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Introduction: The disease arteriosclerosis obliterans (ASO) affects the lower extremities. ASO's mechanism involves the proliferation and migration of vascular smooth muscle cells (VSMCs). The miR-4284 is involved in several biological processes of the cardiovascular system, including VSMC proliferation, migration, and death. However, it is unknown if the miR-4284 gene is involved in the control of ASO. Furthermore, the molecular processes behind the contribution of human arterial smooth muscle cells (HASMCs), one of the most significant components of the arterial wall, to arteriosclerosis obliterans (ASO) pathogenesis remain unknown. Previously, we explored the alterations of miRNAs in the blood of ASO patients, and now we wanted to test further whether these changes also take place in the HASMCs that are responsible for the pathogenesis of ASO., Methods: The expression levels of miR-29a in arterial walls were analyzed via a real-time polymerase chain reaction. An ASO cell model was established to investigate the expression of miR- 4284 on HASMCs. The Transwell system and CCK-8 detection were used to assess the migration and proliferation of HASMCs. The proportion of apoptotic cells as well as the concentrations of apoptotic signal protein production were assessed using flow cytometry. A Western blot technique was used to identify B cell lymphoma-2 (Bcl2), Bcl2-associated X protein (BAX), as well as Xlinked inhibitors of apoptosis protein (XIAP)., Results: The results showed that PCR confirmed that the qualified production or expression of miR-4284 was significantly reduced in HASMCs after they were cultured without FBS and in an atmosphere of 1% O 2 + 5% CO 2 + 94% N 2 and that glucose had no effect on its expression. MiR- 4284 has no effect on migration and proliferation, but downregulation of miR-4284 can decrease the apoptotic rate of HASMCs, as revealed by flow cytometry. Furthermore, western blot experiments showed that the expression of BAX was low, while the expression of the other two proteins, viz., Bcl2 and XIAP, was over-expressed., Conclusion: We found that miR-4284 downregulation enhanced Bcl2, as well as XIAP, and decreased Bax. This shows that downregulated miR-4284 regulates apoptosis-related protein expression in HASMCs. The mechanism is not clear, and we need further study to confirm it., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Context: Qing-Mai-Yin (QMY) is a clinically used herbal formula for treating arteriosclerosis obliterans (ASO)., Objective: To evaluate the chemical constituents and effects of QMY on ASO rabbit model., Materials and Methods: Forty-eight New Zealand rabbits were divided into six groups ( n  = 8): normal (normal rabbits treated with 0.5% CMC-Na), vehicle (ASO rabbits treated with 0.5% CMC-Na), positive (simvastatin, 1.53 mg/kg), and QMY treatment (300, 600, and 1200 mg/kg). ASO rabbit model was prepared by high fatty feeding, roundly shortening artery, and bovine serum albumin immune injury. QMY (300, 600 and 1200 mg/kg) was orally administered for 8 weeks. The effects and possible mechanisms of QMY on ASO rabbits were evaluated by pathological examination, biochemical assays, and immunohistochemical assays. The compositions of QMY were analysed using HPLC-Q-TOF-MS/MS analysis., Results: Compared to the vehicle rabbit, QMY treatment suppressed plaque formation and intima thickness in aorta, and decreased intima thickness, whereas increased lumen area of femoral artery. Additionally, QMY treatment decreased TC, TG and LDL, decreased CRP and ET, and increased NO and 6-K-PGF1α in serum. Furthermore, the potential mechanisms studied revealed that QMY treatment could suppress expression of TNF-α, IL-6, ICAM-1 and NF-κB in endothelial tissues, and increase IκB. In addition, HPLC analysis showed QMY had abundant anthraquinones, stilbenes, and flavonoids., Conclusion: QMY has ameliorative effects on ASO rabbit, and the potential mechanisms are correlated to reducing inflammation and down-regulating NF-κB. Our study provides a scientific basis for the future application and investigation of QMY.

Objective: To explore the effect of long non-coding ribonucleic acid (lncRNA) H19 on the apoptosis of vascular endothelial cells in arteriosclerosis obliterans (ASO) via the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway., Patients and Methods: Human umbilical vein endothelial cells (HUVECs) were cultured, and lncRNA H19 was inhibited by Si-H9 and overexpressed by H19-OE. Then, the apoptosis rate was detected by flow cytometry, the target of lncRNA H19 was detected by dual luciferase reporter gene assay, and changes in the protein level were determined via Western blotting (WB)., Results: LncRNA H19 exhibited high expression in serum of patients with ASO, and compared with that in congeneric normal mice, the expression of lncRNA H19 in ASO mice rose. Besides, the proliferation ability of cells transfected with H19-OE was markedly strengthened, and H19-OE treatment could down-regulate the expression level of the apoptin, active cysteinyl aspartate-specific proteinase-3 (Caspase-3). In addition, lncRNA H19 bound to micro ribonucleic acid (miR)-19a in a targeted way. After lncRNA H19 was overexpressed, the expression of the NF-κB pathway key factors, p38 and p65, were notably increased, and the nuclear translocation of p65 was significantly enhanced after transfection with miR-19a., Conclusions: LncRNA H19 promotes the proliferation of vascular endothelial cells in ASO and inhibits the apoptosis of them via the NF-κB pathway.

Aberrant vascular smooth muscle cell (VSMC) migration has been implicated in a variety of vascular disorders, while the signal pathways governing this process remain unclear. Here, we investigated whether miRNAs, which are strong post-transcriptional regulators of gene expression, could alter VSMC migration. We detected the expression of miR-4463 in the plasma of patients with atherosclerosis and in human aortic smooth muscle cells under hypoxia-ischemia condition, and investigated the migration effect and its downstream pathways. The results have shown that whether in clinical AS patients or hypoxic cells, the expression of miR-4463 was lower than that of normal group, then the number of migrating cells in the miR-4463 mimic intervention group was significantly decreased compared with the normal group and miR-4463 inhibitor instead. Furthermore, the expression of angiomotin (AMOT) in gastrocnemius muscle and femoral artery of patients was significantly higher than that of the control group. The protein level of AMOT in miR-4463 mimic intervention group was significantly decreased, and its level was reversed by inhibiting miR-4463. In summary, these results indicate that miR-4463 is a novel modulator of VSMC migration by targetting AMOT expression. Regulating miR-4463 or its specific downstream target genes in VSMCs may represent an attractive approach for the treatment of vascular diseases., (© 2018 The Author(s).)

Background/aims: Increasing evidence links microRNAs to the pathogenesis of peripheral vascular disease. We recently found microRNA-125b (miR-125b) to be one of the most significantly down‑regulated microRNAs in human arteries with arteriosclerosis obliterans (ASO) of the lower extremities. However, its function in the process of ASO remains unclear. This study aimed to investigate the expression, regulatory mechanisms, and functions of miR-125b in the process of ASO., Methods: Using the tissue explants adherent method, vascular smooth muscle cells (VSMCs) were prepared for this study. A rat carotid artery balloon injury model was constructed to simulate the development of vascular neointima, and a lentiviral transduction system was used to overexpress serum response factor (SRF) or miR-125b. Quantitative real‑time PCR (qRT‑PCR) was used to detect the expression levels of miR‑125b and SRF mRNA. Western blotting was performed to determine the expression levels of SRF and Ki67. In situ hybridization analysis was used to analyze the location and expression levels of miR-125b. CCK-8 and EdU assays were used to assess cell proliferation, and transwell and wound closure assays were performed to measure cell migration. Flow cytometry was used to evaluate cell apoptosis, and a dual-luciferase reporter assay was conducted to examine the effects of miR‑125b on SRF. Immunohistochemistry and immunofluorescence analyses were performed to analyze the location and expression levels of SRF and Ki67., Results: miR-125b expression was decreased in ASO arteries and platelet-derived growth factor (PDGF)-BB-stimulated VSMCs. miR-125b suppressed VSMC proliferation and migration but promoted VSMC apoptosis. SRF was determined to be a direct target of miR-125b. Exogenous miR-125b expression modulated SRF expression and inhibited vascular neointimal formation in balloon-injured rat carotid arteries., Conclusions: These findings demonstrate a specific role of the miR-125b/SRF pathway in regulating VSMC function and suggest that modulating miR-125b levels might be a novel approach for treating ASO., (© 2018 The Author(s). Published by S. Karger AG, Basel.)

Results of the hemostasis conduction in conditions of revascularization in 106 patients, оperated on for atherosclerotic affection of aorta and the main arteries of the lower extremities, were adduced. Syndrome of hypercoagulation of traumatic stage of surgical intervention in early postoperative period is developing due to thrombinemia on background of a fibrinolytic system depression. There was proved a necessity to impact on thrombin-fibrinous factor (factor ІІа) of hemocoagulant cascade by application of nonfractionized heparins immediately after conclusion of operative intervention with thromboprophylaxis prolongation, using low-molecular heparins (impact on Ха factor) in accordance to the branch standards.

Background: Aberrant vascular smooth muscle cell (VSMC) proliferation and migration contribute to the development of vascular pathologies, such as atherosclerosis and post-angioplasty restenosis. The aim of this study was to determine whether miR-22-3p plays a role in regulating human artery vascular smooth muscle cell (HASMC) function and neointima formation., Methods: Quantitative real-time PCR (qRT-PCR) and fluorescence in situ hybridization (FISH) were used to detect miR-22-3p expression in human arteries. Cell Counting Kit-8 (CCK-8) and EdU assays were performed to assess cell proliferation, and transwell and wound closure assays were performed to assess cell migration. Moreover, luciferase reporter assays were performed to identify the target genes of miR-22-3p. Finally, a rat carotid artery balloon-injury model was used to determine the role of miR-22-3p in neointima formation., Results: MiR-22-3p expression was downregulated in arteriosclerosis obliterans (ASO) arteries compared with normal arteries, as well as in platelet-derived growth factor-BB (PDGF-BB)-stimulated HASMCs compared with control cells. MiR-22-3p overexpression had anti-proliferative and anti-migratory effects and dual-luciferase assay showed that high mobility group box-1 (HMGB1) is a direct target of miR-22-3p in HASMCs. Furthermore, miR-22-3p expression was negatively correlated with HMGB1 expression in ASO tissue specimens. Finally, LV-miR-22-3p-mediated miR-22-3p upregulation significantly suppressed neointimal hyperplasia specifically by reducing HMGB1 expression in vivo., Conclusions: Our results indicate that miR-22-3p is a key molecule in regulating HASMC proliferation and migration by targeting HMGB1 and that miR-22-3p and HMGB1 may be therapeutic targets in the treatment of human ASO., (© 2017 The Author(s). Published by S. Karger AG, Basel.)

Background and Aims: Cholesterol crystals have been shown to cause inflammation. As a response to cholesterol crystal accumulation, the NLRP3 inflammasome is activated to produce IL-1β which eventually leads to atherosclerotic lesions. As a part of innate immunity, CARD8 is involved in the modulation of above mentioned inflammatory activities. The primary objective of this study was to investigate the association between polymorphism of CARD8 rs2043211 and susceptibility to arteriosclerosis obliterans (ASO) in Chinese Han male population., Methods: 758 male arteriosclerosis obliterans patients and 793 male controls were genotyped for rs2043211 with the TaqMan allele assays. Fasting blood-glucose (FBG), total cholesterol (TC), triglycerides (TG), urea nitrogen, creatinine, Serum uric acid, high density lipoprotein, low density lipoprotein, ALT, AST, and IL-1β in the blood were detected for all subjects. Clinical data were recorded to analyze the genotype-phenotype. Independent samples t-test was used to perform the comparisons between two groups. Odds ratios (ORs) with 95% confidence intervals (CIs) were calculated to measure the strength of relationship in the genotype distribution and allele frequencies between patients and controls. The analysis of variance was used for a genotype-phenotype analysis of the ASO patients., Results: The genotypic and allelic frequencies in the ASO group were significantly different from that in the control group (P = 0.014 by genotype, P = 0.003 by allele). Those carrying the genotype TT had a higher risk for ASO than those carrying the genotype AA (OR = 1.494, 95%CI1.131-1.974, P = 0.005).The difference was also significant after the adjustment for the history of smoking, TC, LDL, fasting blood glucose, systolic blood pressure and BMI(OR = 1.525, 95%CI1.158-2.009, P = 0.003)., Conclusion: Our finding suggests that the polymorphism of CARD8 rs2043211 is probably associated with the development of ASO in Chinese Han male population., (© 2017 The Author(s) Published by S. Karger AG, Basel.)

The efficacy and prognostic significance of the forced intraarterial injection of medicinal preparations in complex of treatment in patients, suffering chronic ischemia of the lower extremities tissues, was studied. In experimental conditions of the main blood flow blockade the forced intraarterial injection of medicinal preparations have promoted the release of vessels from cellular aggregates and the blood clots; and in the ischemia of degree ІІ - ІІІа – has determined more pronounced clinical improvement. Appearance of reactive hyperemia of the foot in terms more than 2 min have witnessed the hopelessness of the extremity saving.

In the patients, suffering diabetes mellitus type ІІ, treated in 2015 - 2016 yrs for complicated diabetic foot syndrome, a systolic arterial pressure (SAP) on level of the first toe was determined, and roentgenography of the foot in two projections done. The SAP value from 120 to 200 mm Hg and higher have had witness the presence of Menkeberg?s sclerosis stages III - V. Prognostically favorable is a SAP value of 80 mm Hg and higher, and unfavorable data – the SAP value lowering lesser than 80 mm Hg. The SAP value lower than 30 mm Hg have had witness the vessel obliteration and thrombosis occurrence.

Both HIV and antiretroviral therapy could induce vascular aging with unclear mechanisms. In this study, via microarray analysis, we identified, for the first time, that miR-34a expression was significantly increased in both HIV-infected, and antiretroviral agents-treated vessels and vascular endothelial cells (ECs) from these vessels. In cultured ECs, miR-34a expression was significantly increased by HIV-Tat protein and by the antiretroviral agents, lopinavir/ritonavir. Both HIV-Tat protein and antiretroviral agents could induce EC senescence, which was inhibited by miR-34a inhibition. In contrast, EC senescence was exacerbated by miR-34a overexpression. In addition, the vascular ECs isolated from miR-34a knockout mice were resistant to HIV and antiretroviral agents-mediated senescence. In vivo, miR-34a expression in mouse vascular walls and their ECs was increased by antiretroviral therapy and by HIV-1 Tat transgenic approach. miR-34a inhibition could effectively inhibit both HIV-Tat protein and antiretroviral therapy-induced vascular aging in mice. The increased miR-34a was induced via p53, whereas Sirt1 was a downstream target gene of miR-34a in both HIV-Tat protein and antiretroviral agents-treated ECs and vessels. The study has demonstrated that miR-34a is a common link in both HIV and antiretroviral therapy-mediated vascular aging., Competing Interests: The authors declare no conflicts of interest.

In the clinic 60 patients were examined, in whom reconstructive operations on the main arteries were performed for obliterating atherosclerosis (OA) of the lower extremities vessels. Efficacy of impact of Plestazol (in a 200 mg/day dose) on neointima hyperpla- sia was studied. Clopidogrel (75 mg/day) was administered to patients of comparative group. Effective criteria for estimation of the migration-proliferation disorders state in endothelial dysfunction are concentration of the intercellular adhesion molecules (ICAM) and the basic factor of the fibroblasts growth (FGFb); morphological disorders in hyperplastic reactions of neointima - determination of thickness of "intima-media" complex in accordance to the ultrasound duplex scanning data. There was established, that Plestazol constitutes an effective disaggregate preparation, positively impacting decelerating reaction of neointima hyperplasia, including, deceleration of the smooth- muscle cells migration into subendotelial layer in the formation zone of the femoro- popliteal shunt distal anastomosis.

This study was designed to investigate the correlation between autophagy and polarization of macrophages in atherosclerosis (AS) plaque in arteriosclerosis obliterans amputees. Femoral artery specimens from arteriosclerosis obliterans amputees were performed hematoxylin and eosin (HE) staining, oil red O and immunofluorescence staining to observe the morphology of atherosclerotic plaque, phenotype of macrophages and autophagy in plaque; using real-time quantitative RT-PCR technology to detect the mRNA level of M1 and M2 type markers in arterial tissue; to analyze polarized signal pathway and autophagy protein levels in macrophages by Western blotting. Arterial specimens staining showed obvious lipid deposition and obvious infiltration of amount of foam cells and inflammatory cells. Macrophages were mainly expression M1 type in percentage in fibrous plaque. Although both M1 and M2 macrophages were upregulated in atheromatous plaque, the increase was dominant in M2 type in percentage. The level of autophagy was significantly higher in the atheromatous plaque than that of fibrous plaque. The expression of tumor necrosis factor- α (TNF-α), monocyte chemotactic protein-1 (MCP-1), inducible nitric oxide synthase (iNOS), interleukin-6 (IL-6) and interleukin-12 (IL-12) mRNA was significantly higher in fibrous plaque than that of atheromatous plaque (P < 0.01 or 0.05), and arginase-1 (Arg-1), transforming growth factor-β (TGF-β), CD163 and interleukin-10 (IL-10) mRNA was significantly lower than that in atheromatous plaque (P < 0.01). The levels of p-STAT1 and NF-κB were significantly increased in fibrous plaque (P < 0.01), while p-STAT6 expression was significantly increased in atheromatous plaque (P < 0.01). The level of LC3-II was significantly higher in atheromatous plaque than that in fibrous plaque (P < 0.01). Macrophages in early atherosclerotic plaque were induced to M1 type through p-STAT1/NF-κB pathway and expressed moderate levels of autophagy; while macrophages in advanced plaques were induced to polarization of M2 type through p-STAT6 pathway. M2 macrophages expressed a higher level of autophagy than M1 macrophages.

Results of examination of 46 patients, suffering obliterating atherosclerosis of the lower extremities arteries solely or in combination with diabetes mellitus (DM), were analyzed. The malleolar pressure index (MPI), regional systolic pressure (RSP), velocity of the volume blood flow (VVBF), rheographic index (RI), using test with nitroglycerine, postocclusion venous pressure (POVP) and intaosseous pressure (IOP) in tibiae were studied. The RI reduction, parallel to the arterial ischemia progression, was established. The test indices with nitroglycerine in patients with obliterating atherosclerosis have reduced step by step. With coexistent DM the efficacy of nitroglycerine was practically absent. POVP is upgraded in patients of all the groups and it have lowered step by step in a laying position of the patient, and while transition into standing position--it have upgraded progressively with a progress of arterial ischemia. IOP have upgraded significantly in isolated obliterating atherosclerosis in ischemia stage 3a and have lowered--in stage 3b. In coexistent DM IOP is upgraded in ischemia stage 3b also.

Objective: To study the clinical effects of endovascular reconstruction versus bypass surgery for TASC II(trans-atlantic inter-society consensus II) C/D femoropopliteal artery lesion resulted from arteriosclerosis obliterans., Methods: One hundred and three patients(119 limbs)accepted bypass surgery or endovascular therapy for TASCII C/D femoropopliteal artery lesion between January 2002 and December 2012 at our institution were retrospectively assessed.All the patients were diagnosed with arteriosclerosis obliterins, and all their Rutherford classifications were from 2 to 5 degrees.Among them there were 71 limbs treated by endovascular reconstruction and the other 48 limbs were treated with bypass surgery.We evaluated the short term clinical effect according to the condition when patients left the hospital, and evaluated the long term clinical effect according to the results of the patients' latest follow-up in 2014. Their clinical data before treatment, complication rates, death rates, hospital stays, short term and long term effects, reoperation rates, 1 to 10 years primary and secondary accumulative patency rates and limb salvage rates were compared., Results: There was no significant difference between the bypass group and the endovascular group on the mean age and ankle brachial index before treatment [(67.1 ± 7.1) years(51 to 80 years) vs. (68.0 ± 9.4) years (49 to 91 years), P=0.561;(0.41 ± 0.23) vs. (0.40 ± 0.26), P=0.928]. There were more TASCII D patients in the bypass group than those in the endovascular group (P<0.001), and the rutherford classification was higher in the endovascular group than that in the bypass group. The difference in the mean follow-up between the bypass group and the endovascular group was not significant [(41.7 ± 23.6) months vs. (59.5 ± 41.6) months, P=0.065]. Five peri-operative complication cases occurred in the bypass group, including 2 cases of acute thrombosis,1 case of infection and 2 cases of heart failure, and only 1 complication case occurred in the endovascular group that was heart failure.The complication rate was higher in the bypass group than that in the endovascular group [10.4% vs. 1.4%, P=0.039]. And there was no death in both the groups.Compared with the endovascular group, the bypass group had a longer hospital stays [(13.2 ± 4.7) d vs.(6.5 ± 3.1) d, P<0.001], a higher reoperation rate (58.3% vs.31.0%,P=0.003), a better short term, obvious, and effective rate (25.0% vs. 9.9%, P=0.027), a worse long term deterioration rate (37.5% vs. 18.3%, P=0.019) and higher 1 to 10 years primary and secondary accumulative patency rates(P=0.001, P=0.001).There was no significant difference between the two groups on the increase of ankle brachial index [(0.34 ± .28) vs. (0.31 ± 0.23), P=0.371], and short term and long term total effective rates (89.6% vs.84.5%, P=0.426; 45.8% vs. 56.3%, P=0.260), and limb salvage rate (83.3% vs.94.4%, P=0.051)., Conclusion: Endovascular therapy is a safe, effective and minimally invasive therapy for TASCII C/D femoropopliteal artery lesion resulted from arteriosclerosis obliterans.

Aim: RhoA is a critical factor in regulating the proliferation and migration of arterial smooth muscle cells (ASMCs) in patients with arteriosclerosis obliterans (ASO). RhoA is modulated by microRNA-133a (miR-133a) in cardiac myocytes and bronchial smooth muscle cells. However, the relationship between miR-133a and RhoA with respect to the onset of ASO in the lower extremities is uncertain., Methods: We employed in situ hybridization (ISH) and immunohistochemistry (IHC) to detect the location of miR-133a and RhoA in ASO clinical samples, respectively. 5-ethynyl-2'-deoxyuridine (EdU), cell counting kit-8 (CCK-8), Transwell and wound closure assays were utilized to determine the features of human ASMC (HASMC) proliferation and migration. The expression of miR-133a in the HASMCs was assessed using quantitative real-time PCR (qRT-PCR), while that of RhoA was examined via qRT-PCR and Western blotting., Results: We found miR-133a and RhoA to be primarily located in the ASMCs of ASO. miR-133a was significantly downregulated in the ASO tissues and proliferating HASMCs. In contrast, RhoA was upregulated in the ASO samples. The proliferation and migration of HASMCs was markedly promoted by the downregulation of miR-133a and inhibited by the upregulation of miR-133a. The Luciferase assay confirmed that RhoA was a direct target of miR-133a. The upregulation of miR-133a in the HASMCs decreased the RhoA expression at the protein level. Inversely, the downregulation of miR-133a increased the RhoA protein expression. Of note, the overexpression of RhoA in the HASMCs attenuated the anti-proliferative and anti-migratory effects of miR-133a., Conclusions: Our data indicate that miR-133a regulates the functions of HASMCs by targeting RhoA and may be involved in the pathogenesis of ASO. These findings may lead to the development of potential therapeutic targets for ASO of the lower extremities.

Aims: To explore the expression of miR-24-3p in human arteries with arteriosclerosis obliterans (ASO) as well as the role of miR-24-3p in the pathogenesis of ASO., Methods: We used quantitative real-time PCR (qRT-PCR) and in situ hybridization to monitor miR-24-3p expression in human arteries. To investigate the effect of miR-24-3p on human arterial smooth muscle cells (HASMCs), we applied cell counting and EdU assays to monitor proliferation and transwell and wound healing assays to investigate migration and flow cytometry to investigate apoptosis. Furthermore, we applied 3'-untranslated region (3'-UTR) luciferase assays to investigate the role of miR-24-3p in targeting platelet-derived growth factor receptor B (PDGFRB) and c-Myc., Results: MiR-24-3p was mainly located in the media of arteries and was downregulated in ASO arteries compared with normal arteries. Platelet-derived growth factor BB (PDGF-BB) treatment reduced the expression of miR-24-3p in primary cultured HASMCs. MiR-24-3p mimic oligos inhibited the proliferation and migration, and promotes apoptosis of HASMCs. Our 3'-UTR luciferase assays confirmed that PDGFRB and c-Myc were targets of miR-24-3p., Conclusion: The results suggest that miR-24-3p regulates the proliferation and migration of HASMCs by targeting PDGFRB and c-Myc. The PDGF/miR-24-3p/PDGFRB and PDGF/miR-24-3p/c-Myc pathways may play critical roles in the pathogenesis of ASO. These findings highlight the potential for new therapeutic targets for ASO., (© 2015 S. Karger AG, Basel.)

Background: Recent findings have elucidated that netrin-1 has ability of promoting angiogenesis besides the functions in nervous system. Autologous mesenchymal stem cells (MSCs) transplantation is now proved to be an effective method to treat peripheral arterial disease. However there are still many patients who cannot complete full treatments. Therefore it is necessary to improve the effectiveness. This study estimated the curative effects in chronic limb ischemia when MSCs allied with netrin-1., Materials and Methods: Thirty-six rats were made into chronic limb ischemia models. They were randomly assigned to four groups, netrin-1 + MSCs group (treated with netrin-1 and MSCs derived from peripheral blood), MSCs group (treated with MSCs individually), netrin-1 group (treated with netrin-1 individually), and control group (treated with saline). Measurements of murine behaviors, vascular endothelial growth factor expression, and capillary density in ischemia limb were performed on days 7, 14, and 28 after treatments; measurements of contraction force in ischemia limb was performed on day 28 after treatments to compare differences among the groups., Results: Netrin-1 allied with MSCs significantly increased Tarlov score, vascular endothelial growth factor expression, capillary density, and muscular strength in ischemia limb., Conclusions: Netrin-1 allied with MSCs derived from peripheral blood significantly promoted angiogenesis in aged rats with chronic limb ischemia. It may be a promising method of treating peripheral arterial disease in the future., (Copyright © 2014 Elsevier Inc. All rights reserved.)

The causes, influencing possibility of a knee joint preservation, when performing tne lower extremity high amputation, were analyzed in 65 patients while occurrence of purulent-necrotic complications of obliterating angiopathy. In accordance to results of logistic regression the value of comorbidity index 4 and more have appeared significant as a predictor of impossibility to preserve a knee joint. In its bigger or lesser values a success or failure of transtibial amputation is possible.

Results of reconstructive operations, performed on the main arteries of the lower extremities (LE) in 63 patients, suffering obliterating atherosclerosis were analyzed with the objective to analyze the causes of thrombotic complications occurrence. There was established, that common clinical screening tests, applied for control of the hemostasis system state, do not permit completely to reveal thrombophylic changes in early postoperative period timely and to prevent the occurrence of the reconstructed segments thrombosis. Direct correlation connection between the ischemia stage of the LE tissues, the reperfusion changes signs and hyperaggregation severity, occurring due to deficiency of the blood serum natural anticoagulants in patients, suffering obliterating atherosclerosis, was established.

Introduction: There are limited therapeutic options to improve microcirculation., Aim: The question of the study was to investigate any potential beneficial effect of bio-electro-magnetic-regulation therapy on microcirculation in patients suffering from obliterative peripheral arterial disease including the circulation of lower extremities, as well as intermittent claudication., Method: Thirty patients suffering from obliterative peripheral arterial disease (Fontaine IIa and IIb) were recruited. The first step of the study was to determine the pain free and maximal walking distance with a treadmill unit. After the placebo period patients received 8 and 20 minutes bio-electro-magnetic-regulation treatment 16 times. After the treatment the pain free and maximal walking distance were measured again. In the second stage of the study the patients were treated by pentoxifylline infusions., Results: Bio-electro-magnetic-regulation treatment increased the pain free period by 57.4% (p = 0.005) and the maximal walking distance by 36.6% (p = 0.042). The two forms of therapy together increased the pain free and maximal walking distance by 81.9% and by 84.0%, respectively. The combined therapy was very effective in contrast to placebo and bio-electro-magnetic-regulation treatment (p = 0.000373 and p = 0.00741, respectively)., Conclusions: The bio-electro-magnetic-regulation therapy mainly affected the microvessels and pentoxifylline therapy rather had beneficial effects on hemorheology. The clinical effectiveness of combined therapy was good or excellent in 70% of patients.

Background: Immunoglobulin (Ig) G4-related disease has recently been recognized to occur in the cardiovascular system in the aorta and main branching arteries, often manifesting as aneurysms and arteritis/periarteritis. Peripheral arteries (the femoral and popliteal arteries) are frequent sites of arteriosclerosis obliterans (ASO) and occasionally show aneurysms or arteritis. This study re-examined peripheral arterial lesions from the standpoint of IgG4-related disease., Methods: The study comprised 104 patients who underwent surgical treatment of peripheral arterial lesions, including 30 patients with peripheral arterial aneurysms (PAAs) and 74 with ASO. IgG4-related disease was identified on the basis of diffuse infiltration of numerous IgG4-positive plasmacytes as revealed by immunohistochemical examination. Clinicopathologic features were compared between IgG4-related and IgG4-unrelated lesions., Results: IgG4-related disease was found in four of the 30 patients with PAAs (13.3%; two in the deep femoral artery, two in the popliteal artery) but not in any patients with ASO. IgG4-related PAA displayed clinicopathologic features resembling those of other IgG4-related diseases and a characteristic saccular appearance (P = .002)., Conclusions: IgG4-related disease was detected in PAA patients but not in ASO patients. IgG4-related disease thus represents one potential etiology of aneurysm in the peripheral arteries., (Copyright © 2013 Society for Vascular Surgery. All rights reserved.)

Objective: To identify distinct proteins involved in human atherosclerosis obliterans (ASO) by a differential proteomic approach., Methods: Eight atherosclerotic femoral arteries with a mean age of 68.6 years (6 male and 2 female) and 5 normal femoral arteries with a mean age of 44.2 years (3 male and 2 female) were obtained from high amputation patients. Then the first 2-dimensional maps of the proteome of human femoral arteries was plotted to compare ASO and control specimens. Proteomic profiling was to differentiate and identify histological proteins that were associated with ASO. The differentially expressed proteins were sequenced by matrix assisted laser desorption/ionization mass spectrometry (MALDI-TOF-MS). The result was verified by immunohistochemistry (IHC) and Western blot., Results: ASO was associated with distinct patterns of protein expression in the femoral arteries. A total of 25 distinct spots corresponding to 13 different proteins were identified by MALDI-TOF-MS using the NCBI and IPI databases. These proteins were mainly involved in the pathogenetic mechanisms such as inflammation, oxidative stress, proliferation and transformation of SMCs. The low level of heat shock protein 27 (HSP27) in ASO was verified by IHC and western-blot in accord with the result of MS., Conclusion: Proteomic analysis can be used to investigate differentially expressed proteins, which may provide new insights into ASO pathogenesis, such as HSP27.

Results of application of complex treatment of patients with atherosclerosis obliterans of lower limb arteries by means of perfection of an out-patient stage of the surgical help are presented. Rendering assistance in the conditions of surgical branch of city polyclinic makes modern methods of conservative treatment for the overwhelming majority of patients with an obliterating atherosclerosis of arteries of the bottom extremities more accessible, promotes improvement of indicators of safety of extremities and survival rate of patients.

The authors report the results of the application of electrical pulses to the region of lumbar ganglia for the treatment of patients with obliterative atherosclerosis of the blood vessels in the lower extremities. It was shown that the temporary blockade of sympathetic regulation of the lower limb arteries promotes reduction of vascular tone in the distal segments of the lower extremities, stimulates collateral blood flow, and improves microcirculation in ischemic tissues(specifically, following reconstructive surgery on the lower limb arteries).

Background: The architecture of the arterial wall affected with Buerger's disease has been known to be preserved in all three layers, while the one affected by arteriosclerosis obliterans (ASO) is degenerated and destroyed. We analyzed affected arteries with immunohistochemical methods to clarify the differences between Buerger's disease and ASO., Materials and Methods: Crural arteries obtained from 13 patients with Buerger's disease and 6 patients with ASO at our institute were studied. In addition, we examined seven specimens from six patients who were thought to be normal (without Buerger's disease or ASO) as negative control. Immunohistochemical studies were performed on paraffin-embedded tissues. The primary antibodies were urokinase-type plasminogen activator (uPA) and matrix metalloproteinase-3 (MMP-3). Both are known to play an important role of extracellular proteolysis and to activate each other. Additionally, plasminogen activator inhibitor-1(PAI-1) was also examined., Results: In Buerger's disease, PAI-1 was well expressed along the internal elastic lamina. Urokinase-type plasminogen activator and MMP-3 were slightly positive in intima and media. In ASO, a slight amount of PAI-1 was recognized on vessel walls, and both uPA and MMP-3 were strongly positive in media. In addition, in the control group, PAI-1, uPA, and MMP-3 were well expressed in media., Conclusion: In Buerger's disease, PAI-1 was strongly expressed around the internal elastic lamina, while both uPA and MMP-3 were slightly recognized on vessel walls. These findings could be one of the reasons the general architecture of vessel walls in Buerger's disease is preserved., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Taking into account impossibility of reconstruction-restoration operations performance in cases of the distal arterial bed affection, it is necessary to find out and elaborate the methods of indirect revascularization. The cells transplantation performance for stimulation of reparation and angiogenesis in the affected extremity is considered a perspective direction of treatment. Essential stimulation of angiogenesis occurs after transplantation of progenitor stem cells of a fetal liver according to the data, obtained in experiment on laboratory animals. Later, transplantation of the cells was accomplished in patients with chronic ischemia of the extremities. Using method of electron microscopy of endotheliocvtes of the muscular tissue capillaries there was proved, that the cells transplantation causes essential stimulation of angiogenesis in patients, suffering chronic ischemia and constitutes a perspective method of indirect revascularization.

One of perspective trends of treatment in patients, suffering chronic ischemia of extremities, is application of a cellular technologies. This trend needs scientific confirmation, basing on investigations, which discover primary signs, characterizing chronic ischemic impact on the muscular tissue structure and ultrastructure of vascular endothelium in capillaries of the muscle. The results of investigations conducted witness the muscular tissue reconstruction occurrence under impact of chronic ischemia: restoration of muscular fascicles by connective tissue, spoiling young myons regeneration, the capillaries lumen obturation. In addition, in ischemic state the destruction of energetic organelles and endotheliocytes occurs.

Taking into account the impossibility of performance in some situations of reconstructive operative interventions on arteries, it is necessary to look for new methods of indirect revascularization for the extremities ischemia. Adipose tissue constitutes an accessible and sufficient source of multipotent stromal cells (MSC). Experimental investigations were made in a frame of preclinical trial on laboratory animals with the extremity ischemia simulation, and there was proved the essential stimulation of angiogenesis processes after transplantation performance of stromal-vascular fraction of adipose tissue. The work objective was to study the influence of own adipose tissue MSC transplantation on vascular endothelium changes in patients, suffering chronic ischemia of the extremities. Using electron microscopy method there was proved on microstructural level, that in clinical environment the patients, suffering chronic ischemia of the extremities of various etiology, gain undoubted effect of MSC autotransplantation performed with the objective to stimulate the processes of angiogenesis in the ischemic affection region.

The study was aimed at assessing the relationship between the activity (including the inflammatory one) of the atherosclerotic process in the zone ofarterial reconstruction in patients under-going surgery on the aortoiliac arterial segment and long-term thrombotic reocclusions. The study comprised a total offorty-nine patients, with the postoperative follow-up period amounting to 5 years. We carried out morphological studies of the aortic wall in the zone of the proximal anastomosis and the walls of the common femoral arteries in the zone of distal anastomoses. It was determined that the presence of morphometric sign of inflammation and atherosclerotic deformity in the preparations of the aortic and arterial walls was accompanied and followed by increased incidence of long-term thrombotic complications in the reconstruction zone with more pronounced progression of the thrombotic process in the arteries of the peripheral bed distal to the reconstruction site.

In 55 patients, suffering obliterating atherosclerosis and chronic arterial insufficiency, there was accomplished autotransplantation of the bone marrow aspirate (ABMA) because of impossibility of conduction of direct revascularization of the lower extremity in inoperable or operable in question affection of the distal vascular bed, as a monomethod or in simultant fashion with reconstructive operations on proximal segments of vascular bed or using the other methods of indirect revascularization. After the operation in 63.1% of patients it appeared possible to preserve the "lean on" function of the lower extremity in early period and in 52.6%--in the remote period. The ABMA method application had permitted to correct microhemodynamical disorders by stimulation of angiogenesis and development of collateral blood flow in patients, suffering chronic ischemia of lower extremities, especially in the critical ischemia stage.

The present work deals with a comprehensive study of the microbial spectrum of an ischaemic trophic ulcer in gerontological patients with degree IV lower limb chronic ischaemia in order to improve therapeutic outcomes by means of selecting individual antibacterial therapy for a possible prognosis of the results of treatment for an ulcerative-and-necrotic lesion of distal portions of the lower extremities depending on the spectrum of the microflora vegetating in the wound. Based on examining a total of 130 patients aged 70 years and older suffering from lower limb chronic critical ischaemia, we revealed that the pathogen of secondary wound infection isolated in trophic ulcers of the lower extremities influences the total outcomes of surgical management. A conclusion was made on the necessity of a compulsory dynamic scrutiny of the microbial flora of the lower-limb atherosclerotic ulcer, which makes it possible to carefully select individual antibacterial bactericidal therapy with due regard for microbial sensitivity to bactericidal agents.

Objective: To evaluate the clinical effect and restenosis rate of antiplatelet therapy following peripheral artery angioplasty and stenting., Methods: After successful placement of peripheral artery stents to 103 patients with peripheral arterial occlusive disease (PAOD) in were randomized assigned to 2 groups: antiplatelet therapy group receiving clopidogrel 75 mg plus aspirin 100 mg (n = 56) and control group (n = 47) receiving anticoagulation therapy low molecular weight heparin (LWMH) for 7 d plus long-term warfarin. The patients were followed up 1 day, and 1, 6, 12, and 18 months after the operation to undergo color Doppler ultrasonography, and examinations of blood routine, bleeding time, coagulation time, and ankle-brachial Index. The primary endpoint events included major bleeding rate, and composite rate of restenosis and reocclusion. The secondary endpoint events included cardiovascular events, death, and adverse drug reaction., Results: There were no significant differences in the baseline data between these two groups. The thrombotic occlusion rate was 1.8% in the antiplatelet group and 0% in control group, and the restenosis rate was 14.3% in the antiplatelet group and 25.5% in control group (both P > 0.05). The bleeding complication rate of the antiplatelet group was 1.8%, significantly lower than that of the anticoagulation group (19.1%, P < 0.01). There were not significant differences in cardiovascular event rate and mortality 18 months after operation between these two groups., Conclusion: Antiplatelet therapy combined with clopidogrel plus aspirin is effective and safe in preventing restenosis following peripheral artery angioplasty and stenting.

Morphological changes in the arterial wall after reconstructive and endovascular interventions in patients with lower extremity arterial atherosclerosis obliterans are analyzed. The role of intimal hyperplasia in the development of restenosis as a late complication in reconstructive arterial surgery is shown.

Previous studies have suggested that the lower limb arteriosclerosis obliterans (LASO) could be improved by autologous transplantation of either bone marrow mononuclear cells (BM-MNC) or G-CSF-mobilized peripheral blood mononuclear cells (M-PBMNC). However, the number of patients observed was very limited, and little information is available regarding comparison. The present randomised trial was designed to assess which is the better option. One hundred fifty patients with LASO were randomised to either group A (76 cases implanted with M-PBMNC) or group B (74 cases implanted with BM-MNC), and followed up for 12 weeks. Primary outcomes were safety and efficacy of treatment, based on ankle-brachial index (ABI) and rest pain, and analysis was per protocol. Significant improvement of the main clinical index was observed in both groups after transplantation. No transplantation-related complication was observed in either group. Comparative analysis revealed that at 12 weeks after cell implantation, improvement of ABI (difference 0.06 +/- 0.01; p < 0.0001), skin temperature (difference 0.55 +/- 0.25; p = 0.028), and rest pain (difference -0.57 +/- -0.15;p < 0.0001) was significantly better in groupA patients than group B patients. However, there was no significant difference between two groups for pain-free walking distance, transcutaneous oxygen pressure, ulcers, and rate of lower limb amputation. Autologous transplantation of either M-PBMNC or BM-MNC significantly promotes improvement of limb ischaemia in patients with LASO. Comparative analysis indicated that M-PBMNC should be more practical in comparison with BM-MNC in the treatment of LASO.

Background: Severe arteriosclerosis obliterans (ASO) can be intractable, especially in diabetic patients on hemodialysis (HD). Recently, the transplantation of autologous peripheral blood mononuclear cells (PBMNCs) has been reported to have beneficial effects, but the long-term effects and impact on quality of life (QOL) have not been studied., Methods and Results: Autologous PBMNCs were transplanted into 7 diabetic patients on HD who had severe ASO (5 cases with Fontaine IV and 2 with Fontaine III) after administration of 5 microg/kg granulocyte colony stimulating factor; QOL and degree of ischemia was assessed by measuring skin temperature, skin perfusion pressure (SPP), ankle-brachial index (ABI), and ulcer size, and from angiographic findings. At 4 weeks after the procedure, skin temperature was significantly improved, and SPP and ABI also were increased. These beneficial effects persisted for up to 24 weeks. Angiographic findings and ulcer size improved in 3 of 7 and 3 of 4 patients, respectively. SF-36v2 analysis revealed significant improvements in pain scores. No serious complications were detected., Conclusion: Transplantation of PBMNCs resulted in improvement in pain and leg ischemia for over 6 months without serious complications. This therapy is safe and effective for severe ASO in diabetic patients on HD.

Objective: To explore the effects of Tongxinluo capsule (TXL) on the atherosclerosis obliterans (ASO) in iliofemoral artery of rabbits., Method: Rabbits were randomly divided into 7 groups: sham, model, TXL (0.8, 0.4, 0.2 g x kg(-1)), Tongsaimai tablet (0.8 g x kg(-1)) and Laishike (0.002 g x kg(-1)). The animal model of ASO was established with a combined method of mechanical trauma, immunologic injury and high fat fodder feeding. Rabbits were administrated the drugs 8 weeks after surgery. The levels of TC, TG, HDL-C and LDL-C in serum were determined at the time points below: pre-experiment (0 week), pre-drug administration (8 weeks post-surgery), 4 weeks after drug administration (12 weeks post-surgery), 8 weeks after drug administration (16 weeks post-surgery), 12 weeks after drug administration (20 weeks post-surgery). Meanwhile, the behavioral study was performed, the distal skin temperature of the injured hind limb detected. The histopathological changes in iliofemoral artery were examined after opacification., Result: The levels of TC, TG, LDL-C, VLDL-C and TC/HDL-C were decreased significantly in serum of ASO rabbits. The severity of lameness in the injured hind limb was improved. The distal skin temperature was increased. The thickness and the ratio of intima area of the iliofemoral artery of the injured hind limb were decreased, while the stenosis extent was improved., Conclusion: TXL might be beneficial to modulate blood lipid, as well as the prevention and treatment for ASO.

Background: Arteriosclerosis obliterans (ASO) is a serious complication in patients with end-stage renal disease (ESRD) caused by diabetic nephropathy. Adsorption of low-density lipoprotein (LDL) has been performed to treat ASO. While efficacy of this treatment has been reported in limb ischemia, the mechanism underlying the benefit remains unclear. We investigated how LDL adsorption affected soluble adhesion molecules; P-selectin, an endothelial and platelet activation marker; inflammatory cytokines such as interleukin (IL)-1beta, IL-6 and tissue necrosis factor (TNF)-alpha; and lipids in serum., Methods: Selective LDL adsorption by dextran sulfate columns (LDL apheresis) was performed weekly for 10 weeks to treat eight hemodialysis patients with ASO, ESRD, and type 2 diabetes mellitus. Serum was sampled before and immediately after apheresis., Results: LDL apheresis was performed safely. After LDL apheresis lipid concentrations were significantly reduced and clinical findings, such as Fontaine's classification and ankle brachial pressure index values, were improved. Pretreatment concentrations of soluble intercellular and vascular cell adhesion molecules (sICAM-1 and sVCAM-1) and also P-selectin were higher in patients than healthy controls. After apheresis these decreased, especially P-selectin. IL-1beta, IL-6, and TNF-alpha concentrations before apheresis were similar to those in controls and were unaffected by treatment., Conclusion: Effectiveness of LDL apheresis against ASO may involve decreased endothelial cell and platelet activation.

Thrombosis of deep veins of the lower extremities is a serious and frequently occurring disease (with the incidence rate of 160 newly onset cases per 100,000 people annually). A great number of complications are known to develop in case of the ileofemoral thrombosis and a floating pattern of the proximal portion of the thrombus, especially if extending to the vena cava inferior. Appropriate treatment of thromboses of the vena cava inferior should consist in averting dissemination of thrombosis in the proximal direction, preventing pulmonary-artery thromboembolism (PATE), and restoring the patency of the vena cava inferior in order to eliminate the risk of post-thrombotic disease. We present herein our first experience in rheolytic catheter-aided thrombectomy for a floating thrombus of the vena cava inferior in a female patient with an extremely elevated operative risk and pronounced concomitant somatic pathology.

A prolonged state of ischemia of the lower limbs may affect the balance of some metal ion concentrations in blood. Ischemia of a critical degree, surgery and complicated postoperative periods invoke an inflammatory response, increased production of interleukin-6 (IL-6) and acute phase proteins (APhP) as a response to tissue destruction. The aim of the present study was to investigate the modification of Mg, Cu and Zn concentrations in the serum of patients with atherosclerosis obliterans (AO) before surgery and during the postoperative treatment, and the effect of chronic ischemia of the lower limbs on the relationship between the elements. The group studied consisted of 54 men with chronic ischemia of the lower limbs due to AO. The mean value of serum Mg concentration in men with AO was found to be significantly lower, and that of Cu to be higher, in comparison with the control group. In critical ischemia, the mean serum Cu concentration and the ratio Cu/Zn were found higher than in moderate ischemia. The postoperative treatment results in changes in Cu and Zn concentrations in the AO group that are inversely related to the levels before surgery.

Objectives: The aim of the study was to evaluate strength of the dorsal and plantar flexors in patients with obliterating arteriopathy of the lower limbs (stage 2) and to evaluate fatigue., Population and Methods: The comparison was based on two groups of ten patients: one with obliterating arteriopathy, and the second with healthy limbs as a control group. Each group of patients underwent bilateral plantar flexion and dorsiflexion of the ankle on an isokinetic machine, with the following movements: three series of five movements at 120 degrees/s, then three series of three movements at 30 degrees/s in a concentric mode to evaluate strength. The exercise was repeated 20 times at 180 degrees/s to evaluate fatigue. The isokinetic parameters selected were the couple peak and ratio of workload to total weight of the patient. Tests controlled for cardiac frequency, as measured by electrocardiography, and blood pressure., Results: Patients with obliterating arteriopathy of the lower limbs were generally weaker in global strength of plantar and dorsal flexors and had greater muscular fatigue than the control group (P < 0.05)., Conclusion: Decreased performance in the leg muscles in patients with obliterating arteriopathy of the lower limbs is likely due to arterial deficit. Muscular metabolism anomalies such dysfunction of the terminal stages of the oxidative phosphorylation could also affect strength. Use of isokinetics represents an interesting possibility for treatment of patients with multiple cardiovascular abnormalities.

The aim of the present study was to investigate in vitro the differences in P2 receptor mediated responses of human greater saphenous vein (GSV) taken from patients with varicose disease and obliterating atherosclerosis. Samples of the inguinal part of the GSV were taken from the patients who underwent phlebectomia operation due to varicose disease (n=9, VD group) or femoropoplitea bypass operation using auto-vein due to obliterating atherosclerosis of lower extremities (n=11, OA group). The mechanical responses of the isolated segments of GSV to P2 receptor agonists were tested using standard organ-bath technique. ATP (10(-6)-10(-4) M), ADP (10(-6)-10(-4) M) and alpha,betamethyleneATP (10(-8)-10(-5) M) caused concentration-dependent contractions of the veins of both groups, the latter agonist being approximately tenfold more active than first two. ATP at all concentrations tested, alpha,betamethyleneATP at concentrations of 10(-6) and 10(-5) M and ADP at a concentration of 10(-6) M produced significantly higher contractions of the GSV taken from OA group than from VD group. UTP (10(-6)-10(-4) M) caused concentration-dependent contractions of the veins taken from OA group, while in VD group this agonist was virtually without effect. Adenosine (10(-6)-10(-4) M) and 2-methylthio-ATP (10(-7)-10(-5) M) had no significant contractile activity in this tissue in both groups. It is concluded from this study that there are P2 receptor and adrenoceptor mediated contractions in human greater saphenous veins, which are impaired by varicose disease, in contrast to contractions produced by histamine and carbachol which are, if anything, enhanced.