食管组织VEGF、HIF-1α、iNOS及ecNOS的表达导致食管静脉曲张

为探讨食管下段组织中血管内皮生长因子(VEGF)、低氧诱导因子-1α(HIF-1α)、诱导型一氧化氮合酶(i NOS)及内皮型一氧化氮合酶(ec NOS)的表达与门静脉高压症下发生食管静脉曲张之间的内在关联,本研究选取Wistar大鼠90只,以门静脉一步结扎法及结扎左肾上腺静脉行食管静脉曲张造模作为观察组,并以仅行门静脉主干分离,显露左肾、左肾静脉操作后的大鼠为对照组进行试验观察。研究发现,建模后第7、14、28天,观察组大鼠门静脉压力均显著升高(p0.05);在14、28 d时,观察组大鼠的HIF-1α、VEGF表达强度均明显高于对照组(p0.05),且建模后各时间点i NOS、ec NOS表达也均明显高于对照组(p0.05)。这表明VEGF、HIF-1α、i NOS及ec NOS的异常高表达,在食管静脉曲张的形成过程中发挥了重要作用,可为临床深入研究其发病机制提供理论依据。     

S-甲基异硫脲对大鼠门脉高压症食管静脉曲张的影响

目的观察诱导型一氧化氮合酶抑制剂SMT对大鼠门脉高压症食管静脉曲张的影响。方法健康雄性SD大鼠60只随机分为5组,假手术组、模型组、低剂量组、中剂量组和高剂量组。假手术组仅分离门静脉、左肾上腺静脉关腹,其余组门脉缩窄两步法加左肾上腺静脉结扎,建立门脉高压症食管静脉曲张模型。假手术组与模型组手术后给予腹腔注射生理盐水,其余3组手术后给予腹腔注射不同浓度SMT。手术后21 d,检测大鼠门脉血中TNOS、iNOS的活性及NO的浓度,免疫组化CD34标记食管血管内皮,测量每组大鼠食管横切面黏膜下血管的数目、面积。结果模型组大鼠门脉血中TNOS活性与iNOS活性以及NO浓度和食管黏膜下血管数目,血管平均截面积,血管总面积均较假手术组显著升高（P〈0.01）。中、高剂量组大鼠门脉血中TNOS活性与iNOS活性以及NO浓度和食管黏膜下血管的数目、血管平均面积、血管总面积较模型组均显著下调（P≤0.01）。结论大鼠门脉高压食管静脉曲张的发病机制中有NO参与,门脉缩窄型门脉高压食管静脉曲张病中NO主要由iNOS生成,SMT对大鼠门脉高压食管静脉曲张可能具有一定保护作用。     

多层螺旋CT门静脉成像在评估肝硬化门脉侧枝血管中的价值

目的：评价多层螺旋CT门静脉成像显示门静脉高压的价值。材料与方法：30例门脉高压患者进行了螺旋CT门脉成像检查,其中10例患者又进行了门静脉造影检查(间接法)。30例患者中全部存在侧枝循环,多数病例有2个或2个以上部位侧枝循环。结果：多层螺旋CT门静脉成像不仅显示了肝内门静脉2-3级分支,还显示了整套门脉侧枝血管系统。在三维门脉像上,脾门静脉曲张29例(占96．7％),其中1例脾静脉因栓子部分闭塞而狭窄,另有1例则完全栓塞血管未显示。胃左静脉曲张28例(占93％),食管或食管旁静脉曲张27例(占90％),胃短静脉(胃后静脉)曲张19例(占63％),胃肾分流血管10例(占33％),腹膜后静脉曲张9例(占30％),脐周静脉曲张伴腹壁静脉曲张6例(占20％)。10例患者CT三维门脉像与间接门静脉造影作比较,前者对门静脉及其侧枝循环的显示好于后者。结论：多层螺旋CT门脉成像是门静脉无创性检查的可靠方法,有较高的临床运用价值。     

氩激光光凝建立兔视网膜静脉阻塞模型及其评价

目的制备视网膜中央静脉阻塞的动物模型,为视网膜中央静脉阻塞疾病的研究提供稳定的模型。方法采用氩激光直接光凝法封闭兔眼视网膜静脉建立视网膜静脉阻塞模型,利用HE染色法观察正常兔及造模后3周兔视网膜组织中视网膜新生血管的增生情况,利用免疫组化法观察正常兔及造模后3周兔视网膜组织中血管内皮生长因子(vascular endothelial growth factor,VEGF)的表达,并检测造模前及造模后3周闪光视网膜电流图(flash electroretinogram,FERG)。结果兔视网膜静脉阻塞(retinal vein occlusion,RVO)眼可见明显的视网膜新生血管增生,缺血视网膜及新生血管组织中VEGF不同程度表达增强,FERG-b波的振幅明显下降(P<0.01)。结论采用氩激光直接光凝法建立视网膜静脉阻塞模型是成功的。     

梅花鹿肝内门静脉系统的初步观察

以铸型技术观察了梅花鹿（Ｃｅｒｖｕｓ　ｎｉｐｐｏｎ）肝的门静脉系统。发现门静脉左支发出左外侧叶背侧静脉、左外侧叶腹侧静脉、左内侧叶外侧静脉、左内侧叶内侧静脉,尾状叶支和方叶支;右支缺如;右内侧叶静脉、右外侧叶背侧静脉、右外侧叶腹侧静脉直接由门静脉分出,尾状突静脉由右外侧叶腹侧静脉分出。梅花鹿肝如人、兔、猪、山羊、猕猴一样可分为二叶、四段,即左、右叶和左外侧叶（段）、左内侧叶（段）,右内侧叶（段）、右外侧叶（段）,尾状叶的左、右部可分别隶属于左、右叶。     

食管胃底静脉曲张出血药物治疗进展

在慢性肝病晚期患者中,门静脉高压属于普遍存在的并发痘之一。临床上门静脉高压往往会引发食管胃底静脉曲张,最终出现破裂出血。随着研究的进一步深入,目前研究证实肝脏结构出现改变、肌纤维母细胞发生收缩以及缩血管活性物质分泌的增加是引发门脉阻力上升最主要的三个方面。研究表明,门脉阻力上升能够被一些药物所阻断,这为临床使用一些缩血管药物降低门静脉高压,预防食管胃底静脉曲张出血（EVB）提供了理论基础。     

肝硬化患者肝脏门静脉血流与肝叶萎缩之间关系的研究

目的:探讨肝硬化患者肝脏右叶、左叶体积变化,检测肝硬化患者门静脉血流情况,分析二者之间的关系,以及门静脉血流与肝功能之间关系。方法:本研究纳入54例肝硬化患者和40例正常人,采用超声多普勒方法分析这些受试者的肝脏体积和门静脉主干及左右分支的内径、血流速、流量数据,并通过静脉血检测白蛋白、胆红素、胆碱酯酶水平等评估患者肝功能水平。结果:肝硬化组平均年龄46.3岁,男性32例,其中child A级患者16例,child B级患者27例,child C级患者11例;正常对照组平均年龄41.8岁,男性24例。肝硬化组患者右左肝叶之比明显低于正常对照组(p<0.05),门静脉内径和血流量明显高于正常对照组(p<0.05).随着child分级升高,门静脉血流量也明显升高。肝硬化组门静脉右支血流量明显低于左支血流量(p<0.05);此外肝硬化患者门静脉右支和左支血流量之比明显低于正常人群门静脉右左支之比(p<0.05);而且肝硬化患者门静脉右左支血流量之比与右左肝叶具有明显的相关性与右左肝叶之比具有明显的相关性(r=0.64,p<0.05)。结论:评估肝硬化病人门静脉血流情况,对于判断肝脏病理变化程度,评价治疗效果,以及选择治疗方案方面都具有重要的临床价值     

肝硬化患者肝脏门静脉血流与肝叶萎缩之间关系的研究

目的：探讨肝硬化患者肝脏右叶、左叶体积变化,检测肝硬化患者门静脉血流情况,分析二者之间的关系,以及门静脉血流与肝功能之间关系。方法：本研究纳入54例肝硬化患者和40例正常人,采用超声多普勒方法分析这些受试者的肝脏体积和门静脉主干及左右分支的内径、血流速、流量数据,并通过静脉血检测白蛋白、胆红素、胆碱酯酶水平等评估患者肝功能水平。结果：肝硬化组平均年龄46.3岁,男性32例,其中childA级患者16例,childB级患者27例,childC级患者11例;正常对照组平均年龄41．8岁,男性24例。肝硬化组患者右左肝叶之比明显低于正常对照组（p〈0．05）,门静脉内径和血流量明显高于正常对照组（p〈0．05）．随着child分级升高,门静脉血流量也明显升高。肝硬化组门静脉右支血流量明显低于左支血流量（p〈0．05）;此外肝硬化患者门静脉右支和左支血流量之比明显低于正常人群门静脉右左支之比（p〈0．05）;而且肝硬化患者门静脉右左支血流量之比与右左肝叶具有明显的相关性与右左肝叶之比具有明显的相关性（r=0．64,p〈0．05）。结论：评估肝硬化病人门静脉血流情况,对于判断肝脏病理变化程度,评价治疗效果,以及选择治疗方案方面都具有重要的临床价值     

Caspase-3在心衰兔模型细胞凋亡过程中的变化

目的研究caspase-3在导管术致超容量负荷型心衰兔模型细胞凋亡中的变化。方法运用导管术造成兔的超容量负荷型心衰模型,然后分别于造模后24 h和造模后7 d处死兔,取心脏组织做电镜检查;通过流式细胞仪检测心肌细胞凋亡情况、caspase-3试剂盒检测其活性、Western blot法检测caspase-3蛋白水平的表达。结果①电镜检查表明：正常组心内膜完整无损伤,有少量红细胞附着;造模24 h后和造模7 d后均出现大量红细胞附着和血栓,且造模7 d的血栓较造模24 h减少。②AnnexinV联合PI染色结果表明：正常组凋亡率较低（3.72±1.92）%,随着时间推移,凋亡细胞所占比例呈上升趋势,造模7 d（10.73±2.27）%明显高于造模24 h（7.62±1.70）%。③Caspase-3活力测定：随时间推移,caspase-3活力呈上升趋势,造模7 d明显高于造模24 h,指数分别为2.7±0.31和2.15±0.32。④Western Blot检测结果表明：正常组基本检测不到激活的caspase-3蛋白酶,在造模后24h和造模后7 d caspase-3蛋白酶均有明显表达,但造模7 d有所下降。结论超容量负荷型心衰兔模型的细胞凋亡与caspase-3有密切相关。     

内镜套扎治疗门静脉海绵样变性食道静脉曲张破裂出血的疗效分析

目的:探讨内镜下曲张静脉套扎术(EVE)治疗门静脉海绵样变性引起食道静脉曲张破裂出血的疗效.方法:回顾性分析了2004年3月～2007年8月,我院采用内镜下曲张静脉套扎术治疗15例门静脉海绵样变性引起食道静脉曲张破裂出血的临床资料.结果:15例患者经EVL治疗均能有效控制出血,其急诊止血率达93.3%;经1-3次EVL治疗后曲张静脉基本消失,消失率达86.7%;随访6个月～4.1年,一年期静脉曲张复发率为38.5%,两年期静脉曲张复发率为33.3%,三年期静脉曲张复发率为25%,四年期静脉曲张复发率为0.在整个随访期中有8例患者自第一次EVL术后食管曲张静脉基本消失从未复发.结论:EVL治疗门静脉海绵样变性引起食道静脉曲张破裂出血是一种安全有效、简单易行的方法,可以起到急诊止血、消除曲张静脉和预防再出血的作用.     

束状刺盘孢角膜炎的实验研究

目的初步研究束状刺盘孢感染兔角膜的临床表现和组织病理学特征。方法选择15只新西兰白兔,应用基质内直接注射法建立模型。肉眼观察角膜变化,并在接种后第5 d、14 d、21 d处死家兔,对角膜行HE染色和PAS染色,观察其组织病理学改变。结果兔束状刺盘孢角膜炎动物模型成功建立,肉眼可以见到典型的真菌性角膜炎的表现。组织病理学检查,在感染早期即可看到大量真菌病原体生长,菌丝垂直于角膜基质,后期出现新生血管和瘢痕。结论角膜基质内直接注射法复制真菌性角膜炎动物模型成功率高、稳定。兔束状刺盘孢角膜炎有典型的真菌性角膜炎的表现,组织学中菌丝垂直于角膜基质生长。     

四氯化碳复合法诱导家兔肝纤维化模型的建立

目的建立一组家兔肝纤维化动物模型,并总结建模过程中所得到的经验。方法选用30只普通级家兔,随机选取2～4只家兔作为实验组,5只为对照组。饲养适应1周后,四氯化碳色拉油溶液以5%的起始浓度经腹腔注射法注入实验组家兔体内,对照组给予相同剂量的生理盐水。于造模4W、7W、10W、13W、16W末分别处死24只实验组及1只对照组家兔,以观察肝纤维化的发展程度。结果造模期间实验组家兔死亡7只,对照组家兔无死亡。家兔肝脏纤维化程度随着造模时间延长而逐渐加重。结论此方法成功地建立了一种兔肝纤维化动物模型。     

Electric and mechanical activities of the upper esophageal sphincter in rabbits

Electrical and mechanical activities of the rabbit muscles in different zones of the esophageal cervical part were examined on free-moving rabbits with chronically implanted electrodes and force transducers under conditions of hunger and food intake. It is shown that the functional role of the circular muscles of the cranial end of the esophagus is determined by their participation in activity of the superior esophageal sphincter.     

Bio-Plex悬浮芯片系统检测两种实验兔白介素基因的差异表达

目的采用液相悬浮芯片系统同时测定实验兔圆小囊中IL-1β、IL-1R1、IL=8、IL-8RA和IL=15各基因的表达情况,并对该方法进行评价。方法利用Affymetrix的Panomics QuantiGene Plex2．0Assay中bDNA信号放大和多磁珠分析技术,来同时检测两种实验兔圆小囊中多重mRNA并定量。建立实验兔免疫相关白介素基因的液相悬浮芯片检测方法。结果可同时检测IL-1β、IL=1R1、IL-8、IL=8RA和IL-15各基因的含量,并发现WHBE兔IL-15基因的相对表达量显著高于JW兔（P〈0．05）,IL-1R1基因的相对表达量显著高于JW兔（P〈0．01）,IL-8RA基因在WHBE兔中的相对表达量也高于JW兔（P〈0．05）。结论建立了实验兔白介素基因的液相悬浮芯片检测方法,WHBE兔的IL-15、IL-1R1和IL-8RA基因表达量较高,可能与WHBE兔独特的免疫学特性有关。     

A Canine Model for IVC Occlusive Form of Budd–Chiari Syndrome Using Endovascular Technique

The objective of this study was to assess the portal/hepatic changes in a newly designed canine model for Budd–Chiari syndrome (BCS). The inferior vena cava (IVC) was occluded using a diagram stent under general anesthesia in 10 mongrel dogs under the guidance of percutaneous angiography. Five dogs that received IVC angiography only were used as sham controls. Occlusion of the IVC increased the diameter of the hepatic veins, portal vein, and IVC. Massive ascites, significantly increased abdominal circumference, varying degrees of esophageal varices, congestion, cirrhosis, and fibrosis of the liver were also noted. BCS could be readily established by placing a diaphragm–stent in the IVC via a percutaneous endovascular approach.     

Establishment of a rabbit model of superior vena cava obstruction.

OBJECTIVE: To explore a method of establishing a rabbit model of superior vena cava obstruction (SVCO) by injecting VX2 tumor cell suspension transcutaneously under ultrasound guidance. METHODS: A suspension of VX2 tumor cells was prepared under sterile conditions. Fifteen adult healthy New Zealand White rabbits were enrolled in the experiment. Under ultrasound guidance, about 0.1 ml of the living tumor cell suspension was transcutaneously injected in front of the anterior wall of the right superior vena cava (SVC). The lumen, wall, blood flow of SVCs and adjacent tissues were examined with gray-scale and color Doppler ultrasonography, every 3 days starting from the 9th day after injection. Meanwhile, CT scanning and digital subtraction angiography (DSA) were also performed. The rabbits were dissected immediately after death and tissue samples were collected for pathologic examination. RESULTS: Fourteen out of 15 rabbits developed tumors that were located close to SVCs and/or SVCs cavity, which was shown by ultrasonography. The diameters of the tumors were 80.7 +/- 4.3 mm. These tumors grew close to SVCs area and resulted in compression and infiltration of SVCs. CT scanning and DSA confirmed the establishment of the SVCO model. The achievement rate of the SVCO model was 93.3%. No rabbit died of complications. CONCLUSION: A method of establishing a rabbit SVCO model by injecting VX2 tumor cell suspension under ultrasonographic guidance was established successfully, and it proved to be simple, effective and repeatable. The imaging characteristics of this model are in good accordance with those of SVCO in patients.     

脾虚证IBS模型WHBE兔血浆环核苷酸水平的变化

目的观察脾虚型肠易激综合征（IBS）模型WHBE兔环核苷酸水平的变化。方法取WHBE兔和日本大耳白兔各18只,分别随机分成两组,即正常对照组6只,模型对照组12只,采用湿热应激加灌服番泻叶煎剂的方法造成脾虚证肠易激综合征（tBs）免模型,其中模型对照组造模后处理6只,剩余兔自然恢复10d,观察实验兔的血浆环核苷酸水平的变化。结果与正常对照组比较,WHBE兔模型对照组血浆cAMP和cGMP水平均显著升高（P〈0．05,P〈0．01）,且自然恢复10d后血浆cAMP和cGMP水平仍高于正常对照组（P〈O．05,P〈O．01）;日本大耳白兔模型对照组造模后及自然恢复10d后血浆cAMP和cGMP水平略有所升高,但差异均不显著（P〉O．05）;且模型对照组和自然恢复组cAMP／cGMP比值均略低于正常对照组,但差异不显著（P〉O．05）。结论本实验脾虚证IBS模型兔的血浆环核苷酸水平随阴阳盛衰而变化,具有双重特点：既有阴虚特点,又有阳虚特点,且WHBE兔更适合于脾虚证IBS模型研究理想的实验动物。     

Establishment of a cottontail rabbit papillomavirus/HLA-A2.1 transgenic rabbit model

Three transgenic rabbit lines that express a well-characterized human major histocompatibility complex class I (MHC-I) gene (HLA-A2.1) have been established. All three lines carry the HLA-A2.1 heavy chain and are able to pass the transgene to their offspring with both the outbred and the inbred EIII/JC genetic background. HLA-A2.1 colocalizes exclusively with rabbit MHC-I on the cell surfaces. These HLA-A2.1 transgenic rabbits demonstrated infection patterns similar to those found after cottontail rabbit papillomavirus (CRPV) challenge when compared with results in normal rabbits, although higher regression rates were found in HLA-A2.1 transgenic rabbits. Because the CRPV genome can accommodate significant modifications, the CRPV/HLA-A2.1 rabbit model has the potential to be used to screen HLA-A2.1-restricted immunogenic epitopes from human papillomaviruses in the context of in vivo papillomavirus infection.     

A novel canine model of portal vein stenosis plus thioacetamide administration-induced cirrhotic portal hypertension with hypersplenism

Current large animal models that could closely resemble the typical features of cirrhotic portal hypertension in human have not been well established. Thus, we aimed to develop and describe a reliable and reproducible canine cirrhosis model of portal hypertension. A total of 30 mongrel dogs were randomly divided into four groups: 1 (control; n = 5), 2 (portal vein stenosis [PVS]; n = 5], 3 (thioacetamide [TAA]; n = 5), and 4 (PVS plus TAA; n = 15). After 4-months modeling period, liver and spleen CT perfusion, abdominal CT scans, portal hemodynamics, gastroscopy, hepatic function, blood routine, the bone marrow, liver, and spleen histology were studied. The animals in group 2 (PVS) developed extrahepatic portosystemic collateral circulation, particularly esophageal varices, without hepatic cirrhosis and portal hypertension. Animals from group 3 (TAA) presented mild cirrhosis and portal hypertension without significant symptoms of esophageal varices and hypersplenism. In contrast, animals from group 4 (PVS + TAA) showed well-developed micronodular and macronodular cirrhosis, associated with significant portal hypertension and hypersplenism. The combination of PVS and TAA represents a novel, reliable, and reproducible canine cirrhosis model of portal hypertension, which is associated with the typical characteristics of portal hypertension, including hypersplenism.