Expression of Adhesion Molecules and Cytokines on Saphenous Veins in Chronic Venous Insufficiency

The objective of this study was to assess the relationship of signaling molecules to monocyte/ macrophages as a precursor to venous valve and venous wall dysfunction in patients with varicose veins. One of the hallmarks of venous dysfunction is destruction of venous valves with subsequent reflux and elevation of distal venous pressure. We recently observed that monocytes/macrophages migrate into the venous walls and valves of patients with venous insufficiency. There, they may play a role in the pathogenesis of primary venous insufficiency. If so, an important element in their performance would be the interaction between the monocytes and the endothelium as a precursor of damage to venous valves and the venous wall. To explore this interaction, immunohistochemistry was carried out to detect adhesion molecules and cytokines in surgical specimens removed during surgical therapy. Twenty-four surgical specimens consisting of proximal saphenous vein and subterminal valve were obtained using minimally traumatic technique in 6 males and 18 females who ranged in age from 31 to 79 years. Reflux was confirmed preoperatively by duplex technique, and severity was classified by the CEAP classification of the American Venous Forum. Ten patient limbs were class 2, eight were class 3, four were class 4, and two were class 6. The venous specimens were labeled using monoclonal antibody against ICAM-1, E-selectin, IL-1alpha, and TNF-alpha. CD68 was used for detection of monocytes/macrophages. Our results indicate that not only luminal venous endothelium but also endothelium in the vasa vasora of refluxing saphenous veins is activated, as indicated by the up-regulation of ICAM-1. However, IL-1alpha and TNF-alpha were increased in only selected specimens and are mainly detected in the vein wall. The factors that serve as trigger mechanisms to activate cells in the pathogenesis of primary venous dysfunction remain to be explored.     

Monocyte infiltration into venous valves

Purpose: Greater saphenous veins removed at surgery for correction of venous dysfunction have few valves. Those that are present may be shrunken, deformed, or monocuspid. This study was carried out to determine whether leukocyte infiltration is associated with valve damage. Methods: Seventeen specimens were removed at surgery from five men and eight women (age, 29 to 80 years). These consisted of the proximal 15 cm of the duplex-confirmed, refluxing proximal greater saphenous vein, one proximal lesser saphenous vein, and one midportion of greater saphenous vein. The severity of venous stasis in each patient limb was classified by the CEAP formula.¹ Twelve were class 2, two were class 3, and three were class 4. Control specimens were obtained from patients who underwent coronary artery bypass grafting. These were two men and two women, ages 72, 66, 62, and 60 years, free of venous insufficiency, with the specimens obtained from the proximal saphenous vein. Two of the control specimens contained venous valves corresponding to the test specimens. Longitudinal 10 μm paraffin sections were labeled with anti-CD64 monoclonal antibody, specific for tissue monocytes and macrophages, and studied by light microscopy. Five regions were chosen for quantification of the leukocyte infiltrate. Cells were categorized and counted directly. Volume proportions were calculated using stereologic techniques. Results: Three of seven specimens studied for morphologic changes had clearly shortened valve leaflets. Collagen degeneration was noted in all seven specimens. Leaflets had essentially disappeared in three and were shortened to 100 to 2100 μm in five. Specific leukocyte staining was accomplished on 10 additional specimens. All specimens showed monocyte/macrophage infiltration in valve leaflets and venous wall. These were more numerous in the valve sinus and proximal wall both on and under the endothelium. Control specimens showed no monocyte/macrophage infiltration. Conclusions: These observations suggest that venous valve damage in refluxing saphenous veins is associated with a leukocyte (monocyte/macrophage) infiltrate. Cell activation and fluid dynamic factors, such as eddies recirculation, and stasis in the valve sinus may be a part of the process of leukocyte penetration of the endothelium. The magnitude of leukocyte infiltration in the vein wall and in the base of the valve leaflet may be important in the genesis of primary venous dysfunction. (J Vasc Surg 1998;27:158-66.)     

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目的 利用基因芯片技术分析原发性下肢深静脉瓣膜功能不全(PDVI)所致大隐静脉曲张的基因表达谱和差异基因。方法 对10例PDVI所致的曲张大隐静脉和10例正常对照组织进行mRNA抽提，分别给予荧光素Cy5和Cy3标记以制备cDNA探针，并与含有1220个人类全长互补DNA的基因芯片进行杂交和分析。结果 在PVDI所致的曲张大隐静脉组织中共检测到21个差异基因，其中9个为上调基因，12个为下调基因。差异表达基因包括参与信号传导、细胞周期以及细胞生长、分化、代谢、增殖和凋亡的相关基因。结论 运用基因芯片技术筛选出的差异基因与PVDI所致大隐静脉曲张发病相关。     

腘静脉肌袢成形术治疗下肢深静脉瓣膜功能不全

目的研究下肢腘静脉肌袢成形术治疗下肢深静脉瓣膜功能不全的临床疗效. 方法 1997年1月～2001年6月,选择经过彩色多谱勒、静脉造影确诊为原发性下肢深静脉瓣膜功能不全的27例(27条)下肢,病程2～32年,平均17.4年.均有浅静脉曲张;其中小腿沉重感25例;肿胀22例;足靴区色素沉着25例;单侧慢性溃疡19例;2例曾行单纯大隐静脉结扎剥脱术.静脉压静息直立时平均为(11.00±0.73) kPa, 活动后平均为(9.14±0.68) kPa.均行腘静脉肌袢成形术,同时行大隐静脉高位结扎抽剥术,有12例行腔镜下交通支结扎. 结果术后27条患肢静脉压活动后平均下降为(5.94±0.82) kPa.与术前活动后平均值比较,差异有统计学意义(P<0.01).近期疗效良好,小腿沉重感及肿胀完全或基本消失,溃疡创面在2周内愈合.术后22条患肢获2～6年随访,21条患肢疗效优良,无复发;1例随访3年时因肌袢粘连导致小腿深静脉血栓形成. 结论严格掌握适应证,规范手术操作,腘静脉肌袢成形术治疗下肢深静脉瓣膜功能不全,可获得满意效果.     

静脉高压对大隐静脉和脾静脉管壁细胞形态学的影响

目的探讨静脉高压对大隐静脉和脾静脉管壁细胞形态的影响。方法分别收集曲张大隐静脉(20例)、门静脉高压性脾静脉管壁(14例)、正常大隐静脉(8例)和正常脾静脉(6例)标本,制作切片在电镜下观察其形态。结果电镜下曲张性大隐静脉和高压性脾静脉管壁均出现内皮细胞线粒体变性,平滑肌细胞胞浆内粗面内质网、核糖体、线粒体等细胞器增多,肌丝减少等改变。结论静脉高压影响曲张性大隐静脉和高压性脾静脉管壁细胞形态学的重塑,二者的改变是相同的。     

含血栓的曲张大隐静脉管壁肥大细胞的分布与定量研究

目的：探讨含血栓的曲张大隐静脉血管壁肥大细胞的浸润情况。方法：收集单纯曲张大隐静脉管壁标本11例为曲张组,含血栓的曲张大隐静脉管壁标本11例为血栓组,另设8例正常大隐静脉管壁标本为对照组,行甲苯胺蓝特殊染色,观察血管壁中肥大细胞浸润的分布与计数情况。结果：血栓组肥大细胞外膜明显增多,呈＂聚集征＂,中膜、内膜呈散在增多;曲张组肥大细胞外膜、中膜、内膜少量散在分布。血栓组管壁肥大细胞总计数与曲张组、对照组之间比较差异有统计学意义（P〈0.01）,曲张组与对照组之间比较差异无统计学意义（P〉0.05）。结论：静脉血栓形成和慢性炎症可能是导致血管壁肥大细胞大量浸润的重要因素。     

Update of endovenous treatment modalities for insufficient saphenous veins—A review of literature

Lower-limb venous insufficiency resulting from saphenous vein incompetence is a common disorder, increasing with age. For decades, surgical stripping of the great saphenous vein has been the gold standard in varicose vein treatment. The desire to optimize outcomes of treatment and reduce surgical trauma has led to the development of endovenous techniques. Today, several endovenous techniques are available to ablate the saphenous vein segments with abnormal vein valve function. In this review, we discuss the techniques, mechanisms of action, outcomes, and complications of all endovenous treatment modalities for the treatment of symptomatic lower-limb varicose veins.     

大隐静脉曲张管壁肥大细胞浸润分布与定量研究

目的探讨在不同病期、不同部位血管壁中肥大细胞浸润与大隐静脉曲张(VGSV)的关系。方法收集19例曲张大隐静脉的主干上、中、下3段管壁标本,其中曲张静脉伴有皮肤营养改变9例,单纯曲张静脉10例,另设9例正常静脉作对照,行甲苯胺蓝特殊染色。观察管壁中的肥大细胞浸润分布与计数情况。结果研究组肥大细胞的分布,内膜或中膜均可见少数肥大细胞,外膜则较多聚集。上、中、下3段血管壁肥大细胞计数结果:皮肤改变组(0.26±0.08)/HP、(0.24±0.18)/HP、(0.30±0.05)/HP;曲张组(0.26±0.12)/HP、(0.15±0.03)/HP、(0.12±0.05)/HP;对照组(0.23±0.14)/HP、(0.36±0.30)/HP、(0.27±0.16)/HP。各组间、段间的肥大细胞计数差异均无统计学意义。结论在不同病期、不同部位血管壁中肥大细胞的浸润㈦大隐静脉曲张的程度不存在量的变化关系。     

戴戒术与单纯浅静脉剥脱术治疗原发性下肢深静脉瓣膜功能不全的比较

目的:比较股浅静脉戴戒术与单纯性浅静脉剥脱术治疗下肢深静脉瓣膜功能不全的疗效。方法:将2010年03月—2011年1月收治的48例下肢深静脉瓣膜功能不全伴静脉曲张患者随机分为观察组(股浅静脉戴戒术联合大隐静脉高位结扎剥脱术)和对照组(传统大隐静脉高位结扎剥脱术),通过彩色多普勒检测两组术前及术后股总静脉血管内径、平均血流速度及Valsalva试验下反流持续时间的变化。结果:与术前比较,术后两组患者的静脉瓣膜功能及血流动力学均明显改善,表现为股总静脉管径缩小、平均血流速度增快、静脉血流反流时间缩短(均P<0.05),但观察组上述3个变量手术前后变化程度(差值)均明显大于照组(均P<0.05)。结论:两种手术方式均是治疗下肢深静脉瓣膜功能不全的有效手段,但戴戒术的治疗效果优于单纯性浅静脉剥脱术。     

复发性下肢静脉曲张再手术原因分析

目的:探讨分析复发性下肢静脉曲张再手术的原因。方法:总结63例复发性下肢静脉曲张患者的临床资料,应用彩色多普勒和顺行静脉造影了解下肢深静脉和交通支瓣膜功能。结果:63例中52例为下肢深静脉瓣膜功能不全,11例为交通支瓣膜功能不全伴隐静脉主干及其部分属支残留。结论:有效修复深静脉瓣膜功能,结扎和离断相应属支及交通支,阻断静脉的反流,纠正3个静脉系统及溃疡周围的血液滞留,是防治下肢静脉曲张术后复发的主要措施。     

Immunohistochemical and ultrastructural evidence that dendritic cells infiltrate stenotic aortocoronary saphenous vein bypass grafts

We earlier speculated that antigen-presenting dendritic cells may be involved in the immune reactions leading to saphenous vein bypass graft failure. The purpose of this study was to confirm whether dendritic cells are present in stenotic human saphenous vein bypass grafts. Segments of stenotic saphenous vein grafts were explanted from 14 patients at re-do bypass operation and ten normal saphenous veins were harvested during femoro-popliteal grafting. Sections of specimens were analysed using cell type specific antibodies to identify dendritic cells (CD1a, S-100), T-lymphocytes (CD3), macrophages (CD68), smooth muscle cells (alpha-SMA) and endothelial cells (FVIII). Dual immunostaining, confocal immunofluorescent laser scanning microscopy and electron microscopy were used. Stenotic grafts showed structural alterations of intimal hyperplasia and varying degrees of atherosclerotic degeneration. No cells expressing CD1a and S-100 were observed in the intima and media of normal saphenous veins. Cells expressing these antigens were present around areas of medial neovascularization and within intimal atherosclerotic lesions in saphenous vein bypass grafts. Electron microscopy demonstrated the presence of cells containing a well-developed tubulovesicular system which is unique to cells from the dendritic cell family. Double immunohistochemistry and confocal immunofluorescent microscopy revealed the co-localization of T-lymphocytes with dendritic cells. Dendritic cells are present in stenotic saphenous vein bypass grafts. Dendritic cells may be responsible for antigen presentation and modulation of immune reactions in accelerated graft atherosclerosis through their interaction with T-lymphocytes.     

下肢静脉曲张术后复发的原因分析(附46例报告)

目的分析下肢静脉曲张术后复发的原因。方法总结中山大学附属第二医院2002年6月至2004年8月收治的46例（52条患肢）下肢静脉曲张术后复发病人的临床资料，其中合并下肢静脉性溃疡20例（24条患肢）。所有病人均行深、浅静脉及交通静脉彩色多普勒超声检查和患肢顺行性深静脉造影。结果复发的原因有大隐静脉主干及其部分属支残存（11条患肢），合并交通静脉功能不全（35条患肢），合并深静脉瓣膜功能不全（20条患肢）。出现小隐静脉曲张（10条患肢），深静脉血栓形成后遗症（5条患肢），同时存在3个静脉系统病变（15条患肢）。结论下肢静脉曲张术后复发的原因较为复杂，通常累及3个静脉系统。交通静脉功能不全是导致下肢静脉曲张术后复发的主要原因。术前全面的影像学检查和选择正确的手术方式是避免术后静脉曲张复发的关键。     

下肢静脉高压对局部可溶性黏附分子水平表达的影响

目的研究下肢静脉高压对局部血清黏附分子含量的影响。方法经过双功彩超和静脉造影，选取单下肢慢性静脉功能不全（chronic venous insufficiency，CVI）的患者20例，在下肢静止下垂30min后分别抽取双下肢静脉血，同时取上肢静脉血作为对照组，而后取平卧位15min后再取双下肢静脉血。分为曲张静脉直立组（Vh组）、正常静脉直立组（Nh组）、曲张静脉平卧组（Vr组）、正常静脉平卧组（Nr组）和上肢对照组5组。用酶联免疫吸附法（ELISA）检测血清中的sICAM-1、sVCAM-1的含量。结果（1）Vh组和Nh组这两组静脉高压组的sICAM-1、sVCAM-1明显高于对照组（P〈0．01），而Vh组和Nh组之间差异无统计学意义（P〉0．05）。（2）Nr组和Nh组相比，sICAM-1、sVCAM-1均下降（P〈0．05）。（3）Vr组和Vh组相比，sICAM-1、sVCAM-1无明显变化（P〉0．05）。结论下肢静脉系统在血流淤滞和静脉高压的情况下，无论是否本身静脉有异常，局部的内皮细胞和白细胞均有激活并表达黏附分子。而在短时间静脉高压解除后，曲张静脉中局部的黏附分子高水平表达提示炎症仍持续，而正常静脉则有所下降。曲张静脉中炎症反应的持续性表达可能是CVI发展的重要环节。     

筋膜下内镜交通支离断术治疗下肢静脉性溃疡的体会

目的:探讨深筋膜下内镜交通支静脉离断术(subfascial endoscopic perforator surgery,SEPS)治疗下肢静脉性溃疡的可行性及应用价值。方法:回顾分析2008年12月至2011年4月为7例(8条肢体)下肢静脉性溃疡伴下肢静脉交通支瓣膜功能不全患者行大隐静脉剥脱术及SEPS的临床资料。结果:7例(8条肢体)手术均获成功,曲张静脉团块消失,下肢静脉血淤滞得到解决,色素沉着减轻,溃疡愈合。术后随访1～3年,无一例复发。结论:SEPS适于治疗下肢静脉交通支瓣膜功能不全的下肢静脉性溃疡患者,手术创伤小、安全、简单、易行,疗效确切,应用价值较高。     

An animal model of venous hypertension: the role of inflammation in venous valve failure

BACKGROUND: Clinical observation suggests that chronic venous insufficiency is related to failure of venous valves. Duplex ultrasound studies of lower extremity superficial veins regularly show valve failure and venous reflux. Gross morphologic observation of venous valves in surgical specimens shows tearing, splitting, scarring, and disappearance of valves. HYPOTHESIS: Venous valve damage is acquired, linked with venous hypertension, and affected by inflammation. OBJECTIVE: The objective of this study was to investigate the inflammatory process in valve remodeling associated with acute and chronic venous hypertension. METHODS: A femoral arteriovenous fistula was created in study animals (Wistar rats, n = 60), and animals without an arteriovenous fistula were studied as controls (n = 5). At 1, 7, 21, and 42 days animals with the femoral arteriovenous fistula were anesthetized, and systemic pressure, the pressure in the femoral vein distal to fistula, and the pressure of the femoral vein in the contralateral hind limb were measured. Timed collection of blood backflow after division of the femoral vein distal to the fistula and in the alive, anesthetized animal was collected, measured, and calculated per unit time to be used as an indicator of valve insufficiency. The femoral vein distal to the fistula was harvested; valvular structures were examined and measured. Specimens were processed, and longitudinal sections were made and challenged with immunostaining antibodies against matrix metalloprotease (MMP)-2 and MMP-9. Sections were examined, and expression of molecular markers was determined by light absorption measurements after image digitization. RESULTS: One week after the procedure, all animals exhibited some degree of hind limb edema ipsilateral to the arteriovenous fistula. Pressure in the femoral vein distal to the fistula was markedly increased on average to 96 +/- 9 mm Hg. Reflux was increased in a time-dependent manner, with the 21-day and 42-day groups showing the highest values. Valves just distal to the fistula showed an increased diameter of the valvular annulus and a shortening of the annular height. Venous wall findings included fibrosis and fusion of the media and adventitia and scarring and disappearance of valves principally in the 21- and 42-day specimens. Immunolabeling for MMP-2 showed an increased level in the 21- and 42-day groups. MMP-9 showed an increased level at 1 day, followed by a more marked level in the 21- and 42-day groups. CONCLUSIONS: In this animal model of venous hypertension the findings of limb edema, increasing valvular reflux, and morphologic changes of increased annulus diameter and valve height are seen. Histologic changes included massive fibrosis of media and fusion with adventitia. Inflammatory markers MMP-2 and MMP-9 are strongly represented, and valve disappearance occurs after these markers are present. The gross morphologic changes seen are quite similar to those observed in human surgical specimens removed in treatment of venous insufficiency. CLINICAL RELEVANCE: When observed angioscopically at the time of vein stripping, saphenous vein valves show severe deformities including shortening, scarring, and tearing. The current model of induced venous hypertension demonstrates early venous valve changes that replicate those observed in humans. This observation provides a link from venous hypertension to an induced inflammatory reaction that stimulates the valve damage. Thus the model could be useful for defining the fundamental mechanisms that cause venous valve failure and varicose veins and in pharmacologic testing to prevent or treat venous insufficiency.     

Venous morphology predicts class of chronic venous insufficiency.

OBJECTIVES: this study aimed to determine specific morphological differences in long saphenous veins from patients with various grades of chronic venous insufficiency. DESIGN: comparable veins from a control group were compared with patients with either primary varicose veins or those with associated skin changes including venous ulcers. MATERIALS: below-knee segments of saphenous vein were examined from a total of 64 patients. METHODS: veins were examined for elastic-tissue disruption and endothelial-cell changes and comparisons made between clinical groups. RESULTS: elastic-tissue disruption, as measured by fragmentation of the elastic lamina and the percentage of the intimal-medial boundary containing elastin, increased with increasing severity of venous disease. Moreover, endothelial cells became more densely packed, as measured by endothelial cell and endothelial-cell nuclei density, with increasing severity of disease. Other measures such as the density of multinucleated "giant" endothelial cells and the number of nuclei per "giant" cell did not correlate with venous disease, however. CONCLUSIONS: this study demonstrates that several morphological characteristics of superficial saphenous veins correlate with severity of venous disease. In particular, the alterations to the structure of elastic tissue within these veins appears indicative of the progressive nature of chronic venous insufficiency.     

Femoral venous reflux abolished by greater saphenous vein stripping

Preoperative venous duplex scanning has revealed unexpected deep venous incompetence in patients with apparently only varicose veins. Acting on the hypothesis that the deep vein reflux was secondary to deep vein dilation caused by reflux volume, the following was done. Between July 1990 and April 1993, 29 limbs in 21 patients (16 females) were examined by color-flow duplex imaging to determine valve closure by the method of van Bemmelen. Instrumentation included high-resolution ATL-9 venous interrogation using a pneumatic cuff deflation stimulus of reflux in the standing, nonweight-bearing limb. All limbs showed greater saphenous vein reflux. Twenty-nine showed superficial femoral vein reflux and of these three showed popliteal vein reflux. Duplex testing was performed by a certified vascular technologist whose interpretation was blinded as to the results of clinical examination and grading of the severity of venous insufficiency. Surgery was performed on an outpatient basis under general anesthesia using groin-to-knee removal of the greater saphenous vein by the vein inversion technique of Van Der Strict. Stab avulsion of varicose tributary veins was accomplished during the same period of anesthesia. In 27 of 29 limbs with preoperative femoral reflux, that reflux was abolished by greater saphenous stripping. In patients with popliteal reflux both femoral and popliteal reflux was abolished. Improvement of deep venous hemodynamics by ablation of superficial reflux supports the reflux circuit theory of venous overload. Furthermore, preoperative evaluation of venous hemodynamics by duplex scanning appears to provide useful pre- and postoperative information regarding venous insufficiency in individual patients.Presented at the Twelfth Annual Meeting of the Southern California Vascular Surgical Society, Coronado, Calif, September 17–19, 1993.     

含血栓静脉管壁病理形态学特征的比较性研究

目的 探讨含血栓的静脉管壁病理形态学特征及管腔内径变化.方法 收集含血栓的曲张大隐静脉主干标本18例(血栓组),单纯曲张大隐静脉主干标本18例(曲张组),另设正常大隐静脉主干标本12例(对照组).采用苏木精-伊红和Masson染色法,观察管壁组织结构,测量血管壁内径与内膜、中膜、外膜厚度.结果 血栓组管壁全层及血栓内见大量炎细胞浸润,血栓附着处内皮细胞缺失,大量增生的胶原纤维自破裂内膜处延伸至血栓内致部分血栓机化.曲张组管壁全层偶见炎细胞,内膜增厚,中膜平滑肌细胞不规则增生,排列紊乱,胶原纤维增生.对照组管壁内膜薄,中膜平滑肌束排列规则.血栓组管腔内径、内膜厚度、中膜厚度和外膜厚度与曲张组、对照组比较差异均有统计学意义(P＜0.05),曲张组管腔内径和内膜厚度与对照组比较差异有统计学意义(P＜0.05).结论 含血栓的曲张静脉管腔增大、管壁增厚是病理形态学改变的主要特征.