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71.
72.
B A Vandekerckhove J F Krowka J M McCune J E de Vries H Spits M G Roncarolo 《Journal of immunology (Baltimore, Md. : 1950)》1991,146(12):4173-4179
Severe combined immunodeficiency (SCID) mice can be transplanted successfully with human fetal liver and thymus (SCID-hu mice). Precursor cells derived from the fetal liver differentiate in the thymus and migrate into the blood as mature T cells. In the present paper, the peripheral T cell compartment of such mice was studied. Peripheral WBC were activated by PHA and cultured in the presence of irradiated human feeder cells. The resultant cell population consisted exclusively of human CD1- CD2+ CD3+ CD7+ T lymphocytes; up to 4% of the T cells expressed the TCR gamma delta, whereas 95 to 100% were TCR alpha beta +. The CD4bright (42 to 66%) and CD8bright (30 to 54%) populations coexpressed variable but low levels of CD8 and CD4, respectively. The T cell cultures from the SCID-hu mice did not display reactivity towards the autologous human EBV-transformed B cell lines (B-LCL). On the other hand, these human T cells proliferated and were cytotoxic against allogeneic human B-LCL. T cell clones were established from cultured SCID-hu T cells. All T cell clones were TCR alpha beta + CD3+ CD2+; 61% of the clones were CD4+ CD8-, 27% were CD8+ CD4-, 11% were CD8+ CD4lo, and 2% were CD4+ CD8lo. None of these clones recognized the autologous B-LCL established from the fetal human donor. Fourteen of 100 T cell clones had specific alloreactivity, as tested on a panel of five B-LCL. Of these 14, two CD8+ CD4lo and two CD8+ CD4- clones were cytotoxic and did not proliferate in response to specific stimulator cells. Furthermore, two CD4+ CD8lo and eight CD4+ CD8- clones proliferated specifically in response to alloantigens. In conclusion, the peripheral human T cells of SCID-hu animals are functional and their TCR repertoire is polyclonal, alloreactive, and devoid of self-reactive cells. Therefore, the SCID-hu mouse can be a suitable model for the study of alloreactivity and allotolerance in vivo, as well as for the study of negative selection in the human thymus. 相似文献
73.
To investigate the relationships among the three main groups of extant neopterygian fishes--Amiidae, Lepisosteidae, and Teleostei--we sequenced fragments of three mitochondrial genes from 12 different actinopterygian fishes and translated the nucleotide sequences into amino acid sequences. When all three regions are considered together, Amiidae clusters with Lepisosteidae in the most parsimonious cladograms, but other clades, such as Neopterygii and Teleostei, that are well supported by morphological evidence fail to emerge as monophyletic. When the cytochrome b sequences are analyzed together with previously published sequences for other taxa, the majority-rule consensus tree is consistent with the monophyly of Teleostei and Neopterygii and marginally supports the Amiidae + Lepisosteidae clade. In either analysis, when Neopterygii and Teleostei are constrained to monophyly, all the most-parsimonious cladograms support the Amiidae + Lepisosteidae topology. Where molecules and morphology disagree, provisional morphology-based constraints on the analysis of molecular data offer a practical means of integrating the two types of data. 相似文献
74.
Kenneth C. Kuo Charles W. Gehrke Roy A. McCune 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1978,145(3):383-392
A rapid, precise, and accurate chromatographic method for the determination of pseudo-uridine (ψ) in urine by high-performance liquid chromatography (HPLC) has been developed. The ribonucleosides were first isolated with an affinity gel containing immobilized phenylboronic acid. The response for ψ was linear well above and below the range necessary to determine urinary ψ. Good precision was obtained for both matrix-dependent and matrix-independent samples. Supporting experimental data are presented on precision, recovery, chromatographic methods, sample cleanup and application to the analysis of urine samples from normal males and females, and patients with advanced colon cancer. In a comparison of 40 normals with 10 colon cancer patients, 9 of the 10 patients had a ψ: creatinine (Cr) ratio greater than
+ 2σ for the normal population. This HPLC method is now being used extensively in our laboratory as a routine method for determination of ψ in urine from patients with various types of cancer and in chemotherapy response studies. Data are presented on the dynamics of ψ excretion by normal males and females. When the excretion of ψ was normalized with the excretion of creatinine, it was noted that samples collected at random have the same ψ: Cr ratio value as for the 24-h total collection, thus, allowing the use of random samples. The constancy of the ψ: Cr ratio implies that RNA turnover is constant and ψ excretion is independent of diet. Base values are presented for the ψ: Cr 相似文献
75.
Rapid-high, syncytium-inducing isolates of human immunodeficiency virus type 1 induce cytopathicity in the human thymus of the SCID-hu mouse. 总被引:13,自引:9,他引:4
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H Kaneshima L Su M L Bonyhadi R I Connor D D Ho J M McCune 《Journal of virology》1994,68(12):8188-8192
Clinical deterioration in human immunodeficiency virus type 1 (HIV-1) disease is associated with an increased viral burden in the peripheral blood and a loss of circulating CD4+ T cells. HIV-1 isolates obtained prior to this stage of disease often have a "slow-low," non-syncytium-inducing (NSI) phenotype, whereas those obtained afterwards are often characterized as "rapid-high" and syncytium inducing (SI). Paired NSI and SI isolates from two different patients were inoculated into the human thymus implants of SCID-hu mice. The two slow-low, NSI isolates replicated to minimal levels in the grafts and did not induce thymocyte depletion. In contrast, the two SI isolates from the same patients showed high levels of viral replication and induced a marked degree of thymocyte depletion, accompanied by evidence of programmed cell death. These observations reveal a correlation between the replicative and cytopathic patterns of HIV-1 isolates in vitro and in the SCID-hu mouse in vivo and provide direct evidence that the biological phenotype of HIV-1 switch may be a causal and not a derivative correlate of HIV-1 disease progression. 相似文献
76.
77.
H S Earp J R Hepler L A Petch A Miller A R Berry J Harris V W Raymond B K McCune L W Lee J W Grisham 《The Journal of biological chemistry》1988,263(27):13868-13874
Epidermal growth factor (EGF) stimulated the rapid accumulation of inositol trisphosphate in WB cells, a continuous line of rat hepatic epithelial cells. Since we previously had shown that EGF stimulates EGF receptor synthesis in these cells, we tested whether hormones that stimulate PtdIns(4,5)P2 hydrolysis would increase EGF receptor protein synthesis and mRNA levels. Epinephrine, angiotensin II, and [Arg8]vasopressin activate phospholipase C in WB cells as evidenced by the accumulation of the inositol phosphates, inositol monophosphate, inositol bisphosphate, and inositol trisphosphate. A 3-4-h treatment with each hormone also increased the rate of EGF receptor protein synthesis by 3-6-fold as assessed by immunoprecipitation of EGF receptor from [35S]methionine-labeled cells. Northern blot analyses of WB cell EGF receptor mRNA levels revealed that agents linked to the phosphoinositide signaling system increased receptor mRNA content within 1-2 h. A maximal increase of 3-7-fold was observed after a 3-h exposure to EGF and hormones. The phorbol ester, 12-O-tetradecanoylphorbol 13-acetate (TPA), which activates protein kinase C also stimulated EGF receptor synthesis. Pretreatment of WB cells for 18 h with high concentrations of TPA "down-regulated" protein kinase C and blocked TPA-directed EGF receptor mRNA synthesis. In contrast, the effect of EGF on EGF receptor mRNA levels was not significantly decreased by TPA pretreatment. Epinephrine-induced increases in EGF receptor mRNA were reduced from 4- to 2-fold. Similarly, 18 h TPA pretreatment abolished the effect of TPA on EGF receptor protein synthesis but did not affect EGF-dependent EGF receptor protein synthesis. The 18-h TPA pretreatment diminished by 30-50% the induction of receptor protein synthesis by epinephrine or angiotensin II. We conclude that in WB cells EGF receptor synthesis can be regulated by EGF and other hormones that stimulate PtdIns(4,5)P2 hydrolysis. In these cells, EGF receptor synthesis appears to be regulated by several mechanism: one pathway is dependent upon EGF receptor activation and can operate independently of protein kinase C activation; another pathway is correlated with PtdIns(4,5)P2 hydrolysis and is dependent, at least in part, upon protein kinase C activation. 相似文献
78.
Bruce McCune 《植被学杂志》1992,3(1):27-34
Abstract. A method is given for measuring two components of error (rate and direction) in predictions of compositional change through time. Observed compositional change between two times can be represented as a vector between two points in multidimensional species space. The point at the tail of this vector is the species composition at one particular time. A vector of predicted compositional change will diverge from the vector of observed change to some degree. The error in the predicted rate of change is measured by the difference between the lengths of the two vectors. The error in the predicted direction of change is measured by the angle between the vectors. The cosine of this angle is a non-standardized correlation coefficient (rn) between the predicted and observed species compositions. The quantity 1 - rn2 measures the error in direction of the predicted dynamics without being influenced by the overall rate of change. These measures in Euclidean space have useful counterparts in city-block space. The method is illustrated by comparing actual long-term changes in Midwestern old-growth forests with the changes predicted by a growth and yield model, TWIGS. 相似文献
79.
W. S. McCune 《BMJ (Clinical research ed.)》1908,2(2490):784-785
80.
Timothy J. Nice David W. Strong Broc T. McCune Calvin S. Pohl Herbert W. Virgin 《Journal of virology》2013,87(1):327-334
Human norovirus (HuNoV) is the major cause of acute nonbacterial gastroenteritis worldwide but has no clear animal reservoir. HuNoV can persist after the resolution of symptoms, and this persistence may be essential for viral maintenance within the population. Many strains of the related murine norovirus (MNV) also persist, providing a tractable animal model for studying norovirus (NoV) persistence. We have used recombinant cDNA clones of representative persistent (CR6) and nonpersistent (CW3) strains to identify a domain within the nonstructural gene NS1/2 that is necessary and sufficient for persistence. Furthermore, we found that a single change of aspartic acid to glutamic acid in CW3 NS1/2 was sufficient for persistence. This same conservative change also caused increased growth of CW3 in the proximal colon, which we found to be a major tissue reservoir of MNV persistence, suggesting that NS1/2 determines viral tropism that is necessary for persistence. These findings represent the first identified function for NoV NS1/2 during infection and establish a novel model system for the study of enteric viral persistence. 相似文献