NGF对哮喘豚鼠下呼吸道及内脏感觉传入部位IL-1β表达的影响

为探讨神经生长因子(NGF)在哮喘发病中的作用及其神经免疫调节机制,本研究应用免疫组织化学方法检测各组豚鼠下呼吸道和内脏感觉传入部位白介素-1β(IL-1β)免疫反应的变化。用Western blot方法分别检测各组豚鼠肺、C7~T5段脊神经节和相应节段脊髓背角NGF和IL-1β的表达。用Luzex-F实时图像分析系统和凝胶成像分析系统分别对结果进行图像分析。免疫组织化学结果显示(染色反应强度用灰度值表示):生理盐水组与单纯致敏组豚鼠IL-1β阳性免疫反应产物的平均灰度值没有显著差异(P>0.05);与这两个对照组相比,哮喘豚鼠气道上皮、肺内炎性细胞、血管平滑肌内的神经末梢、C7~T5段脊神经节及相应节段脊髓背角内IL-1β阳性反应产物的平均灰度值均明显降低(P<0.01);在哮喘+NGF抗体组(鼻腔吸入NGF抗体),上述部位内IL-1β阳性反应产物的平均灰度值均明显高于哮喘组(P<0.01)。Western blot结果显示:与生理盐水组和单纯致敏组比较,哮喘组豚鼠肺、C7~T5段脊神经节及相应节段脊髓样品中IL-1β或NGF目标带的IDV(integrated density value)与内参照IDV的比值均明显升高(P<0.01),而哮喘+NGF抗体组上述部位样品中IL-1β或NGF目标带的IDV与内参照IDV的比值均明显低于哮喘组(P<0.01),表明NGF可上调IL-1β的表达。这些结果提示NGF诱发IL-1β的表达可能是NGF参与哮喘发病的机制之一,NGF抗体抑制哮喘时下呼吸道和内脏感觉传入部位IL-1β的表达可能是治疗哮喘的新途径。     

神经生长因子对哮喘豚鼠内脏感觉传入部位MMP-2表达的影响

目的探讨基质金属蛋白酶-2(MMP-2)在哮喘豚鼠内脏感觉传入部位的表达及神经生长因子(NGF)对MMP-2的调节作用。方法应用免疫组织化学方法检测豚鼠内脏感觉传入部位MMP-2免疫反应的变化。用W estern b lot蛋白印迹方法检测豚鼠C7~T5段脊神经节和相应节段脊髓背角NGF和MMP-2的表达。用Luzex-F实时图像分析系统和凝胶成像分析系统分别对结果进行图像分析。结果①免疫组织化学结果显示:豚鼠C7~T5段脊神经节和相应节段脊髓背角内MMP-2阳性免疫反应产物的平均灰度值(0-深色,255-浅色),生理盐水组(175.50±4.67、166.47±6.20)与单纯致敏组(172.63±9.51、165.20±5.14)比较没有显著差异(P>0.05);哮喘组(139.87±4.88、125.93±4.49)明显低于生理盐水组和单纯致敏组(P<0.01);哮喘+NGF抗体组(鼻腔吸入NGF抗体,169.73±6.24、152.37±8.67)则明显高于哮喘组(P<0.01)。②W estern b lot结果显示:与生理盐水组和单纯致敏组比较,哮喘组豚鼠C7~T5段脊神经节及相应节段脊髓样品中MMP-2和NGF阳性产物的IDV(Integrated Density Value)与β-actin IDV的比值均明显升高(P<0.01),而哮喘+NGF抗体组则明显低于哮喘组(P<0.01),表明NGF可上调豚鼠内脏感觉传入部位MMP-2的表达。结论C7~T5段脊神经节以及相应节段脊髓背角内的MMP-2可能也参与哮喘的发病过程,NGF诱发MMP-2的表达可能是NGF参与哮喘发病的机制之一,NGF抗体抑制哮喘时MMP-2的表达有可能延缓气道重塑的发生,可能是治疗哮喘的新途径。     

哮喘豚鼠下呼吸道及内脏传入部位NGF表达的研究

目的　探讨哮喘豚鼠下呼吸道和内脏传入系统神经生长因子 (NGF)的作用。　方法　利用免疫组织化学和原位杂交结合显微图像分析 ,研究哮喘豚鼠下呼吸道和内脏传入系统NGF免疫反应及mRNA表达的变化。　结果　与对照组相比 ,哮喘豚鼠NGF免疫反应在气道上皮、C7～T5段脊神经节及脊髓背角明显上调 (P <0 0 1) ,NGFmRNA表达在气道上皮明显增多 (P <0 0 1) ,而在C7～T5段脊神经节及脊髓背角却未见明显改变。　结论　气道上皮、C7～T5段脊神经节及对应节段脊髓背角的NGF可能参与哮喘的发病过程 ;C7～T5段脊神经节细胞及对应节段脊髓背角神经细胞本身能合成NGF。     

地塞米松对哮喘豚鼠肺内、脊神经节及脊髓背角内蛋白激酶C表达的抑制作用

目的探讨地塞米松对哮喘豚鼠肺内、C7～T5脊神经节及对应节段脊髓背角内蛋白激酶C（PKC）表达的抑制作用。方法利用免疫组织化学和Western blotting方法。研究哮喘豚鼠肺内、C7～T5脊神经节及对应节段脊髓背角内PKC的免疫反应变化。结果哮喘组豚鼠肺内、C—L脊神经节及对应节段脊髓背角内PKC平均光密度值明显高于对照组（P〈0．01）,而地塞米松组则明显低于哮喘组（P〈0．01）。结论PKC可能参与哮喘的发病过程,地塞米松抑制哮喘豚鼠PKC的表达可能是其治疗哮喘的作用机制之一。     

Akt对哮喘小鼠下呼吸道及内脏感觉传入部位IL-1β表达的影响

目的探讨在哮喘发病机制中,PKB/Akt对哮喘小鼠肺组织及C7-T5段脊神经节和相应节段脊髓后角IL-1β表达的调节作用。方法BALB/c小鼠30只,按随机数字表法均分为正常对照组、哮喘组、PKB/Akt阻断组,免疫组织化学检测各组小鼠肺组织及C7-T5节段脊神经节和相应节段脊髓后角IL-1β的表达,Western blot方法检测各组小鼠肺组织及C7-T5节段脊神经IL-1β的表达。结果免疫组织化学结果显示,哮喘组肺、C7-T5段脊神经节和相应节段脊髓后角IL-1β阳性产物的平均光密度值(MOD)显著高于正常对照组(P<0.01);而PKB/Akt阻断组与哮喘组相比MOD值明显降低(P<0.05)。Western blot结果显示:与正常对照组比较,哮喘组小鼠肺、C7～T5段脊神经中IL-1β的IDV(integrated density value)与β-actin IDV的比值均明显升高(P<0.01),而PKB/Akt阻断组明显低于哮喘组(P<0.05)。结论NGF介导的Akt通路的激活可上调IL-1β的表达,介导哮喘气道高反应和炎症反应。     

实验性哮喘时神经生长因子对P物质的上调作用

目的：探讨实验性哮喘时神经生长因子（NGF）对P物质（SP）的调节机制。 方法： 应用放射免疫分析方法检测了NGF缺失时（鼻腔吸入NGF抗体）哮喘豚鼠下呼吸道和内脏感觉传入部位SP含量的变化。 结果： NGF缺失时哮喘豚鼠气管、支气管、肺、C7-T5段脊神经节及相应节段脊髓后角、结状神经节和孤束核区SP含量明显低于哮喘组和对照组（P<0.01）。 结论： 实验性哮喘时NGF可上调下呼吸道和感觉神经节等内脏感觉传入部位的P物质水平，两者共同参与支气管哮喘的发病过程。     

NGF对哮喘豚鼠下呼吸道及内脏感觉传入部位TNF-αmRNA表达的上调作用

目的　探讨神经生长因子 (NGF)在哮喘发病中的作用及其神经免疫调节机制。　方法　应用原位杂交方法结合显微图像分析 ,研究实验性哮喘豚鼠下呼吸道和内脏感觉传入部位肿瘤坏死因子 αmRNA(TNF αmRNA)表达的变化以及NGF抗体对其的影响。　结果　生理盐水组与单纯致敏组TNF αmRNA的表达没有显著差异 (P >0 0 5 ) ;与这两个对照组相比 ,哮喘豚鼠TNF αmRNA的表达在气道上皮、肺内炎性细胞、C7～T5段脊神经节及脊髓后角内均明显上调 (P <0 0 1) ;在哮喘 +NGF抗体组 ,上述部位内TNF αmRNA的表达明显低于哮喘组 (P <0 0 1)。　结论　NGF诱发TNF α的表达可能是NGF参与哮喘发病的机制之一 ,NGF抗体抑制哮喘时下呼吸道和内脏感觉传入部位TNF αmRNA的表达可能是治疗哮喘的新途径。     

实验性哮喘时神经生长因子对神经激肽A的上调作用

目的:探讨实验性哮喘时神经生长因子(NGF)对神经激肽A(NKA)的调节机制。方法: 应用放射免疫分析方法检测了NGF缺失时(鼻腔吸入NGF抗体)哮喘豚鼠下呼吸道和内脏感觉传入部位NKA含量的变化。结果:NGF缺失时哮喘豚鼠气管、支气管、肺C₇-T₅段脊神经节及相应节段脊髓后角、结状神经节和孤束核区NKA含量明显低于哮喘组和对照组(P＜0.01)。 结论: 实验性哮喘时NGF可上调下呼吸道和感觉神经节等内脏感觉传入部位的神经激肽A水平,二者可能参与支气管哮喘的发病过程。     

地塞米松对哮喘豚鼠肺及内脏感觉传入系统神经生长因子表达的抑制作用

目的：探讨地塞米松对哮喘豚鼠肺及内脏感觉传人系统(C7-T5脊神经节及对应的脊髓后角)神经生长因子(NGF)表达的影响。方法：利用免疫组织化学和Westem印迹方法,观察生理盐水组、单纯致敏组、哮喘组和地塞米松组豚鼠肺及内脏感觉传人系统(C7-T5脊神经节及对应的脊髓后角)NGF表达的变化。结果：哮喘组豚鼠NGF在肺及内脏感觉传人系统(C7-T5脊神经节及对应的脊髓后角)表达明显高于对照组,而地塞米松组则明显低于哮喘组。结论：NGF可能参与哮喘的发病过程,地塞米松抑制哮喘豚鼠NGF的表达可能是其治疗哮喘的机制之一。     

地塞米松对哮喘豚鼠肺内脏感觉传入部位TrkA表达的抑制作用

为了探讨地塞米松对哮喘豚鼠肺内脏感觉传入系统(C7T5脊神经节及对应的脊髓后角)TrkA表达的影响,我们利用免疫组织化学和Westernblot方法,研究了哮喘豚鼠C7T5脊神经节及对应的脊髓后角TrkA的免疫反应变化。结果显示:哮喘组豚鼠C7T5脊神经节及对应的脊髓后角TrkA表达明显高于对照组(P<0.01),而地塞米松组则明显低于哮喘组(P<0.01)。结果提示TrkA可能参与哮喘的发病过程,地塞米松抑制哮喘豚鼠TrkA的表达可能是其治疗哮喘的作用机制之一。     

腰骶部脊神经根的临床解剖学

通过临床选择性脊神经后根切断术中观察与测量,为临床提供了重要的参考数据。腰骶神经前、后根解剖会合点至椎间孔距离平均为1.5～2.0cm。前后两根相比,后根明显较前根粗,其中以L_5后根最粗、L_2～S_1各后根内小束的数目为4～18束,平均为7束,临床分离后根时可分为7束,以便进行电刺激选择。这些资料为腰骶部选择性脊神经后根切断术和马尾神经的手术提供了重要依据。     

脊髓下段后索动脉供应的应用解剖

目的 为脊髓下段后索切开行周围神经游离移植或转位桥接术提供解剖学资料。方法 观测15具(30侧)成人尸体标本T_(10)-L_1脊柱平面的脊髓后动脉及动脉吻合的位置、管径及出现率。结果 脊髓后动脉在脊髓后外侧沟与后正中沟之间,呈连续或断续分布,出现率100％,平均管径为0.46mm,距后正中沟距离平均2.2mm,距后外侧沟距离平均为0.54mm。两侧间无显著差异。脊髓后动脉间吻合动脉的出现率为33％,平均管径0.40mm。结论 脊髓后动脉是脊髓下段后索区域的主要供血来源。脊髓后索纵行切口选择在离后正中沟1.5mm处损伤血管的可能性较小。     

Location of the spinal nucleus of the accessory nerve in the human spinal cord

The segmental extent and topography of the spinal nucleus of the accessory nerve (SNAN) was investigated in the adult human spinal cord. Transverse sections of segments between the lower medulla and C6 were stained with cresyl violet and the motor cell columns identified according to the numerical locations defined by Elliott (1942). The segmental extent and topography of the cervical part of column 2 resembled that previously described for the SNAN of primates.     

A variation of the tissue print technique for studying isolated spinal cord cells in situ

We present a novel variation of the tissue print technique which yields isolated spinal cells that retain their original topographical organization in situ. A light papain digestion combined with a low-melting-point agarose as the adhesive substrate is used to ‘print' a monolayer of cells from spinal cord slices. Printed cells are viable for studies using electrophysiological approaches. This technique provides a means to bridge the gap between properties of isolated cells and their topographical identity in spinal cord.     

Morphometric comparison of computed tomography to magnetic resonance imaging in the evaluation of the lumbar intervertebral foramina

The purpose of this study was to evaluate and compare the ability of computerized tomography (CT) and magnetic resonance imaging (MRI) as techniques to determine linear morphometric measurements of several parameters of the right and left lumbar intervertebral foramina (IVFs). Specifically, the greatest superior-to-inferior diameter and anterior-to-posterior diameter of the lumbar IVFs were measured in a cadaveric imaging phantom. The phantom was first measured directly with vernier calipers and then embedded in gelatin to simulate soft tissue. It was then scanned with two types of protocols each for CT and MRI. The scanned images were transferred directly from the imaging units to optical disks, which were then read using an optical disk drive. The measurements taken directly from the phantom were then taken from the scans using a Macintosh II computer interfaced with the optical disk drive. The results showed that both sagittally reformatted CT images and sagittal MRI images were clinically and statistically reliable and valid methods for linear evaluation of the IVF in the sagittal plane. However, measurements made from the MRI scans were found to be more accurate than those made from the CT scans. The results of this study should help increase understanding of the strengths and weaknesses of both imaging modalities in the sagittal evaluation of the lumbar IVFs. The results may also help with the future evaluation of the IVF in the condition known as nerve root canal stenosis. This study also indicates that a normal morphometric database from the MRI scans of normal human subjects should be developed. © 1994 Wiley-Liss, Inc.     

Changes in Spinal Canal Diameter and Vertebral Body Height with Age

Purpose

All structures of the spine, including the spinal canal, change continuously with age. The purpose of this study was to determine how the spinal canal of the lumbar spine changes with age. The L4/5 is the most common site of spinal stenosis and has the largest flexion-extension motion, whereas the T5/6 has the least motion. Therefore, we measured the spinal canal diameter and vertebral body height at T5, T6, L4, and L5 with age.

Materials and Methods

This was a retrospective study of aged 40 to 77 years. We reviewed whole spine sagittal MRIs of 370 patients with lumbar spinal stenosis (LSS) (Group 2) and 166 herniated cervical disc (HCD) (Group 1). Each group was divided into four age groups, and demographic parameters (age, gender, height, weight, BMI), the mid-spinal canal diameter, and mid-vertebrae height at T5, T6, L4, L5 were compared. Within- and between-group comparisons were made to evaluate changes by age and correlations were carried out to evaluate the relationships between all parameters.

Results

Height, weight, and all radiologic parameters were significantly lower in Group 2 than Group 1. Group 1 did not show any differences, when based on age, but in Group 2, height, weight, and T6, L4, and L5 height were significantly decreased in patients in their 70''s than patients in their 40''s, except for spinal canal diameter. Age was associated with all parameters except spinal canal diameter.

Conclusion

Vertebral height decreased with age, but spinal canal diameter did not change in patients with either LSS or HCD. Mid-spinal canal diameter was not affected by aging.     

胸腰脊神经后根形态计量研究

在15具成人尸体上对胸腰脊神经后根进行了大体解剖和形态计量研究.结果表明:(1)上胸段脊神经后根的囊外段长度、硬膜点横径和脊髓点束数在逐节减少;后根的囊外段与囊长轴之下夹角>90°,囊内段及脊髓点分布长度在逐节增加,交通支最丰富.(2)中胸段脊神经后根的囊外段长度、硬膜点横径和脊髓点束数各节段波动范围较小,下夹角在90°左右,囊内段短,脊髓点分布长.(3)下胸段脊神经后根的脊髓点分布长度转而下降,下夹角<90°,其它指标均逐节增加.(4)腰段脊神经后根的脊髓点分布长度进一步减少,下夹角最小,其它指标达最大值,交通支丰富.根据研究结果进行后根受损危险排序,腰>下胸>上胸>中胸.     

脊髓动脉的显微解剖

在手术显微镜下观察了37例婴幼儿脊髓前(正中)动脉和脊髓后(外侧)动脉的分支,其主要结果如下:婴幼儿脊髓平均长度为14.08±1.49cm,脊髓前(正中)动脉外侧分支平均为76.17±18.64支,中央动脉平均为169.78±25.51支.中央动脉分布到灰质前角、侧角和后角底部及邻近灰质的深层白质,而白质浅层和后角大部,则由软膜动脉网发出的穿支供应.此外.还讨论了与临床应用有关的问题.     

荧光显微镜观察封闭式脊髓窗内的大鼠脊髓微循环

目的 探讨应用荧光显微镜通过改进的封闭式脊髓窗,观察大鼠脊髓背侧软脊膜微循环状态的可行性及效果.方法 用8周龄雄性SD大鼠,暴露T9至T11棘突,咬除T10棘突和椎板,暴露硬脊膜；在T9和T11棘突上安装固定器辅助物.用牙科丙烯酸树脂和玻璃片为原料在T10节段安装改进的封闭脊髓窗.采血并离心(2 000 ×g)收集红细胞,将100 μL压积的红细胞用Dil荧光染料(5 mg/L)体外孵育,静脉注射人体内；用罗丹明6G(0.3 g/L)静脉注射,体内标记白细胞.大鼠脊柱用微循环观测椎体固定器进行固定.用动物活体荧光成像显微系统进行观测和记录.结果 通过该系统可以观测大鼠软脊膜上微血管的形态和分布特点、血管密度；可以观测到Dil标记的红细胞在血管内的黏附和流速,观察和记录罗丹明6G标记的白细胞在血管壁的黏附滚动.结论 改进式的封闭脊髓窗和微循环观测椎体固定器,可用于对大鼠软脊膜上微血管进行观察和检测.     

经椎弓根椎体截骨术治疗脊柱后凸畸形的临床疗效观察

目的：探讨经椎弓根椎体截骨脊柱缩短术治疗并发脊髓神经功能障碍的脊柱后凸畸形的临床疗效。方法将2013年5月至2014年6月期间我院收治的80例并发脊髓神经功能障碍的脊柱后凸畸形患者纳入研究对象,采用随机数字表法分为观察组和对照组各40例,观察组患者接受经椎弓根椎体截骨术治疗,对照组患者接受经椎板和小关节突截骨术,比较2组患者手术情况、椎体愈合和神经功能恢复、治疗效果。结果观察组患者手术时间[(76.52±9.1) min]、术后下床活动时间[(3.28±0.43) d]短于对照组[(113.46±13.44) min,(5.67±0.68) d],术中出血量[(36.14±4.28) mL]、术后引流量[(17.92±2.12) mL]少于对照组[(55.23±7.15) mL,(29.64±4.28) mL]。观察组患者的Cobb角、出现初始尿意和强烈尿意时的膀胱容量[(6.12±0.68) mL,(456.56±51.78) mL]、残余尿量[(241.45±28.56) mL,(63.78±7.24) mL]低于对照组[(9.78±1.21) mL,(335.54±36.86) mL,(586.35±63.12) mL,(96.32±10.22) mL],椎间隙高度[(12.62±2.81) mL]高于对照组[(8.41±1.32) mm]。观察组治疗优良率(97.50%)明显高于对照组(82.50%)。结论经椎弓根椎体截骨脊柱缩短术治疗有助于减小手术创伤,促进术后恢复,纠正后凸畸形,改善神经功能,提高治疗效果。