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Quantal evoked depolarizations underlying the excitatory junction potential of the guinea-pig isolated vas deferens

JOURNAL OF PHYSIOLOGY-LONDON 1999年第2期520卷 527-537页

作者： Manchanda, R Venkateswarlu, K Indian Inst Technol Sch Biomed Engn Bombay 400076 Maharashtra India

1. The effects of a putative gap junction uncoupling agent, heptanol, on the intracellularly recorded junction potentials of the guinea-pig isolated ras deferens have been investigated. 2. After the stimulation-evoked excitatory junction potentials (EJPs) had been suppressed by heptanol (2.0 mM) to undetectable levels, a different pattern of evoked activity ensued. This consisted of transient depolarizations: that were similar to EJPs in being stimulus locked and in occurring at a fixed latency, but differed from EJPs in that they occurred intermittently and had considerably briefer time courses. 3. Analysis of the amplitudes and temporal parameters of the rapid residual depolarizations revealed a close similarity with spontaneous EJPs (SEJPs). There was no statistically significant difference between the rise times, time constants of decay and durations of the rapid residual depolarizations and of SEJPs. 4. Selected evoked depolarizations were virtually identical to SEJPs occurring in the same cell. Evoked depolarizations of closely similar amplitude and time course also occurred, usually within a few stimuli of each other. 5. These depolarizations appear to represent the individual quantal depolarizations that normally underlie the EJP and are therefore termed 'quantal excitatory junction potentials' (QEJPs) to distinguish them from both EJPs and SEJPs. 6. We examined the possibility that heptanol revealed QEJPs by disrupting electrical coupling between cells in the smooth muscle syncytium. Heptanol (2.0 mM) had no effect on the amplitude distribution, time courses, or the frequency of occurrence of SEJPs. Intracellular input impedance (R-in) of smooth muscle cells was left unaltered by heptanol. 7. 'Cable' potentials of the vas deferens, recorded using the partition stimulation method, also remained unchanged in the presence of heptanol. Thus, heptanol appeared not to modify syncytial electrical properties of the smooth muscle in any significant way. 8. Our obser

关键词：电生理学缝隙接合部/药物作用庚醇/药理学肌平滑血管/药物作用神经肌肉去极化药/药理学神经肌肉接头/药物作用突触传递/药物作用输精管/药物作用动物豚鼠男(雄)性

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Cadmium inhibits vacuolar H⁺ATPase-mediated acidification in the rat epididymis

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BIOLOGY OF REPRODUCTION 2000年第2期63卷 599-606页

作者： Herak-Kramberger, CM Sabolic, I Blanusa, M Smith, PJS Brown, D Breton, S Inst Med Res & Occupat Hlth Unit Mol Toxicol Zagreb 10001 Croatia Inst Med Res & Occupat Hlth Mineral Metab Unit Zagreb 10001 Croatia Marine Biol Lab BioCurrents Res Ctr Woods Hole MA 02543 USA Massachusetts Gen Hosp Renal Unit Boston MA 02129 USA Massachusetts Gen Hosp Program Membrane Biol Boston MA 02129 USA Harvard Univ Sch Med Dept Pathol Boston MA 02215 USA Harvard Univ Sch Med Dept Pathol Boston MA 02215 USA

In rats, an acidic luminal pH maintains sperm quiescence during storage in the epididymis. We recently showed that vacuolar H(+)ATPase-rich cells in the epididymis and vas deferens are involved in the acidification of these segments. Treatment of rats with cadmium (Cd) leads to alkalinization of this fluid by an unknown mechanism. Because Cd may affect H(+)ATPase function, we examined 1) the in vivo effect of Cd poisoning on H(+)ATPase-rich cell morphology and on the abundance and distribution of the 31-kDa H(+)ATPase subunit in cells along the rat epididymis, and 2) the in vitro effect of Cd on H(+)ATPase activity and function in the isolated vas deferens. Immunofluorescencce and immunoblotting data from rats treated with Cd for 14-15 days (2 mg Cd/kg body mass/day) showed that 1) H(+)ATPase-positive cells regressed to a prepubertal phenotype, and 2) H(+)ATPase was lost from the apical pole of the cell and was redistributed into an intracellular compartment. In experiments in vitro, Cd inhibited bafilomycin-sensitive ATPase activity in isolated total cell membranes and, as measured using a proton-selective extracellular microelectrode, inhibited proton secretion in isolated vas deferens. We conclude that alkalinization of the tubule fluid in the epididymis and vas deferens of Cd-treated rats may result from the loss of functional H(+)ATPase enzyme in the cell apical domain as well as from a direct inhibition of H(+)ATPase function by Cd.

关键词：印迹法蛋白质镉/药理学酶抑制剂/药理学附睾/酶学荧光抗体技术氢离子浓度免疫组织化学质子转运ATP酶类/分析质子转运ATP酶类/拮抗剂和抑制剂质子转运ATP酶类/代谢质子大鼠 Wistar 性成熟空泡/酶学输精管/药物作用输精管/代谢动物男(雄)性大鼠

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Multiple cellular mechanisms mediate the effect of lobeline on the release of norepinephrine

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JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS 2000年第1期294卷 302-307页

作者： Sántha, E Sperlágh, B Zelles, T Zsilla, G Tóth, PT Lendvai, B Baranyi, M Vizi, ES Hungarian Acad Sci Inst Expt Med Dept Pharmacol H-1450 Budapest Hungary

The complex effect of lobeline on [H-3] norepinephrine ([H-3]NE) release was investigated in this study. Lobeline-induced release of [H-3]NE from the vas deferens was strictly concentration-dependent. In contrast, electrical stimulation-evoked release was characterized by diverse effects of lobeline depending on the concentration used: at lower concentration (10 mu M), it increased the release and at high concentration (100 and 300 mu M), the evoked release of [H-3]NE was abolished. The effect of lobeline on the basal release was [Ca2+]-independent, insensitive to mecamylamine, a nicotinic acetylcholine receptor antagonist, and to desipramine, a noradrenaline uptake inhibitor. However, lobeline-induced release was temperature-dependent: at low temperature (12 degrees C), at which the membrane carrier proteins are inhibited, lobeline failed to increase the basal release. Lobeline dose dependently inhibited the uptake of [H-3]NE into rat hippocampal synaptic vesicles and purified synaptosomes with IC50 values of 1.19 +/- 0.11 and 6.53 +/- 1.37 mu M, respectively. Lobeline also inhibited Ca2+ influx induced by KCl depolarization in sympathetic neurons measured with the Fura-2 technique. In addition, phenylephrine, an alpha(1)-adrenoceptor agonist, contracted the smooth muscle of the vas deferens and enhanced stimulation-evoked contraction. Both effects were inhibited by lobeline. Our results can be best explained as a reversal of the monoamine uptake by lobeline that is facilitated by the increased intracellular NE level after lobeline blocks vesicular uptake. At high concentrations, lobeline acts as a nonselective Ca2+ channel antagonist blocking pre- and postjunctional Ca2+ channels serving as a counterbalance for the multiple transmitter releasing actions.

关键词：钙/代谢钙通道/药物作用地昔帕明/药理学洛贝林/药理学肌收缩/药物作用烟碱激动剂/药理学去甲肾上腺素/分泌输精管/药物作用输精管/生理学输精管/分泌动物豚鼠男(雄)性大鼠

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Subsensitivity to opioids is receptor-specific in isolated guinea pig ileum and mouse vas deferens after obstructive cholestasis

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JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS 2000年第3期293卷 946-951页

作者： Dehpour, AR Rastegar, H Jorjani, M Roushanzamir, F Joharchi, K Ahmadiani, A Univ Tehran Med Sci Sch Med Dept Pharmacol Tehran Iran Shaheed Beheshti Univ Med Sci Sch Med Dept Pharmacol Tehran Iran

The rate and degree of subsensitivity development to morphine (mu-opioid receptor, preferred, but not selective agonist) and U50488H (highly selective kappa-opioid receptor agonist) were assessed in vitro on guinea pig ileum (GPI) of cholestatic animals 2, 5, and 7 days after bile duct ligation. In addition to this phenomenon of morphine, the effects of U50488H and SNC 80 (highly selective delta-opioid receptor agonist) were studied in vitro on mice vas deferens (MVD) of cholestatic animals 2, 5, 7, 10, and 15 days after bile duct ligation. The IC50 for each compound was determined in these preparations. The ratio of the IC50 in bile duct-ligated animals to sham and control animals provides a quantitative index for the degree of subsensitivity development to each agonist. For any given time, the highest degree of subsensitivity to morphine was observed in GPI of cholestatic animals, whereas in MVD obtained from the cholestatic animals, the highest degree of subsensitivity developed to inhibitory effect of SNC 80. The subsensitivity development in cholestatic animals was time dependent;in GPI the maximum subsensitivity developed after 7 days of the operation, whereas the maximum subsensitivity in MVD developed 15 days after bile duct ligation. Moreover, subsensitivity to exogenous acetylcholine and norepinephrine in GPI and MVD, respectively, did not develop in the presence of subsensitivity to opioids in cholestatic animals. Significant accumulation of endogenous opioids in plasma of cholestatic animals has been shown in several studies and this may account for a significant development of subsensitivity to inhibitory effects of opioid agonists.

关键词：苯甲酰胺类/药理学胆汁淤积/病理生理学剂量效应关系药物回肠/药物作用回肠/生理学吗啡/药理学肌收缩/药物作用麻醉品/药理学哌嗪类/药理学受体阿片样/分类受体阿片样/生理学时间因素输精管/药物作用输精管/生理学动物豚鼠男(雄)性小鼠

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Differential blockade of α,β-methylene-ATP, noradrenaline and electrically evoked contractions of the rat vas deferens

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JOURNAL OF AUTONOMIC PHARMACOLOGY 1999年第4期19卷 249-256页

作者： Boselli, C Grana, E Univ Pavia Fac Farm Ist Farmacol Pavia Italy

1 The present study was carried out compare the effects of nifedipine, verapamil, papaverine and chloroethylclonidine (CEC) on the electrically-induced contractions and on the contractions produced by exogenous noradrenaline and alpha,beta methylene-ATP in the epidydimal portion of rat vas deferens, 2 Nifedipine inhibited in a concentration-dependent fashion the purinergic component (phase I) of the electrically evoked response. Verapamil (10(-7) M-10(-5) M) did not inhibit significantly or even potentiated phase I of the contractile response to single-pulse field stimulation but inhibited the response to alpha,beta methylene-ATP. Papaverine left unaffected phase I but depressed the response to alpha,beta methylene-ATP. CEC significantly potentiated,the purinergic component of the electrically-evoked response and the response induced by alpha,beta methylene-ATP. 3 Nifedipine was devoid of any inhibitory action on the noradrenergic component (phase II) of the response to single shock. Verapamil at the highest concentration used (10(-5) M) was able to partially inhibit phase II. Papaverine abolished in a concentration-dependent manner the noradrenergic component of the response to single shock. CEC abolished the phase II of single shock while was devoid of any inhibitory action on exogenous noradrenaline. 4 The implications of these differences among the compounds studied in the present work are discussed in relation to roles of calcium channels.

关键词：肾上腺素能α拮抗剂/药理学可乐定/类似物和衍生物可乐定/药理学色满卡林/药理学药物相互作用电刺激硝苯地平/药理学去甲肾上腺素/拮抗剂和抑制剂去甲肾上腺素/药理学罂粟碱/药理学大鼠 Wistar 输精管/药物作用血管收缩药/拮抗剂和抑制剂血管收缩药/药理学血管舒张药/药理学维拉帕米/药理学动物男(雄)性大鼠

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SYNTHESIS AND OPIOID ACTIVITIES OF STEREOISOMERS AND OTHER D-AMINO-ACID ANALOGS OF METHIONINE-ENKEPHALIN

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BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 1976年第3期73卷 632-638页

作者： COY, DH KASTIN, AJ SCHALLY, AV MORIN, O CARON, NG LABRIE, F WALKER, JM FERTEL, R BERNTSON, GG SANDMAN, CA UNIV LAVAL CTR HOSP QUEBEC CITY 10 QUEBEC CANADA OHIO STATE UNIV COLUMBUS OH 43210 USA TULANE UNIV SCH MED NEW ORLEANS LA 70118 USA VET ADM HOSP NEW ORLEANS LA 70112 USA

A series of D-amino acid-substituted analogs of the opiate peptide, methionine5-enkephalin, were synthesized by solid-phase methods and tested for their abilities to inhibit electrically-evoked contractions of mouse vas deferens and to complete with 3H enkephalin for opiate receptors on particulate fractions isolated from rat brain homogenates. [D-Ala2]-enkephalin and [D-Ala2]-enkephalin amide were the most potent peptides in both assay systems, being about 1000% active in the vas deferens bioassay and 120% and 150% active, respectively, in the stereospecific binding test relative to methionine5-enkephalin itself. In comparison, [D-Met5]-, [D-Tyr1]-, [D-Leu2]-, [D-Phe2]-, [D-Ala3]- and [D-Phe4]-enkephalin had no more than 10% activity. Stabilization of the .beta.-bend conformation of methionine5-enkephalin by the substitution of D-alanine in position 2 of the peptide chain may contribute to high activities of [D-Ala2]-analogs.

关键词：氨基酸类/分析生物学鉴定法甲硫氨酸寡肽类/化学合成寡肽类/药理学旋光度受体药物/代谢立体异构现象构效关系输精管/药物作用动物男(雄)性小鼠

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Ring constrained analogues of the orvinols: The furanomorphides.

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BIOORGANIC & MEDICINAL CHEMISTRY LETTERS 1999年第6期9卷 831-834页

作者： Husbands, SM Breeden, SW Grivas, K Lewis, JW Univ Bristol Sch Chem Bristol BS8 1TS Avon England

A series of furanomorphides were synthesised as ring-constrained analogues of buprenorphine and related orvinols. Evaluation in binding and functional assays has shown that the furanomorphides have reduced efficacy at... 详细信息

关键词：脑/药物作用丁丙诺啡/类似物和衍生物丁丙诺啡/药理学 CHO细胞仓鼠亚科呋喃类/化学合成半数抑制浓度动力学模型化学吗啡烷类/化学合成受体阿片样/药物作用输精管/药物作用动物豚鼠男(雄)性小鼠

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Further studies on nociceptin-related peptides: Discovery of a new chemical template with antagonist activity on the nociceptin receptor

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JOURNAL OF MEDICINAL CHEMISTRY 2000年第15期43卷 2805-2813页

作者： Guerrini, R Calo, G Bigoni, R Rizzi, A Varani, K Toth, G Gessi, S Hashiba, E Hashimoto, Y Lambert, DG Borea, PA Tomatis, R Salvadori, S Regoli, D Univ Ferrara Dept Expt & Clin Med Pharmacol Sect I-44100 Ferrara Italy Univ Ferrara Dept Pharmaceut Sci I-44100 Ferrara Italy Univ Ferrara Ctr Biotechnol I-44100 Ferrara Italy Biol Res Ctr Inst Biochem Isotope Lab H-6701 Szeged Hungary Univ Leicester Leicester Royal Infirm Dept Anaesthesia Leicester LE1 5WW Leics England

Three series of nociceptin (NC)-related peptides were synthesized and their abilities (i) to bind to the NC sites expressed in mouse forebrain membranes, (ii) to inhibit the electrically evoked contraction of the mouse vas deferens, and (iii) to inhibit forskolin-stimulated cAMP accumulation in Chinese hamster ovary cells expressing the human recombinant NC receptor (CHONCR) were investigated. The compounds of the first series (a series) have an ordinary Xaa(1)-Gly(2) bond, those of the second series (b series) have a Xaa(1)Psi(CH2-NH)Gly(2) pseudopeptide bond, and those of the third series (c series) have a peptoid (Nxaa(1)-Gly(2)) structure. The affinity values measured in the binding assay and in the two functional assays with the compounds of the three series showed high levels of correlation. Thus, (I) the compounds of the a series in which Phe(1) was substituted with Tyr, Cha, or Leu acted as potent NC receptor agonists;(II) the b series compounds behaved as NC receptor antagonists in the mouse vas deferens and as full agonists in CHONCR cells with different potencies depending on the first amino acid residue, [Phe(1)Psi(CH2-NH)Gly(2)]NC(1-17)NH2 and [Phe(1)Psi(CH2-NH)Gly(2)]NC(1-13)NH2 being the most potent compounds;(III) the compounds of the third series were all inactive both as agonists and as antagonists with the exception of [Nphe(1)]NC(1-17)NH2 and [Nphe(1)]NC(1-13)NH2, which behaved as NC receptor antagonists both in the isolated tissue and in CHONCR cells (pK(B) 6.1-6.4). In conclusion, this study demonstrates that chemical requirements for NC receptor agonists are different from those of antagonists. Moreover, modifications of the steric orientation of the aromatic residue Phe(1) in the NC sequence as obtained with the pseudopeptide bond between Phe(1) and Gly(2) or with the displacement of the benzyl side chain by one atom, as in Nphe(1), lead respectively to reduction or elimination of efficacy. Indeed, in contrast to [Phe(1)Psi(CH2-NH)Gly(2)]NC(1

关键词：脑/代谢 CHO细胞仓鼠亚科环AMP/代谢配体肌收缩/药物作用肌平滑/药物作用肌平滑/生理学麻醉品拮抗药阿片样肽类/化学肽类/化学合成肽类/化学肽类/代谢肽类/药理学类肽类蛋白质结合受体阿片样/激动剂重组蛋白质类/化学合成重组蛋白质类/化学重组蛋白质类/代谢重组蛋白质类/药理学构效关系输精管/药物作用输精管/生理学动物人类男(雄)性小鼠

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Synthesis, opioid receptor binding, and biological activities of naltrexone-derived pyrido- and pyrimidomorphinans

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JOURNAL OF MEDICINAL CHEMISTRY 1999年第18期42卷 3527-3538页

作者： Ananthan, S Kezar, HS Carter, RL Saini, SK Rice, KC Wells, JL Davis, P Xu, H Dersch, CM Bilsky, EJ Porreca, F Rothman, RB So Res Inst Dept Organ Chem Birmingham AL 35255 USA NIDA Clin Psychopharmacol Sect DIR Baltimore MD 21224 USA NIDDKD Med Chem Lab NIH Bethesda MD 20892 USA Univ No Colorado Dept Biol Sci Greeley CO 80639 USA Univ Arizona Coll Med Dept Pharmacol Tucson AZ 85724 USA

A series of pyrido- and pyrimidomorphinans (6a-h and 7a-g) were synthesized from naltrexone and evaluated for binding and biological activity at the opioid receptors. The unsubstituted pyridine 6a displayed high affinities at opioid delta, mu, and kappa receptors with Ki values of 0.78, 1.5, and 8.8 nM, respectively. Compound 6a was devoid of agonist activity in the mouse vas deferens (MVD) and guinea pig ileum (GPI) preparations but was found to display moderate to weak antagonist activity in the MVD and CPI with K-e values of 37 and 164 nM, respectively. The pyrimidomorphinans in general displayed lower binding potencies and 6 receptor binding selectivities than their pyridine counterparts. Incorporation of aryl groups as putative 6 address mimics on the pyrido- and pyrimidomorphinan framework gave ligands with significant differences in binding affinity and intrinsic activity. Attachment of a phenyl group at the 4'-position of 6a or the equivalent 6'-position of 7a led to dramatic reduction in binding potencies at all the three opioid receptors, indicating the existence of a somewhat similar steric constraint at the ligand binding sites of delta, mu, and kappa receptors. In contrast, the introduction of a phenyl group at the 6'-position of 6a did not cause any reduction in the binding affinity at the 6 receptor. In comparison to the unsubstituted pyridine 6a, the 5'-phenylpyridine 6c showed improvements in mu/delta and kappa/delta binding selectivity ratios as well as in the 6 antagonist potency in the MVD. Interestingly, introduction of a chlorine atom at the para position of the pendant 5'-phenyl group of 6c not only provided further improvements in 6 antagonist potency in the MVD but also shifted the intrinsic activity profile of sc from an antagonist to that of a mu agonist in the GPI. Compound Sd thus possesses the characteristics of a nonpeptide mu agonist/delta antagonist ligand with high affinity at the 6 receptor (Ki = 2.2 nM), high antagonist potency in

关键词：镇痛药/化学合成镇痛药/药理学结合竞争性脑/药物作用脑啡肽丙(2)-甲基苯丙(4)-甘(5)- 脑啡肽亮氨酸-2-丙氨酸/代谢脑啡肽类/代谢回肠/药物作用小鼠近交系吗啡烷类/化学合成吗啡烷类/药理学吗啡/药理学肌收缩/药物作用纳曲酮/化学蛋白质结合受体阿片样/代谢输精管/药物作用动物豚鼠男(雄)性小鼠大鼠

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BU48:: A novel buprenorphine analog that exhibits δ-opioid-mediated convulsions but not δ-opioid-mediated antinociception in mice

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JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS 2000年第3期294卷 1195-1200页

作者： Broom, DC Guo, L Coop, A Husbands, SM Lewis, JW Woods, JH Traynor, JR Univ Michigan Sch Med Dept Pharmacol Ann Arbor MI 48109 USA Univ Michigan Sch Med Dept Psychol Ann Arbor MI USA Univ Bristol Dept Chem Bristol Avon England Univ Loughborough Dept Chem Loughborough Leics England

N-Cyclopropylmethyl-[7 alpha,8 alpha,2',3']-cyclohexano-1'[S]-hydroxy-6,14-endo-ethenotetrahydronororipavine (BU48) is a novel, ring-constrained analog of buprenorphine. In vivo, BU48 (0.1-10 mg/kg s.c.) produced brief, nonlethal convulsions in mice followed by brief Straub tail and a short period of catalepsy characteristic of BW373U86 and other nonpeptidic delta-receptor agonists. BU48-induced convulsions were sensitive to antagonism by naltrindole (10 mg/kg s.c.) and were also prevented by administration of the putative delta(1) antagonist 7-benzylidenenaltrexone and the putative delta(2) antagonist naltriben, with the latter being more potent. In the abdominal stretch assay in the mouse, only low-efficacy antinociceptive activity of BU48 (0.1-10 mg/kg) was seen. This was reversed by the kappa-opioid antagonist norbinaltorphimine (32 mg/kg s.c.) but not by the delta-opioid antagonist naltrindole (10 mg/kg s.c.). BU48 (10 mg/kg s.c.) acted as a delta-antagonist in this assay. In mouse brain homogenates, BU48 had high (nanomolar) binding affinity for all three opioid receptors in the order mu > delta = kappa. In vitro, the compound acted as a potent (EC50 = 1.4 nM) kappa-opioid agonist in the guinea pig ileum and a potent (EC50 = 0.2 nM) delta-opioid agonist in the mouse vas deferens but showed partial agonist activity at the rat cloned delta-opioid (40%) and human cloned kappa-opioid (59%) receptors with very low efficacy at the rat cloned mu-opioid receptor (10%);findings consistent with its in vivo profile. BU48 is the first described compound that produces delta-opioid-mediated convulsions without any evidence of delta-opioid-mediated antinociception and will be a useful tool in investigations of the delta-opioid receptor.

关键词：镇痛药阿片类/药理学苯甲酰胺类/药理学脑/代谢丁丙诺啡/类似物和衍生物丁丙诺啡/药理学惊厥药/药理学电刺激鸟苷5'-O-(3-硫代三磷酸)/代谢回肠/药物作用回肠/生理学肌收缩/药物作用肌平滑/药物作用肌平滑/生理学纳曲酮/类似物和衍生物纳曲酮/药理学麻醉品拮抗药/药理学疼痛测定哌嗪类/药理学放射配体测定受体阿片样 δ/激动剂受体阿片样 δ/拮抗剂和抑制剂受体阿片样 δ/生理学受体阿片样 κ/激动剂受体阿片样 κ/拮抗剂和抑制剂发作/化学诱导输精管/药物作用输精管/生理学动物豚鼠男(雄)性小鼠大鼠

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