猪苓多糖、硒及红细胞对LAK细胞杀伤活性的影响

目的　探讨猪苓多糖 (PUPS)、微量元素硒 (Se)及红细胞 (RBC)对LAK细胞杀伤活性的影响。方法　分离正常人外周血单个核细胞 (PBMC)在体外分别与不同浓度的PUPS、亚硒酸钠及红细胞单独或与IL 2协同诱导 ,以MTT法检测LAK细胞活性 ,用ELISA检测培养上清中TNF α和IFN γ含量。结果　不同浓度的PUPS和Se能单独诱导LAK细胞 ,与IL 2协同诱导的LAK细胞活性明显提高 ,细胞分泌的TNF和IFN浓度增高。RBC对LAK细胞的杀伤活性有明显地增强作用 ,并呈浓度依赖关系。结论　PUPS、Se及RBC对LAK细胞杀伤性有明显增强作用 ,细胞杀伤活性的提高可能与内源性TNF和IFN的产生增加有关。本研究为PUPS、Se及RBC与IL 2协同诱导细胞的研究和临床应用提供了理论依据。     

细胞因子诱导破骨细胞分化的2种实验模型的建立

目的探讨体外从单个核细胞诱导分化形成破骨细胞的2种不同方法,建立研究细胞因子对破骨细胞分化作用的2种不同实验模型。方法外周血单个核细胞在10-8mol/L LTB4和25μg/LM-CSF刺激下,经过2周的直接诱导分化;外周血单个核细胞与RAFLs共培养,加入10-8mol/L LTB4和25μg/L M-CSF,经过3周的间接诱导分化,用抗酒石酸酸性磷酸酶(TRAP)染色鉴定破骨细胞样细胞(OCL)。结果通过直接分化和间接分化均可形成破骨细胞样细胞。结论2种不同实验模型能分别用于研究细胞因子对破骨细胞的直接和间接分化作用。     

用干扰素α和内毒素诱导巨噬细胞产生肿瘤坏死因子

本文研究了用IFNα联合LPS诱导小鼠骨髓及腹腔巨噬细胞产生TNF的动态,通过抗TNF血清对TNF活性免疫吸收试验,证明巨噬细胞诱导培养上清引起L929细胞毒作用的因子为TNFα。结果证明:(1)HrIFNα和LPS在诱导骨髓和腹腔巨噬细胞产生TNF上有协同作用。(2)TNFα产生量以培养4hr为最高。(3)小鼠骨髓巨噬细胞为体外诱导TNF产生的较敏感细胞,适于分析各种诱导物诱导TNF产生的效力。     

Tactivin疗法对外伤后骨髓病人IL-1β,TNFα的调节

采用Tactivin疗法对5例下肢外伤后骨髓炎病人治疗前后IL-1β，TNFα的变化进行了观察，对照组为健康人。于Tactivin疗法治疗前后采病人血液，分别检测：①血清IL-1β,TNFα水平；②体外培养外周血单个核细胞自发性和经LPS刺激后分泌的IL-IB，TNFα水平（用免疫酶方法检测）；③外周血单个核细胞的1IL-1β，TNFαmRNA基因表型的检测，用N．Gough方法抽提外周血单个核细胞的mRNA，与用’‘P标记的IL－lpDNA探针、TNFaDNA探针分别进行杂交，用放射自显影方法检测。实验结果表明：①来治疗的骨髓病人血清IL－16水平升高的同…     

人外周血单个核细胞向肝样细胞转化的体外诱导培养方法*

背景：外周血单个核细胞在体外条件下可向肝样细胞转化。 目的：探索体外诱导外周血单个核细胞向肝样细胞分化的培养方法。 方法：采用密度梯度离心法联合贴壁法纯化肝硬化患者外周血单个核细胞,以含巨噬细胞集落刺激因子、白细胞介素3及β-巯基乙醇培养基培养6 d后,诱导组用含肝细胞生长因子,成纤维细胞生长因子4的培养基培养14 d;以未诱导培养的单个核细胞及L02肝脏细胞系为对照。 结果与结论：外周血单个核细胞呈透明圆形、类圆形,用巨噬细胞集落刺激因子、白细胞介素3及 β-巯基乙醇培养基培养6 d后,部分细胞向成纤维样细胞生长,诱导后部分细胞呈多边形,多角形,14 d后细胞形态逐渐接近肝细胞,并表达白蛋白、甲胎蛋白、角蛋白18。说明在一定的诱导条件下,外周血单个核细胞在体外可以向肝样细胞分化。 关键词：肝样细胞;外周血;单个核细胞;诱导;分化;干细胞 doi:10.3969/j.issn.1673-8225.2012.10.018     

雷公藤免疫调节的体外研究

目的证实雷公藤能否选择性地抑制LPS、PHA诱导的TNFα、IFNγ的产生,以及它抑制TNFα的免疫学机理。方法采用健康供血者外周血单个核细胞(PBMC)的体外培养及ELISA,测量雷公藤甲素抑制由LPS、PHA诱导的TNFα、IFNγ的能力;通过对PBMC和单核细胞(monocyte)的预处理试验,以及其后的流式细胞仪分析,研究雷公藤抑制TNFα的免疫学机理。结果雷公藤对TNFα、IFNγ的抑制具有剂量依赖性,对TNFα的IC50为5～10ngml,对IFNγ的IC50为0.1～1.0ngml。PBMC和monocytes试验中,雷公藤的不同预处理后的TNFα浓度,与流式细胞分析产生效应的CD14 TNFα 细胞数一致。结论两种预处理方法提示了不同的临床意义。免疫表现型的分析揭示,雷公藤可能与LPS竞争结合CD14受体。该研究为雷公藤可作为抗炎剂治疗麻风反应提供了基本依据。     

低剂量γ射线辐照外周血单个核细胞对细胞因子表达的影响

目的:探讨低剂量辐射对外周血单个核细胞细胞因子表达的刺激效应.方法:人外周血单个核细胞采用1 Gyγ射线照射,吸收剂量率为17 Gy/min,并在培养过程中不同时间采用ELISA方法检测上清液中IL-2、TNFα及IFN-γ含量.结果:经γ射线照射24 h后培养上清中IL-2、TNFα及IFN-γ含量明显升高,与对照组比较有明显差异(P<0.05),并随培养时间延长,各细胞因子含量会进一步升高.结论:低剂量辐射对外周血单个核细胞中IL-2、TNFα及IFN-γ表达具有刺激效应.     

TNFα对单个大鼠心室肌细胞L型钙电流的影响

目的探讨TNFα对心肌细胞L型钙电流的影响。方法分离大鼠单个心室肌细胞,用全细胞膜片钳方法检测TNFα对单个心室肌细胞L型钙电流的影响。结果 TNFα使单个大鼠心室肌细胞L型钙电流持续增强,促进细胞外钙离子持续向细胞内流动。结论 TNFα与感染性休克心脏收缩性改变相关。     

获得性免疫缺陷综合征病人的抗原特异性多克隆的B细胞应答

获得性免疫缺陷综合征(AIDS)是一种复杂的免疫异常症候群。本文作者检查了58例AIDS和Kaposi氏肉瘤患者的B细胞功能。体外检测结果证明AIDS/KS患者的B细胞在体外失去产生绵羊红细胞(SRBC)抗体的能力。取患者末梢血单个核细胞(PB-MC),用RPMl1640液体外培养,加SRBC进行免疫,用Jerne溶血空斑法测定B细胞     

骨髓基质细胞对多细胞因子介导人脐血单个核细胞体外扩增的影响

背景:体外扩增脐血造血细胞的目的是促进脐血造血细胞的植入能力,细胞因子介导的脐血造血细胞能使细胞数有效扩增,但同时也耗竭了具有分化潜能的造血干细胞。目的:观察骨髓基质细胞对多细胞因子组合介导人脐血单个核细胞体外扩增及扩增后黏附分子和CXCR4表达的影响。方法:将分离的人脐血单个核细胞接种在预先建立的人骨髓基质细胞层上,分组培养:对照组仅含有脐血单个核细胞;多种细胞因子+脐血单个核细胞组;骨髓基质细胞+脐血单个核细胞组;骨髓基质细胞+多种细胞因子+脐血单个核细胞组。培养0,7d检测有核细胞数,CD34+细胞数及CD34+CXCR4+,CD34+CD62L+,CD34+CD49d+的细胞数。结果与结论:单纯骨髓基质细胞和单用细胞因子组均可有效地扩增脐血造血细胞,并提高造血细胞上CD49d,CD62L及CXCR4表达。而单用细胞因子组促进脐血造血细胞扩增能力较单纯骨髓基质细胞强,提高造血细胞CD49d,CD62L及CXCR4表达能力较单纯骨髓基质细胞差。提示骨髓基质细胞虽可支持脐血造血细胞扩增,但难以实现造血细胞的大量扩增,但在骨髓基质细胞层的支持下,多细胞因子可有效地促进脐血造血细胞的扩增,并优于单用细胞因子及骨髓基质细胞,证实骨髓基质细胞可协同多细胞因子有效地促进脐血单个核细胞的扩增,促进造血干细胞的黏附和趋化能力。     

Surface matrix binding alters murine peritoneal mononuclear phagocyte TNF-alpha and IL-6 induction.

Interferon-gamma (IFN-gamma) and bacterial lipopolysaccharide (LPS) are important promotors of mononuclear phagocyte (MO) activation. Signals derived from binding to a surface matrix also participate in promoting the activation process of MO. In this study, we examined the relative contribution of adherence in augmenting murine MO activation for cytokine production. Kinetic studies compared the production and secretion of tumor necrosis factor-alpha (TNF) and interleukin-6 (IL-6) by MO cultured as adherent monolayers to those of MO cultured as suspended cells in teflon vessels. All cells were maximally stimulated in vitro with IFN-gamma and LPS prior to analysis. Immunoprecipitation analysis of protein and RNA slot blots showed that both secreted protein and mRNA representing TNF and IL-6 are delayed two to six hours in nonadherent MO cultures compared to adherent MO cultures. Moreover, data from bioassays confirmed that these cytokines were completely functional in both systems examined. Although IFN-gamma/LPS were able to stimulate production and secretion of TNF and IL-6 in the nonadherent cells, without cell-matrix interaction, the process was significantly delayed. These data support the hypothesis that the physical event of adherence significantly facilitates the production of specific cytokines by activated MO.     

Chemical and morphological studies of liver from eider (Somateria mollissima) in Svalbard with special reference to the distribution of copper

Specimens of male and brooding female eider (Somateria mollissima) were collected in Svalbard. Chemical analyses revealed hepatic copper concentrations ranging from 20 to 1050 micrograms per g wet weight. This is in agreement with previous results. The selenium, zinc and cadmium values were equal to or slightly higher than previously recorded. It is suggested that the wide variation in copper concentration is a result of differences in intake of copper-containing food among the birds. High selenium intake may enhance copper accumulation. Starvation influences the concentration of zinc and also copper. Zinc concentrations were significantly higher in females. This may be secondary to starvation. The percentage of copper recovered among the soluble proteins was inversely related to the copper content. The distribution of the soluble proteins reflects a normal copper metabolism. Microscopic studies showed prominent dark granules, positive with the rubeanic acid test for copper, confined to hepatocytes. By electron microscopy, the granules appeared as large irregular, electron-dense bodies that, by X-ray microanalysis, were found to contain copper. There were no signs of liver injuries such as necrosis and fibrosis. Apparently, the eider has evolved a high capacity for copper storage.     

八肽胆囊收缩素对大鼠滑膜细胞株RSC-364分泌IL-6的影响

探讨八肽胆囊收缩素（CCK-8）对大鼠滑膜细胞株RSC-364分泌IL-6的调控作用。应用ELISA法观察不同浓度梯度的CCK-8对在TNF-α或IL-1β诱导下的RSC-364分泌IL-6水平的影响。结果显示：10^-6mol/L,10^-8mol/L浓度的CCK-8可分别促进TNF-α或IL-1β诱导24h后RSC-364细胞株分泌IL-6，而应用CCK受体拮抗剂丙谷胺则可明显抑制这种刺激效应，提示CCK-8可调节滑膜细胞分泌IL-6，在类风湿性关节炎的发病机制中可能起潜在的调控作用。     

Zinc toxicity in the fowl: ultrastructural pathology and relationship to selenium, lead and copper

Some dietary, enzymic and morphological relationships of selenium, lead and copper to the toxicity of zinc in adult female fowls have been investigated. The anorexia, pancreatic and gizzard lesions which resulted from incorporating 20,000 and 25,000 mg zinc as zinc oxide per kg diet were not prevented by concurrent daily injections of 0.3 mg selenium or the addition of 500 and 2500 mg lead per kg diet. Anorexia and gizzard lesions were also produced by feeding copper sulphate at levels of 2000 and 4000 mg per kg of diet but the pancreas was unaffected. This suggests that zinc may act directly on the pancreas rather than by interfering with the protective function of the selenium. Glutathione peroxidase activity significantly decreased in the blood when zinc-containing diets were fed and selenium injection prevented this. Activity in the liver was also reduced but in the pancreas it was increased by both zinc and zinc plus selenium. Electron microscopy showed that there was an infiltration of heterophils in the gizzard of birds given excess zinc but otherwise there was little qualitative difference in surface cell ultrastructure from the degenerative processes of the normal desquamatory sequence. Pancreatic exocrine cells had reduced numbers of zymogen bodies, distended rough endoplasmic reticulum, many cytoplasmic small, electron-dense bodies and large autophagic vacuoles, sometimes leading to cell death. The acinar lumina were dilated and there was periacinar fibrosis. The significance of these lesions as an indication of zinc-induced interference with pancreatic protein synthesis and membrane integrity is discussed.     

Reduced concentrations of selenium in mild Crohn''s disease.

The concentrations of selenium, zinc, and copper were determined in whole blood, plasma, and leucocytes in 20 patients with relatively mild Crohn's disease, nine of whom were being treated with steroids, and compared with those of a control group. There was a significant decrease in the concentration of selenium in the leucocytes as well as in whole blood and plasma in the patients. Steroids seemed to affect only the concentrations of zinc and copper in plasma. The concentrations of copper and zinc in whole blood, plasma, and leucocytes in patients not taking steroids were not significantly different from those of the control group. The observations suggest that those patients with an apparently satisfactory whole body concentration of copper and zinc may still be at risk of a decrease in the body content of selenium.     

Longitudinal study on trace mineral compositions (selenium, zinc, copper, manganese) in Korean human preterm milk

We measured selenium, zinc, copper and manganese concentrations in the human milk of Korean mothers who gave birth to preterm infants, and compared these measurements with the recommended daily intakes. The samples of human milk were collected postpartum at week-1, -2, -4, -6, -8, and -12, from 67 mothers who gave birth to preterm infants (< 34 weeks, or birth weight < 1.8 kg). All samples were analyzed using atomic absorption spectrophotometry. The concentrations of selenium were 11.8 ± 0.5, 11.4 ± 0.8, 12.7 ± 0.9, 11.4 ± 0.8, 10.8 ± 0.9, and 10.5 ± 1.3 μg/L, zinc were 7.8 ± 0.5, 9.1 ± 0.8, 7.2 ± 0.9, 8.0 ± 0.8, 7.4 ± 0.9, and 6.6 ± 1.2 mg/L, copper were 506 ± 23.6, 489 ± 29.4, 384 ± 33.6, 356 ± 32.9, 303 ± 35.0, and 301 ± 48.0 μg/L and manganese were 133 ± 4.0, 127 ± 6.0, 125 ± 6.0, 123 ± 6.0, 127 ± 6.0, and 108 ± 9.0 μg/L at week-1, -2, -4, -6, -8, and -12, respectively. The concentrations of selenium and zinc meet the daily requirements but that of copper is low and of manganese exceeds daily requirements recommended by the American Academy of Pediatrics, Committee on Nutrition.     

铜,锌对培养心肌细胞缺氧／再给氧损伤的影响

本文观察了不同浓度的铜、锌对培养乳鼠心肌细胞缺氧/再给氧损伤后超氧歧化酶(superoxid dismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-px)活性和脂质过氧化(lipid peroxidation,LPO)代谢产物丙二醛(malondialodehyde,MDA)含量改变,以及乳酸脱氢酶(lactate dehydrogenase,LDH)释放的影响。结果表明:缺氧/再给氧后,心肌细胞SOD活性降低,GSH-px活性和MDA含量增高,LDH释放增加。铜、锌均能不同程度地提高SOD活性,但铜的作用强于锌。锌、铜均可降低LDH的释放,但锌的作用强于铜,锌、铜浓度过高反而增加LDH的释放。此外,铜使GSH-px活性明显升高,并显著降低MDA含量。     

Cross-linking of monocyte plasma membrane Fc alpha, Fc gamma or mannose receptors induces TNF production.

We have studied and compared the effects of IgA and IgG immune complexes and concanavalin A (Con A) on human monocyte tumour necrosis factor (TNF) production. The presence of IgA-containing immune complexes in monocyte monolayers resulted in a dose-dependent increase of TNF production. Similar results were obtained with IgG-containing immune complexes and Con A. The presence of monomeric IgA or IgG did not increase TNF secretion. Both IgA and IgG immune complexes also increased monocyte interleukin-1 beta (IL-1 beta) production. Galactose inhibited the effect of IgA but not IgG immune complexes, while mannose inhibited the effect of Con A. Prednisolone abrogated TNF production, while indomethacin enhanced TNF production in all instances where cross-linking of plasma membrane receptors was achieved. These results indicate that activation of Fc alpha receptors (Fc alpha R), Fc gamma R or mannose receptors of the human monocyte plasma membrane by cross-linking results in increased TNF and IL-1 beta secretion. These findings may be of particular relevance in the pathogenesis of IgA immune complex-mediated disease.     

Suppression of tumor necrosis factor secretion from white blood cells by synthetic antisense phosphorothioate oligodeoxynucleotides

In this ex vivo, rather than in vitro, experiment, a synthetic antisense oligodeoxynucleotide was tested to suppress tumor necrosis factor - alpha(TNF) secretion from lipopolysaccharide-stimulated white blood cells. Antisense oligomer showed significant and specific suppressive effect to the secretion of TNF at concentrations of 1.0 and 10 microM. At the concentration of 1 microM, there were 68.4 and 63.9% suppression of TNF secretion at 2 and 24 h after resuspension of blood cells. At the concentration of 10 microM, the suppressions were slightly higher than those at 1 microM, which were 71.8 and 76.2%, respectively. A 50%-matched scrambler showed suppressive effect only at 10 microM concentration, and the suppression only occurred at 2 and 24 h after incubation. Sense oligomer showed no suppressive effects at any of the concentrations. The specificity of this oligomer was documented by dose-effect phenomenon, sequence-dependent suppression and absence of effect on the synthesis of another cytokine (interleukin-6). A series of parallel studies was performed and showed that all three oligomers at any concentration tested had no effect on the interleukin-6 secretion after LPS stimulation.In conclusion, properly designed antisense oligodeoxynucleotide can significantly and specifically suppress the secretion of TNF by blood cells in an ex vivo system and it may be a good "information" drug to treat diseases that are caused by over production of TNF.