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在制备了两个Cell Ⅰ-Hep Ⅱ 双结构域重组FN多肽(CH50和CH56)的基础上,研究其抑制肿瘤细胞浸润能力的作用。两个多肽的结构差异是CH50中删除了Cell I和HepⅡ之间的Ⅲ-11和ED-A结构顺序。CH50(ED_(50)为30.2 nmol/L)结合细胞的能力略高于CH56(ED_(50)为45.4 nmol/L)。两种多肽均可显著抑制黑色素瘤B16/F1细胞结合层粘素的能力,抑制作用相同。在体内肿瘤浸润抑制试验中,两种多肽均可显著抑制癌细胞浸润能力,使肺转移结节数降低80%左右。结果提示:Ⅲ-11和ED-A结构顺序对Cell Ⅰ-Hep Ⅱ 双结构域多肽结合细胞的能力有一定的影响,但删除Ⅲ-11和ED-A不是重组多肽抑制肿瘤转移的决定因素,Cell I和Hep Ⅱ 这两个结构域单独连接在一起是其抑制肿瘤细胞转移的结构基础。 相似文献
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Katsuo Kamata Akemi Sakamoto Yutaka Kasuya 《Naunyn-Schmiedeberg's archives of pharmacology》1988,338(4):401-406
Summary The characteristics of the non-adrenergic, noncholinergic inhibitory response of the rat stomach fundus to transmural nerve stimulation were compared with the relaxation induced by vasoactive intestinal polypeptide (VIP). Treatment with -chymotrypsin (5 U/ml) or VIP antiserum (1:200) significantly reduced the relaxation induced by transmural nerve stimulation at 30 Hz, indicating that the possible transmitter in the non-adrenergic, non-cholinergic nerves is a peptide and may be VIP or a closely related peptide. VIP was able to relax, fully and dose-dependently, the stomach fundus that had previously been constricted by treatment with 10–6 M serotonin, and the IC50 value for VIP was 2.4 × 10–9 M. VIP elevated levels of cyclic AMP in a dose-dependent manner and the EC50 value was 2.8 × 10–9 M in the presence of 10–6 M atropine and 10–6 M guanethidine. The stomach fundus was relaxed by transmural nerve stimulation (30 Hz, 50 mA) and transmural nerve stimulation also caused production of cyclic AMP in the rat stomach in the presence of atropine and guanethidine. The basal level of cyclic AMP in the stomach was 8.7 ± 0.26 pmole/mg protein. When transmural nerve stimulation was applied for 5 min, the contraction of the stomach, induced by 10–6 M serotonin, was inhibited by 54% in the presence of atropine and guanethidine and the level of cyclic AMP was increased to 13.0 ± 0.73 pmol/mg protein. Apamin inhibited the transmural nerve stimulation-induced relaxation and shifted the dose-response curve for VIP to the right. These results suggest that one of the putative neurotransmitter from non-adrenergic, non-cholinergic nerves in the rat stomach is VIP and that VIP-induced relaxation may be mediated by the production of cyclic AMP and by the opening of apamin-sensitive K+-channels.Send offprint requests to K. Kamata at the above address 相似文献
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目的 观察血管活性肠多肽(VIP)对老龄勃起功能障碍(ED)大鼠的作用及其机制。方法40只SPF级雄性SD大鼠饲养至18月龄构建老龄模型大鼠,采用阿扑吗啡实验筛选老龄ED大鼠,阿扑吗啡实验结果阳性的老龄勃起功能正常大鼠为正常组,实验结果阴性的老龄ED大鼠为ED组和干预组,每组7只。干预组大鼠使用VIP 25 ng/kg隔日腹腔注射,治疗28 d。检测阴茎海绵体内压(ICP)及平均动脉压(MAP)评估勃起功能;酶联免疫吸附法检测大鼠阴茎海绵体组织环磷酸腺苷(cAMP)、一氧化氮(NO)含量;组织免疫荧光技术(IF)测大鼠阴茎海绵体组织血管性血友病因子(vWF)表达水平;蛋白质印迹法检测海绵体蛋白激酶A (PKA)、内皮源性一氧化氮合酶(eNOS)、血管性血友病因子(vWF)及血管内皮生长因子(VEGF)蛋白表达水平。结果 正常组、ED组和干预组基础ICP分别为(20.41±5.92)mmHg、(21.76±5.37)mmHg和(18.54±3.97)mmHg(1mmHg=0.133 kPa),MAP分别为(123.52±14.74)mmHg,(118.83±10.97)mmHg和(114... 相似文献
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Carlo Alberto Maggi Riccardo Patacchini Paolo Santicioli Sandro Giuliani Damiano Turini Gabriele Barbanti Patrizia Beneforti Daniele Misuri Alberto Meli 《Naunyn-Schmiedeberg's archives of pharmacology》1989,339(4):415-423
Summary (1) Longitudinal muscle strips from the human small intestine (jejunum/ileum) responded to electrical field stimulation (1–50 Hz) with frequency-related primary contractions which were largely atropine- (3 M) sensitive. When the tone was raised by addition of galanin (0.3 – 1 M), prostaglandin (PG) E2 (1–10 M) or neurokinin A (NKA, 0.1 M), a frequency-related relaxation was evident which was potentiated by atropine. All the responses to field stimulation were abolished by tetrodotoxin (1 M), thus indicating their neural origin. (2) The atropine-sensitive primary contraction to field stimulation was virtually abolished by omega conotoxin fraction GVIA (CTX, 0.1–0.3 M) while the relaxations were CTX-resistant. The field stimulation-induced relaxations, which were observed in the presence of atropine and guanethidine (3 M), were also unaffected by apamin (0.1 M). (3) NKA and substance P (SP) produced a concentration- (1 nM–1 M for both peptides) related contraction, NKA being about 53 times more potent than SP. [Pro9]SP sulphone and [MePhe7]-NKB, selective agonists of the NK-1 and NK-3 receptor, respectively, were barely effective. On the other hand, [\Ala8]NKA(4–10), a selective NK-2 receptor agonist, had a potent contractile activity, similar to that of NKA. (4) Galanin (1 nM–1M) produced an atropine- and tetrodotoxin-resistant concentration-related contraction of longitudinal muscle of human isolated small intestine. The response to galanin did not show any sign of fading and was particularly suitable to study the evoked relaxations. (5) Calcitonin gene-related peptide (CGRP) (10–100 nM) consistently inhibited the nerve-mediated contractions of strips from the ileum while the effect on the jejunum was less pronounced. Vasoactive intestinal polypeptide (VIP, 0.1–1 M) inhibited nerve-mediated contractions both in the ileum and the jejunum. (6) These experiments indicate that both cholinergic excitatory and non-adrenergic non-cholinergic nerves affect motility of the longitudinal muscle of the human small intestine. Furthermore, several neuropeptides produce potent motor effects, the contractile response to tachykinins being apparently mediated by activation of NK-2 receptors. 相似文献
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H. Schwörer K. Racké H. Kilbinger 《Naunyn-Schmiedeberg's archives of pharmacology》1989,339(5):540-545
Summary Isolated segments of the guinea pig small intestine were vascularly perfused and the release of endogenous serotonin (5-HT) and its metabolite 5-hydroxyindoleacetic acid (5-HIAA) into the portal vein was measured. All test substances were intraarterially perfused. Vasoactive intestinal polypeptide (VIP, 1 pmol/l — 100 nmol/1) inhibited the spontaneous release of 5-HT and 5-HIAA. The maximal inhibitory effect (about 60%) was seen at 100 pmol/1. The effect of VIP on the spontaneous release of 5-HT and 5-HIAA was not changed in the presence of 1 ol/l tetrodotoxin (TTX).Raising intraluminal pressure by 500 Pa for 5 min increased the release of 5-HT and 5-HIAA by about 25%. Raising the intraluminal pressure in the presence of VIP reduced the release of 5-HT and 5-HIAA by about 75%. In the presence of TTX (1 gmol/l), raising intraluminal pressure also caused a decrease of the release of 5-HT and 5-HIAA which was unaffected by the additional presence of VIP. The fluid volume expelled during peristaltic activity was not affected by VIP, but reduced by about 90% in the presence of TTX.In conclusion the results demonstrate a direct inhibitory effect of VIP on the release of 5-HT from the enterochromaffin cells. In addition, VIP appears to interfere with the neuronally mediated stimulation of 5-HT release during peristaltic activity.
Send offprint request to H. Schwörer at the above address 相似文献
19.
Differential increases in brain levels of neuropeptide Y and vasoactive intestinal polypeptide after kainic acid-induced seizures in the rat 总被引:7,自引:0,他引:7
Josef Marksteiner Günther Sperk Dagmar Maas 《Naunyn-Schmiedeberg's archives of pharmacology》1989,339(1-2):173-177
Summary Changes in immunoreactivities of neuropeptide Y (NPY) and vasoactive intestinal polypeptide (VIP) were investigated in the brain of rats after severe kainic acid (KA, 10 mg/kg, i.p.) induced limbic seizures. Decreased levels of both neuropeptides were observed in the frontal cortex, striatum, dorsal hippocampus and amygdala/pyriform cortex subsequently to the period of acute seizures (3 h after injection of the toxin). Then NPY increased consistently in the frontal cortex, hippocampus and amygdala/pyriform cortex. Highest levels (290% of controls) were found in the frontal cortex after two months. Anticonvulsant therapy with phenobarbital (20 mg/kg, i.p., twice daily for three weeks) partially suppressed the rise in NPY levels. Immunoreactivity of VIP increased (to 150%) in the frontal cortex only transiently 3 days after injection of kainic acid. At the subsequently examined time intervals (10–60 days after kainic acid) it declined to control values. Levels decreasing subsequently to acute seizures reflect increased release and degradation of the respective peptide. Increased NPY levels suggest upregulation of NPY/ somatostatin/GABA neurons due to the decreased seizure threshold of the animals. The early, reversible rise of VIP in the cortex points to a short-lasting activation of this peptide system contained in local cholinergic neurons. This may be a consequence either of the acute seizures or subsequent neuropathological changes.
Send offprint requests to G. Sperk at the above address 相似文献
20.
本实验系应用荧光免疫组织化学的方法观察猴下位腰段椎旁交感神经节(L_(6-7))中神经肽Y,血管活性肠肽,降钙素基因相关肽,和P物质的存在、分布情况以及它们与酪氨酸羟化酶的共存关系。结果表明,大量细胞呈神经肽Y免疫反应阳性,它们在神经节周边分布更为密集。中等数量的血管活性肠肽阳性细胞和小量降钙素基因相关肽细胞散在于神经节内。在经含有Colchiciue的培养液离体孵育12h的标本上,可见中等数量的P物质免疫反应阳性细胞。根据抗酪氨酸羟化酶(TH)抗体的免疫染色结果,神经节内的神经元可分为TH~+和TH~-两群,前者占大多数。相邻切片免疫染色结果表明,几乎所有神经肽Y免疫阳性细胞同时含有TH,而所有血管活性肠肽免疫反应阳性细胞均呈酪氨酸羟化酶免疫反应阴性。神经肽Y与血管活性肠肽无共存关系。降钙素基因相关肽存在于部分血管活性肠肽免疫反应阳性细胞中,即属于VIP~+/TH~-组。从以上结果得出结论,在猴下位腰段椎旁交感神经节中,神经肽Y与血管活性肠肽分别存在于TH~+和TH~-两个细胞群。即神经肽Y存在于TH阳性神经元中,血管活性肠肽和降钙素基因相关肽则存在于TH阴性神经元中。 相似文献