鞘内注射PKC抑制剂对神经病理痛大鼠脊髓背角NMDAR、CGRP表达的影响木

目的:观察鞘内注射PKC抑制剂对神经病理痛大鼠脊髓背角N-甲基-D-天冬氨受体(NMDAR)、降钙素基因相关肽(CGRP)表达的影响.方法:健康雄性SD大鼠36只,体重220～280g.随机分为4组(n=9):对照组(C组)、假手术组(Sham组)、坐骨神经分支选择结扎切断模型组(SNI组)和PKC抑制剂组(P组).SNI组和P组制备SNI模型,P组在SNI术后14d内每天鞘内注射PKC抑制剂11μg,其余各组给予等容量生理盐水.在SNI术前ld(基础值)及术后14d每次注射药物后测定机械痛阈和热缩足潜伏期,分别于SNI术后2、7、14d注射药物后各处死大鼠3只,采用免疫组化法测定L5节段脊髓背角NMDAR和CGRP表达水平.结果:与C组和Sham组比较,SNI组机械痛阈降低,NMDAR和CGRP表达上调(P<0.01),热缩足潜伏期差异无统计学意义(P>0.05).与SNI组比较,P组机械痛阈升提高,热缩足潜伏期延长,NMDAR和CGRP表达下调(P<0.01).结论:鞘内注射PKC抑制剂对神经病理痛大鼠具有明显的抗伤害效应,并抑制大鼠脊髓背角NMDAR和CGRP表达.     

鞘内注射PKC抑制剂对神经病理痛大鼠脊髓背角NMDAR、CGRP表达的影响

目的：观察鞘内注射PKC抑制剂对神经病理痛大鼠脊髓背角N-甲基-D-天冬氨受体（NMDAR）、降钙素基因相关肽（CGRP）表达的影响。方法：健康雄性SD大鼠36只,体重220～280g。随机分为4组（n=9）：对照组（C组）、假手术组（Sham组）、坐骨神经分支选择结扎切断模型组（SNI组）和PKC抑制剂组（P组）。SNI组和P组制备SNI模型,P组在SNI术后14d内每天鞘内注射PKC抑制剂11μg,其余各组给予等容量生理盐水。在SNI术前1d（基础值）及术后14d每次注射药物后测定机械痛阈和热缩足潜伏期,分别于SNI术后2、7、14d注射药物后各处死大鼠3只,采用免疫组化法测定L5节段脊髓背角NMDAR和CGRP表达水平。结果：与C组和Sham组比较,SNI组机械痛阈降低,NMDAR和CGRP表达上调（P〈0.01）,热缩足潜伏期差异无统计学意义（P〉0.05）。与SNI组比较,P组机械痛阈升提高,热缩足潜伏期延长,NMDAR和CGRP表达下调（P〈0.01）。结论：鞘内注射PKC抑制剂对神经病理痛大鼠具有明显的抗伤害效应,并抑制大鼠脊髓背角NMDAR和CGRP表达。     

鞘内维拉帕米对甲醛致痛大鼠的抗伤害作用

目的观察鞘内注射维拉帕米( Ver)后大鼠伤害性行为学反应,脊髓背角 Fos表达的改变,从组织化学及行为学角度探讨维拉帕米对甲醛致痛大鼠的抗伤害作用的可能机制. 方法雄性 SD大鼠 24只,随机数字表法分为 3组 生理盐水对照组( NS组) , 甲醛对照组 (F组 )和维拉帕米甲醛组( VeF组). F组给予 50 mL/L甲醛 100 μ L,足底注射, VeF组在同 F组处理前鞘内给予 Ver.在甲醛处理后观察大鼠的行为学表现并检测大鼠脊髓 Fos的表达. 结果 VeF组的缩腿舔爪时间、脊髓的 Fos表达显著短于或弱于 F组. 结论 Ver能抑制大鼠脊髓背角 Fos的释放,可能是其产生抗伤害作用的机制之一.     

鞘内单次注射吗啡对神经病理痛大鼠脊髓背角CGRP表达的影响

目的:观察鞘内单次注射吗啡对神经病理痛大鼠脊髓背角降钙素基因相关肽(Calcitonin gene-related peptide CGRP)表达的影响.方法:20只健康雄性SD大鼠,体重200～270 g,随机分为4组:对照组(C组,n=5),假手术组(S组,n=5),生理盐水组(N组,n=5),吗啡组(M组,n=5).C组不做任何处理,其它3组均根据改良Yaksh法进行鞘内置管;N组和M组制备神经病理痛模型(SNI模型),制模2 d后,N组和M组分别鞘内注射生理盐水和吗啡10μg,容量为20μl.每组均在SNI术前1d(基础值)到术后2d内每天测定机械痛阈和热缩足潜伏期,并全部在注药后2h处死大鼠,用免疫组织化学方法观察大鼠L5节段水平脊髓背角CGRP的表达.结果:与C组和S组比较,N组机械痛阈降低,CGRP表达上调(P<0.05),热缩足潜伏期差异无统计学意义(P0.05).与N组比较,M组机械痛阈升高,热缩足潜伏期延长,CGRP表达下调(P<0.05).结论:鞘内单次注射吗啡对神经病理痛大鼠在产生镇痛效应时引起脊髓背角CGRP表达下调.     

鞘内维拉帕米对甲醛致痛大鼠的抗伤害作用

目的：观察鞘内注射维拉帕米(Ver)后大鼠伤害性行为学反应，脊髓背角Fos表达的改变，从组织化学及行为学角度探讨维拉帕米对甲醛致痛大鼠的抗伤害作用的可能机制。方法：雄性SD大鼠24只，随机数字表法分为3组：生理盐水对照组(NS组)，甲醛对照组(F组)和维拉帕米甲醛组(VeF组)。F组给予50mL／L甲醛100μL，足底注射，VeF组在同F组处理前鞘内给予Ver。在甲醛处理后观察大鼠的行为学表现并检测大鼠脊髓Fos的表达。结果：VeF组的缩腿舔爪时间、脊髓的Fos表达显著短于或弱于F组。结论：Ver能抑制大鼠脊髓背角Fos的释放，可能是其产生抗伤害作用的机制之一。     

前炎性细胞因子白细胞介素1β和肿瘤坏死因子在保留神经损伤所致机械性痛敏发生中的不同作用

背景脊髓胶质细胞的激活参与了保留神经损伤(spared nerve injury,SNI)诱致的机械性痛敏的发生.脊髓胶质细胞激活释放的前炎性细胞因子在这个过程中是否扮演了重要的角色.目的探讨了白细胞介素1(interleukin-1,IL-1)和肿瘤坏死因子(tumor necrosis factor,TNF)在SNI诱致的机械性痛敏中的不同作用.设计随机对照试验.地点和对象解放军第四军医大学实验动物中心提供的40只健康成年雄性Sprague-Dawley大鼠.干预实验分两组实验组鞘内分别给予白细胞介素-1受体拮抗剂(interleukin-1 receptor antagonist,IL-1ra),肿瘤坏死因子抗血清(tumornecrosisfactor anti-serum,anti-TNF)和同时给予IL-1ra和anti-TNF.对照组鞘内给予相应的溶媒.术前和术后分别检测大鼠后爪机械性刺激阈值.主要观察指标大鼠检测机械性刺激缩足阈值.结果①鞘内给予IL-1ra可以对大鼠单侧后爪内侧和外侧在术后1周内产生一个明显的对机械性痛敏的抑制,而anti-TNF没有这样的作用.②同时鞘内给予IL-1ra和anti-TNF可以产生更加明显的机械性痛敏抑制,抑制作用可以长达术后6周.结论脊髓胶质细胞激活释放的前炎性细胞因子IL-1和TNF在SNI诱致的机械性痛敏中扮演了重要的角色.IL-1和TNF在SNI诱致的机械性痛敏中的作用不同.     

鞘内注射AIP对神经病理性痛大鼠的镇痛作用及脊髓背角神经元磷酸化CREB的影响

目的:探讨钙/钙调素依赖性蛋白激酶Ⅱ(calcium/calmodulin-dependent protein kinaseⅡ,CaMKⅡ)在神经病理性疼痛中的作用。方法:坐骨神经选择性损伤(SNI)大鼠模型,随机分为4组(n=8):SNI组,Sham组,NS组,AIP组,后两组分别于术前20 min鞘内注射10μl的生理盐水或10%的AIP。于术后1d、2d、3d、4d、6d、8d、10d、14d测定大鼠机械缩足反射阈值(mechanical with-drawal threshold,MWT)。分别于术后1d、3d、7d取大鼠脊髓腰段,用免疫组织化学法(n=6)检测脊髓背角磷酸化pCaMKⅡ的表达;Western blot法(n=4)检测脊髓背角神经元细胞核内pCREB的表达。结果:术后1~3d AIP组大鼠MWT较NS组明显升高(P<0.01);术前鞘内注射AIP在术后1d、3d能够使脊髓背角pCaMKⅡ的表达减少,同时脊髓背角神经元细胞核内pCREB的表达也明显减少。结论:CaMKⅡ参与了神经病理性痛的形成,其作用部分通过CREB介导。     

前炎性细胞因子白细胞介素1β和肿瘤坏死因子在保留神经损伤所致机械性痛敏发生中的不同作用

背景：脊髓胶质细胞的激活参与了保留神经损伤(spared nerve injury.SNI)诱致的机械性痛敏的发生。脊髓胶质细胞激活释放的前炎性细胞因子在这个过程中是否扮演了重要的角色。目的：探讨了白细胞介素1(interleukin-1，IL-1)和肿瘤坏死因子(tumor necrosis factor，TNF)在SNI诱致的机械性痛敏中的不同作用。设计：随机对照试验。地点和对象：解放军第四军医大学实验动物中心提供的40只健康成年雄性Sprague-Dawlev大鼠.干预：实验分两组：实验组：鞘内分别给予白细胞介素-1受体拮抗剂(interleukin-1 receptor antagonist，IL-1ra)．肿瘤坏死因子抗血清(tumor necrosis factor anti-serum，anti-TNF)和同时给予IL-1ra和anti-TNF。对照组：鞘内给予相应的溶媒。术前和术后分别检测大鼠后爪机械性刺激阈值。主要观察指标：大鼠检测机械性刺激缩足阈值。结果：①鞘内给予IL-1ra可以对大鼠单侧后爪内侧和外侧在术后1周内产生一个明显的对机械性痛敏的抑制，而anti-TNF没有这样的作用，②同时鞘内给予IL-1ra和anti-TNF可以产生更加明显的机械性痛敏抑制，抑制作用可以长达术后6周。结论：脊髓胶质细胞激活释放的前炎性细胞因子IL-1和TNF在SNI诱致的机械性痛敏中扮演了重要的角色。IL-1和TNF在SNI诱致的机械性痛敏中的作用不同。     

胫骨骨癌痛模型大鼠鞘内注射舒芬太尼后脊髓背角N-甲基-D-天冬氨酸受体及降钙素基因相关肽的表达

背景:临床和动物实验证实舒芬太尼鞘内注射有明显的镇痛效果,但其机制尚不明确。目的:观察鞘内注射舒芬太尼对骨癌痛大鼠痛阈及脊髓背角N-甲基-D-天冬氨酸受体、降钙素基因相关肽表达的影响。方法:健康SD大鼠36只随机等分为对照组、假手术组、模型组和舒芬太尼组。后2组利用乳腺癌细胞腹水制备胫骨骨癌痛模型,假手术组注射加热灭活后的死细胞。模型组和舒芬太尼组在建模后15d内每天鞘内注射生理盐水和舒芬太尼。结果与结论:与对照组和假手术组比较,模型组大鼠在建模后第12和14天脊髓背角N-甲基-D-天冬氨酸受体和降钙素基因相关肽表达增加(P<0.01),且出现机械性异常疼痛痛阈降低,热刺激后爪退缩潜伏时间缩短(P<0.01);而与模型组比较,舒芬太尼组大鼠,在注药后第12和14天脊髓背角浅层N-甲基-D-天冬氨酸受体和降钙素基因相关肽的表达降低(P<0.01),且机械刺激痛阈提高以及热刺激后爪潜伏期延长(P<0.01)。提示鞘内注射舒芬太尼对骨癌痛大鼠具有明显的抗伤害效应,其机制与抑制大鼠脊髓背角N-甲基-D-天冬氨酸受体和降钙素基因相关肽表达有关。     

靶向抑制脊髓5-羟色胺能通路对大鼠骨癌痛的影响

目的:探讨靶向抑制脊髓5-羟色胺能通路对大鼠骨癌痛的影响。方法:雌性SD大鼠70只,体重160~180 g,随机分组。骨癌痛组大鼠于胫骨骨髓腔注射Walker 256大鼠乳腺癌细胞悬液10μl制备癌痛模型。假手术组于骨髓腔注射D-hank's液10μl。大鼠造模后第10天行鞘内置管,第14天鞘内注射生理盐水或5-羟色胺选择性神经毒素5,7-DHT。鞘内给药前1天和给药后5天每天测定机械痛阈。鞘内给药后第3天取脊髓背角组织采用高效液相色谱法检测5-羟色胺含量。大鼠测痛结束后行胫骨X线摄片,观察肿瘤导致的骨破坏程度。结果:X线片显示骨癌痛大鼠术侧胫骨可见肿瘤生长导致的骨皮质大片缺损。与鞘内给予生理盐水的骨癌痛组大鼠相比,骨癌痛组大鼠鞘内给予5,7-DHT后机械性痛觉超敏显著缓解(P<0.05),缩爪持续时间明显缩短(P<0.05)。与鞘内给予等体积生理盐水的大鼠相比,鞘内给予60μg的5,7-DHT可以显著的降低脊髓背角中5-羟色胺含量。结论:本实验证实了鞘内注射5,7-DHT选择性消除脊髓5-羟色胺可显著缓解骨癌痛大鼠的疼痛,说明脊髓背角中5-羟色胺含量的增加参与了大鼠骨癌痛的维持。     

EXPERIMENTS WITH POLIOMYELITIS IN THE RABBIT

The poliomyelitic virus obtained from an experimental monkey has been passed through eight generations in rabbits. It shows no signs of dying out. On the other hand, it gives no evidence of becoming more pathogenic to the species through successive passage. The period of incubation remains variable and the percentage of takes has not increased. Whether eventually a virus can be obtained which is of heightened virulence to rabbits is problematic. All inoculations are by no means successful. The animals show great individual differences in susceptibility to the virus, as is evidenced by the fact that out of fifty-four rabbits inoculated, only twenty-two, or about 40 per cent, succumbed. This fact may explain the negative results of other investigators. At several points in the series of experiments it was thought that the strain had died out. As many as six rabbits have been inoculated one after the other before the virus would catch again. The age of the rabbits is important in considering the susceptibility. From the limited data at our command, adult rabbits are resistant, and there appears to be an abrupt increase in resistance between the age of 6 and 8 weeks; that is, rabbits under 6 weeks are more susceptible to the virus. There seems to be a parallel between the age incidence of this disease in rabbits and spontaneous poliomyelitis in man. The age incidence of poliomyelitis in man is indicated by the term "infantile paralysis." Several methods of inoculation have proved successful; thus the rabbits have succumbed as a result of introducing the virus directly into the brain, by injecting it into a peripheral nerve, or directly into the circulation, or by placing it upon the uninjured nasal mucosa. The symptoms produced show more or less departure from the symptoms of poliomyelitis as seen in the spontaneous disease in man and in the experimental disease in the monkey. There are two distinct pictures recognizable. In one there is paralysis of one or more of the extremities which progresses until death, resembling somewhat the symptoms of the experimental disease in the monkey. This we have designated the progressive type. The other group is included in what we have called the fulminating type. The symptoms are explosive in character, with extreme weakness amounting to prostration, terminating in death in a few hours, attendant upon respiratory failure. The mode of inoculation seems to have little effect upon the type of symptoms produced. The period of incubation is variable and apparently does not depend upon the method of inoculation. The period varied from 2 to 41 days, with an average of 12 days. The two extremes both occurred after intracranial injection. After intranasal insufflation the incubation period was short, being in each case 2 days, followed by symptoms of the fulminating type. The placing of the virus into the nose seems to be an effective method, but is as uncertain as other routes, as only three out of nine rabbits tested in this manner succumbed. The disease produced by this route was particularly virulent. The virus shows no tendency to become fixed. The period of incubation is as variable in the eighth generation as in the first, and the virus has shown no tendency towards increasing virulence through successive passage, in these respects differing from the virus of rabies. We have found the virus to be filterable. An emulsion of the central nervous matter of a rabbit of the first generation passed through a Berkefeld filter, and injected intracerebrally into another rabbit, resulted in death, preceded by symptoms of the fulminating type. Virus (unfiltered) from this rabbit was transferred successfully to two other rabbits. The lesions, while definite and consistent throughout the series, lack the distinctive features of the pathologic picture of poliomyelitis in man and the monkey. Capillary congestion, punctate hemorrhages, degeneration of the motor cells, satellitosis, and more or less cellular infiltration of the gray matter of the cord and medulla are found, but perivascular infiltration is absent and the infiltrating cells are not lymphocytic in character. One of the most striking features of this investigation is the way in which rabbits and monkeys react to the same virus. The disease in the rabbit presents certain clinical resemblances to the experimental disease in the monkey and also to the spontaneous disease in children. On the other hand, the symptoms show marked variation from those seen in the monkey and in man. The picture has not the same constancy in rabbits and could not in most cases be recognized clinically as poliomyelitis. There are still more marked differences in the pathology. While it is true that the brunt of the attack in the rabbit falls upon the gray matter of the cord and medulla, the appearance of the lesions under the microscope shows such differences from the lesions of experimental poliomyelitis in monkeys, as well as the natural disease in man, as to suggest two distinct infections. It is more reasonable, however, to assume that we are dealing with a modified form of poliomyelitis; that the rabbit reacts differently to the virus than the monkey or man; and that the disease produced in rabbits by us and others is in fact poliomyelitis. So far as we know, no other virus produces such differences in two animal species. Smallpox is so profoundly altered in the cow that it took almost 100 years to prove Jenner''s assumption that cowpox is a modified form of smallpox. However, the pock of vaccinia is a correct counterpart both clinically and pathologically of the pock of variola. If the virus of poliomyelitis may be so altered in the rabbit as scarcely to be recognizable, may it not be still more profoundly changed in other animals? The conjecture then arises that poliomyelitis, instead of being limited naturally to man and experimentally to monkeys, may in fact occur in other animals in unnoticed or unrecognized form. If this should prove true, it may be a source of human infection and may help to solve the problem of prevention.     

BILE DUCT HAMARTOMA OCCURRING IN ASSOCIATION WITH LONG-TERM TREATMENT WITH DANAZOL

We report a case of bile duct hamartoma which developed in a patient who had been on long-term danazol treatment. Such patients should be under close follow-up, preferably with periodic ultrasound examination of the liver. If the patient develops a liver mass, because of non-specific clinical features and imaging appearances, biopsy may be the only way to achieve a definitive diagnosis.