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31.
番木瓜环斑病毒株系的分子生物学方法鉴定 总被引:6,自引:1,他引:6
以PRSV株系特异性引物对PRSV的PRSV126(PRSV日本分离物)、Ys、Vb和Sm等株系进行RT-PCR方法鉴定,引物PR21/PR22能把Ys从Vb和Sm中鉴定出来,PR300/PR301则能把Vb从Ys、Sm和PRSV126中鉴定出来;用限制性内切酶Hae Ⅱ、Sau3A I和Hinf I对PRSV的PRSV126、Ys、Vb和Sm等株系进行单酶切RT-PCR-RFLP分析,Hinf I能把PRSV126与Ys、Vb和Sm鉴别开来,Sau3A I能把Ys与Vb和Sm鉴别开来,Hae Ⅱ则能把Ys与PRSV126、Vb和Sm鉴别开来;以P1/P2为引物,对Vb、Ys和Sm株系进行RT-PCR-RFLP-SSCP分析,结果能一次把三者较好地区别开来。 相似文献
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XIAO Qing-zhong LI Hao-wei WEN Guan-mei HUANG Shao-hua ZHANG Xiu-ming LI Yan LI Shu-nong 《园艺学报》2002,18(10):1179-1182
AIM: To investigate the differentiation from adult rat and human bone marrow mesenchymal stem cells (BMMSCs) into neuron with musk polypeptide (Mu-P).METHODS: Adult rat and human BMMSCs were induced with Mu-P.Neuron-specific enolase (NSE),neurofilament (NF),Nestin,glial fibrillary acidic protein (GFAP) were detected by immunohistochemistry.RESULTS: Simple methods with Mu-P induced adult rat and human BMMSCs exhibiting a neuronal phenotype,expressing Nestin at 3 hours to 5 hours,and expressing NE and NF at 5 hours to 7 days.But the neuron-like cells didn't express the glial astrocyte marker GFAP.CONCLUSION: Adult rat and human BMMSCs can be induced to differentiate into neurons with Mu-P. 相似文献
34.
玉米螟赤眼蜂适宜生境的研究与利用:Ⅲ.夏玉米间作匍匐型绿豆对赤眼蜂的增诱作用及其在穗期玉米螟防治中的利用 总被引:1,自引:0,他引:1
本文就夏玉米间作匍匐型绿豆对玉米螟赤眼蜂增诱作用及其在穗期玉米螟防治中利用的可行性进行了研究。结果表明,夏玉米间作匍匐型绿豆结合接种式放蜂区,与平作夏玉米结合接种式放蜂区和不放蜂对照区相比,明显提高了玉米螟赤眼蜂对螟卵的寄生率,1992年三者人工挂卵的卵块总寄生率分别为26.7%、3.6%和0;1993年分别为28.0%、6.6%和1.0%,自然落卵的寄生率分别为69.1%、22.4%和4.6%。1994年用心叶期抗螟品种间作绿豆结合接种式放蜂区,人工挂卵的卵块总寄生率为30.7%,而平作夏玉米不放蜂对照区仅为2.5%;自然落卵的卵块寄生率前者为56.2%,后者为12.3%。心叶期抗螟品种间作匍匐型绿豆结合接种式放蜂区、心叶期抗螟夏玉米平作不放蜂和感螟品种不放蜂对照区的平均百株蛀孔数分别为54.8、102.6和277.2个,放蜂区与两个不放蜂对照区相比,对玉米螟的防效分别为46.6%和80.2%。试验结果显示采用心叶期玉米抗螟品种间作匍匐型绿豆,并接种释放少量赤眼蜂等综合措施对穗期玉米螟害有明显的控制作用 相似文献
35.
XU Ruo-bing WEN Jian-ming ZHANG Meng LV Chang-hai XIAO Gang ZHANG Wen-min LIANG Hui-zhen 《园艺学报》2004,20(11):1982-1988
AIM: To study effects of urokinase-type plasminogen activator (uPA) signal transduction on expression of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of matrix metalloproteinase-3 (TIMP-3) in giant cell tumor of bone (GCT). METHODS: Expression of uPAR, MMP-2 and TIMP-3 in GCT tissue was detected by immunohistochemistry. Phosphorylation level of mitogen-activated protein kinase (p44) in uPA/uPAR signal pathway in cultured GCT cells was detected by immunoprecipitation. The expression of MMP-2 and TIMP-3 in cultured cells after treatment with uPA-ATF or anti-uPAR antibody was also detected by Western blotting. RESULTS: 1) Urokinase-type plasminogen activator receptor (uPAR) was positive on the cell membrane and in cytoplasm of some mononuclear stromal cells (MSCs) and multinucleated giant cells (MGCs); 2) MMP-2 was positive in the cytoplasm and on the cell membrane of almost all of MSCs and some of MGCs. The polar distribution of MMP-2 in the cytoplasm of MGCs was especially obvious; 3) The expression of TIMP-3 of some MSCs and MGCs in GCT was much lower than MMP-2. The positive signal also showed a prominent polarity; 4) After treatment with uPA-ATF, the phosphorylation level of p44 in GCT cultured cells was much higher than the control. Addition of anti-uPAR antibody in the cells remarkably down-regulated the phosphorylation level of p44 as compared with the control group, suggesting that uPA-ATF participates cell signal transduction and this reaction can be inhibited by anti-uPAR antibody; 5) uPA-ATF cell signal pathway up-regulated expression of MMP-2 and TIMP-3, while anti-uPAR antibody down-regulated the expression of MMP-2 and TIMP-3. CONCLUSION: These results demonstrate for the first time that uPA-ATF directly regulates the expression of MMP-2 and TIMP-3 by signal transduction pathway, and the over-expression of MMP-2 and TIMP-3 may play an important role in local osteolysis of GCT. 相似文献
36.
AIM: To investigate the relationship between p21WAF1gene polymorphisms and protein expression in breast carcinoma. METHODS: Polymerase chain reaction single-strand conformation polymorphisms technique (PCR-SSCP) and immunohistochemical assay of S-P immunostaining technique were used to study polymorphisms of p21WAF1 and protein expression respectively on the specimen of paraffin-embedded tissues in 100 cases of breast carcinomas and 40 benign breast diseases as control. RESULTS: Two p21WAF1 gene polymorphisms were found in 18% (18/100) of breast carcinomas and 5% (2/40) of control samples. The difference between the two groups was statistically significant (χ2=3.94, P<0.05). The positive immunohistochemical reaction of p21WAF1 protein were found in 50% (50/100) of breast carcinomas and 12.5% (5/40) of control samples. The difference between the two groups was statistically significant (χ2=16.84, P<0.01). The positive immunohistochemical reaction of p21WAF1 protein were found in 100% (18/18) of breast carcinomas with p21WAF1 gene polymorphisms and 39% (32/82) of no p21WAF1 gene polymorphisms. The difference between two groups was statistically significant (χ2=21.95, P<0.01). The p21WAF1 gene polymorphisms were correlated with the protein expression in breast carcinomas (r=0.576, P<0.01). CONCLUSION: p21WAF1 gene polymorphisms may create the different copies of mRNA and may make relevant protein molecules. 相似文献
37.
AIM: The goal of this study was to compare different methods for tumor antigen preparation, to observe the induction of tumor-specific cytotoxic T lymphocytes in rats by dendritic cells (DCs) pulsed with different tumor antigens. METHODS: The precursors of dendritic cells were isolated from bone marrow of rats, stimulated in vitro with recombinent rat granulocyte-macrophage colony-stimulating factor (rrGM-CSF) and interleukin-4 (rrIL-4). Then rat DCs were pulsed with C6 tumor cell antigens prepared with different methods: freeze-thaw, boiling or total protein extracted from ultrasonic crushed tumor cell. Subsequently primed DCs were cocultured with T lymphocytes isolated from spleen to induce CTL. Lymphocyte chemoattractant factor from DCs and cytokine IFN-γ release were determined by ELISA, the cytotoxicity of CTL was assayed by JAM test. RESULTS: DCs pulsed with boiled tumor cell in vitro induced an enhanced ability of T-cell proliferation and cytotoxic T lymphocyte activity.CONCLUSION: Our results demonstrated that DCs primed with boiled tumor cell may represent a method for inducing immune responses against the entire repertoire of tumor antigens of malignancies. 相似文献
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猪伪狂犬病基因缺失疫苗的制备、安全性、免疫原性、保存期测定及区域试验 总被引:3,自引:1,他引:3
为了提供有效的伪狂犬病疫苗,用鸡胚成纤维细胞扩大培养了PrV HB-98突变株(TK^-/gG^-/LacZ^+),研制了伪狂犬病基因缺失疫苗,并对该疫苗经肌肉接种、经口等免疫途径的最小免疫剂量进行了测定,同时也对4批疫苗的安全性、效力、免疫期和保存期进行了检测;同时将4批疫苗用于免疫23个猪场的母猪、新生仔猪和育肥猪进行区域试验。测定结果表明,疫苗经上述两种途径接种对不同阶段猪的最小免疫剂量均为10^5.0 TCID50;10倍免疫剂量的疫苗对初生仔猪、15日龄仔猪和妊娠母猪是安全的,免疫猪能抵抗强毒的攻击;疫苗在4℃和-20℃下分别可保存6个月和12个月。对伪狂犬病毒抗体阴性的70日龄商品猪和种猪的免疫期为6个月。田间试验表明,4批猪伪狂犬病基因缺失疫苗安全有效,并可用于仔猪发病时的紧急接种。为猪伪狂犬病基因工程疫苗的制备与应用提供了有力的依据。 相似文献
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