| Background: But at present, lack for blood in the brain estimating that medicine resists when injuring an effect , come reflect pesticide effect the change often adopt the brain to lack for blood queen series of morphology index , biochemistry index, less show solicitude for if the limb motion dysfunction that medicine arouses to lacking for the brain apoplexy unyielding and principled have improving an effect, this is that the brain lacks for blood curing the problem being hit by regard clinically exactly. Be necessary therefore building a set of behavior testing system, the ability estimating that the experiment lacks for motion of brain apoplexy unyielding and principled queen animal limb, reflects nerve blood vessel renovation degree thereby.Objective: The meticulous functional fore limb motion feasibility estimating that the bureau cooking stove nature brain lacks for the blood rat being put into much trouble reaches the explore the investigation and discussion pallet school assignment box and overshot testing box validity. Attempt to build behavior testing system, the cooking stove nature brain estimating that Chinese catalpa alcohol plays a game lacks for blood rat nerve behavior effect. The effect expressing , analytical catalpa observing catalpa diversity giving medicament amounts to generate 1 (Ang-1) , the blood vessel generate the plain acceptor (Tie-2) plain egg white to every group rat blood vessel urge the bureau cooking stove nature brain to lack for mature mechanism of blood rat freshman blood vessel.Methods:1. Tray task box preparation and standards of gradingTray task box preparation method: Consult document method; improve according to actual condition, preparation pallet school assignment box. Test a box being 28cm once grow 40cm, width, 26 high cm irons envelops. The iron distance between enveloping the positive vertical stroke iron balustrade is that 0.9cm, left have small Iron Gate, entrance providing a rat, the back uses blue color paper to seal off the back, colors forms contrast, the easy to have a picture taken precise writer with the rat. Iron steamer main front balustrade bottom installation has one wide 3 cm, 1 deep cm pallets, and use pure iron sheet to be made from. Will fill a city to full within the pallet selling mini melon seed when every time training or testing, the rat can stretch a fore limb grabbing melon seed through balustrade gap liberty. Tray task box tests standards of grading: Wait for a rat according to the document grabbing the mature stable action queen, train time contraction to be 10 min, the testing time just sets a limit for in 10 min inner hereafter. Break up overshot melon of rat seed process for 6 continuous action, be: (1) above stretching a fore limb till food; (2) toes open the inverted wrist overshot; (3) grips food; The elbow reclaims (4) limbs; (5) lying of fore limb and mouth coordinate a husking; (6) pair of the hind legs queen crouches, two fore limbs hold food mastication in both hands. Accomplish 6 continuous action for one time merely writing down a "success grabbing a (hit)"; Not grasping to food or food back fall but , be six above-mentioned links if the fore limb had stretched the balustrade gap, any link fails , write down then for "stretching claw (reach)". During training, finally, three time of average value "stretching claw"number of times ", grabbing " number of times successfully is each "the overshot rat success rate before making a model base line value ". According to the following, formula secretly schemes against: Success rate= ("grab number of times successfully"/ "stretching claw"number of times) *100.In the step-by-step preparation by screening training middle, bringing into will reach the following standard makes model range: (1) habituation uses the left fore limb, is "a left benefit hand"rat; (2) 10 overshot min success rate is equal to 65% greater than or about.Behind making model first 1d and making a model, 2d, 4d, 7d, 15d, 21d carry out that method appraising2. Grasping test box preparation and standards of gradingGrasping test box preparation method: Consult document method , "grab the testing box " according to actual condition improvement , preparation , test a box using the pressure favoring 5 mms to be made from gram strenuously paper of. The main body includes a tripartite: The passage main body, drawer style eat the slot and the anti-skidding slope. 25 long passage cm, width 6 cm, 5 high cm, allow the rat nothing to hinder from passing , whose turn-back is relatively difficult but. The passage top has the spiracle , both ends to have shrinking bolting the door , the rat easy to is unlike the direction entrance , opens up , closes to inclining shrinking once bolting the door on another one side when testing. In passage bottom board rectangle surface, that long in two edge and long near two edge of equal rank get along has the width 1cm gap respectively, makes the passage bottom board that the rat passes only broad 4cm. Owing a gap following is to eat a slot, eat slot depths for 3cm, strong point are 15.5 cm, one end has sealing off a warehouse, force the rat entrance passage inner to grab mini melon seed. Eat the slot top have cunning cover, the gap opening up time and passage fits , form entire passage with passage when being closed down. Eat a slot to command on dyadic drawer implantation passage down part, in an integral whole above passage being in. In passage, both ends is shrink bolt the door place having ready-made 30 with the floor; The anti-skidding slope , the rat easy to crawl entering passage , favoring may observe the rat fore limb at the same time acts.Grasping test box testing standards of grading: Wait for a rat according to the document grabbing the mature stable action queen, training time shortens for 10 min, the testing time just sets a limit to in 10 hereafter the min inner. Break up overshot melon of rat seed process for 6 continuous action, be: (1) above stretching a fore limb till food; (2) toes open the inverted wrist overshot; (3) grips food; The elbow reclaims (4) limbs; (5) lying of fore limb and mouth coordinate a husking; (6) pair of the hind legs queen crouches, two fore limbs hold food mastication in both hands. Accomplish 6 continuous actions for one time merely writing down a "success grabbing a (hit)"; but do not grasp to food or food back fall if the fore limb had stretched the passage bottom board gap, any link failure, writes down six namely above-mentioned links then for "stretching a claw (reach)". During training, finally, three time of average value "stretching claw"number of times ", grabbing"number of times successfully is each "the overshot rat success rate before making a model base line value ". According to the following, formula secretly schemes against: Success rate = ("grab number of times successfully"/ "stretching claw"number of times) *100.In the step-by-step preparation by screening training middle, bringing into will reach the following standard makes model range: (1) habituation uses the left fore limb, is "a left benefit hand"rat; (2) 10 overshot min success rate is equal to 65% greater than or about.Behind making model first 1d and making a model, 2d, 4d, 7d, 15d, 21d carry out that method appraising3. Balance beam fabrication and standards of gradingThe balance beam fabrication: Consult document method; improve according to actual condition, preparation balance beam tests a mould. The mould uses wood to be made from, divides three parts: Balance beam, holder, platform. Balance beam is 2.5cm grow 80cm, width, and method of 2.5 high cm wood sticks, go on foot smoothing face to face. 1/3 is got along besides two holders are located in balance beam , balance beam is risen to distance floor 1m place , is fixed on the down part, 10cm foams make bedding with height in the floor , prevent a rat from falling and hurting self from dropping on balance beam. Platform is 25 long cm, for a short time square width 25cm boxes, the initiation place and destination being balance beam get along.Balance beam tests standards of grading: Consult a document , revising , working out the standard grading as follows slightly: Be 0 mark: Pass through balance beam , can not fall over; 1 divide: Pass through balance beam, and get the smooth phenomenon of foot appears in the distance travelled being smaller than 50%'s (to be to fall over chance less than 50%); 2 divide: Pass through balance beam, only have greater than to smooth phenomenon of foot appear on 50%'s distance travelled middle (being chance greater than fall over 50%); 3 divide: Can pass through balance beam, but paralysis oblique tones hind legs being able to not help to move forward; 4 divide: Can not pass through balance beam, only may sit in balance beam above; 5 divide: Readjusting oneself to a certain extent with the rat on balance beam may fall down.Screens gradually in the training, will achieve the following standard to integrate makes the mold scope: (1) each time can smooth through the balance beam, does not have the big mouse which on the balance beam routine pauses; (2) walks the posture meet standards, namely: The back slightly toward the camber, the tail goes up.This method in after making the mold 2d, 4d, 7d, 15d, 21d carries on the evaluation4. The animal divided into group, making a model and measuring judge method feasibility examinationThe experiment animal is divided into group: Take 15 rats the random divides 5 set, be regular group , fake operation group , model group , normal saline group , compatriot two phosphorus's group, method is eaten according to document standard.The bureau cooking stove brain lacks for blood rat model preparation: Up to the standard big mice bring into making model range. Adopt right nice oblique tones low-order to open a skull, the electricity curdles to olfactory tract on the right side and a segment of cerebrum between the veins is hit by an arterial under the cerebrum, the preparation permanency bureau cooking stove sex brain lacks for the blood model.The tray task box measures a pallet judging method learning examination: Mg/2ml choosing specification as 250, dosages use normal saline to compound for 0.4g.kg-1 compatriot two phosphorus's gallbladders alkali injects, accumulate for 3 ml for medicine population every time, the normal saline gives an equivalent volume set , the abdominal cavity gets a shot. The model group and the fake operation group rat do not give medicine. 24h begin to medicine time from making a model, continue giving medicine 7 day. The 2nd , 4 , 7 , 14 and 21 d, uses pallet school assignment box to appraise every group rat left fore limb (be a limb be put into much trouble) grabbing ability of melon seed. Grab action, import a computer with telecamera itinerary rat, success rate analyzing and calculating every rat grabbing.The box measures the overshot testing judging method learning examination: Mg/2ml choosing specification as 250, dosages use normal saline to compound for 0.4g.kg-1 compatriot two phosphorus's gallbladders alkali injects, accumulate for 3 ml for medicine population every time, the normal saline gives an equivalent volume set , the abdominal cavity gets a shot. The model group and the fake operation group rat do not give medicine. 24h begin to medicine time from making a model, continue giving medicine 7 day. The 2nd , 4 , 7 , 14 and 21 d, with grabbing the ability testing a box appraising every group rat left fore limb (be a limb be put into much trouble) grabbing melon seed. Grab action, import a computer with telecamera itinerary rat, success rate analyzing and calculating every rat grabbing.5. Catalpa pesticide effect estimatesThe experiment animal is divided into group: Take 49 rats the random divides 7 set, 1 mg.kg-1 , 3 mg.kg-1 , 5 mg.kg-1 form namely the normal saline group , model group , fake operation group , compatriot two phosphorus's 0.4 g.kg-1 group (masculine gender contrasts) , Chinese catalpa alcohol. According to document standard, method is eaten.The bureau cooking stove brain lacks for blood rat model preparation: Up to the standard big mice bring into making model range. Adopt right nice oblique tones low-order to open a skull; the electricity curdles to olfactory tract on the right side and the arterial, the preparation permanency bureau cooking stove sex brain lack for the blood model under the cerebrum among a segment of cerebrum between the veins.Catalpa plays a game the cooking stove brain lacks for blood rat nerve behavior effect: Abdominal cavity injection catalpa, compatriot two phosphorus's gallbladders alkali and equivalent menstrual normal saline. The model group and the fake operation group rat do not give medicine. To medicine time, from making a model, 24 h begin, continue giving medicine 7 day. The function grades , the pallet school assignment box tests , the overshot testing box tests , the balance beam testing method is in progress according to that every what be expounded behavior method testing time adopt Bederson nerve previously respectively to every group rat to appraise. Use telecamera itinerary rat behavior, to import a computer, analyses each rat behavior testing evaluation of result, different dosages pure pesticide effect of Chinese catalpa.Pathological mechanism microanatomy HE dyeing is observed:Behavior tests 21st day being finished, the irrigation stream is fixed; the broken end fetches the brain, makes paraffin's section, and is used for HE. 6. Effect catalpa is expressed to newborn mature blood vessel sign egg white Ang-1 and acceptor Tie-2The effect 21st day behavior testing expressing is finished , the irrigation stream is fixed , the broken end fetches the brain , the group who makes paraffin's section , is used for immunity spends detecting Ang-1 , Tie-2 to newborn mature blood vessel sign egg white Ang-1 and acceptor Tie-2. Detecting method draws materials strictly according to Western blot, adopt Western blot method to check infarction cooking stove vicinity cerebral cortex middle Ang-1 egg white's expressing amounts.Results:1. Valuation the Tray taskting box validityIn case that operation forms rat skill last 2, 4, 7, 15, 21 d, grab success rate maintaining in 70.3%±4.1%～72.0%±3.4%has no statistics difference (P> 0.05). The model group , normal saline group , compatriot two phosphorus's gallbladders alkali group come down obviously in creating model last 2nd d , grabbing success rate , come down for 1.8 %±1.3 %～1.8 %±1.7 %, has no statistics difference (P> 0.05). The overshot model group success rate in skill last 2 , 4 , 7 , 15 , 21d comes down to some extent comparing with model group base line value , difference is notable (P<0.05), rat the fore limb restores degree by self in 21d inner are put into much trouble for making that the model is front 35%. The overshot compatriot two phosphorus's gallbladders alkali group success rate in 4 , 7 , 15 , 21 d rises to some extent respectively comparing with model group , difference is notable (P<0.05), the degree it's 21 days' are restored reaches to make model front 44%. Point out that the pallet school assignment box can have the effect reflects a rat being put into much trouble the fore limb is horizontal the tier is a small van taxi overshot capability.2. Valuation the Grasping test box validityGrasping test box reflecting rat being put into much trouble overshot ability that the fore limb perpendicularity face tier. Grab success rate having minor range to undulate, maintain in 70.2%±5.0%～72.1%±4.4%in case that operation forms skill last 2 , 4 , 7 , 15 , 21d, has no statistics difference (P> 0.05). The model group , normal saline group , compatriot two phosphoruses gallbladders alkali group come down obviously in creating model last 2nd d , grabbing success rate , come down for 1.9%±1.9% has no statistics difference (P> 0.05). Model group overshot success rate parting from the regular group , fake operation group , 2 , 4 , 7 , 15 , 21d for in skill last 2 , 4 , 7 , 15 , 21 overshot d success rate has obvious difference comparatively (P <0.05). The overshot model group success rate in skill last 2 , 4 , 7 , 15 , 21d has notable difference comparing with model group base line value (P<0.05), rat the fore limb restores degree by self in 21d inner are put into much trouble being to make that the model is front 32% only. The compatriot two phosphoruses gallbladders alkali forms in 4 , overshot 7 , 15 , 21 d success rate has respectively comparing with model group notable difference (P<0.05), the degree it's 21 days' are restored reaches to make model front 49%.3. Catalpa plays a game the cooking stove brain lacks for blood rat nerve behavior effectBederson scores:Skill last 15 , 21 d, 3mg.kg-1and 5mg.kg-1 form catalpa than the model group , normal saline group and other group gallbladders alkali experimental group recover more well , difference has notable (P<0.05).Tray task test : In case that operation forms rat skill last 2 , 4 , 7 , 15 , 21 d, level overshot success rate is maintained in the base line, undulate in 66.3%±7.5%～70.6%±5.5%, there be no statistics difference (P> 0.05). The model group, normal saline group, catalpa cure the group and compatriot two phosphorus's gallbladders alkali group skill last 2 days , overshot success rate of rat comes down obviously , every overshot group success rate descends for 2.4%±2.2%, group difference has no notable (P> 0.05). Skill last d , every 4th overshot group fore limb function begins to recover , grab success rate rising by 13.8%±4.4%, the group has no obvious difference (P> 0.05). Skill last d , every 7 experimental group rats are put into much trouble overshot success rate of fore limb rises obviously , the model is 15.2%±4.5%set 1mg.kg-1,3mg.kg-1,5mg.kg-1form catalpa respectively for 19.1%±5.6% , 30.2%±7.3%,24.2%±6.6%, 3mg.kg-1,5mg.kg-1 form catalpa among them comparing with model group , difference has notable (P <0.05). Skill last d , every 15 overshot group rat left fore limb functions are still recovering , 3mg.kg-1,5mg.kg-1 form catalpa being compared to the model group's obviously restoring degree highly , difference has notable (P <0.05). Skill last 21 days, the model group are put into much trouble fore limb grab success rate recovering till 25.2%±4.2%, appointment to be equal to base line value 37%; Chinese catalpa alcohol 3mg.kg-1group are 42.8%±4.1%, appointment to be base line value 65%; Chinese catalpa alcohol 5mg.kg-1group are 43.9%±5.3%, appointment to be base line value 66%, difference has notable (P <0.05).Grasping test box:In case that operation forms a rat 2 , 4 , 7 , 15 , 21 d , the level overshot success rate is maintained in the base line, be67.1%±5.0%～69.5%±3.1%. The model group , normal saline group , Chinese catalpa alcohol cure the group and compatriot two phosphorus's gallbladders alkali group skill last 1st d , overshot success rate of rat comes down obviously , descend for 1.7%±1.5%～1.8%±1.4%, group difference has no notable (P> 0.05). Skill last 4th day , every overshot group left fore limb function begins to recover , grab success rate rising by 12.9%±3.2%～15.1%±5.1%, the group has no obvious difference (P> 0.05). The skill queen 7th day , every experimental group rat are put into much trouble overshot success rate of fore limb is rise obviously , among them, the model is 15.5%±2.0%, 1mg.kg-1,3mg.kg-1,5mg.kg-1form catalpa respectively for 20.2%±5.0%,26.8%±4.1%,30.7%±3.4%, cuts blocks for printing pure 1mg.kg-1,3mg.kg-1,5mg.kg-1 group and model group difference have notable (P <0.05). 1mg.kg-1,3mg.kg-1,5mg.kg-1 form skill last 15 days , Chinese catalpa alcohol restoring degree overtopping the model group obviously , difference has notable (P<0.05). Skill last 21 days , the model group are put into much trouble fore limb grab success rate recovering till26.1%±1.1%, appointment to be base line value 39%, the catalpa 1mg.kg-1 group is 46.7%±2.2%, appointment to be base line value 68%, catalpa 3mg.kg-1 group are 50.6%±1.2%, appointment to be base line value 75%, catalpa 5mg.kg-1 group are 52.1%±2.7%, be base line value 76% about , have notable difference comparing with the model (P <0.05).Balance beam test: Skill last 15 days and 21 days, 3mg.kg-1,5mg.kg-1 form catalpa scoring being lower than the model , fake operation , normal saline , compatriot two phosphorus's gallbladders alkali obviously every experimental group, difference has notable (P <0.05).4. Pathological mechanism microanatomy HE dyeing: That the fake operation group rat nerve cell form has no brightness under the mirror, is abnormal , cell membrane , the nuclear membrane are limpid , the nucleolus is obvious. Incomplete blood of model group rat infarction cooking stove brain organization is loose , vacuole appearance unexpected turn of events is obvious , the nerve cell is strengthened shrinking assuming a triangle, cytoplasm is addicted to hashes really , the cell nucleus is strengthened shrinking depths acquire , examine a benevolence to disappear. Lack for blood centre nerve basic escaping from loss the surviving cell has no the entire cell structure obviously, much.5. Catalpa plays a game the cooking stove brain lacks for blood rat Ang-1 , Tie-2 egg white effect expressingAng-1 lacks for the blood rat in the bureau cooking stove brain hitting the target respectively set expressing change: 21st d , IOD of fake operation group rat cerebral cortex Ang-1 egg white value are 16.25±0.06, form 87.77±0.17 with the model, the normal saline forms 112.81±0.11 comparison , have statistics difference (P <0.05). 1mg.kg-1,3mg.kg-1,5mg.kg-1group Ang-1 egg white are worth catalpa respectively for 131.62±0.13,163.77±0.21,172.81±0.33,have notable difference without exception comparing with the model group , normal saline group and compatriot two phosphorus's gallbladders alkali group (P <0.05).Tie-2 lacks for the blood rat in the bureau cooking stove brain hitting the target respectively set expressing change: 21st d , IOD of fake operation group rat cerebral cortex Tie-2 egg white value are 15.20±0.08, form82.81±0.05 with the model, the normal saline forms121.88±0.15comparison , have notable difference (P <0.05). 1mg.kg-1,3mg.kg-1,5mg.kg-1 group Tie-2 egg white are worth catalpa respectively 133.76±0.14,168.79±0.07,173.85±0.22, have notable difference without exception comparing with the model group , normal saline group and compatriot two phosphorus's gallbladders alkali group (P<0.05).Ang-1 change lacking for the blood rat infarction cooking stove vicinity expressing amounts in the bureau cooking stove brain: 21st d , catalpa pure 1mg.kg-1,3mg.kg-1,5mg.kg-1form Ang-1 egg white expressing amounts obvious(P<0.05), have statistics difference comparing with compatriot two phosphorus's gallbladders alkali group (P <0.05). The model group , normal saline Ang-1 egg white of group express amounts difference and have difference(P<0.05), this changes a trend and the immune group of Ang-1 spends detecting result to coincide each other.Conclusion:1.The pallet school assignment box and overshot testing box experiment method are stable , effective, can be put into much trouble to brain apoplexy rat meticulous action of overshot fore limb melon seed makes objective valuation , may be the experiment technology platform observing appraising simple limb obstacle degree and their medicine effect objectively.2. Bederson nerve functions grade, overshot success rate of pallet school assignment box measures judge, grab the testing box to grab success rate measure judge, balance beam go on foot success rate tests, have included the nerve symptom, minute motion is in harmony, feeling motion integrates, behavior tests an index from simplicity to complicated nerve, may test system as incomplete brain blood rat behavior, is used to get ready for construction of the behave test systems. 3. Catalpa can improve the bureau cooking stove nature brain lacking for the blood rat Bederson nerve function grading, the pallet school assignment box measures the overshot success rate judging, grab the success rate testing a box measuring overshot success rate, balance beam judging going on foot testing, play a game the cooking stove nature brain lacks for functional recovering of blood rat limb motion have notable promote an effect.4. Catalpa moves Ang-1 and their acceptor Tie-2 up, boosts the bureau cooking stove nature brain lacking for blood rat freshman blood vessel maturity.
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