免疫反应在实验性游离型腰椎间盘突出自然吸收中的意义

目的：通过检测CD3和IgG在实验性游离型腰椎间盘突出自然吸收中的表达，探讨腰椎间盘突出后能否自然吸收及其可能的吸收机制。方法：将25只成年狗平均分成5组，从L2/3切取髓核组织，置于L5处的硬脊膜外，于术后2，4，6，8，12周，取动物标本以及正常部位的髓核组织，用CD3抗体和抗IgG进行免疫组化检测。结果：约30%的标本局部为脂肪组织，54%的标本局部硬脊膜与骨粘连，16%的标本既无粘连也无脂肪，免疫组化测定显示，随着时间的推移，CD3^ 细胞的浸润程度呈递减趋势，而IgG沉积的分布程度呈递增趋势，正常髓核无CD3^ 细胞浸润，IgG沉积分布极少。结论：在椎管内，狗游离的髓核组织8-12周后可部分或全部自然吸收，其吸收机制是机体自身免疫反应的结果。     

The development of fibrocartilage in the rat intervertebral disc

The development of fibrocartilage in rat lumbar intervertebral discs has been correlated with an immunohistochemical analysis of the changing distribution of extracellular matrix components. Disc anlagen were first recognised by embryonic day 14 as segmental cell condensations. By E16, the notochord formed a series of bulges, each representing a future nucleus pulposus, and the annulus fibrosus had differentiated in the disc anlagen. The inner part of the annulus was composed of cartilage which linked that of adjacent vertebral bodies. The outer part was fibroblastic, with layers of parallel fibroblasts. The long axes of the cells in successive layers lay at an angle of approximately 90° to each other. This criss-cross orientation of cells preceded the oriented deposition of collagen fibres to form the lamellae. Disc anlagen were immunolabelled weakly for types I and III collagen, chondroitin 6-sulphate and dermatan sulphate. Later tissue differentiation was marked by the appearance of type II collagen, chondroitin 4-sulphate and keratan sulphate in the inner annulus. These components also appeared in the outer annulus, but only in adult animals, and indicated metaplastic change in the lamellar fibroblasts. Fibrocartilage in the nucleus pulposus was only seen in old animals, and the origin of the tissue was less clear. However, the fibrocartilage cells appeared to be derived from the cartilage end plate and/or from the inner annulus. We conclude that fibrocartilage in the intervertebral disc is derived from several sources and that the radial distribution patterns of extracellular matrix components in the adult disc are explained by the embryonic origins of its parts.     

长链非编码RNA KLHL7-DT对人退变髓核细胞增殖和凋亡的影响及其相关机制

目的 探讨长链非编码RNA KLHL7-DT对人退变髓核细胞增殖和凋亡的影响及其相关机制。方法 选取2018年1月—2019年10月南京江北医院骨科脊柱骨折患者手术切除的正常椎间盘标本18例,其中男11例、女7例,年龄为22~46 （38.3±4.3）岁,PfirrmannⅠ级6例、Ⅱ级12例。取椎间盘标本常规分离、培养髓核细胞,使用10 ng/mL IL-1β处理髓核细胞获得退变髓核细胞。将退变髓核细胞分为沉默对照组、KLHL7-DT沉默组、过表达对照组、KLHL7-DT过表达组。4组细胞分别对应转染沉默对照序列、siRNA-KLHL7-DT沉默序列、过表达对照序列、KLHL7-DT过表达序列。取转染后4组退变髓核细胞采用5-乙炔基-2’脱氧尿嘧啶核苷（EdU）法检测细胞增殖情况,流式细胞术检测细胞凋亡情况,Western blot检测聚集蛋白聚糖（Aggrecan）、Ⅱ型胶原（Col Ⅱ）蛋白的表达情况。结果 （1）KLHL7-DT过表达组EdU染色阳性细胞/DAPI染色阳性比值（0.147±0.002）低于过表达对照组（0.203±0.007）,而KLHL7-DT沉默组比值（0.428±0.050）高于沉默对照组（0.240±0.032）,差异均有统计学意义（t=14.25、-5.44,P值均<0.05）。（2）KLHL7-DT过表达组细胞凋亡率（19.01%±0.41%）高于过表达对照组（14.38%±0.31%）,KLHL7-DT沉默组细胞凋亡率（16.08%±0.59%）低于沉默对照组（17.42%±0.36%）,差异均有统计学意义（t=15.69、3.36,P值均<0.05）。（3）Western blot结果显示,KLHL7-DT过表达组细胞Aggrecan和Col Ⅱ蛋白的相对表达量（分别为0.34±0.29、0.57±0.11）均低于过表达对照组（1.00±0.22、1.05±0.10）,KLHL7-DT沉默组Aggrecan和Col Ⅱ蛋白的相对表达量（分别为1.77±0.14、1.63±0.12）均高于沉默对照组（1.10±0.18、0.98±0.08）,差异均有统计学意义（t=3.10、5.54、-5.05、-7.66,P值均<0.05）。结论 上调KLHL7-DT的表达可抑制退变髓核细胞的增殖,促进退变髓核细胞的凋亡,其机制可能是通过调节细胞外基质Aggrecan、Col Ⅱ蛋白的合成,进而参与椎间盘退变的发展。     

八珍汤对腰椎间盘突出症微创孔镜下椎间盘髓核摘除患者的疼痛及生活质量的影响

目的:探究八珍汤对腰椎间盘突出症微创孔镜下椎间盘髓核摘除患者的疼痛及生活质量的影响。方法:选取余姚市人民医院脊柱外科 2019年 2月—2020年 2月选取腰椎间盘突出症微创孔镜下椎间盘髓核摘除患者,将患者随机分成两组,每组 65例,对照组行常规治疗,观察组在对照组的基础上加用八珍汤治疗。观察比较治疗 1个月后两组患者的疼痛情况、脊柱功能障碍情况,以及生活质量。结果:治疗前,患者视觉模拟(VAS)评分比较差异无统计学意义;治疗后,两组 VAS评分均低于治疗前,且观察组低于对照组,差异有统计学意义( P<0.05)。治疗前,患者脊柱功能障碍( ODI)评分比较差异无统计学意义;治疗后,两组 VAS评分均低于治疗前,但两组差异无统计学意义( P>0.05)。治疗后,观察组患者生活质量评分高于对照组,差异有统计学意义(P<0.05)。结论:腰椎间盘突出症微创孔镜下椎间盘髓核摘除患者加用八珍汤治疗,能够有效缓解疼痛,提升生活质量。     

脂肪间充质干细胞向髓核样细胞定向分化研究

目的探讨SD大鼠来源的髓核(nucleus pulposus,NP)细胞促使脂肪间充质干细胞(adipose tissue-derived mesenchymal stem cells,AMSCs)向NP样细胞定向分化的分子机制。方法采用酶消化法取脂肪细胞,极限稀释法纯化细胞;采用组织块培养法培养NP细胞。利用流式细胞技术,免疫荧光及RT-PCR检测对AMSCs及NP细胞进行鉴定。结果 AMSCs中Sca-1和CD44的阳性率较高,而CD45和CD11b阴性,共培养组荧光强度明显亮于单纯AMSCs组,AMSCs在NP细胞的诱导下聚焦蛋白聚糖(Aggrecan)、Ⅱ型胶原蛋白(CollagenⅡ)、Sox-9等表达水平较对照组高。结论共培养环境中髓核细胞分泌的可溶性因子TGF-1能促使AMSCs向NP样细胞定向分化。     

Antenatal differentiation of the human intervertebral disc

Summary To study the antenatal differentiation of the human intervertebral disc, the columns of forty eight embryos and fetuses were examined histologically. The primitive disc is composed of two structures: the notochord which shows a progressive expansion into the disc, and the fibrocartilaginous perinotochordal disc. No histological sign of interaction between notochordal and perinotochordal cells, which may explain the notochordal expansion into the discs, was seen. On the other hand, the notochordal intervention in the cartilaginous differentiation of the inner zone is probable.

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Différenciation anténatale du disque intervertébral humain

Résumé Cette étude de la différenciation anténatale du disque intervertébral humain repose sur l'examen de coupes histologiques de quarante huit colonnes vertébrales d'embryons et de foetus. Le disque primitif est composé de deux structures : la notochorde, qui présente une expansion progressive de son diamètre au sein du disque, et le disque périnotochordal, d'abord mésenchymateux puis fibrocartilagineux. Il n'a pas été mis en évidence de signe histologique témoignant d'une interaction entre les cellules notochordales et les cellules périnotochordales qui puisse expliquer l'expansion de la notochorde au sein des disques. Le rôle de la notochorde dans la différenciation cartilagineuse de la zone centrale est par contre probable.

     

Does a herniated nucleus pulposus contribute significantly to a decrease in height of the intervertebral disc?

A lumbar intervertebral disc with a herniated nucleus pulposus (HNP) often exhibits a decrease in the height of the intervertebral space. Our purpose was to ascertain whether the loss of volume of an HNP is sufficient to cause a perceptible decrease in the height of the intervertebral space. MRI of 44 patients with 51 HNPs were reviewed. The volumes of the herniated material and of the intervertebral discs were calculated for every level from L 1–2 to L 5–S 1. The average volume of the HNP was 503 ± 301 mm³. The average volumes of all 220 intervertebral discs and of the 127 normal-appearing discs were 14 442 ± 4200 mm³ and 17 476 ± 2885 mm³ respectively. The average volume of the HNP represented 3.5 % of the parent disc. An average HNP caused a decrease in intervertebral space height of 0.35 mm (0.56 pixels). Therefore, the loss of the volume of the HNP does not cause a significant decrease in the intervertebral space height. The average calculated decrease in the disc height is less than that reported in normal diurnal variation. Received: 1 February 1999/Accepted: 24 October 1999     

椎板开窗手术治疗腰椎间盘突出症108例分析

目的　探讨椎板开窗法摘除髓核治疗腰椎间盘突出症的可行性及其手术操作要点。方法　对10 8例腰椎间盘突出症行椎板开窗法摘除髓核术并对疗效作出评价。结果　采用椎板开窗法摘除髓核术治疗腰椎间盘突出症 ,疗效优良达 97%。结论　本术式具有操作简便、损伤小、出血少、病人恢复快等优点 ,值得临床推广。     

木瓜凝乳蛋白酶的分离纯化及体外溶核作用

目的:建立简单有效的木瓜凝乳蛋白酶分离纯化方法,并研究其体外溶核作用.方法:采用离子交换层析直接将木瓜凝乳蛋白酶分离纯化,体外试管法测定该酶的溶核作用.结果:建立了简单有效的木瓜凝乳蛋白酶分离纯化方法,其纯化产品能溶解人体椎间盘的髓核.结论:直接用离子交换层析柱来分离纯化木瓜凝乳蛋白酶是可行的,其体内溶核作用尚值得研究.     

GDF-5基因重组腺病毒修复人退变髓核细胞的实验研究

【摘要】 目的 探讨腺病毒介导的GDF 5基因对人退变髓核细胞外基质表达的影响,探求一种基因治疗新途径。方法 利用腺病毒载体将GDF 5基因导入退变髓核细胞,设置为空白对照组（Control组）、阴性对照组（GFP组）、实验组（GDF 5组）3组,并分为3、7、14、21天四个观察时间点,分别检测3组髓核细胞sGAG、Hyp的含量,免疫染色、Safranine O染色观察细胞外基质合成情况,Real time PCR检测Aggrecan、Collagen II mRNA表达水平。结果 GDF 5组髓核细胞sGAG/DNA、Hyp/DNA均在14、21天较Control组和GFP组明显增加,差异有统计学意义（P＜0.05）,而Control组与GFP组之间任意观察时间点比较差异均无统计学意义（P＞0.05）;免疫染色、Safranine O染色结果显示GDF 5组髓核细胞的蛋白多糖、Ⅱ型胶原、蛋白聚糖在14、21天均呈强阳性染色,Control组和GFP组呈弱阳性染色。Real time PCR结果显示GDF 5组Aggrecan、Collagen II mRNA的表达在14、21天较Control组和GFP组明显增加,差异有统计学意义（P＜0.05）。结论 腺病毒介导的GDF 5能明显促进人退变髓核细胞Aggrecan、Collagen II mRNA的表达,并增加蛋白多糖、Ⅱ型胶原的合成,对细胞外基质具有明确的修复作用,是一种有效的基因治疗新方法。