Ammonium accumulation and chemokine decrease in culture media of Gcdh−/− 3D reaggregated brain cell cultures

Glutaric Aciduria type I (GA-I) is caused by mutations in the GCDH gene. Its deficiency results in accumulation of the key metabolites glutaric acid (GA) and 3-hydroxyglutaric acid (3-OHGA) in body tissues and fluids. Present knowledge on the neuropathogenesis of GA-I suggests that GA and 3-OHGA have toxic properties on the developing brain.We analyzed morphological and biochemical features of 3D brain cell aggregates issued from Gcdh^?/? mice at two different developmental stages, day-in-vitro (DIV) 8 and 14, corresponding to the neonatal period and early childhood. We also induced a metabolic stress by exposing the aggregates to 10 mM l-lysine (Lys).Significant amounts of GA and 3-OHGA were detected in Gcdh^?/? aggregates and their culture media. Ammonium was significantly increased in culture media of Gcdh^?/? aggregates at the early developmental stage. Concentrations of GA, 3-OHGA and ammonium increased significantly after exposure to Lys. Gcdh^?/? aggregates manifested morphological alterations of all brain cell types at DIV 8 while at DIV 14 they were only visible after exposure to Lys. Several chemokine levels were significantly decreased in culture media of Gcdh^?/? aggregates at DIV 14 and after exposure to Lys at DIV 8.This new in vitro model for brain damage in GA-I mimics well in vivo conditions. As seen previously in WT aggregates exposed to 3-OHGA, we confirmed a significant ammonium production by immature Gcdh^?/? brain cells. We described for the first time a decrease of chemokines in Gcdh^?/? culture media which might contribute to brain cell injury in GA-I.     

中西医治疗糖尿病周围神经病变研究进展

对于糖尿病周围神经病变的治疗,中医以"络以通为用"为治疗原则,在改善临床症状,阻止DPN进一步恶化,减少截肢率方面,都有很好的疗效。西医可以有效控制血糖、血压,弥补了中药不能快速降糖降压之不足,对防止DPN的发生和发展起了重要作用。中西医结合治疗具有一定互补优势,疗效确切,但其作用机制尚不明确,需进一步加强实验研究,明确中医药作用机理,提高临床疗效。     

龙血竭胶囊合九华膏对环状混合痔术后创面促愈作用及对新生血管形成的影响

目的:观察龙血竭胶囊合九华膏对环状混合痔术后恢复的疗效,及其对创面和血清血管内皮生长因子的影响。方法:将190例患者根据入院先后顺序,随机按数字表法分为对照组和观察组各95例。两组患者均行分段多层次弧切缝合加注射术,术后常规进行抗生素静脉滴注预防感染;并内服芪蓉润肠口服液,20 mL/次,3次/d,便后采用苦参汤伤口清洗。对照组采用马应龙麝香痔疮膏涂抹肛门,以无菌纱布覆盖创面,1 g/次,1~2次/d。观察组以龙血竭胶囊合九华膏涂抹肛门,以无菌纱布覆盖创面,1 g/次,1~2次/d。两组疗程均为10 d。记录术后第3,10天创面的疼痛、水肿、创面渗液、创面面积和创面肉芽组织情况,记录创面愈合时间;评价术后10 d创面毛细血管含量,采用免疫组化评价创面血管内皮生长因子(VEGF),碱性成纤维生长因子(b FGF)表达情况;检测治疗前后血清血小板源性生长因子(PDFG),血管内皮生长因子(VEGF),碱性成纤维生长因子(b FGF)。结果:术后10 d,观察组临床疗效总有效率为91.58%,优于对照组的77.89%(χ~2=6.877,P0.01);观察组在术后3 d和10 d疼痛、水肿、创面渗液、创面面积和创面肉芽组织评分均低于同期对照组(P0.01);观察组创面平均愈合时间为(11.5±2.1)d,低于对照组的(13.7±2.5)d,比较差异有统计学意义(P0.01);观察组创面毛细血管数目多于对照组(P0.01),观察组创面VEGF和b FGF表达的MOD值高于对照组(P0.01);治疗后观察组患者血清PDFG,VEGF和b FGF水平均高于对照组,差异有统计学意义(P0.01)。治疗过程中未发现与龙血竭联合九华膏相关不良反应。结论:采用龙血竭胶囊合九华膏治疗环状混合痔术后创面的治疗,能促进创面的修复,缩短愈合时间,其作用机制可能与调节血清和创面VEGF,PDFG,b FGF等因子表达,促进创面毛细管血管形成,促使肉芽组织生长与修复有关。     

渐进式规范化岗前培训提升入职护士临床技能的研究

针对新入职护士规范化岗前培训对快速提升临床技能的研究状况、进展以及影响新入职护士规范化岗前培训的因素进行综述,以期为新入职护士的渐进式规范化培训的进一步研究提供参考。     

不同浓度bFGF对犬骨髓基质细胞增殖、分化影响的实验研究

目的：研究不同浓度的碱性成纤维细胞生长因子（basic fibroblast growth factot,bFGF）对体外培养的beagle犬骨髓基质细胞（bone marrow stromal ceils，BMSCs）增殖和分化的影响。方法：体外培养第2代BMSCs的培养液中分别加入不同浓度（25、50、100、200ng／mL）的bFGF，连续6d倒置显微镜下动态观察BMSCs的形态和生长情况．行MTT比色、碱性磷酸酶（alkalin ephosphatase，ALP）活性定量检测、ALP和Von Kossa染色。结果：浓度为50ng／mL的bFGF可明显促进犬的BMSCs增殖（P〈0．05），但无显著促进犬BMSCs分化的作用（P〉0．051。结论：bFGF能有效促进犬BMSCs的增殖，且与bFGF剂量有关。     

bFGF对体外培养人牙囊细胞VEGF表达的影响

目的：研究碱性成纤维细胞生长因子（bFGF）对体外培养人牙囊细胞血管内皮生长因子（VEGF）表达的影响。方法：选取生长状态良好的人牙囊细胞，与10ng／mlbFGF孵育不同时间后，采用反转录聚合酶链反应（RT—PCR）及酶联免疫吸附试验（ELISA）分别检测人牙囊细胞VEGFmRNA表达及上清液中VEGF蛋白分泌变化。结果：bFGF作用1、3、6、12h后人牙囊细胞VEGFmRNA表达均较0h组显著增强（p〈0．01），其中bFGF作用6h后VEGFmRNA表达最强；细胞培养上清液中VEGF蛋白分泌量随时间延长逐渐增加，但bFGF处理组VEGF蛋白分泌量高于对照组（p〈0．01）。结论：bFGF能够促进体外培养人牙囊细胞VEGF的表达。     

上颌后牙区骨膨胀技术结合上颌窦闭式提升术植入CDIC种植体的探讨

目的：探讨一种创伤较小的后牙牙槽脊增宽，同期植入CDIC种植体的方法。方法：通过使用后牙骨挤压骨膨胀的技术获得初期固位。结果：两例病例获得成功，近期成功率达100％。结论：上颌后牙区骨挤压骨膨胀技术结合上颌窦闭式提升术植入CDIC种植体是一种解决后牙区骨萎缩的方法。