小干扰RNA抑制胃癌细胞肝素酶表达及侵袭的实验研究

目的 利用RNA干扰（RNAi）技术封闭胃癌细胞系SGC7901肝素酶表达,观察其对肿瘤细胞侵袭能力的影响。方法 体外转录合成小干扰RNA（siRNA）,经脂质体转染胃癌细胞系;结合逆转录（RT）-PCR,Western印迹检测类肝素酶mRNA及蛋白质表达;通过体外侵袭实验评价肝素酶RNAi后胃癌细胞系的侵袭能力变化。结果 肝素酶siRNA分子可以特异性地抑制SGC7901细胞肝素酶mRNA,抑制率达（70±6）％;肝素酶RNAi后SGC7901细胞的侵袭抑制率达（61±36）％。结论 靶向肝素酶基因的siRNA分子能特异性抑制其蛋白表达水平,降低肿瘤细胞的侵袭能力。肝素酶是促进胃癌细胞侵袭转移的关键分子,并可为抑制胃癌侵袭转移提供新策略。     

冠脉腔内支架置入术后联合抗凝的探讨

目的：评价冠脉腔内支架置术后低分子肝素、噻氯匹定和阿司匹联合抗凝的效果和安全性。方法：121例行经皮冠状动脉腔内支架置入术的患者，分成择期组86例，不稳定心绞痛低分于肝素抗凝不佳需行急仍支架入术者为急诊组1共15例，急性心肌梗死20例直接支架入术为急诊组2。急诊组1术前不停用分子肝素，术后低分子肝素0．4ml，每12h1次共7d；噻氯匹定0．25g／次，每天两次和阿司匹林0．1g／d，顿服共1月联合抗凝。结果：三组术后1例出现急性、亚急性闭塞，无严重的出血并发症，一周内无心绞痛发作。结论：冠脉腔内支架置术后低分子肝素、噻氯区定和阿司匹林联合抗凝可预防急性、亚急性闭塞，未增加要后出血的发生，安全性好。     

肝素结合的类表皮生长因子促进大鼠减体积肝移植术后移植肝细胞的再生

目的 探讨肝素结合的类表皮生长因子（heparin binding epidermal growth factor—like growth factor，HB-EGF）对大鼠减体积肝移植术后肝细胞再生的促进作用。方法 采用改良“二袖套法”建立大鼠原位减体积肝移植模型，分为实验组和对照组，术后即刻经尾静脉分别给予HB-EGF（500μg／kg）和相同体积的生理盐水，2次／d。2组分别在术后第6h、2d、4d及7d随机挑取5只大鼠处死，称取移植物湿重，采血检测血清丙氨酸转氨酶（ALT）及白蛋白（Alb），流式细胞仪检测移植肝细胞的增殖活性，免疫组织化学法检测移植肝Ki-67的表达情况。结果术后第6h、2d和4d，实验组移植物湿重较对照组相应时相明显增加（P〈0．05）；术后第6h、2d、4d和7d，实验组血清ALT水平较对照组相应时相明显降低（P〈0．05），而第4和7d时Alb水平较对照组相应时相明显增高（P〈0．05）；术后第2和4d时，实验组的移植肝细胞增殖指数和Ki-67表达较对照组高（2d：P〈0．01；4d：P〈0．05）。结论 大鼠原位减体积肝移植术后使用HB-EGF能够明显促进移植肝细胞的再生。     

低分子量肝素治疗重症急性胰腺炎的多中心前瞻性临床研究

目的探讨低分子量肝素（LMWH）治疗重症急性胰腺炎（SAP）的效果和对胰外器官的保护作用。方法将265例SAP患者随机分为常规治疗组（C组，130例）和常规治疗＋LMWH治疗组（LT组，135例），对比两组的有关临床指标和治疗效果。结果加用LMWH治疗后与C组比较，LT组APACHE11评分明显降低（P〈0．05），肝、肾功能明显改善（P〈0．05、P〈0．01），血钙升高显著（P〈0．05），胰腺损害的CT评分显著降低（P〈0．05），器官衰竭发生率明显下降（P〈0．01），而器官衰竭治疗成功率明显提高（P〈0．05），中转手术率和病死率均明显降低（P〈0．01）。结论在常规治疗基础上加用LMWH能有效提高SAP的治疗效果，降低病死率。     

肝素抑制儿童白内障术后炎症反应的临床研究

目的：评价肝素液在先天性白内障人工晶体植入术中应用，对术后人工晶体前膜、后囊混浊的影响．方法：术毕前将浓度为 ２０Ｕ／ｍｌ肝素乳酸 Ｒｉｎｇｅｒ液冲洗前房，手术结束时应用乳酸 Ｒｉｎｇｅｒ液灌吸前房，将残留的肝素吸出，时照组用相同液灌吸前房．结果：对照组反应性色素膜炎、人工晶体表面颗粒状纤维蛋白沉积、纤维蛋白膜、术后三月后囊混浊等均高于实验组．结论：肝素对儿童白内障术后炎症反应，人工晶体前膜及后囊混浊有明显的抑制作用．     

两种抗凝剂对实验用小型猪血液生理指标的影响

目的比较分析两种抗凝剂对实验用小型猪血液生理指标的影响。方法将22只成年的实验用小型猪,分别用EDTA三钾和肝素锂抗凝采血后,用日本光电MEK-7222K血球分析仪测定血液生理指标。结果使用不同的抗凝剂,同一批样本的血液生理指标中单核细胞绝对值(MON)有差异(P0.05),平均红细胞体积(MCV)、平均红细胞血红蛋白浓度(MCHC)、血小板比积(PLT)、血小板平均体积(PCT)、血小板平均体积(MPV)、淋巴细胞绝对值(LYM)、嗜酸性粒细胞绝对值(EOS)、嗜碱性粒细胞绝对值(BAS)、淋巴细胞绝对值(LYM)%、单核细胞绝对值(MON)%、嗜酸性粒细胞百分率(EOS)%和嗜碱性粒细胞百分率(BAS)%这12个指标有显著差异(P0.01);EDTA三钾抗凝的血液生理指标中血小板比积(PLT)、中性粒细胞绝对值(NEUT)和中性粒细胞百分率(NEUT)%有性别差异(P0.05),MON、EOS、MON%和EOS%性别差异有显著性(P0.01);肝素锂抗凝的血液生理指标中血小板体积分布宽度(PDW)、中性粒细胞绝对值(NEUT)和LYM%有性别差异(P0.05),MON、EOS、MON%、NEUT%和EOS%性别差异有显著性(P0.01)。结论不同的抗凝剂对实验用小型猪血小板和白细胞分类计数参数影响显著,进行相关实验时应妥善选择血液抗凝方式。     

肝素对患感染性疾病的新生儿组织因子途径抑制物影响的观察

目的:观察肝素对患感染性疾病的新生儿组织因子途径抑制物(TFPI)的影响。方法:采用酶联免疫吸附试验(ELISA)法检测了32例患感染性疾病的新生儿在使用肝素前和使用肝素后30～45分钟、6小时血浆TFPI的变化,并与患儿对照组、正常对照组及文献进行比较。结果:患儿治疗组使用肝素后30～45分钟血浆TFPI水平明显增高(t=3.953,P<0.001),6小时血浆TFPI又几乎降至用药前水平(t=0.141,P<0.05)。结论:新生儿对肝素的反应与成人不同。     

Engineering strategies to control vascular endothelial growth factor stability and levels in a collagen matrix for angiogenesis: The role of heparin sodium salt and the PLGA-based microsphere approach

New vessel formation is the result of the complex orchestration of various elements, such as cells, signalling molecules and extracellular matrix (ECM). In order to establish the suitable conditions for an effective cell response, the influence of vascular endothelial growth factor (VEGF) complexation with heparin sodium salt (Hp) on its pro-angiogenic activity has been evaluated by an in vitro capillary-like tube formation assay. VEGF with or without Hp was embedded into collagen gels, and the activated matrices were characterized in terms of VEGF activity and release kinetics. Taking into account the crucial role of Hp in VEGF stability and activity, VEGF/Hp complex was then encapsulated into microspheres based on poly(lactide-co-glycolide) (PLGA), and microsphere properties, VEGF/Hp release kinetics and VEGF in vitro activity over time were evaluated. Integrated microsphere/collagen matrices were developed in order to provide a continuous release of active VEGF/Hp inside the matrix but also a VEGF gradient at the boundary, which is an essential condition for endothelial cell attraction and scaffold invasion. The results confirmed a strong influence of Hp on VEGF configuration and, consequently, on its activity, while the encapsulation of VEGF/Hp complex in PLGA-microspheres guaranteed a sustained release of active VEGF for more than 30 days. This paper confirms the importance of VEGF stability and signal presentation to cells for an effective proangiogenic activity and highlights how the combination of two stabilizing approaches, namely VEGF/Hp complexation and entrapment within PLGA-based microspheres, may be a very effective strategy to achieve this goal.     

Fondaparinux for intra and perioperative anticoagulation in patients with heparin-induced thrombocytopenia candidates for peripheral vascular surgery: Report of 4 cases

IntroductionIntra and perioperative anticoagulation in patients with heparin induced thrombocytopenia (HIT), candidates for peripheral vascular surgery remains a challenge, as the best alternative to heparin has not yet been established. We evaluated the off-label use of fondaparinux in four patients with HIT, undergoing peripheral vascular surgery procedures.Presentation of casesFour patients of whom 3 men of a mean age of 66 years, with proven heparin induced thrombocytopenia (HIT) underwent two axillo-femoral bypasses, one femoro-popliteal bypass and one resection of a splenic artery aneurysm under fondaparinux. No intra or perioperative bleeding or thrombosis of new onset was observed.DiscussionIn the absence of a valid alternative to heparin for intra and perioperative anticoagulation in HIT, several other anticoagulants can be used in an off-label setting. However, no general consensus exist on which should be the one of choice. In this small series fondaparinux appeared to be both safe and effective.ConclusionsThese preliminary results seem to justify the off-label use of fondaparinux for intra and perioperative anticoagulation in patients with HIT, candidates for peripheral vascular surgery interventions.