Inflammatory and anti-glioma effects of an adenovirus expressing human soluble Fms-like tyrosine kinase 3 ligand (hsFlt3L): treatment with hsFlt3L inhibits intracranial glioma progression.

Glioblastoma multiforme is an intracranial tumor that has very poor prognosis. Patients usually succumb to their disease 6 to 12 months after they are diagnosed despite very aggressive treatment modalities. We tested the efficacy of a potent differentiation and proliferation factor for the professional antigen-presenting dendritic cells (DCs), i.e., Flt3L, for its potential role as a novel therapy for gliomas. We investigated the ability of recombinant adenoviral vectors encoding human soluble Flt3L (hsFlt3L) to improve the survival of Lewis rats bearing intracranial syngeneic CNS-1 gliomas. We show that RAdhsFlt3L can improve survival in a dose-dependent manner. Seventy percent of rats survive when treated with 8 x 10(7) pfu RAdhsFlt3L (P < 0.0005). In addition we demonstrate in both naive Lewis rats and C57BL/6 mice the presence of increased numbers of cells bearing DC markers (OX62 and MHCII, in rats, or CD11C, 33D1, MHCII, and F4/80, but not DEC205, in mice) in sites of brain delivery of RAdhsFlt3L. These results show that expression of hsFlt3L in the brain leads to the presence of cells displaying DC markers. We demonstrate that treatment with hsFlt3L leads to inhibition of tumor growth and significantly increased life span of animals implanted with syngeneic CNS-1 glioma cells. Animals that had survived for long periods, i.e., 6 months, had eliminated the implanted tumors after neuropathological analysis; on the other hand, some of the 3-month survivors still appeared to harbor brain tumors. Our results have profound implications for immune-mediated brain tumor therapy and also suggest the ability to recruit DC-like cells within the brain parenchyma in response to the local expression of Flt3L from adenoviral vectors.     

细胞-支架复合物体内植入修复肌腱缺损的研究与应用

学术背景:肌腱损伤后传统的手术治疗虽然可以减轻患者的痛苦,但其功能重建结果往往不令人乐观。随着组织工程化肌腱的发展,利用组织工程技术修复肌腱缺损已成为一种新的治疗手段。目的:对种子细胞、支架材料、工程化肌腱的构建以及动物实验的方法和检测等研究进展进行综述。检索策略:由作者应用计算机检索Medline,EBSCO以及google scholar引擎中1990-02/2007-05关于组织工程化肌腱的文献,检索词"tissue engineering,engineered tissue,tissue-engineered tendon,scaffold,implantation in vivo,seed cells",检索词被分别组合进行检索,限定文献语言种类为"English"。同时计算机检索维普数据库1990-02/2007-05关于细胞和支架材料的文献,检索词"组织工程,工程化组织,种子细胞,支架,体内植入,组织工程化肌腱",限定文献语言种类为中文。纳入标准:重点选取与构建组织工程化肌腱以及体内植入修复肌腱缺损相关的基础与临床研究。排除标准:关于配子、胚胎和组织工程化骨、血管等的文章。文献评价:①文献来源有RCT,循证医学系统综述、汇总分析、个例报道、经验交流。②共检索到214篇关于细胞和支架材料以及体内植入的文献,其中与本实验关系密切的文献14篇,间接相关45篇,最终纳入35篇符合标准的文献。资料综合:目前运用于组织工程化肌腱的种子细胞主要有间充质干细胞、胚胎干细胞和成纤维细胞,多用聚乳酸、聚羟基乙酸及二者的共聚物作为支架材料。在复合物的培养方面,提出细胞-支架材料复合物体外构建过程中引入动态的机械力牵拉,可使修复后的肌腱具有更好的生物力学强度。组织工程肌腱植入体内后的检测方法除大体观察和组织学检测外,其力学性能检测也是必要的,且分子生物学检测技术也越来越多的应用于研究中。结论:组织工程化肌腱可以成为一种较好修复临床肌腱损伤的方法,但需要探索最适的种子细胞、支架材料、培养条件以及植入体内后的检测方法。     

轮叶党参提取物对酒精性肝损伤的保护作用

目的:探讨轮叶党参提取物保护酒精性肝损伤的作用及其机制。方法:实验于2006-03/12在延边大学基础医学部卫生学教研室完成。选用雄性昆明种小鼠40只,普通级,由延边大学医学部实验动物科提供。按体质量随机分为4组,分别为正常对照组、乙醇组、轮叶党参提取物低剂量组及高剂量组,每组10只。各组小鼠均以每次0.1mL/10g体质量灌胃,正常对照组和乙醇组上午灌胃蒸馏水,轮叶党参提取物低剂量组和高剂量组分别灌胃轮叶党参提取物1g/kg和2g/kg,下午除正常对照组蒸馏水灌胃外,其他各组以食用乙醇灌胃,浓度为450mL/L,连续8周。末次灌胃后禁食12h,颈椎脱臼处死小鼠后迅速取出肝脏,用冷匀浆介质制成10%肝匀浆。检测肝组织中三酰甘油、丙二醛、硒含量及超氧化物歧化酶、谷胱甘肽过氧化物酶活性。结果:实验共纳入昆明中小鼠40只,全部进入结果分析。①轮叶党参提取物对肝脏三酰甘油、丙二醛含量的影响:乙醇组三酰甘油、丙二醛含量明显高于正常对照组(P<0.05),轮叶党参提取物低剂量组和高剂量组三酰甘油、丙二醛含量也明显低于乙醇组(P<0.05),与正常对照组比较,差异不明显(P>0.05)。②轮叶党参提取物对肝脏超氧化物歧化酶、谷胱甘肽过氧化物酶活性及硒含量的影响:低剂量组和高剂量组的超氧化物歧化酶、谷胱甘肽过氧化物酶活性及硒含量均明显高于乙醇组(P<0.05)。结论:轮叶党参提取物可能通过抗氧化、增强自由基及其代谢产物的清除能力,抑制脂质过氧化反应,从而起到对酒精性肝损伤的保护作用。     

肿瘤坏死因子α和吡格列酮对3T3-L1脂肪细胞葡萄糖转运子4表达的影响

目的:观察肿瘤坏死因子α和噻唑烷二酮类药物吡格列酮对3T3-L1脂肪细胞中葡萄糖转运子4(glucose transporter4,GLUT4)mRNA和蛋白表达的影响,并了解其是否有时间依赖性。方法:实验于2005-09/2006-05在安徽医科大学病原微生物分子生物学教研室进行。对3T3-L1细胞进行培养并诱导分化为成熟的3T3-L1脂肪细胞后随机分为对照组、肿瘤坏死因子α组和吡格列酮组3组。肿瘤坏死因子α组和吡格列酮组分别加含20μg/L肿瘤坏死因子α或10-4mol/L吡格列酮培养基培养,提取干预后1,3,6d细胞总RNA和总蛋白做RT-PCR和Western-blot,测定GLUT4mRNA和蛋白的表达,以未加任何药物干预组做对照。结果:①GLUT4mRNA表达:对照组细胞为0.506±0.049;肿瘤坏死因子α组干预后1,3,6d表达低于对照组(0.465±0.039,0.410±0.010,0.320±0.019,F=17.8,P=0.001),并随干预时间呈递减关系;吡格列酮组干预后1,3,6d表达高于对照组(0.544±0.064,0.616±0.065,0.664±0.070,F=4.87,P=0.043),且与干预时间呈正比。②GLUT4蛋白的相对含量:对照组细胞为0.624±0.093;肿瘤坏死因子α组干预后1,3,6d低于对照组(0.549±0.112,0.460±0.111,0.286±0.117,F=7.39,P=0.011),且随干预时间递减;而吡格列酮组GLUT4干预后1,3,6d高于对照组(0.693±0.098,0.750±0.106,0.866±0.074,F=4.0,P=0.048),随干预时间呈递增关系。结论:①肿瘤坏死因子α可降低3T3-L1脂肪细胞中GLUT4mRNA和蛋白表达量,吡格列酮的作用与肿瘤坏死因子α相反,两者作用均呈时间依赖性。②在3T3-L1脂肪细胞中,吡格列酮可能通过增加GLUT4的表达来提高胰岛素敏感性,并可能部分拮抗肿瘤坏死因子α诱导的胰岛素抵抗。     

Hepatitis C viral RNA in clotting factor concentrates and the development of hepatitis in recipients

The polymerase chain reaction (PCR) was used to detect hepatitis C (HCV) viral sequences (HCV-RNA) in clotting factor concentrates that had been stored at 4 degrees C for 1 to 16 years. A total of 43 concentrates were tested, comprising 31 batches of factor VIII, 6 of factor IX, 2 of antithrombin III, 3 of FEIBA, and 1 of factor VII. HCV- RNA was detected in 13 of the 43 batches (30.2%). Concentrates that had not undergone viral inactivation during manufacture were significantly more likely to contain detectable HCV-RNA than concentrates that had been virally inactivated (56.3% v 14.5%, P = .006). HCV sequences were more commonly detected in concentrates made from paid donor plasma than in those made from volunteer donor plasma (44% v 11%, P = .041), and more commonly in virally inactivated concentrates with pre-1989 than with post-1989 expiration dates (50% v 0%, P = .004). Of the four batches of heat-treated products that were HCV-RNA positive, at least three transmitted non-A, non-B hepatitis (NANBH). An association between the presence of HCV-RNA in concentrates and the development of NANBH was demonstrated in nine previously untreated patients on prospective follow-up. HCV-RNA was detected in the concentrates administered to the six patients whose alanine aminotransferase (ALT) abnormalities met the diagnostic criteria for NANBH and who later seroconverted for HCV, but it was not detected in the concentrates administered to the three patients whose ALT abnormalities failed to satisfy the diagnostic criteria and who did not seroconvert. We suggest that the use of this PCR technique to monitor clotting factor concentrates derived from pooled blood may potentially contribute to product safety.