Free amino acids in normal and varicose human saphenous veins.

An increase in the activity of lysosomal enzymes in varicose veins has been suggested in the literature, as well as an alteration of collagen fibrils situated near muscle cells. In view of these results, we have compared the free amino acid contents of varicose veins with those of healthy veins removed few hours after death. The amino acid analyses of extracts have shown that the varicose vein appears to contain about 2.7 times more free amino acids than normal one, except in the case of phosphoethanolamine. Controls have shown that this discrepancy can be ascribed to a rapid liakage of the amino acids after death. The absence of hydroxyproline suggests that collagen does not seem liable to degradation in the varicose vein. On the other hand, the constant phosphoethanolamine content found can be attributed to a much higher concentration of this compound in the varicose vein or alternatively, to a postmortem process of degradation, compensating for the losses due to diffusion.     

IGFBP6 regulates vascular smooth muscle cell proliferation and morphology via cyclin E–CDK2

Despite the high prevalence of varicose veins, the underlying pathogenesis of this disease remains unclear. The present study aims to explore the role of insulin-like growth factor binding protein 6 (IGFBP6) in vascular smooth muscle cells (VSMCs). Using a protein array approach, we identified several differentially expressed proteins between varicose great saphenous veins and normal great saphenous veins. Bioinformatic analysis showed that IGFBP6 was closely related to cell proliferation. Further validation confirmed that IGFBP6 was one of the most highly expressed proteins in varicose vein tissue. Knocking down IGFBP6 in VSMCs significantly attenuated cell proliferation and induced the S phase arrest during the cell cycle. Further experiments demonstrated that IGFBP6 knockdown increased cyclin E ubiquitination, which reduced expression of cyclin E and phosphorylation of CDK2. Furthermore, IGFBP6 knockdown arrested centrosome replication, which subsequently influenced VSMC morphology. Ultimately, IGFBP6 was validated to be involved in VSMC proliferation in varicose vein tissues. The present study reveals that IGFBP6 is closely correlated with VSMC biological function and provides unprecedented insights into the underlying pathogenesis of varicose veins.     

改良曲张静脉点式剥除术治疗中老年下肢静脉曲张的临床疗效

目的:探究改良曲张静脉点式剥除术在治疗中老年下肢静脉曲张的临床疗效。方法:收集我院已确诊为下肢静脉曲张的中老年患者37例,分成实验组与对照组。对照组18例行传统曲张静脉点式剥除术,实验组19例行改良曲张静脉点式剥除术。对比两组患者手术后的下肢静脉曲张的治疗效果。结果:实验组有效率(94.7%)显著高于对照组(72.2%),差异具有统计学意义(P0.05);与对照组相比,实验组患者手术时间较短、术中出血量较少、下床活动时间较早,术后并发症总治愈率较高,复发率、术后并发症发生率较低,其差异均有统计学意义(P0.05)。结论:采用改良曲张静脉点式剥除术治疗中老年下肢静脉曲张的患者能够更彻底的剥除曲张额静脉,有效的改善患肢症状,明显降低复发率。     

The determination of the collagen and elastin amount in the human varicose vein by the computer morphometric method

The results of the works dealing with alterations of the connective tissue in varicose vein wall are not ambiguous, so the exact cause of the vein dilatation has still not been established. We were determining the collagen and elastin amounts in human varicose vein wall in comparison with non-dilated long saphenous vein by the light microscopy and computer morphometric method. We have found the lesser amount of collagen in varicose veins than in non-dilated veins, the amounts of the elastin in both the varicose and non-varicose veins were without the statistical significance.     

Influence of thrombophlebitis on TGF-β1 and its signaling pathway in the vein wall

Extensive extracellular matrix remodeling of the vein wall is involved in varicose veins pathogenesis. This process is controlled by numerous factors, including peptide growth factors. The aim of the study was to evaluate influence of thrombophlebitis on TGF-β1 and its signaling pathway in the vein wall. TGF-β1 mRNAlevels, growth factor content and its expression were evaluated by RT-PCR, ELISA, and western blot methods, respectively, in the walls of normal veins, varicose veins and varicose veins complicated by thrombophlebitis. Western blot analysis was used to assess TGF-β receptor type II (TGF-β RII) and p-Smad2/3 protein expression in the investigated material. Unchanged mRNA levels of TGF-β1, decreased TGF-β1 content, as well as decreased expression of latent and active forms of TGF-β1 were found in varicose veins. Increased expression of TGF-β RII and p-Smad2/3 were found in varicose veins. Thrombophlebitis led to increased protein expression of the TGF-β1 active form and p-Smad2/3 in the vein wall compared to varicose veins. TGF-β1 may play a role in the disease pathogenesis because of increased expression and activation of its receptor in the wall of varicose veins. Thrombophlebitis accelerates activation of TGF-β1 and activity of its receptor in the varicose vein wall.     

White cell accumulation in dependent legs of patients with venous hypertension: a possible mechanism for trophic changes in the skin

The mechanism by which chronic venous insufficiency and venous hypertension are associated with ulceration of the legs is not yet understood. To investigate this mechanism further accumulation of white cells in the dependent legs of normal volunteers, patients awaiting surgery for simple varicose veins, and patients with chronic venous insufficiency was studied. About 24% fewer white cells than in normal subjects left the dependent foot of patients with venous hypertension, and this trapping of white cells, was reversed when the foot was raised; similar changes were not observed in normal subjects or patients with varicose veins.The trophic skin changes typically seen in patients with venous hypertension may be aggravated by damage caused by the repeated accumulation of white cells in the microcirculation.     

2 types of dynamics of filling of veins in the extremities during obstruction of venous outflow

The method of venous occlusion plethysmography improved by introduction of differential and logarithmic-computing signal amplifiers was used to study the time course of blood inflow to the limbs in 35 healthy individuals and 115 patients with diseases of lower limb veins. Two types of the time course of the vein blood content were defined. With type I the volume velocity (VV) of blood supply to the segment under study decreases, whereas with type II the VV first decreases and then remains unchanged for a long time. Type I occurs in 76% of healthy individuals, type II in 74% of patients. With type II the increment of the vein capacity is significantly reduced in both healthy individuals and in patients with varicose veins. Moreover, in healthy individuals, the VV, the time of constant venous volume attainment, and the rate of blood outflow from veins (after occlusion removal) significantly differ as well in types I and II. Both types of the blood content remain unchanged while both applying the functional tests and during examinations at varying times. It is assumed that type II blood content is determined by greater rigidity of the venous vessels.     

Augmentation of miR-202 in varicose veins modulates phenotypic transition of vascular smooth muscle cells by targeting proliferator–activated receptor-γ coactivator-1α

In varicose veins, vascular smooth muscle cells (VSMCs) often show abnormal proliferative and migratory rates and phenotypic transition. This study aimed to investigate whether microRNA (miR)-202 and its potential target, peroxisome proliferator–activated receptor-γ coactivator-1α (PGC-1α), were involved in VSMC phenotypic transition. miR-202 expression was analyzed in varicose veins and in VSMCs conditioned with platelet-derived growth factor. The effect of miR-202 on cell proliferation and migration was assessed. Furthermore, contractile marker SM-22α, synthetic markers vimentin and collagen I, and PGC-1α were analyzed by Western blot analysis. The modulation of PGC-1α expression by miR-202 was also evaluated. In varicose veins and proliferative VSMCs, miR-202 expression was upregulated, with decreased SM-22α expression and increased vimentin and collagen I expression. Transfection with a miR-202 mimic induced VSMC proliferation and migration, whereas a miR-202 inhibitor reduced cell proliferation and migration. miR-202 mimic constrained luciferase activity in HEK293 cells that were cotransfected with the PGC-1α 3′-untranslated region (3′-UTR) but not those with mutated 3′-UTR. miR-202 suppressed PGC-1α protein expression, with no influence on its messenger RNA expression. PGC-1α mediated VSMC phenotypic transition and was correlated with reactive oxygen species production. In conclusion, miR-202 affects VSMC phenotypic transition by targeting PGC-1α expression, providing a novel target for varicose vein therapy.     

Subpopulations of human T lymphocytes. X. Alterations in T, B, third population cells, and T cells with receptors for immunoglobulin M (Tmu) or G (Tgamma) in aging humans.

Peripheral blood from 120 healthy subjects, of whom 59 were young (35.5 +/- 9.6 years) and 61 aging (69.2 +/- 4.2 years), was examined for the proportions and numbers of lymphocyte populations, with a battery of surface markers. Absolute lymphocyte count and T,B and third population cells were comparable in both groups. Tgamma cell proportions were significantly (p less than 0.001) increased in aging subjects when compared with the young subjects. However, this difference was more significant (p less than 0.001) when aged females were compared with the young females as compared to aged males vs young males (p less than 0.05). When data were anlayzed for absolute numbers of Tmu and Tgamma cells, a similar significant decrease in Tmu, and increase in Tgamma cells were observed. Interestingly, when these data were anlayzed according to gender, significant differences in Tmu and Tgamma cell number were observed between young and old females but not between young and old men. Implications of these results are discussed.     

Generation of reactive oxygen species by a sufficient, insufficient and varicose vein wall

Despite numerous theories, the etiology and pathogenesis of primary varicose veins remain unclear. The etiology of chronic venous diseases (CVDs) known as chronic venous insufficiency (CVI) is related to leukocyte trapping. Leukocyte trapping involves trapping of white cells in vessel walls followed by their activation and translocation outside the vessel. Release of reactive oxygen species (ROS) from trapped white cells has been documented. Superoxide dismutase (SOD) directly inhibits the generation of free radicals and compounds that are produced during oxidation by ROS, such as malonyldialdehyde (MDA). The aim of this study was to determine the involvement of free radicals in the etiology of venous changes. The following material was used for the study: fragments of sufficient or insufficient venous system and varices from 31 patients diagnosed with chronic venous disease in the 2nd or 3rd degree, according to clinical state, etiology, anatomy and pathophysiology (CEAP), which were qualified for surgical procedure. The levels of oxidative stress markers strongly correlated with lesions observed by USG in insufficient and varicose veins. In both a higher concentration of MDA was observed, which is a sign of lipid peroxidation. Antioxidative mechanisms, SOD activity and total antioxidative power expressed as FRAP were inversely proportional to MDA concentration. In insufficient and varicose veins both FRAP and SOD activities were significantly lower than in normal veins. The severity of clinical changes was inversely dependent on the efficiency of scavenging of ROS, which additionally proves the participation of free radicals in pathogenesis of CVDs.     

A high accumulation of minerals in human internal jugular vein

To elucidate age-related changes of mineral contents in human veins, the relative contents (RCs) of elements in the human internal jugular veins, superior and inferior venae cavae, and femoral veins from 27 subjects ranging from 40 to 98 yr old were analyzed by inductively coupled plasma atomic emission spectrometry. The average RCs of calcium in the 27 specimens were the highest in the internal jugular vein, followed in descending order by the superior vena cava, femoral vein, and inferior vena cava. The RCs of calcium and phosphorus in the internal jugular veins started to increase after the age of 50 yr, became the highest in subjects in their 60s and thereafter decreased gradually. It is noted that such accumulations of minerals, similar to the internal jugular vein, were also found in the veins, such as the superior and inferior venae cavae, and femoral vein. Accumulation of minerals in the veins is different from that of arteries, which increase progressively by aging, but do not decrease.     

Compression-sclerotherapy for varicose veins: a Canadian study.

In recent years modifications and refinements in treating varicose veins by injecting a sclerosant have given this method an important place in the practitioner''s armamentarium. Specific indications and precise technique are essential. Compression-sclerotherapy has been used since 1975 in the vein clinic of Belleville General Hospital, Belleville, Ont. Some patients have also required limited ligation of veins that were deeper or more proximal, or both. A follow-up study of patients 2 or more years after treatment revealed no major complications and a high rate of patient acceptance and satisfaction. The costs of treatment were about one tenth those of conventional inpatient surgery.     

Influenza-specific T cells from older people are enriched in the late effector subset and their presence inversely correlates with vaccine response

T cells specific for persistent pathogens accumulate with age and express markers of immune senescence. In contrast, much less is known about the state of T cell memory for acutely infecting pathogens. Here we examined T cell responses to influenza in human peripheral blood mononuclear cells from older (>64) and younger (<40) donors using whole virus restimulation with influenza A (A/PR8/34) ex vivo. Although most donors had pre-existing influenza reactive T cells as measured by IFNγ production, older donors had smaller populations of influenza-responsive T cells than young controls and had lost a significant proportion of their CD45RA-negative functional memory population. Despite this apparent dysfunction in a proportion of the older T cells, both old and young donors' T cells from 2008 could respond to A/California/07/2009 ex vivo. For HLA-A2+ donors, MHC tetramer staining showed that a higher proportion of influenza-specific memory CD8 T cells from the 65+ group co-express the markers killer cell lectin-like receptor G1 (KLRG1) and CD57 compared to their younger counterparts. These markers have previously been associated with a late differentiation state or immune senescence. Thus, memory CD8 T cells to an acutely infecting pathogen show signs of advanced differentiation and functional deterioration with age. There was a significant negative correlation between the frequency of KLRG1(+)CD57(+) influenza M1-specific CD8 T cells pre-vaccination and the ability to make antibodies in response to vaccination with seasonal trivalent inactivated vaccine, whereas no such trend was observed when the total CD8(+)KLRG1(+)CD57(+) population was analyzed. These results suggest that the state of the influenza-specific memory CD8 T cells may be a predictive indicator of a vaccine responsive healthy immune system in old age.     

VARICOSE VEINS—A Practical Approach to Treatment

Adequate treatment of varicose veins requires thorough mapping of perforating veins, communicating veins and “blow out” areas. Combined ligations, stripping and injection of sclerotic substances after operation is the most effective regimen of therapy.The technique of stripping is facilitated by isolating the saphenous vein at the ankle, inserting the stripper from below upward, then making a transverse groin incision over the palpable stripper. The tip of the stripper should be twice the diameter of the vein to be removed. Stripping should be done with the patient in the Trendelenburg position.All patients must be examined at regular intervals after operation and injection of sclerosing material carried out as necessary.     

Role of mechanosignaling on pathology of varicose vein

Varicose veins are the most common vascular disease in humans. Veins have valves that help the blood return gradually to the heart without leaking blood. When these valves become weak, blood and fluid collect and pool by pressing against the walls of the veins, causing varicose veins. In the cardiovascular system, mechanical forces are important determinants of vascular homeostasis and pathological processes. Blood vessels are constantly exposed to a variety of hemodynamic forces, including shear stress and environmental strains caused by the blood flow. In varicose veins within the leg, venous blood pressure rises in the vein of the lower extremities due to prolonged standing, creating a peripheral tension in the vessel wall thereby causing mechanical stimulation of endothelial cells and vascular smooth muscle. Studies have shown that long-term increased exposure to vascular wall tension is associated with the overexpression of HIF-1α and HIF-2α and increased levels of MMP-2 and MMP-9, thereby reducing venous contraction and progressive venous dilatation, which is involved in the development of varicose veins. Following the expression of metalloproteinase, the expression of type 1 collagen increases, and the amount of type 3 collagen decreases. Therefore, collagen imbalance will cause the varicose veins to not stretch. Loss of structural proteins (type 3 collagen and elastin) in the vessel wall causes the loss of the biophysical properties of the varicose vein wall. This review article tries to elaborate on the effect of mechanical forces and sensors of these forces on the vascular wall in creating the mechanism of mechanosignaling, as well as the role of the onset of molecular signaling cascades in the pathology of varicose veins.     

Assessment of S100 protein expression in the epididymis of juvenile and adult European bison

In our study, we decided to compare S100 protein expression in the material obtained from the epididymes of 5- and 12-month-old calves, and adult European bison, and to detect any differences in S100 expression according to the animal age and size of the organ examined. We used the epididymes obtained from 6 adult European bison aged 6-12 years, from 6 at the age of 12 months and 6 calves aged 5 months. Immunocytochemical reactions were performed using the avidin-biotinylated-peroxidase (ABC) technique according to HSU. Specific polyclonal rabbit antiserum against bovine S100 protein (Bio Genex Laboratories) at a dilution at 1:400 was applied. We found the expression of S100 protein in endothelial cells of arteries, veins and lymphatic vessels in all the study animals. At the same time, we found no differences in the expression of S100 protein in vascular endothelial cells. Our observations seem to indicate that S100 expression in endothelial cells of European bison epididymis is not correlated with age or maturity of the organ tested. We found S100 protein in smooth muscle cells of arteries and veins in all European bison specimens examined. Interestingly in the current study, in young 5-month-old sexually immature European bison specimens we observed weaker expression of S100 protein in smooth muscle cells of small vessels as compared to the same cell type both in large vessels in these animals and in small vessels in adult specimens.     

The elastic fibre system in the veins of the human lower oesophagus

The elastic, elaunin and oxytalan fibres in the tunica mucosa and tela submucosa of veins of the human lower oesophagus were studied in 30 necropsy and biopsy specimens using appropriate histological and ultrastructural methodologies. Elaunin fibres predominate in the veins endowed with muscle cells. Scattered oxytalan and elastic fibres were also observed, the latter being more numerous at the vein periphery. No noteworthy fibre arrangement was encountered in veins lacking muscle cells. Those fibres disposed around the veins were considered to belong to the connective tissue of the tunica mucosa and tela submucosa in which they are embedded. The veins of the lower oesophagus seem to be of low elasticity, which may be related to a blockage mechanism described for the gastro-oesophageal blood stream in cases of portal hypertension.     

Regional differences in the developing foreleg of Drosophila melanogaster

We transplanted imaginal disks of Drosophila melanogaster from larvae of the second half of the third larval instar into prepupae. Disks from the youngest donors differentiated bristles of only the distal segments of the leg. These disks also produced unusually large areas of cuticle that had no bristles. Disks from older donors differentiated bristles of more proximal segments and the area of cuticle with no bristles was reduced. To account for the regional variation in these results, there must be regional differences among the prospective leg cells at some time during the period from the second half of the third larval instar to the end of adult bristle differentiation. We asked whether prospective distal cells were more advanced than prospective proximal cells during bristle differentiation. We estimated when bristle precursor cells undergo their final cell divisions by heavily irradiating prepupae and pupae. We assumed that cells that were insensitive to the radiation had completed their cell divisions. The distal segments were the first to have insensitive bristles. Most leg bristles became insensitive between 12 and 18 hr after pupariation. The tarsus had a larger proportion of its bristles insensitive than the femur at 15 hr after pupariation. We also investigated when bristle-forming cells begin elongating their bristle shafts. We used the length of bristle rudiments as an indicator of when elongation is initiated. At 35 hr after pupariation, bristle rudiments of distal segments were two to three times longer than bristle rudiments of proximal segments. We discuss how these intersegmental differences observed during bristle differentiation can account for the regional variation in response of discs transplanted into older hosts. However, we do not exclude the possibility that regional differences among cells of the leg tissue exist at stages earlier than the time of bristle differentiation.     

Varicose veins; a practical approach to treatment

Adequate treatment of varicose veins requires thorough mapping of perforating veins, communicating veins and "blow out" areas. Combined ligations, stripping and injection of sclerotic substances after operation is the most effective regimen of therapy. The technique of stripping is facilitated by isolating the saphenous vein at the ankle, inserting the stripper from below upward, then making a transverse groin incision over the palpable stripper. The tip of the stripper should be twice the diameter of the vein to be removed. Stripping should be done with the patient in the Trendelenburg position. All patients must be examined at regular intervals after operation and injection of sclerosing material carried out as necessary.