纤维环穿刺法与纤维环切开法建立兔椎间盘退变模型

背景：椎间盘退行性变模型的建立,是研究椎间盘退行性变病理过程和尝试基因治疗等的基础,建立的动物模型要求与人类椎间盘退行性变具有相似性和可比拟性,但目前尚缺乏公认的最佳实验动物模型。 目的：比较纤维环穿刺法和纤维环切开法建立兔椎间盘退变动物模型的差异。 方法：将32只新西兰大白兔以数字表法随机分为纤维环穿刺组和纤维环切开组。经腹膜外入路暴露L3/4、L4/5、L5/6椎间隙,纤维环穿刺组采取针刺纤维环,纤维环切开组采取尖刀切开纤维环,控制穿刺或切开的深度及方向。术后2,4,12,20周通过MRI和组织病理学检查观察腰椎间盘髓核变性及组织病理情况。 结果与结论：术后4周,兔椎间盘髓核面积缩小,纤维环面积增大,髓核内T2加权像信号降低、变暗,椎间隙高度也开始下降,纤维环穿刺组T2信号强度评分较纤维环切开组低(P < 0.05);随着时间的进展,兔椎间盘T2信号强度评分逐步增高,椎间隙逐渐变窄,术后20周椎间盘T2信号强度评分达最高,两组比较差异无显著性意义。随着时间的进展,两组髓核内细胞含量逐渐减少,纤维软骨形成。提示纤维环穿刺法和纤维环切开法均可成功建立椎间盘退变模型,但纤维环切开法椎间盘的退变程度较纤维环穿刺法剧烈,建立的模型过程可能不是人体椎间盘自然退变的模拟过程,纤维环穿刺法比较真实地模拟了人类椎间盘损伤后的退变过程。     

多效生长因子及血小板源性生长因子在增龄鼠椎间盘中的表达*

背景：在组织缺氧状态下,多效生长因子能够直接参与新生血管的生成,在人类腰椎间盘退变方面起重要的作用。 目的：观察增龄鼠椎间盘中多效生长因子和血小板源性生长因子的表达,及其与椎间盘退变的关系。 方法：取1,3,6,12,18月龄Wistar大鼠各10只。采用免疫组织化学染色方法检测椎间盘组织中多效生长因子及血小板源性生长因子的表达情况。采用Spearman相关分析方法对多效生长因子和血小板源性生长因子的相关性进行分析。 结果与结论：随着大鼠月龄的增加,椎间盘细胞中多效生长因子及血小板源性生长因子的表达有逐渐减低的趋势,但至18月龄又有所增加。同月龄组椎间盘细胞中,多效生长因子及血小板源性生长因子在终板的表达高于髓核和纤维环(P < 0.05)。多效生长因子及血小板源性生长因子在椎间盘组织中的表达有一定的相关性(P < 0.01)。提示大鼠椎间盘结构随月龄增加发生退行性变,多效生长因子及血小板源性生长因子参与了大鼠椎间盘的退变。     

经皮纤维环穿刺法建立兔椎间盘退行性变动物模型

背景：合适的椎间盘退变动物模型是椎间盘组织工程研究的必要条件,但目前尚缺乏公认的模型制备方法。 目的：采用C形臂辅助下经皮纤维环穿刺法制备兔椎间盘退变动物模型,并评估其可行性。 方法：选定新西兰大白兔L2/3和L3/4椎间盘作为穿刺干预椎间盘,L1/2和L5/6椎间盘作为空白对照组,采用C形臂辅助下经皮穿刺法干预椎间盘。于术后2,4,8,12周各选择2只兔麻醉后拍摄兔腰椎核磁共振影像,处死动物采集椎间盘,进行椎间盘髓核蛋白多糖测定。核磁共振检查观察椎间盘退行性改变,二甲基亚甲蓝染色分光光度法测定髓核中蛋白多糖含量变化。 结果与结论：术后4周穿刺干预椎间盘髓核区域核磁共振信号强度及髓核内蛋白多糖含量同空白对照组相比均下降(P < 0.05),其后两者呈现逐渐下降趋势。结果证实,C形臂X射线机辅助下经皮纤维环穿刺法可用于椎间盘退变动物模型的制备。     

腰椎运动节段终板凹陷角变化的有限元分析*☆

背景：终板的组织形态改变可通过影响对椎间盘的营养传递最终导致椎间盘的退变。 目的：观察终板凹陷程度变化对腰椎运动节段生物力学的影响。 设计、时间及地点：有限元模型生物力学分析。于2005-01/2007-01在浙江大学医学院附属第二医院骨科生物力学实验室完成。 材料：德国西门子SOMATOM SENSATION 16螺旋CT机。ANSYS有限元分析软件(Inc. Pennsylvania, USA)。 方法：在以往建立的腰椎L4~5运动节段三维非线性有限元模型基础上，采用CAD方法精确构建大、中、小3种不同终板凹陷角的有限元模型，有限元模型的椎间盘前凸角、小关节间隙等其余形态学参数及网格划分均保持一致。垂直压缩、屈曲、伸直、前后剪力5种载荷条件下，分别对3种有限元模型生物力学参数进行测试。 主要观察指标：终板-椎间盘界面应变、椎间盘刚度、髓核内压、椎间盘膨出、纤维环纤维张应力、纤维环基质应力、腰椎后部结构应力以及关节突接触力。 结果：负载条件下，终板凹陷角增加、终板凹陷程度减小可导致终板-椎间盘界面应变减小，椎间盘刚度及髓核内压增加，椎间盘膨出、纤维环纤维张应力、纤维环基质应力、腰椎后部结构应力以及关节突接触力减小。 结论：终板凹陷程度的减小增强了椎间盘对椎体的保护作用；同时可通过影响终板的形变减少对椎间盘的营养传递。     

人颈椎间盘退变与细胞凋亡及基质金属蛋白酶11的表达**☆

背景：前期研究发现基质金属蛋白酶11基因在人退变颈、腰椎间盘组织中明显上调。 目的：观察人退变颈椎间盘髓核组织中基质金属蛋白酶11的表达与细胞凋亡的关系。 方法：纳入30个经MRI确认的退变颈椎间盘髓核组织和20个因颈椎创伤治疗获得的正常颈椎间盘髓核组织。 结果与结论：苏木精-伊红染色显示退变的颈椎间盘髓核组织中髓核细胞较正常髓核组织明显减少(P < 0.01),而凋亡细胞较正常髓核组织明显增多(P < 0.01)。免疫组化染色显示退变的颈椎间盘髓核组织中基质金属蛋白酶11的表达明显高于正常髓核组织(P < 0.01),且基质金属蛋白酶11表达与TUNEL染色检测到的细胞凋亡正相关(r=0.44,P < 0.05)。说明高表达的基质金属蛋白酶11不仅可直接破坏细胞外基质尚可诱导髓核细胞凋亡,在椎间盘退变的过程中发挥重要作用。     

核因子κB、基质金属蛋白酶3在退变腰椎间盘组织中的表达*

背景：对于椎间盘退变的原因一直是国内外学者研究的重点。近年来随着研究的深入,在椎间盘退变过程中多种细胞因子、黏附分子、趋化因子、炎症递质、蛋白质酶等发挥着重要作用。 目的：比较正常腰椎间盘组织及退变腰椎间盘组织中核因子κB、基质金属蛋白酶3的表达,分析两者的相关性。 方法：收集唐山市第二医院脊柱外科手术切除退变椎间盘组织68例(病变组),腰椎爆裂骨折正常椎间盘组织10例(对照组),应用苏木精-伊红染色、免疫组织化学方法和ELISA方法检测核因子κB、基质金属蛋白酶3的表达,并对两者相关性进行分析。 结果与结论：苏木精-伊红染色结果显示对照组可见纤维环和髓核结构,腰椎间盘髓核的软骨细胞表现为不规则的软骨陷窝,病变组细胞增殖明显,细胞核呈圆形或卵圆形,髓核细胞胞质呈空泡状。免疫组织化学方法显示病变组核因子κB、基质金属蛋白酶3吸光度值明显高于对照组(P < 0.05)。ELISA检测病变组核因子κB、基质金属蛋白酶3的表达均高于对照组(P < 0.05)。两者具有正相关性(r=0.643 0,P < 0.01)。     

模拟人后路髓核摘除构建椎间盘退行性变动物模型

背景：髓核摘除术是治疗椎间盘突出的经典方法,但存在较高的复发率。 目的：验证模拟人后路髓核摘除进行后外侧穿刺抽吸髓核,建立椎间盘退行性变动物模型的可行性。 设计、时间及地点：观察对照实验,于2006-10/2007-02在解放军南京军区福州总医院动物实验室完成。 材料：选用日本大耳白兔20只,行椎间盘退行性变动物模型制备。 方法：用持针器夹持21G穿刺针,行L1~2及L3~4椎间盘右后外侧穿刺髓核抽吸法摘除部分髓核组织,术后2,4,8,12周分别对造模后椎间盘行组织学观察,并将L2~3椎间盘作为对照。 主要观察指标：通过苏木精-伊红染色观察椎间盘组织学结构。 结果：苏木精-伊红染色可见对照椎间盘大多髓核完整,髓核与纤维环分界清晰,纤维环结构接近正常,髓核组织中有大量髓核细胞。造模后第4周髓核细胞数量不断减少,第12周时髓核中主要为纤维组织,伴有极少量髓核细胞。 结论：模拟人后路髓核摘除术建立后外侧纤维环穿刺髓核抽吸的椎间盘退行性变动物模型成功建立,可为应用组织工程修复重建退行性变椎间盘提供有效的动物模型。     

转化生长因子β1注射退变椎间盘内软骨终板的组织形态变化

背景：椎间失稳能导致软骨终板的退变,是椎间盘退变发病机制中的基础环节。 目的：观察退变椎间盘内注射转化生长因子β1后,椎间盘软骨终板的组织形态学变化。 方法：将日本大耳白兔随机分为对照组、预防组和治疗组。所有兔均建立L5~6,L6~7椎间失稳模型。预防组在完成椎间失稳建模后立即于损伤侧L5~6,L6~7椎间盘内注射转化生长因子β1,治疗组于椎间失稳建模后3个月行相同方法注射转化生长因子β1。 结果与结论：建模后3,6个月,预防组较对照组软骨终板软骨细胞分布均匀,潮线清晰。Mankin评分降低(P < 0.05)。建模后第6个月治疗组较对照组软骨终板表层光滑,细胞排列均匀,潮线清晰,染色均匀,Mankin评分降低(P < 0.05)。结果证实,在兔椎间盘内注射转化生长因子β1可延缓椎间盘软骨终板的退变。     

透明质酸钠溶液复合可注射型骨髓间充质干细胞修复退变椎间盘

摘要 背景：透明质酸作为椎间盘组织工程支架的基质材料,可提供蛋白多糖附着点,增加蛋白多糖的沉积,以足够大的孔率允许种子细胞长入。 目的：观察透明质酸钠混合溶液复合骨髓间充质干细胞修复兔退变椎间盘的效果。 方法：以后外侧穿刺抽吸L1/2和L3/4髓核构建日本大耳白兔椎间盘退变模型,造模后2周应用微量注射器向L3/4椎间盘内注射同种异体骨髓间充质干细胞与透明质酸钠混合溶液作为实验组,L1/2椎间盘注射透明质酸钠作为对照组。注射后2,4,8,12 周时行兔椎间盘影像学及病理学检查。 结果与结论：对照组椎间盘高度呈降低趋势,实验组注射后2周椎间盘高度缓慢下降,之后缓慢升高,两组在4个时间点椎间盘高度指数改变差异有显著性意义(P < 0.05)。病理结果显示实验组髓核纤维环形态得到保留,髓核纤维环边界清晰,植入的骨髓间充质干细胞产生增殖分裂,并向周围迁徙规律排列,其病理学评分明显高于对照组。12周内实验组Ⅱ型胶原含量高于对照组(P < 0.05)。说明移植于退变椎间盘内的骨髓间充质干细胞能够存活,且有增殖能力,其与透明质酸钠混合溶液联合移植可以延缓退变椎间盘进一步退变,并促进退变椎间盘修复。 关键词：透明质酸钠;骨髓间充质干细胞;椎间盘退变;细胞移植;髓核抽吸 doi:10.3969/j.issn.1673-8225.2011.16.004     

胰岛素样生长因子Ⅰ对大鼠纤维环细胞体外增殖活性影响的量效关系

背景：近年来体外椎间盘髓核和纤维环组织细胞培养技术的建立，特别是软骨组织工程研究的不断深入及自体椎间盘细胞移植修复髓核缺损动物实验的初步成功，为退变椎间盘的形态结构与生理功能的完全再生修复带来了希望。 目的：通过对椎间盘纤维环细胞的培养，观察胰岛素样生长因子Ⅰ对大鼠纤维环细胞体外增殖活性的影响及量效和时效关系。 设计、时间及地点：单一样本观察，于2006-09/2007-01在山西医科大学寄生虫实验室完成。 材料：清洁级1月龄 Wistar系大鼠30只，雌雄不限。 方法：体外分离培养大鼠纤维环细胞。剂量-效应实验：在培养的细胞中分别加入由含体积分数为0.01或0.1的小牛血清HAMF-12配制成的不同浓度的胰岛素样生长因子Ⅰ（0.1，1.0，10，100 μg/L）。以不加生长因子做对照，培养72 h。时间-效应实验：在培养的细胞中分别加入含有最佳效应浓度胰岛素样生长因子Ⅰ体积分数为0.1的小牛血清+F12培养液，分组为对照组和100 μg/L的胰岛素样生长因子Ⅰ组，分别培养1，3，5，7 d。 主要观察指标：①苏木精-伊红、甲苯胺蓝、免疫细胞化学染色分析细胞的生物学特性。②噻唑蓝比色法观察胰岛素样生长因子Ⅰ在体积分数为0.01和0.1的血清浓度下对大鼠纤维环细胞体外增殖的调节作用及其剂量、时间与作用效果的关系。 结果：苏木精-伊红染色细胞多为梭形，有伪足伸出，细胞核为圆形或椭圆形；甲苯胺蓝染色，胞浆为深蓝色；免疫细胞学方法检测表明纤维环细胞有Ⅰ型胶原表达；在体积分数为0.1的血清条件下，胰岛素样生长因子Ⅰ能提高细胞的增殖活性，并且在有效浓度范围内呈剂量效应关系。 结论：胰岛素样生长因子Ⅰ能促进大鼠纤维环细胞的体外增殖，其效应在一定范围内与剂量和时间呈正相关。     

Mechanism of action of tubocurarine on nicotinic cholinoreceptors of neurons of the sympathetic ganglia of rats

Tubocurarine (Tc) effect on membrane currents elicited by acetylcholine (ACh) was studied in isolated superior cervical ganglion neurons of rat using patch-clamp method in the whole-cell recording mode. The "use-dependent" block of ACh current by Tc was revealed in the experiments with ACh applications, indicating that Tc blocked the channels opened by ACh. Mean lifetime of Tc-open channel complex, tau, was found to be 9.8 +/- 0.5 s (n = 7) at -50 mV and 20-24 degrees C. tau exponentially increased with membrane hyperpolarization (e-fold change in tau corresponded to the membrane potential shift by 61 mV). Inhibition of the ACh-induced current by Tc (3-30 microM/1) was completely abolished by membrane depolarization to the level of 80-100 mV. Inhibition of ACh-induced current was augmented at increased ACh doses. It is concluded that the open channel block produced by Tc is likely to be the only mechanism for Tc action on nicotinic acetylcholine receptors in superior cervical ganglion neurons of rat.     

The effect of age of onset of PD on risk of dementia

Background Dementia occurs in the majority of patients with Parkinson’s disease (PD). Late onset of PD has been reported to be associated with a higher risk for dementia. However, age at onset (AAO) and age at baseline assessment are often correlated. The aim of this study was to explore whether AAO of PD symptoms is a risk factor for dementia independent of the general effect of age. Methods Two community-based studies of PD in New York (n = 281) and Rogaland county, Norway (n = 227) and two population-based groups of healthy elderly from New York (n = 180) and Odense, Denmark (n = 2414) were followed prospectively for 3–4 years and assessed for dementia according to DSM-IIIR. All PD and control cases underwent neurological examination and were followed with neurological and neuropsychological assessments. We used Cox proportional hazards regression based on three different time scales to explore the effect of AAO of PD on risk of dementia, adjusting for age at baseline and other demographic and clinical variables. Findings In both PD groups and in the pooled analyses, there was a significant effect of age at baseline assessment on the time to develop dementia, but there was no effect of AAO independent of age itself. Consistent with these results, there was no increased relative effect of age on the time to develop dementia in PD cases compared with controls. Interpretation This study shows that it is the general effect of age, rather than AAO that is associated with incident dementia in subjects with PD. Received in revised form: 22 December 2005     

Limits of deinstitutionalization--perspectives of relatives of psychiatric patients

After a hopeful beginning, the social process of the reintegration of those with severe mental illness has come to a standstill. I am led to wonder whether "the community" really wants to live together with people suffering from severe mental illness, and if so, how closely? As long as the medical treatment of mental illness provided by the general practitioners is fundamentally deficient, as they are not able to prescribe the necessary interventions--such as out-patient psychiatric nursing, and service providers in the out-patient sector are content with offering increasingly intensive forms of care for the less seriously ill at the cost of the Social Welfare System--the reintegration of those with serious mental illness remains an illusion--which is mainly to the benefit of providers of residential care in homes and hostels.     

Denervation study of synapses of organ of corti of old world monkeys

Fine structural characteristics of synapses in the spiral organ of Corti were examined, with reference to differences between inner and outer haircell systems, and to location of neurons of origin of efferent axons. Surgical interruption of crossed olivocochlear bundle, of vestibular nerve, of facial nerve, and excision of superior cervical ganglia were used to determine the pathways of efferent axons. Interruption of the vestibular nerve near the brainstem results in degeneration of all efferent terminals on outer hair cells. Mid-line lesions at, and caudal to, the facial colliculus result in degeneration of about half of these efferent terminals. Efferent synaptic bulbs to the inner hair-cell system are small, of the order of one micron, and form type 2 junctions with afferent dendrites. They tend to have more large dense-core vesicles (about 80 nm) than the large efferent terminals of the outer hair-cell system, and appear to be the terminals of axons in the habenula perforata, which exhibit varicosities laden with large dense core vesicles. The varicosities are unaffected by excision of the superior cervical ganglia. So far as our material can reveal, it appears that the varicosities in the habenula perforata do not survive vestibular root interruption, nor do the efferent processes in the internal spiral bundle or at the base of inner hair cells. Most interestingly, the afferent processes of the inner hair-cell system, as identified for example by their relation to pre-synaptic bodies in the inner hair cells, are subject to a trans-synaptic reaction after severance of the vestibular root. They undergo a dramatic cytological transformation, characterized by increase of volume, engorgement with microtubules, microfilaments, microvesicles of various sizes, and clusters of lysosomes. Thus, both the efferent and afferent terminals of the inner hair-cell system show marked cytological differences from the corresponding terminals of the outer hair cell system.     

Summary of legislation of 1935 of interest to the department of Mental Hygiene

Psychiatric Quarterly -