全文获取类型
收费全文 | 53790篇 |
免费 | 5680篇 |
国内免费 | 324篇 |
出版年
2022年 | 402篇 |
2021年 | 780篇 |
2020年 | 569篇 |
2019年 | 746篇 |
2018年 | 970篇 |
2017年 | 835篇 |
2016年 | 1273篇 |
2015年 | 1777篇 |
2014年 | 2000篇 |
2013年 | 2573篇 |
2012年 | 2927篇 |
2011年 | 2847篇 |
2010年 | 1868篇 |
2009年 | 1624篇 |
2008年 | 2269篇 |
2007年 | 2123篇 |
2006年 | 2071篇 |
2005年 | 1839篇 |
2004年 | 1837篇 |
2003年 | 1729篇 |
2002年 | 1744篇 |
2001年 | 3283篇 |
2000年 | 3022篇 |
1999年 | 2317篇 |
1998年 | 853篇 |
1997年 | 871篇 |
1996年 | 707篇 |
1995年 | 677篇 |
1994年 | 518篇 |
1993年 | 519篇 |
1992年 | 1272篇 |
1991年 | 1118篇 |
1990年 | 985篇 |
1989年 | 830篇 |
1988年 | 661篇 |
1987年 | 588篇 |
1986年 | 519篇 |
1985年 | 523篇 |
1984年 | 440篇 |
1983年 | 382篇 |
1982年 | 280篇 |
1981年 | 281篇 |
1980年 | 242篇 |
1979年 | 311篇 |
1978年 | 277篇 |
1977年 | 294篇 |
1976年 | 284篇 |
1975年 | 273篇 |
1974年 | 273篇 |
1973年 | 268篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
981.
The antisense therapeutic strategy makes the assumption that sequence-specific hybridization of an oligonucleotide to its target can take place in living cells. The present work provides a new method for the detection of intracellular RNA molecules using in situ hybridization on living cells. The first step consisted in designing nonperturbant conditions for cell permeabilization using streptolysin O. In a second step, intracellular hybridization specificity was evaluated by incorporating various types of fluorescently labeled nucleic acid probes (plasmids, oligonucleotides). Due to its high expression level, the 28S ribosomal RNA was retained as a model. Results showed that: (1) no significant cell death was observed after permeabilization; (2) on living cells, 28S RNA specific probes provided bright nucleoli and low cytoplasmic signal; (3) control probes did not lead to significant fluorescent staining; and (4) comparison of signals obtained on living and fixed cells showed a colocalization of observed fluorescence. These results indicate the feasibility of specific hybridization of labeled nucleic acid probes under living conditions, after a simple and efficient permeabilization step. This new detection method is of interest for investigating the dynamics of distribution of various gene products in living cells, under normal or pathological conditions.Abbreviations PI
propidium iodide
- SLO
streptolysin O 相似文献
982.
The complete mitochondrial DNA (mtDNA) molecule of the gorilla was
sequenced. The entire sequence, 16,412 nucleotides, was determined by
analysis of natural (not polymerase chain reaction) restriction fragments
covering the whole molecule. The sequence was established from one
individual and thus nonchimeric. After comparison with the COII gene of
gorilla specimens with known geographical origin, the sequence was
identified as characteristic of the Western lowland gorilla, Gorilla
gorilla gorilla. With the exception of the NADH2 gene, all genes have a
methionine start codon. The inferred start codon of NADH2 is ATT
(isoleucine). The COIII, NASDH4, and cytochrome b genes are not terminated
by a stop codon triplet, and the COI gene is probably terminated by an AAA
triplet rather than by a regular stop codon. The great majority of genic
sequences (rRNAs, peptide-coding genes, tRNAs) of the complete mtDNAs of
Gorilla, Pan, and Homo show a greater similarity between Pan and Homo than
between either of these genera to Gorilla. The analysis of the
peptide-coding genes suggest that relative to comparison between Homo and
Pan a certain degree of transition saturation has taken place in codon
position 3 in comparisons between Gorilla to either Homo or Pan.
相似文献
983.
Two major problems associated with biofouling studies are the lack of broad scale testing and failure to use consistent standards among different assays or studies. To address these issues the activity of two biologically active natural products, elatol and deschloroelatol, isolated from the marine red alga Laurencia rigida, and three commonly used biocides, Nopcocide N-96?, Irgarol 1051? and Sea-Nine 211?, was compared, in a broad spectrum of bioassays. The activity of the different compounds varied substantially among different bioassay tests. Elatol and deschloroelatol had a narrow range of activity with strongest effects against invertebrate larvae. Both compounds were highly toxic. However, neither compound had strong activity against marine bacteria or the common epiphyte Ulva lactuca. Irgarol 1051 also had a narrow range of activity, only affecting algal settlement strongly. Nopcocide N-96 and Sea-Nine 211 had moderate to strong activity across the spectrum of bioassays, viz. growth of marine bacteria (Vibrio fischeri, Serratia sp.), inhibition of settlement of macroalgae (Ulva lactuca), toxicity (Balanus amphitrite), and inhibition of settlement of invertebrate larvae (Balanus amphitrite, Bugula neritina). Based on the results it is proposed that Sea-Nine 211, because of its broad spectrum activity, be used as a standard for comparative assessments of the antifouling activity of marine natural products and analogues. 相似文献
984.
Transfer factor in chronic mucocutaneous candidiasis 总被引:3,自引:0,他引:3
Fifteen patients suffering from chronic mucocutaneous candidiasis were treated with an in vitro produced TF specific for Candida
albicans antigens and/or with TF extracted from pooled buffy coats of blood donors. CMI of the patients was assessed using
the LMT and the LST in presence of candidine. The aim of the study was the clinical evaluation of TF treatment and the incidence
of positive tests before, during, and after therapy. Immunological data were matched using the Chi square test. 87 LMT were
performed for each antigen dose and at the dilution of 1/50, 58.9% (33/56) tests were positive during non-treatment or non-specific
TF treatment. On the contrary 83.9% (26/31) were positive during specific TF treatment (P<0.05). In the LST, a significant
decrease of thymidine uptake in the control cultures in presence of autologous or AB serum was observed when patients were
matched according to non-treatment, and both non specific (P<0.05) and specific TF treatment (P<0.01). Only during specific
TF treatment was a significant increase of reactivity against the Candida antigen at the highest concentration noticed, when
compared with the period of non specific treatment (P<0.01). Clinical observations were encouraging: all but one patient experienced
significant improvement during treatment with specific TF. These data confirm that orally administered specific TF, extracted
from induced lymphoblastoid cell-lines, increases the incidence of reactivity against Candida antigens in the LMT. LST reactivity
appeared not significantly increased with respect to the periods of non treatment, but was significantly increased when it
was compared to the non-specific TF treatment periods. At the same time, a clinical improvement was noticed. 相似文献
985.
A preliminary report on the use of transfer factor for treating stage D3 hormone-unresponsive metastatic prostate cancer 总被引:4,自引:0,他引:4
Dr. Giancarlo Pizza Caterina De Vinci Diego Cuzzocrea Domenico Menniti Ernesto Aiello Paolo Maver Giuseppe Corrado Piero Romagnoli Ennio Dragoni Giuseppe LoConte Umberto Riolo Aldopaolo Palareti Paolo Zucchelli Vittorio Fornarola Dimitri Viza 《Biotherapy》1996,9(1-3):123-132
As conventional treatments are unsuccessful, the survival rate of stage D3 prostate cancer patients is poor. Reports have
suggested the existence of humoral and cell-mediated immunity (CMI) against prostate cancer tumour-associated antigens (TAA).
These observations prompted us to treat stage D3 prostate cancer patients with an in vitro produced transfer factor (TF) able
to transfer, in vitro and in vivo, CMI against bladder and prostate TAA. Fifty patients entered this study and received one
intramuscular injection of 2–5 units of specific TF monthly. Follow-up, ranging from 1 to 9 years, showed that complete remission
was achieved in 2 patients, partial remission in 6, and no progression of metastatic disease in 14. The median survival was
126 weeks, higher than the survival rates reported in the literature for patients of the same stage. 相似文献
986.
Prof. Renato Meduri Emilio Campos Lucia Scorolli Caterina De Vinci Giancarlo Pizza Dimitri Viza 《Biotherapy》1996,9(1-3):61-66
Recurrent ocular herpes is an insoluble problem for the clinician. As cellular immunity plays an important role in controlling
herpes relapses, and other studies have shown the efficacy of HSV-specific transfer factor (TF) for the treatment of herpes
patients, an open clinical trial was undertaken in 134 patients (71 keratitis, 29 kerato-uveitis, 34 uveitis) suffering from
recurrent ocular herpetic infections. The mean duration of the treatment was 358 days, and the entire follow-up period 189121
before, and 64062 days after TF treatment. The cell-mediated immune response to the viral antigens, evaluated by the lymphocyte
stimulation test (LST) and the leucocyte migration test (LMT) (P<0.001), was significantly increased by the TF treatment.
The total number of relapses was decreased significantly during/after TF treatment, dropping from 832 before, to 89 after
treatment, whereas the cumulative relapse index (RI) dropped, during the same period, from 13.2 to 4.17 (P<0.0001). No side
effects were observed. It is concluded that patients with relapsing ocular herpes can benefit from treatment with HSV-specific
TF. 相似文献
987.
Hybridomas were cultured under steady-state conditions in a series of two continuous stirred-tank reactors (CSTRs), using a serum-free medium. The substrate not completely converted in the first CSTR, was transported with the cells to the second one and very low growth rates, high death rates, and lysis of viable cells were observed in this second CSTR. These conditions are hardly accessible in a single vessel, because such experiments would be extremely time-consuming and unstable due to a low viability. In contrast to what is often observed in literature, kinetic parameters could thus be derived without the neccessity for extrapolation to lower growth rates. Good agreement with literature averages for other hybridomas was found. Furthermore, showing that the reactor series is a valuable research tool for kinetic studies under extreme conditions, the possibility to observe cell death under stable and defined steady-state conditions offers interesting opportunities to investigate apoptosis and necrosis. Additionally, a model was developed that describes hybridoma growth and monoclonal antibody production in the bioreactor cascade on the basis of glutamine metabolism. Good agreement between the model and the experiments was found.Abbreviation MAb
Monoclonal antibody
Nomenclature
C
AConcentration of any (mol m-3) component A
D Dilution rate (s-1)
K
dDeath-rate constant (mol m-3)
K
lLysis-rate constant (mol m-3)
K
sMonod constant (mol m-3)
m Maintenance coefficient (mol cell-1 s-1)
q Specific consumption (mol cell-1 s-1) or production rate
t Time (s)
X Cell concentration (cell m-3)
Y Yield coefficient (cell mol-1)
Greek symbols
d
Specific death rate (s-1)
l
Specific lysis rate (s-1) of viable cells
net
Net specific growth (s-1) rate
true
True specific growth (s-1) rate 相似文献
988.
An efficient impedance method was developed for rapid evaluation of cosmetic preservatives. The method used decimal reduction time or D-value to assess preservative efficacies. The D-value, which was calculated from the plot of Log CFU ml–1 versus time by linear regression analysis, could be obtained within 48 h. Thus, the time required for the challenge test was reduced from 4–8 weeks with the standard procedures (eg US Pharmacopeia), to 2 days with the current method. A calibration curve (r=-0.95) was established by plotting the Log CFU ml–1 versus capacitance detection time (DT) of 108 samples. With the calibration, CFU can be estimated directly from the impedance test without plating. Two commercial biocides and several other chemicals were evaluated in a shampoo by the impedance procedure againstPseudomonas aeruginosa. The D-values obtained from the impedance test were not significantly different from those produced by the conventional plate count method. The technique was found to be particularly useful when screening a large number of compounds to find novel preservatives and synergistic preservative combinations. 相似文献
989.
Cornelia Herschbach L. J. De Kok H. Rennenberg 《Plant biology (Stuttgart, Germany)》1995,108(1):41-46
Spinach plants (Spinacea oleracea L. cv. Estivato) were grown on nutrient solutions under deficient, normal and excess sulfate supply. In both young and mature plants net uptake of sulfate and its transport to the shoot increased with increasing sulfate supply, but both processes proceeded at a higher rate in young as compared to mature plants. The relative sulfate transport, i.e. the relative amount of the sulfate taken up that is transported to the shoot, decreased with increasing sulfate supply. Apparently, net uptake of sulfate is not strictly controlled by the sulfur demand of the shoot, but xylem loading appears to counteract excess transport of sulfate to the shoot. Fumigation with H2S or SO2 reduced net uptake of sulfate by the roots in sulfur-deficient plants and absolute as well as relative sulfate transport to the shoot independent of the three sulfate levels supplied to the plant. At the same time thiol contents of the shoot and the root were enhanced by fumigation with H2S and SO2. These findings are consistent with the idea that thiols produced in the leaves can mediate demand-driven control of sulfate uptake by the roots and its transport to the shoot. 相似文献