Transplantation of VEGF165-gene-transfected endothelial progenitor cells

Background: To study the cotherapeutic effect of stem cell transplantation and gene transfection on chronic venous thrombosis. Methods: We constructed a recombinant adenoviral vector carrying the VEGF165 gene by using the pAdEasy system, which was subsequently identified and amplified. Simultaneously, endothelial progenitor cells (EPCs) were isolated from rat bone marrow by using Ficoll, cultured in EBM-2MV medium, and identified. Then, the cells were transfected with the recombinant Ad-VEGF165. The EPCs were labeled with 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine (Dil) before transplantation. An experimental rat model of chronic vein thrombosis was developed by partial ligation of the inferior vena cava. The rats were randomly divided into 4 groups: A (n = 25), Ad-VEGF165/EPC-transplantation group, which received 1 ml (106) of Ad-VEGF165/EPCs; B (n = 25), EPC-transplantation group, which received 1 ml (106) of EPCs; C (n = 25), Ad/EPC-transplantation group, which received 1 ml (106) of Ad/EPCs; D (n = 25), control group, which received 1 ml of the transplantation medium. The thrombi and adjacent caval walls were harvested 28 days after transplantation; real-time quantitative polymerase chain reaction (PCR) was used to detect the expression level of VEGF mRNA; and western blotting was used to measure thrombosis and changes in VEGF protein expression. Hematoxylin-eosin (HE) staining and immunohistochemical staining were performed to detect recanalization, and neovascularization was observed by scanning electron microscopy. The capillary density was quantitatively determined by counting the capillaries under a high-power microscope. SPSS 11.5 software used for analysis, and differences at P < 0.05 were considered to be significant. Results: The Ad-VEGF165 was constructed, and bone-marrow-derived EPCs were cultivated and successfully identified. We determined the optimum transfection ratio that promoted the growth of EPCs. After transfection, the EPCs secreted the VEGF protein. After transplantation, the in vivo survival of EPCs and their differentiation into endothelial cells were determined by detecting the fluorescence associated with the Dil stain. VEGF mRNA was expressed in groups A, B, C, and D after transplantation, and the VEGF mRNA level in group A was significantly higher than those in groups B, C, and D (P < 0.05); the VEGF mRNA levels in groups B and C were significantly higher than those in group D (P < 0.05), and there was no statistical significance between the VEGF mRNA levels in groups B and group C. The recanalization capillary density in group A was significantly higher than those in groups B, C (P < 0.05), and D (P < 0.01); the recanalization capillary densities in groups B and C were significantly higher than that in group D (P < 0.05). Moreover, there was no statistical significant difference between the values for groups B and C. Neovascularization was detected by immunohistochemical staining using the antibody for vWF, which is a component of endothelial cells. Conclusion: The EPCs were successfully transfected by Ad-VEGF165. A suitable transfection ratio can improve the efficiency of EPCs, improving the possibility of promotion of angiogenesis after transplantation. Transfected EPCs caused accelerated organization and recanalization of vein thrombi.     

改良锁骨下静脉穿刺与长期留管

深静脉穿刺国内留管时间最长为459d。本组留管最长者已大于3年4个月，且无感染，目前仍继续留用。现将我们的方法和体会报告如下。     

臀肌注射深度与硬结形成的临床分析

肌肉注射是临床常用的治疗手段,而臀肌注射又最为多用。本文的主要目的是论证注射进针深度与药物硬结形成的关系,为在临床工作中根据体形选择注射用针提供依据。一、方法临床观察:采用自然观察法,对临床上实施臀部肌注药物的病人随机取样观察。病人按照体型分组,即分为:体重(公斤)≤身高(厘米)-110;体重≥身高-100;体重>身高-100等三组,以间接推断皮下脂肪之多寡。观察皮下脂肪多寡与药物硬结形成之关系。尸体标本观察:采用经甲醛防腐处理过的成年尸体,用解剖法观察臀肌部位的皮下脂肪厚度,以佐证临床观察的研讨目的。二、结果临床观察组:共观察104名病人,均为女性青年,平均年龄为21岁。注射药物为:青霉素、链霉素和庆大霉素,注射时间3—15天(平均9     

下腔静脉滤器内血栓形成的腔内治疗

急性深静脉血栓(deep venous thrombosis,DVT)或慢性DVT并急性血栓形成可导致肺栓塞(pulmonary embolism,PE)等疾病.下腔静脉(inferior vena cava,IVC)滤器植入术可有效预防肺栓塞,但是少数患者术后会发生IVC滤器内血栓形成.分析2008年3月至2011年3月苏州大学附属第二医院收治下肢DVT行IVC滤器植入患者116例,其中滤器内血栓形成患者15例,经腔内治疗取得良好效果.     

miR-126对内皮祖细胞增殖和迁移的影响以及靶基因的检测

目的 研究miR-126(micro RNA-126)对大鼠骨髓源性内皮祖细胞(endothelial progenitor cells,EPCs)增殖和迁移能力的影响,并探讨miR-126新的靶基因.方法 采用电转的方法,在EPCs中转染对照寡核苷酸和miR-126的模拟物或抑制物.噻唑蓝(MTT)法检测细胞增殖,划痕和transwell实验检测细胞迁移能力的改变.microRNA靶基因预测软件TargentScan在线分析miR-126的靶基因,并进一步用Western blot检测靶基因的表达变化.结果 (1) miR-126模拟物在转染后24 h对EPCs的增殖有促进作用(P＜0.01),转染后48和72 h,miR-126对EPCs增殖没有影响.(2)划痕和transwell实验证实miR-126模拟物可以促进EPCs的迁移(P＜0.01),miR-126抑制物抑制EPCs的迁移(P＜0.01).(3)TargetScan在线软件预测KANK2是miR-126的靶基因.(4) Western blot检测结果显示miR-126模拟物抑制KANK2的表达,miR-126抑制物促进KANK2的表达.结论 miR-126对EPCs的增殖有一过性的促进作用,miR-126可以促进EPCs的迁移能力并靶向KANK2蛋白,抑制KANK2蛋白的表达.     

应用间接补体结合试验检测猪衣原体抗体的研究

用鹦鹉热衣原体标准株B11001制备的抗原,对人工感染猪血清进行直接补体结合试验(DCF)和间接补体结合试验(ICF)时发现,猪血清中的衣原体抗体只能被ICF检出。于感染后第五天就能检出血清中的特异性抗体,感染后14个月抗体滴度仍在1:8以上。应用ICF对28头实验感染猪血清检测,检出率为100％。对本省9个地区34个猪场的857头猪进行检测,256头猪血清中发现有诊断意义的衣原体抗体,检出率为29.87％。     

剖宫产术182例麻醉体会

自 2 0 0 0 - 0 1～ 12 ,我院在 182例剖宫产手术中实施硬膜外麻醉 (ＣＥＡ)和腰硬联合麻醉 (ＣＳＥＡ) ,两种麻醉方法分组进行对比观察 ,现报道如下。1　资料与方法1.1　一般情况　 182例剖宫产孕妇 ,年龄 2 4 - 35岁 ,平均年龄 2 4 .5岁 ,ＡＳＡⅠ～Ⅱ级 ,均无椎管内麻醉禁忌症 ,随机分两组 ,Ａ组 78例 ,占 4 2 9% ,选用ＣＥＡ ;Ｂ组 10 4例 ,占 57 1% ,选用ＣＳＥＡ。1.2　方法　两组常规术前准备 ,麻醉前先输入 30 0～ 50 0ｍｌ平衡液 ,取左侧卧位Ｌ2 - 3间隙穿刺 ,Ａ组硬膜外置管 ,选取 1.5%～ 2 %利多卡因阻滞麻醉 ;Ｂ组用联合穿刺针…     

创伤性外周动脉病变的腔内治疗

外周动脉损伤是临床急症,虽然传统手术仍是治疗动脉损伤的金标准,但腔内技术有着独特的优势.苏州大学附属第二医院2006年1月至2011年6月应用血管腔内技术治疗了11例创伤性外周动脉损伤,现报道如下. 临床资料 1.一般资料:本组11例,临床资料见表1.其中例7和例9为动脉硬化闭塞症,行动脉闭塞段球囊扩张时发生血管破裂;例10左侧腘动脉损伤病例,入院前4h因重物砸伤左膝关节致开放性膝关节脱位,外院造影示左腘动脉中断后转入我科.     

新洁尔灭治疗马蜂蜇伤42例

我院外科门诊自1984年以来将其1/1000水溶液湿敷治疗马蜂蜇伤42例,效果满意。蜇伤部位:头、面、颈部30例,胸、腹、背部7例,四肢5例;两处以上蜇伤5例,单处伤37例。蜇伤后距就诊时间:约10分钟至1小时,就诊时症状:局部剧痛42例,红肿硬结直径约1～5cm,4例有水泡形成者。全身反应:全身皮肤潮红有荨麻疹者10例;     

麻醉下推拿治疗粘连性肩周炎306例

本文用臂丛麻醉推拿剥离粘连性肩周炎360例全部成功,随访6个月至3年201例,7例复发,其中2例再次推拿成功。本文就麻醉方法、推拿步骤和体疗方法进行探讨,介绍简易的臂丛电测定定位法。