NOD/SCID孕鼠淋巴细胞表型检测和NK细胞活性分析

目的观察NOD/SCID小鼠免疫状况和生育力特征,并分析NK细胞亚群对同基因交配组合NOD/SCID×NOD/SCID妊娠结局的影响。方法采用流式细胞术对未孕和孕13.5 d的NOD/SCID小鼠进行淋巴细胞表型分析,并系统地观察和比较NOD/SCID×NOD/SCID和BALB/c×BALB/c小鼠的生育力特征。分别采用多聚次黄苷酸-胞苷酸(polyinosinic-polycytidylic acid,poly I∶C)或抗asialoGM1单抗(anti-asialo GM1)刺激或抑制NK细胞活性,并分别观察这些因素对妊娠结局的影响。结果与对照组NOD/SCID小鼠相比,poly I∶C处理组NOD/SCID×NOD/SCID小鼠平均每窝产仔数增多,而注射抗asialo GM1单抗则使胚胎吸收率增高,进而平均每窝产仔数显著减少。结论在NOD/SCID小鼠孕期母-胎界面保持适当水平的NK细胞活性对妊娠结局有利。     

地塞米松对谷氨酸钠致痫大鼠脑电图及GABA免疫反应的影响

为了探讨糖皮质激素的抑痫效应 ,本研究用免疫细胞化学方法和脑电图仪观察了地塞米松对谷氨酸钠致痫大鼠脑电图以及 GABA免疫反应的影响。结果显示 :大鼠在注射谷氨酸钠前 3 0 min经腹腔注射地塞米松 ,能明显减轻严重的癫痫发作症状和脑电图中高电位的痫波发放 ,并使海马齿状回及大脑皮层 GABA阳性神经元表达明显增强。显微图象统计分析 :与致痫组相比 ,抑痫组 GABA阳性神经元平均光密度值增加 ,差异有显著性 (P<0 .0 5 )。上述结果提示 :地塞米松具有抑制谷氨酸钠诱发癫痫的作用 ,其抑痫效应可能与增强 GABA的表达有关。     

肝癌组织差异表达基因cDNA序列的筛选与鉴定

目的：筛选并鉴定肝癌组织特异表达基因。方法：通过菌落原位杂交技术筛选用抑制消减杂交法构建肝癌与癌旁肝组织差异表达基因消减cDNA文库，用PCR方法进一步筛选出有插入片段的阳性克隆，将阳性克隆进行DNA测序和同源性比较分析，用Northern印迹方法对新的cDNA序列进行初步鉴定。结果：从消减文库中随机挑取的100个白色克隆中筛选出13个阳性克隆，DNA测序获得11个不同的cDNA序列；同源性比较分析表明，6个cDNA片段与在基因高度同源，5个cDNA片段为新的序列。其长度大于300bp的3个新序列，Norther印迹证实它都来源于肝癌组织。结论：用抑制消减杂交方法构建的肝癌差异表达基因消减cDNA文库富含肝癌特异表达基因，经验证的3个新的cDNA序列可能为肝癌特异的基因序列。     

浙南地区无偿献血者戊型肝炎亚临床感染及病毒血症情况调查

目的 调查浙江南部地区戊型肝炎的感染情况.方法 对2008年2-10月间浙江省温州市中心血站的无偿献血者进行整群抽样,检测抗HEV IgG以及IgM抗体,并对抗HEV IgM阳性的标本进行RT-PCR检测,经测序后分析其基因型及序列同源性.结果 3044例无偿献血者中,HEVIsG阳性率为33.28%,随着年龄的升高IgG阳性率也有一定的提高,从20岁组的21.16%升高到40岁以上组的50.36%;IgM阳性率0.92%,以31～40岁年龄段感染率最高,为1.90%.通过对IgG和lgM各自阴阳性标本群的分析发现其与献血者的年龄、性别以及血型并无显著相关性.对28例HEV IgM阳性标本进行PCR检测共发现3例核酸阳性,总体阳性率约为10%,其中2例为基因4型,1例为基因1型感染.结论 浙南地区的无偿献血者中存在一定比率的HEV病毒携带者.     

Deltorphin1单克隆抗体的制备及Deltorphin1样免疫活性在在体和离体大鼠脑神经元的表达

本研究利用结合到 BSA上的 deltorphin 肽 (DADT )免疫 Balb/c小鼠 ,成功地制备了特异性抗 DADT 单克隆抗体。交叉试验表明 ,所获抗体仅识别 DADT 和 DADT 而不识别其 L型同分异构体 L ADT ,也与其它 DADT 类似物无交叉反应。Epitope分析表明 ,该抗体识别 DADT 的 N末端特异氨基酸序列 ,其中包括对其活性有重要影响的第二位的 D-Ala-nine。利用这一抗体对成年大鼠脑组织和新生大鼠培养神经元进行了免疫组织化学研究。大鼠脑中有含量较低的 DADT 样免疫活性 ,主要分布在某些特定部位的神经纤维和神经元胞体 ,如终纹床核、杏仁核、海马、伏核、下丘脑外侧区、黑质、中央灰质等部位。然而在培养的大鼠脑神经元 ,DADT 样免疫活性的含量却很丰富 ,神经元的胞体和突起均呈较强的 DADT 样免疫活性。两项结果均提示哺乳动物脑神经元很可能产生一种与 DADT 在结构上极其类似的物质     

不同表面处理方法对纯钛表面形貌及成分的影响

分别用HNO3、热H2SO4/H2O2、热H2SO4/HC l处理纯钛片30 m in。采用扫描电子显微镜、X射线光电子能谱对试样的表面形貌及成分进行分析。扫描电镜结果显示,HNO3组表面形貌光滑,平整;而H2SO4/HC l、H2SO4/H2O2处理方法可获得粗糙的表面,其中H2SO4/HC l处理后的表面孔隙更大。X射线光电子能谱分析表明:三组钛片表面的主要成分均为钛、氧和碳,H2SO4/H2O2组的碳含量最低,而H2SO4/HC l组碳含量最高。HNO3组和H2SO4/H2O2组表面除了T iO2还存在T i2O3、T iO、金属T i等多种物质,而H2SO4/HC l组表面只存在T iO2。     

Loss of leucine-rich repeat kinase 2 causes impairment of protein degradation pathways,accumulation of α-synuclein,and apoptotic cell death in aged mice

Mutations in leucine-rich repeat kinase 2 (LRRK2) are the most common genetic cause of Parkinson''s disease. LRRK2 is a large protein containing a small GTPase domain and a kinase domain, but its physiological role is unknown. To identify the normal function of LRRK2 in vivo, we generated two independent lines of germ-line deletion mice. The dopaminergic system of LRRK2^−/− mice appears normal, and numbers of dopaminergic neurons and levels of striatal dopamine are unchanged. However, LRRK2^−/− kidneys, which suffer the greatest loss of LRRK compared with other organs, develop striking accumulation and aggregation of α-synuclein and ubiquitinated proteins at 20 months of age. The autophagy–lysosomal pathway is also impaired in the absence of LRRK2, as indicated by accumulation of lipofuscin granules as well as altered levels of LC3-II and p62. Furthermore, loss of LRRK2 dramatically increases apoptotic cell death, inflammatory responses, and oxidative damage. Collectively, our findings show that LRRK2 plays an essential and unexpected role in the regulation of protein homeostasis during aging, and suggest that LRRK2 mutations may cause Parkinson''s disease and cell death via impairment of protein degradation pathways, leading to α-synuclein accumulation and aggregation over time.     

蛛网膜下腔出血剧烈头痛的CT分析

目的　了解SAH后剧烈头痛与CT所示的关系 ,探讨SAH头痛的发病机理。方法　制定头痛程度量表 ,根据CT分析SAH的分布情况 ,结合CSF改变 ,判定出血量和出血部位与头痛程度的关系。结果　CT显示阴性或阳性结果均出现剧烈头痛 ,中等程度以上的出血较少量出血头痛程度明显加重。A区和B区出血头痛程度相同。CSF血性改变程度与头痛严重程度无明显相关。结论　CSF发生血性质变是导致头痛的重要原因 ,头痛程度与出血部位无关。     

减重平板训练对瘫痪后步行障碍患者的影响

目的：观察减重平板训练对脊髓损伤和脑损伤造成的长期下肢瘫痪步行功能的作用。方法：10名男性患者，年龄25－64岁，其中4例脊髓损伤，6例脑血管意外，病程平均1．4年，减重平板训练前均接受不同程度的传统康复训练，功能步行评定为0－1分，接受减重平板训练每天1次，开始减重重量在50％－70％之间，平板速度为0．27m/s，平均20次，所有虱训练前后接受功能步行和一般平衡功能测定，其中7例患者在Kistler压电晶体式三维测力台上进行步态分析。结果：患者经过阶段性减重平板训练，功能性步行评定及站立平衡功能比传统康复治疗前后有十分明显的改善（P＜0．0001，P＝0．0003），其中7例患者的平均步速达0．36m/s，左右足对台压力峰值和支撑时间无明显差异（P＞0．05），结论：减重平板训练对长期瘫痪患者改善步行能力有帮助。     

203例慢性淋巴细胞白血病患者预后相关因素分析

Objective To explore prognostic factors in patients with chronic lymphocytic leukemia (CLL). Methods Two hundred and three CLL patients in our hospital between 2000 to 2007 were retro-spectively reviewed for prognostic factor analysis. Survival was analysed by Kaplan-Meier analysis, univariate analysis by Log-rank test and multivariate analysis by COX regression model. Results With a median follow-up time of 48.0 (3.0 - 156.0) months, the 5-year overall survival (OS) rate was (87.3 ± 2.4) % and 10-year OS rate was (77.4 ± 3.3) %. Forty-eight (23.6%) patients died. Univariate analysis indicated that ad-vanced clinical stage, B symptoms, extranodal involvement, number of lymph node regions involved ≥3, en-larged liver, Hb < 100 g/L, BPC < 100 × 109/L, absolute lymphocyte count (ALC) > 50 × 109/L, atypical cell morphology, progression to stage, non-respons to treatment, complicating infections and secondary cancer or disease transformation were associated with poor prognosis. And on multivariate analysis, lymph node re-gion involvod≥3 and atypical cell morphology were independent poor prognostic factors. Based on the two in-dependent poor prognostic factors, three risk groups were defined: low- (0 factor), intermediate-(one factor) and high-(two factors) groups. The 5 year OS rates were (89.8 ± 3.5) % , (66.4 ～ 7.2) % and (15.0 ±13.8)%, respectively, and the difference between them was statistically. Conclusion The number of lymph node region involved and cell morphology are useful for assessing CLL patients prognosis.