Objective To evaluate the effectiveness and safety of autologous hemopoietic stem cell implantation for peripheral arterial disease (PAD). Methods Randomized controlled trials (RCTs) were identified from CBM (1978 to September 2010), CNKI (1979 to September 2010), MEDLINE (1950 to September 2010), Pubmed (1950 to September 2010), Embase (1970 to September 2010), and Cochrane l ibrary (issue 4, 2010). The papers of the RCTs of cl inical therapeutic studieson PAD treated by autologous hemopoietic stem cell implantation were included and analyzed according to the criteria of the Cochrane handbook. Results Eight RCTs involving 280 patients and 322 extremities were included, with majority of trials of low methodological qual ity. Meta-analysis indicated that autologous hemopoietic stem cell transplantation had an increased ulcer cure rate [RD=0.38, 95% CI= (0.25, 0.50)], a significant improvement in the ankle brachial index [MD=0.11, 95%CI= (0.04, 0.18)], transcutaneous oxygen tension [MD=7.33, 95%CI= (3.14, 11.51)], and pain-free walking distance [SMD=1.35, 95%CI= (0.90, 1.79)], a significant reduction in rest pain scores [MD= —1.70, 95%CI= (—2.15, —1.25)], and a significant benefit in terms of l imb salvage [RD= —0.19, 95%CI= (—0.31, —0.07)]. Only 2 trials reported the side effects of autologous hemopoietic stem cell transplantation, such as l imbs swell ing and concentrations of serum creatine phosphokinase increasing, and the long-term safety was not reported. Conclusion Based on the review, autologous hemopoietic stem cell transplantation may have positive effect on “no-option” patients with PAD. However, the evidence is not b enough due to the general low methodological qual ity, so we can not draw a rel iable conclusion about the effects of autologous stem cell transplantation for PAD at the moment. Further larger, randomized, double bl ind, placebo-controlled, and multicenter trials are needed.
ObjectiveTo study the effect of exogenous insulin on inducing angiogenesis and the expression of vascular endothelial growth factor (VEGF) of hindlimb ischemia of rats with diabetic. MethodsThe hindlimb ischemic model of diabetic rat was established by the ligation of femoral blood vessels of hindlimb in twenty healthy male SD rats and which were divided into model group (n=10) and treatment group (n=10). Another 10 normal rats were selected as control group. Then the expression of VEGF protein and capillary density of muscle tissues of rat hindlimb were detected by Western blot analysis and alkaline phosphatase (APK) stain method, respectively. ResultsThe differences of body weight and blood glucose level of rats before operation and on day 7 after operation were not significant in the control group (Pgt;0.05). The body weight of rat was significantly lower on day 7 after operation than that before operation in the model group (Plt;0.05), while the difference of blood glucose level of rats was not significant in the model group (Pgt;0.05). The body weight and blood glucose level of rat significantly decreased in the treatment group on day 7 after subcutaneous injection of insulin as compared with the level before operation (Plt;0.05). Compared with the control group, the body weight decreased and blood glucose level increased in the model group and treatment group, and the difference was significant (Plt;0.05, Plt;0.01). The body weight of rat in the treatment group was not different from that in the model group (Pgt;0.05), but the blood glucose level of rat on day 7 after operation in the treatment group was significantly lower than that in the model group (Plt;0.05). The relative expression of VEGF protein of muscle tissues of ischemic hindlimbs in the treatment group (155.06±10.26) was significantly higher than that in the model group (94.30±11.23), Plt;0.05, while no expression was found in the control group. The capillary density of muscle tissues of right hindlimb in the control group was significantly higher than that in the model group or treatment group (Plt;0.05), and furthermore, which was higher in the treatment group than that in the model group (Plt;0.05). The difference of capillary density of muscle tissues of left hindlimb was not different among three groups (Pgt;0.05). The capillary density of muscle tissues of right hindlimb was not different from that of left hindlimb in the control group (Pgt;0.05) and which was significantly lower than that of left hindlimb in the model group or treatment group (Plt;0.05). ConclusionInsulin may increase the expression of VEGF protein in ischemic muscle tissue of diabetic rats and protect the ischemic muscle.
ObjectiveTo investigate the significance of Fogarty catheter embolectomy combined with multiple minimally invasive techniques for acute limb ischemia. MethodsClinical data of 88 cases(88 limbs) of lower limb ischemia who were treated in our hospital from Feb. 2007 to Jan. 2011 were collected and analyzed. Of the 88 patients, 46 cases were operated by Fogarty catheter embolectomy(embolectomy group), 42 cases were operated by Fogarty catheter embolectomy combined with multiple minimally invasive techniques(combination group). Comparisons of the clinical indexes of ankle-brachial index(ABI), saturation of blood oxygen(SO2) of toes, and temperature of foot skin before and after operation were performed between the 2 groups, as well as the incidence of mortality, complication, and amputation after operation. ResultsCompared with before operation in the same group, the value of ABI, SO2, and temperature of foot skin in the 2 groups were higher(P<0.05), as well as the 3 kinds of indexes were both higher in combination group after operation(P<0.05). The rates of mortality, amputation, myonephropathic metabolic syndrome(MNMS), osteofascial compartment syndrome, and transient renal insufficiency were 13.04%(6/46), 17.39%(8/46), 26.09% (12/46), 26.09%(12/46), and 13.04%(6/46) respectively, the corresponding rates in combination group were 4.76% (2/42), 7.14%(3/42), 14.29%(6/42), 9.52%(4/42), and 9.52%(4/42) respectively, which were all lower in combination group(P<0.05). ConclusionFogarty catheter embolectomy combined with multiple minimally invasive techniques can be one of the surgical treatment methods for acute limb ischemia, which is characterized by minimally invasive surgery and effective treatment.
ObjectiveTo put forward the concept of acute peroneal artery ischemia syndrome, and to study its typical clinical manifestations and imaging features so as to provide the basis of the evidence-based medicine for the diagnosis and treatment of acute peroneal artery ischemia. MethodsBetween October 2009 and December 2012, 3 cases (2 males and 1 female, aged 57, 68, and 71 years) of acute peroneal artery ischemia syndrome were treated. All the patients displayed typical three symptoms of "peroneal artery blood supply zone pale/red + severe pain of the gastrocnemius muscle + acute drop foot". Medical examination revealed that they all had localized tenderness in the lateral and inferior part of the gastrocnemius muscle, with decreasing of skin temperature. The pulse of dorsalis pedis artery and the posterior tibial artery on affected limbs could be felt. Blood coagulation function and biochemical assay showed that D-dimer, fibrinogen degradation products, creatine kinase, or myoglobin markedly elevated. Magnetic resonance angiography revealed proximal peroneal artery stenosis. All patients were treated with intravenous thrombolysis, anticoagulation, vasodilation, and improving circulation therapy. ResultsThe symptoms of lower limb swelling, pain, and fatigue were improved obviously in all patients after treatment. Blood coagulation function and biochemical assay showed D-dimer, fibrinogen degradation products, creatine kinase, or myoglobin were gradually reduced. The patients were discharged at 9-13 days after treatments, without recurrence during the follow-up of 1year. ConclusionAcute peroneal artery ischemia syndrome is a special type of acute lower limb ischemia, with the three symptoms of "peroneal artery blood supply zone pale/red+ severe pain of the gastrocnemius muscle + acute drop foot" as the main characteristics and should be treated by early active anticoagulant and recanalization therapy.
Objective To explore the mechanism of mesenchymal stem cells （MSCs） transplantation for chronic hindlimb ischemia in Lewis rats by using cell tracer technique. Methods MSCs were isolated and cultured by using density gradient centrifugation and adherence method respectively， then labeled by 5-bromo-2-deoxyuridine （BrdU）. Eight chronic hindlimb ischemia models of Lewis rats were prepared by using suture-occluded method and then divided randomly to MSCs transplantation group and control group， each group enrolled 4 rats， accepting MSCs transplantation and saline respectively. Then on 7 days and 14 days after transplantation， clinical observation， determination of blood flow， and angiography were performed on rats of the 2 groups. At the same time points after previous tests， rats of the 2 groups were sacrificed to get quadriceps tissues and gastrocnemius tissues to perform HE staining and BrdU immunohis-tochemistry. Results The 8 rats were all survived on 14 days after transplantation， with no tumor happened and necroses in the transplanted area. On 14 days after transplantation， the blood flow ratio of operated side to un-operated side in the hindlimb （1.773 vs. 1.279） of rats in MSCs transplantation group and control group increased， and the angiography results showed that there were no much increase in ratio of collateral vessels number （0.908 vs. 0.835）. There were no significant change in the quadriceps tissues and gastrocnemius tissues by HE staining. The BrdU positive kernels located in the inter-stitial substance cells and vascular endothelia cells， and divided differently in different parts of hindlimb at different time points， that the ratio of positive cells in gastrocnemius tissue was higher than those of quadriceps tissue on 7 days after transplantation， but lower on 14 days. Conclusions MSCs transplantation can increases the blood perfusion of hindlimb in the early stage of chronic hindlimb ischemia model， and the possible mechanism may be the paracrine effect of MSCs but not the number increase of collateral vessels.
ObjectiveTo evaluate the dynamic changes of blood flow and blood pressure of acute hindlimb ischemia of rats by laser Doppler flowmetry (LDF) and laser Doppler perfusion imaging (LDPI). MethodsThe acute hindlimb ischemia model of rats was established by resection of rats femoral arteries of left hindlimb. The blood flow and blood pressure between operated and nonoperated hindlimbs were examined by LDF on 2, 7, 14, 28, and 49 d after operation. And the blood flow was evaluated by LDPI on 7 d after operation. ResultsAll rats survived after operation and no hindlimb necrosis occurred. The mean score was 2 on 14 d after operation and 1 on 49 d after operation. The ratio of blood flow between operated and nonoperated hindlimbs on 2 d after operation significantly increased from 1 to 1.31±0.439 (P=0.021). The ratio of blood flow on 7 d (0.82±0.538) and 14 d (0.93±0.294) after operation was significantly lower than that on 2 d after operation (P=0.032 and P=0.019), although the difference between the two former was not significant (P=0.502). Furthermore, the ratio of blood flow on 28 d after operation reached the bottom (0.41±1.970), which was obviously lower than that on 2, 7, and 14 d after operation (P=0.004, P=0.007, and P=0.006). The blood flow of operated hindlimbs recovered approximately the value before operation (0.98±0.093), which was significantly lower than that on 2 d (P=0.010), higher than that on 28 d (P=0.005), but not different from that on 7 d and 14 d after operation (P=0.126 and P=0.382). The ratio of blood pressure between operated and nonoperated hindlimbs on 2 d after operation significantly increased from 1 to 0.47±0.375 (P=0.031). The ratio of blood pressure decreased on 7 d after operation (0.44±0.118), which was not different from that on 2 d after operation (P=0.203). Furthermore, the ratio of blood pressure on 14 d after operation reached the bottom (0.35±0.115), which was obviously lower than that on 2 d and 7 d after operation (P=0.001 and P=0.036). On 28 d after operation, the ratio of blood pressure increased (0.54±0.146), which was significantly higher than that on 14 d after operation (P=0.008), while not different from that on 2 d (P=0.493) and 7 d after operation (P=0.551). The ratio of blood pressure recovered approximately the value before operation (0.97±0.094), which was significantly higher than that on 2, 7, 14, and 28 d (P=0.013, P=0.021, P=0.002, and P=0.031). ConclusionAcute hindlimb ischemia model of rats can be established by resection of rats femoral arteries of left hindlimb and the most serious stage of hindlimb ischemia is on 14-28 d after operation. LDF and LDPI are of importance for monitoring the dynamic changes of rats hindlimb ischemia after operation.
Objective To summarize the research progress of gene-based therapeutic angiogenesis in lower limb ischemia, so as to provide a new method for non-invasive treatment of lower limb ischemia. Method The literatures on studies of gene-based therapeutic angiogenesis in lower limb ischemia in recent years were read and reviewed. Results The incidence of peripheral arterial disease had been increasing annually. How to effectively reduce the amputation rate and mortality rate of patients with critical limb ischemia was still a clinical problem that needs to be solved urgently. A large number of basic and clinical studies had shown that gene-based therapeutic angiogenesis could effectively induce angiogenesis and collateral circulation in ischemic tissue of lower limb, leading to the significant improvements of blood perfusion in ischemic areas. Additionally, the construction of many kinds of new non-viral gene delivery vectors could also improve the safety and effectiveness of gene therapy to a certain extent. Conclusion Although promising therapeutic effect of gene-based therapeutic angiogenesis brings new ideas and strategies for the treatment of lower limb ischemia, issues still exist that have not been solved.
Objective To explore the effective autologous bone marrow stem cell dosage for treatment of severe lower limb ischemia. Methods From December 2003 to December 2004, 22 cases of bilateral lower limb ischemia were treated with autologous bone morrow cell transplantation. All the patients were randomly divided into two groups according to ischemia degree. In group A(severe ischemia side), the amount of transplanted autologous bone marrow cells was more than 1×108, and ingroup B(mild ischemia side), the amount was less than 1×105. A series of subjective indexes, such as improvement of pain, cold sensation and numbness, and objective indexes, such as increase of ankle/brachial index (ABI) and transcutaneous oxygen pressure (TcPO2), angiography, amputation rate, and improvement of foot wound healing were used to evaluate the effect of autologous bone marrow stem cells implantation. Results The rates of pain relief were 90.0% in group A and 16.7% in group B (Plt;0.01); the rates of cold sensation relief were 90.5% in group A and 5.3% in group B（Plt;0.01）;the improvement of numbness was 62.5% in group A and 9.1% in group B（Plt;0.01）. Increase of ABI was 31.8% and 0 in groups A and B respectively（Plt;0.01） at 4 weeks after implantation. Increase of TcPO2was 94.4% and 11.1% in groups A and B respectively（Plt;0.01） at 4 weeks after implantation. Twelve cases of angiography showed rich new collateral vessels in 100% of the limbs in group A while no remarkable new collateral vessel in group B. The amputation rates were 4.5% in group A and 27.3% in group B（Plt;0.05） at 4 weeks after implantation. The rate of improvement of foot wound healing was 75% in group A and there was no changein wound healing in group B after 4 weeks of implantation. Conclusion The effectiveness of autologous bone marrow stem cell implantation depends on the number of implanted stem cells. Effectiveness is expected in most patients if the implanted stem cell is more than 1×108, whereas there would be little effect if the cell number is less than 1×105.
Objective To evaluate the effects of Shengji Yuhong collagen on promoting angiogenesis of the ischemia tissues and probe the possible mechanisms. Methods Forty-eight Wistar rats were divided by random method of paired into blank group， control group （collagen），and experimental group （Shengjiyuhong collagen）. After made the rats hind limb ischemia model， collagens with or without the extracts of Shengji Yuhong Gao were randomly paired implanted locally in hind limb ischemia tissues of rats in experimental group or control group. The samples of collagens and tissues about 0.5 cm large surrounding the collagen were explanted respectively on day 3，7， 14， and 28 for detected the hemog-lobin contents in colagen， microvascular counting by using CD34 immunohistochemical markers， and the expressions of HIF-1α mRNA and VEGF mRNA by using real-time fluorescent quantitative RT-PCR. The blood perfusion of the ischemic tissues at each time were determined by using laser speckle imaging system of Moor-FLPI. Results The results of Moor-FLPI showed that the obvious ischemia condition after model made， the blood perfusion was significantly lower than that before operation （P＜0.01）. On day 3 after operation it showed obvious congestion in the ischemic tissues， and from day 7 to day 14， it showed the ischemia state locally till day 28 after operation which showed improved situation of ischemic. Except for the day 3， the blood perfusion of experimental group were higher than those of blank group （P＜0.05）. There was no statistical significance between the blank group and control group （P＞0.05）. The blood perfusion on day 7 and day 14 after operation of experimental group were higher than those of control group （P＜0.05）. The hemoglobin contentsof each time point in the experimental group were higher than those in the control group （P＜0.01）. The microvascular counting on day 7 and day 14 in experimental group were higher than those of control group （P＜0.05）. The expressions of HIF-1α mRNA and VEGF mRNA at each time point of experimental group were higher than those of control group and blank group （P＜0.05）， and there was no significant differences between the control group and blank group （P＞0.05）. Conclusion The effects on promoting angiogenesis of rat hind limb ischemia tissues with Shengji Yuhong collagen may though inducing the expressions of HIF-1 α mRNA and VEGF mRNA locally.
Objective To establish chronic hindlimb ischemia model with suture-occluded method in rats， and then compare the effects of chronic hindlimb ischemia model with acute ischemia model. Methods Models of chronic hindlimb ischemia were established by using suture-occluded femoral artery method. The laser Doppler blood flow analysis and angiography were performed on day 7， 14， 28， 42， and 49 after operation， and then the rats were sacrificed after angiography， respectively， the quadriceps and gastrocnemius of contralateral and ipsilateral （surgical side） were gotten， which were tested by HE staining and α-actin immunohistochemistry staining， and then calculate arteriolar density. Results There were no lameness and limb necrosis after operation in chronic hindlimb ischemia models. Laser Doppler analysis found that chronic hindlimb ischemia models were still maintained in ischemia state on day 49 after operation compared with acute ischemic models. The resluts of HE staining showed no acute necrosis and muscle fibrosis in chronic hindlimb ischemia model group. Chronic hindlimb ischemia models after operation did not appear obvious lameness and limb necrosis. The arteriolar density of quadriceps femoris on day 7 after operation in chronic hindlimb ischemia models were less than that in acute hindlimb ischemia models （0.015 2 vs. 0.036 4）. Conclusions Compared with the commonly used acute ischemic models， the duration of arterial limb ischemia in chronic hindlimb ischemia rats， which were established by suture-occluded method， is longer and less likely to be affected by the compensatory mechanisms. So suture-occluded method can provide a new animal hindlimb ischemia model for further study of ischemia angiogenesis mechanism and treatment of severe lower extremity ischemia.