Basso-Beattie-Bresnahan(BBB)脊髓损伤的行为学评分

1、脊髓损伤的行为学评分1、 BBB 法 评估大鼠后肢运动功能的恢复情况。将动物放置于平台上, 观察记录其后肢的行走及肢体活动。评分分三部分第一部分为0 一 7 分 , 评判动物后肢各关节活动第二部分为8 一 13 分 , 评判后肢的步态及协调功能第三部分为14 一21 分 , 评判运动中爪的精细动作 , 三项满分为21 分 . 实验动物分别在损伤后第周进行评分Basso Beattie Bresnahan(BBB) 运动功能评分及脚印分析术前 3 d ，每天由 2 人分别对动物进行BBB 运动功能评分。 术后 1 d 、 3 d 及每周对动物进行BBB 评分。 评分 2 人不了解实验进程及分组情

2、况。 在实验过程中，对 BBB 评分大于8 分的动物进行脚印分析：将动物的前后足分别用绿红2 种染料标记后，置于预先铺有白纸的7.5 cm 100 cm 的跑道中，使动物从一端跑到另一端，计算大鼠同侧前后足中心的距离(interlimb coordination ， ILC) 及后肢第 3 足趾的外旋角度(angle of rotation ， AR)进行分析。B B B 表评分项目表分数特征说明0无可观察到的后肢运动HL：(hind-limb ， HL)后肢11或 2个 HL关节轻度活动轻度：5%关节活动范围21个 HL关节大幅运动和另一关节轻度活动大幅：50%关节活动范围32个 HL关节大

3、幅运动2个关节：通常为髋和膝4所有3个 HL关节轻度活动3个关节：髋、膝、踝；第3个关节通常为踝52个 HL关节轻度运动和第3个关节大幅运动62 个 HL 关节大幅运动, 第 3 个关节轻度活动7所有3个 HL关节大幅运动8不负重拖动或足置于不负重位拖动：节律性伸展3个 HL关节, 身体侧卧9足底仅位于负重位, 或偶尔/ 频繁/持续以足背负重步行, 无足底负重步行负重：足底负重位时或仅在后躯干抬高时,HL伸肌收缩10偶见负重步行, 但前、后肢不协调偶尔: 5%且 50%：步行：足底负重触地，HL前置使足底再次触地11由频繁到持续负重步行, 但无前一后肢协调频繁：51%-94%观察期；持续：95

4、%-100%观察期12由频繁到持续负重步行, 偶见前一后肢协调6%-50%协调运动13由频繁到持续负重步行, 频繁前一后肢协调51%-95%协调运动14持续协调足底步行, 优势爪在刚触地和抬起时旋转频繁足底步行, 持续前一前肢协调, 偶尔足背侧旋转：当其触地或抬起时后爪内或外旋15持续协调足底步态, 当前肢向前时无或偶有伸趾, 优势爪在刚触地时平行平行：后爪在刚触地或抬起时与躯干平行 伸趾：听趾，拖踏音，及足音中无趾拖踏音16持续协调足底步态, 频繁伸趾, 优势爪触地时平行，抬起时旋转频繁伸趾：一半以上足音中无趾拖踏音17持续协调足底步态, 频繁伸趾, 优势爪在触地及抬起时均平行18持续协调足

5、底步态, 持续伸趾, 优势爪在触地及抬起时均平行持续伸趾：4min观察期中仅有4次趾拖踏音19基本内容同18，尾在部分或全部观察期中垂20基本内容同18，尾持续上翘，但躯体不稳尾上翘：不触地：躯体不稳：当快速移动时, 重心侧移 , 出现摇摆、倾斜、滑倒21基本内容同20，且躯体持续稳定躯体持续稳定：无滑倒, 骨盆环与尾在运动时保持一直线2、 斜 板 实验 (Rivline ta l.,1 977):总体评估四肢肌力。斜板表面垫以6mm厚的橡胶垫，按大鼠身体轴线与斜板纵轴垂直的方向放置大鼠，逐渐增加斜板与水平面间的角度，直至大鼠刚好可在板上停留5s，记录这一角度。每只大鼠测3次， 取平均值，所有

6、行为学评估均在单盲情况下进. 评分时间分别为损伤后第周进行斜板实验的优点有: 设备简单、费用低。 检测方法简便易行、迅速可靠。无创伤性。重复性好。与脊髓损伤程度相关性较强。因而是目前较为常用的方法。其缺点是对运动功能评价较为单一, 存在人为因素BBB 评分表是根据观察脊髓损伤大鼠经过三个阶段的恢复而建立的。1、 早期：以无或极少的后肢关节运动为特征。2、 中期：包括几次共济失调步态。3、 晚期：包括精细运动，如拖着脚趾和尾巴，躯干不稳定以及爪子交替轮转。术后第 1 天进行的旷场试验。可以观察到后肢运动受限，动物无力支撑体重以至于拖着躯干、后腿和臀部。这是恢复阶段的早期。恢复阶段的中期在此呈现的

7、是术后第16 天，动物可以走路以及支撑自己的体重。在这个阶段，前后肢的协调运动恢复，大鼠能持续行走。恢复阶段的后期在此呈现的是术后第41 天。在这个阶段，一些精细运动恢复.种类的说明大体上， 21 种评分表与恢复的进展是同步的。第一部分，0-7类，评估恢复早期的后肢关节运动。0. 未见后肢运动1. 一个或两个关节的轻微运动，通常是髋关节和/或膝关节2. 一个关节的广泛运动或一个关节的广泛运动加上其它关节的轻微运动3. 两个关节的广泛运动4. 后肢三个关节的轻微运动（髋关节，膝关节和踝关节）5. 两个关节的轻微运动和另一个关节的广泛运动6. 两个关节的广泛运动和另一个关节的轻微运动7. 后肢三个

8、关节的广泛运动第二部分，8-13 类，评估恢复中期的步态和协调运动。8. 无负重拖动或足置于无负重位9. 足底仅位于负重位，或偶尔频繁持续的足背负重步行，无足底步行10. 偶尔负重步行，无前后肢协调运动11. 频繁到持续的负重步行，无前后肢协调运动12. 频繁到持续的负重步行，偶有前后肢协调运动13. 持续的负重步行，频繁的前后肢协调运动第三部分，14-21 类，评估运动时爪子的精细运动。14. 持续协调步态，持续前后肢运动协调；运动时优势爪旋转或频繁足底步行，持续前后肢运动协调和偶尔的足背步行。15. 持续协调步态；当前肢前进时无或偶有伸趾；优势爪刚触地时与身体平行。16. 持续协调步态；频

9、繁伸趾；优势爪触地时与身体平行，提起时旋转。17. 持续协调步态；频繁伸趾；优势爪在触地及提起时均与身体平行。18. 持续协调步态；持续伸趾；优势爪在触地与身体平行，提起时旋转。19. 持续协调步态；持续伸趾；优势爪在触地及提起时均与身体平行。20. 持续协调步态；持续伸趾；优势爪在触地及提起时均与身体平行；但躯体不稳定，尾巴持续上翘。21. 持续协调步态；持续伸趾；优势爪在触地及提起时均与身体平行；躯体稳定，尾巴持续上翘。其中特别注意的有一个UNIQUE MOVEMENT ，最费解的是伸趾，即“ toeclearance。在评分时，这项运动是用听的，而不是看的，如果听到有爪子在地面刮的声音，

10、越频繁则表示toe clearance越少（即两者的频率表示是相反的）。所以，以前的中文BBB 评分中把 toe clearance直接翻译成“爪抓地 ”是不恰当的。Quantification of BBB Scores. The BBB 21-point open-field locomotor rating scale was used for evaluating hindlimb movement (15, 3). In our experiments, the rats were evaluated every day for the first 3 days and every

11、3.5 days for the remaining 6 weeks after injury.A - Receipt and selection of animals (since a high number of rats died before 28 days after spinal cord injury, itwas necessary to begin work with 60 rats. Ultimately, eight rats were excluded from the mild group, nine from the moderate group and 13 fr

12、om the severe group.)B - Random formation of experimental groupsC - Spinal Cord Injury induced by weight drop controlledby the NYUImpactor1 Anesthesia2 Laminectomy3 Spinal cord contusionD - General standard procedures after spinal cord contusionE - Postoperative antibiotic therapyF - Maintenance of

13、the animalsG - Locomotor evaluation (simultaneous filming of the motricity of each rat bythree digital cameras 28 days after mild, moderate or severe spinal cord injury;video-based analysis and corresponding BBB scale assessment of the locomotor functional capacity was conducted by six independent e

14、valuators blinded to the degree of severity of each rats injury).H - Euthanasia 29 days after the injuryI - Statistical analysisBBB SCALEO - No observable movement of the hindlimbs.1. Slight (limited) movement of one or two joints, usually hip and/or knee.2. Extensive movement of one joint or extens

15、ive movement of one joint and slight movement of the other.3. Extensive movement of two joints.4. Slight movement of all three joints of the hindlimbs.5. Slight movement of two joints and extensive movement of the third joint.6. Extensive movement of two joints and slight movement of the third joint

16、.7. Extensive movement of the three joints in the hindlimbs.8. Sweeping without weight bearing or plantar support of the paw without weight bearing.9. Plantar support of the paw with weight bearing only in the support stage (i.e., when static) or occasional, frequent or inconsistent dorsal stepping

17、with weight10. Plantar stepping with occasional weight bearing and no forelimb-hindlimb coordination.11. Plantar stepping with frequent to consistent weight bearing and occasional forelimb-hindlimb coordination.12. Plantar stepping with frequent to consistent weight bearing and occasional forelimb-h

18、indlimb coordination.13. Plantar stepping with frequent to consistent weight bearing and frequent forelimb-hindlimb coordination.14. Plantar stepping with consistent weight support, consistent forelimb-hindlimb coordination and predominantly rotated paw position (internally or externally) during loc

19、omotion both at the instant of initial contact with the surface as well as before moving the toes at the end of the support stage or frequent plantar stepping, consistent forelimb-hindlimb coordination and occasional dorsal stepping.15. Consistent plantar stepping, consistent forelimbhindlimb coordi

20、nation and no movement of the toes or occasional movement during forward movement of limb; predominant paw position is parallel to the body at the time of initial contact.16. Consistent plantar stepping and forelimb-hindlimb coordination during gait and movement of the toes occurs frequently during

21、forward movement of the limb; the predominant paw position is parallel to the body at the time of initial contact and curved at the instant of movement.17. Consistent plantar stepping and forelimb-hindlimb coordination during gaitand movement of the toes occurs frequently during forward movement of

22、limb; the predominant paw position is parallel to the body at the time of initial contact and at the instant of movement of the toes.18. Consistent plantar stepping and forelimb-hindlimb coordination during gait and movement of the toes occurs consistently during forward movement of limb; the predom

23、inant paw position is parallel to the body at the time of initial contact and curved during movement of the toes.19. Consistent plantar stepping and forelimb-hindlimb coordination during gait and movement of the toes occurs consistently during forward movement of limb; the predominant paw position i

24、s parallel to the body at the instant of contact and at the time of movement of the toes, and the animal presents a downward tail some or all of the time.20. Consistent plantar stepping and forelimb-hindlimb coordination during gait and movement of the toes occurs consistently during forward movemen

25、t of limb; the predominant paw position is parallel to the body at the instant of contact and at the time of movement of toes, and the animal presents consistent elevation of the tail and trunk instability.21. Consistent plantar stepping and coordinated gait, consistent movement of the toes; paw pos

26、ition is predominantly parallel to the body during the whole support stage; consistent trunk stability; consistent tail elevation.STATISTICAL ANALYSISIn order to analyze the sensitivity and reproducibility of the results, we checked for consistency between the results achieved on the right and left sides of the rats and between the grouped results achieved by the different evaluators.The normality of the distributions was tested using the Kolmogorov-Smirnov test for continuous variables and by examining the Pearsons correlation coeffici