视网膜特异性表达人血管内皮生长因子 基因载体的构建

目的构建在视网膜组织特异性表达的人血管内皮生长因子（VEGF）₁₆₅基因。方法用聚合酶链反应(PCR)方法从BLAB/C鼠全基因组扩增能在视网膜组织特异性表达的rho启动子，经限制性内切酶纯化后克隆于质粒pcDNA^3.1+-VEGF₁₆₅中，建立重组质粒pcDNA^3.1+-rho-VEGF₁₆₅，通过限制性内切酶酶切分析及PCR鉴定筛选出正确重组质粒pcDNA^3.1+-rho-VEGF₁₆₅,由jetPEI介导转染人视网膜色素上皮细胞和人脐静脉内皮细胞，并通过免疫组织化学染色以及绘制细胞生长曲线检测在人视网膜色素上皮细胞和人脐静脉内皮细胞中VEGF蛋白的表达。结果在人视网膜色素上皮细胞中，重组质粒pcDNA^3.1+-rho-VEGF₁₆₅比质粒pcDNA^3.1+-rho-VEGF₁₆₅的VEGF蛋白表达强，在人脐静脉内皮细胞，两者的表达量无明显差别。结论pcDNA^3.1+-rho-VEGF₁₆₅载体的构建为进一步研究VEGF在视网膜新生血管形成中的致病机理提供基础材料，并为进一步建立视网膜特异性表达VEGF转基因鼠模型建立了基础。(中华眼底病杂志,2005,21:106-109)     

Effects of chlorpromazine as a systemic vasodilator during cardiopulmonary bypass in neonates

OBJECTIVES: Vasodilator use during cardiopulmonary bypass is important in pediatric cardiac surgery, but the full range of their effects on hemodynamics remains to be clarified. We studied the effects of chlorpromazine, a potent alpha-blocking agent, in neonates. METHODS: Subjects were 60 neonates undergoing arterial switch operations for complete transposition of the great arteries with an intact ventricular septum. Of these, 37 received 2.1 to 6.5 mg/kg of chlorpromazine during cardiopulmonary bypass (CPZ group) and 23 received no vasodilator (control group). We then compared hemodynamic parameters between groups during and early after surgery. RESULTS: The systemic vascular resistance index and mean arterial pressure during cardiopulmonary bypass were significantly lower in the CPZ group (p < 0.05), but systolic pressure 15 minutes after cessation of cardiopulmonary bypass did not differ between groups. The rise in peripheral temperature during rewarming after hypothermia was significantly higher and the acid-base status 40 minutes after cardiopulmonary bypass less acidotic in the CPZ group. Urine output during cardiopulmonary bypass was higher in the CPZ group. CONCLUSIONS: Chlorpromazine effectively counteracts systemic vasoconstriction induced by cardiopulmonary bypass without serious side effects in neonatal cardiac surgery.     

Pulmonary vascular disease in secundum atrial septal defect in childhood: an unusual evolution

Reported is a case of a 16-month-old girl with an isolated atrial septal defect in whom severe pulmonary hypertension has developed in 26 months in spite of an important functional gradient across the pulmonic valves at a first catheterization. Individual susceptibility to an increased pulmonary blood flow is evoked.     

脑血管血液动力学参数脑卒中预测模型的建立

目的 根据脑血管血液动力学参数(CVHI)和脑卒中的主要危险因素建立脑卒中预测模型。方法 选择全国六大行政区脑卒中研究队列人群25355例，将基线调查时的CVHI检测结果 进行主成分分析，再以各主成分和主要脑卒中危险因素为自变量，以随访中脑卒中发病为应变量进行回归分析，根据回归系数建立脑卒中预测模型，计算发病概率，绘制ROC曲线，确定最佳截断点，评价预测模型的预测效能。结果 四个主成分的累积贡献率依次为58．1％、79．4％、88．4％和94．6％，被筛检进入logistic回归方程的变量分别为第一至第四主成分、高血压病史、年龄和性别，ROC曲线下面积为0．855，最佳截断点为预测概率≥0．05，预测脑卒中的敏感度、特异度和准确度分别为踟．7％，78．5％，78．5％。结论 通过主成分回归分析，可以建立具有良好效能的脑卒中预测模型。     

人血管内皮生长因子C基因真核表达载体的构建

目的：构建人血管内皮生长因子C(vascular endothelial growth factor C，VEGF-C)基因的真核表达载体，以便进一步研究VEGF-C基因在淋巴管生成中的作用。方法：根据人VEGF-C的cDNA序列，设计合成一对5′端分别含有EcoR I和BamH I酶切位点的特异性引物，运用RT-PCR方法克隆人乳腺癌细胞MDA-MB-231中的VEGF-C cDNA(1．26Kb；回收PCR产物(1．28Kb），并将其连接至克隆载体pMDl8-T中；重组的pMDl8-T在大肠杆菌DH5α内扩增后，经质粒提取、EcoR I和BamH I酶切，筛选出阳性重组子，并进行基因测序鉴定；琼脂糖凝胶电泳回收含有VEGF-C cDNA全长的酶切片断(1．27Kb)，然后在DNA连接酶作用下将其与真核表达载体pcDNA3．1(-)连接，重组质粒经EcoR I和BamH I酶切予以鉴定。结果：RT-PCR产物合有VEGF-C cDNA，基因测序显示重组的pMDl8-T中含有正确的人VEGF-C cDNA全长序列，重组的pcDNA3．1（-)中含有人VEGF-C cDNA全长序列。结论：成功构建了人类VEGF-C真核表达载体VEGF-C／pcDNA3．1（-)。     

脂多糖和同型半胱氨酸对人脐静脉内皮细胞单核细胞趋化蛋白1 mRNA表达的影响

目的探讨脂多糖和同型半胱氨酸是否诱导培养的人脐静脉内皮细胞表达单核细胞趋化蛋白1(MCP-1) mRNA及其机制.方法将生长至汇合的人脐静脉内皮细胞分为对照组、脂多糖组,脂多糖+SB203580组,同型半胱氨酸组,同型半胱氨酸+SB203580组,脂多糖+同型半胱氨酸组.采用斑点杂交和RT-PCR检测其MCP-1 mRNA的表达.用免疫细胞化学法检测对照组、脂多糖组和同型半胱氨酸组P38蛋白激酶蛋白的表达.结果培养的人脐静脉内皮细胞能表达较低水平的MCP-1 mRNA.斑点杂交和RT-PCR均显示各实验组的MCP-1 mRNA明显高于对照组.免疫细胞化学显示,P38蛋白激酶蛋白在对照组的细胞核阳性表达率为9.6%,当人脐静脉内皮细胞暴露于脂多糖或同型半胱氨酸后,细胞核阳性表达率升至46.7%或57.7%.结论脂多糖和同型半胱氨酸能诱导人脐静脉内皮细胞表达单核细胞趋化蛋白1 mRNA,P38蛋白激酶可能参与了该过程.     

Different effects of halothane, isoflurane and sevoflurane on sarcoplasmic reticulum of vascular smooth muscle in dog mesenteric artery

Background: The direct effect of halothane on vascular smooth muscle is mediated in part via its effects on the sarcoplasmic reticulum (SR). Little information is available concerning the effects of other volatile anesthetics including isoflurane and sevoflurane, whose vascular effects differ from those of halothane. The aim of the present study was to compare the effects of halothane, isoflurane and sevoflurane on the SR by testing the contraction induced by caffeine in vascular smooth muscle. Methods: Rings without endothelium from isolated canine mesenteric artery were mounted in physiological saline solution (PSS) for isometric tension recording. After complete depletion of Ca²⁺ from the SR by adding 35 mM caffeine, the rings were exposed to normal Ca²⁺ containing PSS (Ca²⁺ loading), to Ca²⁺-free PSS for 10 min, and then to 15 mM caffeine to induce contraction. Anesthetics were administered during Ca²⁺ loading, the Ca²⁺-free phase and simultaneously with caffeine administration. Results: Halothane (0.5-2%) attenuated the caffeine-induced contraction of canine mesenteric artery when administered during Ca²⁺ loading in the SR (P<0.001), whereas isoflurane and sevoflurane (1–4%) failed to affect the contraction. When given simultaneously with caffeine, halothane (1–2%) potentiated the caffeine-induced contraction (P<0.05), but isoflurane and sevoflurane had no effect. When given before caffeine administration, halothane (0.5-2%), isoflurane (24%) and sevoflurane (4%) all potentiated the caffeine-induced contraction (P<0.05). Conclusion: It has been shown that halothane not only potentiates caffeine- induced Ca²⁺ release from the SR, but also induces contraction by releasing Ca²⁺ from the SR. We conclude that halothane decreases Ca²⁺ accumulation in the SR while exerting facilitative and additive effects on caffeine-induced Ca²⁺ release from the SR when applied before caffeine administration and simultaneously with caffeine, respectively, whereas isoflurane and sevoflurane lack both the ability to decrease Ca²⁺ accumulation and an additive effect on caffeine-induced Ca²⁺ release from the SR, but are able to facilitate Ca²⁺ release by caffeine.     

Changes in the microcirculation in rats with experimental trigeminal neuralgia

Laboratory of Pathophysiology of Pain and Laboratory of General Pathology of the Microcirculation, Research Institute of General Pathology and Pathological Physiology, Academy of Medical Sciences of the USSR, Moscow. Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 111, No. 1, pp. 9–11, January, 1991.