On the dielectric relaxation of biological cell suspensions: the effect of the membrane electrical conductivity

Due to the mismatch of the electrical parameters (the permittivity ?' and the electrical conductivity σ) of the membrane of a biological cell with the ones of the cytosol and the extracellular medium, biological cell suspensions are the site, under the influence of an external electric field, of large dielectric relaxations in the radiowave frequency range. However, a point still remains controversial, i.e., whether or not the value of membrane conductivity σ(s) might be extracted from the de-convolution of the dielectric spectra or otherwise if it would be more reasonable to assign to the membrane conductivity a value equal to zero. This point is not to be considered with superficiality since it concerns an a priori choice which ultimately influences the values of the electrical parameters deduced from this technique. As far as this point is concerned, the opinion of the researchers in this field diverges. We believe that, at least within certain limits, the membrane conductivity can be deduced from the shape of the relaxation spectra. We substantiate this thesis with two different examples concerning the first a suspension of human normal erythrocyte cells and the second a suspension of human lymphocyte cells. In both cases, by means of an accurate fitting procedure based on the Levenberg-Marquardt method for complex functions, we can evaluate the membrane conductivity σ(s) with its associated uncertainty. The knowledge of the membrane electrical conductivity will favor the investigation of different ion transport mechanisms across the cell membrane.     

猪小肠抗菌肽对SPF鸡肠道黏膜免疫功能的影响

60只1日龄的SPF 鸡随机分为两组,来评价添加外源性抗菌肽对其肠道黏膜免疫功能的影响.结果表明:1)十二指肠、空肠和回肠内的上皮内淋巴细胞数量从第21日龄起,与对照组相比差异明显(p<0.05).2)自21日龄起,在抗菌肽处理组可以见到明显的肥大细胞,而在对照组少见(p<0.05).3)抗菌肽的处理能显著增加各段肠道内杯状细胞的数量(p<0.05).4)使用抗菌肽处理后,能提高各段肠道的sIgA的表达.添加外源性的抗菌物质,能显著提高SPF鸡的肠道黏膜免疫参数,并且有助于进一步深入了解抗菌肽对肠道黏膜免疫的调控机制.     

PCR扩增人外周血淋巴细胞抗体基因的探讨

采用ＲＴ－ＰＣＲ法,以一组人抗体重链和轻链简并引物,直接从人外周血混合淋巴细胞中扩增出抗体重链Ｆｄ和κ轻链基因．结果提示：用Ｔｒｉｚｏｌ试剂提取细胞总ＲＮＡ,以Ｏｌｉｇ（ｄＴ）引导合成ｃＤＮＡ第一链,再经ＰＣＲ扩增的方法,是值得优先采用的方法．细胞总ＲＮＡ的提取不必追求高纯度,少量外周血即足以满足构建抗体库所需,简并引物可获得良好的扩增效果,为构建噬菌体抗体库奠定了基础     

北沙参免疫活性多糖制备方法研究

为了确定北沙参免疫活性多糖的最佳制备方法,采用4种方法制备北沙参多糖样品,并进行体外小鼠脾淋巴细胞增殖实验,对各样品进行免疫活性比较。以提取率及体外小鼠脾淋巴细胞增殖率为指标,分析95%乙醇回流除去小分子、除蛋白及不同浓度醇沉等步骤的必要性,确定北沙参免疫活性多糖最佳制备方法。通过提取率及体外小鼠脾淋巴细胞增殖率比较,经沸水提取、浓缩和干燥后获得的北沙参免疫活性多糖样品提取率最高,达33.15%;且该样品对小鼠脾淋巴细胞的增殖作用最强,在62.5 μg/mL浓度下增殖率达到33.93%。结果显示北沙参免疫活性多糖的最佳制备方法为沸水提取、浓缩并干燥即可,无需进行95%乙醇回流除去小分子、除蛋白及不同浓度醇沉等步骤。     

胸腺淋巴细胞免疫血清制备的初步研究

试验研究以猪胸腺淋巴细胞为完全抗原,采用一定的免疫程序致敏兔,刺激兔产生抗猪胸腺淋巴细胞的特异性抗体,在30天后分离血清,免疫效价达到1：640,证明兔血清含有该特异抗体,免疫程序合理,抗体效价确实．     

小儿扁桃体淋巴细胞Poly（A）—RNA的制备及其cDNA的合成

采用盐酸胍法从经PHA诱导的小儿扁桃体淋巴细胞中提取总RNA,经Oligo(dT)-纤维素柱层析制备了Poly(A)-RNA。经蔗糖线性密度梯度超速离心步骤,富集了含IL-2mRNA的Poly(A)-RNA。经麦胚无细胞体系翻译实验表明,富集后的Poly(A)-RNA具有良好的模板翻译活性,其~3H-亮氨酸参入活性是对照的5.1倍。又以此富集了的Poly(A)-RNA为模板,按照Watson和Jackson的最新方法合成了小儿扁桃体淋巴细胞的双链cDNA。经碱性凝胶电泳检查,用此方法合成的cDNA长度最长的超过了IL-2的cDNA长度。     

小鼠血清甲状腺激素水平与免疫功能的衰老变化

利用放射免疫测定法和免疫功能指标的检测方法研究了 2 0月龄老年小鼠血清甲状腺激素水平和免疫功能的衰老变化 ,并用外源性甲状腺激素胃内给药的方法探讨了甲状腺机能和免疫功能变化间的因果关系 .结果证明 ,老年期甲状腺机能和免疫功能都有明显的衰退变化 ,且二者间存在有明显的因果关系 .甲状腺机能衰退导致免疫功能紊乱可能是机体衰老的基本过程之一     

富硒米对大鼠细胞免疫功能影响的实验研究

为探讨富硒米的剂量对大鼠免疫功能的影响和富硒米用于保健食疗提供有意义的实验依据,选用纯种ＳＤ大鼠４０只,随机分为高中低三个剂量组,分别给予富硒米和常规饲料喂养３０ｄ,称重,取血做Ｔ淋巴细胞转化和自然杀伤细胞活性试验。结果表明：①富硒米能显著提高大鼠Ｔ淋巴细胞转化率,中剂量组与对照组比较有显著性差异（ｔ检验,Ｐ〈０．０５）。②富硒米大鼠ＮＫ活性对照比较,没有显著性差异 。结论是富硒米能显著提高大鼠Ｔ     

淋巴细胞膜上Na+/Ca2+交换操纵的Eu3+内流的荧光法研究

利用Fura-2荧光浓度指示剂法、通过检测360nm激发荧光强度的变化,研究了Eu³⁺能否利用人外周血淋巴细胞膜上的Na⁺/Ca²⁺交换进入细胞。结果表明:用ouabain预处理细胞无Na⁺介质中测试,当加入Eu³⁺时,360nm荧光强度发生猝灭,且随着胞外加入的Eu³⁺浓度的增大而猝灭增强。表明在实验条件下Eu³⁺可以进入细胞。电压依赖性Ca     

Selective addition of CXCR3+CCR4-CD4+ Th1 cells enhances generation of cytotoxic T cells by dendritic cells in vitro

Increasing importance is being given to the stimulation of Th1 response in cancer immunotherapy because its presence can shift the direction of adaptive immune responses toward protective immunity. Based on chemokine receptor expression, CXCR3⁺CCR4^-CD4⁺ T cells as Th1-type cells were investigated its capacity in monocyte-derived dendritic cell (DC) maturation and polarization, and induction of antigen specific cytotoxic T lymphocytes (CTL) in vitro. The levels of IL-4, IL-5 and IL-10 were decreased to the basal level compared with high production of IFN-γ, TNF-α, and IL-2 in CXCR3⁺CCR4^-CD4⁺ T cells stimulated with anti-CD3 and anti-CD28 antibodies. Co-incubation of activated CD4⁺ or CXCR3⁺CCR4^-CD4⁺ T cells with DC (CD4⁺/DC or CXCR3⁺CD4⁺/DC, respectively) particularly up-regulated IL-12 and CD80 expression compared with DC matured with TNF-α and LPS (mDC). Although there was no significant difference between the effects of the CXCR3⁺CCR4^-CD4⁺ and CD4⁺ T cells on DC phenotype expression, CXCR3⁺CD4⁺/DC in CTL culture were able to expand number of CD8⁺ T cells and increased frequencies of IFN-γ secreting cells and overall cytolytic activity against tumor antigen WT-1. These results demonstrated that the selective addition of CXCR3⁺CCR4^-CD4⁺ T cells to CTL cultures could enhance the induction of CTLs by DC in vitro, and implicated on a novel strategy for adoptive T cell therapy.