Advanced glycation end products (AGEs) play a critical role in the development of diabetic nephropathy. Reactive oxygen species (ROS) may play a critical role in AGEs induced growth factor expression. In this study, the effects of AGEs on transforming growth factor β1 (TGF-β1), connective tissue growth factor (CTGF) and fibronectin (Fn) mRNA expression and oxidative stress in cultured NRK-49F cells were examined. Methods NRK-49F cells were incubated with medium containing different doses of AGEs (50, 100 or 200μg/ml) for 24 hours, or with AGEs 100 μg/ml for different times (0, 12, 24 or 48 hours). Cells in the serum-free medium or medium containing 25 mmol/L glucose were controls. Cells were treated with 25 mmol/L glucose and 100 μg/ml AGEs for 24 hours to determine the effects between AGEs and glucose. We clarified the role of antioxidant by pretreating cells with N-acetylcysteine (10 mmol/L), ginkgo biloba extract (50 or 100 mg/L) for 24 hours and with 100 μg/ml AGEs for further 24 hours. Alamarblue dye assay was used to analyze cell growth; intracellular ROS generation was measured by flow cytometry; intracellular glutathione by fluorescence spectrophotometry; expressions of TGF-β1, CTGF and Fn mRNA by semiquantitative RT-PCR. Results AGEs significantly increased the expressions of TGF-β1, CTGF, Fn mRNA and intracellular ROS generation, and decreased the glutathion level in NRK-49F cells in dose- and time-dependent manners. High glucose and AGEs together significantly increased the expression of TGF-β1, CTGF and Fn mRNA, compared with AGEs and high glucose separately. Preincubation with N-acetylcysteine or ginkgo biloba extract increased GSH level, suppressed AGEs-induced oxidative stress and TGF-β1, CTGF and Fn mRNA overexpression. Conclusions AGEs can significantly increase expression of TGF-β1, CTGF, Fn mRNA in NRK-49F cells through enhancement of oxidative stress. The accumulation of AGEs may play a pivotal role in the pathogenesis of tubulointerstitial fibrosis in diabetic nephropathy. Suppression of AGEs induced TGF-β1, CTGF and Fn mRNA overexpression in renal fibroblasts through inhibition of oxidative stress may be a mechanism underlying effect of ginkgo biloba extract in diabetic nephropathy. In addition, antioxidant therapy may help prevent AGEs accumulation and its induced damage.

目的建立单核细胞化学吸引蛋白质1（MCP-1）小分子干扰RNA（siRNA）人肾小管上皮细胞株，并检测MCP-1 siRNA对人肾小管上皮细胞（HKC）MCP-1基因的抑制效应。方法针对人MCP-1 mRNA 67、116、142位点设计并合成3对MCP-1 siRNA序列。构建MCP-1特异性siRNA真核表达载体，以脂质体法瞬时转染至HKC。转染24h后分别应用实时定量RT-PCR及Westem印迹技术检测HKC内MCP-1 mRNA、蛋白表达，筛选出抑制效率最高的MCP-1 siRNA。以筛选出的MCP-1 siRNA序列构建慢病毒穿梭质粒。使用慢病毒穿梭质粒进行慢病毒颗粒的包装和生产，得到病毒液并确定其滴度。以病毒液感染HKC，筛选MCP-1 siRNA人肾小管上皮细胞株，并应用实时定量RT-PCR及Westem印迹技术检测HKc内MCP-1 mRNA、蛋白表达。结果成功建市MCP-1特异性siRNA人肾小管上皮细胞株。MCP-1 siRNA稳定转染能下调人肾小管上=皮细胞MCP-1 mRNA（68.49±6.38）%、蛋白（72.97±6.13）%，与对照组比较差异有统计学意义（P<0.01）。结论MCP-1特异性siRNA能高效抑制HKC内MCP-1基因表达。MCP-1 siRNA人肾小管上皮细胞株的建立为MCP-1基因功能及肾问质纤维化防治研究提供实验材料和依据。

目的探讨终末期肾病（End stage renal disease ESRD）患者血浆白细胞介素坞（Interleukin 13 IL-13）水平以及血液透析（Hemodialysis HD）对其的影响，并判断其能否作为评价透析膜生物相容性的指标。方法选择同济大学附属东方医院肾内科30例ESRD患者分为未透析组和维持性血液透析（Maintenance hemodialysis MHD）组，MHD组随机分为纤维素生物膜组（650）和聚砜膜组（F6）各10例，同时设立健康正常者10例为对照组。应用酶联免疫吸附（ELISA）法测定MHD患者透析前、后血浆中IL13水平，以及外周血单个核细胞（Peripherial blood mononuclear cells PBMC）经及未经植物凝集素（Phytohemagglutinin PHA）干预的培养上清中IL-13的水平，同时以半定量逆转录多聚酶链反应（RT-PCR）法检测PBMc中IL-13 mRNA的表达。结果ESRD未透析组及MHD组透析前血浆IL-13水平较正常对照明显升高，MHD组经透析后血浆IL-13水平下降。ESRD未透析患者的PBMC经PHA刺激后IL-13培养上清水平及mRNA表达量均明显降低，650组PBMC经PHA刺激后IL-13培养上清水平及mRNA表达量无明显变化，F6组PBMC经PHA刺激后IL-13培养上清水平及mRNA表达量明显升高。结论①ESRD患者体内处于微炎症状态；②血液透析可改善其炎症状态，从而使IL-13水平下调；③聚砜膜较纤维素生物膜能更好的改善患者的T细胞对抗原的反应能力；④IL-13可作为评价透析膜生物相容性的指标之一。

目的探讨终束期肾病（end stage renal disease，ESRD）患者血浆白细胞舟素18（IL-18）水平以厦不同透析膜时其的影响，井判断其能否作为评价透析膜生物相容性的指标。方法30例ESRD患者分为未遗析组和维持性血液透析（nmintenance hemodialysis，MHO）组，MHD组随机分为纤维素生物膜组（650）和聚砜膜组（F6）各10倒，同时健康者10倒为对照组。应用酶联免疫吸附（ELISA）法测定各组血浆中IL-18水平，以及外周血单个核细胞（pe-rlpherialbkxxlmononudear cells，PBMC）经厦未经脂多糖（lipopolysacchafide，LPS）干预的培养上清中IL-18的水平，同时以半定量逆转录多聚酶链反应（RT PCR）法检测PBMC中IL-18 mRNA的表达。结果ESRD未透析组及MHD组透析前、后血浆IL-18水平较正常时照组明显升高（P<0.05），MHD组透析前、后血浆IL-18水平无明显变化（P>0.05）。ESRD未透析患者的PBMC经LPS刺激后IL-18培养上清水平无明显变化（P>0.05），mRNA表达量则显著降低（P<0.05）；650组透析后PBMC经LPS刺激后IL-18培养上清水平厦mRNA表达量均降低（P<0.05），F6组透析后PBMC经LPS刺激后IL-18培养上清水平及mP，NA表选量均明显升高（P<0.05）。结论（1）ESRD患者体内处于微炎症状态；（2）从血浆IL-18水平变化看，血液透析不能改善ESRD患者体内炎症状态；（3）聚砜膜较纤维素生物膜能改善患者的单核细胞对抗原的反应能力；（4）IL-18可作为评价连析膜生物相客性的指标之一。

血容量监测是血液透析中常用的技术，在评估透析患者干体重及预防透析相关并发症方面发挥了重要的作用，本文就血容量监测的方法及其在血液透析中的应用研究进展进行详细介绍。