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991.
AS Kolhekar HT Keutmann RE Mains AS Quon BA Eipper 《Canadian Metallurgical Quarterly》1997,36(36):10901-10909
Peptidylglycine alpha-hydroxylating monooxygenase (PHM) is a copper, ascorbate, and molecular oxygen dependent enzyme that catalyzes the first step leading to the C-terminal amidation of glycine-extended peptides. The catalytic core of PHM (PHMcc), refined to residues 42-356 of the PHM protein, was expressed at high levels in CHO (DG44) (dhfr-) cells. PHMcc has 10 cysteine residues involved in 5 disulfide linkages. Endoprotease Lys-C digestion of purified PHMcc under nonreducing conditions cleaved the protein at Lys219, indicating that the protein consists of separable N- and C-terminal domains with internal disulfide linkages, that are connected by an exposed linker region. Disulfide-linked peptides generated by sequential CNBr and pepsin treatment of radiolabeled PHMcc were separated by reverse phase HPLC and identified by Edman degradation. Three disulfide linkages occur in the N-terminal domain (Cys47-Cys186, Cys81-Cys126, and Cys114-Cys131), along with three of the His residues critical to catalytic activity (His107, His108, and His172). Two disulfide linkages (Cys227-Cys334 and Cys293-Cys315) occur in the C-terminal domain, along with the remaining two essential His residues (His242, His244) and Met314, thought to be essential in binding one of the two nonequivalent copper atoms. Substitution of Tyr79 or Tyr318 with Phe increased the Km of PHM for its peptidylglycine substrate without affecting the Vmax. Replacement of Glu313 with Asp increased the Km 8-fold and decreased the kcat 7-fold, again identifying this region of the C-terminal domain as critical to catalytic activity. Taking into account information on the copper ligands in PHM, we propose a two-domain model with a copper site in each domain that allows spatial proximity between previously described copper ligands and residues identified as catalytically important. 相似文献
992.
HH Yap AS Chong CR Adanan NL Chong B Rohaizat YA Malik SY Lim 《Canadian Metallurgical Quarterly》1997,13(4):384-388
To study the effects of percutaneous ethanol injection therapy on livers, we investigated necrotic changes after ethanol injection and fibrotic changes during the repair process in cirrhotic livers in comparison with normal livers. Male rats were treated with oral doses of 3'-methyl-4-dimethylaminoazobenzene and thioacetamide to produce liver cirrhosis. Both control animals and cirrhotic animals were injected with 0.2 ml of absolute ethanol into livers. Histological samples were cut serially, and the maximum areas of necrosis and fibrosis were measured until 28 days after the injection. Although the maximum area of necrosis was not different between cirrhotic livers and control livers, the average fibrotic ratio [(maximum fibrotic area/maximum necrotic area + maximum fibrotic area) x 100] was 64% in control livers (n = 9) and 40% in cirrhotic livers (n = 9; p < 0.05). The fibrotic repair process after ethanol injection seems to be impaired in cirrhotic livers as compared with normal livers. 相似文献
993.
994.
OBJECTIVE: The present investigation was undertaken to determine the incidence of alcohol, cocaine, and marijuana use among pregnant patients who suffered any type of physical trauma and to determine if the combination of substance use and physical trauma in pregnancy has extended perinatal implications. METHODS: In this single institution, prospective patient series undertaken over 18 months, all pregnant patients who suffered any type of physical trauma were considered for study. RESULTS: Among 6828 live births over an 18-month period, there were 157 (2.3%) patients who reported physical trauma (insignificant = 153, minor = 2, severe = 2). Consent to screen for the presence of alcohol, marijuana, and cocaine was obtained from 85 (54%) patients with refusal by the remainder. Screening was positive in 11% for one or more substances. The perinatal morbidity and mortality for the 153 patients with insignificant trauma was 50:1000 and 20:1000, respectively, all associated with preterm labor and delivery which occurred in 21% of the patients. None of the four patients with minor or severe trauma either had a positive drug/alcohol screen or any perinatal morbidity or mortality. No maternal mortality occurred. CONCLUSION: An 11% incidence of positive drug screens was encountered in consenting pregnant trauma victims and a 21% incidence of preterm birth was encountered even in this patient population with so-called insignificant trauma. 相似文献
995.
GR Kershaw C Berger C McQuain AS al-Homsi G Pihan PJ Quesenberry BA Woda H Knecht 《Canadian Metallurgical Quarterly》1997,64(7):1079-1081
BACKGROUND: Posttransplant lymphoproliferative disorders are generally associated with Epstein-Barr virus (EBV) and are of B cell origin. We report the case of a B-immunoblastic lymphoma that developed in a pretransplantation EBV-seronegative woman 4 months after kidney transplant from her HLA-haploidentical brother. The patient successfully underwent immunotoxin therapy for lymphoma and has been in remission for 36 months. METHODS: Latent EBV genomes were identified by polymerase chain reaction, and the purified amplification products were directly sequenced with [35S]dATP. RESULTS: Molecular analysis of the latent membrane protein (LMP)1 oncogene of EBV, which was expressed in most tumor cells, revealed a 30-base pair deletion. No wild-type LMP1 sequences were found. Analysis of peripheral blood mononuclear cells from the EBV-seropositive donor showed the presence of both the LMP1 deletion variant and the wild-type sequence. The LMP1 deletion variant and the wild-type sequence were also identified within peripheral blood mononuclear cells of the EBV-seroconverted kidney recipient 20 months after lymphoma therapy. CONCLUSION: This pattern is consistent with a natural growth advantage of B cells expressing the LMP1 deletion variant in the immunocompromised host. 相似文献
996.
Undifferentiated glandular stomach tissue fragments from 16.5-day fetal rats were transplanted under the kidney capsule of syngeneic adult rats, and the proliferation, differentiation and morphogenesis of the transplanted tissues were investigated. Gastric epithelial cells began to invaginate 3-4 days after the transplantation and immature glands were formed after 1 week. During the period, there was a gradual increase in the expression of pepsinogen and cathepsin E, markers of cytodifferentiation of the stomach epithelia, both at protein and mRNA levels. Cathepsin E was weakly expressed in undifferentiated gastric epithelial cells at 16.5 days of gestation, and a higher level of the expression was observed in differentiated epithelia of the transplants. In contrast, the pepsinogen-producing cells first appeared around days 3-4 after transplantation and gradually increased in number to about 30% of the epithelial cells and became localized at the bottom of the gland. During the period of the experiment up to 1 month, the pepsinogen-producing cells were all positive for class III mucin and cathepsin E, indicating the immature character of these cells. In addition, no parietal cells were observed. When the tissue fragments were transplanted into adrenalectomized animals, the epithelial differentiation and morphogenesis was suppressed, but its proliferation was enhanced. The observed changes were reversed by hydrocortisone replacement. These results suggest that the development of the 16.5-day fetal stomach is regulated intrinsically to a certain extent by the genetic program of the cells involved and various gastric functions develop in the absence of luminal stimulation, stage-specific systemic hormonal change, neuronal regulation or other systemic influences, and that glucocorticoids modulate the developmental program of the fetal stomach tissues. 相似文献
997.
A Kafatos A Diacatou G Voukiklaris N Nikolakakis J Vlachonikolis D Kounali G Mamalakis AS Dontas 《Canadian Metallurgical Quarterly》1997,65(6):1882-1886
A follow-up study was conducted to identify the heart disease risk-factor status and dietary changes of surviving elderly subjects in Crete who took part in the Seven Countries Study in 1960. In 1991, data were obtained from 245 of the 686 original male participants (169 of the original 40-49-y age group and 76 men 50-59 y age group). In 1991, the men were 70-79 and 80-89 y old. There was a significant (11.5%) increase in serum total cholesterol concentrations between 1960 and 1991. Body mass index and systolic and diastolic blood pressures also increased significantly, and all age groups were characterized by central obesity. A representative subsample of 21 men took part in a 3-d weighed food record study. Dietary data indicated increases in the intake of saturated fat and decreases in monounsaturated fat over the 30-y period. Comparison with a 1962 representative Cretan sample indicated a significantly increased concentration of adipose palmitic acid (16:0) in our surviving sample. The observed changes occurred during a period when many developed countries were observing a decline in most heart disease risk factors. 相似文献
998.
999.
Rouleaux are formed by the aggregation of red blood cells in the presence of macromolecules that bridge the membranes of adherent erythrocytes. We compute the size and degree of branching of rouleaux for macroscopic systems in thermal equilibrium in the absence of fluid flow. Using techniques from statistical mechanics, analytical expressions are derived for (a) the average number of rouleaux consisting of n cells and having m branch points; (b) the average number of cells per rouleau; (c) the average number of branch points per rouleau; and (d) the number of rouleaux with n cells, n = 1, 2, ..., in a system containing a total of N cells. We also present the results of numerical evaluations to establish the validity of asymptotic expressions that simplify our formal analytic results. 相似文献
1000.
AS Pudenko IS Kudryavtsev OV Zatsepina Chentsov YuS 《Canadian Metallurgical Quarterly》1997,11(4):449-461
The literature has been reviewed to evaluate the technique and clinical value of radionuclide measurements of microvascular permeability and oedema formation in the lungs. Methodology, modelling and interpretation vary widely among studies. Nevertheless, most studies agree on the fact that the measurement of permeability via pulmonary radioactivity measurements of intravenously injected radiolabelled proteins versus that in the blood pool, the so-called pulmonary protein transport rate (PTR), can assist the clinician in discriminating between permeability oedema of the lungs associated with the adult respiratory distress syndrome (ARDS) and oedema caused by an increased filtration pressure, for instance in the course of cardiac disease, i.e. pressure-induced pulmonary oedema. Some of the techniques used to measure PTR are also able to detect subclinical forms of lung microvascular injury not yet complicated by permeability oedema. This may occur after cardiopulmonary bypass and major vascular surgery, for instance. By paralleling the clinical severity and course of the ARDS, the PTR method may also serve as a tool to evaluate new therapies for the syndrome. Taken together, the currently available radionuclide methods, which are applicable at the bedside in the intensive care unit, may provide a gold standard for detecting minor and major forms of acute microvascular lung injury, and for evaluating the severity, course and response to treatment. 相似文献