自体骨髓干细胞移植治疗神经系统损伤和变性疾病42例报告

目的:观察42例神经系统损伤和变性疾病患者经自体骨髓干细胞移植后,其神经系统症状与体征的改善。方法:①选取2005-06/2006-09解放军第四六三医院神经外科收治的神经系统损伤和变性疾病患者42例,对本试验均签署知情同意书。其中各种原因所致脊髓损伤30例,脑血管意外引起的偏瘫4例,老年痴呆2例,小脑疾患引起的共济失调2例,运动神经元变性疾病4例。患者入院后常规检查身体各项指标以评估是否适合行干细胞移植术。②术前患者均于皮下注射粒细胞集落刺激因子进行骨髓干细胞动员,5～12μg/(kg.d),连续4～5d。动员结束后患者在局麻状态下,于髂后上棘采集自体骨髓血约200mL,去除红细胞,密度梯度离心,分离出单个核细胞,再将收获的单个核细胞制成1.5mL细胞悬液(约1×108个细胞)备用。③通过手术或立体定向的方法将干细胞直接移植在神经损伤或变性部位,然后再根据病情经腰穿或静脉途径进行细胞移植,每次给予干细胞量按(2～3)×102个/kg计算,1次/周,共4周。④自体骨髓干细胞移植治疗过程中,为促进干细胞的生长和分化,根据患者病情及个人身体状况给予物理疗法、作业疗法等相应的康复功能锻炼,同时配以针灸、高压氧及药物营养神经和改善微循环治疗。结果:42例神经系统损伤和变性疾病患者均进入结果分析。自体骨髓干细胞移植治疗后1～3个月,17例脊髓损伤患者肢体症状明显改善,尤其是脑损伤后遗症引起的脊髓损伤其肢体症状改善更为显著;2例脑出血和1例脑血栓引起偏瘫的患者其肢体症状改善效果均较好;遗传性共济失调的2例患者疗效均不明显;1例老年痴呆患者其老年痴呆症状得到明显改善,并能在搀扶下行走,与旁人进行正常的交流;运动神经元变性病中1例患者肌萎缩性侧索硬化症改善效果较好,肌张力正常,颤抖减轻及至消失,多发性硬化中2例患者症状减轻。结论:干细胞移植配合功能锻炼等辅助手段对神经系统损伤和变性疾病的治疗效果肯定,有一定的临床应用价值。     

卟啉化合物的研究:取代四苯基卟啉与5-氟脲嘧啶共价相连化合物的合成及其抗癌活性

合成了3个取代四苯基卟啉与5-氟脲嘧啶以共价相连的化合物。通过用它们加光照对小鼠移植瘤生长的抑制作用、对小鼠癌细胞的杀伤效果及对小鼠耳指数的测定,初步结果显示,化合物1,2,3对癌细胞都有光敏毒杀效应,但1远较2和3显著。     

Toxicity to fathead minnows of endrin in food and water

Fathead minnows (Pimephales promelas) were exposed during a partial chronic toxicity study to endrin concentrations in the water or food, or both, for 300 days encompassing reproduction. Tissue residues were analyzed at preset intervals for first-generation fish, and were also determined for embryos, larvae at hatch, and 30-day progeny. Gas-chromatographic and liquid-scintillation techniques were used to monitor the contribution of endrin from each source. The food was clams that had accumulated 14C-endrin when exposed to an endrin water concentration similar to that to which the fish were exposed. Higher endrin tissue residues were accumulated from the water than from food. Maximum concentration factors were 0.8 from the food and 13,000 from the water. Residues contributed by endrin in the food were additive to those from the water at all life stages. Endrin in the food (0.63 ppm) significantly reduced survival of the fathead minnows, and fish exposed to both endrin sources had lower survival than those exposed to either source alone. Endrin residues in embryos and larvae were highest and larval survival lowest for progeny of adults exposed to endrin in both food and water. Survival of 30-day progeny was significantly reduced at all test exposures (0.63 ppm in the food, water exposures of 0.14 and 0.25 ppb, and all combinations of food and water exposure).     

CD154 on the surface of CD4+CD25+ regulatory T cells contributes to skin transplant tolerance

BACKGROUND: It is known that the infusion of whole blood from donors (donor-specific transfusion) into recipients combined with anti-CD154 therapy can prolong allograft survival. It has generally been agreed that the effectiveness of anti-CD154 therapy is caused by the inactivation of alloreactive CD4+ and CD8+ effector T cells. The recent literature has implicated CD4+CD25+ regulatory T cells in the suppression of autoimmunity and graft rejection, and we therefore examined whether CD154 blockade is effective because of its blockade of inflammatory T-cell activation or because of a direct impact on the regulatory T cells. METHODS: RAG(-/-) mice were adoptively transfused with CD4+ T cells or a subset of the population (CD4+CD25+ or CD4+CD25- T cells) alone or in combination with donor-specific transfusion and anti-CD154 and given an allo-skin transplant. The longevity of the transplant was determined over time. CD154(-/-)CD4+ T cells were used to assess the importance of CD154 in graft rejection and acceptance. RESULTS: CD154 blockade (or loss of CD154) on CD4+CD25+ regulatory T cells enhanced their immunosuppressive activities and was a contributing factor to anti-CD154-induced immune suppression in vivo. In a model of allograft tolerance, suppression was elicited by antigen and anti-CD154 or antigen alone if the CD4+CD25+ regulatory T cells were deficient in CD154 expression. CONCLUSIONS: Neutralizing the function of CD154 on regulatory T cells upon antigen exposure induces heightened levels of suppressive activities and is likely a contributing factor to the long-lived therapeutic effects of anti-CD154 treatment.     

葛根素对大鼠成骨细胞增殖和分化的影响

目的:课题以往的研究已证实,葛根素在体外能够抑制破骨细胞的骨吸收活性,促进骨生长,实验将进一步验证葛根素对大鼠成骨细胞增殖和分化的影响,并评价其对骨形成的作用。方法:实验于2003-03/2004-03在武装警察部队医学院药物化学教研室药物活性筛选室进行。①实验材料:葛根素(Puerarin)购自中国药品生物制品检定所,雌酚酮购自浙江仙居制药厂。大鼠由解放军军事医学科学院实验动物中心提供。②实验方法:实验分5组:雌酚酮阳性对照组、空白对照组(不加药,只加等量细胞悬液和培养基)、0.01,0.1,1μmol/L葛根素组。由新生24hSD大鼠颅盖骨分离培养成骨细胞,调整成骨活细胞密度为0.9×107L-1,接种于24孔培养板,每组12个复孔,分别于第1,3,5天每组取3孔计数,绘制细胞生长曲线。③实验评估:分别于第1,3,5天采用四唑盐法测定吸光度,计算平均增殖率,采用硝基苯基质动力学法测定各组细胞内外碱性磷酸酶活性。结果:①与空白对照组相比,0.1,1μmol/L葛根素干预组与雌酚酮阳性对照组第3,5天细胞增殖速度均加快,差异有显著性意义(P<0.01)。②经1μmol/L葛根素处理的第1,3,5天,细胞内、外碱性磷酸酶活性均明显高于空白对照组(P<0.01)。经0.1μmol/L葛根素处理第3,5天,细胞内、外碱性磷酸酶活性明显高于空白对照组(P<0.05,P<0.01)。经0.01μmol/L葛根素处理后第5天,细胞外碱性磷酸酶活性明显高于空白对照组(P<0.01)。结论:葛根素对骨形成的作用:①通过影响碱磷酸酶活性,而促进成骨细胞分化。②通过雌激素受体介导促进成骨细胞的骨形成效应。     

嗅鞘细胞的特点及其研究现状

目的:嗅鞘细胞具有强大的修复脊髓损伤的潜能,文章就嗅鞘细胞的特性、培养及纯化方法、移植时机选择、应用前景等内容予以综述。资料来源:应用计算机检索Medline 1994-01/2007-03期间的相关文章,检索词为“olfactory ensheathing cells,spinal cordinjury”,并限定文章语言种类为English。同时计算机检索中国期刊全文数据库1994-01/2007-03期间的相关文章,检索词为“嗅鞘细胞,脊髓损伤”,并限定文章语言种类为中文。资料选择:对资料进行初审,并查看每篇文献后的引文。纳入标准:文章所述内容应与嗅鞘细胞的特性、培养、移植、应用等研究相关。排除标准:重复研究。资料提炼:共收集到47篇相关文献,35篇文献符合纳入标准,排除的12篇文献为内容陈旧或重复。符合纳入标准的35篇文献中,分别涉及嗅鞘细胞的特性、培养及纯化方法、移植时机选择、应用前景等内容。资料综合:脊髓损伤后脊髓神经元死亡,神经营养因子缺乏,且存在抑制修复分子以及胶质瘢痕、空洞阻碍轴突生长。脊髓损伤修复的策略就是针对这些因素加以干预,但完全恢复脊髓功能非常困难且并非必要,治疗目的是恢复对生活质量有重要作用的那一部分功能,优先考虑的重点是髓鞘形成和轴突再生。嗅鞘细胞能分泌多种神经营养因子,并抑制星形胶质细胞反应,具有明显的神经保护作用,加之其强大的增殖能力,在细胞移植和基因治疗等研究领域存在许多优势。嗅鞘细胞取材方便、易于扩增,可作为脊髓损伤细胞治疗的一个备选来源。结论:具有自我更新能力及神经保护作用的嗅鞘细胞,随着转基因治疗和联合细胞移植的深入研究,将为脊髓损伤的修复带来广阔前景。     

左旋精氨酸与褪黑激素对大鼠肾脏缺血再灌注损伤的保护作用

目的:诱导型一氧化氮合酶来源的一氧化氮与内皮型一氧化氮合酶来源的一氧化氮的平衡对维持血流动力学稳定及内环境动态平衡有重要作用.观察左旋精氨酸与褪黑激素对肾脏缺血再灌注损伤中诱导型一氧化氮合酶与内皮型一氧化氮合酶的影响.方法:实验于2006-07/2007-03在中山大学医学院动物实验中心完成.①实验分组:健康雄性清洁级SD大鼠60只,体质量170～210 g,随机数字表法分为缺血再灌注组、左旋精氨酸治疗组、褪黑激素治疗组、联合治疗组,每组15只.②实验方法:建立大鼠肾脏缺血再灌注损伤模型,术前1 h各组大鼠腹腔内注射生理盐水及相应药物,每隔24 h重复注射1次,连续5d.③实验评估:观察各组术前及术后1,3,5 d血肌酐、诱导型一氧化氮合酶和内皮型一氧化氮合酶的变化规律及肾脏病理组织学的改变,并采用Paller法对术后第3天肾小管进行评分.结果:纳入大鼠60只,均进入结果分析.①血肌酐的表达:术后第1天联合治疗组水平显著低于缺血再灌注组(P＜0.05);术后第3,5天左旋精氨酸治疗组、褪黑激素治疗组与联合治疗组显著低于缺血再灌注组(P＜0.05).②内皮型一氧化氮合酶的表达:术后第3天缺血再灌注组比术前降低(P＜0.05);术后第3天和第5天左旋精氨酸治疗组、联合治疗组高于缺血再灌注组(P＜0.05).③诱导型一氧化氮合酶的表达:术后第1天缺血再灌注组与术前相比升高,第3天达到高峰,第5天仍高于术前(P＜0.05);褪黑激素治疗组、联合治疗组术后第3天与第5天明显低于缺血再灌注组(P＜0.05).④血肌酐水平与诱导型一氧化氮合酶的表达呈正相关,r=0.57,P＜0.01,与内皮型一氧化氮合酶/诱导型一氧化氮合酶呈负相关,r=-0.61,P＜0.01.⑤肾小管损伤Palller法评分:依次为联合治疗组＜褪黑激素治疗组＜左旋精氨酸治疗组＜缺血再灌注组.结论:通过调高内皮型一氧化氮合酶的表达、抑制诱导型一氧化氮合酶的表达可以减轻肾脏缺血再灌注损伤,联合应用左型精氨酸与褪黑激素即通过以上途径更有效的起到对肾脏缺血再灌注损伤的保护作用.     

服刑人员132例人格障碍筛查问卷调查

目的:了解人格障碍筛查问卷的初步诊断情况。方法:于2005-09/10选择广东省三水劳教所的服刑人员154人为调查对象,将人格障碍筛查问卷的软件中的106个检查条目制成“是”或“否”问卷进行调查。结果:发放问卷154份,回收问卷154份,有效问卷132份。132名服刑人员中未提示任何人格障碍和品行障碍的有10名(7.58%)。73名提示18岁前有品行障碍,提示有人格障碍的116名,99名存在2项以上人格障碍。结论:问卷结果提示的人格障碍和品行障碍有可能存在诊断过度的问题。对调查阳性者进行人格障碍临床半定式检测工具测试是下一步要进行的工作。