藏药结血蒿水提物的免疫抑制作用研究

目的研究结血蒿水提取物（AV-ext）对小鼠免疫反应的抑制作用。方法运用淋巴细胞增殖反应、混合淋巴细胞反应观察AV-ext体外应用和口服给药对小鼠脾细胞增殖的影响;运用小鼠脾细胞增殖试验,观察AV-ext体外应用对小鼠活化脾细胞分泌IL-2的影响;用明胶酶谱法和粘附分析法,观察AV-ext对小鼠活化T淋巴细胞移动和粘附能力的影响。结果AV-ext在1μg／mL-100μg／mL剂量范围内对脾细胞无明显的细胞毒性,但对刀豆蛋白A（ConA）诱导的小鼠脾细胞增殖反应、混合淋巴细胞反应具有明显的抑制作用,对小鼠活化脾细胞分泌IL-2、小鼠活化T淋巴细胞分泌基质金属蛋白酶9（MMP-9）及粘附能力有显著的抑制作用;连续7d每日口服给予150mg／kg和300mg／kg的AV-ext,能明显抑制小鼠脾细胞增殖及活化脾细胞分泌MMP-9。结论AV-ext能显著抑制小鼠的细胞免疫反应。     

HIV-1C亚型nef基因构建的DNA质粒诱导小鼠免疫反应的研究

目的构建表达中国流行株HIV-1C亚型调控nef基因的重组质粒pVAX-nef，并免疫BALB／c小鼠，观察其免疫效果，为探索新型HIV DNA疫苗提供数据。方法利用分子生物学技术，将nef基因克隆到pVAX，并在体外进行表达与鉴定。以纯化的Nef蛋白作为包被抗原，用ELISA检测其体液免疫反应，用ELISPOT检测其细胞免疫反应。结果重组质粒pVAX-nef成功构建。接种小鼠后2周，ELISPOT结果显示产生了针对HIV特异的CD4和CD8细胞抗原表位的免疫应答，且与免疫剂量存在一定的正相关性。ELISA实验诱导产生了抗HIV-1特异性抗体，其中40μg免疫组诱导的抗体水平最高。结论重组质粒pVAX-nef免疫小鼠后可有效地诱导机体产生细胞免疫和体液免疫反应。     

热应激蛋白70的研究进展

热应激蛋白70(HSP70)是由细胞核内高度保守的热应激基因编码的产物之一,作为对外界应激的一种保护性反应,它有助于机体维持正常的新陈代谢和细胞结构的完整性。当前,HSP70的研究已成为当今世界生命科学的热点和富有希望的研究领域。本文就其结构与功能、基因表达的调控以及应用前景作一综述。     

冷冻异体神经移植应用转化生长因子β1质粒的研究

目的探讨对经反复冻融处理的冷藏异体神经移植,局部使用转化生长因子β1(transforming growth factor β1,TGF-β1)质粒的效果. 方法健康成年不同窝Wistar大鼠40只随机分为两组,每组20只,将坐骨神经在梨状肌孔下5 mm处切除2.0 cm,神经缺损用预制反复冻融于-196℃冷冻的异体神经2.0 cm移植修复.实验组在局部肌肉及神经两断端注射TGF-β1质粒,对照组注射等量生理盐水.分别于6周和12周对每组10只大鼠取材,切片、染色并进行轴索计数和统计学分析. 结果 6周时实验组轴索间基本无水肿,再生神经数量接近正常;对照组神经移植段轴索间有轻度水肿,再生神经较少.12周时实验组整个神经移植段基本被再生轴突充满,有髓纤维排列整齐,轴突和髓鞘发育良好,再生轴突数为98.6±4.8/μm2;对照组神经纤维排列整齐,轴突和髓鞘发育较实验组差,轴突数为75.8± 5.1/μm2,差异有统计学意义(P<0.01). 结论反复冻融冷藏保存的异体神经其抗原性降低,具有修复神经缺损的可能性.局部使用TGF-β1质粒可发挥免疫抑制作用,降低宿主的免疫排斥反应.     

刺五加注射液对小鼠移植性实体型宫颈癌(U_(14))影响的初步观察

用(?)五加注射液注入移植性宫颈癌荷瘤小鼠体内后,观察了不同时期小鼠体内肿瘤重量及癌周淋巴一浆细胞的密度变化。结果表明:刺五加注射液对小鼠肿瘤的生长无明显影响,但有明显的增强癌周淋巴细胞一浆细胞反应的作用。     

全身性非特异性免疫反应对大鼠睾丸功能的影响──Ⅱ.激素水平的变化

本实验以ＳＤ雄性大鼠为实验对象，采用碳粒血管内注射的方法，给动物造成一种全身性的非特异性免疫反应状态，以观察在这种状态下睾丸和垂体的激素分泌情况。动物经每天１次，共１０天的碳粒鼠尾静脉注射后，血浆睾酮的水平（０．８９６±０．３５８ｎｇ／ｄｌ，ｎ＝５）明显低于对照组（２．６５６±０．９９３ｎｇ／ｄｌ，ｎ＝７，Ｐ＜０．００５）；血浆ＬＨ（３．６７６±１．３５０ｍＩＵ／ｍｌ，ｎ＝９，４．６２７±２．５３９ｍＩＵ／ｍｌ，ｎ＝１０）两组比较无显著差异（Ｐ＞０．０５）；ＦＳＨ实验组（３．３６２±０．９２６ｍＩＵ／ｍｌ，ｎ＝９）与对照组相比明显降低（４．８９４±１．２３６ｍＩＵ／ｍｌ，ｎ＝１０，Ｐ＜０．０１）。实验组动物睾丸间质内ＡＣＰ阳性反应细胞增多，而β－羟基甾体脱氢酶反应消失。作者认为全身性的免疫反应既可以通过垂体也可以直接的作用于睾丸内的相关细胞，从而影响睾丸的功能。     

天狼猩红-偏振光法检测子宫内膜异位症中Ⅰ、Ⅲ型胶原

探讨子宫内膜异位症中Ⅰ、Ⅲ型胶原的含量和作用。方法：对子宫内膜异位症（内异症）２０例中的异位内膜组织红色病变期１２例（Ａ组）、黑色病变期８例（Ｂ组）和１１例非子宫内膜异位症患者的在位子宫内膜组织（Ｃ组）用天狼猩红染色,偏振光观察,ｐｈｏｔｏｓｈｏｐ分析,半定量研究Ⅰ、Ⅲ型胶原的含量。结果： Ｉ、Ⅲ型胶原在Ｃ组中含量最低（Ｐ＜０．０５）,Ⅲ型胶原在Ａ组中含量最高（Ｐ＜０．０５）,Ｉ型胶原在Ｂ组中含量最高（Ｐ＜０．０５）。Ｉ、Ⅲ型胶原在增生期和分泌期差异无显著性（Ｐ＞０．０５）。结论：Ⅰ、Ⅲ型胶原的异常沉积可引起内异症患者腹腔内发生异常的免疫反应。     

Osteogenic differentiation of bone marrow MSCs by β-tricalcium phosphate stimulating macrophages via BMP2 signalling pathway

Immune reactions play important roles in determining the in vivo fate of bone substitute materials, either in new bone formation or inflammatory fibrous tissue encapsulation. The paradigm for the development of bone substitute materials has been shifted from inert to immunomodulatory materials, emphasizing the importance of immune cells in the material evaluation. Macrophages, the major effector cells in the immune reaction to implants, are indispensable for osteogenesis and their heterogeneity and plasticity render macrophages a primer target for immune system modulation. However, there are very few reports about the effects of macrophages on biomaterial-regulated osteogenesis. In this study, we used β-tricalcium phosphate (β-TCP) as a model biomaterial to investigate the role of macrophages on the material stimulated osteogenesis. The macrophage phenotype switched to M2 extreme in response to β-TCP extracts, which was related to the activation of calcium-sensing receptor (CaSR) pathway. Bone morphogenetic protein 2 (BMP2) was also significantly upregulated by the β-TCP stimulation, indicating that macrophage may participate in the β-TCP stimulated osteogenesis. Interestingly, when macrophage-conditioned β-TCP extracts were applied to bone marrow mesenchymal stem cells (BMSCs), the osteogenic differentiation of BMSCs was significantly enhanced, indicating the important role of macrophages in biomaterial-induced osteogenesis. These findings provided valuable insights into the mechanism of material-stimulated osteogenesis, and a strategy to optimize the evaluation system for the in vitro osteogenesis capacity of bone substitute materials.     

Expression of ciliary neurotrophic factor is maintained in spinal motor neurons of amyotrophic lateral sclerosis

Ciliary neurotrophic factor (CNTF) was originally identified as a potent survival factor for a variety of neuronal cell types in vitro and in vivo and in particular in spinal motor neurons of embryonic chick and rat. Using a monoclonal antibody against CNTF (clone 4–68) we analysed the expression of CNTF in paraffin sections of seven human brains and spinal cords immunocytochemically using the ABC method and compared these results with sections of the spinal cords of patients suffering from amyotrophic lateral sclerosis (ALS). In normal human tissue of the central nervous system CNTF immunoreactivity was found in most of the motor neurons of the motor cortex and ventral horn, neurons of the nucleus oculomotorius, intermediolateralis, thoracicus, ependymal cells as well as in smooth muscle cells and endothelial cells of small arteries. A reduced number of astrocytes showed a positive immunocytochemical reaction. In peripheral nerves and nerve roots of the spinal cord we also found a positive staining of Schwann cells and some axons. These immunoreactions could be confirmed by Western blot analyses. Next we analysed postmortem paraffin sections of the spinal cord of seven patients suffering from ALS (age range 30–76 years, median age 46 years, female/male = 4:3). We found CNTF immunoreactivity in most of the motor neurons of the ventral horn in 5 cases. In two cases the number of positively stained motor neurons was less. From these results we conclude that CNTF is expressed in a high number of upper and lower motor neurons in the human CNS and that its expression is maintained in ALS patients.