半乳糖凝集素1在退变椎间盘中的差异表达

背景:半乳糖凝集素1调控诸多组织、器官的生理病理过程,其与椎间盘退变的关系尚不明确。目的:分析半乳糖凝集素1在不同退变程度椎间盘中的差异性表达,并探讨其对大鼠椎间盘退变的影响。方法:收集新疆医科大学第一附属医院15例患者的椎间盘标本,采用Pfirrmann分级后检测半乳糖凝集素1在不同退变程度椎间盘中的差异性表达。建立SD大鼠腰椎退变模型,随机分为3组,造模后隔日进行干预,对照组、模型组分别尾静脉注射生理盐水,半乳糖凝集素1干预组尾静脉注射半乳糖凝集素1重组蛋白,1次/d,各组均干预7 d;造模术后8周使用Bruker Pharmascan 7.0T核磁共振仪进行Pfirrmann分级后观察椎间盘的病理变化,并检测椎间盘中半乳糖凝集素1、Ⅱ型胶原及蛋白聚糖的表达。结果与结论:(1)qRT-PCR、免疫组织化学染色及Western Blot结果提示随着椎间盘退变的加重,半乳糖凝集素1表达逐渐减少,3组之间比较差异有显著性意义(P<0.05);(2)术后8周,大鼠腰椎MRI提示半乳糖凝集素1干预可降低改良Pfirrmann分级;苏木精-伊红染色提示半乳糖凝集素1干预减少椎间盘退变的病...     

双上肢去势联合椎间盘刺破诱导建立大鼠椎间盘退变模型

背景：寻求一种相对合适的椎间盘退变动物模型,对深入研究椎间盘退变的发病机制有着重要意义。目的：探索一种新的椎间盘退变动物模型,希望为椎间盘退变疾病的基础研究提供新的造模方式选择。方法：将20只雄性SD大鼠随机分为假手术组与模型组。采用双上肢去势联合椎间盘刺破的方式制备椎间盘退变模型,于造模当天及造模后4周、8周进行MRI检查,观察造模前后椎间盘形态变化并评估Pfirrmann分级;造模4,8周后取出大鼠L₄-L₅及L₅-L₆椎间盘,运用苏木精-伊红染色、Masson染色法观察椎间盘病理学变化及免疫组化观察Ⅱ型胶原蛋白表达量。结果与结论：(1)模型组在造模当天仅呈现轻微退变;(2)在造模第4周时与假手术组相比,模型组MRI上T2加权信号降低,在Pfirrmann分级上可见明显差异(P<0.05),在造模第8周时与假手术组相比,模型组MRI上T2加权信号明显降低,与纤维环边界模糊不清,且髓核向后纵韧带膨出,在Pfirrmann分级上可见显著差异(P<0.01);(3)病理染色结果显示,在造模...     

PINK1/Parkin介导的线粒体自噬在力学失衡诱导终板软骨退变中的作用

目的 检测脊柱失稳诱导终板软骨退变模型大鼠中线粒体自噬水平的变化,探讨PINK1/Parkin介导的线粒体自噬在终板软骨以及椎间盘退变中的作用。方法 通过手术切除大鼠L2～L5棘上、棘间韧带,咬除L2～L5两侧关节突,构建大鼠脊柱失稳模型。18只SD大鼠分为正常组、退变组及羰基氰化物3-氯苯腙（CCCP）组,每组6只。正常组大鼠无特殊处理,退变组大鼠构建大鼠脊柱失稳模型,CCCP组大鼠在构建大鼠脊柱失稳模型后于椎间盘注射5μL CCCP (10μmol/L)。利用HE染色观察终板软骨及椎间盘形态变化,番红-固绿染色观察终板软骨细胞外基质变化。RT-PCR检测各组大鼠终板软骨组织中Ⅱ型胶原（COL-2A）、蛋白聚糖（ACAN）、PINK1、Parkin mRNA表达水平;Western blot检测COL-2A、ACAN、PINK1、Parkin及线粒体膜蛋白Tomm20、Timm23蛋白表达水平变化。结果 与正常组比较,退变组大鼠椎间盘髓核破坏明显,终板软骨细胞外基质分泌减少;CCCP组大鼠椎间盘结构较完整,终板软骨细胞外基质分泌较退变组明显增多。与正常组比较,退变组大鼠终板软骨组织C...     

丁香苷抑制大鼠椎间盘退变

背景:椎间盘退变是由于椎间盘内部髓核和纤维环组织发生损伤和退化导致的椎间盘结构和功能发生变化,目前尚无有效的治疗药物。目的:探讨丁香苷抑制大鼠椎间盘退变的作用。方法:取10只雄性SD大鼠,将每只大鼠的尾椎Co_(4)/Co_(5)椎间盘设为模型组、Co_(5)/Co_(6)椎间盘设为丁香苷组、Co_(6)/Co_(7)椎间盘设为对照组,对照组不进行任何处理,模型组、丁香苷组采用微型穿刺针进行纤维环全层穿刺建立椎间盘退变模型,造模后即刻,模型组、丁香苷组椎间盘分别注射2.5μL的生理盐水、丁香苷溶液(5μmol/L)。注射4周后取材,采用苏木精-伊红和番红O-固绿染色观察大鼠椎间盘退变程度,免疫组化染色分析大鼠椎间盘组织内Ⅱ型胶原、聚集蛋白聚糖及基质金属蛋白酶3,13的表达。结果与结论:①苏木精-伊红染色显示,模型组椎间盘高度降低,软骨终板变薄且有裂隙出现,纤维环结构紊乱且出现裂隙,髓核消失;丁香苷组椎间盘高度正常或略低于对照组,软骨终板退变程度较模型组轻,纤维环排列较模型组相对规整且无裂隙,髓核部分皱缩。②番红O-固绿染色显示,模型组椎间盘软骨终板出现缺损且软骨钙化层变薄,出现明显退变;丁香苷组椎间盘软骨终板结构形态有一定程度恢复。③免疫组化染色显示,与对照组比较,模型组椎间盘软骨组织内Ⅱ型胶原、聚集蛋白聚糖的表达降低(P<0.0001),基质金属蛋白酶3,13的表达升高(P<0.0001);与模型组比较,丁香苷组椎间盘软骨组织内Ⅱ型胶原、聚集蛋白聚糖的表达升高(P<0.001,P<0.0001),基质金属蛋白酶3,13的表达降低(P<0.001,P<0.0001)。④结果表明,丁香苷可通过抑制基质金属蛋白酶3,13的表达、提高Ⅱ型胶原和聚集蛋白聚糖的表达来改善椎间盘的结构和功能,预防和减缓椎间盘退变过程。     

巴马小型猪胎儿成纤维细胞重编程为诱导多能性细胞的研究

目的探讨巴马小型猪胎儿成纤维细胞重编程为诱导多能性细胞的方法。方法应用经典的逆转录病毒介导方法,将Oct4,Sox2,c-Myc和Klf4四种人源性转录因子转导入巴马小型猪的胎儿成纤维细胞中,建立形态和特征类似猪上胚层细胞来源的猪多能性细胞(pig induced pluripotent cells,piPCs)。免疫荧光化学方法检测多能性细胞标记物,RT-PCR检测了该细胞体外三胚层相关基因表达水平。结果巴马小型猪胎儿成纤维细胞重编程的piPCs细胞呈AKP染色阳性,体外长期传代都保持正常的染色体核型,猪上胚层多潜能性细胞标记分子Oct4,Sox2,Nanog,Tra-1-60,Tra-1-81和SSEA-4呈阳性表达,并且表达小鼠胚胎干细胞(ES)多潜能性细胞标记分子SSEA-1,体外诱导分化后具有三胚层特异基因的表达,但是体内分化能力存在缺陷。结论巴马小型猪胎儿成纤维细胞重编程成功诱导成多能性细胞。     

转化生长因子β1注射退变椎间盘内软骨终板的组织形态变化

背景：椎间失稳能导致软骨终板的退变,是椎间盘退变发病机制中的基础环节。 目的：观察退变椎间盘内注射转化生长因子β1后,椎间盘软骨终板的组织形态学变化。 方法：将日本大耳白兔随机分为对照组、预防组和治疗组。所有兔均建立L5~6,L6~7椎间失稳模型。预防组在完成椎间失稳建模后立即于损伤侧L5~6,L6~7椎间盘内注射转化生长因子β1,治疗组于椎间失稳建模后3个月行相同方法注射转化生长因子β1。 结果与结论：建模后3,6个月,预防组较对照组软骨终板软骨细胞分布均匀,潮线清晰。Mankin评分降低(P < 0.05)。建模后第6个月治疗组较对照组软骨终板表层光滑,细胞排列均匀,潮线清晰,染色均匀,Mankin评分降低(P < 0.05)。结果证实,在兔椎间盘内注射转化生长因子β1可延缓椎间盘软骨终板的退变。     

间充质干细胞外泌体上调Mir-21调控HO-1机制促进椎间盘退变的修复

目的探讨间充质干细胞外泌体上调Mir-21调控血红素氧合酶(HO-1)机制促进椎间盘退变的修复研究。方法新西兰健康大白兔36只随机数字法分成3组,Blank组(正常大白兔)、Model组(椎间盘退变大白兔)和Mir-21组(椎间盘退变大白兔+间充质干细胞外泌体),通过建模处理、外泌体提存、免疫组织化学染色、Rt-PCR检测、HE染色、流式细胞术检测,观察外泌体形态、对比分析HO-1蛋白阳性率在椎间盘退变中的表达,Mir-21、HO-1和BACH1在大白兔椎间盘组织中的表达,大白兔椎间盘组织形态学变化,白兔退变髓核细胞凋亡能力情况。结果 Blank组HO-1蛋白阳性率为(62.23±4.1)%,细胞颗粒呈棕黄色,Model组的蛋白阳性率为(8.96±2.4)%,明显低于Blank组(P0.05),Mir-21组蛋白阳性率为(42.38±3.46)%,细胞颗粒呈淡黄色,显著高于Model组低于Blank组(P0.05);Model组的Mir-21、HO-1表达水平明显低于Blank组和Mir-21组(P0.05),Mir-21组的Mir-21、HO-1表达水平显著低于Blank组(P0.05)。Model组的BACH1水平明显高于Blank组和Mir-21组(P0.05),Mir-21组的BACH1水平显著高于Blank组(P0.05);Blank组兔髓核内基质比较高,髓核细胞颜色较深,周边胞浆颜色较淡,Model组通过建模后髓核细胞消失,与Blank组相比基质密度比较低,髓核细胞难见(P0.05),Mir-21组通过Mir-21转染后,与Model组相比髓核内基质明显升高,密度染色加深,髓核细胞出现,并且染色加深(P0.05);Model组和Mir-21组的细胞凋亡能力明显低于Blank组(P0.05),Mir-21组细胞凋亡能力显著高于Model组(P0.05)。结论间充质干细胞外泌体上调Mir-21,抑制BACH1来促进HO-1机制修复椎间盘退变的能力,对退变组织细胞具有促凋亡能力。     

骨形态发生蛋白2对退变椎间盘组织学和Ⅰ，Ⅱ型胶原的影响

背景：研究表明,退变椎间盘细胞外基质的主要变化是Ⅱ型胶原和蛋白多糖含量的减少和Ⅰ型胶原含量的增加。 目的：观察腺相关病毒介导的骨形态发生蛋白2基因(AAV-BMP-2)对兔退变腰椎间盘内髓核Ⅰ,Ⅱ型胶原的影响。 方法：将12只新西兰大白兔L2-3,L3-4,L4-5,L5-6椎间盘针刺制造退变模型后随机分为3组,每组4只。其中AAV-BMP-2组兔椎间盘注射AAV-BMP-2,AAV组兔椎间盘注射不携带目的基因的空载体,生理盐水组兔椎间盘注射生理盐水,注射后第8周处死动物取其相应的椎间盘常规石蜡包埋,组织切片,以苏木精-伊红染色观察其髓核组织学变化,免疫组织化学法检测髓核组织Ⅰ型、Ⅱ型胶原表达并进行半定量分析。 结果与结论：普通苏木精-伊红染色结果显示AAV-BMP-2组椎间盘内髓核细胞数目较多,呈单个或者簇状分布,髓核结构清晰,无纤维样组织填充。而AAV组和生理盐水组的组织结构相似,髓核内细胞数目少,髓核皱缩干瘪,细胞间为纤维样组织填充且排列紊乱。免疫组织化学染色显示：AAV-BMP-2组椎间盘髓核内Ⅱ型胶原的表达均高于AAV组和生理盐水组(P < 0.05);AAV-BMP-2组椎间盘髓核内Ⅰ型胶原的表达均低于AAV组和生理盐水组(P < 0.05)。结果说明,体内转染AAV-BMP-2能抑制椎间盘髓核细胞表达Ⅰ型胶原,促进椎间盘髓核细胞表达合成Ⅱ型胶原,提示维持椎间盘内胶原的含量、种类,可维持椎间盘内组织学的结构和形态,稳定髓核细胞生长环境,延缓椎间盘的退变。     

可控轴向压力致兔腰椎间盘退变模型的建立及评价

目的：建立可控轴向压力诱发的椎间盘退变动物模型并考察其特性。方法:取4月龄日本大白兔40只，将其随机分入5组（每组8只）。采用基于横穿椎体的克氏针的自制加压装置，分别对2-4组动物腰椎间盘施加10 kg压力1周、2周、4周及8周，第1组作为假手术对照组。Thompson分级法及兔腰椎间盘磁共振（MRI）评价退变程度，HE染色及Ⅰ、Ⅱ型胶原免疫组化染色考察其组织学改变。另外，构建纤维环损伤致兔腰椎间盘退变模型，并对标本行HE染色以供比较。结果:①Thompson分级：第2组7只均为Ⅱ级，第3组6只Ⅲ级，第4组5只为Ⅳ级，第5组4只动物退变程度达到Ⅴ级。②MRI评分显示，第2组椎间盘以轻度退变为主；第3组椎间盘主要为中度退变。第4组、第5组则以重度退变为主。③HE染色显示，该模型椎间盘各区域的退变情况较纤维环损伤模型更为均匀。④免疫组化染色显示，退变过程中椎间盘基质成分退化的趋势与人体相符。结论:可控轴向压力致兔腰椎间盘退变模型可以高效诱发椎间盘退变，尤其在轻、中度椎间盘退变方面具有优势。该模型可供深入研究，并用于椎间盘退变的治疗研究。     

Expression of TGF-β1 and Bax and their significance in intervertebral disc degeneration

目的 探讨退行性变椎间盘组织中TGF-β1和Bax的表达及其意义.方法 收集正常与退变椎间盘组织,并根据病理改变将退变椎间盘组织分成4级,采用HE、免疫组化、TUNEL染色和RT-PCR法进行研究.结果 免疫组化和RT-PCR均显示在正常组织中有TGF-β1表达,Bax只有微量表达;在病变组织中随病理分级加大TGF-β1随之增加,与正常组相比,差异有显著性;Bax也逐步增加,与正常组相比,差异有显著性,Bax表达升高与凋亡指数(AI)呈正相关性.结论 退变椎间盘的病理分级与TGF-β1的表达增高相关,由此调控了Bax的表达,导致了细胞凋亡,促进了椎间盘的退变.     

Topical organization of the projections of neurons of the substantia nigra and the ventral field of the tegmentum of the midbrain to the head of the caudate nucleus of the cat

Neuroscience and Behavioral Physiology -     

Asymmetry of the internal connections of the striate cortex of the cat in the projection zone of the center of the field of vision

Studies were carried out on the organization of the internal connections of the striate cortex in cats in the projection zone of the center (0–5°) of the field of vision by microintophoretic application of horseradish peroxidase to electrophysiologically identified orientational columns. The area containing neurons showing retrograde labeling in most cases extended in the mediolateral direction. Labeled cells were located in the upper (II, III) and lower (V, VI) layers of the cortex, and the shapes and orientations of the areas containing labeled neurons in these layers coincided. Spatial asymmetry was detected in the distribution of labeled neurons relative to the orientational column studied. Labeled cells were located predominantly medial to the columns, regardless of the distance from the projection of the area centralis. Considering the visuotopical map of field 17, the asymmetry detected here provides evidence that neurons in orientational columns have more extensive connections with neurons of the peripheral part of the cortex. An asymmetrical distribution of “silent” zones around the receptive fields of neurons in orientational columns is suggested, and that these appear to receive influences from the periphery of the visual field. Laboratory of Visual Physiology and Laboratory of Central Nervous System Morphology, I. P. Pavlov Institute of Physiology, Russian Academy of Sciences, 6 Makarov Bank, 199034 St. Petersburg, Russia. Translated from Fiziologicheskii Zhurnal imeni I. M. Sechenova, Vol. 82, No. 12, pp. 23–29, December, 1996.     

Characteristics of the topographic interrelation of the head of the pancreas and the duodenum and pathology of the organs of the pancreatoduodenal area

Seventy pancreatoduodenal complexes of 55 patients with chronic pancreatitis and tumours of this zone and 15 patients died from other diseases are studies histotopographically . The pieces of the pancreatic head tissue in the medial wall of the duodenum were found in 12 cases of the first group and in 4 control cases. The pancreatic tissue consisted either of all elements of this organ or cystically dilated ducts and seemed to infiltrate different layers of the duodenum wall. Three variants of the pancreatic head structure are suggested on the basis of anatomo-topographical interrelationships of the pancreatic head and duodenum. In 12 out of 14 cases chronic pancreatitis and carcinoma of organs of this zone were combined with the variants of the pancreatic head structure, in 2 cases there was a true heterotopy . Pathogenetic significance of these variants for the development of chronic pancreatitis is discussed.     

Influence of the level of the transmembrane potential on the dynamics of the extinction of the amplitude of the synaptic reactions of the snail under voltage clamping conditions

The influence of the level of the transmembrane potential on the dynamics of the extinction of the amplitudes of summary excitatory postsynaptic currents arising in identified giant parietal neurons in response to rhythmic stimulation of the intestinal nerve was investigated in a preparation of the isolated CNS of the common snail in order to identify the possibility of the participation of the postsynaptic element in synaptic plasticity. It was demonstrated that, at a greater value of the transmembrane potential, the decrease in the amplitudes of the postsynaptic currents which have been induced by rhythmic stimulation takes place more rapidly. It was also demonstrated that at a higher frequency of stimulation of the nerve, the effect of the influence of the membrane potential level on the dynamics of the synaptic reactions is more prominent. The data obtained may be regarded as an argument in favor of the possibility of the participation of the post-synapse in plasticity. This study was partially supported by the Russian Basic Research Fund (project No. 94-04-12209). Institute of Higher Nervous Activity and Neurophysiology, Russian Academy of Sciences, Moscow. Translated from Zhurnal Vysshei Nervnoi Deyatel'nosti imeni I. P. Pavlova, Vol. 45, No. 1, pp. 171–179, January–February, 1995.     

Effects of the lesion of the postcommissural part of the septum on the behavior of the rat.

The effects of the lesion of the postcommissural part of the septum on behavior of the rat has been studied. Results may be summarized as follows. An increase in the exploratory behavior in the open field which decreases rapidly; a decrease in the number of defecations in this test and a decrease in time leaving a dark environment for exploration. In the shuttle box test, no facilitation of the acquisition, but a permanent and quite significant increase in the intertrial activity has been found. We conclude that the lesions tend to decrease the emotivity of the subjects. An interpretation on the basis of the species -- specific defensive reactions explains the transitory and permanent effects of the lesions on the spontaneous activity.     

Effect of the conditions of adaptation of the optic analyzer to the function of the thermoreceptors of the human skin

Summary A discussion is presented of the effect produced by light stimulation applied to the optic analyzer alone on the function of the temperature analyzer. The cutaneous-temperature analyzer was kept on constant illumination and temperature. The light effects from the retinal photoreceptors on the cutaneous thermoceptor system led to reflex changes of the functional rate of the latter. Conditions of the optic analyzer of light adaptation caused reflex adjustment of cold receptors as to heating. Dark adaptation of the eyes changed the functional state of the cold receptors, corresponding to their adjustment to low temperatures.(Presented by Active Member AMN SSSR V. V. Parin) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 57, No. 3, pp. 3–6, March, 1964