+10 Gz重复暴露大鼠脑差异表达基因cDNA文库的构建

目的构建＋10Gz重复暴露大鼠脑差异表达基因的消减cDNA文库。方法本实验用SD大鼠，分别提取暴露组与对照组的总RNA，并分离纯化mRNA，应用抑制性消减杂交技术分离＋10GI重复暴露大鼠脑差异表达基因eDNA片段并建立消减eDNA文库；利用PCR对随机挑选的75个白色菌落进行插入片段的验证，对其中70个克隆进行eDNA斑点杂交验证。结果所构建的eDNA文库扩增后包含约400个白色克隆和100个兰色克隆，随机挑选75个白色克隆入质粒载体后共获得70个阳性克隆。结论应用抑制性消减杂交技术成功构建了＋10Gz重复暴露大鼠脑差异表达基因消减eDNA文库，为进一步筛选和克隆脑损伤相关基因奠定了基础。     

肝动脉解剖变异在中晚期肝癌介入治疗中的意义

目的 探讨放射学肝动脉解剖变异及其在中晚期肝癌介入导管治疗中的临床意义。方法 回顾性分析125例中晚期肝癌肝动脉造影表现及其介入治疗资料。结果 125例中107例(85．6％)有典型肝动脉分布。肝动脉变异18例(14．4％)，其中肝右动脉变异10例(8％)，而又以肝右动脉始于肠系膜上动脉者最多，占6．4％。副肝左动脉变异1例(0．8％)。肝左动脉和肝右动脉同时变异2例(1．6％)。肝总动脉变异3例(2．4％)。胃十二指肠动脉起自肝右动脉和腹腔动脉各1例(1．6％)。对123例成功进行了肝动脉化疗栓塞，2例肝右动脉变异因角度关系超选插管未成功。结论 认识肝动脉解剖变异有助于提高插管的成功率和中晚期肝癌的介入治疗疗效。     

Caffeic acid improves the bioavailability of l‐dopa in rabbit plasma

The impacts of caffeic acid (3,4‐dihydroxycinnamic acid, CA) on the pharmacokinetics of levodopa (L‐dopa) were studied in rabbits. A single dose of 5/1.25 mg·kg^?1 l ‐dopa/carbidopa was administered alone or was co‐administered with three different doses of caffeic acid (2.5, 5, and 10 mg·kg^?1), or a single dose of 5 mg·kg^?1 caffeic acid was administered alone via an intramuscular route to six rabbits each in a crossover treatment protocol. Plasma levels of l ‐dopa, 3‐O‐methyldopa (3‐OMD), caffeic acid, and ferulic acid were determined and subsequently used to calculate their pharmacokinetic parameters. The results indicated that caffeic acid administered at a dose of 10 mg·kg^?1 decreased about 22% of the peripheral formation of 3‐OMD and about 31% of the C_max of 3‐OMD. In addition, the metabolic ratios (MR, AUC of 3‐OMD/AUC of L‐dopa) decreased by about 22%. Results also indicated that caffeic acid significantly decreased the proportion of 3‐OMD (p < 0.05). In contrast, the parameters of neither caffeic acid nor ferulic acid were significantly affected by l ‐dopa/carbidopa. In conclusion, caffeic acid at a dose of 10 mg·kg^?1 can significantly affect the COMT metabolic pathway of L‐dopa. Copyright © 2009 John Wiley & Sons, Ltd.     

胶束液相色谱法测定动物肌肉中三聚氰胺残留量

目的 建立胶束HPLC测定动物肌肉中三聚氰胺残留量的分析方法。方法 样品在酸性条件下，用乙 腈-磷酸盐缓冲液提取。以胶束溶液作为流动相，用HPLC测定，外标法定量。结果 线性范围为（0.1～10.0）mg/L，相关系数为0.9996。动物肌肉在（0.5～5.0）mg/kg添加水平范围内，回收 率在80%～110%。相对标准偏差〈10%。方法的最低检出限0.5mg/kg。结论该法实用、准确，为动物肌肉中 三聚氰胺残留量检测提供了可行的方法。     

Protein kinase C�� expression in breast cancer as measured by real-time PCR, western blotting and ELISA

The protein kinase C (PKC) family of genes encode serine/threonine kinases that regulate proliferation, apoptosis, cell survival and migration. Multiple isoforms of PKC have been described, one of which is PKCδ. Currently, it is unclear whether PKCδ is involved in promoting or inhibiting cancer formation/progression. The aim of this study was therefore to investigate the expression of PKCδ in human breast cancer and relate its levels to multiple parameters of tumour progression. Protein kinase Cδ expression at the mRNA level was measured using real-time PCR (n=208) and at protein level by both immunoblotting (n=94) and ELISA (n=98). Following immunoblotting, two proteins were identified, migrating with molecular masses of 78 and 160 kDa. The 78 kDa protein is likely to be the mature form of PKCδ but the identity of the 160 kDa form is unknown. Levels of both these proteins correlated weakly but significantly with PKCδ concentrations determined by ELISA (for the 78 kDa form, r=0.444, P<0.005, n=91 and for the 160 kDa form, r=0.237, P=0.023, n=91) and with PKCδ mRNA levels (for the 78 kDa form, r=0.351, P=0.001, n=94 and for the 160 kDa form, r=0.216, P=0.037, n=94). Protein kinase Cδ mRNA expression was significantly higher in oestrogen receptor (ER)-positive compared with ER-negative tumours (P=0.007, Mann–Whitney U-test). Increasing concentrations of PKCδ mRNA were associated with reduced overall patient survival (P=0.004). Our results are consistent with a role for PKCδ in breast cancer progression.     

尼尔雌醇和左炔诺孕酮对人成骨肉瘤 MG-63细胞株雌激素受体亚型的作用

目的：观察不同浓度的尼尔雌醇(NYL)和左炔诺孕酮(LNG)对人成骨肉瘤MG-63细胞株ERα和ERβ表达的作用，探讨旁分泌效应对ERα和ERβ基因表达的影响。方法：人成骨肉瘤MG-63细胞株分为（1）空白对照组：用含1% BSA无血清MEM培养液培养48 h；（2）阳性对照组：培养液中加入10-8 mol/L的17β-estradiol (E2) 培养48 h；（3）药物处理组：培养液中分别加入10-10，10-8，10-6 mol/L的NYL或LNG培养48 h；（4）药物处理+换液组：培养液中分别加入10-10 mol/L的NYL或10-8 mol/L LNG培养48 h，每12 h更换新的含有相应药物的培养液。用半定量RT-PCR检测各组细胞ERα和ERβmRNA的表达。结果：NYL及LNG均能诱导ERα和ERβ亚型mRNA表达上调，诱导ERαmRNA表达的最强作用浓度均为10-6 mol/L。NYL，LNG对ERβ mRNA表达最强作用浓度分别为10-10，10-8 mol/L。每12 h更换干预培养液对ERα表达无影响，但可抑制ERβ的表达。结论：NYL和LNG均可诱导MG-63细胞株ERα和ERβ mRNA表达上调，且2种亚型的表达存在相互制约关系；在NYL和LNG诱导ER亚型表达上调过程中ERβ的表达可能受到旁分泌作用的影响。