孕酮对兔脊髓急性缺血再灌注损伤后细胞凋亡的影响

目的探讨孕酮对兔脊髓急性缺血再灌注损伤后细胞凋亡的影响。方法将90只日本大耳白兔随机分为假手术组(n=30)、脊髓损伤组(n=30)与孕酮治疗组(n=30),各组又进一步分为12 h、24 h、36 h、48 h、72 h、14 d等6个亚组,每亚组5只。假手术组只行腹部切开术但不阻断腹主动脉。脊髓损伤组和孕酮治疗组均制作缺血再灌注模型,制模成功后孕酮治疗组每24h注射孕酮1次(将孕酮溶于玉米油,浓度10 mg/ml,注射剂量为8 ml/kg),脊髓损伤组在相同时间点注射等量生理盐水。采用TUNEl染色评估细胞凋亡,采用BBB评分评估运动功能。结果 HE染色结果显示,假手术组脊髓形态正常;脊髓损伤组可见脊髓结构被破坏,正常神经元数目减少;孕酮治疗组较脊髓损伤组正常神经元数目多。TUNEL染色结果显示,脊髓损伤组和孕酮治疗组各时间点凋亡细胞数均明显高于假手术组(P0.05);孕酮治疗组各时间点凋亡细胞的数目均明显少于脊髓损伤组(P0.05)。假手术组术后12 h BBB评分为17.3分,术后14 d时升高到21.0分;脊髓损伤组术后12 h BBB评分为0.6分,随后缓慢升高,术后14 d时评分为9.2分。孕酮治疗组BBB评分变化与脊髓损伤组相似,但术后36 h开始,各时间点评分均明显高于相应脊髓损伤组(P0.05)。结论孕酮治疗可改善脊髓急性缺血再灌注损伤兔运动功能,可能与抑制细胞凋亡有关。     

尼莫地平对脑缺血早期神经元型一氧化氮合酶的影响

目的观察局灶性脑缺血大鼠早期缺血部位神经元型一氧化氮合酶(neuronal nitric oxide synthase,nNOS)的变化,以及钙拮抗剂尼莫地平对其的影响。方法以线栓法制作大鼠大脑中动脉闭塞模型为基础,选取雄性SD大鼠共75只,随机分成假手术组(n=25)、模型对照组(n=25)和尼莫地平处理组(n=25)。以NDAPH-d组织化学方法观测各组大鼠脑缺血后30min、2h和6h纹状体中nNOS阳性细胞的变化;利用生物化学法和Hoechst凋亡染色分别检测各组大鼠脑缺血30min纹状体nNOS活性和6h半暗区细胞凋亡。结果脑缺血30min,与假手术组相比,模型对照组大鼠缺血侧纹状体中nNOS活性和nNOS阳性细胞数均升高(P<0.01),而尼莫地平组无明显变化(P>0.05);随脑缺血时间延长,模型对照组和尼莫地平组nNOS阳性细胞数都下降(P<0.05);脑缺血6h,尼莫地平组半暗区的凋亡细胞少于模型对照组(P<0.01)。结论钙拮抗剂尼莫地平可抑制脑缺血早期的nNOS上升,并减轻随后的脑组织损伤。     

神经干细胞动脉移植治疗脊髓损伤后Bcl-2与Bax的变化

目的建立肾下腹主动脉输注神经干细胞(neural stem cell,NSC)治疗脊髓缺血损伤(spinal cord ischemia injury,SCII)大鼠模型,探讨神经干细胞移植治疗脊髓缺血损伤的机制。方法 54只成年SD雌性大鼠随机分为假手术组、对照组和NSC组,每组18只。假手术组进行手术操作,不阻断动脉;对照组大鼠行脊髓缺血再灌注损伤模型,阻断肾下腹主动脉120 min后开放;NSC组完成SCII操作后向留置针缓慢推注体外培养同源NSC100万。每组取5只动物分别于术后5 d、10 d、15 d行BBB评分和运动诱发电位检测,余下每组13只大鼠于术后7 d取材,其中8只动物用于RT-PCR和Western blot检测,5只动物用于Tunnel染色。结果术后7 d,TUNEL染色发现对照组和NSC组L2节段均存在大量凋亡细胞;术后7 d对照组和NSC组Bax和Bcl-2蛋白表达水平较假手术组明显增加(P<0.05),其中NSC组Bax蛋白表达水平低于对照组(P<0.05),Bcl-2蛋白表达水平高于对照组(P<0.05);NSC组Bcl-2/Bax蛋白质表达比值较对照组增加(P<0.05)。BBB评分和MEP显示对照组和NSC组大鼠术后均存在显著功能障碍,其中NSC组大鼠BBB评分于术后10¹5 d高于对照组(P<0.05),且MEP(10 d)的波幅增大(P<0.05)而潜伏期缩短(P<0.05)。结论经股动脉移植NSC,可通过上调Bcl-2表达和下调Bax表达而抑制缺血再灌注损伤脊髓细胞凋亡的发生,从而促进动物后肢运动功能的恢复。     

动脉移植BMSCs对大鼠缺血再灌注损伤脊髓CNTF和STAT3的影响

目的探讨肾下腹主动脉移植骨髓间充质干细胞(BMSCs)对大鼠缺血再灌注损伤脊髓CNTF和STAT3的影响及对损伤脊髓功能恢复的作用。方法 24只成年SD雌性大鼠随机分为假手术组、对照组及移植组3组,每组8只。术后对大鼠进行后肢神经行为学评分和运动诱发电位检测,采用RT-PCR、Western blot检测大鼠缺血节段脊髓内CNTF和STAT3的表达变化。结果与假手术组比较,对照组和移植组BBB评分于术后0 d、5 d、10 d、15 d显著降低(P<0.01),MEP潜伏期延长(P<0.01)、波幅减小(P<0.01),脊髓CNTF和STAT3 mRNA和蛋白表达增加(P<0.01);与对照组比较,移植组BBB评分于术后5¹5 d增高(P<0.01),MEP潜伏期缩短(P<0.01)、波幅增加(P<0.01),CNTF和STAT3 mRNA和蛋白表达增加(P<0.01)。结论肾下腹主动脉移植BMSC可能通过增加损伤脊髓局部CNTF和STAT-3表达促进脊髓缺血再灌注损伤大鼠后肢功能恢复。     

丹红注射液对脑缺血再灌注损伤大鼠ICAM-1表达的影响

目的 探讨丹红注射液对脑缺血再灌注损伤大鼠ICAM-1表达和白细胞浸润的影响.方法 SD大鼠24只,随机分为假手术组(n=8)、缺血再灌注组(n=8)和bFGF组(n=8).应用线栓法制作大鼠局灶性脑缺血再灌注模型,大脑中动脉阻塞1 h再灌注损伤24 h,术前治疗组丹红注射液预处理3 d,假手术组及缺血再灌注组用生理盐水预处理,采用免疫组织化学法染色和组织HE染色检测缺血区脑微血管内皮ICAM-1的表达及白细胞计数.结果 缺血再灌注组与假手术组相比ICAM-1表达明显升高,白细胞浸润明显(P<0.05).丹红治疗组ICAM-1表达较缺血再灌注组明显减少,白细胞浸润程度减轻(P<0.05).结论 丹红注射液预处理可降低局灶性脑缺血再灌注损伤大鼠ICAM-1表达,减轻白细胞浸润,提示抑制粒细胞黏附作用是其脑保护作用机制之一.     

腺苷预处理对大鼠脊髓缺血再灌注损伤保护作用的实验研究

目的探讨腺苷预处理对脊髓缺血再灌注损伤的保护作用。方法制作大鼠脊髓缺血再灌注损伤模型。72只SD大鼠随机分为3组:假手术组(A组)、缺血再灌注组(B组)、腺苷预处理组(C组),再按缺血再灌注后不同时间把各组随机分成3个亚组。通过BBB神经评分观察3组大鼠的神经症状;应用HE染色评价脊髓内的病理组织学变化;并测定3组大鼠不同时间点的SOD、MDA含量。结果 A组、C组在各个时间点的BBB评分明显高于B组(P<0.05);与B组相比,A组、C组细胞形态学变化轻,脊髓组织的SOD含量明显增高(P<0.05),MDA明显降低(P<0.05)。结论腺苷预处理通过提高SOD水平,降低氧自由基和脂质过氧化对大鼠脊髓缺血再灌注损伤进行保护,使神经损伤症状得到一定程度的恢复。     

IGF-1对大鼠局灶性脑损伤c-fos表达的影响

目的研究胰岛素样生长因子-1(IGF-1)对大鼠局灶性脑缺血再灌注后c-fos表达的影响及与缺血时间关系,探讨IGF-1对脑缺血再灌注损伤的保护作用。方法制作SD大鼠大脑中动脉缺血再灌注模型。将55只SD雄性大鼠随机分为假手术组(n=5)、对照组(n=25)、IGF-1治疗组(n=25),其中后2组按缺血再灌时间(6h、12h、1d、3d、7d)不同可分为5个亚组,每组5只,治疗组于缺血2h再灌注1h后经腹腔注入40μg/kg稀释为1 ml的IGF-1,假手术组及对照组同时腹腔注入生理盐水1 ml。以上动物均在再灌注后规定时间点用4%多聚甲醛经心脏灌注固定,取大鼠脑组织,应用免疫组化S-P法和HE染色检测c-fos蛋白表达及脑组织结构病理变化。结果与对照组相比,治疗组大鼠脑组织c-fos表达明显减少,神经细胞坏死程度明显减轻。结论IGF-1在大鼠局灶性脑缺血再灌注损伤中起保护作用,其作用机制包括降低c-fos的表达,发挥神经保护作用。     

活性氧在脑缺血-再灌注损伤后动态表达

目的观察活性氧(ROS)在大鼠脑缺血—再灌注损伤不同时间缺血灶周区皮质的动态表达。方法健康成年SD雄性大鼠(n=55),随机分成假手术组(n=5)和模型组(n=50),模型组按再灌注时间分为10个组,即脑缺血—再灌注1h、3h、6h、12h、1d、3d、5d、7d、14d和28d组,每组5只大鼠。通过阻塞大脑中动脉(MCAO)制作大鼠局灶性脑缺血—再灌注损伤模型,采用改良神经功能严重性评分(m NSS)对不同时间点大鼠的神经功能进行评价,采用流式细胞术检测缺血灶周区ROS和血管内皮生长因子(VEGF)动态时空表达。结果假手术组未见神经功能损伤,评分为0。脑缺血—再灌注1h大鼠出现神经损伤,3h~1d时症状加重,3~7d时症状有所改善,14~28d症状明显好转。假手术组有少量ROS和VEGF表达,脑缺血再灌注损伤后1h缺血灶周区ROS急剧上升,3~6h有所下降,但仍远高于假手术组,后逐渐下降,3d时降到最低,与假手术组相仿,随后迅速上升,5d时已超过6h数值,7d时接近1h高度,后ROS表达呈平台期,VEGF表达趋势与其相仿,ROS表达与神经功能评分呈负相关。结论 ROS可能在脑缺血损伤中起双相作用,在超早期参与脑损伤过程,后期可能参与脑修复作用。     

尼莫地平对大鼠急性脑缺血再灌注损伤早期保护作用的研究

目的探讨尼莫地平对急性脑缺血再灌注损伤的早期保护作用。方法线栓法复制大鼠急性脑缺血模型。30只雄性Wistar大鼠随机分为假手术、模型、尼莫地平3组。模型组采取缺血2h再灌注2h。尼莫地平组大鼠在缺血0时刻起每小时腹腔注射给药一次,剂量为5mg/kg。各组大鼠在手术4h后实验结束后,行腹主动脉采血,同时取完整脑组织。脑组织切片进行TTC染色,并比较各组脑组织梗死面积。检测各组大鼠血清及脑组织匀浆中SOD、MDA、NO含量。结果与模型组相比,尼莫地平组大鼠脑组织梗死面积显著减少。血清生化指标显示,模型组SOD含量显著低于假手术组,给予尼莫地平治疗后,SOD含量增加明显。模型组MDA、NO含量明显高于假手术组,尼莫地平组明显降低血清中MDA、NO。结论尼莫地平对大鼠急性脑缺血再灌注损伤有保护作用,这种保护作用与NO和氧化应激密切相关。     

异丙酚及缺血预处理联合对主动脉阻断致脊髓损伤的保护

目的 探讨缺血预处理(ischemic preconditioning,IPC)和异丙酚（Propofol）预处理对兔主动脉阻断所致脊髓损伤的防治作用其可能作用机制。 方法 本实验有二处理因素：缺血预处理及异丙酚预处理。采用2×2析因实验设计,分四个实验组：缺血再灌注损伤组（A组,空白组）、缺血预处理组（B组）、异丙酚组（C组）及缺血预处理和异丙酚联合预处理组（D组）。雄性日本大白兔32只,随机分为4组,每组8只。各组阻断腹主动脉40min,再灌注7d。B组IPC5min,再灌注30min后阻断腹主动脉40min,再灌注7d;C组静注异丙酚5mg/kg 10min后阻断腹主动脉40min,再灌注7d;D组IPC5min,,再灌注20min时静注异丙酚5mg/kg,再灌注30min时阻断腹主动脉血流40分钟,再灌注7天。分别测定阻断前10min（C-10）、开放前即刻（C40）、再灌注60min（R60）及7d（R7d）血清MDA、SOD;观察术后后肢神经功能;脊髓病理学观察;脊髓凋亡神经元。 结果 ①缺血再灌注后B、C、D组MDA值明显高于C-10值及A组相应时点值（p<0.05）, SOD值变化同MDA变化相反（p<0.05）; B组缺血后各时点MDA值明显低于C组(P<0.05), D组缺血后各时点MDA值明显低于B、C组(P<0.05),SOD值变化同MDA变化相反（p<0.05）。②B、C、D组凋亡细胞数明显少于A组(P<0.05); B组明显少于C组(P<0.05),D组明显少于B、C组(P<0.05)。③B、C、D组瘫痪发生率明显低于A组(P<0.05),B组瘫痪发生率明显低于C组(P<0.05),D组瘫痪发生率明显低于B、C组(P<0.05);B、C、D组后肢神经功能评分明显高于A组(P<0.05), B组后肢神经功能评分高于C组(P<0.05),D后肢神经功能评分高于B、C组(P<0.05)。④ D组脊髓病理变化明显轻于A、B、C组(P<0.05)。结论 缺血预处理和异丙酚预处理对兔主动脉阻断所致脊髓损伤都有良好的防治作用;缺血预处理对脊髓缺血再灌注损伤的作用明显好于异丙酚;缺血预处理和异丙酚联合应用对脊髓缺血再灌注损伤有更加良好的防治作用,表现出一定的交互作用。缺血预处理和异丙酚联合预处理对兔主动脉阻断所致脊髓损伤其机制可能与其抗氧化反应作用有关。     

Mechanisms of the suppression of the nociceptive reflex of the opening of the mouth during stimulation of the structures of the limbic brain

The comparative effectiveness of the inhibitory influence of tetanic stimulation of hypothalamus, amygdala and limbic cortex on EMG-response of m. digastricus evoked by electrical stimulation of tooth pulp nociceptive afferents was studied in cats anesthetized with a mixture of chloralose and nembutal. It was found that inhibition of the EMG-component of the jaw-opening reflex is most pronounced in case of stimulation of medial and lateral region of the hypothalamus, the inhibitory effect of central and medial nuclei of the amygdala is less pronounced and the effect of the limbic cortex is the weakest. It was shown that the mechanism of the antinociceptive effect of tetanic stimulation of the hypothalamus is not related to the concomitant increase of the blood pressure. After stabilization of the blood pressure the suppressive effect of the hypothalamus remains without changes, that points out to a direct, primary, not baro-afferent mechanism of the inhibition of the activity of nociceptive neurons of the trigeminal sensory nuclei. Noradrenaline, injected intravenously, induced a large increase of the blood pressure accompanied by a pronounced inhibition of the pain reflex. Angiotensin causes the same degree of blood pressure elevation without changes in the amplitude of the EMG-response of the pain reflex. Hypothalamic and noradrenergic mechanisms for control of pain sensitivity are discussed.     

药物治疗与合并认知行为治疗对强迫症疗效的比较

目的探讨认知行为心理治疗(CBT)在强迫症(OCD)患者各亚型治疗中的有效性和规律性。方法本研究为临床对照研究。符合入组标准的强迫症患者按患者自愿原则分为两组,治疗观察3、6、12个月。疗效评定分别运用Yale-Brown强迫量表,自拟的自评好转程度量表和临床疗效评定。结果认知行为心理治疗合并药物治疗组31例,临床有效率70.9%,其中治愈率1.8%。单纯药物治疗组24例,临床有效率33.3%。Yale-Brown强迫量表和自评量表得分在6个月和12个月两组有显著差异(P<0.05)。其中强迫症亚型(怕脏型、反复检查型和反复担心型)的疗效比较,怕脏型在治疗3个月末两组间自评量表评分有显著性差异(P<0.05);反复担心型在治疗6个月末两组间Yale-Brown强迫量表总分有显著性差异(P<0.05);反复检查型两组间无统计学差异。结论认知行为心理治疗合并药物治疗强迫症的疗效明显优于单纯药物治疗。强迫症的亚型在治疗中的有效性次序为:反复担心型>怕脏型>反复检查型。     

Characteristics of the sympathetic innervation of the nictitating membrane and of the vasculature of the nose and tongue of the cat

Summary Vasomotor responses from the nasal mucosa and tongue, and contractions of the nictitating membrane, were recorded on stimulation of the cervical sympathetic or internal carotid nerves.Preganglionic sympathetic nerve fibres which elicited a membrane response possessed a lower threshold than those which evoked nasal vasoconstriction, while the latter displayed a lower threshold than fibres which evoked tongue vasoconstriction. The sympathetic vasodilator fibres to the tongue, whose activity was revealed after-receptor blockade, had a similar threshold to the vasoconstrictor fibres.Membrane contraction, nasal vasoconstriction and occasionally tongue vasoconstriction could be evoked by stimulating the internal carotid nerve. The postganglionic fibres innervating the nasal mucosa had a similar threshold to those of the nictitating membrane, which may indicate that there are small myelinated fibres innervating the mucosa.The preganglionic compound nerve action potential had four major components, S₁–S₄. S₁, S₂ and usually S₃ fibres were associated with membrane contraction; S₂, S₃ and sometimes S₁ fibres were associated with nasal vasoconstriction; and S₃, usually S₂ and occasionally S₁ fibres were associated with vasoconstriction in the tongue. It is concluded that each of these three groups of nerve fibres, but not S₄ fibres, may include fibres associated functionally with the three effectors.There was a considerable difference between the relative amplitude of the responses of the three effectors elicited by stimulation of the cervical sympathetic nerve at frequencies between 0.2 and 2 Hz. Vasoconstrictor responses were relatively larger than membrane contractions suggesting differences in the mechanisms of neurotransmission at the neuroeffector junctions.     

Development of the projection from the nucleus of the brachium of the inferior colliculus to the superior colliculus in the ferret

Neurons in the deeper layers of the superior colliculus (SC) have spatially tuned receptive fields that are arranged to form a map of auditory space. The spatial tuning of these neurons emerges gradually in an experience-dependent manner after the onset of hearing, but the relative contributions of peripheral and central factors in this process of maturation are unknown. We have studied the postnatal development of the projection to the ferret SC from the nucleus of the brachium of the inferior colliculus (nBIC), its main source of auditory input, to determine whether the emergence of auditory map topography can be attributed to anatomical rewiring of this projection. The pattern of retrograde labeling produced by injections of fluorescent microspheres in the SC on postnatal day (P) 0 and just after the age of hearing onset (P29), showed that the nBIC-SC projection is topographically organized in the rostrocaudal axis, along which sound azimuth is represented, from birth. Injections of biotinylated dextran amine-fluorescein into the nBIC at different ages (P30, 60, and 90) labeled axons with numerous terminals and en passant boutons throughout the deeper layers of the SC. This labeling covered the entire mediolateral extent of the SC, but, in keeping with the pattern of retrograde labeling following microsphere injections in the SC, was more restricted rostrocaudally. No systematic changes were observed with age. The stability of the nBIC-SC projection over this period suggests that developmental changes in auditory spatial tuning involve other processes, rather than a gross refinement of the projection from the nBIC.     

A contribution to the study of the architecture of the autonomic nervous system of the digestive tract of the rat

Summary The distribution of aminergic and non-aminergic nerve fibres to the different constituents of the wall of the digestive tract in various regions is described. Aminergic fibres synapse with all nervous perikarya. Densely interlacing networks of nerve fibres are found in both layers of the tunica muscularis and in the lamina muscularis mucosae. A finely meshed plexus is observed in relation to the wall of the blood vessels in the wall of the gut. There are many fibres connecting the muscular and the vascular plexus. No nerve fibres have been observed in direct relation to the epithelium.The functional implications of these findings are discussed.