-13 cells. Unlike what we observed for the NN strain, pretreatment of CD172a+ cells with histone deacetylases inhibitors (HDACi) did not influence the replication of EHV-1 N strains in these cells. Overall, these results show that the EHV-1 replication of N strains in CD172a+ cells differs from that observed for NN strains, which may contribute to their different pathogeneses in vivo.  相似文献   
3.
4.
Two-cell stage and blastocyst stage mouse embryos were equilibrated in a medium containing 7.5% ethylene glycol (EG) and 7.5% dimethyl sulfoxide (DMSO) for 8–15 min. Vitrification was performed in a medium containing 0.5 M sucrose and either 15% EG + 15% DMSO, 17.5% EG + 17.5% DMSO, or 20% EG + 20% DMSO for 30 s. They were then placed either on a hemi-straw (HS) or a hollow fiber vitrification (HFV) device and vitrified by cooled air inside a 0.5-ml straw. In two-cell embryos, a 100% survival rate was obtained from all groups except the 20% HS group (P > .05). All vitrified two-cell groups showed similar rates of blastocyst development to that of fresh control group (P > .05), except 17.5% and 20% HFV groups, which were significantly lower than the other groups (P < .05). In the blastocyst embryos, the HFV groups were divided into two subgroups (non-collapsed; HFV-NC and collapsed; HFV-C blastocyst). Re-expansion rate in 15% HFV-NC, 17.5% HFV-NC, and 15% HFV-C groups was reduced (P < .05), whereas the rest were similar to control. In conclusion, we established a simplified, reliable, and closed system for HFV vitrification applying hemi-straw, which does not require skilled practitioners.  相似文献   
5.
对5个无核葡萄品种的试验表明,不同培养方式对离体胚发育的影响是很明显的,其中4个品种胚发育率液体培养高于固体培养,‘京早晶’在液体培养中胚发育率高达73.7%,是目前国内外胚发育率最高的报道;液体培养比固体培养胚的发育程度高,发育整齐,无畸形胚,易成苗.  相似文献   
6.
波尔山羊胚胎数量、质量与受体移植妊娠率的关系   总被引:4,自引:0,他引:4  
本文就波尔山羊胚胎数量,质量与受体移植妊娠率的关系进行了探讨。结果表明:胚胎质量优劣是直接影响胚胎移植成功率的关键因素。移植胚胎数量因品种不同有差异。移植双胚,奶山羊受体胚胎成羔率53.4%,明显高于成都麻羊胚胎成羔率36.3%,成都麻羊单胚成羔率为66.7%。  相似文献   
7.
牛皮肤成纤维细胞的体外培养与冻存   总被引:10,自引:0,他引:10  
利用牛皮肤组织块直接培养法,得到牛皮肤细胞的原代培养物,再用酶消化法和反复贴壁法处理,能够纯化成纤维细胞。成纤维细胞的冻存是通过选用6种分别含有二甲基亚砜(DMSO)、甘油(GL)及乙二醇(EG)的保护液,以相同的冻前处理方法,对牛皮肤成纤维细胞进行缓慢冷冻,冰箱预冷平衡1-2h,逐步投入液氮(-196℃)中保存,再经37℃水浴解冻,Hanks液脱保护剂,以贴壁率评价冻存效果。结果表明,20%DMSO保护液对牛皮肤成纤维细胞表现出较好且稳定的冷冻保护效果,其平均贴壁率达87.9%。  相似文献   
8.
Kano  R.  Kubota  A.  Nakamura  Y.  Watanabe  S.  Hasegawa  A. 《Veterinary research communications》2001,25(8):615-622
Using cDNA from a CRFK cell line as a template, PCR amplification was performed with the Ub1S and poly(dT) primers to isolate feline ubiquitin genes. Sequencing of the 495 bp PCR fragment revealed that the putative amino acids induced by this fragment gave a fusion protein consisting of a ubiquitin polypeptide (76 amino acids) and an extension protein of ribosomal proteins L40 (52 amino acids). The putative amino acid sequence of ubiquitin was identical to those of humans, rats and pigs.The recombinant glutathione S-transferase (GST)–feline ubiquitin fusion proteins were produced in Escherichia coli and purified. The fusion proteins had a molecular weight of about 42 kDa and were detected by immunoblot assay with rabbit anti-ubiquitin antiserum.The mRNAs from heat-shocked and non-heat-shocked cells were subjected to RT-PCR (Ub1S and poly(dT) primers) analysis. The molecular weights of the ubiquitinated proteins in heat-shocked CFRK cells were between 18 kDa and 24 kDa by immunoblot assay.These results suggested that there were more ubiquinated proteins in the heat-shocked CRFK cells than in the pre-heat-shocked cells.  相似文献   
9.
桑悬浮细胞原生质体培养的研究   总被引:2,自引:1,他引:1  
陈爱玉  王勇 《蚕业科学》1993,19(3):135-138
采用继代培养三个月后的桑子叶悬浮细胞为材料,进行了原生质体的分离和培养。桑悬浮细胞在纤维素酶、果胶酶、半纤维素酶的混合溶液中,酶解获得产量高、活力强的原生质体。原生质体在K8p(附加6—BA、NAA、2,4—D、LH)液体培养基中,再生细胞经多次分裂,得到肉眼可见的小愈伤组织。再通过增殖继代培养,获得浅黄色、具有明显颗粒结构的愈伤组织,转至各种激素含量的MSB固体培养基中,尚未获得绿苗分化。  相似文献   
10.
鸭病毒性肝炎鸭胚灭活疫苗的研究   总被引:6,自引:0,他引:6  
以鸭病毒性肝炎病毒Ⅰ型ATCC毒株接种健康鸭胚,收集致死的全胚组织作制苗材料,用福尔马林灭活,以麸氨酸钠终止其作用。先后试制疫苗11批,在试验室免疫雏鸭71只,经强毒攻击后,总的保护率为90.4%。本疫苗免疫雏鸭3天后可产生较坚强的免疫力,在室温至少可保存11天,在4℃可保存254天。经在江苏、安徽、广东等省推广应用200000头剂,均安全有效,颇受用户欢迎,对控制鸭病毒性肝炎的流行起了重要作用。  相似文献   
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号