相关生物因子对体外培养关节软骨细胞促增殖作用的比较研究

目的　筛选关节软骨组织工程种子细胞优化获取的高效促增殖剂。　方法　采用培养细胞3H -胸腺嘧啶脱氧核苷 (3H -TdR)掺入实验、四甲基偶氮唑蓝 (MTT)代谢检测及Ⅰ、Ⅱ型胶原mRNA逆转录聚合酶链反应 (RT -PCR)分析方法 ,比较一定浓度维生素C(vitaminC ,VC) +骨形态发生蛋白 2 (bonemorphogeneticprotein 2 ,BMP2 )、类胰岛素生长因子 1(insulin -likegrowthfac tor1,IGF1 ) (体积分数为 2 %的血清培养时 )及碱性成纤维细胞生长因子 (basicfibroblastgrowthfactor,bFGF)、转化生长因子 - β1 (transforminggrowthfactor- β1 ,TGF - β1 ) (体积分数为 10 %的血清培养时 )对适宜密度接种、体外单层传代培养兔关节软骨细胞的促增殖作用。　结果　适宜浓度VC +BMP2 、IGF1 及bFGF、TGF - β1 均可促进体外传代培养关节软骨细胞的分裂增殖。增殖后的培养细胞仍具有其特异性表型表达 ,而IGF1 培养的细胞则有向成骨样细胞分化的趋向。其综合效能依次为 :VC +BMP2 >bFGF >TGF - β1 >IGF1 。　结论　一定浓度VC +BMP2 及bFGF对适宜密度接种、体外单层传代培养关节软骨细胞具有显著的促增殖作用 ;其中VC +BMP2 具有经济、高效的优点。     

微波辐射对培养的hTERT-RPE1细胞的应激损伤

目的：观察2450MHz微波辐射对培养的hTERT—BPEl细胞的应激损伤。方法：体外培养的人视网膜色素上皮细胞系细胞，分别在强度为10、20、30mW／cm2的微波下辐射1h，辐射后，用光学显微镜和透射电镜观察各组细胞的形态及变化；台盼蓝染色检测细胞存活率；测定细胞内抗氧化物酶SOD和GSH—Px的活性和脂质过氧化产物MDA的含量；另外，继续培养各组细胞，观察微波辐射对细胞增殖的影响。结果：微波辐射后，细胞存活率、细胞内抗氧化物酶SOD、GSH—Px活性与对照组相比显著降低；而过氧化产物MDA含量与对照组相比显著升高，其降低或升高的幅度与微波辐射的强度的大小有关。结论：2450MHz微波辐射可引起培养的hERT—BPEl细胞的应激损伤。     

Tetrahydroporphyrin-tetratosylat (THPTS): A near-infrared photosensitizer for targeted and efficient photodynamic therapy (PDT) of human bladder carcinoma. An in vitro study

BackgroundEfficacy of PDT in muscle-invasive bladder cancer is hampered by low tissue penetration of most photosensitizers by short excitation wavelength. THPTS is excitable at near-infrared (760 nm) allowing tissue penetration up to 15 mm. We examined the cellular effects of THPTS-PDT in human bladder cancer cells.Material and methodsWe used four human transitional carcinoma cell lines, epithelial bladder progenitors (HBLAK) and bladder smooth muscle cells (HBSMC). We used flow cytometry to examine pharmacokinetics of THPTS, confocal laser scanning microscopy to analyze subcellular localization and production of reactive oxidative species (ROS), examined cytotoxicity and cell death pathways (qRT-PCR).ResultsTotal uptake varied between cell lines and was significantly high in HBLAK and HBSMC. Lysosomal localization was mainly seen in cancer cells and HBLAK, while THPTS was distributed throughout the cytoplasm in HBSMC. Significant ROS production was detected 30 min after THPTS-PDT. Growth arrest occurred within 4 h and resulted in apoptotic and necrotic cytotoxicity after 24 h. Cytotoxicity was dose-dependent and specifically high in cancer cells and HBLAK and significantly low in HBSMC.ConclusionTHPTS-PDT induces cellular mechanisms leading to cellular growth arrest, apoptosis and necrosis in human bladder cancer cells. These effects are only partly dependent on the total amount of THPTS uptake and rather dependent on its subcellular compartmentalization. HBSMC are hardly affected by THPTS-PDT confirming tumor specificity and safety. THPTS is a promising new photosensitizer with the unique advantage of deep tissue penetration allowing the treatment of solid tumors and warranting further animal studies.     

胎儿骨髓间充质干细胞的体外培养及其生物学特性的研究

目的：确定人胎儿骨髓间充质干细胞(Fetal mesenchymal stem cells，FMSCs)体外分离和培养的方法，观察其生物学特性，建立稳定的MSCs体外培养扩增体系。方法：通过密度梯度离心、贴壁筛选法分离培养人FMSCs，观察细胞的形态与超微结构，并研究其增殖及生长特征，应用流式细胞术测定细胞周期和CD14、CD29、CD34、CD44、CD45的表达。结果：在体外培养条件下人FMSCs贴壁生长，为成纤维细胞样，CD14／CD34／CD45阴性，CD29／CD44阳性，核浆比大，细胞周期检测C0／C1期约占95％，具有原始细胞的特征。结论：体外培养可获得较均一的FMSCs，形态单一、生长稳定、增殖性较强，可以用于细胞治疗的进一步研究。     

体外培养半月板纤维软骨细胞增殖和胶原表达特性的研究

目的　研究体外单层培养条件下猪半月板纤维软骨细胞增殖和胶原的表达,为构建自体组织工程化半月板奠定基础。　方法　采用改良的Klagsbrun法,从8只45d龄长枫杂交仔猪的半月板消化获取细胞进行体外单层培养。形态学观察、群体倍增时间(populationdoubletime,PDT)、增殖速度趋势图、原位杂交及免疫细胞化学染色法作为评定指标。　结果　半月板组织的平均细胞获得率为(2.31±0.66)×10     

体外培养人胎肝细胞与永生化L-02肝细胞的生物学性状比较

目的 对比体外培养的人胎肝细胞与L-02肝细胞形态学及增殖分化特征的异同，初步评价其用于移植的可行性，探讨介入性肝细胞移植术的理想供体来源。方法 分离培养14～24周的引产胎儿肝细胞，放射免疫法测定上清液中甲胎蛋白(AFP)与白蛋白(ALB)含量，免疫细胞化学法检测细胞角蛋白19(CK—19)的表达。同法检测传代培养L-02肝细胞的蛋白表达。结果人胎肝细胞分离活率达95％，在体外存活最长达3周，可同时检测到AFP、ALB及CK—19表达，其中ALB分泌峰值42．06μg／ml；L-02肝细胞增殖迅速，AFP与CK-19表达阴性，ALB表达在10μg／ml水平，部分细胞多次传代后发生形态变异，ALB表达缺失。结论人胎肝细胞具有潜在的双向分化能力，是肝细胞移植较为合适的供体来源；L-02肝细胞不适用于移植。     

Methamphetamine induces an increase in cell size and reorganization of myofibrils in cultured adult rat cardiomyocytes

To investigate the direct effects of methamphetamine (MAP) on cardiac lesions seen in MAP abusers, isolated adult rat ventricular cardiomyocytes (ARCs) were exposed to MAP (0.05–1.0 mM) in medium 199 containing 10% fetal calf serum. Isolated ARCs attached to laminin-coated substrata and began to spread into polygonal shapes with pseudopodia at day 6 in normal culture. However, the cell attachment and spreading were inhibited by exposure to MAP (0.5 and 1.0 mM) for the first 7 days in culture. On the other hand, exposure to MAP (0.05 and 0.1 mM) for 7 days after a 6-day period of normal culture, led to a larger cross surface area of cells with more abundant actin bundles compared to control cells (p < 0.05). This development of spreading area resembled that of norepinephrine-treated ARCs. In addition, immunoreactive atrial natriuretic peptide (ANP) granules developed and accumulated around the nuclear region of ARCs exposed to MAP and the number of ANP positive cells tended to increase in a dose-dependent manner. These results suggest that chronic exposure to a high concentration of MAP may directly inhibit development of ARCs in culture and that a continuous exposure to a low concentration of MAP may facilitate the development of cellular hypertrophy. Therefore, hypertrophied cardiomyocytes in MAP abusers may be provoked by multifactorial incidents of direct and indirect actions of MAP. Received: 1 February 1999 / Received in revised form: 1 July 1999     

NDV对体外培养的肿瘤细胞杀伤性研究

目的：探讨ＮＤＶ病毒体外对肿瘤细胞的杀伤性。方法：人的ＳＧＣ－９７０１和ＳＭＭＣ－７７２１肿瘤细胞为靶细胞,观察了ＮＤＶ对其直接杀伤作用。结果：ＮＤＶ病毒能作用于瘤细胞,并最终使肿瘤细胞死亡。结论：ＮＤＶ病毒是一种有效的肿瘤细胞溶解剂,在体外具有明显的直接杀伤瘤细胞作用。     

5-羟色胺及其受体在人肝癌细胞系HCC-9204上的免疫组织化学定位

目的：探讨５－羟色胺及其受体与人肝细胞性肝癌之间的关系。方法：应用免疫组织化学ＳＡＢＣ法对培养的人肝癌细胞系ＨＣＣ－９２０４细胞爬片进行染色。结果：５－羟色胺及其受体在肝癌细胞上皆有表达,其分布范围大体一致。结论：５－羟色胺可能参与人肝细胞性肝癌的增殖,ＨＣＣ０－９２０４细胞系可以作为研究５－羟色胺与肝癌关系的一种有用材料。     

胆石症患者血清,胆汁,胆石相关元素分析和胆石成因的研究

利用火焰原子吸收法，对２７例胆石症患者血清、胆汁和胆石的相关元素进行测定，结果表明，胆石症患者血清钙明显高于正常人，而血清铜则明显低于正常人（Ｐ＜０．０１）；胆汁中钙、铜、钠、镁等元素明显低于对照组（Ｐ＜０．０１）；胆色素结石组钙、钠、镁等元素亦明显高于胆固醇结石组，说明血清钙、铜代谢异常及胆汁中钙、铜、钠、镁等元素含量的改变与胆石形成有密切关系。