Objective To study the effect of vein-occlusion on the replanted limb survival in SD rats at different stages. Methods Twenty-five adultSD rats were randomly divided into 5 groups according to the time of the femoral vein occlusion after the replanted limbs:2- ,3- ，4 -，6-，and 8- day groups. The limbs were observed through naked eye, measurement of dermal temperature and angiography. Results No formation of collateral veinlet was found, and necrosis wasseen in the replanted limbs of 2- , 3- day groups. Reflux-vein was gradually increased in the replanted limbs of 4，6，and 8 day groups. Angiographic score of capillary density and dermal temperaturein the thigh muscles were greater in groups 4-，6-，and 8- day than in groups 2 and 3 day. Conclusion Within 2 and 3 days，the replanted limbs of SD rats will necrose because of vein-occlusion; and 4 days later the replanted limbs can survive depending on the reflux-vein of new collateral veinlet.
Objective To investigate the effect and mechanism of epigallocatechin-3-gallate (EGCG) on restenosis of the vein graft. Methods Totally 90 Sprague-Dawley rats were randomly divided a the control group, a vein graft group and an EGCG+vein graft group. At week 1, 2 and 4, the intimal and tunica thickness of the venous graft wall was evaluated by hematoxylin-eosin staining, and the expression of Ki-67 was assessed by immunohistochemistry analysis, and then the expression of hairy and enhancer of split-1 (HES1) was measured by Western blot assay. Results At week 2, the intimal thickness (46.76±4.89 μmvs. 8.93±0.82 μm, 46.76±4.89 μmvs. 34.24±3.57 μm), tunica thickness (47.28±4.37vs. 16.33±1.52 μm, 47.28±4.37vs. 36.27±3.29 μm), positive cell rate of Ki-67 (21.59%±2.29%vs. 1.12%±0.22%, 21.59%±2.29%vs. 15.38%±1.30%), expression of HES1 respectively increased in the experimental group than those in the control group and the EGCG+vein graft group (P<0.05, respectively). At week 4, the intimal thickness (66.38±6.23 μmvs. 8.29±0.79 μm, 66.38±6.23 μmvs. 48.39±4.23 μm), tunica thickness (63.27±6.18 μmvs. 15.29±1.49 μm, 63.27±6.18 μmvs. 44.63±4.49 μm), positive cell rate of Ki-67 (33.19%±3.03%vs. 1.09%±0.19%, 33.19%±3.03%vs. 24.37%±2.73%), expression of HES1 increased in the experimental group than those in the control group and EGCG+vein graft group (P<0.05, respectively). Conclusion EGCG may inhibite restenosis of vein graft by inhibiting Notch signal pathway.
Objective To construct vectors that express phosphatidylinositol-3-kinase, catalytic, beta polypeptide （PIK3cb） shRNA in eukaryon plasmid catalyzed by PI3K in rat, then test their effects on intimal hyperplasia in transplanted vein graft. Methods One hundred and fifty SD rats were randomly divided into six groups （n=25, in each group）: blank (25% Pluronic F-127), shRNA-1, shRNA-2, 1/2 （shRNA-1+shRNA-2）, negative control (pGenesil-1 scramble shRNA) and positive control （wortmannin） group. The jugular vein in rats were interpositioned autologously into the common carotid artery. shRNA and 25% Pluronic F-127 were mixed and coated around the transplanted vein in three PIK3cb shRNA groups. Every 5 samples were removed according to the time point (1, 3, 7, 14 and 28 days after operation), respectively. The thickness of intima and neointima area were calculated and analyzed by computer system. The PCNA expression was detected by Western blot and SP immunohistochemistry. Results The intimal thickness of three PIK3cb shRNA groups were lower than those in the blank group and negative control group on day 3, 7, 14, 28 after operation (P＜0.05); The neointima area in three PIK3cb shRNA groups (except shRNA-2 group on day 3, 7) began to decrease significantly from day one (P＜0.05). The protein expression of PCNA in three PIK3cb shRNA groups on day 3 after operation were decreased compared with blank group and negative group (P＜0.05). The percentage of the PCNA positive cells area in three PIK3cb shRNA groups were significantly lower than those in blank group and negative control group in each time point (Plt;0.05). There were no significant differences between blank and negative control group in different time points (Pgt;0.05). Conclusion The PIK3cb shRNA can effectively inhibit the proliferation of vascular smooth muscle cell, which may provide a new gene therapy for the prevention of vein graft restenosis after bypass grafting.
Objective To study the mechanism of restenosis of the vein graft and the effect of the grafting injury to the vein graft. Methods One side of the 36 healthy rabbits was randomly chosen as the V-A group, and on the side a 1.5cmlong femoral vein was obtained, and an 0.5-cm-long segment of the obtained femoral vein was separated as the control group. The remaining 1-cm-long femoral vein was inverted and was autogenously implanted into the femoral artery on the same side of the rabbit. The other side of the rabbits was chosen as the V-V group, and on this side a 1-cm-long femoral vein was obtained ex vivo and then was sutured in situ. The vein grafts on both sides were harvested 4 weeks after operation. The specimens from the harvested vein grafts were stained with HE and theelastic fiber Victoria blue for an observation on the histological changes in the walls of the vein grafts, and the specimens were also stained by the immunohistochemistry of the proliferating cell nuclear antigen (PCNA) for an observation on the wall cell proliferation of the vein grafts. The changes in the ultrastructure of the proliferated wall cells of the vein grafts were observed under electron microscope. The two sides of the rabbits were compared. Results The smooth muscle cells of the media developed hyperplasia, but theintima and the media remained unchanged in their thickness (3.50±0.41 μm, 12.23±1.59 μm) in the V-V group, with no difference when compared with the control group (3.40±0.37 μm, 12.14±1.62 μm); however, when compared with the V-A group (25.60±3.21 μm, 21.30±2.47 μm),there was a significant difference in the thickness (Plt;0.01). There were no cells positive for PCNA by the immunohistochemistry examination in the control group. The cells positive for PCNA were found in the intima and the media in both the V-V group and the V-A group; however, the percentageof the cells positive for PCNA in the intima and the media was significantly greater in the V-A group than in the V-V group (16.4%±1.9% and 36.5%±3.7% vs 5.9%±1.3% and 23.4%±3.4%, Plt;0.01). In the V-V group, the endothelial cell could be observed under transmis-sion electron microscope, which was flat and had a processlike villus at its free end, and the endothelial cells were closely arranged andhad hyperplasia of the smooth muscle cells in the media. But in the V-A group,the endothelial cells had an obvious hyperplasia with an irregular shape and a widened space between the cells, and in the intima a great amount of the smooth muscle cells could be observed, which had a broken basement membrane. The smooth muscle cells also had an obvious hyperplasia in the media. The shape and alignment of the endothelial cells in the control group were similar to those in the V-V group, but the hyperplasia of the smooth muscle cells was not observed in the media. Conclusion The grafting injury can cause hyperplasia ofthe vascular wall cells, and if the hemodynamics is changed simultaneously, more serious hyperplasia and cell migration can be observed from the media to the intima, resultingin restenosis of the blood vessels. So, if we can reduce the grafting injury and improve the microcirculation of the vein graft, we may find out the methods ofpreventing restenosis of the vein graft. The animal model of the V-V graftcan help to understand the mechanism of restenosis of the vein graft.
【Abstract】ObjectiveSome studies have demonstrated that recombinant adenoviruses are efficient vectors for gene transfer to the venous wall and AdCMV.tk encoded thymidine kinase can be used to reduce restenosis. In this study AdCMV.tk was apply to human vein smooth muscle cells (SMC) and organ cultured saphenous veins to study its effects on proliferation of SMCs and reduction of intimal hyperplasia. MethodsThe adenovirus vector transferred tk gene and mark gene lacZ to the SMC of human saphenous veins and organ cultured vein segments. Various concentrations ganciclovir (GCV) were contained in culture media. The efficiency of gene transfer was studied by using Xgal staining. The proliferation of SMC was monitored by the method of trypan blue exclusion. The bystander effect was observed by mixed cell culture. After vein segments treated by AdCMV.tk+GCV and cultured for 14 days, HE and VG staining were carried out and intimal thickness was analysis by computer image system. ResultsAdenovirus vector could infect saphenous vein SMC efficiently both in cultured SMCs and organ cultured vein segments. Gene expression sustained 14 d at least. The inhibition of SMCs proliferation in vitro was a positive correlation in GCV concentrations and the levels of tk expression. The proliferation of SMCs transfectered lacZ wasn’t restrained by GCV (P＜0.05). In mixed cell experiment there was at least 55% reduction in total cell number when as few as 10% of the cells express tk. Assessment of this “suicide gene strategy” in saphenous vein organ culture model demonstrated that veins treated with AdCMV.tk+GCV had a significant reduction at 14 days in the intimal thickness compared to control group (P＜0.01). ConclusionThe results suggest that adenovirusmediated gene transfer of tk along with GCV administration may be a useful strategy to treat the proliferation of intimal hyperplasia of transplanting saphenous veins. Bystander effects are amplified by AdCMV.tk/GCV gene therapy system.
ObjectiveTo investigate the expression of extracellular signalregulated kinase (ERK) and p38 mitogenactivated protein kinase （p38 MAPK） in autogenous vein grafts during vascular remodeling.MethodsAn autogenous vein graft model was established by transplanting the right jugular vein to infrarenal abdominal aorta in 80 Wistar rats. Vein graft samples were harvested 6 hours, 24 hours, 3 days, 7 days, 2 weeks, 4 weeks, 6 weeks and 8 weeks after surgery. Gene expression of ERK and p38 MAPK was measured by reverse transcriptionPCR. Western blot was used to detect the expression of protein products and phosphorylation protein products of ERK and p38 MAPK. Apoptosis of vascular smooth muscle cells (VSMCs) was determined by TUNEL. Proliferating cell nuclear antigen(PCNA) of VSMCs also was studied.ResultsThe expression of ERK1 mRNA and p38 MAPK mRNA increased considerably after surgery. ERK1 mRNA reached the peak on the 7th day 〔（33.2±14.2）%, P＜0.01〕, but p38 MAPK mRNA reached the peak on the second week after surgery 〔(58.8±26.2）%, P＜0.01〕. The expression of ERK1/2 detected by western blot reached the peak during 1 to 2 weeks and decreased gradually to normal level 6 weeks after surgery. The expression of p38 MAPK reached the peak during 2 to 4 weeks and decreased to 1/4 to 1/2fold 8 weeks after surgery. There was a positive relationship between ERK1 and PCNA(r=0.759 6，P＜0.01) and a positive relationship between p38 MAPK and apoptosis(r=0.892 2，P＜0.01). ConclusionActivation of MAPK system exists in autogenous vein grafts and it may become a new target for the therapy of stenosis after vein grafts.
Objective To assess the curative effect of percutem transilluminated with negative pressured on the potaried technique on the treatment of venous ulcer in lower extremity. Methods The clinical date of 300 cases involving 300 legs with venous ulcer in lower extremity， who underwent the percutum transilluminated negative pressured potaried technique using TRIVEXTM Ⅱ potaried system or the percutum transfixion surgical treatment from October 2005 to June 2009, were analyzed. Three hundred cases were randomly divided into potaried group and transfixion group. In potaried group, there were 190 cases involving 190 legs treated with TRIVEXTM Ⅱ potaried system. In transfixion group, 110 cases involving 110 legs treated with percutum transfixion. The clinical indexes of skin infection rate and skin necrosis rate, shrinkage rate of wound area and skin depigmentation rate, ulcer healing rate and ulcer recurrence rate were calculated to assess the clinical curative effect on day 5, day 20, day 120 and day 360 after operation respectively. Results The rates of skin infection and skin necrosis were significantly decreased in potaried group compared with transfixion group on day 5 after operation (P＜0.05), the rates of shrinkage of wound area and skin depigmentation were significantly increased in potaried group compared with transfixion group on day 20 (P＜0.05). The ulcer healing rate was not significantly different between the two groups on day 120 (Pgt;0.05). Ulcer recurrence rate was remarkably lower in potaried group than that in transfixion group on day 360 (P＜0.05). Conclusion It can be concluded that percutem transilluminated with negatived pressured on the potaried technique with TRIVEXTM Ⅱ potaried system can efficiently promote the healing of venous ulcer in the lower extremity, and at the same time it has an ascendancy in lessening skin infection and skin reinjury.
Objective To investigate the procedure and clinical effect of revascularization for arterial occlusion in lower extremity. Methods From July 1998 to March 2005, 29 cases of arterial occlusion were treated by microsurgery. Of 29 cases, there 22 males and 7 females, aging 22-86 years, including 9 cases of thromboangiitis obliterans(TAO), 17 cases of arterial sclerosis obstruction(ASO) and 3 cases of diabetic foot(DF). The location was the left in 17 cases, the right in 11 cases and both sides in 1 case. All cases were inspected by color-Doppler ultrasonic scanning before operation. The cases of ASO and DF were checked with MRA. The results of examinations showed that the locations of arteriostenosis and obstruction were: in 9 cases of TAO, the distal superficial femoral artery in 3 cases, popliteal artery in 5 cases, bilateral dorsal metatarsal artery in 1 case; in 17 cases of ASO, common iliac artery in 2 cases, external iliac artery in 4 cases, femoral artery in 10 cases and popliteal artery in 1 case; and were all superficial femoral artery in 3 cases of DF. DSA examination confirmed that there was appropriate outflow in 15 cases. Basing on the location and extent of the arterial occlusion, 11 cases were treated by the primary deep vein arterializing, 16 cases by arterial bypass distribution and 2 cases of extensive common iliac arterial occlusion were amputated in the level of 1/3 distal thigh. Results The postoperative duration of follow-up for all cases was 3 months to 7 years. In 9 cases of TAO, 2 healed by first intention after deterioration, 4 healed after changing dressing and 3 had fresh soft tissue growth after debrided superficial secondary necrosis. In 17 cases of ASO, 13 healed by first intention, 2 healed after changing dressing and 2 were amputated. In 3 cases of DF, 2 healed after changed dressing and debrided, 1 was aggravated with the second toe necrosis. Conclusion Performing primary deep veinarteriolization and arterial bypassdistribution is effective for treatment of arterial occlusion of lower extremity. The arterial reconstructive patency rate can be improved by microsurgical treatment.
The experiment was earied out on the a boomen of the whiterats. The epigastric vein wasarterialized by means of anastomcois with the femoral artery, lateral thoracic vein was reserved as aefferent vessel. The changes of hemorheology were mesured after arterialization, and were comparedwith the changes in the normal A-V skin flaps. The levels of platelet, aggreation, blood viscosityand plasma fibrinogen in arterialized vein flape were signmeantly higher than that in A-V flaps. ASa r...
In order to prevent tendon adhesion following operation, autogenous great saphenous vein graft was used to reconstruct the tendon sheath. The operation was performed under microsurgical technique. This method was used to repair 23 tendons and 17 tendon sheaths. The early functional exercises were carried out after operation. Follow up from 10 months to 4 years, the prognosis was good except in 3 fingers, in which, the wounds were infected resulting the necrosis of the grafted veins and exposure of the repaired tendons. The details of the operation were introduced. It was emphasized that non-traumatic handling of the tissues was essential in preventing tendon from adhesion.