纤维环切开法建立猪腰椎间盘退变动物模型

目的 建立猪腰椎间盘退变动物模型. 方法 6头实验猪全麻下行腰椎腹膜后入路,暴露椎间盘前外侧.18号穿刺针平行于椎间隙刺穿纤维环,深度17 mm,穿刺8个椎间隙.23号手术刀片平行于椎间隙刺穿纤维环,深度17mm,穿刺25个椎间隙.对照组9个椎间隙只暴露不损伤纤维环.术前及术后2周全部行腰椎MRI检查评估,术后第4、6周对其中特定1头继续进行MRI检查,并于6周时处死取椎间盘组织病理检查.结果 刀刺模型组术后2周出现明显的椎间盘退变,MRI T2加权像髓核相对信号强度从0.98±0.13降到0.77±0.17;椎间隙高度从(2.64±0.41)mm降到(1.98±0.32)mm;Pfirrmann分级从术前的Ⅰ级25例,变为术后Ⅱ级18例、Ⅲ级7例.刀刺模型组手术前后对比及术后与另外2组相比,上述指标差异显著(P＜0.05或P＜0.01).这种退变现象随着时间的推移有加重趋势.病理切片显示椎间盘髓核细胞减少,为纤维组织取代,纤维环板层裂隙明显,内层有毛细血管长入.针刺组除椎间盘高度有较小的降低外,余与对照组相似. 结论 刀刺损伤家猪椎间盘纤维环可以建立较理想的椎间盘退变动物模型,使用适当大小的针穿刺纤维环对椎间盘退变的影响不大.     

外源性IGF-1、OP-1对退变椎间盘的修复作用

目的为细胞因子治疗椎间盘退变提供理论支持。方法采用Lindblom折尾加压法建立鼠尾椎椎间盘退变模型,将制模成功后大鼠随机分为A、B、C、D组各15只,A、B、C组用微量泵分别注入外源性胰岛素样生长因子-1（IGF-1）、成骨蛋白-1（OP-1）及生理盐水,注射速度为5μl/min,注射完毕于进针处涂红霉素软膏,1周后于相同节段椎间盘不同的进针部位进行第2次注射,方法同第1次。D组不予任何处理。4周后取材,光镜下观察椎间盘形态、厚度,软骨陷窝中细胞密度,分析AB-PAS染色标本髓核中蛋白多糖的平均光密度（MOD）值。免疫组织化学SP法检测鼠尾椎椎间盘软骨板内细胞PCNA表达情况,计算阳性表达率。结果与C、D组比较,A、B组内层纤维环胶原明显增粗,排列整齐,陷窝中细胞密度明显增高;髓核完整性较好,无明显缺失;椎间盘厚度、髓核的MOD值均明显增加,B组软骨细胞PCNA阳性表达率均明显高于A、C、D组。结论鼠尾椎椎间盘内注射IGF-1、OP-1可刺激细胞增殖、增加胶原和蛋白多糖释放,促进退变椎间盘的修复;且OP-1效果优于IGF-1。     

手法治疗老年性腰椎间盘变性引起的坐骨神经痛

老年性椎间盘退变中,纤维环及髓核出现纤维性变,组织脆性增加,以至出现点片状钙化的纤维环碎片,在椎间盘内外力平衡失调的情况下,使纤维环碎片脱离附着点,引起神经根或周围韧带机械性刺激压迫症状。患者出现活动受限,咳嗽、打喷嚏时疼痛加     

经皮旋切术结合臭氧注射老年颈椎间盘突出的疗效

<正>颈椎间盘突出症属于临床上较为常见的颈椎病之一,常由于长时间疲劳损伤导致椎间盘出现进行性退变,髓核吸震能力由于含水量降低而减弱,当受到一些外伤等因素的影响下,导致纤维环破裂,髓核组织脱出,出现头晕、头痛、上肢麻木等临床症状,病情严重者可危及生命。本研究分析单纯经皮旋切术和经皮旋切术结合臭氧注射治疗颈椎间盘突出的治疗效果。     

腰椎间盘髓核及软骨终板切除的组织学及生物力学变化研究

将9只1岁健康犬随机分为A、B、C组,每组各3只。A组行假手术,B、C组建立腰椎间盘切除动物模型后分别行单纯髓核摘除术及髓核与软骨终板切除术,手术部位分别为L5-L6、L6～L7和L7～S1。术后分别于2、8、24周内处死动物,取出手术节段椎间盘,光镜、电镜下观察组织学变化。术后24周取出L5～S1椎体采用生物力学实验机测量相应节段在前屈、后伸、侧弯和轴向旋转载荷下的生物力学稳定性。结果B组术后手术节段椎间盘随着时间的延长出现明显退变,24周时大部分为坏死细胞;C组术后椎间隙内有大量纤维软骨形成,椎间盘内可见新生血管,24周时退变程度较轻。B组脊柱节段稳定性差,屈／伸方向上施加载荷时,运动范围（ROM）和中性区（NZ）均明显增大,在侧弯和轴向旋转方向上施加载荷时,NZ明显增大,与A组比较有显著差异;C组脊柱节段在各方向上的ROM和NZ与A组比较无显著性差异。认为髓核及软骨终板同时切除可促进纤维软骨生成及血管再生,保持脊柱节段的生物力学稳定性,防止术后腰椎不稳的发生。     

退变对腰椎4～5节段椎间盘生物力学的影响

目的 建立正常人体腰椎4～5节段(L4～5)有限元模型,基于正常模型建立退变椎间盘有限元模型,分析退变后L4～5椎间盘生物力学的变化.方法 选择健康男性志愿者1名,采集其正常腰椎CT和MRI的影像数据,建立正常L4～5有限元模型;通过改变纤维环和髓核的材料属性,建立退变L4～5椎间盘的有限元模型.设定约束条件,并在L4...     

退变腰椎间盘组织中p-JNK的表达变化及意义

目的 观察退变腰椎间盘组织中p-c-Jun氨基末端激酶(JNK)的表达变化,并探讨其意义.方法 选择手术切除的腰椎间盘突出症退变腰椎间盘组织(观察组,50例份)及腰椎爆裂骨折、椎间盘突出症正常腰椎间盘组织(对照组,20例),光学显微镜下观察两组腰椎间盘组织病理变化,免疫荧光法检测p-JNK的定位,Western blot法检测p-JNK蛋白.结果 光镜下,观察组细胞增殖明显,细胞核呈圆形或卵圆形,髓核细胞细胞质成空泡状,基质增多,纤维网错乱,纤维排列紊乱;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,细胞核呈卵圆形,蓝染,细胞质粉染,基质较均匀,可见排列整齐的纤维网.免疫荧光示,观察组细胞质和细胞核内均有p-JNK表达,对照组细胞质及细胞核内仅少量表达.Western blot法示,与对照组比较,观察组p-JNK蛋白水平升高(P＜0.05).结论 p-JNK在退变椎间盘组织中的表达明显高于正常椎间盘,提示p-JNK可能与椎间盘退变有关.     

不同前路融合固定手术治疗老年颈椎间盘突出症的疗效

颈椎间盘突出症主要是由于颈椎间盘髓核、纤维环、软骨板,尤其是髓核,发生不同程度的退行性病变后,在外界因素的作用下,导致椎间盘纤维环破裂,髓核组织从破裂之处突出或脱出椎管内,从而造成相邻的组织,如脊神经根和脊髓受压,引起头痛、眩晕、心悸、胸闷、颈部酸胀、活动受限、肩背部疼痛、上     

糖尿病大鼠椎间盘退变的实验观察

目的 观察糖尿病大鼠椎间盘的退变情况,探讨糖尿病引起椎间盘退变的可能机制.方法 30只雄性SD大鼠随机分为实验组和对照组,每组15只.实验组采用腹腔单次注射链脲佐菌素溶液(STZ)50 mg/kg制成糖尿病模型,正常对照组仅注射等量的枸橼酸缓冲液.分别在建模成功后4、8、12 w取椎间盘组织,HE染色观察椎间盘组织病理变化;透射电镜观察髓核细胞及胶原变化;用TUNEL法显示髓核组织细胞凋亡情况,测定凋亡率. 结果 实验组大鼠注射STZ后,空腹血糖均大于13.8 mmol/L[(20.27±2.600)mmol/L].在不同的阶段,实验组椎间盘的退变程度均较重,髓核细胞凋亡率较高.结论 糖尿病是影响椎间盘退变的因素之一.     

AKT-mTOR信号通路在大鼠椎间盘软骨终板细胞自然退变中的意义

目的探讨AKT-雷帕霉素靶蛋白(mTOR)信号通路在大鼠椎间盘软骨终板细胞自然退变中的表达情况。方法取清洁级SD大鼠12只,分离并培养椎间盘软骨终板细胞,建立体外自然传代退变模型。随机分为空白对照组(自然传代第2代细胞),自然退变组(自然传代第5代细胞)、AKT-mTOR信号通路抑制组(含LY294002抑制剂培养基传代至第5代),AKT-mTOR信号通路激动组(含IGF-1激动剂培养基传代至第5代)。倒置显微镜下观察各组细胞形态;甲苯胺蓝染色检测细胞蛋白聚糖变化;RT-PCR检测细胞中Ⅱ型胶原、蛋白多糖、SOX9以及AKT-mTOR信号通路中关键基因mTOR的表达情况;Western印迹检测AKT、p-AKT蛋白表达情况。结果随着传代次数增加,大鼠软骨终板细胞的表现逐渐减少。自然退变组Ⅱ型胶原、蛋白多糖、SOX9、mTOR的表达水平均明显低于空白对照组(P0.05),AKT-mTOR信号通路抑制组表达水平明显低于自然退变组(P0.05);AKT-mTOR信号通路激动组表达水平明显高于自然退变组(P0.05)。四组的AKT蛋白表达水平无统计学差异(P0.05);抑制组p-AKT蛋白表达水平低于另外3组,而激动组p-AKT蛋白表达水平明显高于抑制组和自然退变组(P0.05)。结论 AKT-mTOR信号通路在大鼠椎间盘软骨终板细胞自然退变中具有重要作用,添加AKT-mTOR信号通路抑制剂、激动剂能够加速、抑制细胞体外退变的发生。     

A preliminary study of neck-stomach syndrome

AIM: To determine the expression of c-Fos, caspase-3 and interleukin-1β (IL-1β) in the cervical cord and stomach of rats with cervical spondylosis, to analyze their relationship, and to offer an explanation of one possible cause for functional dyspepsia (FD) and irritable bowel syndrome (IBS) caused by cervical spondylosis. METHODS: The cervical spondylosis model in rats was established by destroying the stability of cervical posterior column. The cord segments C4-6 and gastric antrum were collected 3 mo and 5 mo after the operation. Rats with the sham operation were used as controls. The expressions of c-Fos, caspase-3 and IL-1β in the cervical cord and gastric antrum were determined by immunohistochemistry and/or Western blot. RESULTS: Immunohistochemical staining showed a few c-Fos, caspase-3 and IL-1β-positive cells in the cervical cord and antrum in the control. There was a significant increase in c-Fos, caspase-3 and IL-1β expression in model groups compared to the control groups at 3 mo and 5 mo after operation. More importantly, there was a significant (P < 0.05) increase in c-Fos, caspase-3 and IL-1β expression in the model group rats at 3 mo compared to those at 5 mo after the operation (c-Fos: 11.20 ± 2.26 vs 27.68 ± 4.36 in the cervical cord, 11.3 ± 2.3 vs 29.3 ± 4.6 in the gastric antrum; caspase-3: 33.83 ± 3.71 vs 36.32 ± 4.01 in the cervical cord, 13.23 ± 3.21 vs 26.32 ± 4.01 in the gastric antrum; IL-1β: 42.06 ± 2.95 vs 45.91 ± 3.98 in the cervical cord, 26.56 ± 2.65 vs 32.01 ± 2.98 in the gastric antrum). Western blotanalysis showed time-dependent changes of caspase-3 and IL-1β protein in the cervical cord and gastric antrum of rats with cervical spondylosis; there was no significant expression of caspase-3 and IL-1β protein in the control group at 3 mo and 5 mo after the sham operation, whereas there was a significant difference in caspase-3 and IL-1β protein levels between the model group rats followed up for 3 mo and those for 5 mo (P < 0.05). CONCLUSION: There is a significant association of c-Fos, caspase-3 and IL-1β expressions in the gastric antrum with that in the spinal cord in rats with cervical spondylosis, suggesting that the gastrointestinal function may be affected by cervical spondylosis.     

Expression of c-fos in gastric myenteric plexus and spinal cord of rats with cervical spondylosis

AIM: To determine the expression of c-fos in gastric myenteric plexus and spinal cord of rats with cervical spondylosis and its clinical significance. METHODS: A cervical spondylosis model was established in rats by destroying the stability of cervical posterior column, and the cord segments C4-6 and gastric antrum were collected 3, 4 and 5 mo after the operation. Rats with sham operation were used as controls, c-fos neuronal counter-staining was performed with an immunohistochemistry method. Every third sections from C4-6 segments were drawn. The 10 most labeled c-fos-immunoreactive (Fos-IR) neurons were counted, and the average number was used for statistical analysis. The mean of Fos-IR neurons in myenteric plexus was calculated after counting Fos-IR neurons in 25 ganglia from each antral preparation, and expressed as a mean count per myenteric ganglion. RESULTS: There were a few c-fos-positive neurons in the cervical cord and antrum in the control group. There was an increased c-fos expression in model group 3, 4 and 5 mo after operation, whereas there was no significant increase in c-fos expression in the control group at 3, 4 and 5 mo. More importantly, there was a significant difference in c-fos expression between rats followed up for 3 mo and those for 5 mo in the model group (11.20±2.26 vs 27.68±4.36, P<0.05, for the cervical cord; and 11.3±2,3 vs 29.3±4.6, P<0.05, for the gastric antrum). There was no significant difference between rats followed up for 3 mo and those for 4 mo and between rats followed up for 4 mo and those for 5 mo in the model group. CONCLUSION: c-fos expression in gastric myenteric plexus was dramatically associated with that in the spinal cord in rats with cervical spondylosis, suggesting that the gastrointestinal function may be affected by cervical spondylosis. If this hypothesis is confirmed by further studies, functional gastrointestinal diseases such as functional dyspepsia and irritable bowel syndrome could be explained by neurogastroenterology.     

毒素型及免疫型大鼠纤维化肝组织中金属蛋白酶组织抑制因子定位及表达差异

目的 探讨人血自蛋白免疫诱导型及四氯化碳(CCl4)毒素诱导型所致大鼠肝纤维化模型肝组织中金属蛋白酶组织抑制因子(TIMP)定位及表达状态的差异。方法 分别以20％人血白蛋白进行免疫攻击和CCl4毒素攻击大鼠,造模结束后对大鼠肝组织进行H-E、V-G染色及电镜观察;同时研究TIMP-1、TIMF-2蛋白及mRNA定位及表达状态。结果 造馍结束后,免疫诱导型模型肝组织病理改变具有进行性加重趋势,TIMP-1、TIMP-2 mRNA及蛋白表达强度高、持续时间长：而毒素诱导型模型具有TIMP-1、TIMP-2mRNA及蛋白表达强度弱、肝纤维化持续时间短等特点。免疫诱导实验组肝脏中TIMP-1、TIMF-2相关抗原表达在肌成纤维细胞、成纤维细胞,以汇管区及纤维间隔中最明显,阳性信号位于细胞胞质中,未见细胞核表达。原位杂交检测结果亦相似。结论 毒素诱导型肝纤维化模型自然吸收较快,不利于抗肝纤维化药物疗效的观察;免疫诱导型肝纤维化模型自然吸收时间较慢,且在造模结束后1～3个月有逐渐加重趋势,有利于抗肝纤维化药物疗效观察及肝纤维化机制研究。     

Investigations of fluorescent peptides and lipofuscins of human intervertebral disc relating to atherosclerosis.

(1) Annulus fibrosus and nucleus pulposus of human intervertebral discs at different degrees of atherosclerosis were disintegrated by elastase. (2) The material disintegrated by elastase -- called elastolysate -- could be separated into hydrophobic (apolar) and hydrophilic (polar) peptides. Parallel with the degree of atherosclerosis the amount of hydrophobic peptides increased, whereas that of the hydrophilic peptides decreased. (3) In annulus fibrosus and nucleus pulposus two kinds of fluorescence maxima were measured. The one, A:F 350:405, is known as fluorescence maxima of elastin- and collagen-peptides. The other, A:F 410:470, is related to a similar substance called atherofluorescent component (AFC), which has been isolated before from the plaques of atherosclerotic aorta. This substance accumulates mainly in nucleus pulposus and resembles lipofuscin-like bodies. (4) These bodies show a positive reaction with thiobarbituric acid, giving a red coloration characteristic for malondialdehyde. In nucleus pulposus the amount of lipofuscin-like substances is much greater than in annulus fibrosus. (5) The hydrophilic peptides, although they show the same fluorescence maxima as the hydrophobic peptides, do not give any reaction with thiobarbituric acid. It is supposed that in these cases the cross-linked protein contains instead of malondialdehyde other reactive aldehydes.     

耳压加经颅磁刺激治疗椎动脉型颈椎病疗效观察

目的探讨耳压加经颅磁刺激治疗椎动脉型颈椎病的临床疗效。方法将98例椎动脉型颈椎病患者随机分为2组,治疗组53例采用耳压加经颅磁刺激治疗,对照组45例采用药物治疗。结果治疗组的总有效率和好转率都高于对照组,2组相比有显著性差异（P〈0．05）．结论耳压加经颅磁刺激治疗椎动脉型颈椎病疗效显著,可以在临床推广。     

经额部入路注射自体动脉血法构建大鼠尾状核脑出血模型

目的探讨采用经额部入路2次注射自体动脉血的立体定向手术方法,改进大鼠尾状核脑出血模型的可行性。方法 SD大鼠16只,分为经额部入路组(8只)和经顶部入路组(8只)。2组分别在立体定向仪辅助下,自大鼠额部以及顶部入路进针至左侧尾状核中心,取股动脉血50μl分2次注入尾状核,观察比较2种不同方法所致血肿的形态及容量,并进行统计分析。结果所有大鼠左侧尾状核内均可见血肿形成。经额部入路组大鼠平均血肿容量(25.1±0.4)μl,明显多于经顶部入路组(24.2±1.0)μl,差异有统计学意义(P0.05)。2组血肿容量离散度分别为1.4%和4.2%。经额部入路组大鼠血肿形态较规则,优于经预部入路组。结论经额部入路2次注射自体动脉血技术是构建稳定大鼠尾状核脑出血模型的有效手段。     

胃食管反流病与颈椎病相关性研究

目的探讨颈椎病患者是否为胃食管反流病(gastroesophageal reflux disease,GERD)的高发人群。方法 321例经临床及磁共振诊断为颈椎病的患者作为颈椎病组,同期选取249名健康体检者作为对照组,进行RDQ评分问卷调查,比较颈椎病组及对照组的RDQ评分及GERD发生率差异是否有统计学意义,颈痛程度与RDQ评分是否相关,并对可能危险因素及颈椎病分型情况进行统计学分析。结果颈椎病组RDQ评分明显高于对照组(P0.05);以RDQ评分≥12分作为GERD评判标准,颈椎病组GERD 43例(13.4%),对照组GERD 17例(6.8%),颈椎病组GERD发生率明显高于对照组(P0.05);不同颈痛程度组间RDQ评分差异有统计学意义(P0.05);各型颈椎病中,交感神经型发生GERD的风险明显高于其他各型(P0.05);颈椎病合并GERD的危险因素主要为女性、非甾体抗炎药和精神心理因素。结论颈椎病患者发生GERD的风险较高,且以交感神经型最为显著。女性、药物和精神心理因素是颈椎病合并GERD的主要危险因素。     

经皮椎间孔镜在治疗中老年腰椎间盘突出症中的应用

目的评价应用经皮椎间孔镜(PTED)治疗中老年腰椎间盘突出症的临床效果和价值。方法前瞻性研究分析2016年3月至2018年9月空军特色医学中心骨科采用PTED切除突出椎间盘髓核减压治疗的50～72岁腰椎间盘突出症患者,共50例,并以同期常规开放椎板开窗或半椎板切除减压突出椎间盘髓核切除手术的50～71岁患者50例为对照组。比较2组患者术中出血量、手术时间、术后住院时间。以术后1 d、1个月、3个月疼痛视觉模拟评分(VAS)和术后3个月Oswestry功能障碍指数(ODI),及术后6个月改良MacNab标准评定手术疗效。手术前及术后3～6个月行MRI检查,观察手术前后椎管及椎间盘突出的变化。使用STATA 12.0统计软件进行统计分析。结果 2组患者年龄、术前VAS和ODI评分差异无统计学意义(P0.05),术后VAS和ODI评分较术前均显著下降(P0.01);PTED组术后1 d、1个月的VAS评分显著低于对照组(P0.01)。术后3个月,2组VAS和ODI评分差异无统计学意义(P0.05);术后6个月,2组改良MacNab标准评定功能差异无统计学意义(P0.05);PTED组术中出血量、术后住院时间显著少于对照组(P0.01);2组手术时间无统计学差异(P0.05)。MRI检查结果显示,与术前比较,2组术后3～6个月椎间盘突出均消失或明显减小,椎管通畅、面积明显增大。结论 PTED技术治疗中老年腰椎间盘突出症,中期疗效与单纯开放手术相当,但PTED早期疗效好、损伤小、恢复快、并发症少。     

复方黄芪颗粒抗肝纤维化机制的实验研究

目的:探讨复方黄芪颗粒(FFHQKL)有无抗肝纤维化作用及作用机制.方法:应用Chevallier半定量计分系统,病理组织学观察,结合细胞外基质(ECM)特殊染色动态观察高、低剂量FFHQKL治疗猪血清诱导肝纤维化模型后各时间段肝组织学及ECM量的变化,评估复方黄芪颗粒的抗肝纤维化疗效;应用免疫组织化学染色方法观察造模结束时以及药物治疗各时间段肝组织内基质金属蛋白酶-13(MMP-13)及其抑制剂(TIMP-1)表达量的变化.结果:与模型组比较,高、低剂量FFHQKL治疗2个月、3个月结束时,肝组织Chevallier纤维化评分均明显减少(P＜0.01).FFHQKL高、低剂量治疗组在治疗第2个月结束时,TIMP-1蛋白表达明显减弱,MMP-13蛋白表达明显增强,治疗第3个月结束时,上述改变更加显著,与模型组比较差异有显著性意义(P＜0.05或P＜0.01).结论:FFHQKL可在蛋白水平增强MMP-13酶蛋白的表达,同时抑制TIMP-1酶蛋白的表达,促进ECM的降解,从而达到逆转肝纤维化之疗效.     

电针灸配合中药湿热敷治疗颈椎病的疗效观察

目的探讨电针灸配合中药湿热敷治疗颈椎病的临床效果。方法将我院收治的颈椎病患者60例随机分成观察组和对照组,各30例,观察组给予电针灸配合中药湿热敷治疗,对照组仅单独给予中药湿热敷治疗,观察两组治疗结果。结果观察组总有效率为100.0%,对照组总有效率为43.3%,两组比较差异有统计学意义(P<0.05)。结论电针灸配合中药湿热敷治疗颈椎病效果显著,值得临床推广应用。