Effects of interleukin (IL)-13 on immediate-early response gene expression,phenotype and differentiation of human mast cells. Comparison with IL-4

The recently cloned interleukin 13 (IL-13) shares most investigated biological activities on B lymphocytes and monocytes with IL-4. In this study we investigated the potential role of IL-13 in regulating human mast cell activities. The effects of IL-13 on the expression of an immediate-early response gene (c-fos), proliferation, expression of mast cell-associated cell surface antigen (CD54 and Kit), and in vitro differentiation of human mast cells, were investigated. We compared the effect of IL-13 with that of IL-4. Both IL-13 and IL-4 induced expression of c-fos in cells from the human mast cell line HMC-1. This indicates that mast cells express functional receptors for IL-13. IL-13 and IL-4 decreased the proliferation rate of HMC-1 cells. However, IL-13 was less potent than IL-4. Human mast cells constitutively express the adhesion molecule ICAM-1 (CD54) and the receptor for stem cell factor (Kit) (CD117). The expression of CD54 was increased after treatment with IL-13 or IL-4, whereas the expression of Kit was decreased. Also in this action IL-4 was more potent than IL-13. By culturing mononuclear cells from cord blood in the presence of stem cell factor there is a differentiation of tryptase-positive mast cells in the cultures. This process was inhibited when IL-4 was present. In contrast, IL-13 did not affect the expression of tryptase during differentiation of stem cell factor dependent cord blood-derived mast cells. Taken together, these findings indicate that IL-13 has regulatory effects on human mast cells. The effect overlaps with but is also different from that of IL-4.     

pEGFP/hVEGF121融合蛋白真核表达载体的构建及在骨髓间质干细胞中的表达

目的：构建携带人血管内皮细胞生长因子121及绿色荧光蛋白报告基因的融合蛋白真核表达质粒并检测其在骨髓间质干细胞（MSC）中的表达。 方法： 采用PCR技术，以pCD/hVEGF121质粒为模板扩增VEGF121基因全长，采用PCR产物的粘端克隆法，将VEGF121定向克隆入pEGFP-C1的多克隆位点，构建pEGFP/hVEGF121重组质粒，酶切、PCR及序列分析鉴定，脂质体介导转染体外培养的MSC，荧光显微镜及免疫细胞化学染色检测EGFP/VEGF融合蛋白的表达。 结果： PCR、酶切及测序证实目的基因VEGF121正确连接至pEGFP-C1的多克隆位点，pEGFP/hVEGF121重组质粒转染MSC后，荧光显微镜及免疫细胞化学检测EGFP/VEGF蛋白在MSC中存在表达。 结论： 成功构建了携带人VEGF121及EGFP报告基因的融合蛋白真核表达质粒，并在MSC中获得表达，为进一步研究VEGF基因治疗缺血性心血管疾病及MSC的分化奠定了实验基础。     

脑缺血大鼠大脑皮层和室管膜下区细胞增殖及巢蛋白表达

目的:探讨脑缺血损伤对大鼠脑皮层和室管膜下区细胞增殖及巢蛋白表达的影响。方法:采用大鼠局灶性脑缺血模型,观察大脑皮层和室管膜下区的细胞增殖及巢蛋白阳性细胞表达。结果:脑缺血大鼠室管膜下区细胞增殖明显增加,巢蛋白阳性细胞表达也显著增加;大脑皮层细胞增殖无明显改变,但巢蛋白阳性细胞表达则明显增加。结论:脑缺血损伤可激活室管膜下区与大脑皮层神经干细胞。提示脑缺血大鼠室管膜下区与大脑皮层的神经干细胞可能参与脑缺血损伤的神经修复过程。     

Intrinsic ageing of gut epithelial stem cells

The maintenance of integrity of gut epithelium is essential for the well being of the organism throughout life. Gut epithelium is maintained through a carefully controlled balance between cell loss and renewal, which is sustained by the proliferation of the epithelial stem cells. Although these stem cells show no intrinsic limit to their proliferative capacity, recent evidence indicates that they suffer important functional impairments during the course of ageing. This paper reviews what is currently known about intrinsic ageing of gut epithelial stem cells and its functional consequences.     

Effect of human recombinant erythropoietin on human hemopoietic progenitor cells in vitro

Summary Recombinant human erythropoietin (rhEpo), now available, might become increasingly more important for clinical use, e.g., in the treatment of anemia of chronic renal failure. As a prerequisite of clinical trials, we analyzed the stimulatory and suppressive effects of rhEpo on human hemopoiesis by adding rhEpo to in vitro cultures of nonadherent low-density bone marrow cells obtained from normal persons and from patients undergoing hemodialysis for chronic renal failure. rhEpo was shown to be an effective stimulus for erythroid and multilineage colony formation. The dose-response curve was similar for erythroid progenitors BFU-E from normal controls and patients with chronic renal failure. rhEpo had no effect on megakaryocytic colony formation nor on the megakaryocytic differentiation of multilineage stem cells. Because of a good stimulatory activity on erythroid and multilineage stem cells and lack of toxic effects, rhEpo might be useful in the treatment of certain kinds of anemia.Abbreviations BFU-E burst-forming unit-erythroid - CFU-E colony forming unit-erythroid - CFU-GEMM CFU-granulocyte, erythrocyte, macrophage, megakaryocyte - CFU-GM CFU-granulocyte, macrophage - CFU-Mk CFU-megakaryocyte - IMDM Iscove's modified Dulbecco's medium - Mo-CM Mo-cell line conditioned medium - rhEpo recombinant human erythropoietin     

Morphological features of nerve terminal degeneration as part of the remodeling process in the motor endplate in adult muscles

With the exception of signs of retraction and withdrawal, there have been few morphological data concerning degenerated neural profiles in adult motor endplates. Here, investigation into the ultrastructure of the soleus motor endplates of adult rats (4 months old) turned up particular axonal degeneration in approximately 3% of the subjects. These axons occur as synaptic debris in the synaptic matrix of the motor endplate, adjacent to thin processes of the perisynaptic cells occupying the outer most layer of the motor endplate and were devoid of basal lamina. They often possessed dense-cored vesicles (50-80 nm). Axonal debris released from Schwann cell processes occurred during the period of acute sciatic neurectomy, when nerve terminals progressively disrupted within the motor endplate associated Schwann cells. Finally, immunohistochemical staining for antibodies to label macrophages (ED1 or ED2) has shown that nerve fiber-associated macrophages are located near the motor endplate. The results suggest that during the course of endplate remodeling, a few parts of the terminal branches are disposed of through spontaneous collapse, subsequent release from the Schwann cell investment, and eventual ingestion by macrophages in the perisynaptic space.     

放射损伤对于移植的骨髓间充质干细胞在受体大鼠不同器官中植入的影响

目的： 探讨放射损伤对于骨髓间充质干细胞（MSCs）移植后在受体大鼠不同器官中植入的影响以及可能的机制。方法：采用DNA缺口末端标记法(TUNEL)检测正常对照组和单纯放射组大鼠心脏、肾脏、肝脏、肺脏的细胞凋亡率。应用密度梯度离心法结合贴壁法提取雄性大鼠骨髓MSCs，培养后把MSCs移植到［60COγ］照射和未经照射的雌性大鼠体内，通过PCR和Y染色体荧光原位杂交（Y- FISH）示踪雄性大鼠MSCs在雌性大鼠体内的分布情况。结果：照射后大鼠心脏、肾脏、肝脏、肺脏细胞凋亡率显著高于对照组。PCR结果显示照射后移植MSCs的大鼠心脏、肾脏、肝脏、肺脏和外周血中可以扩增出Sry基因的DNA序列，而且Y- FISH显示照射后移植MSCs的大鼠在心脏、肾脏、肝脏、肺脏中可发现Y染色体阳性的细胞。但是未经照射行MSCs移植的大鼠在上述器官中未发现Y染色体阳性的细胞。结论：全身放射促进组织细胞的凋亡，并导致移植的MSCs在受体大鼠心脏、肾脏、肝脏和肺脏的植入。     

胰岛素样生长因子-1对骨骼肌源性干细胞的促增殖效应

目的 观察胰岛素样生长因子-1(IGF-1)对骨骼肌源性干细胞(MDSCs)生长的影响.方法 采用连续预贴壁法从新生小鼠后肢肌分离培养MDSCs;用含2%胎牛血清的DMEM培养基促进其向骨骼肌细胞分化.免疫细胞化学SP法检测于细胞标志Sca-1和骨骼肌细胞标志肌节(α-sarcomeric)肌动蛋白的表达情况;采用四甲基偶氮唑盐(MTT)比色法检测IGF-1对MDSCs增殖的影响,并分析IGF-1效应与培养时间以及与IGF-1浓度之间的关系.结果 从新生小鼠后肢肌成功分离培养MDSCs,90%以上的MDSCs呈Sca-1阳性;在分化培养中MDSCs能够产生α-sarcomeric 肌动蛋白阳性的肌管;IGF-1对MDSCs促增殖作用随细胞培养时间的延长逐渐明显;随IGF-1浓度的增加而增加,并逐渐趋于饱和.结论 IGF-1对体外培养的MDSCs有促进增殖的作用.     

2型糖尿病并血管病变患者免疫功能研究

目的 :探讨 2型糖尿病并血管病变患者血液中细胞免疫、体液免疫和C反应蛋白 (CRP)的变化。方法 :采用流式细胞术和免疫比浊法测定 42例糖尿病血管病变患者血液细胞免疫参数CD3、CD4、CD8、CD4/CD8、B细胞和NK细胞、血清免疫球蛋白 (IgG、IgA、IgM)、补体 (C3、C4)等体液免疫参数和作为炎症介质的CRP ,并与5 0例 2型糖尿病无血管病变者及 46例正常对照组比较。结果 :2型糖尿病无论是否并发血管病变 ,与正常对照组比较CD3、CD4、CD4/CD8、B、NK显著性减低 (P <0 .0 1) ,IgA、IgM、C3、C4、CRP显著升高(P <0 .0 1) ;并发血管病变组与不并发血管病变组比较C4、CRP显著增高 (P <0 .0 1)。结论 :2型糖尿病患者无论是否并发血管病变 ,机体细胞免疫和体液免疫都异常 ,且炎症介质CRP升高 ,并发血管病变患者机体的炎症介质进一步升高。     

蝎毒多肽对辐射损伤小鼠骨髓造血干细胞及祖细胞的作用

目的探讨不同蝎毒多肽(scorpion venom peptide,SVP)组分对辐射后机体造血干细胞及祖细胞恢复的作用。方法6.0GyX射线一次性全身照射,制作辐射损伤小鼠模型。内源性脾结节法观察照射后第10天脾集落形成单位(CFU-S)的变化。用甲基纤维素半固体培养基培养骨髓混合集落生成单位(CFU-Mix),观察体内外给药方法及照射后不同时间对CFU-Mix生成的影响。结果(1)体内实验:SVPⅣ组分处理后的CFU-S数明显高于照射对照组(P<0.05);SVPⅤ组分CFU-S数量与照射对照组差异无统计学意义。照射后各SVP组CFU-Mix的数量均高于照射对照组,差异有统计学意义(P<0.05)。(2)体外实验:与照射对照组相比,体外分别单独加入SVPⅣ、Ⅴ组分以及细胞因子(IL-6和SCF)均能够促进CFU-Mix的增殖;而SVPⅣ、Ⅴ组分分别与细胞因子联合应用对CFU-Mix生成的促进作用更为明显,其中Ⅳ组分效果更强,与照射对照组相比差异均有统计学意义(P<0.05)。结论SVP具有保护辐射损伤小鼠造血干细胞及祖细胞,加速其增殖能力恢复的作用。