内皮抑素基因抗血管生成治疗子宫内膜异位症的实验研究

目的研究重组腺病毒介导的鼠内皮抑素(mEndostatin)基因对子宫内膜异位症的抑制作用。方法构建携带mEn-dostatin基因的重组腺病毒载体,建立人子宫内膜异位症的裸鼠模型,随机分为Ad-mEndo组、Ad-lacZ组和PBS对照组3组(n=10),观察3组用药前后异位病灶的形态、组织学及微血管密度(MVD)变化。结果治疗20d后,Ad-mEndo组异位病灶体积(34·3±11·2mm3)明显小于Ad-LacZ组(93·3±10·7mm3)和PBS对照组(90·4±18·7mm3,P<0·01),也小于用药前病灶体积(103·3±13·1mm3,P<0·01)。Ad-mEndo治疗的异位病灶MVD(9·6±11·6),明显低于Ad-LacZ组(15·8±10·4)和PBS对照组(15·8±18·6,P<0·01)。光镜下可见Ad-mEndo组异位子宫内膜明显减少,腺腔狭窄,细胞稀疏,呈萎缩状态,或可见腺上皮细胞空泡变性。TUNEL法检测Ad-mEndo组有(30·8%±12·0%)细胞出现凋亡,高于Ad-LacZ组(12·2%±7·1%)和PBS组(15·5%±9·5%,P<0·01)。结论所构建的mEndostatin重组腺病毒可有效地表达具有生物活性的内皮抑素,可以使子宫内膜异位症种植块的血管生成减少,抑制种植块生长,为抗血管生成子宫内膜异位症的临床应用奠定基础。     

血管内皮生长因子在子宫内膜异位症中的作用研究进展

子宫内膜异位症（endometriosis,EMs）是一种严重危害女性健康及生活质量的疾病,其发病率逐年上升,发病年龄也逐渐年轻化。关于子宫内膜异位症的发病机制,目前存在多种学说,其中Sampson的经血逆流种植学说最受广大学者认可。Sampson提出经期时子宫内膜腺上皮和间质细胞可随经血逆流,经输卵管进入盆腔,种植于盆腔并继续生长,形成盆腔内异症。     

克拉霉素抑制鸡胚绒毛尿囊膜血管生成的实验研究

目的 :评价克拉霉素对活体血管生成的影响。方法 :采用鸡胚绒毛尿囊膜方法检测 ,克拉霉素配制成 10～15 0mmol L的浓度梯度 ,观察其对血管生成抑制效果的量效关系 ,成纤维细胞生长因子、磷酸缓冲盐和环孢素分别用作阳性和阴性对照 ,Leica图像分析系统测量实验区域的血管面积。结果 :克拉霉素浓度 >30mmol L时对鸡胚绒毛尿囊膜的血管生成产生明显抑制作用 ,抑制效果与药物剂量明显负相关。结论 :克拉霉素对活体血管的生成具有抑制作用 ,在肿瘤和其他病理情况下的抗血管形成治疗中可能具有一定的应用价值。     

血管内皮生长因子受体KDR在子宫内膜异位症中的表达

 目的探讨血管内皮生长因子受体(VEGFR)胎肝激酶(KDR/flk-1)在子宫内膜异位症(内异症)患者中异位、在位子宫内膜及正常子宫内膜组织中的表达及意义.方法采用半定量RT-PCR(逆转录聚合酶链反应)和免疫组化(SABC法)方法,从基因和蛋白质水平检测44例内异症患者子宫内膜组织(卵巢子宫内膜囊肿32份)及30份在位内膜组织(研究组)中VEGFRKDR的阳性表达率.选择40例非内异症患者的正常子宫内膜组织作为对照组.结果两组多数异位及在位子宫内膜组织中均可检测KDR的表达,内异症组在位内膜和异位内膜KDR相对表达强度明显高于对照组,差异有统计学意义(P＜0.05);不同月经周期KDR的表达,在增殖期KDR相对表达强度高于分泌期,差异有统计学意义(P＜0.05).结论 VEGF受体KDR在内异症在位内膜和异位内膜中有明显表达,可能VEGF受体参与内异症血管生成,促使异位病灶远处转移种植、浸润;在位内膜增殖期KDR相对表达强度高于分泌期可能与卵巢激素相关.     

子宫内膜异位症肿瘤相关性发病机制研究进展

子宫内膜异位症（EM），是生育年龄妇女的多发病、常见病，在生育年龄妇女中发病率可达10％～15％，且呈明显上升趋势，严重地影响妇女的健康和生活质量。20世纪80年代后期以来，随着分子生物学及免疫学技术的发展，在传统的种植、直接扩散、体腔上皮化生与诱导、淋巴与血管转移等学说的基础上，关于异位症的基础与临床研究主要围绕免疫、激素、环境、遗传等方面进行，但子宫内膜异位症的发病机制仍不清楚。大量研究表明，虽然它是个良性疾病，而异位子宫内膜的黏附、侵袭和血管生成“三步曲”（即“3A”模式）及细胞无限生长和凋亡数量减少，     

子宫内膜异位症患者腹腔液血管内皮生长因子的检测

 目的 探讨VEGF在子宫内膜异位症发病中的意义,以及其作为临床诊断指标的可行性,为子宫内膜异位症的临床诊断和治疗提供科学依据和新思路.方法 用双抗夹心ELISA法测定各组腹腔液中VEGF水平.结果 (1)EMs患者腹腔液中VEGF浓度Ⅰ～Ⅱ期和Ⅲ～Ⅳ期均显著高于对照组(P<0.01,P<0.05);但Ⅰ～Ⅱ期与Ⅲ～Ⅳ期比较差异无统计学意义(P0.05),且VEGF浓度与r-AFS分期无相关性(r=-0.323,P0.05).(2)EMs患者腹腔液中VEGF浓度卵泡期较黄体期显著升高(P<0.05);对照组VEGF浓度在卵泡期和黄体期的差异无统计学意义(P0.05);卵泡期腹腔液VEGF浓度在EMs组和对照组间比较,差异有统计学意义(P<0.01,P<0.05),黄体期腹腔液VEGF浓度在EMS组和对照组间比较,差异有统计学意义(P<0.05).结论 VEGF与子宫内膜异位症的发生、发展有关,EMs患者腹腔液VEGF浓度可能受卵巢激素周期性调节.     

腹壁子宫内膜异位症的MRI特征分析

目的 探讨腹壁子宫内膜异位症(AWE)的MR影像特征.方法 选取经手术病理结果证实的AWE患者6例,行MRI检查分析MRI表现特征.结果 6例AWE病变均位于腹直肌旁,T1WI和T2WI均为等或略高信号,弥散加权成像均为不均匀略高信号,5例在T2WI序列呈现彗星尾征改变(83.3％),4例在T2WI脂肪抑制序列呈现...     

腹壁子宫内膜异位症

腹壁子宫内膜异位症(Abdominal wall endometriosis,AWEMS)多见于妇产科手术，尤其是剖宫产手术的切口部位．误诊者时有发生。Blanco等报道，术前仅1／3诊断正确。1992-03～2002-03我院行剖宫产者2038例，发生腹壁子宫内膜异位症6例，现报告如下。     

肾脏子宫内膜异位症一例

泌尿系子宫内膜异位症少见 ,起病隐匿 ,临床症状常不典型 ,不易得到确诊 ,而且可以引起尿路梗阻 ,导致肾功能丧失。子宫内膜异位至肾脏极为罕见 ,笔者经文献检索仅发现国外用法文报告1例[1] ,国内未见报道。我院发现 1例 ,经手术病理证实 ,影像资料齐全 ,现结合文献报道如下。患者女 ,42岁。因左腰部胀痛 2ｄ伴全程肉眼血尿来我院就诊。既往患者常有经期时左腰部胀痛 ,有时伴有腹部绞痛及血尿 10余年。 3年前当地医院Ｂ超发现左肾下极小囊性病变 ,追查发现该病变逐渐增大。本次体检 :左腰部叩击痛 ,全程肉眼血尿。腹部平片、静脉肾盂造影及…     

软骨血管生成抑制剂抑制血管生成的实验研究

软骨血管生成抑制剂抑制血管生成的实验研究２００４３３上海海烟煤医院研究所沈先荣，贾福星，于志洁，徐惠，陈杞中图法分类号Ｒ７３．３６软骨含有多种抗肿瘤有效成分，主要可分为两大类：血管生成抑制因子和肿瘤细胞抑制因子。软骨血管生成抑制因子在国外已有一些研究...     

检测血管生长因子作用的鸡胚绒毛尿囊膜技术

采用改良的鸡胚绒毛尿囊膜技术，探讨血管生长因子在肿瘤发生发展过程中的作用。方法是将肿瘤细胞培养上清液或某些细胞因子，置于孵育９ｄ鸡胚的ＣＡＭ膜上，继续孵育３ｄ后取膜，观察新生毛细血管的数量、长度和粗细，发现恶性程度高的肿瘤培养上清液促血管生长作用较强。     

质粒pUDKH刺激鸡胚绒膜尿囊膜血管生成效应的研究

目的 :探讨携带人肝细胞生长因子基因的质粒pUDKH对 13日龄鸡胚绒膜尿囊膜血管生成的刺激效应。方法 :以甲基纤维素为载体 ,将不同剂量的pUDKH(1.0 ,2 .0 ,5 .0 μg)或空白质粒pUDK(5 .0 μg)置入 13日龄鸡胚绒膜尿囊膜上 ,于孵化箱继续孵育 3d后 ,观察鸡胚绒膜尿囊膜上小血管的密度。结果 :应用质粒pUDKH各组小血管的密度均明显较对照组大 (P <0 .0 5 ) ,且有剂量_效应关系。结论 :质粒pUDKH可显著刺激鸡胚绒膜尿囊膜上小血管的形成 ,故认为其具有在体内应用于治疗缺血性疾病的潜能     

Fluorescence analysis of a tumor model in the chorioallantoic membrane used for the evaluation of different photosensitizers for photodynamic therapy

The development of a tumor in the chicken chorioallantoic membrane (CAM) enables a more individualized understanding of the dynamics of the photosensitizer (PS) interaction with the tumor blood vessels and cells. Photogem^® and 5-aminolevulinic acid (ALA), a protoporphyrin IX (PpIX) precursor, were used as PS and their red fluorescence enabled the monitoring of PS dynamic distribution in the vessels and in the tumor. With a tumor model in CAM and fluorescence assessment, the aim of this study was to evaluate the PDT parameters comparing different photosensitezers. In this model, the topical application was chosen as the best way of drug delivery and widefield fluorescence images were at every 30 min. The images were processed in a MATLAB^® routine for a semi-quantitative analysis of the red fluorescence. PpIX formation in the blood vessels and in the tumor region was observed after 3 h and 1.5 h, respectively, whereas Photogem^® was accumulated in the tumor region after 2 h. The illumination was performed by a diode laser with emission centered at 635 nm and irradiance of 80 mW/cm² for 10 min. After PDT irradiation, the photobleaching for both compounds was observed. Photogem^® showed a reduced photobleaching, however, both PS induced a destruction of the tumor mass and vascular network in the treated area.     

Effect of in ovo immobilization on development of chick hind‐limb articular cartilage: An evaluation using micro‐MRI measurement of delayed gadolinium uptake

To examine the effect of immobilization on the development of articular cartilage, we assessed glycosaminoglycan (GAG) content in the chick articular surface by delayed gadolinium‐enhanced MRI of cartilage (dGEMRIC). Chick embryos were paralyzed by decamethonium bromide (DMB) from day 10 to either day 13 or day 16. The GAG content of the chick knee was compared with that of nonparalyzed chick embryos. Histologic analysis was unable to quantify GAG content; however, dGEMRIC demonstrated that GAG content was higher in the femoral condyles of the nonparalyzed embryos on day 13, and on day 16 the GAG content was lower in both the femoral condyles and the tibial plateaus of the nonparalyzed embryos. These results suggest that paralysis delays embryonic hind‐limb development. Osteoblastic activity at the cartilage canal, as demonstrated by staining for alkaline phosphatase (ALP), was present only in the nonparalyzed chick embryos on day 16. The GAG content of the cartilage decreased when the cartilage canals began to form on day 16. The effect of immobilization on hind‐limb development was indicated by the differences in the GAG content of the cartilage anlage measured by dGEMRIC in the developing knee joint of paralyzed and nonparalyzed embryonic chicks. Magn Reson Med, 2006. © 2006 Wiley‐Liss, Inc.     

Feasibility of in ovo diffusion tractography in the chick embryo using a dual-cooling technique

Purpose:

To evaluate the capability of a dual‐cooling technique in suppressing motion artifact and to evaluate the feasibility of the noninvasive muscle fibers tracking using DTI during chick embryonic development.

Materials and Methods:

Fifteen eggs were divided into three groups of 5 eggs each (one group for each imaging sequence), and eggs were imaged every 48 h from incubation day 4; embryos were imaged in ovo using three sequences of varying duration (T1, T2, and DTI). For each sequence, three preprocessing methods were used: no‐cooling (NC), single‐cooling (SC), and dual‐cooling (DC). Two independent observers assessed images for motion artifact. The results of different preprocessing methods used for each sequence were compared by the χ² test. The Cohen kappa test was used to assess the interobserver variability.

Results:

For T1 imaging, motion artifact was adequately suppressed by both SC and DC methods (χ² test; P > 0.05). For T2 imaging, motion artifact was also sufficiently suppressed by both SC and DC methods (χ² test; P > 0.05) except incubation day 19 (χ² test; P < 0.001). For DTI, motion artifact was less with DC than SC after 8 days (χ² test; P < 0.05). Hindlimb muscle fibers of chick embryo could be serially evaluated with DTI from 8 days using dual‐cooling technique.

Conclusion:

The dual‐cooling technique enables DTI of chick embryo in ovo with minimal motion artifact, which permits muscle fiber tracking by DTI during chick embryonic development possible, and can improve the imaging quality of conventional MRI with long duration and those sensitive to motion. J. Magn. Reson. Imaging 2012;36:993–1001. © 2012 Wiley Periodicals, Inc.     

MRI of the basement membrane using charged nanoparticles as contrast agents

The integrity of the basement membrane is essential for tissue cellular growth and is often altered in disease. In this work a method for noninvasively detecting the structural integrity of the basement membrane, based on the delivery of cationic iron‐oxide nanoparticles, was developed. Cationic particles accumulate due to the highly negative charge of proteoglycans in the basement membrane. The kidney was used to test this technique because of its highly fenestrated endothelia and well‐established disease models to manipulate the basement membrane charge barrier. After systemic injection of cationic or native ferritin (CF or NF) in rats, ex vivo and in vivo MRI showed selective accumulation of CF, but not NF, causing a 60% reduction in signal intensity in cortex at the location of individual glomeruli. Immunofluorescence and electron microscopy demonstrated that this CF accumulation was localized to the glomerular basement membrane (GBM). In a model of GBM breakdown during focal and segmental glomerulosclerosis, MRI showed reduced single glomerular accumulation of CF, but a diffuse accumulation of CF in the kidney tubules caused by leakage of CF through the glomerulus. Cationic contrast agents can be used to target the basement membrane and detect the breakdown of the basement membrane in disease. Magn Reson Med 60:564–574, 2008. © 2008 Wiley‐Liss, Inc.     

脂质体介导内皮抑素基因治疗对裸鼠人子宫内膜异位症的作用

目的 探讨脂质体介导内皮抑素(ES)基因治疗裸鼠人子宫内膜异位症(EMs)的疗效及不良反应.方法 BALB/c雌性裸鼠40只,建立皮下EMs模型后将动物随机均分为4组(n=10):A组,病灶局部注射脂质体介导的pcDNA3.1(+)-hES 20μg;B组,肌内注射脂质体介导的pcDNA3.1(+)-hES 20μg;C组,病灶局部注射脂质体介导的pcDNA3.1(+)空质粒20μg;D组,病灶局部注射等量培养液.观察治疗21d后裸鼠皮下异位病灶生长情况,检测病灶内微血管密度(MVD)、血管内皮细胞生长因子(VEGF)蛋白及治疗后第3、7天时VEGF mRNA的表达情况,并计算治疗21d后内生殖器(子宫、卵巢)重量及与体重的比值.结果 A、B、C、D组病灶生长倍增时间分别是7.49、7.02、6.67、6.15d;A、B组注射ES基因后病灶生长抑制率分别为90.51%、43.05%.治疗21d后,A组MVD值(32土10/mm~2)较B组(56±14/mm~2)、C组(82±19/mm~2)、D组(82士19/mm~2)明显减少(P<0.05),后三组间差异无统计学意义,四组间VEGF表达水平差异无统计学意义(P>O.05).与C、D组比较,A组在治疗第3天VFGF mRNA明显下降,第7天显著回升,而B组变化不明显.A、B组的内生殖器与体重比值(分别为0.008 6±0.002 5、0.008 O士0.003 4)较C、D组(分别为0.011 6士0.014 0、0.012 0士0.023 0)明显下降(P<0.05).结论 20μg脂质体介导pcDNA3.1(+)-hES治疗裸鼠人EMs有效,但须注意其对子宫、卵巢的影响.     

AdKDR-CDglyTK融合基因系统对胰腺癌血管生成的抑制作用

目的研究重组腺病毒介导的KDR启动子驱动的CDglyTK融合双自杀基因系统(AdKDR-CDglyTK)对胰腺癌血管生成的抑制作用。方法将20只荷人胰腺癌裸鼠随机分为4组,每组5只。Ⅰ组:注射重组腺病毒AdKDR-CDglyTK与前药5氟胞嘧啶(5-FC)、更昔洛韦(GCV);Ⅱ组:仅注射前药5-FC、GCV;Ⅲ组:仅注射重组腺病毒AdKDR-CDglyTK;Ⅳ组:空白对照,不施加任何处理。采用重组腺病毒AdKDR-CDglyTK瘤体内多点注射,5-FC与GCV腹腔内注射的方法,观察各组治疗效果;微血管密度计数(MVD)分析该系统对胰腺癌血管形成的影响;RT-PCR检测各组的肿瘤组织,以了解有无双自杀基因CDglyTK(目的基因)的表达。结果Ⅰ组裸鼠移植瘤的生长显著受到抑制,Ⅱ、Ⅲ、Ⅳ组肿瘤生长情况无明显差别。MVDⅠ、Ⅱ、Ⅲ、Ⅳ组分别为2.08±0.79,10.01±0.77,9.91±0.63,10.39±1.35,组间差异有统计学意义(F=93.29,P=0.00),Ⅱ、Ⅲ、Ⅳ组与Ⅰ组比较,差异有统计学意义(P<0.05),而Ⅱ、Ⅲ、Ⅳ组之间差异无统计学意义(P>0.05)。RT-PCR结果显示,Ⅰ、Ⅲ组的瘤组织内扩增出2.4kB的目的基因片段,而Ⅱ、Ⅳ组瘤组织内未扩增出目的基因片段。结论AdKDR-CDglyTK联合前药5-FC及GCV在体内可明显抑制裸鼠胰腺癌血管形成和肿瘤生长。