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511.
After the successful use of 3-[4,5-(dimethylthiazol-2-yl)]-2,5-diphenyltetrazolium bromide (MTT) in cell proliferation assays, its use has been established by different workers in cytotoxicity assays and research on leukaemia. In the present study, a colorimetric assay using MTT was adopted to evaluate the cytotoxic activity of chicken intestinal intraepithelial lymphocytes (iIELs), which constitute an important cellular component of the gut-associated lymphoid tissue (GALT). These iIELs are found to exhibit natural killer (NK) cell-like cytotoxic activity, which is spontaneous, non-MHC-restricted, and does not need to be primed. Hitherto, conventional chromium-release assays have been used to evaluate the cytotoxic activity of iIELs, but these assays have disadvantages such as radiation hazards and loss of the cells in washing steps. The mean percentage cytotoxic activity of chicken iIELs evaluated by the colorimetric assay was 90.37±2.53 in a group of 5-week-old chickens and 80.2±3.45 in a group of 8-week-old chickens. These findings established the successful use of a colorimetric assay using MTT for evaluating the cytotoxic activity of chickens iIELs.Abbreviations DMEM Dulbecco's modified Eagle's medium - DMSO dimethyl sulphoxide - E effector cells - GALT gut-associated lymphoid tissue - GM growth medium - iIELs intestinal intraepithelial lymphocytes - MTT 3-[4,5-(dimethylthiazol-2-yl)]-2,5-diphenyltetrazolium bromide - NK cell natural killer cell - OD optical density - RPMI Rosewell Park Memorial Institute medium - T target cells  相似文献   
512.
A mature, rumen-cannulated steer fed Coastal Bermuda grass hay for 8 weeks was adapted to a high-grain ration by stepwise increases in grain over a period of 4 weeks. The grain rations had concentrate-to-roughage ratios of 40:60, 70:30, and 85:15 and were fed for 10, 7, and 11 days, respectively. Numerical estimates of lactate-utilizing bacteria in ruminal fluid of the steer the last 3 days each ration was fed were made by colony counts. Lactate-utilizing bacteria were identified as Megasphaera elsdenii, Peptococcus asaccharolyticus, and Selenomonas ruminantium. Maximal numbers of lactate-utilizing bacteria were observed in the ruminal fluid of the steer during feeding the concentrate-to-roughage ration of 70:30.  相似文献   
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515.
Soluble nymphal antigens (HNAg) were purified by immunoaffinity chromatography using CNBr-activated Sepharose 4B coupled with immunoglobulin ligands from animals immunized with HNAg and 69–71% protected against challenge infestations, and 8% recovery of the purified protein (Aff-HNAg) was obtained. Following immunization of crossbred calves (Bos indicus×Bos taurus) with 1600 g of Aff-HNAg in three divided doses, significant rejections of larvae (p<0.001, 84.2%), nymphs (p<0.05, 61.4%) and adults (p<0.05, 58.7%) were recorded. No significant changes were recorded in the engorgement weights of the larvae and nymphs, but there was a significant (p<0.05) reduction in the weight of the engorged adults. Immunization conferred a significant decrease in the numbers of resultant nymphs (p<0.001) and adults (p<0.001) that had fed on the immunized animals. SDS-PAGE analysis identified a 39 kDa protein, previously isolated from larvae of Hyalomma anatolicum anatolicum, as the antigen responsible for the induction of resistance against all the stages of the tick.  相似文献   
516.
Breeding for resistance against Bacterial blight (BB), a destructive disease in rice, is a major challenge because of the rich diversity of the pathogen, Xanthomonas oryzae pv. oryzae. Three resistance genes i.e. xa5, xa13 and Xa21 were transferred from IRBB 60 through marker-assisted backcross breeding to supplement the Xa4 gene present in Tapaswini, an elite cultivar having a wide coverage. The precise transfer was aided through effective foreground selection using STS markers RG 556, RG 136 and pTA248 linked to the three target genes, respectively. In the screening assays, the gene pyramids having four genes exhibited higher level of resistance against the disease. Background selection based on morphological and grain quality traits and SSR markers led to the recovery of 85–96 % of the recurrent parent genome in the gene pyramids. Multi location evaluation conducted over a two year period at several hotspot locations of bacterial blight in India has resulted in identification and release of CRMAS 2622-7-6 (IT-6), a promising gene pyramid as improved Tapaswini. The high levels of resistance exhibited in the improved genotype without any yield penalty and its similarity to the recurrent parent provides a successful demonstration of the potential of MAS in pyramiding genes as well as the recovery of the parental genome with high accuracy.  相似文献   
517.
Antimicrobial activity of crude rhizome oils from some important members of Zingiberaceae from Northeast India was carried out in the present study. The plants used were Curcuma amada, C. longa, Zingiber moran, and Z. zerumbet. Rhizomes of the four species were collected and rhizome oil was extracted using six different polar and non-polar solvents. The antimicrobial property of the crude oil was studied against human pathogens using disc-diffusion and viability assay. The rhizome oil of all the 24 extracts exhibited potent antimicrobial activity against all pathogenic bacterial and fungal strains tested. Hydrodistilled fractions of the four species studied showed better effect among all solvents. Water extract of Z. moran was found to be the highest effective antimicrobial agent of all. Antibacterial effect of the water fractions was also characterized against Gram-positive and Gram-negative bacteria using micro Raman spectroscopy. This investigation shows that the essential oils from the wild and domestic species of the family Zingiberaceae from Northeast India are a potential source of bioactive compounds with tremendous medicinal, economic, and therapeutic value. The wild and endemic species Z. moran represents an interesting and promising source of plant-derived antimicrobial agent in the development of new drugs against pathogens.  相似文献   
518.
Pharmacologic inhibition of autophagy can be achieved using lysosomotropic agents such as hydroxychloroquine (HCQ) that interfere with fusion of the autophagosome to the lysosome thus preventing completion of the recycling process. The goal of the present study is to determine the sensitivity of eight canine (cOSA) and four human (hOSA) osteosarcoma tumour cell lines to antiproliferative and cytotoxic effects of lysosomal autophagy inhibitors, and to compare these results to the autophagy-dependence measured using a CRISPR/Cas9 live-cell imaging assay in OSA and other tumour cell lines. Antiproliferative and cytotoxic response to HCQ and Lys05 was determined using live cell imaging and YOYO-1 staining. CRISPR/Cas9 live cell imaging screen was done using species specific guide RNA's and transfection of reagents into cells. Response to autophagy core genes was compared to response to an essential (PCNA) and non-essential (FOXO3A) gene. cOSA and hOSA cell lines showed similar antiproliferative and cytotoxic responses to HCQ and Lys05 with median lethal dose (Dm) values ranging from 4.6–15.8 μM and 2.1–5.1 μM for measures of anti-proliferative response, respectively. A relationship was observed between antiproliferative responses to HCQ and Lys05 and VPS34 CRISPR score with Dm values correlating with VPS34 response (r = 0.968 and 0.887) in a species independent manner. The results show that a subset of cOSA and hOSA cell lines are autophagy-dependent and sensitive to HCQ at pharmacologically-relevant exposures.  相似文献   
519.
Recent studies in the United States reported that approximately 85% of pre-weaned dairy calves were infected with zoonotic Cryptosporidium parvum, whereas only 1-2% of post-weaned calves and 1-2-year-old heifers were infected with this species. Cryptosporidium bovis and Cryptosporidium deer-like genotype were much more prevalent in the post-weaned animals. It is not clear whether the same infection pattern also occurs in other geographic areas. In this study, to determine whether the same Cryptosporidium infection pattern was present in other geographic areas, we genotyped Cryptosporidium specimens collected from two farms in China and India, using specimens from farms in Georgia, USA for comparison. C. bovis was the most common species found in pre- and post-weaned calves in all three areas. In Georgia, the deer-like genotype was found frequently in pre- and post-weaned calves and Cryptosporidium andersoni was found in one post-weaned calf. Both C. bovis and the deer-like genotype were found in the few milking cows examined in Georgia. There were no differences in the small subunit rRNA gene sequences obtained from C. bovis or deer-like genotype among the three areas. One adult yak in China, however, was infected with a species similar to C. bovis, with only three nucleotide mutations in the target gene. All four common bovine Cryptosporidium spp. were differentiated from each other by restriction fragment length polymorphism analysis of PCR products with enzymes SspI and MboII. Thus, both C. bovis and the deer-like genotype are found in all age groups of cattle in diverse geographic areas and host adaptation of C. bovis might have occurred in yaks.  相似文献   
520.
Journal of Pest Science - The fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith), is a highly destructive insect pest of several crop plants and threatening global food security. The current...  相似文献   
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