益气活血冲剂对创伤骨折致肾脏损伤的保护作用

目的：探讨益气活血冲剂对创伤骨折所致肾损伤的保护作用。方法：将大鼠或小鼠分为空白组、对照组和实验组。分别于术后不同时期处死动物,测定三组的肾指数、肾中柠檬酸、DNA、RNA含量及^3H-TdR ^3H-UR参入率。结果：创伤骨折引起大鼠肾指数明显下降,^3H-TdR 参入受抑制,DNA含量降低,^3H-UR参入率和RNA含量升高;应用益气活血冲剂可以改善这些变化、促进DNA和RNA的合成。结论：创伤骨折后应用益气活血冲剂可以改善创伤引起的肾脏的变化。     

An indirect treatment comparison of the efficacy of patisiran and tafamidis for the treatment of hereditary transthyretin-mediated amyloidosis with polyneuropathy

Background: Hereditary transthyretin-mediated amyloidosis (hATTR amyloidosis) is a progressive, life-threatening disease. Until recently, tafamidis was the only approved pharmacotherapy. Patisiran significantly improved polyneuropathy and quality of life (QoL) in the phase III APOLLO trial. In the absence of direct comparisons, this analysis aimed to evaluate the comparative efficacy of tafamidis and patisiran in hATTR amyloidosis with polyneuropathy.

Research design and methods: Randomized controlled trial evidence for tafamidis was identified by systematic literature review. Indirect treatment comparisons were performed using the standard pairwise Bucher method for endpoints used in both APOLLO and the tafamidis Fx-005 trial: change from baseline in Neuropathy Impairment Score-lower limbs (NIS-LL), Norfolk QoL-Diabetic Neuropathy questionnaire (QoL-DN), NIS-LL response, and mBMI vs. placebo. Inter-trial population differences were assessed by sensitivity analysis.

Results: The base-case analysis (FAP Stage 1 APOLLO patients vs. intent-to-treat Fx-005 population) suggested patisiran had a greater treatment effect vs. tafamidis for all endpoints, with significant improvements in mean change in NIS-LL (–5.49) and QoL-DN (–13.10) from baseline to Month 18. Similar trends were observed in all sensitivity analyses.

Conclusions: In the absence of direct comparisons, this analysis suggests patisiran has a greater treatment effect than tafamidis in patients with hATTR amyloidosis with polyneuropathy.     

雪芪通肾汤对糖尿病肾病大鼠肾功能及肾脏p38丝裂原活化蛋白激酶mRNA表达的影响

目的探讨雪芪通肾汤对糖尿病肾病大鼠肾功能及p38丝裂原活化蛋白激酶（p38MAPK）mRNA表达的影响。方法将46只大鼠随机分为4组,即正常组10只,模型组、贝那普利组和雪芪通肾组各12只。模型组、贝那普利组、雪芪通肾组用采用链脲佐菌素（STZ）造模。造模成功后,各治疗组均给药24周。取动脉血测肾功能,取左侧肾脏,检测p38MAPKmRNA在肾组织中的表达。结果贝那普利组、雪芪通肾组较模型组的血肌酐（Cr）、尿素氮（BUN）均降低（P〈0．01）。雪芪通肾组较正常组Cr无明显变化（P〉0．05）。半定量分析结果表明贝那普利组、雪芪通肾组p38MAPKmRNA的表达比模型组有所降低（P〈0．01）,比正常组有所升高（P〈0．01）。结论雪芪通肾汤可降低糖尿病大鼠肾脏p38MAPKmRNA表达,保护肾功能,延缓糖尿病肾病进程。     

Altered Regulation of mRNA and miRNA Expression in Epithelial and Stromal Tissue of Keratoconus Corneas

PurposeEvaluation of mRNA and microRNA (miRNA) expression in epithelium and stroma of patients with keratoconus.MethodsThe epithelium and stroma of eight corneas of eight patients with keratoconus and eight corneas of eight non-keratoconus healthy controls were studied separately. RNA was extracted, and mRNA and miRNA analyses were performed using microarrays. Differentially expressed mRNAs and miRNAs in epithelial and stromal keratoconus samples compared to healthy controls were identified. Selected genes and miRNAs were further validated using RT-qPCR.ResultsWe discovered 170 epithelial and 1498 stromal deregulated protein-coding mRNAs in KC samples. In addition, in epithelial samples 180 miRNAs and in stromal samples 379 miRNAs were significantly deregulated more than twofold compared to controls. Pathway analysis revealed enrichment of metabolic and axon guidance pathways for epithelial cells and enrichment of metabolic, mitogen-activated protein kinase (MAPK), and focal adhesion pathways for stromal cells.ConclusionsThis study demonstrates significant differences in the expression and regulation of mRNAs and miRNAs in the epithelium and stroma of Patients with KC. Also, in addition to the well-known target candidates, we were able to identify further genes and miRNAs that may be associated with keratoconus. Signaling pathways influencing metabolic changes and cell contacts are affected in epithelial and stromal cells of patients with keratoconus.     

RNA干扰技术抑制类风湿关节炎成纤维细胞COX-2合成

目的 体外合成针对人环氧化酶-2（hCOX-2）的小分子干扰RNA（siRNA）,并转染人人滑膜成纤维细胞,通过比较不同siRNA抑制COX-2 mRNA表达、COX-2蛋白合成以及下游产物PGE2水平,旨在确定特异性阻断人滑膜成纤维细胞COX-2的siRNA.方法 分离、培养和传代人类风湿关节炎（类风关）滑膜成纤维细胞.设计合成4条针对hCOX-2 mRNA siRNA（1#-4#siRNA）,1条随机序列siRNA.设立1#-4#siRNA和随机序列siRNA组（NC）及未转染对照组（CTL组）.应用Lipofect AMINE 2000将上述siRNA分别转染人滑膜成纤维细胞,4 h后各培养孔加入终浓度为100 nmol/L的佛波酯.转染36 h、48 h后,各组提取蛋白质和总RNA,应用RT-PCR检测hCOX-2 mRNA表达水平,通过Western Blot检测hCOX-2蛋白表达水平.采用ELISA方法检测各组上清液PGE2的水平.结果 转染36 h,PCR结果显示,CTL、NC、1#siRNA、2#RNA、3#RNA、4#siRNA、阳性对照HeLa细胞组hCOX-2 mRNA电泳条带密度值分别为1、0.72、0.3、0.25、0.4、0.04、2.1.Western Blot结果提示上述各组hCOX-2蛋白密度值分别为1、1.04、0.52、0.39、0.9、0和2.48.转染48 h后,各组hCOX-2蛋白条带密度值分别为0.05、0.52、0.51、0.9和0.15.转染24 h、48 h和72 h后,4#siRNA组培养上清液PGE2的水平较其他各组低.结论 4#siRNA能有效抑制人滑膜成纤维细胞COX-2 mRNA表达和COX-2蛋白的合成,且上清液中PGE2水平最低,证实4#siRNA能特异性阻断COX-2在滑膜成纤维细胞表达.     

人喉癌Hep-2细胞膜钾离子通道与RNA编辑酶1的相关性

目的研究人喉癌细胞系Hep-2细胞膜钾离子通道特性及其与RNA编辑酶1(RNA-dependent adenosinedeaminase1,ADAR1)的相关性.方法以Hep-2细胞为研究对象,采用穿孔膜片钳全细胞记录法研究钾离子通道特性,用逆转录聚合酶链反应检测四乙胺(tetraethylammonium,TEA)阻断钾离子通道前后Hep-2细胞ADAR1 mRNA的表达.结果 Hep-2细胞膜的静息膜电位为(-29.8±1.9)mV,在钳制电压-40 mV,阶跃电压在-80～ 80 mV,记录到一种跨膜电流,该跨膜电流具有电压依赖、外向整流特性,该电流在延迟25ms后最大激活,800 ms内不失活,可被阻断剂TEA阻断.Hep-2细胞钾离子通道被TEA阻断前后ADAR1mRNA相对表达量存在显著性差异.结论 Hep-2细胞膜上存在延迟整流钾离子通道,此钾离子通道可能和ADAR1 mRNA的表达密切相关.