内皮素A受体拮抗剂BQ123对前列腺癌PC-3M细胞增殖与凋亡的影响

目的:观察内皮素A受体拮抗剂BQ123对转移性前列腺癌(PCa)PC-3M细胞株增殖与凋亡的影响,初步探讨内皮素A受体拮抗剂对转移性PCa细胞的体外抗肿瘤效应。方法:实验分为两组,对照组为PC-3M细胞及含灭活小牛血清的F12/RMPI1640培养液;实验组为PC-3M细胞加入等量的1μmol/LBQ123。采用四甲基偶氮唑蓝(MTT)比色法、划痕损伤实验、细胞迁移实验观察BQ123对人PCaPC-3M细胞株增殖的抑制效应,利用Annexin V-FITC/PI染色和流式细胞术检测BQ123诱导PC-3M细胞凋亡的作用及其与细胞周期的关系。结果:MTT比色法结果显示BQ123随着作用时间的增加,对PC-3M的抑制率分别为24h22.32%、48h44.88%和72h64.47%,表现出良好的时效关系(P<0.05),划痕损伤实验结果提示:实验组PC-3M细胞的迁移距离分别为12h(103.42±75.63)μm、24h(243.75±121.53)μm和48h(422.07±36.01)μm,均低于对照组的12h(162.93±19.87)μm、24h(317.19±43.19)μm和48h(692.74±40.84)μm(P<0.05)。细胞迁移实验中实验组穿入下室的PC-3M细胞为(79.2±9.58)个,亦明显少于对照组的(92.6±5.94)个(P<0.05);Annexin V-FITC/PI双染色法的结果显示对照组PC-3M细胞凋亡率为(9.38±1.37)%,实验组PC-3M细胞凋亡率为(15.03±0.93)%,差异有统计学意义(P<0.05),细胞周期实验的结果显示实验组各期细胞比例与对照组相比差异不显著(P>0.05)。结论:BQ123对PCaPC-3M细胞株的生长、迁移和侵袭能力均有明显的抑制作用,且可以诱导PC-3M细胞凋亡,可为PCa治疗的研究提供一种新的思路。     

123I-2-iodo-tyrosine, a new tumour imaging agent: human biodistribution, dosimetry and initial clinical evaluation in glioma patients

Purpose ¹²³I-2-iodo-tyrosine (¹²³I-2IT) has been identified as a promising new amino acid tracer in animals. Uptake is mediated by LAT1 transport, which is increased in tumour cells. In this study we present the human biodistribution and first clinical results in glioma patients. Methods For the biodistribution study, six male volunteers received 60–95 MBq ¹²³I-2IT. Whole-body scans and blood and urine samples were obtained up to 24 h after injection; dosimetry was calculated using OLINDA 1.0 software. Initial clinical evaluation of ¹²³I-2IT SPECT was performed in 35 patients with suspected or known glioma, either as primary diagnosis or for detection of recurrence. Tumour-to-background (T/B) ratios were calculated for semi-quantitative analysis. The results were correlated with clinical and MRI follow-up data or histology. Results ¹²³I-2IT showed both renal and intestinal clearance. Bladder (0.12 mGy/MBq) and small intestine (0.03 mGy/MBq) received the highest absorbed doses. The effective dose equivalent and effective dose were estimated at 0.020 and 0.016 mSv/MBq, respectively. In patients, ¹²³I-2IT SPECT did not differentiate between neoplastic and non-neoplastic lesions after an indeterminate MRI. In follow-up of known glioma, 13/15 patients with disease recurrence had increased T/B values (range 1.39–3.91). Out of seven recurrence-negative patients, two showed an important increase in T/B, in one case due to radionecrosis (T/B 1.59) and in the other probably due to residual but stable disease (T/B 2.07). Conclusion ¹²³I-2IT has a favourable biodistribution for a tumour imaging agent. It shows increased uptake in central nervous system glioma and is potentially useful in the follow-up of glioma patients. M. Keyaerts is an “aspirant” of the FWO-Vlaanderen.     

Neuronal imaging using SPECT

Background ¹²³I-metaiodobenzylguanidine (MIBG) myocardial scintigraphy is one of only a few methods available for objective evaluation of cardiac sympathetic function at a clinical level. Disorders in cardiac sympathetic function play an important role in various heart diseases, and MIBG provides an abundance of useful information for evaluation of disease severity, prognosis, and therapeutic effects; this information is of particular value in patients with heart failure, ischemic heart diseases, or arrhythmic disorders. On the other hand, the quantitative indices for MIBG differ between institutions, and evidence has not been sufficiently well established for MIBG, compared with myocardial perfusion imaging, in ischemic heart diseases. Review In view of these difficulties, this review provides fundamental information regarding MIBG, its usefulness for various diseases and future difficulties.     

Age-related decline in nicotinic receptor availability with [(123)I]5-IA-85380 SPECT

Human postmortem studies have reported decreases with age in high affinity nicotine binding in brain. We investigated the effect of age on β₂-containing nicotinic acetylcholine receptor (β₂-nAChR) availability in eight brain regions of living human subjects using the ligand [¹²³I]5-IA-85380 ([¹²³I]5-IA) and single photon emission computed tomography (SPECT). Healthy, nonsmokers (N = 47) ranging in age from 18 to 85 were administered [¹²³I]5-IA using a bolus plus constant infusion paradigm and imaged 6–8 h later under equilibrium conditions. The effect of age on regional β₂-nAChR availability (V_T, regional brain activity/free plasma parent, a measure proportional to the binding potential) was analyzed using linear regression and Pearson's correlation (r). Age and regional β₂-nAChR availability were inversely correlated in seven of the eight brain regions analyzed, with decline ranging from 32% (thalamus) to 18% (occipital cortex) over the adult lifespan, or up to 5% per decade. These results in living human subjects corroborate postmortem reports of decline in high affinity nicotine binding with age and may aid in elucidating the role of β₂-nAChRs in cognitive aging.     

体外诱导脐血单核细胞向破骨样细胞的分化

背景：正畸牙齿移动的基础是牙周组织的改建,其中破骨细胞性骨吸收是牙齿移动的第一步,应力作用下有关破骨细胞分化和功能成熟的信号转导通路,以及牙周膜细胞和破骨细胞之间的关系是目前国内外研究的热点之一。 目的：拟建立人破骨样细胞体外培养的简便方法,观察骨吸收刺激因子对破骨样细胞分化、增殖和功能的影响。 设计、时间及地点：细胞学体外对照观察,于2007-10/2008-05在西安交大口腔医院中心实验室完成。 材料：脐带血来源于非高危妊娠的健康产妇,新鲜牛股骨由西安交通大学动物实验中心提供,用于制备100~200 μm厚的骨片, 1α,25-(OH)2D3、巨噬细胞集落刺激因子、前列腺素E2等骨吸收刺激因子均为Sigma公司产品。 方法：无菌条件下收集脐带血,Ficoll液分离后吸取呈云雾状的白膜层,离心弃上清,加入α-MEM培养液重悬,调整脐血单核细胞浓度为1×109 L-1,接种于预置盖玻片和骨片的24孔培养板中,1.0 mL/孔,设立空白对照组、10-8 mol/L及10-7 mol/L 1α,25-(OH)2D3组、巨噬细胞集落刺激因子组、1α,25-(OH)2D3+前列腺素E2组,培养7 d。 主要观察指标：倒置显微镜观察细胞生长形态,TRAP染色法观察破骨样细胞的形成,甲苯胺蓝染色观察骨吸收陷窝情况,以TRAP染色(+)、细胞核≥2个的细胞为破骨样细胞进行计数。 结果：培养3 d后,空白对照组细胞形态及数量无明显变化,各诱导组单核细胞出现融合趋势;7 d时空白对照组出现少量的破骨样细胞,核的数目2~3个,各诱导组可见大量多核破骨样细胞,核的数目3~20个不等。诱导后光镜下可见胞浆呈红色、胞核呈淡黄色的TRAP(+)破骨样细胞,尤其是 10-8 mol/L 1α,25-(OH)2D3组可见含14个核的强阳性破骨样细胞,且胞体较大。各组均尚未形成骨吸收陷窝。与空白对照组比较,各诱导组破骨样细胞数量均明显增多（F=9.78,P < 0.01）;与10-8 mol/L 1α,25-(OH)2D3组比较,10-7 mol/L 1α,25-(OH)2D3组破骨样细胞数量无明显变化（P > 0.05）,巨噬细胞集落刺激因子组及1α,25-(OH)2D3+前列腺素E2组破骨样细胞数量均明显减少（F=7.46,P < 0.01）。 结论：脐血单核细胞经骨吸收刺激因子体外诱导培养后,可分化为TRAP(+)的多核破骨样细胞,其中10-8 mol/L 1α,25-(OH)2D¬具有最强的生物学效应。     

治疗型关节炎护膝对实验性骨关节炎组织形态学的影响

背景：治疗型关节炎护膝具有通经活络、疏导瘀滞、行气活血的作用,已证实是防治膝骨性关节炎行之有效的医疗仪器。 目的：通过观察兔膝骨性关节炎关节软骨组织的病理变化,探讨治疗型关节炎护膝的治疗作用。 设计、时间及地点：随机对照动物观察,于2006-01/10在解放军南京军区福州总医院动物实验中心及病理中心完成。 材料：健康6月龄日本大耳白兔54只,由无锡市惠山江南动物实验中心提供。治疗型关节炎护膝由福建中医学院和福州大学生物医学研究所共同研制,包括疏密波、热软膜程序,两种程序联合为温热动态干扰变频疏密波。微波仪产地日本。 方法：取10只兔作为正常对照组,剩余44只采用改良Hulth法建立膝骨性关节炎模型,造模后随机分为模型对照组9只、微波对照组9只、疏密波组9只、热软膜组8只、疏密波+热软膜组9只。造模后7 d开始每日强迫兔活动30 min,之后正常对照组、模型对照组不进行任何干预,其余组兔固定后分别对应给予微波仪治疗、疏密波治疗、热软膜治疗、疏密波+热软膜联合治疗,30 min/次,1次/d,连续16周。 主要观察指标：通过CR片、光镜和电镜观察膝关节组织形态学变化。 结果：干预16周后,CR片显示正常对照组关节间隙正常,关节面平整光滑,无骨赘;模型对照组内侧间隙明显变窄,有明显骨赘。光镜下正常对照组关节软骨四层结构清晰可辨,软骨细胞排列整齐呈柱状,染色质分布均匀,细胞核清晰;模型对照组关节软骨层变薄,部分区域软骨细胞核固缩、坏死,软骨细胞排列紊乱,四层结构不易分辨。电镜下正常对照组软骨细胞呈椭圆形,细胞及胞膜完整,包质内可见丰富的粗面内质网、高尔基体、线粒体,细胞核完整,染色质分布均匀;模型对照组软骨细胞明显固缩且外形不规则,细胞周晕消失,胞浆内细胞器凝成高电子密度的片状物不易分辨,细胞核不 规则,染色质浓聚,散裂于核中。疏密波+热软膜组在病理程度上的改变明显轻于模型对照组,软骨表面裂纹少,胶原纤维结构基本完整,固缩的软骨细胞少,虽可见部分退变的软骨细胞,但部分软骨细胞具有较多的细胞器,某些区域形成软骨 细胞簇。微波对照组、单纯疏密波组、单纯热软膜组膝关节组织形态学变化介于模型对照组和疏密波+热软膜组之间。 结论：治疗型关节炎护膝的疏密波+热软膜程序能延缓膝关节软骨宏观形态、软骨细胞及软骨基质的退变,效果优于微波仪治疗或单纯的疏密波、热软膜治疗。     

聚乳酸-羟基乙酸包载眼镜蛇毒细胞毒素缓释微球的表征及体外释放行为

背景：眼镜蛇毒细胞毒素具有强烈的细胞毒活性,但缺乏特异性,全身用药可导致严重毒副作用,而采用缓释载体包载进行间质化疗可达到提高肿瘤局部治疗效应,并且减轻全身毒性。 目的：制备眼镜蛇毒细胞毒素-聚乳酸-羟基乙酸微球,观察其一般性质和体外释药特性。 设计、时间及地点：观察性实验,于2007-12/2008-05在福建医科大学医药生物工程中心完成。 材料：聚乳酸-羟基乙酸、聚乙烯醇由中国科学院成都有机化学有限公司提供,广东产中华眼镜蛇毒。 方法：采用分子筛、离子交换分离,反相疏水高效液相色谱方法纯化细胞毒素,MTT法检测细胞毒活性,复乳-溶剂挥发法制备载药微球。 主要观察指标：扫描电镜观察载药微球的表面形态,激光粒径仪测微球粒径,计算包封率、载药量、体外释放周期。 结果：纯化的眼镜蛇细胞毒素具有明显的细胞毒作用,对HepG2细胞12,24 h的IC50分别为1.43,1.12 mg/L。复乳法制备微球表面光滑圆整,粒径2~8 μm,包封率和载药率分别为(74.10±9.92)%和(0.72±0.09)%,21 d药物累积释放63.3％,释放细胞毒素保持较好的生物学活性。 结论：采用复乳-溶剂挥发法可制备具有较高包封率、良好缓释效果、保持完整生物学活性的眼镜蛇毒细胞毒素-聚乳酸-羟基乙酸微球。     

组织工程化类金刚石膜复合材料与人血管内皮细胞的相容性研究

目的：验证纳米相类金刚石薄膜复合材料与人血管内皮细胞相容性,为组织工程化机械瓣膜材料的构建提供依据。 方法：利用脉冲激光沉积法在人工心脏机械瓣膜上沉积纳米相类金刚石薄膜,将人脐静脉血管内皮细胞与纳米相类金刚石薄膜复合材料体外复合培养。倒置显微镜及扫描电镜观察细胞在材料表面的生长、附着情况;MTT 法检测细胞在材料上增殖情况;同时分别测定人脐静脉血管内皮细胞在类金刚石薄膜材料和空白对照组中一氧化氮及前列环素分泌水平,以评价其活性。 结果：人脐静脉血管内皮细胞能在纳米相类金刚石薄膜复合材料上良好地黏附、增殖、生长。人脐静脉血管内皮细胞分泌一氧化氮和前列环素水平在类金刚石薄膜材料和空白对照组没有显著性差异(P > 0.05),说明纳米相类金刚石薄膜材料对人脐静脉血管内皮细胞的活性没有影响。 结论：纳米相类金刚石薄膜复合材料具有良好的细胞相容性,有可能作为组织工程化机械瓣膜材料。     

腹膜透析管材料特征及相关性感染

腹膜透析是急、慢性肾衰竭的主要替代疗法之一,与血液透析相比,它具有操作简单,可自行透析和最大限度保护残存肾功能的特点,目前已在国内外广泛开展。腹膜透析相关性感染是影响患者透析疗效和病死率的主要原因。资料显示,腹膜透析相关性感染与多种因素有关,包括操作方法、患者的营养状况、家居环境、文化程度及肠道因素等,而腹膜透析管材料特征所导致的感染越来越受到重视。腹膜透析管类型、临床置管操作方法的不统一以及熟练程度都会影响腹膜透析患者的治疗效果及远期预后。