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1.
The mechanism of bicarbonate transport across the peritubular cell membrane was investigated in rat kidney proximal tubules in situ by measuring cell pH and cell Na+ activity in response to sudden reduction of peritubular Na+ and/or HCO 3 . The following observations were made: 1. sudden peritubular reduction of either ion concentration produced the same transient depolarizing potential response; 2. bicarbonate efflux in response to peritubular reduction of bicarbonate was accompanied by sodium efflux; 3. sodium efflux in response to peritubular sodium removal was accompanied by cell acidification indicating bicarbonate efflux; 4. all aforementioned phenomena were inhibited by SITS (10–3 mol/l) except for a small SITS-independent sodium efflux and depolarization which occurred in response to peritubular sodium removal and was not accompanied by cell pH changes; 5. bicarbonate efflux and accompanying potential changes in response to reduction of peritubular bicarbonate virtually vanished in sodium-free solutions. From these observations we conclude that bicarbonate efflux proceeds as rheogenic sodium-bicarbonate cotransport with a stoichiometry of bicarbonate to sodium greater than 1. The question which of the charged species of the bicarbonate buffer system moves cannot yet be decided. Attempts to determine the stoichiometry from the SITS-inhibitable initial cell depolarization and from the SITS-inhibitable initial fluxes suggest a stoichiometry of 3 HCO 3 : 1 Na+. In addition to sodium-dependent bicarbonate flux, evidence was obtained for a sodium-independent transport system of acids or bases which is able to regulate cell pH even in sodium-free solutions.  相似文献   
2.
The proton dependence of the transport of three labelled, hydrolysis-resistant synthetic dipeptides carrying a net charge of –1, 0 or +1 has been investigated in a brush border membrane vesicle preparation obtained from rat renal cortex. Cross-inhibition studies are consistent with the transport of all peptides studied being through a single system. The extent and time course of uptake in response to an inwardly directed electrochemical gradient of protons differed for each peptide. For the cationic peptide D-Phe-L-Lys this gradient did not stimulate the initial rate of uptake, while for the neutral dipeptide D-Phe-L-Ala and the anionic peptide D-Phe-L-Glu stimulation was observed. However, the effect on D-Phe-L-Glu was more marked than that on D-Phe-L-Ala and the proton activation differed for these two peptides. The calculated Hill coefficients for the two proton-dependent peptides were 1.14±0.16 and 2.15±0.10 for D-Phe-L-Ala and D-Phe-L-Glu, respectively, providing evidence that the stoichiometry of proton: peptide cotransport is different for each peptide (01, 11 and 21 for D-Phe-L-Lys, D-Phe-L-Ala and D-Phe-L-Glu respectively); studies on energetics are compatible with this conclusion. The physiological and molecular implications of this model are discussed, as are the applicability of the conclusions to secondary active transport systems more generally.  相似文献   
3.
4.
定义和区分了化学反应体系中可能存在的两类限制:计量限制和附加限制,从而Gibbs计量规则可用等式I=m-R_3-k表示。着重讨论了附加限制条件以及它对反应体系线性特征的影响,它不影响原子矩阵,但改变计量矩阵和限制矩阵的结构。提出了寻找被修饰的计量矩阵和限制矩阵的方法。  相似文献   
5.
The inclusion complexation of 4-t-butyl-2′-car☐ymethoxy-4′-(3-methyl-2-butenyloxy)chalcone (SU-740), a newly developed antiulcer agent, with β-cyclodextrin (β-CyD) in water and in solid state was investigated for the purpose of improving the low aqueous solubility and oral bioavailability. SU-740 formed 1:1 and 1:2 (guest:host) inclusion complexes in water and in the solid state and the 1:1 stability constant was much higher than the 1:2 stability constant. The dissolution rate of SU-740 was significantly improved by the complexation with β-CyD, suggesting the enhanced bioavailability.  相似文献   
6.
目的 本实验采用HPLC方法建立来自3个产地15批怀牛膝药材的指纹图谱,并对β-蜕皮甾酮,25R-牛膝甾酮,25S-牛膝甾酮进行含量测定,同时通过化学计量学的方法对15批怀牛膝进行质量评价。方法 选用Waters SunFire C18(150 mm×4.6 mm,5 μm)色谱柱,检测波长280 nm,以乙腈-0.1%甲酸水梯度洗脱,流速为1.0 mL·min-1,柱温25℃,进样量10 μL。以β-蜕皮甾酮峰为参照峰,运用软件《中药色谱指纹图谱相似度评价系统(2012版)》对15批怀牛膝药材进行相似度分析,使用Metabo Analyst 5.0网站进行聚类热图分析,使用SIMCA 14.1软件进行PCA和PLS-DA分析。结果 标定了31个共有指纹峰,指认了3个已知成分,并对其进行含量测定,化学计量学分析结果将15批怀牛膝药材样品分为3类,并且筛选了不同产地间潜在的差异性成分。结论 该方法具有良好的精密度、重复性以及稳定性,可为怀牛膝的质量评价与控制提供科学依据及参考。  相似文献   
7.
There is evidence that the production of primary saliva by acinar cells is a consequence of Na+–Cl co-transport but more recently it has been proposed that in fact Na+–K+–2 Cl co-transport is responsible. The latter would be energetically more efficient and the present experiments were designed to measure the stoichiometry of acinar secretion in order to distinguish between these two mechanisms.Submandibular salivary glands from anaesthetised rabbits were isolated vascularly and oxygen consumption measured from the oxygen content of arterial inflow and venous effluent blood and the total flow through the gland. Measurements were made in the steady-state at rest and during different secretion rates induced by parasympathetic nerve stimulation. The rate of sodium transport across the acinar and ductal epithelium was determined from plasma and salivary sodium concentration and salivary flow rate.Multiple regression analysis of this data showed that 22.1 mol Na+ was secreted per mol O2 consumed while 11.9 mol Na+ was reabsorbed per mol O2 consumed. Since acinar secretion is energetically about twice as efficient as ductal absorption, a mechanism for Na+ transport other than that for tight epithelia must be involved. Na+K+–2 Cl co-transport is thus more likely than Na+–Cl and it is suggested that Na+–K+–2 Cl co-transport is the main mechanism involved in salivary acinar secretion.  相似文献   
8.
The coupled transport of Na+ with taurine into snake renal brush-border membrane vesicles (BBMV) was studied using 5-s uptake conditions. Taurine transport into snake renal BBMV involved two parallel processes, one saturable (Na+-dependent) and one (Na+-independent) that behaved like passive diffusion. Below 1 mM taurine concentration, the Na+-dependent system accounted for 60% of total taurine uptake. Over both low (0.001–0.80 mM) and high (0.8–5.0 mM) taurine concentration ranges, the Na+-dependent taurine uptake within each range showed Michaelis-Menten kinetics, suggesting the presence of two independent saturable Na+-dependent transport systems for taurine. The high-affinity, low-capacity system saturated above 100 M with a K m of 71.4±45.7 M and a maximum velocity (V max) of 21.9±3.77 pmol (mg protein)–1 (5 s)–1. The low-affinity, high-capacity system saturated above 1 mM, with a K m of 1.11±0.63 mM and a V max of 252±47 pmol (mg protein)–1 (5 s)–1. The stoichiometric relationship between external Na+ concentration and taurine uptake (at 10 M) by the high-affinity BBMV transport system was examined by the activation method under short-circuited conditions. The 5-s rate of taurine transport was a sigmoid function of increasing extravesicular Na+ concentration. Kinetic analysis of the interaction of Na+ with the high-affinity taurine transport system suggested that 3 Na+ ions (3.2±0.7) may be involved with 1 taurine molecule in the transport event. The data suggest the presence of a highly efficient and high-affinity reabsorptive taurine transport system on the luminal membrane of the kidney of the garter snake, a species that can secrete taurine in vivo.  相似文献   
9.
In order to obtain further information on the interaction between antigens (Ags) and B cell Ag receptors (BCR) for a better understanding of the relationship between signals resulting from Ag binding and B cell activation, effects of Ag valence and size on the apparent association constant, i.e. the avidity as well as the molecular stoichiometry of immune complexes in Ag–antibody (Ab) interactions were studied. Hapten conjugates using proteins of various molecular weights, such as hen egg lysozyme (HEL), ovalbumin (OVA), bovine serum albumin (BSA), and chicken gammaglobulin (CGG), were prepared for this purpose. Different ratios of the hapten, (4-hydroxy-3-nitrophenyl)acetyl (NP), to the protein were used for conjugation, and interactions between anti-NP monoclonal Abs (mAbs) and the NP conjugates were evaluated by surface plasmon resonance. It was founded that the two binding sites of an Ab were able to simultaneously accommodate two NP1-HEL, resulting in a tri-molecular complex, Ag2Ab1. However, NP conjugates of the higher-molecular-weight proteins, OVA and BSA, formed only Ag1Ab1, irrespective of hapten valence. This was thought to be due to steric hindrance caused by the binding of the first Ag. These results suggested that the stoichiometry depended largely on the size of the Ag involved and that mAbs with a low affinity are more efficient at raising the binding strength through divalent interaction since the avidity of two mAbs in interactions with highly haptenated BSA was not significantly different in spite of a 10-fold difference in affinity to the monovalent NP1-HEL.  相似文献   
10.
The TCR/CD3 complex is composed of six subunits which are expressed on the cell surface in a coordinate fashion after assembly in the endoplasmic reticulum (ER). The TCR/CD3 complex is assembled after a series of pairwise interactions involving the formation of dimers of CD3ϵ with either CD3γ or CD3δ. These dimers assemble with TCRα and TCRβ chains, and finally, the CD3ζ homodimer is added to allow export of the full complex from the ER. A model has been proposed suggesting that during assembly the CD3ϵ/CD3γ dimer interacts exclusively with TCRβ and the CD3ϵ/CD3δ dimer with TCRα to form a complex with a single TCRα/β heterodimer. We show in this study, by immunoprecipitation and two-dimensional gel electrophoresis, that in the human T cell line Jurkat as well as in total human thymocytes, this preferential interaction does not occur and instead, the CD3ϵ/CD3γ and CD3ϵ/CD3δ dimers associate with both TCR chains simultaneously and indistinctly. These data are confirmed by the analysis of the TCRα-negative T cell line MOLT-4 in which TCRβ is found separately associated with CD3ϵ/CD3γ and with CD3ϵ/CD3δ dimers. Indirectly, our results support a model of stoichiometry in which two TCRα/β heterodimers are present in a TCR/CD3 complex. Furthermore, immunoprecipitation with anti-CD3γ and anti-CD3δ antibodies from 1 % NP40 and 1 % Brij96 cell lysates showed that these subunits form independent partial complexes which are cross-linked through the CD3ζ homodimer. This suggests that CD3ζ mediates the interaction between both TCRα/β heterodimers contained in the double TCR complex. Further proof for this hypothesis is obtained after analysis of a Jurkat cell transfectant containing a point mutation in the transmembrane domain of TCRβ that impairs the association of CD3ζ. In this mutant cell line, unlike a control line with wild-type TCRβ, the CD3γ- and CD3δ-containing complexes were found completely independent. Altogether, these results support a model of TCR/CD3 assembly and stoichiometry in which two TCR-α/β heterodimers form two hemicomplexes containing either CD3ϵ/γ or CD3ϵ/δ dimers which become associated via the CD3ζ homodimer.  相似文献   
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