Xenogeneic bone marrow transplantation: II. Porcine-specific growth factors enhance porcine bone marrow engraftment in an in vitro primate microenvironment

Abstract: Establishment of mixed bone marrow chimerism in pig-to-primate transplantation, as a means of inducing specific immune tolerance, will require that both immune and nonimmune barriers be overcome. As a preliminary step in evaluating nonimmune barriers in this system, we have developed an in vitro model of engraftment in which long-term culture of porcine bone marrow-derived hematopoietic cells is supported on preformed primate bone marrow stromal layers. In the absence of cytokine supplementation, primate stromal cells were unable to support long-term porcine hematopoiesis in these cultures. Supplementation with porcine Steel Factor was required for long-term maintenance of hematopoietic progenitor cell content and total hematopoietic activity. Addition of porcine IL-3, in combination with porcine Steel Factor, increased long-term progenitor cell content and hematopoietic activity on primate stroma to levels comparable to that obtained in cultures on porcine stroma. The combination of porcine GM-CSF and Steel Factor increased progenitor cell content and hematopoietic activity early in the cultures, but had little effect in long-term cultures. The Steel Factor and IL-3 combination was species-specific in its action in these cultures, as the corresponding human cytokines were unable to effectively support long-term porcine hematopoiesis. Likewise, the combination of porcine cytokines had only minimal effects on long-term bone marrow culture of primate CD34+ cells I on primate stroma.     

Herpes simplex virus latency in the nervous system – a new model

Permissive herpes simplex virus (HSV) infection in tissue culture results in host cell destruction. Latent HSV infection in vivo occurs in neurons of peripheral sensory ganglia (PSG) and it therefore can not take place in neurons in which the virus has completed a lytic replication cycle similar to that present in vitro. Our hypothesis, based on experimental data and observations in humans, suggests that establishment of latent infection and reactivation of HSV-1 does not involve neuronal cell loss. Latency is established in neurons in which the virus does not replicate and is determined, in part, by the tissue levels of a herpes transactivating protein (Vmw65) that is a component of the viral tegument. We also suggest that reactivation of latent infection does not involve destruction of neurons and is due to replication of virus at the peripheral mucocutaneous tissues to where virus or viral DNA have been transported from the nervous tissue. Alternatively, reactivation is initiated in the PSG using a replication cycle which does not involve irreversible damage to neurons. This model explains the lack of damage to neurons which continue to serve as permanent reservoirs of latent virus for the entire life of the host.     

复合矫治弓丝接头的力学性能及接头组织观察

目的　测试复合矫治弓丝接头的机械性能及分析接头、接头断口变化。方法　测试复合矫治弓丝接头的拉伸强度、剪切强度、扭转强度、弯曲形变残余角,TiNi形状记忆合金丝超弹性,扫描电镜观察及分析接头、断口组织变化。结果　复合矫治弓丝接头抗拉伸强度为32 0～36 0MPa ,剪切强度为2 2 0～2 5 0MPa ,抗扭转强度为30～4 0N·m、断裂时扭转角度不小于1 4 4 0°,弯曲形变残余角为1 2～1 7°;钎焊异质接头中TiNi形状记忆合金丝超弹性几乎没有破坏;接头钎料与TiNi形状记忆合金界面不存在扩散层,与不锈钢丝界面之间有一较宽的扩散层,接头断裂多发生在钎料与TiNi形状记忆合金超弹界面处,部分发生在钎缝中心。结论　复合矫治弓丝机械性能符合正畸临床要求,可应用于临床。     

17β-雌二醇对子宫内膜异位症患者在位子宫内膜间质细胞β-catenin mRNA和蛋白表达的影响

目的研究17β-雌二醇(17β-E2)对子宫内膜异位症(内异症)患者在位子宫内膜间质细胞β-catenin mRNA和蛋白表达的影响,探讨Wnt/β-catenin信号通路在介导雌激素促进内异症发生发展的作用。方法体外分离培养内异症患者在位子宫内膜间质细胞。用不同浓度17β-E2处理子宫内膜间质细胞48 h;此后选用10-10mol/L 17β-E2处理子宫内膜间质细胞12、24和48 h,逆转录聚合酶链反应(RT-PCR)和免疫印迹法(Western blotting)检测17β-E2处理前后子宫内膜间质细胞β-catenin mRNA和蛋白的表达水平。同法分析雌激素受体拮抗剂ICI182,780(10-6mol/L)对17β-E2促进β-catenin mRNA和蛋白表达的影响。免疫组织化学染色观察17β-E2作用后β-catenin在子宫内膜间质细胞中的定位。结果17β-E2能明显促进内异症患者在位子宫内膜间质细胞β-catenin mRNA和蛋白的表达,并呈剂量和时间依赖性,于10-10mol/L作用48 h最明显。雌激素受体拮抗剂ICI182,780能明显抑制17β-E2对子宫内膜间质细胞β-catenin mRNA和蛋白的表达。免疫组织化学染色发现17β-E2能促进β-catenin在子宫内膜间质细胞核内的表达。结论雌激素可能通过激活Wnt/β-catenin信号通路促进内异症在位子宫内膜的异位种植。     

周围神经65KD蛋白单克隆抗体的制备

神经化学诱向生长的研究是神经科学中的一个重要方向。本文以自然系统聚丙酰胺凝胶电泳,分离提取坐骨神经损伤后的远侧端中具有诱神经生长作用的65KD蛋白。以该蛋白作为抗原免疫BALB/C小鼠,通过杂交瘤技术获得一株(VI5E)稳定分泌单克隆抗体的杂交瘤细胞株。免疫印迹法表明,该单克隆抗体特异性地与65KD区带结合。免疫组化法显示,在损伤后的坐骨神经远侧端组织中的雪旺氏细胞呈阳性反应。单克隆抗体的制备为进一步阐明该蛋白的生物学特性奠定了基础。     

Effect of N-tosyl-L-phenylalanylchloromethyl Ketone on Tumor Necrosis Factor-alpha -induced NF-κB Activation and Apoptosis in U937 Cell Line

Summary: To investigate the effect of N-tosyl-L-phenylalanylchloromethyl ketone (TPCK) on tumor necrosis factor-alpha-induced NF-κB activation and apoptosis in U937 cell line, changes and subcellular localization of NF-κB/p65 and IκB-α were observed by fluorescencemicroscopy and expression and degradation of IκB-α by flow cytometry. The apoptosis of U937 cells was measured by flow cytometry and electrophoresis of DNA. Immunolfluorescence assay showed that NF-κB/p65,IκB-α only localized in cytoplasm. After TNF-α stimulation, p65 was localized only in nuclei, and IκB-α was only localized in cytoplasm and decreased. The changes of TNF-α stimulation were specifically inhibited by TPCK. Flow cytometry also revealed the downregulation of IκB-α protein during TNF-α-induced apoptosis and the down-regulation was specifically inhibited by TPCK. Flow cytometry also showed the apoptosis of U937 cells after TNF-α induction. DNA ladder can be detected in cells treated by TNF-α. It is concluded that degradation of IκB-α protein and NF-κB/p65 translocation occur during TNF-α-induced apoptosis of U937 cells, suggesting the activation of NF-κB.TPCK-sensitive protease plays an important role in the degradation of IκB-α protein induced by TNF-α in U937 cells. TPCK sensitive protease also plays an important role in the apoptosis of U937 cells induced by TNF-α.     

The effect of mechanical deformation on magnetic properties and MRI artifacts of type 304 and type 316L stainless steel

The purpose of this study was to evaluate the influence of composition and deformation of biomedical stainless steels on mechanical properties, magnetic properties, and MRI artifacts. Type 304 and Type 316L samples were prepared using standard wire-drawing techniques. Mechanical properties were determined using standard test methods. The amount of ferromagnetic phase present was estimated using a Severn Gage and x-ray diffraction. Magnetic field attraction and artifacts were determined using previously described techniques. The strength of both steels increased significantly with increasing deformation. None of the type 316L wires transformed to the magnetic phase. The amount of magnetic phase in the type 304 wires increased with increasing deformation. There was no magnetic field attraction, and artifacts were minimal for all of type 316L wires and the undeformed type 304 wire. Deflection and artifacts were significant for the deformed type 304 stainless steel. These results provide guidance regarding the use of type 304 and type 316L stainless steels for bioimplants. In this regard, type 316L stainless steel seems to be a more acceptable material with respect to MR compatibility.     

Occupational chronic exposure to metals

Summary Stainless steel welders (n = 103) were examined. To estimate external exposure, personal air sampling was used. Internal exposure was quantified by the determination of nickel levels in erythrocytes, plasma and urine. Men and women (n = 123) were examined for control purposes. In the plasma and erythrocytes of the controls the nickel concentration was below the level of detection (< 1.81 μg/l). The element concentrations in urine were between < 0.1 and 13.3 μg/l. Of the controls 95% showed nickel levels in urine below 2.2 μg/l (reference value). The average concentration of nickel in the air was 93 ± 81 μg/m³. The average concentration of nickel in the plasma samples was 4.9 ± 4.0 μg/l (95th percentile 12.8 μg/L). In erythrocytes nickel could not be detected. The nickel concentrations in the urine of the welders were 18.5 ± 28.5 μg/l on average (95th percentile 52.5 μg/l). Only a weak correlation between the nickel levels of plasma and urine could be detected (C_urine = 2.07 + 8.45 C_plasma; r = 0.294; p < 0.01). Based on our results and on the reported literature a future limit value for the nickel concentration in urine should lay between 30 and 50 μg/l. This value corresponds to an external exposure of 500 μg nickel per cubic metre.     

肉鸡对不同形态锰源的生物利用率研究

目的 :　研究肉仔鸡对不同形态锰源的相对生物学利用率。方法 :　将 62 4只 1日龄肉公鸡随机分为 1 3组 ,分别饲不添加锰的基础饲粮 (对照组 )或以不同锰化合物添加 60、1 2 0、1 80 mg/ kg锰的试验饲粮 2 1 d。结果 :　肉鸡生长性能各指标和腿病发生率以及肉鸡的跖骨灰锰含量、心肌细胞线粒体中 Mn SOD活性均未受到添加锰源以及锰源与锰水平互作的显著影响 ,但受到添加锰水平的显著影响。肉鸡心肌含锰量和 Mn SOD m RNA水平均未受到添加锰源与锰水平互作的显著影响 ,但受到添加锰源及添加锰水平的显著影响。其中中等和强络合强度有机锰源的生物学利用率明显高于无机硫酸锰和弱络合强度有机锰源。结论 :　中等和强络合强度的复合氨基酸锰对肉仔鸡的生物学利用率明显优于无机硫酸锰和弱络合强度的蛋氨酸锰