自组装肽包裹嗅神经鞘细胞抑制脊髓损伤后胶质瘢痕形成并促进轴突再生

目的研究自组装肽包裹嗅神经鞘细胞(OECs)移植对大鼠脊髓损伤(SCI)后轴突再生的影响。方法成年雄性SD大鼠40只,行脊髓T_(10)节段背侧横断术,动物随机分为4组,每组10只。SAP-OECs组于间隙内移植SAP包裹的OECs悬液,SAp、OECs对照组同法分别移植等量SAP及DF12-OECs悬液;单纯全切对照组不做处理。术后2或4周过量麻醉处死动物,脊髓损伤节段连续冰冻矢状切片。行抗GAP-43和GFAP免疫组织化学和免疫荧光双标染色。结果SAP-0ECs组损伤区内有大量GAP-43免疫反应纤维,SAP组相较OECs及单纯半切对照组则可见相当数量GAP-43免疫反应纤维。OECs及单纯半切对照组损伤区周围可见一条致密度的GFAP阳性的反应性星形胶质细胞条带,而SAP组GFAP免疫反应纤维比对照组少,且多于SAP-OECs组。此外激光共聚焦显微镜下观察SAP-OECs组在损伤后4周GAP-43免疫反应纤维能通过位于脊髓损伤区尾端的GFAP阳性区域。结论自组装肽包裹嗅鞘细胞可以抑制脊髓损伤后胶质疤痕形成并促进轴突再生。     

移植微囊化嗅鞘细胞对P2X2、P2X3受体介导神经病理性疼痛影响的实验研究

目的 探讨微囊化嗅鞘细胞移植对大鼠P2X2、P2X3受体介导神经病理性疼痛在L_4～5段脊髓表达的影响。方法 2014年9月—2015年7月采用差速贴壁法培养SD大鼠嗅球来源的嗅鞘细胞并进行传代扩增,取第9代嗅鞘细胞免疫荧光染色鉴定细胞纯度,符合要求后制作微囊化嗅鞘细胞。按随机化要求将48只健康SD大鼠随机分成对照组、模型组、嗅鞘细胞移植组(OECs组)、微囊化嗅鞘细胞移植组(MC-OECs组),每组12只。模型组、OECs组、MC-OECs组建立SD大鼠坐骨神经损伤的动物模型,OECs组、MC-OECs组分别将吸附嗅鞘细胞悬液和微囊化嗅鞘细胞悬液的可吸收明胶海绵置于坐骨神经损伤两侧,于术后第7、14天取各组SD大鼠的L_4～5段脊髓,采用原位杂交技术观察对比各组P2X2、P2X3受体阳性细胞的表达情况。结果 原位杂交结果显示,P2X2、P2X3受体主要表达于脊髓灰质中,以细胞核表达明显,细胞质内少量表达。术后第7、14天模型组、OECs组、MC-OECs组和对照组P2X2、P2X3受体的阳性细胞百分比及细胞平均吸光度均依次降低,且各组间比较,差异均有统计学意义(P值均<0.05)。结论 MC-OECs移植能明显降低坐骨神经损伤动物模型脊髓内P2X2、P2X3受体的表达水平,具有抑制P2X2、P2X3受体介导的神经病理性疼痛的作用。     

OECs移植联合IN-1抗体修复急性脊髓损伤的实验研究

目的研究嗅鞘细胞移植联合轴突生长抑制蛋白抗体IN-1局部持续注射对大鼠横断性脊髓损伤的修复作用.方法成年雄性SD大鼠40只,建立胸脊髓全横断损伤模型,随机分成单纯对照组(10只)、嗅鞘细胞移植组(10只)、IN-1抗体微泵注射组(10只)和嗅鞘细胞移植联合IN-1抗体组(10只).应用NF200免疫组化染色和免疫荧光染色对脊髓损伤区神经纤维再生进行形态学观察.采用BBB评分评估大鼠后肢功能恢复情况.结果横断损伤共有9只大鼠死亡.术后8周可观察到Hoeehet标记的嗅鞘细胞在体内存活并在脊髓内迁移;联合治疗(OECs IN-1)组和OECs组可见脊髓损伤区杂乱无序的再生轴突,但无连续性神经纤维通过损伤区;IN-1组和对照组脊髓残端萎缩,未见轴突再生.后肢功能运动平均BBB评分对照组、IN-1抗体组、细胞移植组和联合治疗组分别为7.70±0.24、7.89±0.15、10.50±0.25、11.33±0.24.结论OECs移植联合IN-1抗体可促进损伤的脊髓神经轴突的存活和再生,较单纯应用OECs或IN-1能更好的促进脊髓损伤修复和大鼠后肢功能恢复.     

纤维蛋白胶包裹嗅神经鞘细胞促进脊髓轴突再生的实验研究

目的 观察纤维蛋白胶(FG)包裹嗅神经鞘细胞(OECs)植于成年大鼠脊髓全横断断端间隙内对损伤轴突再生的促进作用。方法 成年雄性SD大鼠16只，行脊髓T11节段全横断术。FG-OECs组于间隙内移植FG包裹的OECs，FG对照组移植等量的FG，OECs对照组移植等量DF12-OECs悬液，单纯全切对照组未做任何移植，每组动物4只。术后2周处死动物，脊髓损伤区冰冻水平切片，荧光显微镜下观察OECs的存活及迁移，并行抗神经丝(NF)和生长相关蛋白-43(GAP-43)抗体免疫组织化学染色。结果 OECs及单纯全切对照组脊髓两断端连接较差，而FG对照组及FG-OECs组两断端连接良好。FG、OECs及单纯全切对照组损伤区可见少量NF及GAP-43免疫反应纤维；FG-OECs组间隙内OECs大量存活，并迁移至两侧脊髓实质内，大量NF及GAP-43免疫反应纤维通过间隙。结论 FG包裹的OECs可在成年大鼠脊髓全横断断端间隙内存活并迁移至脊髓实质内，促进损伤轴突的再生。     

人骨髓间质干细胞和嗅鞘细胞移植对大鼠急性脊髓损伤功能修复的比较研究

目的对比人骨髓基质细胞(BMSCs)与人胚嗅鞘细胞(OECs)移植对大鼠脊髓损伤功能修复的影响。方法将45只SD大鼠分成脊髓损伤后BMSCs移植组(BMSCs组)、OECs移植组(OECs组)和PBS对照组(PBS组),通过BBB评分、运动诱发电位(MEP)评估脊髓传导功能的改善状况,免疫组织化学方法检测移植细胞存活和分化情况,病理形态学方法观察组织结构修复情况。结果BMSCs组BBB评分高于OECs组(P<0.05);两细胞治疗组MEP潜伏期明显缩短(P<0.05);BMSCs组嗜银染色可见脊髓损伤近端有较多再生纤维向远端延伸,形成神经纤维束,而OECs组再生纤维较少;两种移植细胞均可在损伤处部分存活,BMSCs组可见BMSCs来源的细胞Nestin、NF、GFAP的阳性表达。结论BMSCs移植比OECs移植能更有效促进大鼠急性脊髓损伤修复。     

嗅鞘细胞移植和左旋硝基精氨酸对大鼠脊髓损伤后背核神经元存活及其轴突再生的影响

目的 探讨嗅鞘细胞(OECs)移植和一氧化氮合酶(NOS)抑制剂左旋硝基精氨酸(LNNA)对大鼠脊髓损伤后,背核神经元存活及其轴突再生的影响.方法 将20只大鼠行脊髓半横断术后分为对照组、OECs组、L-NNA组和OECs L-NNA组.OECs L-NNA组在脊髓损伤处移植嗅鞘细胞,并注射L-NNA.术后30d时,应用NADPH酶组织化学、免疫组织化学、中性红染色、HE染色和荧光金逆行标记等方法,观察4组大鼠L1背核神经元存活及其轴突再生情况.结果 1.0ECs组、L-NNA组L1背核神经元存活数量均高于对照组,且分类计数显示,这两组对脊髓背核大、中、小神经元的存活均有促进作用,而OECs L-NNA组的促存活作用最显著.2.0ECs组和OECs L-NNA组脊髓损伤处可见再生的轴突,且L1背核可观察到被荧光金逆行标记的神经元胞体.3.与对照组比较,OECs组L1背核NOS阳性神经元增加,L-NNA组NOS阳性神经元减少,OECs L-NNA组NOS阳性神经元数量差异不明显.结论 嗅鞘细胞移植和L-NNA均可促进脊髓受损伤背核神经元的存活,两者联合应用可更好地促进受损伤的背核神经元存活及其轴突再生.     

嗅成鞘细胞移植与甲基强的松龙联合应用对急性脊髓损伤大鼠功能恢复和Nogo-A表达的影响

目的:探讨嗅成鞘细胞(OECs)移植联合应用甲基强的松龙(MP)对急性脊髓损伤大鼠神经功能恢复及Nogo-A表达的影响.方法:自新生SD大鼠嗅球取材,纯化培养嗅成鞘细胞;建立大鼠脊髓挫伤模型,术后1、 3、 7、 14d,参照BBB法进行各组神经功能检测,取脊髓组织,免疫组织化学及RT-PCR法检测Nogo-A表达.结果:OECs组和MP组在7、14d BBB评分高于SCI组,但低于OECs+MP组;免疫组织化学和RT-PCR显示:在3、7、14d,OECs组和MP组Nogo-A的表达量与SCI组相比,均呈明显下调;但其下调Nogo-A效应低于OECs+MP组.结论:OECs移植联合应用MP可明显改善大鼠急性脊髓损伤后神经功能恢复,其机制部分与下调Nogo-A表达有关;OECs移植与MP联合应用在促进SCI后神经功能恢复方面要优于单独使用OECs或MP,两者之间可能存在协同效应.     

成年大鼠嗅球和鼻腔嗅粘膜成鞘细胞的分离、培养与鉴定

目的　在嗅球成鞘细胞 (OECs)移植到脊髓可有助于损伤神经纤维再生的基础上 ,分别对嗅球和嗅粘膜的OECs进行培养 ,探索自体嗅粘膜作为OECs供体的可能性。　方法　根据OECs、成纤维细胞和星形胶质细胞贴壁时间的不同 ,采用差时贴壁方法分离出OECs,培养 14div后进行NGFRp75和GDNF的免疫细胞化学染色。　结果　按形态学和免疫组织化学特性 ,培养的嗅球和嗅粘膜OECs可分为 3类 :双极细胞、三级细胞和扁圆细胞 ,其中以双极细胞最多。嗅粘膜的双极成鞘细胞的突起更加细长。　结论　差时贴壁细胞分离法是一种简单、经济、实用的成鞘细胞分离方法。鼻腔嗅粘膜OECs的形态学和免疫细胞化学特性与嗅球OECs基本相同 ,本实验为临床开展自体嗅粘膜OECs修复脊髓损伤的研究提供参考     

重组睫状神经营养因子对受损周围神经施万细胞相关基因表达的作用

目的研究重组睫状神经营养因子(CNTF)对受损周围神经施万细胞基因表达的作用。方法用硅管套接切断的大鼠坐骨神经，在受损神经局部给予重组CNTF，术后用免疫组织化学ABC法结合计算机图像分析观测S100蛋白(S100)、生长相关蛋白-43(GAP-43)、磷酸化酪氨酸(PTyr)、信号转导子和转录激活子(STAT)3的免疫反应阳性物质在修复侧远段神经的分布和相对含量。结果CNTF组修复侧远段神经相应区域S100、GAP-43、PTyr、STAT3阳性物质的含量显著或非常显著高于生理盐水组。结论重组CNTF能上调受损神经施万细胞S100、GAP-43、PTyr和STAT3的表达，提示重组CNTF通过强化受损神经施万细胞JAK-STAT途径，E调其S100和GAP-43的基因表达。     

移植微囊化嗅鞘细胞对神经病理性疼痛及L4-5段脊髓P2X7受体表达的影响

目的 探讨微囊化嗅鞘细胞移植对坐骨神经损伤引起病理性疼痛及L4-5段脊髓P2X7受体的表达影响。 方法 取1只健康SD大鼠,取其嗅球组织并运用差速贴壁方法,在体外进行嗅鞘细胞培养扩增。取48只健康SD大鼠,随机分成4组,即对照组（control）、慢性压迫性坐骨神经损伤组（CCI）、嗅鞘细胞移植组（OECs）及微囊化嗅鞘细胞移植组（MC-OECs）。术后7 d及14 d,运用行为学方法检测各组大鼠机械缩足反射阈值,运用原位杂交方法检测各组大鼠L4-5段脊髓P2X7受体阳性细胞百分比及平均吸光度表达情况。 结果 术后7 d及14 d,与对照组比较,CCI组大鼠机械缩足反射阈值明显减低,L4-5段脊髓P2X7受体阳性细胞百分比及平均吸光度明显增加（P<0.05）;与CCI组比,OECs组大鼠机械缩足反射阈值明显增高,P2X7受体阳性细胞百分比及平均吸光度明显减低（P<0.05）,与OECs组对比,MC-OECs组大鼠机械缩足反射更高。P2X7受体阳性细胞百分比及平均吸光度更低（P<0.05）。 结论 微囊化嗅鞘细胞移植能更好地缓解神经病理性疼痛,减低L4-5段脊髓内P2X7受体的表达水平,修复坐骨神经损伤。     

Apolipoprotein(a) isoform-specific changes of lipoprotein(a) after kidney transplantation

The atherogenic lipoprotein(a) (Lp(a)) is significantly increased in patients with kidney disease. Some studies in hemodialysis patients described this increase to be dependent on the genetic apolipoprotein(a) (apo(a)) isoforms. Only patients who express high molecular weight (HMW) apo(a) isoforms but not those with low molecular weight (LMW) isoforms show a relative increase of Lp(a) when compared to healthy controls matched for apo(a) isoforms. However, this was not confirmed by all studies. We therefore prospectively investigated the changes of Lp(a) deriving from each apo(a) isoform in heterozygotes following kidney transplantation. Lp(a) concentrations were measured by ELISA. To calculate the isoform-specific concentrations and the changes of Lp(a) deriving from each isoform, we densitometrically scanned the apo(a) bands from immunoblots before and after transplantation in 20 patients expressing two apo(a) isoforms. Of these, 10 patients expressed both an LMW and an HMW apo(a) isoform. The other 10 patients expressed only HMW isoforms. Densitometric scanning of apo(a) bands and calculation of isoform-derived Lp(a) concentrations clearly demonstrated that the decrease of Lp(a) following kidney transplantation is caused by changes in the expression of HMW apo(a) isoforms. In some patients, we observed an almost complete disappearance of the HMW apo(a) isoform after transplantation. This study clearly demonstrates that the changes of Lp(a) plasma concentrations in kidney disease depend on the genetically determined size of apo(a). This provides evidence for an interaction of apo(a) genetic variability and kidney function on Lp(a) concentrations.     

Lung alterations after paraquat poisoning (author's transl)

After one application of paraquat (35 mg/kg body weight) studies of the changes in lung structure in phospholipid content of the alveolar wash as well as in the static pV-curves are performed on adult hooded rats. Pathomorphologically 2, 4, 8, 16, 24 and 48 h after the paraquat intoxication focal atelectases, circumscribed inflammatory infiltrations of the interstitium are predominant as well as an accumulation of macrophages in the interstitium and in the alveolar space. Initially granulocytes dominate in the inflammatory infiltrates; after 8 h lymphoid elements and histiocytes became more and more apparent. 24 an 48 h after the paraquat injection there is a degradation of the alveolar epithelium. After 8-48 h the phospholipid content of the alveolar wash and after 2-48 h the hysteresis loop of the pV-curves are statistically significantly decreased in comparison with control animals. It is assumed, that the paraquat mediated dearrangement of the surfactant system of the lung is caused by an increased degradation in consequence of liberated phospholipases as well as by a decreased or ceased synthesis of the surfactant. The latter may be the consequence of an autooxydative membrane alteration of the endoplasmatic reticulum, the toxic action of the hydrogen peroxide originated by the autooxydation but also of the loss of energy rich substrates.     

心脏手术后精神障碍的临床心理学研究

近三十年来的研究表明 ,心脏手术后 ,尤其是体外循环下的心脏手术后精神障碍发病率非常高 ,症状容易被手术创伤所掩盖 ,易被临床所忽视 ,常常导致延误诊断或漏诊 ;其临床表现可在术后即刻出现或延迟数小时才出现 ,症状可以很轻微 ,在短时间内即可恢复 ,也可以很严重 ,导致病人死亡或造成病残 ,病程长的可持续数月至数年[1 ,2 ] 。现有的国内研究主要集中于其病因、发病率、一般性的预防与治疗上[3,4 ] ,很少在临床心理学方面进行深入研究 ,随机和前瞻性研究较少 ,尤其是对精神障碍的正确诊断和神经心理评估在预测脑损伤的作用上还认识不足 ,…     

Exophiala (Wangiella) dermatitidis keratitis after keratoplasty.

We report the first French case of an intraocular infection due to Exophiala (Wangiella) dermatitidis. Two months after a second corneal transplant for congenital hereditary endothelial dystrophy, the patient presented with ocular pain and corneal infiltrates leading to the graft rejection. Diagnosis was established by positive direct examination and cultures of the same fungus from corneal buttons, iris biopsies and ablated sutures.     

Rheumatoid course of humoral (vascular) rejection after heart allotransplantation

Analysis of planned endomyocardial biopsy specimens of heart allotransplants from 22 recipients revealed signs of humoral type rejection (slight, medium, and severe) presenting as fixation of IgG, IgM, and complement components (C3, C4d) in 61 of 63 sections. Permanent presence of rejection signs attests to rheumatoid course of this process. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 142, No. 9, pp. 337–340, September, 2006     

Bioencapsulation of living bacteria (Escherichia coli) with poly(silicate) after transformation with silicatein-alpha gene

Bioencapsulation is an intriguing way to immobilize biological materials, including cells, in silica, metal-oxides or hybrid sol-gel polymers. Until now only the sol-gel precursor technology was utilized to immobilize bacteria or yeast cells in silica. With the discovery of silicatein, an enzyme from demosponges that catalyzes the formation of poly(silicate), it became possible to synthesize poly(silicate) under physiological (ambient) conditions. Here we show that Escherichia coli can be transformed with the silicatein gene, its expression level in the presence of isopropyl beta-D-thiogalactopyranoside (IPTG) can be efficiently intensified by co-incubation with silicic acid. This effect could be demonstrated on the level of recombinant protein synthesis as well as by immunostaining analysis. The heterologously produced silicatein is enzymatically active, as confirmed by staining with Rhodamine 123 (formation for poly[silicate] from silicic acid) and by reacting free silicic acid with the beta-silicomolybdato color system. Electron microscopic analysis revealed that the bacteria that express silicatein form a viscous cover around them when growing in the presence of silicic acid. Finally, we demonstrate that the growth kinetics of E. coli remains unaffected whether or not the bacteria had been transformed with silicatein or grown in medium, supplemented with silicic acid. It is concluded that silicatein-mediated encapsulation of bacteria with silica might improve, extend and optimize the range of application of bacteria for the production of recombinant protein.