目的 研究重组pCKM-mPTH质粒在SD大鼠甲状旁腺功能低下症模型中的治疗作用。方法 将30只SD大鼠随机分为3组，A组为正常组，肌肉注射生理盐水，B组、C组制作甲状旁腺功能低下症模型后，分别肌肉注射pcDNA3.1（+）空质粒和pCKM-mPTH质粒。用放免法测定不同时间的血清甲状旁腺激素（PTH）浓度。结果 动物模型在注射pCKM-mPTH质粒后，其血清PTH浓度在近1个月内显著高于注射前及空质粒对照组，未观察到有甲状旁腺功能低下症的表现。结论 应用pCKM-mPTH质粒治疗SD大鼠的甲状旁腺功能低下症是有效的，对未来的临床应用研究有一定的指导意义。

目的 总结内镜筋膜下交通静脉结扎术（subfascial endoscopic perforator surgery，SEPS）治疗重度慢性下肢静脉功能不全的临床经验。方法 从1999年11月至2002年8月采用SEPS治疗重度慢性下肢静脉功能不全51例，共64条肢体。采用临床分级、静脉功能评分、彩色多普勒超声、彩色多普勒剖面流速图、静脉造影、空气体积描记仪等方法在手术前后对患肢状况进行分析。结果 术后随访率90.6%（58/64），随访3～35个月。全组浅静脉曲张未见复发，有40条肢体溃疡愈合（97.6%），愈合时间平均（32±5）d，手术前后静脉功能综合评分均值分别为（13.9±1.7）分vs.（6.4±0.8）分、手术前后彩超剖面流速图静脉返流量均值分别为（61.2±7.7）ml/svs.（45.6±5.7）ml/s和空气体积描记仪3项指标术前后均值比较均差异有显著意义（P<0.01）。结论 SEPS对于重度慢性下肢静脉功能不全，特别是合并静脉性溃疡者具有良好疗效，如同时行浅静脉缝扎和深静脉瓣膜修复成形可取得更好的疗效。

目的 筛选和克隆原发性下肢深静脉瓣膜功能不全的大隐静脉曲张的致病相关基因。方法 利用mRNA荧光差异显示技术比较原发性下肢深静脉瓣膜功能不全患者的曲张大隐静脉隐-股瓣膜区组织中mRNA表达与对照组的差异，获得的差异表达cDNA片段经Northernblot验证后进行克隆和测序以探明其基因来源。结果 共获得37条差异表达的cDNA条带，经杂交验证其中30条为阳性（阳性率80%）。克隆及测序结果显示C610片段（NO.18克隆）与麦-考综合征（Mckusick-Kaufmansyndrome，MKKS）基因的mRNA序列有96%的同源性。结论 C610片段（NO.18克隆）与麦-考综合征基因高度同源，与下肢大隐静脉曲张的发病密切相关。

目的 探讨白细胞介素8（IL-8）在乳腺癌细胞中促血管生成作用，及其与雌激素受体之间的关系。方法 收集不同的乳腺癌细胞培养上清液，通过观察这些上清液对人脐静脉内皮细胞迁移、增殖和新生毛细血管管腔形成三个体外血管生成实验以及裸鼠体内血管生成实验来研究细胞因子在乳腺癌血管生成中的作用机制；采用细胞上清液芯片实验检测MDA-MB-231细胞和雌激素受体稳定转染的MDA-MB-231细胞的IL-8表达水平；进一步将雌激素受体和IL-8启动子瞬时转染至MDA-MB-231细胞，经双荧光素酶报告基因检测雌激素受体对IL-8的调控作用。结果 与IL-8低表达细胞相比较，IL-8高表达和中表达细胞上清液所培养的HUVEC更易于发生迁移（t值分别为6.94和17.75，P均<0.05），IL-8抗体可以特异性地阻断这种作用（t值分别为16.67和9.08，P均<0.05）。雌激素受体阴性MDA-MB-231细胞经过雌激素受体稳定转染后，IL-8的表达水平下降8.8倍。雌激素受体α能够下调IL-8启动子的活性（r=0.856，P<0.05），对照载体无此作用。结论 IL-8具有促乳腺癌细胞血管生成作用，乳腺癌细胞中IL-8的水平与雌激素受体水平呈负相关，外源性的雌激素受体可下调乳腺癌细胞中的IL-8表达。

Despite the availability of effective treatments for Graves' disease, there are patients who are unable to tolerate, or choose not to accept, current therapies (oral medication, radioactive iodine, or surgery). In this study, we have examined the use of thyroid arterial embolization as an option for patients who suffer from Graves' disease and who fit into one of the preceding patient groups. Selective arteriography, followed by embolization of thyroid arteries, was performed using Seldinger's technique in 22 patients with Graves' disease. Six of the patients received both arterial embolization and surgery at 2-3 wk after embolization. No serious complications were noted in any patient. In those who were treated with only interventional embolization, 14 of 22 (63.6%) became euthyroid and remained in this state for the duration of the study. The remaining 2 patients required a maintenance dose of antithyroid drug therapy (tapazole, 5-10 mg/d; or propylthiouracil, 50-100 mg/d). Patients were followed for a median time of 27 months (range, 6-50). At the end of the follow-up period, all 22 patients were euthyroid, the size of thyroid gland had decreased by one third to one half its original volume, but 2 patients continued to require antithyroid drugs. Histologic analysis of thyroid tissue from 6 patients who had embolization followed by surgery showed that embolization blocked both the superior and inferior thyroid arteries plus most of their branches. Evidence of chemical inflammation, necrosis, and fibrosis were observed in embolized thyroid tissue. The average diameter of capillary network within the body of the thyroid adjacent to superior and inferior artery was 0.12-0.25 mm, with the smallest ones ranging from 0.04-0.11 mm. The diameter of the capillaries in the isthmus ranged from 0.13-0.15 mm. The average diameters of the superior and inferior arteries were 2-5.5 and 3-3.75 mm, respectively, measured using radiographs from angiography. Based on the measured diameters of arteries, the appropriate size of embolizing granules can be selected, and complete embolization of the majority of the thyroid glands was achieved with a high frequency of therapeutic success. Histological examination of the thyroid tissue and clinical follow-up after embolization indicate that this procedure is an effective, minimally invasive, and safe method for the treatment of Graves' disease patients who cannot, or choose not to, accept current therapies.

Tumor progression largely depends on blood supply and neovessel formation, and angiogenesis is emerging as a promising target for cancer therapy. Vascular endothelial growth factor (VEGF), a major proangiogenic molecule, stimulates angiogenesis via promoting endothelial proliferation, urvival and migration. VEGF has been found to be up-regulated in various types of tumors and to be associated with tumor progression and poor prognosis. Inhibition of VEGF or its signaling pathway has been shown to suppress tumor angiogenesis and tumor growth. In the present study, we tested the antiangiogenic and antitumor effects of soluble VEGF receptor-1 [soluble Flt (sFlt)-1] on the growth of follicular thyroid carcinoma (FTC). We constructed a 293 embryonic kidney cell line (293-Flt1-3d) that expresses sFlt-1, which is composed of the first three extracellular domains of Flt-1. The 293-Flt1-3d cells inhibited the in vitro growth of human umbilical vein endothelial cells in a paracrine manner. The in vivo antitumor and antiangiogenic activities of the 293-Flt1-3d cells were tested. When 293-Flt1-3d cells were inoculated at a site remote to the FTC-133 tumor transplant, the growth of FTC-133 tumors were inhibited by 70.37%, as compared with the control treatment with 293 cells expressing control gene LacZ. Immunohistochemical analysis of microvessel densities in treated tumors demonstrated that 293-Flt1-3d cells robustly suppressed intratumoral angiogenesis. Our data suggest that a mammalian cell-mediated approach could effectively deliver sFlt-1 gene therapy and inhibit tumor angiogenesis and tumor growth.

目的 研究对前甲状腺激素原基因进行定点突变，并转染体细胞产生成熟甲状旁腺激素。方法 采用聚合酶链反应（PCR）定点突变技术，自组织基因组基因中扩增前甲状旁腺激素原的cDNA，并将野生型甲状旁腺激素编码第28位的密码子GTT突变为AGA，使N端28-31氨基酸变为Arg-Lys-Lys-Arg，成为furin酶的酶切位点；将获得的突变表达载体pcDNA3.1/mPTH通过脂质体法转染体外培养的293细胞，用放射免疫法测定表达水平。结果 成功地获得突变型甲状旁腺激素原cDNA，293细胞转染突变表达载体后，每日培养液中甲状旁腺激素含量达28.34～52.64pg/2.0×106细胞，远高于空载体转染的细胞培养液。结论 利用PCR技术可获得突变型甲状旁腺激素原的cDNA，突变表达载体pcDNA3.1/mPTH能成功转染体细胞并产生甲状旁腺激素。