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

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

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。

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: ①设备简单、费用低。②检测方法简便易行、迅速可靠。③无创伤性。④重复性好。⑤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“toe clearance"。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，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 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 from the severe group.)
B - Random formation of experimental groups
C - Spinal Cord Injury induced by weight drop controlledby the NYU
Impactor
1 Anesthesia
2 Laminectomy
3 Spinal cord contusion
D - General standard procedures after spinal cord contusion
E - Postoperative antibiotic therapy
F - Maintenance of the animals
G - Locomotor evaluation (simultaneous filming of the motricity of each rat by three 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 evaluators blinded to the degree of severity of each rat's injury).
H - Euthanasia 29 days after the injury
I - Statistical analysis
BBB SCALE

O - 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 extensive 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.
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 with weight bearing and no plantar stepping.
10. 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-hindlimb 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 locomotion 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 coordination 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 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 gait and movement of the toes occurs frequently during forward movement of 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 predominant 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 is 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 movement 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 position is predominantly parallel to the body during the whole support stage; consistent trunk stability; consistent tail elevation.

STATISTICAL ANALYSIS

In 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 Pearson's correlation coefficient (less than 30%). Since no normal distributions were found, non-parametric tests were adopted. We also used Wilcoxon's test for non-parametric paired samples to infer the difference of the means between the right (R) and left (L) sides of each rat, and Spearman's unilateral correlation coefficient (r) to check for pairing effectiveness. We used the Friedman test for non-parametric paired samples to infer the difference of the means among the different evaluators. The paired differences were discriminated using the multiple comparison test modified by Dunn. In those cases where the Dunn's test did not show enough statistical power (power effectiveness) to discriminate the differences, we minimally reported the difference between those evaluators who presented the greatest difference in ranking.

A 5% confidence level (a= 0.05) was used, as well as two-tailed tests (H0 = m1 - m2 = O).