Role of Vascular Endothelial Growth Factor-C during Stem Cell Therapy Using Autologous Bone Marrow Mononuclear Cells in Patients with Lower Limb Lymphedema

Introduction: Vascular endothelial growth factor-C (VEGF-C) is the primary lymphangiogenic factor that stimulates lymphangiogenesis by signaling via specific receptor, vascular endothelial growth factor receptor 3 (VEGFR3). This study was conducted to evaluate the change in the level of VEGF-C before and after autologous bone marrow mononuclear cell transplantation for treatment of Lower limb lymphedema. Patient and methods: Forty patients with lower limb lymphedema were divided into two groups. Group I included 20 patients with chronic lower limb lymphedema who underwent autologous bone marrow mononuclear cell transplantation. Group II included 20 patients with chronic lower limb lymphedema who were exposed only to compression therapy as a control group. VEGF-C level in the diseased limbs was measured in both groups at the beginning of the study then 3 and 6 months respectively. Results: Group I included 20 patients, 8 patients were male (40%) and 12 patients were females (60%) with mean age 29.5 ± 12.15 while group II included 20, 10 patients were male (50%) and 10 patients were females (50%) with mean age 39.5 ± 11.5. In group I, the specimens were taken at 3 and 6 months after transplantation showed a marked decrease in the VEGF-C level with statistically significant p value, 0.02 and 0.001 respectively. In group II the level of VEGF-C after compression therapy alone at 3 and 6 months interval showed fluctuation with statistically non-significant p value, 0.64 and 0.55 respectively. Conclusion: VEGF-C is essential for regulation of lymphangiogenesis. The level of VEGF-C was found elevated in patients with lymphedema and decrease after autologous mononuclear bone marrow cells, however these results were statically non-significant.

Influence of Transplantation of Allogenic Bone Marrow Mononuclear Cells on the Left Ventricular Remodeling of Rat after Acute Myocardial Infarction

To probe into the influence of transplantation of allogenic bone marrow mononuclear cells （BM-MNCs） on the left ventricular remodeling of rat after acute myocardial infarction （AMI）, 60 male Wistar rats were evenly divided into three groups at random： control group 1, control group 2 and transplantation group. In control group 1, chest was opened without ligation of coronary artery; in control group 2 and transplantation group, the left anterior descending branch of coronary artery was ligated to establish AMI model. Prepared culture medium and allogenic BM-MNCs suspension were respectively implanted the surrounding area of infracted cardiac muscle via epicardium of control group 2 and transplantation group. Four weeks after the operation, the osteopontin gene （OPN mRNA, P〈0.01）, type Ⅰ collagen （P〈0.01） and angiotensin Ⅱ （AngⅡ, P〈0.01） content in the left ventricular non-infracted myocardium, and the Ang Ⅱ density in blood plasma （P〈0.05） of transplantation group and control group 2 were all significantly higher than that of control group Ⅰ. In the transplantation group, the myocardial OPN InRNA, type Ⅰ collagen and Ang Ⅱ content of non-infracted zone in left ventricle, and the Ang Ⅱ concentration in blood plasma were all significantly lower than those of control group 2 （P〈0.05 for all）. It is concluded that allogenic BM-MNCs transplantation may ease left ventricular remodeling after AMI by inhibiting the synthesis of type Ⅰ collagen in the cardiac muscle and down-regulating the expression of Ang Ⅱ and OPN gene.

Effect of Intracoronary Infusion of Bone Marrow Mononuclear Cells or Peripheral Endothelial Progenitor Cells on Myocardial Ischemia-reperfusion Injury in Mini-swine

Objective To simulate and assess the clinical effect of intracoronary infusion of bone marrow mononuclear cells or peripheral endothelial progenitor cells on myocardial reperfusion injury in mini-swine model.Methods Twenty-three mini-swine with myocardial reperfusion injury were used as designed in the study protocol.About(3.54±0.90)×108 bone marrow mononuclear cells(MNC group,n=9) or(1.16± 1.07)×107 endothelial progenitor cells(EPC group,n=7) was infused into the affected coronary segment of the swine.The other mini-swine were infused with phosphate buffered saline as control(n=7).Echocardio-graphy and hemodynamic studies were performed before and 4 weeks after cell infusion.Myocardium infarction size was calculated.Stem cell differentiation was analyzed under a transmission electromicroscope.Results Left ventricular ejection fraction dropped by 0% in EPC group,2% in MNC group,and 10% in the control group 4 weeks after cell infusion,respectively(P<0.05).The systolic parameters increased in MNC and EPC groups but decreased in the control group.However,the diastolic parameters demonstrated no significant change in the three groups(P>0.05).EPC decreased total infarction size more than MNC did(1.60±0.26 cm2 vs.3.71±1.38 cm2,P<0.05).Undermature endothelial cells and myocytes were found under transmission electromicroscope.Conclusions Transplantation of either MNC or EPC may be beneficial to cardiac systolic function,but might not has obvious effect on diastolic function.Intracoronary infusion of EPC might be better than MNC in controlling infarction size.Both MNC and EPC may stimulate angiogenesis,inhibit fibrogenesis,and differentiate into myocardial cells.

IMPLANTATION OF AUTOLOGOUS BONE MARROW MONONUCLEAR CELLS INTO ISCHEMIC MYOCARDIUM ENHANCES CORONARY CAPILLARIES AND SYSTOLIC FUNCTION IN MINISWINE

Objective To investigate the therapeutic effectiveness of intracoronary implantation of autologous bone marrow mononuclear cells (BM-MNC) in miniswine model of reperfused myocardial infarction. Methods Sixteen miniswine myocardial ischemic reperfusion injury models made by ligation of the distal one third segment of left anterior descending artery for 90 minutes were randomized into 2 groups. In BM-MNC group (n = 9), (3.54±0.90)×108 BM-MNC were intracoronary injected, and in the control group (n = 7), phosphate buffered saline was injected by the same way. Echocardiographic and hemodynamic results, vessel density, and myocardial infarction size were evaluated and compared before and 4 weeks after cell transplantation. Results In BM-MNC group, there were no differences between before and 4 weeks after transplantation in aspects of left ventricular ejection fraction (LVEF), interventricular septal thickness, left ventricular lateral and anterior septal wall thickness, cardiac output, or +dp/dtmax. In control group, LVEF, interventricular septal thickness, left ventricular lateral and anterior septal wall thickness, cardiac output, and +dp/dtmax decreased significantly 4 weeks after transplantation (P < 0.05). Left ventricular end-diastolic pressure and –dp/dtmax did not change significantly before and after cell transplantation in both groups. Capillary density in BM-MNC group was greater than that in control group [(13.39 ± 6.96)/high power field vs. (3.50 ± 1.90)/high power field, P < 0.05]. Infarction area assessed by tetrazolium red staining and the infarction percentage decreased in BM-MNC group compared with those in control group (P < 0.05). Conclusions Transplantation of BM-MNC into myocardium with ischemic reperfusion injury increases capillary density and decreases infarction area. It has significantly beneficial effect on cardiac systolic function rather than on diastolic function.

TISSUE-ENGINEERED GRAFT CONSTRUCTED BY SELF-DERIVED BONE MARROW MONONUCLEAR CELLS AND HETEROGENEOUS ACELLULARIZED TISSUE MATRIX

Objective To create a method for constructing a tissue-engineered graft with self-derived bone marrow cells and heterogeneous acellular matrix.Methods The mononuclear cells were isolated from bone marrows drawn from piglets and cultured in different mediums including either vascular endothelial growth factor(VEGF)or platelet derived growth factor BB(PDGF-BB)to observe their expansion and differentiation.The aortas harvested from canines were processed by a multi-step decellularizing technique to erase.The bone marrow mononuclear cells cultured in the mediums without any growth factors were seeded to the acellular matrix.The cells-seeded grafts were incubated in vitro for 6 d and then implanted to the cells-donated piglets to substitute parts of their native pulmonary arteries.Results After 4 d culturing,the cells incubated in the medium including VEGF showed morphological feature of endothelial cells(ECs)and were positive to ECs-specific monoclonal antibodies of CD31,FLK-1,VE-Cadherin and vWF.The cells incubated in the medium including PDGF-BB showed morphological feature of smooth muscle cells(SMCs)and were positive to SMCs-specific monoclonal antibodies of α-SMA and Calponin.One hundred days after implantation of seeded grafts,the inner surfaces of explants were smooth without thrombosis,calcification and aneurysm.Under the microscopy,plenty of growing cells could be seen and elastic and collagen fibers were abundant.Conclusion Mesenchymal stem cells might exist in mononuclear cells isolated from bone marrow.They would differentiate into endothelial cells or smooth muscle cells in proper in vitro or in vivo environments.The bone marrow mononuclear cells might be a choice of seeding cells in constructing tissue-engineered graft.

Outcomes of autologous bone marrow mononuclear cell transplantation in decompensated liver cirrhosis

AIM:To determine the long-term efficacy of autologous bone marrow mononuclear cells(BM-MNCs)transplantation in terms of improving liver function and reducing complications in patients with decompensated cirrhosis.METHODS:A total of 47 inpatients with decompensated liver cirrhosis were enrolled in this trial,including32 patients undergoing a single BM-MNCs transplantation plus routine medical treatment,and 15 patients receiving medical treatment only as controls.Fortythree of 47 patients were infected with hepatitis B virus.Bone marrow of 80-100 mL was obtained from each patient and the BM-MNCs suspension was transfused into the liver via the hepatic artery.The efficacy of BM-MNCs transplantation was monitored during a24-mo follow-up period.RESULTS:Liver function parameters in the two groups were observed at 1 mo after BM-MNCs transfusion.Prealbumin level was 118.3±25.3 mg/L vs 101.4±28.7 mg/L(P=0.047);albumin level was 33.5±3.6g/L vs 30.3±2.2 g/L(P=0.002);total bilirubin 36.9±9.7 mmol/L vs 45.6±19.9 mmol/L(P=0.048);prothrombin time 14.4±2.3 s vs 15.9±2.8 s(P=0.046);prothrombin activity 84.3%±14.3%vs 74.4%±17.8%(P=0.046);fibrinogen 2.28±0.53 g/L vs1.89±0.44 g/L(P=0.017);and platelet count 74.5±15.7×109/L vs 63.3±15.7×109/L(P=0.027)in the treatment group and control group,respectively.Differences were statistically significant.The efficacy of BM-MNCs transplantation lasted 3-12 mo as compared with the control group.Serious complications such as hepatic encephalopathy and spontaneous bacterial peritonitis were also significantly reduced in BM-MNCs transfused patients compared with the controls.However,these improvements disappeared 24 mo after transplantation.CONCLUSION:BM-MNCs transplantation is safe and effective in patients with decompensated cirrhosis.It also decreases the incidence of serious complications.

Preventive effects of autologous bone marrow mononuclear cell implantation on intrahepatic ischemic-type biliary lesion in rabbits

BACKGROUND: The ischemic-type biliary lesion (ITBL) is one of the most serious biliary complications of liver transplantation. This study aimed to investigate the effects of autologous bone marrow mononuclear cell (BM-MNC) implantation on neovascularization and the prevention of intrahepatic ITBL in a rabbit model. METHODS: The rabbits were divided into control, experimental model, and cell implantation groups, with 10 in each group. The model of intrahepatic ITBL was established by clamping the hepatic artery and common bile duct. Autologous BM-MNCs were isolated from the tibial plateau by density gradient centrifugation and were implanted through the common hepatic artery. Changes in such biochemical markers as aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, gamma-glutamyltranspeptidase, total bilirubin and direct bilirubin were measured. Four weeks after operation, cholangiography, histopathological manifestations, differentiation of BM-MNCs, microvessel density and the expression of vascular endothelial growth factor were assessed. RESULTS: Compared with the experimental model group, the BM-MNC implantation group showed superiority in the time to recover normal biochemistry. The microvessel density and vascular endothelial growth factor expression of the implantation group were significantly higher than those of the control and experimental model groups. The ITBL in the experimental model group was more severe than that in the implantation group and fewer new capillary blood vessels occurred around it. CONCLUSIONS: Implanted autologous BM-MNCs can differentiate into vascular endothelial cells, promote neovascularization and improve the blood supply to the ischemic bile duct, and this provides a new way to diminish or prevent intrahepatic ITBL after liver transplantation. (Hepatobiliary Pancreat Dis Int 2010; 9:593-599)

Pre-degenerated peripheral nerves co-cultured with bone marrow-derived cells: a new technique for harvesting high-purity Schwann cells

Schwann cells play an important role in the peripheral nervous system, especially in nerve repair following injury, so artificial nerve regen- eration requires an effective technique for obtaining purified Schwann cells. In vivo and in vitro pre-degeneration of peripheral nerves have been shown to obtain high-purity Schwann cells. We believed that in vitro pre-degeneration was simple and controllable, and available for the clinic. Thus, we co-cultured the crushed sciatic nerves with bone marrow-derived cells in vitro. Results demonstrated that, 3 hours after injury, a large number of mononuclear cells moved to the crushed nerves and a large number of bone marrow-derived cells infiltrated the nerve segments. These changes promoted the degradation of the nerve segments, and the dedifferentiation and proliferation of Schwann cells. Neural cell adhesion molecule and glial fibrillary acidic protein expression were detected in the crushed nerves. Schwann cell yield was 9.08 ± 2.01 ×104/mg. The purity of primary cultured Schwann cells was 88.4 ± 5.79%. These indicate a successful new method for ob- taining Schwann cells of high purity and yield from adult crushed sciatic nerve using bone marrow-derived cells.

The effectiveness of the mononuclear fraction of autologous bone marrow in the treatment of experimental chronic limb ischemia

Objective To study of the effectiveness of using autologous mononuclear fraction of bone marrow for the treatment of chronic limb ischemia. Methods Results of autologous mononuclear fraction of bone marrow in 90 laboratory Wistar rats on a background of creating chronic limb ischemia was presented. Sampling was carried out from the bone marrow of the femur of the animal. The mononuclear fraction of bone marrow autologous 4 × 106 cells in a volume of 200 microliter were injected into the ischemic limb of the two points,in each of which 100 microliter:(1)Paravessel directly below the inguinal ligament at the level of the sacroiliac joint in the area of the anatomical location of collaterals in the projection of the internal iliac artery and its branches;(2)Intramuscularly in gastrocnemius muscle anterior-lateral surface of the middle third of the leg. Results In the experimental group of rats treated with autologous mononuclear fraction of bone marrow,the level of microcirculation compared with the intact group of animals on day 21 was higher than 6. 1% by day 28% ~ 31. 2%; compared with the control group-day 10 increased by 111% at day 21,85. 7% on day 28% ~ 97%. Conclusion Proposed method of treating pathogenically justified and can be recommended for use in clinical practice in the treatment of patients with chronic obliterating diseases of lower limb arteries.

三种途径移植自体BM-MNCs对猪急性心肌梗死后心功能影响的对比研究

目的对比研究心肌内、冠脉内及静脉内3种途径移植自体骨髓单个核细胞(BM-MNCs)对猪急性心肌梗死后心功能的影响。方法3种途径移植后4周观察小血管密度及心功能变化情况。结果(1)3种途径BM-MNCs移植后4周小血管密度均明显高于各自的对照组(P<0.01);心肌内移植BM-MNCs组与冠脉内移植BM-MNCs组相比小血管密度无统计学差异,但明显高于静脉内移植BM-MNCs组(P<0.01)。(2)3种途径BM-MNCs移植后4周LVEF及FS均明显高于各自对照组但组间比较无统计学差异;心肌内移植组和冠脉内移植组LVEDd均明显低于对照组,而静脉内移植组LVEDd虽低于对照组但无统计学差异(P>0.05)。结论3种途径移植BM-MNCs均有助于促进缺血心肌血管新生,改善左心室收缩功能,其中心肌内直接注射和冠脉内注射效果相似,静脉注射较前两种途径移植效果差;心肌内与冠脉内移植BM-MNCs组有减轻心室重构的作用,但在静脉移植组该作用不明显。

心肌、冠脉、静脉内移植BM-MNCs治疗猪急性心肌梗死的时机及安全性

目的:探讨心肌内、冠脉内及静脉内3种途径移植自体骨髓单个核细胞(BM-MNCs)治疗猪急性心肌梗死的时机及安全性。方法:分别于心肌梗死(MI)后即刻进行心肌内及静脉内移植,于MI后1周进行冠脉内移植。移植BM-MNCs后4周,观察小血管密度、心功能变化及冠脉侧支循环形成的情况,探讨移植的时机及安全性。结果:①3种途径BM-MNCs移植后4周,小血管的密度均明显高于各自的对照(P<0.01);心肌内移植与冠脉内移植相比,小血管的密度无统计学差异,但明显高于静脉内移植(P<0.01)。②3种途径移植BM-MNCs后4周,左室射血分数(LVEF)和缩短分数(FS)组相比较无统计学差异。心肌内移植和冠脉内移植的左室舒张末压(LVDEP)和左室舒张末期内径(LVEDD)均明显低于对照;而静脉内移植的LVDEP和LVEDD虽低于对照但无统计学差异。③3种途径移植BM-MNCs后1周,血清碱性成纤维细胞生长因子(bFGF)及血管内皮生长因子(VEGF)的浓度明显高于各自对照及术前水平(P<0.01)。④3种途径移植BM-MNCs后,均没有发现异常增生、钙化或肿瘤形成。结论:①心肌内及静脉内移植的时机应选在MI后即刻,冠脉内移植的时机应选在MI后1周。3种途径移植BM-MNCs后,均有助于促进缺血心肌中的血管新生,改善左室收缩功能。心肌内与冠脉内移植BM-MNCs均有改善左室舒张功能、减轻心室重构的作用,但在静脉内移植后该作用不明显。②3种途径移植BM-MNCs后,均没有发现明显的副作用。

Bone marrow-derived monocyte infusion improves hepatic fibrosis by decreasing osteopontin,TGF-β1,IL-13 and oxidative stress

AIM To evaluate the therapeutic effects of bone marrowderived CD11b+CD14+ monocytes in a murine model of chronic liver damage. METHODS Chronic liver damage was induced in C57BL/6 mice by administration of carbon tetrachloride and ethanol for 6 mo. Bone marrow-derived monocytes isolated by immunomagnetic separation were used for therapy. The cell transplantation effects were evaluated by morphometry,biochemical assessment,immunohistochemistry and enzyme-linked immunosorbent assay. RESULTS CD11b+CD14+ monocyte therapy significantly reduced liver fibrosis and increased hepatic glutathione levels. Levels of pro-inflammatory cytokines,including tumor necrosis factor-α,interleukin(IL)-6 and IL-1β,in addition to pro-fibrotic factors,such as IL-13,transforming growth factor-β1 and tissue inhibitor of metalloproteinase-1 also decreased,while IL-10 and matrix metalloproteinase-9 increased in the monocytetreated group. CD11b+CD14+ monocyte transplantation caused significant changes in the hepatic expression of α-smooth muscle actin and osteopontin. CONCLUSION Monocyte therapy is capable of bringing about improvement of liver fibrosis by reducing oxidative stress and inflammation,as well as increasing antifibrogenic factors.

Transplantation of autologous bone marrow mononuclear cells for patients with lower limb ischemia

Background Many treatment options for lower limb ischemia are difficult to apply for the patients with poor arterial outflow or with poor general conditions. The effect of medical treatment alone is far from ideal, especially in patients with diabetic foot. A high level amputation is inevitable in these patients. This study aimed to explore the effect of transplantation of autologous bone marrow mononuclear cells on the treatment of lower limb ischemia and to compare the effect of intra-arterial transplantation with that of intra-muscular transplantation. Methods In this clinical trial, 32 patients with lower limb ischemia were divided into two groups. Group 1 （16 patients with 18 affected limbs） received transplantation of autologous bone marrow mononuclear cells by intra-muscular injection into the affected limbs; and group 2 （16 patients with 17 affected limbs） received transplantation of autologous bone marrow mononuclear cells by intra-arterial injection into the affected limbs. Rest pain, coldness, ankle/brachial index （ABI）, claudication, transcutaneous oxygen pressure （tcPO2） and angiography （15 limbs of 14 patients） were evaluated before and after the mononuclear cell transplantation to determine the effect of the treatment. Results Two patients died from heart failure. The improvement of rest pain was seen in 76.5% （13/17） of group 1 and 93.3% （14/15） of group 2. The improvement of coldness was 100% in both groups. The increase of ABI was 44.4% （8/18） in group 1 and 41.2% （7/17） in group 2. The value of tcPO2 increased to 20 mmHg or more in 20 limbs. Nine of 15 limbs which underwent angiography showed rich collaterals. Limb salvage rate was 83.3% （15/18） in group 1 and 94.1% （16/17） in group 2. There was no statistically significant difference in the effectiveness of the treatment between the two groups. Conclusions Transplantation of autologous bone marrow mononuclear cells is a simple, safe and effective method for the treatment of lower limb ischemia, and the two approaches for the implantation, intra-muscular injection and intra-arterial injection, show similar results.

Effect of intracoronary autologous bone marrow mononuclear cells transplantation on arrhythmia in canines

Objective To observe the survival and the differentiation of grafted bone marrow cells(BM-MNCs)in host myocardium.To observe whether BM-MNCs transplantation can potentially cause arrhythmia and whether the BM-MNCs transplantation can alter the spatial distribution of connexins,important mediator for arrhythmia gen-

Mechanisms of improvement of left ventricle remodeling by transplanting two kinds of autologous bone marrow stem cells in pigs

Background The necrosis of a large number of myocardial cells after acute myocardial infarction （AMI） results in a decrease of cardiac function and ventricle remodeling. Stem cell transplantation could improve cardiac function after AMI, but the involving mechanisms have not been completely understood. The present study aimed to investigate the effects of transplantation of autologous bone marrow mononuclear cells （BM-MNC） and mesenchymal stem cells （MSCs） via the coronary artery on the ventricle remodeling after AMI as well as the mechanisms of the effects of transplantation of different stem cells on ventricle remodeling. Methods A total of 36 male pigs were enrolled in this study, which were divided into 4 groups： control group, simple infarct model group, BM-MNC transplantation group, and MSCs transplantation group. At 90 minutes when a miniature porcine model with AMI was established, transplantation of autologous BM-MNC （（4.7±1.7）×10^7） and MSCs （（6.2±1.6）×10^5） was performed in the coronary artery via a catheter. Ultrasound, electron microscope, immunohistochemical examination and real time reverse transcriptase-pelymerase chain reaction were used respectively to observe cardiac functions, counts of blood vessels of cardiac muscle, cardiac muscle nuclear factor （NF）-κB, myocardial cell apoptosis, and the expression of the mRNA of vascular endothelial growth factor （VEGF） and basic fibroblast growth factor （bFGF） in cardiac muscles. Multivariate Logistic regression was used to analyze the correlation factors of left ventricular end-diastolic diameter （EDD）. Results The number of blood vessels in the infarct zone and around its border in the BM-MNC transplantation group was more than those in the infarct model group and MSCs group （P=0.0001） and there was less myocardial cell apoptosis in the stem cell transplantation group than that in the infarct model group （all P 〈0.01）. The positive rate of NF-κB in the stem cell transplantation group was lower than that in the infarct model group （P=0.001）. The gene expression of VEGF in the infarct border zone of the BM-MNC group was higher than that in the MSCs group （P=0.0001）. The gene expression of bFGF in the infarct border zone in the MSCs transplantation group was higher than that in the infarct model group and the BM-MNC group （P=-0.0001）. Left ventricular ejection fraction was inversely proportional to the apoptotic rate of myocardial cells and cardiac muscle NF-κB but positively correlated with the number of blood vessels and the expression of VEGF and bFGF in the infarct zone and infarct border zone. The Multivariate Logistic regression analysis on the factors influencing the left ventricular end-diastolic diameter after stem cell transplantation showed that the expression of VEGF mRNA in the cardiac muscles in the infarct zone, the number of apoptotic myocardial cells and the expression of NF-κB in the infarct border zone were independent factors for predicting the inhibitory effect on the dilation of left ventricular EDD after stem cell transplantation. Conclusions Transplantation of autologous BM-MNC and MSCs in pigs can improve the condition of left ventricular remodeling and recover the cardiac functions after AMI. The improvement of cardiac functions is related to the increase of blood vessels, the increased expression of VEGF and bFGF, the reduction of myocardial cell apoptosis, and the decrease of NF-κB level in cardiac muscle tissues after stem cell transplantation.

Efficacy of autologous bone marrow mononuclear cell transplantation therapy in patients with refractory diabetic peripheral neuropathy

Background:Owing to the multifactorial nature of the pathogenesis of diabetic peripheral neuropathy (DPN),conventional drug therapies have not been effective.The application of stem cells transplantation may be useful for the treatment of DPN.This study was designed to assess the safety and therapeutic effects of autologous bone marrow mononuclear cells (BMMNCs) transplantation on the treatment of refractory DPN.Methods:One hundred and sixty-eight patients with refractory DPN were recruited and enrolled in the study.They received intramuscular injection of BMMNCs and followed at 1,3,6,12,18,24,and 36 months after the transplantation.Clinical data,Toronto Clinical Scoring System (TCSS),and nerve conduction studies (NCSs) were compared before and after the transplantation.Results:The signs and symptoms of neuropathy were significantly improved after BMMNCs transplantation.The values of the TCSS scores at 1 month (9.68 ± 2.49 vs.12.55 ± 2.19,P<0.001) and 3 months (8.47 ± 2.39 vs.12.55 ± 2.19,P<0.001) after the treatment reduced significantly compared with the baseline value.This decrement remained persistent until the end of the study.The conduction velocity and action potential and sensory nerves were significantly improved after transplantation (3 and 12 months after the treatment vs.the baseline:motor nerve conduction velocity,40.24 ± 2.80 and 41.00 ± 2.22 m/s vs.38.21 ± 2.28 m/s,P<0.001;sensory nerve conduction velocity,36.96 ± 2.26 and 39.15 ± 2.61 m/s vs.40.41 ± 2.22 m/s,P<0.001;compound muscle action potential,4.67 ± 1.05 and 5.50 ± 1.20 μV vs.5.68 ± 1.08 μV,P<0.001;sensory nerve action potential,4.29 ± 0.99 and 5.14 ± 1.26 μV vs.5.41 ± 1.14 μV,P<0.001).No adverse event associated with the treatment was observed during the follow-up period.Conclusions:Autologous transplantation of BMMNCs may be an effective and promising therapeutic strategy for the treatment of refractory DPN.

AMI患者使用经冠状动脉自体BM-MNC移植的长期效果随访观察

目的 探讨急性心肌梗死患者（AMI）采用经冠状动脉自体骨髓单个核细胞（BM-MNCs）移植治疗的长期疗效。方法 52例首次ST段抬高性AMI患者,在成功完成PCI术后,采用随机数字表法分为研究组和对照组各26例。研究组采用经冠状动脉注入BM-MNCs治疗,对照组同样方法注入等量生理盐水,比较两组患者治疗前与治疗后不同时间的随访观察结果差异。结果 术前研究组和对照组的左室射血分数（LVEF%）、左室舒张末期容积（LVEDV）、心肌缺血面积、心排指数（CI）、每博量指数（SI）、二尖瓣口舒张早期峰值流速（E）/二尖瓣口舒张晚期峰值流速（A）（E/A）测定值差异无统计学意义（P〉0.05）;术后6个月、24个月研究组的LVEF%、LVEDV、心肌缺血面积、CI、SI、E/A测定值均显著优于对照组（P〈0.05）;手术后第6个月、24个月研究组和对照组的LVEF%、LVEDV、心肌缺血面积、CI、SI、E/A测定值较术前均显著好转（P〈0.05）。研究组随访时间为16~24个月,平均随访时间为21.3±2.6个月,对照组随访时间15~24个月,平均随访时间20.9±3.0个月,两组患者的随访时间差异无具有统计学意义（P〉0.05）。随访过程中,研究组心血管事件发生率为11.54%（3/26）显著低于对照组的34.62%（9/26）（P〈0.05）。在随访过程中研究组有1例患者死亡,对照组3例患者死亡,两组患者的病死率比较差异无统计学意义（P〉0.05）。结论 采用经冠状动脉自体骨髓单个核细胞移植治疗对于保护AMI患者的心功能、降低患术后心血管事件的发生率具有积极意义。

Autologous bone marrow mononuclear cell transplantation in patients with chronic traumatic brain injury-a clinical study

Background:Chronic Traumatic Brain Injury(TBI)is one of the common causes of longterm disability worldwide.Cell transplantation has gained attention as a prospective therapeutic option for neurotraumatic disorders like TBI.The postulated mechanism of cell transplantation which includes angiogenesis,axonal regeneration,neurogenesis and synaptic remodeling,may tackle the pathology of chronic TBI and improve overall functioning.Methods:To study the effects of cell transplantation,50 patients with chronic TBI were enrolled in an open label nonrandomized study.The intervention included intrathecal transplantation of autologous bone marrow mononuclear cells and neurorehabilitation.Mean follow up duration was 22 months.Fifteen patients underwent second dose of cell transplantation,6 months after their first intervention.Percentage analysis was performed to analyze the symptomatic improvements in the patients.Functional independence measure(FIM)was used as an outcome measure to evaluate the functional changes in the patients.Statistical tests were applied on the pre-intervention and post-intervention scores for determining the significance.Comparative Positron Emission Tomography-computed tomography(PET CT)scans were performed in 10 patients to monitor the effect of intervention on brain function.Factors such as age,multiple doses,time since injury and severity of injury were also analyzed to determine their effect on the outcome of cell transplantation.Adverse events were monitored throughout the follow up period.Results:Overall 92%patients showed improvements in symptoms such as sitting and standing balance,voluntary control,memory,oromotor skills lower limb activities,ambulation,trunk&upper limb activity,speech,posture,communication,psychological status,cognition,attention and concentration,muscle tone,coordination,activities of daily living.A statistically significant(at p≤0.05 with p-value 0)improvement was observed in the scores of FIM after intervention on the Wilcoxon signed rank test.Better outcome of the intervention was found in patients with mild TBI,age less than 18 years and time since injury less than 5 years.Ten patients who underwent a repeat PET CT scan brain showed improved brain metabolism in areas which correlated to the symptomatic changes.Two patients had an episode of seizures which was managed with medication.They both had an abnormal EEG before the intervention and 1 of them had previous history and was on antiepileptics.No other major adverse events were recorded.Conclusion:This study demonstrates the safety and efficacy of cell transplantation in chronic TBI on long term follow up.Early intervention in younger age group of patients with mild TBI showed the best outcome in this study.In combination with neurorehabilitation,cell transplantation can enhance functional recovery and improve quality of life of patients with chronic TBI.PET CT scan brain should be explored as a monitoring tool to study the efficacy of intervention.