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.
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 compare the effectiveness of autologous implantation between bone marrow stem cells and peripheral blood stem cells for treatment of lower limb ischemia. Methods From December 2004 to December 2005, 42 patients with unilateral lower limb ischemia were treated with both autologous bone marrow stem cell implantation(group A, n=21)and autologous peripheral blood stem cell implantation (group B, n=21). Fortytwo patients included 32 males and 10 females. The age ranged from 34 to 80 years, with a mean of 65.6 years. Of the patients, there were 28, 8 and 6 patients suffered from diabetic lower limb ischemia, Burger’s disese and atherosclotic occlusion, respectively. Ischemic history was from 3 months to 5 years, with amean of 2.1 years. 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), transcutaneous oxygen pressure (TcPO2), angiography, amputation rate, and improvement of foot wound healing, were used to evaluate the effect. Results After 4 weeks of implantation, the rate of pain relief was 88.2% in group A and 89.5% in group B (Pgt;0.05) ; the rate of cold sensation relief was 94.4% in group A and 94.7% in group B（Pgt;0.05）; improvement of numbness was 69.2% and 66.7% respectively in groups A and B（Pgt;0.05）. Increaseof ABI was 38.1% in group A and 33.3% in group B（Pgt;0.05）; increase of TcPO2 was 85.7% and 90.5% respectively in groups A and B（Pgt;0.05）; angiography was performed in 12 patients (group A) and 9patients (group B), and the new formed collateral vessel rate was 83.3% in group A and 77.8% in group B（Pgt;0.05）; the amputation rate was 9.1% in groups A and B（Pgt;0.05）; the rate of improvement of foot wound healing was 60.0% in group A and 66.7% in group B(P＞0.05). Forty patients were followed up 3-15 months(mean 8 months). The improvement rate of subjective symptoms was 75.0% in group A and 70.0% in group B (Pgt;0.05); increase of ABI was 60.0% in group A and 65.0% in group B; increase of TcPO2 was 80.0% and 75.0% respectively in groups A and B; the new formed collateral vessel rate was 90.0% in group A and 84.6% in group B. All ulcers healed except 1 case in group B. Conclusion Bone marrow stem cell graft and peripheral blood stem cell graft are all effective in treatingower limb ischemia.
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.
Objective
To evaluate effect of hypoxia condition (1% or 5% oxygen concentration) on proliferation, adhesion, migration, or viability ability of bone morrow-derived endothelial progenitor cells (EPCs).
Methods
The bone marrow mononuclear cells of SD rat were acquired with density gradient centrifugation method. They were cultured, induced, and differentiated to the EPCs. Then they were cultured respectively in three different oxygen concentrations (1%, 5%, or 21%). On the 3rd day and the 7th day, the effects of the different oxygen concentrations (1%, 5%, or 21%) on the EPCs’ neovascularization characteristics (including proliferation, adhesion, migration, and viability abilities) were evaluated.
Results
Whether cultured for the 3rd day or 7th day, the proliferation, adhesion, migration, and viability abilities of the cultured cells in the 1% and 5% oxygen concentrations were significantly better than those of the cultured cells in the 21% oxygen concentration (all P<0.05). Except for the proliferation ability of the cultured cells in the 5% oxygen concentration was significantly better than that of the cultured cells in the 1% oxygen concentration (P<0.05) on the 3rd day, and the adhesion ability on the 3rd day and the proliferation ability on the 7th day had no significantly differences, the other abilities (adhesion, migration, and viability abilities) of the cultured cells in the 1% oxygen concentration were significantly better than those of the cultured cells in the 5% oxygen concentration (allP<0.05).
Conclusion
Different oxygen concentration has an effect on proliferation, adhesion, migration, or viability ability of bone morrow-derived EPCs, appropriate hypoxia condition (1% or 5% oxygen concentration ) can enhance these abilities.
ObjectiveTo investigate the protective effect of Shenfu injection on liver injury in rats with hind limb ischemia-reperfusion and its mechanism.
MethodsSixty-four male rats were randomly divided into sham operation group, ischemia-reperfusion group, Shenfu group〔Shenfu injection 7.5 mL/kg injection of peritoneal(ip), given 10 min before ischemia-reperfusion〕, Shenfu+Znpp group(Shenfu injection 7.5 mL/kg+Znpp 5 mg/kg ip, given 10 min before ischemia-reperfusion), 16 rats in each group. The rat model of hind limb ischemia-reperfusion injury was reproduced by occluding the hind limb artery of the rats for 3 h and subsequent reperfusing for 4 h. The liver tissues were gathered for malondialdehyde(MDA)and superoxide dismutase(SOD)determination. The expression of hemeoxygenase 1(HO-1)protein in the liver tissues was detected by immunohistochemistry. The pathological changes of liver were observed under the light microscope. The changes of serum glutamate-pyruvate transaminase(GPT)and glutamine oxaloacetic transaminase(GOT)were observed respectively.
Results①Compared with the sham operation group, the contents of MDA, GPT, GOT, and the expression of HO-1 protein were markedly increased in the ischemia-reperfusion group, Shenfu group, and Shenfu+Znpp group(P < 0.05), the activities of SOD were markedly decreased in the ischemia-reperfusion group and Shenfu+Znpp group(P < 0.05).②Compared with the ischemia-reperfusion group, the contents of MDA, serum GPT, GOT, and the expression of HO-1 protein were markedly decreased, the activity of SOD was markedly increased in the Shenfu group(P < 0.05).③Compared with the Shenfu group, the contents of MDA, GPT, GOT were markedly increased, the activity of SOD was markedly decreased in the Shenfu+Znpp group(P < 0.05). Unde ther light microscope, the pathological changes induced by ischemia-reperfusion were significantly attenuated by the Shenfu injection in the Shenfu group and were reversed by the Znpp in the Shenfu+Znpp group.
ConclusionShenfu injection inhibits liver tissue injury during hind limb ischemia-reperfusion, this protective effect might be partly through induction of HO-1.
ObjectiveTo evaluate the value and experience of the multidisciplinary team (MDT) approach in the management of patients with lower extremity arteriosclerosis obliterans (ASO). MethodsA retrospective analysis was conducted on 46 consecutive patients with lower extremity ASO who were treated with MDT model at Zhongshan Hospital, Fudan University, from May 2021 to April 2024. All subjects had critical limb ischemia (Rutherford category ≥4) with comorbidities involving two or more organ systems. Overall mortality, above-ankle amputation rate, and below-ankle amputation rate were recorded. The frequency and depth of involvement of each specialty in the MDT process were also documented. ResultsOf the 46 patients, 37 were male and 9 were female, with a age of (74.3±11.8) years. Major comorbidities included heart disease, hypertension, cerebrovascular disease, diabetes, chronic kidney disease, hyperlipidemia, and others. Overall mortality was 13.0% (6/46). The total amputation rate was 32.6% (15/46), comprising above-ankle amputation in 19.6% (9/46) and below-ankle amputation in 13.0% (6/46). Fourteen disciplines participated in the MDT; in addition to vascular surgery, the most actively involved departments were endocrinology, cardiology, and nephrology. ConclusionsThe MDT model offers unique advantages in the management of critical lower-extremity ASO. By coordinating revascularization timing, extent, and modality, prioritizing comorbid conditions, tailoring operative plans, and optimizing perioperative support, the MDT approach reduces mortality, improves limb salvage rate, and enhances both prognosis and quality of life.
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.
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.
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.
