电刺激小脑顶核促进大脑中动脉缺血鼠共移植体中神经干细胞向神经元的分化

目的:向大脑中动脉缺血大鼠植入神经干细胞,探讨电刺激小脑顶核与神经干细胞共移植体对其向神经元分化的影响。方法:实验于2005-08/2006-11在佳木斯大学神经科学研究所完成。①选取同一基因背景大鼠72只,随机数字表法分成3组:神经干细胞移植组、电刺激 神经干细胞组、电刺激 神经干细胞共移植体组,24只/组。②另选取出生24h内的Wistar鼠1只用于神经干细胞的分离,制备单细胞悬液,调整细胞密度为5×108L-1,置于10mg/L的Brdu完全培养液中孵育72h。神经干细胞共移植体由神经干细胞、层粘连蛋白和血管内皮细胞构成,由本实验室提供并鉴定。③各组大鼠于造模前1d麻醉后行小脑顶核刺激,以前囟为零点、正中线向后11.6mm、正中线向右1.2mm。给予直角方波脉冲刺激,电流强度为50μA,频率80Hz,时程1h。④各组大鼠均采用线栓法建立局灶性脑缺血动物模型。造模1d后,分别将培养的神经干细胞及共移植体细胞浓度调整为2×1010 L-1,于缺血纹状体侧缺血半暗带区垂直进针,进入5.0mm后缓慢推入细胞悬液,神经干细胞移植组、电刺激 神经干细胞组移植神经干细胞悬液10μL,电刺激 神经干细胞共移植体组移植神经干细胞共移植体悬液10μL,移植速度为0.5μL/min,留针10min后缓慢拔针。⑤各组分别于移植后3,7,14,60d观察神经干细胞移入脑内后分化成神经元的情况。胞核呈绿色荧光、胞质呈红色荧光的为Brdu-神经元特异性烯醇化酶双阳性细胞,代表移植入的神经干细胞所分化的神经元。结果:72只同一基因背景大鼠均进入结果分析。①与神经干细胞移植组比较,移植后7,14,60d电刺激 神经干细胞组、电刺激 神经干细胞共移植体组Brdu-神经元特异性烯醇化酶双阳性细胞百分率均明显升高(F=12.44～25.18,P均<0.05)。移植后3,7,14,60d电刺激 神经干细胞共移植体组Brdu-神经元特异性烯醇化酶双阳性细胞百分率均明显高于电刺激 神经干细胞组(F=8.19～25.18,P均<0.05)。②随移植时间的延长,各组Brdu-神经元特异性烯醇化酶双阳性细胞形态均逐渐增大,至移植后60d时细胞周边可见突起。结论:大脑中动脉缺血模型鼠移植入神经干细胞后,电刺激小脑顶核能够对神经干细胞向神经元的分化产生促进作用,且共移植体与电刺激小脑顶核具有协同效应。     

细胞移植治疗帕金森病的研究进展

目的:根据近年细胞移植治疗帕金森病的近况,认识细胞移植治疗帕金森病的最新进展及临床应用前必须解决的技术难题。资料来源:应用计算机检索Pubmed 2000-01/2006-10细胞移植治疗帕金森病的文章,检索词为"Cells transplantation,Parkinson disease",并限定文献种类为English。资料选择:对资料进行初审,选取包括细胞移植与帕金森病相关的文献,开始查找全文。纳入标准:①细胞移植在神经系统疾病中的应用。②细胞移植与帕金森病。排除标准:综述文献、重复研究、Meta分析类文章。资料提炼:共收集到175篇关于细胞移植与帕金森病的文献,纳入30篇文献。资料综合:细胞移植治疗帕金森病的主要细胞来源有肾上腺髓质细胞、胎儿中脑腹侧组织细胞、异种移植、神经干细胞、骨髓干细胞、胚胎干细胞等。其临床应用前仍存在许多技术难题,如移植物的来源、移植部位与方法、移植环境等。随着这些基础研究和临床治疗上难题的解决,细胞移植必将成为治疗帕金森病的重要手段。结论:细胞移植尤其是胚胎干细胞的应用,将为帕金森病的治疗带来新的曙光。     

优秀女子赛艇运动员比赛前后血乳酸、血尿素及血清肌酸激酶跟踪测试

目的:跟踪测试优秀女子赛艇运动员比赛前后血乳酸、血尿素及血清肌酸激酶的变化,探讨女子赛艇运动员机体能量物质代谢和赛后疲劳恢复的特点。方法:选取2005-07/09湖北省水上运动中心优秀女子运动员14名,其中国家健将级运动员7名,国家一级运动员7名;平均年龄(22±4)岁,训练年限(4.1±1.4)年,身高(174.2±3.1)cm,体质量(68.6±4.7)kg。14名运动员分别于2000m划艇比赛当日晨空腹、赛前15min、赛后5min,1h及24h抽取指尖血60μL,进行血乳酸、血尿素、血清肌酸激酶检测。结果:14名运动员全部进入结果分析。①比赛前后运动员血乳酸水平的变化:运动员空腹血乳酸水平为(1.22±0.45)mmol/L,赛前15min为(2.89±0.49)mmol/L。赛后5min血乳酸浓度高达(11.51±1.72)mmol/L,与空腹水平比较差异有显著性意义(t=3.077,P<0.01)。赛后24h血乳酸浓度显著下降至(1.76±0.24)mmol/L,与空腹水平基本接近(t=0.027,P>0.05)。②比赛前后运动员血尿素水平的变化:与空腹血尿素水平比较,运动员赛后5min血尿素浓度明显升高[(5.45±0.47),(6.13±1.00)mmol/L;t=2.416,P<0.05]。赛后24h血尿素浓度下降至(5.94±0.85)mmol/L,仍高于空腹水平(t=2.682,P<0.05)。③比赛前后运动员血清肌酸激酶活性的变化:与赛前比较,运动员赛后5min血清肌酸激酶活性明显升高[(3.38±1.58),(6.13±3.25)nkat/L;t=4.968,P<0.01]。赛后1h血清肌酸激酶活性开始下降,至赛后24h与赛前基本相似(t=1.537,P>0.05)。结论:①赛后5min血乳酸、血尿素、血清肌酸激酶活性显著高于赛前,赛后1h血乳酸消除迅速,但仍未恢复到正常水平。提示赛艇是一种以糖酵解系统为主、无氧 有氧代谢混合型供能的运动项目。②赛艇比赛使酸性产物生成增多,血乳酸、血尿素、血清肌酸激酶可作为运动强度和机能恢复的指标。③赛艇比赛后至少24h内,机体处于蛋白质降解增强状态,建议恢复期增加饮食糖和蛋白质摄入量,以促进合成代谢,加快功能恢复过程。     

白介素-8、白介素-15在结肠癌患者结肠黏膜的表达及意义

目的:研究结肠癌患者结肠黏膜中白介素-8(IL-8)、白介素-15(IL-15)的表达并探讨其临床应用价值．方法:采用免疫组化法检测结肠癌患者结肠黏膜中IL-8,IL-15的表达情况．结果:IL-8染色阳性44例,占66．7％．IL-15染色阳性40例,占60．6％．IL-8和IL-15表达水平与结肠癌临床分期(IL-8:r=0．437,P=0．006; IL-15:r=0．317,P=0．014)、浸润深度(IL-8: r=0．332,P=0．003;IL-15:r=0．312,P= 0．015)、淋巴结转移(IL-8:r=0．316,P=0．042; IL-15:r=0．236,P=0．017)、病理分级(IL-8:r =0．826,P=0．0001;IL-15:r=0．368,P=0．001)均呈显著正相关．结论:检测IL-8,IL-15的表达可做为判断结肠癌恶性程度有价值的指标．     

Mechanism for cotolerance in nonlethally conditioned mixed chimeras: negative selection of the Vbeta T-cell receptor repertoire by both host and donor bone marrow-derived cells

Bone marrow (BM) chimeras prepared by complete recipient ablation (A-- >B) exhibit donor-specific tolerance, yet survival is often limited by graft-versus-host disease (GVHD). Negative selection of potentially donor-reactive T cells, as assessed by relative T-cell receptor (TCR)- Vbeta expression, is dependent on donor BM-derived deleting ligands. Mixed chimerism and tolerance for both donor and host antigens can be achieved using partial recipient myeloablation with 500 cGy total-body irradiation (TBI) before transplantation followed by cyclophosphamide (CyP) on day +2. To examine the influence of residual host elements on negative selection, the peripheral TCR-Vbeta repertoire was analyzed in partially ablated C57BL/10SnJ (B10) recipients reconstituted with BM from major histocompatibility complex (MHC)-disparate B10.BR/SgSnJ or MHC, Hh-1 and Mls-disparate BALB/cByJ donors, which delete Vbeta5+ and 11+ or Vbeta3+, 5+, and 11+ TCR subsets, respectively. As in myeloblated recipients, donor-reactive subfamilies were deleted in B10.BR-->B10 and BALB/c-->B10 chimeras, suggesting that donor I-E and minor lymphocyte-stimulating (Mls) antigens contribute to the deleting ligands in the nonmyeloablated host. In striking contrast to completely ablated B10-->B10.BR chimeras, partially ablated recipients showed intramedullary I-E expression in the thymus and deleted host-reactive Vbeta5+ and Vbeta11+ subfamilies. These data demonstrate that efficient clonal deletion occurs after partial myeloablation and that both donor and host ligands contribute to TCR repertoire selection.     

Protective effect of YHI and HHII against experimental acute pancreatitis in rabbits

AIM: To observe the protective effect of combined i.v. administraction of Yuanhu injection (YHI) and Huoxuehuayu injection-I (HHI-I) against acute pancreatitis (AP) in rabbits.METHODS: Sever acute pancreatitis (SAP) was induced by retrograde infusion of artificial bile juice into biliary-pancreatic duct, and treated with YHI and HHI-I intravenously. The protective effect was judged by the survival time and rate, serum amylase, serum interleukin-6, pancreatic microcirculation and pathological alteration.RESULTS: Combined use of YHI and HHI-I could markedly increase the rabbits’ 5-d survival rate after AP (83.3% in the treatment group and 33.3% in control). The serum amylase value (x^- ± s) decreased to 1596.6 U/L ± 760.50 U/L in the 5th day from the high level (6320.83 U/L ± 2614.12 U/L) in the 1st day after AP in the treatment group, while in the control group the amylase activity in the 5th day was 2095.0 U/L ± 1081.87 U/L, being significantly different from that before AP (837.17 U/L ± 189.12 U/L). YHI and HHI-I also obviously improved the pancreatic microcirculation and lowered the serum interleukin-6 level, one of the indices of severe pancreatitis. Pathological examination indicated all the changes typical for AP in YHI and HHI-I treatment group were milder than those in the control.CONCLUSION: YHI and HHI-I used in combination might have protective effect against acute pancreatitis in rabbits.     

Hypervitaminosis A-induced liver fibrosis: stellate cell activation and daily dose consumption.

Hypervitaminosis A-related liver toxicity may be severe and may even lead to cirrhosis. In the normal liver, vitamin A is stored in hepatic stellate cells (HSC), which are prone to becoming activated and acquiring a myofibroblast-like phenotype, producing large amounts of extracellular matrix. AIMS: In order to assess the relationship between vitamin A intake, HSC activation and fibrosis, we studied nine liver biopsies from patients belonging to a well-characterized series of 41 patients with vitamin A hepatotoxicity. METHODS: Fibrosis was underlined by Sirius-red staining, whereas activated HSC were immunohistochemically identified using an antibody against alpha smooth muscle actin. The volume density (Vv) of sinusoidal and total fibrosis and of sinusoidal and total activated HSC was quantified by the point-counting method. RESULTS: Morphology ranged from HSC hypertrophy and hyperplasia as the sole features to severe architectural distortion. There was a significant positive correlation between Vv of perisinusoidal fibrosis and the daily consumption of vitamin A (P=0.004). CONCLUSION: The close correlation between the severity of perisinusoidal fibrosis and the daily dose of the retinol intake suggests the existence of a dose-effect relationship.     

Heterogeneity of neutrophil antibodies in patients with primary Sjogren's syndrome

Our aims were to determine the prevalence of neutrophil antibodies in patients with primary Sjogren's syndrome (pSS), identify their target antigen(s), and evaluate their functional significance. Neutrophil antibodies were detected using an indirect immunofluorescence (IIIF) test and an enzyme-linked immunosorbent assay (ELISA), using recombinant human Fc-gamma receptor (Fc gamma RIIIb) as a capture agent. Luminol-dependent chemiluminescence was then measured by an established technique. Antibodies to neutrophils were detected in 30 of 66 patients (45%) and categorized on the basis of positivity for the two assays: IIF+/ELISA+ (group A: five patients), IIF+/ELISA- (group B: five patients), and IFF-/ELISA+ (group C: 20 patients). All positive sera contained antibodies directed to the neutrophil specific Fc gamma RIIIb, and none of them bound to NAnull neutrophils. The titer of neutrophil-reactive antibodies (groups A and B) showed no correlation with the neutrophil count, but these autoantibodies did reduce the cell ability to generate a respiratory burst. Thus, neutrophil antibodies are common in patients with pSS. Their main target appears to be Fc gamma RIII, and this may partly account for the dysfunction in Fc gamma R-mediated clearance by the reticuloendothelial system reported in these patients.     

Long-term favourable outcome of portal hypertension complicating primary systemic amyloidosis

Abstract: An unusual case of systemic amyloidosis complicated by portal hypertension is reported. In this condition, portal hypertension is a rare event associated with poor prognosis. In our patient, severe presymptomatic sinusoidal portal hypertension was demonstrated by hepatic vein catheterization and coincided with abundant perisinusoidal amyloid infiltration. Despite these features and the absence of objective response to 20 courses of melphalan and prednisone, the patient was still in good clinical condition 4 years after initial diagnosis. This observation suggests that in primary amyloidosis, the incidence of sinusoidal portal hypertension might be underestimated and also that it may be associated with a relatively good prognosis.