uPA、MMP-3在退变椎间盘中的表达及意义

目的探讨尿激酶型纤溶酶原激活剂（uPA）、基质金属蛋白-3（MMP-3）在人类退变椎间盘组织中的表达变化。方法对照组为取自脊柱侧凸患者的正常腰椎间盘组织；实验组为腰椎间盘突出患者的退变椎间盘组织。分别进行免疫组织化学染色，比较2组椎间盘组织中uPA及MMP-3的光密度值。结果uPA和MMP-3在突出椎间盘组织中平均光密度值较正常椎间盘组织中的平均光密度值表达明显升高；相关分析显示突出椎间盘中uPA、MMP-3呈正相关关系。结论uPA和MMP-3均可能参与了人椎间盘组织的退变过程，且MMP-3与uPA在人类退变椎间盘组织中有相同的增高趋势。     

退变腰椎间盘组织中碱性成纤维细胞生长因子的表达研究

目的 探讨碱性成纤维细胞生长因子（basic fibroblast growth factor,bFGF)在正常和退变椎间盘组织中的表达情况。方法 交30例来源于腰椎间盘突出症患者手术中所取得的椎间盘组织（观察组，男11例，女19例；年龄25－78岁，平均48岁；病程3个月－30年，平均9年11个月）与6例来源于脊柱侧凸患者前路松解术所取得的椎间盘组织（对照组，男女各3例，年龄10－17岁，平均14.2岁）进行对比，首先经病理组织学检查证实为退变椎间盘组织和政党椎间盘组织，然后将两组椎间盘组织分别通过免疫组织化学方法和原位杂交方法，检测各自椎间盘组织中的bFGF及其mRNA的表达。观察组30例均为退变椎间盘组织，免疫组化阳性率为90％（27／30），原位杂交阳性率为20％（6／30）；对照组6例均为正常椎间盘组织，其免疫组及原位杂交均为阴性，两组间免疫组化方法检测阳性率在统计学上差异有非常显著性意义。结论 bFGF在正常和退变椎间盘组织中表达的差异有显著性意义。提示 bFGF可能作为增生刺激因子促进椎间盘组织中的软骨细胞增生和细胞外基质合成，进而加速椎间盘退变。     

MMP-3、IgG和CD68在青少年与中老年突出腰椎间盘组织中的表达分析

【摘要】 目的：比较细胞因子基质金属蛋白酶3（matrix metalloproteinases 3,MMP-3）、IgG和CD68在青少年与中老年患者突出腰椎间盘组织中的表达,探讨两个年龄段腰椎间盘突出的病因。方法：收集40例腰椎间盘突出症患者的突出腰椎间盘髓核标本,青少年组18例,年龄11～25岁,平均20.6±3.4岁;中老年组22例,年龄40～72岁,平均48.0±10.3岁。HE染色观察标本退变情况,免疫组化染色检测MMP-3、IgG和CD68的表达,光学显微镜下观测并计录数据。结果：标本HE染色提示多数（17/22）中老年组腰椎间盘存在明显退变,青少年组多数椎间盘（13/18）退变不明显或无退变。MMP-3阳性率青少年组（33.33%）低于中老年组（81.82%）;IgG阳性率青少年组（66.67%）高于中老年组（31.82%）;CD-68阳性率青少年组（83.33%）高于中老年组（45.45%）,两组MMP-3、IgG和CD68的表达阳性率比较,差异均有统计学意义（P<0.05）。结论：免疫和炎症反应可能是青少年腰椎间盘突出的重要病因,而中老年腰椎间盘突出可能主要与退变有关。     

新型山羊腰椎间盘突出模型的建立

目的建立新型的山羊腰椎间盘退变合并突出的动物模型。方法9只6月龄山羊，随机分成3组，每组18个腰椎间盘，再将每只山羊的6个腰椎间盘随机等分成正常组和模型组。模型组椎间盘左前外侧纤维环用直径2．5mm的微型环锯限深5mm造成柱状纤维环缺损。术后2、4、6个月时对正常及模型椎间盘摄X线片和MR扫描，观察X线片上的椎间盘高度指数、MRI T2加权像上的椎间盘高信号区的面积和平均R弛豫时间、椎间盘纤维环损伤区的形态学变化，进行组织病理学切片观察（HE染色、Masson三色胶原染色和番红O-固绿染色），对椎间盘组织形态学变化、生物化学变化和组织学评分等指标进行评估。结果正常成年山羊椎间盘组织学结构特点与人类相似；模型椎间盘在损伤后2个月即出现明显退变（P〈0．01），包括椎间盘高度指数降低、椎间盘T2加权像高信号区面积和平均R弛豫时间减低、组织学退变评分增高；MRI和组织学均发现损伤纤维环外侧1／3修复，内2／3出现髓核组织突出充填。损伤后4、6个月模型椎间盘退变加重，但与损伤后2个月的指标差异无统计学意义（P〉0．05）；椎间盘突出形态稳定。结论以成年山羊为对象，应用微型环锯对椎间盘前外侧纤维环定量损伤可以获得快速、有效、重复性好的腰椎间盘突出动物模型。     

TNF-α、IL-1β在退变椎间盘组织中的表达及其意义

目的探讨肿瘤坏死因子-α（TNF—α）、白细胞介素-1β（IL-1β）在人类突出椎间盘组织中的表达变化及意义。方法实验组为手术切除的腰椎间盘突出患者的退变椎间盘组织,分为纤维环破裂组和未破裂组;对照组为取自腰椎骨折患者的正常腰椎间盘组织。采用双抗体夹心ABC—ELISA法分别测定3组椎间盘组织中的TNF-α、IL-1β的含量并进行比较。结果TNF-α和IL-1β含量在纤维环破裂组和纤维环未破裂组均明显高于对照组,纤维环破裂组明显高于纤维环未破裂组（P〈0．05）。结论TNF—α和IL-1β均可能参与了人类椎间盘组织的退变过程,且两者在突出的椎间盘组织中有相同的增高趋势。     

退变腰椎间盘组织的血管浸润现象及其意义

目的：观察退变腰椎间盘的血管浸润现象。方法：对５７例经手术治疗的腰椎间盘突出症患者的退变椎间盘组织及３９个（１３例）正常椎间盘组织做了组织学观察。结果：破裂型突出的血管浸润率（７２．０％）高于凸起型（１８．８％）；病程在一年以内的血管浸润率（５１．２％）高于病程超过一年者（１８．８％），患者年龄与血管浸润无直接关系。结论：腰椎间盘突出的不同病理类型不仅是形态学上的差异，也具有各自不同的组织学基础，破裂型突出的较高血管浸润率很可能与其较严重的临床表现以及较明显的组织退变相关联。血管浸润主要出现在腰椎间盘突出的早期或急性阶段，这不仅是该病变的一种伴随现象，很可能在椎间盘退变中起关键性的始动作用     

环氧化酶2与血管内皮生长因子在突出腰椎间盘中的表达及意义

目的：探讨环氧化酶2（COX-2）、血管内皮生长因子（VEGF）在突出腰椎间盘中的表达及其意义。方法：62个突出椎间盘标本取自58例腰椎间盘突出症手术患者,包括突起型22个,破裂型20个,游离型20个。取材部位分别为突出组织或游离组织（A部位）和椎间隙残余髓核组织（B部位）。对照组取自4例新鲜年轻尸体的L3/4、L4/5及L5/S1椎间盘组织共12个标本,取材部位为椎间盘边缘（A部位）和中央髓核（B部位）;应用免疫组化法对各组标本中COX-2和VEGF的表达进行检测;应用图像分析系统测量标本中COX-2和VEGF表达量的平均光密度值。结果：在腰椎间盘突出组,特别是破裂型和游离型组的突出组织中存在富含新生血管的肉芽组织,COX-2和VEGF染色阳性细胞主要表达于肉芽组织中及突出组织的椎间盘细胞中,对照组未见阳性染色细胞。在突出组的A部位从突起型、破裂型到游离型COX-2和VEGF的表达均逐渐增高,差异有显著性（P〈0.01）。突出组的A部位COX-2和VEGF的表达明显高于B部位（P〈0.01）。腰椎间盘组织中COX-2和VEGF的表达存在明显相关性（r=0.855,P〈0.01）。结论：COX-2、VEGF参与腰椎间盘退变、突出的发病过程;随着腰椎间盘退变突出的进展,COX-2、VEGF的表达均逐渐增高;COX-2与VEGF表达密切相关。     

青少年与中老年腰椎间盘突出相关因素分析

[目的]检测细胞因子基质金属蛋白酶(matrix metalloproteinases,MMPs)MMP-3,IgG和CD68在青少年与中老年患者突出腰椎间盘组织中的表达,探讨两个年龄段腰椎间盘突出的病因.[方法]收集42例腰椎间盘突出症患者的突出腰椎间盘髓核标本,青少年组18例;年龄11 ～25岁,平均(20.59 ±3.39)岁;中老年组24例,年龄30～72岁,平均(47±11.15)岁.HE染色观察标本退变情况,免疫组化检测MMP-3,IgG和CD68的表达,光学显微镜下观测并记录数据.[结果] MMP-3阳性率青少年组(33.3％)低于中老年组(79.2％);IgG阳性率青少年组(66.7％)高于中老年组(29.2％);CD-68阳性率青少年组(83.3％)高于中老年组(54.2％),两组MMP-3,IgG和CD68的表达阳性率比较,均P＜0.05,差异均有统计学意义.标本HE染色提示中老年组腰椎间盘存在明显退变,青少年组椎间盘退变不明显或无退变.[结论]病理表明中老年组突出腰椎间盘组织存在明显退变,而青少年组退变不明显.免疫组化检测显示中老年组MMP-3明显高于青少年组,但青少年组IgG和CD68均明显高于中老年组,免疫和炎症反应可能是青少年腰椎间盘突出的重要病因,而中老年腰椎间盘突出可能主要与退变有关.     

动物模型椎间盘退变全程基因变化的对比

目的：新西兰大白兔的纤维环损伤制作腰椎间盘退变模型，用以证实和比较在人椎间盘退变中明显变化的基因变化情况。方法：2．0只新西兰大白兔L4，5及L5,6纤维环损伤作为实验组，4只作对照组未行损伤。2、4、8、40周分别行核磁共振及计算机扫描摄影拍片证实椎间盘退变情况同时取椎间盘行精确定量逆转录聚合酶链式反应检测金属蛋白酶(matrix metalloproteinase)MMP-1、MMP-2、MMP-3及它们的特意性抑制剂(tissue inhibitors of metalloproteinases)Timp-1、Timp-2的变化。结果：核磁共振及计算机扫描摄影证实了腰椎纤维环损伤后腰椎逐渐退变并与人类退变结果相似，基因表达情况MMP-1、MMP-2、MMP-3、Timp-1、Timp-2早期均上调，MMP-3、Timp-1在退变的晚期出现下调。结论：纤维环损伤可以成功制作出椎间盘退变的模型。在人类腰椎间盘退变中明显上调的基因在此退变模型椎间盘中被发现同样上调，从而在分子水平证实了动物退变模型与人类的相似性。     

凋亡相关基因产物Fas APO-1蛋白在腰椎间盘组织中的表达及其意义

我们采用免疫组织化学方法对 35例腰椎间盘突出症患者的退变腰椎间盘及 11个 (7例 )正常腰椎间盘组织中ＦａｓＡＰＯ 1蛋白表达进行检测 ,探讨其在腰椎间盘退变及突出症发生、发展中的作用。一、材料与方法1.标本来源 :退变腰椎间盘组织(Ｌ1～ 5)取自本院骨科 1998年 5月～1999年 1月间 35例经手术治疗的腰椎间盘突出症患者 ,其中男 2 0例 ,女 15例 ,年龄 2 0～ 6 9岁 ,平均 40 .0岁。术中取纤维环及髓核组织各 1块 ,标本经病理学检查证实为退变椎间盘。 11个 (7例 )正常椎间盘组织 (Ｌ1～ 5)做为对照 ,标本取自新鲜尸检 ,年龄 2 2～ 5 9岁 …     

Bone morphogenic protein-2 signaling in human disc degeneration and correlation to the Pfirrmann MRI grading system

BACKGROUND CONTEXTBack and neck pain secondary to disc degeneration is a major public health burden. There is a need for therapeutic treatments to restore intervertebral disc (IVD) composition and function.PURPOSETo quantify ALK3, BMP-2, pSMAD1/5/8 and MMP-13 expression in IVD specimens collected from patients undergoing surgery for disc degeneration, to correlate ALK3, BMP-2, pSMAD1/5/8 and MMP-13 expression in IVD specimens to the 5-level Pfirrmann MRI grading system, and to compare ALK3, BMP-2, pSMAD1/5/8 and MMP-13 expression between cervical and lumbar degenerative disc specimens.STUDY DESIGNAn immunohistochemical study assessing ALK3, BMP-2, pSMAD1/5/8, and MMP-13 expression levels in human control and degenerative IVD specimens.METHODSHuman IVD specimens were collected from surgical patients who underwent discectomy and interbody fusion at our institution between 1/2015 and 8/2017. Each patient underwent MRI prior to surgery. The degree of disc degeneration was measured according to the 5-level Pfirrmann MRI grading system. Patients were categorized into either the 1) control group (Pfirrmann grades I-II) or 2) degenerative group (Pfirrmann grades III-V). Histology slides of the collected IVD specimens were prepared and immunohistochemical staining was performed to assess ALK3, BMP-2, pSMAD1/5/8, and MMP-13 expression levels in the control and degenerative specimens. Expression levels were also correlated to the Pfirrmann criteria. Lastly, the degenerative specimens were stratified according to their vertebral level and expression levels between the degenerative lumbar and cervical discs were compared.RESULTSFifty-two patients were enrolled; however, 2 control and 2 degenerative patients were excluded due to incomplete data sets. Of the remaining 48 patients, there were 12 control and 36 degenerative specimens. Degenerative specimens had increased expression levels of BMP-2 (p=.0006) and pSMAD1/5/8 (p<.0001). Pfirrmann grade 3 (p=.0365) and grade 4 (p=.0008) discs had significantly higher BMP-2 expression as compared to grade 2 discs. Pfirrmann grade 4 discs had higher pSMAD1/5/8 expression as compared to grade 2 discs (p<.0001). There were no differences in ALK3 or MMP-13 expression between the control and degenerative discs (p>.05). Stratifying the degenerative specimens according to their vertebral level showed no significant differences in expression levels between the lumbar and cervical discs (p>.05).CONCLUSIONSBMP-2 and pSMAD1/5/8 signaling activity was significantly upregulated in the human degenerative specimens, while ALK3 and MMP-13 expression were not significantly changed. The expression levels of BMP-2 and pSMAD1/5/8 correlate positively with the degree of disc degeneration measured according to the Pfirrmann MRI grading system.CLINICAL SIGNIFICANCEBMP-SMAD signaling represents a promising therapeutic target to restore IVD composition and function in the setting of disc degeneration.     

P物质阳性神经纤维在退变腰椎间盘的分布及意义

目的研究P物质（substance P,SP）阳性神经纤维在腰椎间盘中的出现及分布,以探讨其在椎间盘退变中的潜在作用。方法收集16例患者的21个病变腰椎间盘和来自于新鲜尸体的12个正常对照椎间盘,行组织学检查和SP免疫组织化学染色检查。结果SP阳性神经纤维偶见于正常椎间盘,在病变椎间盘内可见较多的SP免疫反应阳性神经纤维分布,阳性神经纤维数量与腰椎间盘退变程度呈正相关。结论腰椎间盘退变程度和SP阳性神经纤维数量有明显的相关性,SP可能作为神经炎性介质加速了腰椎间盘的退变。     

重度肩锁关节脱位的手术治疗

目的：对15例重度肩锁关节脱位的手术7治疗进行治疗,方法：15例全部为Allman分型中的Ⅲ型损伤,其中有11例切除纤维软骨盘,7例修复锁韧带,3例加用一枚松质骨螺钉固定于锁骨与喙空间,肩锁韧带全部修复,用两枚克氏针交叉固定于肩锁关节。结果：经10个月－6年的随访。疗效评价按Karlsson分类A级为12例,B级3例,病人均重返原工作岗位,有10例喙锁间隙软组织钙化,但对病人肩关节活动并无影响,结论：对重度肩锁关工锐位的病人应尽早手术治疗。克氏针交叉内固定是一种简单而有效的方法,纤维软骨盘是否切除和喙锁韧带是否修复对预后无明显影响。     

NF-κB inhibitor,NEMO-binding domain peptide attenuates intervertebral disc degeneration

BACKGROUND CONTEXTNonphysiological mechanical loading and inflammation are both critically involved in intervertebral disc (IVD) degeneration, which is characterized by an increase in cytokines and matrix metalloproteases (MMPs) in the nucleus pulposus (NP). This process is known to be mediated by the NF-κB pathway.CLINICAL SIGNIFICANCECurrent clinical treatments for IVD degeneration focus on the alleviation of symptoms rather than targeting the underlying mechanism. Injection of an NF-κB inhibitor may attenuate the progression of IVD degeneration.PURPOSETo investigate the ability of the NF-κB inhibitor, NEMO binding domain peptide (NBD), to alter IVD degeneration processes by reducing IL-1β- and mechanically-induced cytokine and MMP levels in human nucleus pulposus cells in vitro, and by attenuating IVD degeneration in an in vivo rat model for disc degeneration.STUDY DESIGNExperimental in vitro and animal model.PATIENT SAMPLEDiscarded specimens of lumbar disc from 21 patients, and 12 Sprague Dawley rats.OUTCOME MEASURESGene and protein expression, cell viability, µMRI and histology.METHODSIL-1β-prestimulated human nucleus pulposus cells embedded into fibrin constructs were loaded in the Flexcell FX-5000 compression system at 5 kPa and 1 Hz for 48 hours in the presence and absence of NBD. Unloaded hNPC/fibrin constructs served as controls. Cell viability in loaded and unloaded constructs was quantified, and gene and protein expression levels determined. For in vivo testing, a rat needle disc puncture model was employed. Experimental groups included injured discs with and without NBD injection and uninjured controls. Levels of disc degeneration were determined via µMRI, qPCR and histology. Funding sources include $48,874 NASS Young Investigator Research Grant and $119,174 NIH 5K01AR071512-02. There were no applicable financial relationships or conflicts of interest.RESULTSMechanical compression of hNPC/fibrin constructs resulted in upregulation of MMP-3 and IL-8. Supplementation of media with 10 μM NBD during loading increased cell viability, and decreased MMP-3 gene and protein levels. IVD injury in rat resulted in an increase in MMP-3, IL-1β and IL-6 gene expression. Injections of 250 µg of NBD during disc injury resulted in decreased IL-6 gene expression. µMRI analysis demonstrated a reduction of disc hydration in response to disc needle injury, which was attenuated in NBD-treated IVDs. Histological evaluation showed NP and AF lesion in injured discs, which was attenuated by NBD injection.CONCLUSIONSThe results of this study show NBD peptide's capacity to reduce IL-1β- and loading-induced MMP-3 levels in hNPC/fibrin constructs while increasing the cells’ viability, and to attenuate IVD degeneration in rat, involving downregulation of IL-6. Therefore, NBD may be a potential therapeutic agent to treat IVD degeneration.     

腰椎椎间隙高度与上位椎体高度的比值与椎间盘退行性变Pfirrmann分级的相关性

目的 探讨腰椎椎间隙高度与上位椎体高度的比值与椎间盘退行性变程度之间的关系,为腰椎椎间盘退行性疾病的诊断和治疗提供客观准确的依据。方法 回顾性分析2019年1月—2019年6月来本院就诊的61例腰椎椎间盘退行性变患者临床资料。在腰椎侧位X线片上测量腰椎椎间隙及相应上位椎体的高度,并计算椎间隙高度与上位椎体高度的比值;在腰椎矢状位MRI上评估腰椎椎间盘退行性变Pfirrmann分级;比较不同Pfirrmann分级椎间盘的椎间隙高度与上位椎体高度比值的差异,并采用Spearman相关分析研究椎间隙高度与上位椎体高度比值与相应节段椎间盘Pfirrmann分级之间的相关性。结果 除L₁/L₂节段,其余各节段椎间隙高度与上位椎体高度比值均随着Pfirrmann分级增加而逐渐减小,差异均有统计学意义（P < 0.05）。相同Pfirrmann分级的不同节段椎间盘之间椎间隙高度与上位椎体高度比值差异无统计学意义（P > 0.05）。Spearman相关分析结果显示,L₂/L₃、L₃/L₄、L₄/L₅、L₅/S₁节段Pfirrmann分级与椎间隙高度与上位椎体高度比值呈负相关（r =-0.568,P < 0.05）。结论 临床上测量L₂/L₃、L₃/L₄、L₄/L₅、L₅/S₁节段椎间隙高度与上位椎体高度比值对腰椎椎间盘退行性疾病的诊断可能具有重要意义。     

Association and histological characteristics of endplate injury and intervertebral disc degeneration in a rat model

IntroductionThe purpose of this study was to construct a rat caudal vertebral body fracture model and to analyze the association and histological characteristics of vertebral body fracture with endplate injury and adjacent intervertebral disc degeneration.Materials and methodsThis study included 144 clean-grade male Sprague-Dawley rats, which were randomly divided into a control, middle vertebral body injury (MI), and endplate injury (EI) groups. A vertebral body fracture with or without endplate injury was developed by either drilling a hole in the middle of a rat caudal vertebral body to create a fracture with an intact endplate or drilling a hole in the vertebral body near the intervertebral disc to create a vertebral body fracture with endplate injury. The histological differences in the adjacent intervertebral discs of vertebral body fractures with or without endplate injury were detected using imaging, non-specific histological staining, immunohistochemistry and TUNEL assay.ResultsImaging results revealed that the EI group showed a significant decrease in intervertebral space height and intervertebral disc T2 signal over time. Non-specific histological staining revealed that in the EI group, the intervertebral disc was degenerative. Immunohistochemistry indicated that Aggrecan and Collagen-II were decreased and inflammatory factors were increased in the EI group. The TUNEL detection found that apoptosis was significantly increased in the EI group as compared with the MI and control groups.ConclusionIn rat caudal vertebral body fractures, a fracture with endplate injury is more likely to induce or accelerate degeneration of adjacent intervertebral discs.