Uremia causes endothelial progenitor cell deficiency

BACKGROUND: Circulating bone marrow-derived endothelial progenitor cells (EPCs) promote vascular repair. Their number in peripheral blood correlates with endothelial function and cardiovascular risk in humans. We explored whether uremia influences the number of EPCs. METHODS: We assessed circulating CD34+ hematopoietic progenitor cells in whole blood using flow cytometry and EPCs (in vitro assay) in 46 patients with advanced renal failure and in 46 age- and gender-matched healthy subjects. Further, the effect of uremia on EPC differentiation was studied in vitro and in vivo. RESULTS: Both in renal patients (r= 0.34, P < 0.02) and in healthy subjects (r= 0.32, P= 0.04) the number of EPCs was significantly correlated to the absolute number of CD34+ hematopoietic progenitor cells. Renal patients had significantly fewer EPCs than healthy subjects, however (167 +/- 15 cells/high power field vs. 235 +/- 17 cells/high power field; P < 0.05). Uremic serum significantly (P < 0.05) inhibited EPC differentiation and functional activity in vitro. Amelioration of uremia after institution of renal replacement therapy in patients with terminal renal failure also significantly (P < 0.05) increased the number of EPCs. CONCLUSION: Uremia inhibits differentiation of EPCs. This may impair cardiovascular repair mechanisms in patients with renal failure.     

人脐血中血管内皮祖细胞体外扩增和鉴定的实验研究

目的：探讨从人脐血中分离、培养、体外扩增血管内皮祖细胞(EPC)的方法及体外进行EPC的鉴定。方法：采用密度梯度离心法从脐血中分离EPC，体外分别培养在包被有FN和不包被FN的培养皿中，采用免疫组织化学技术(SABC法)鉴定培养贴壁细胞表面标志CD31、CD34及KDR的表达；流式细胞仪检测细胞表面标志CD133及表达CD133细胞比例。结果：包被FN的培养皿中细胞贴壁及增殖均比未包被的多，免疫组化染色CD31、CD34及KDR均呈阳性，培养第4天流式细胞仪检测CD133^ 占21．3％。结论：密度梯度离心法可用于体外分离脐血中EPC进行实验研究，FN对于体外培养EPC有非常重要的作用。     

Podocyte detachment and reduced glomerular capillary endothelial fenestration in human type 1 diabetic nephropathy

The aim of this study was to investigate the structural characteristics of podocytes and endothelial cells in diabetic nephropathy. We studied 18 patients with type 1 diabetes (seven normoalbuminuric, six microalbuminuric, and five proteinuric), and six normal control subjects. Groups were not different for age. Type 1 diabetic groups were not different for diabetes duration or age at diabetes onset. Podocyte foot process width (FPW), fraction of glomerular basement membrane (GBM) surface with intact nondetached foot processes (IFP), fraction of glomerular capillary luminal surface covered by fenestrated endothelium [S(S)(Fenestrated/cap)] and classic diabetic glomerulopathy lesions were morphometrically measured. Albumin excretion (AER) and glomerular filtration (GFR) rates were also measured. GFR correlated inversely and AER directly with GBM and mesangial measurements in diabetic patients. FPW correlated inversely with GFR (r = -0.71, P = 0.001) and directly with AER (r = 0.66, P = 0.003), GBM, and mesangial parameters. The GBM fraction covered by IFP was decreased in proteinuric versus control subjects (P = 0.001), normoalbuminuric patients (P = 0.0002) and microalbuminuric patients (P = 0.04) and correlated with renal structural and functional parameters, including AER (r = -0.52, P = 0.03). Only 78% of GBM was covered by IFP in proteinuric patients. S(S)(Fenestrated/cap) was reduced in normoalbuminuric (P = 0.03), microalbuminuric (P = 0.03), and proteinuric (P = 0.002) patients versus control subjects. S(S)(Fenestrated/cap) correlated with mesangial fractional volume per glomerulus (r = -0.57, P = 0.01), IFP (r = 0.61, P = 0.007), and FPW (r = -0.58, P = 0.01). These novel studies document that podocyte detachment and diminished endothelial cell fenestration are related to classical diabetic nephropathy lesions and renal function in type 1 diabetic patients and support a need for further studies of podocyte/GBM adherence and podocyte/endothelial cell functional interactions in diabetic nephropathy.     

Kidney graft function determines endothelial progenitor cell number in renal transplant recipients

BACKGROUND: Circulating bone marrow-derived endothelial progenitor cells (EPCs) promote vascular repair, and their number correlates with endothelial function and cardiovascular risk in humans. In uremic patients, the number of functionally active EPCs is reduced. Thus, we assessed EPCs in stable patients 6 months or more after renal transplantation. METHODS: We analyzed circulating CD34+ hematopoietic progenitor cells (HPCs) using flow cytometry and EPCs (in vitro assay) in 74 renal transplant patients (51.6+/-11.5 years; 46 males), 74 age-matched healthy subjects, and 29 patients with preterminal renal failure. RESULTS: EPC numbers were similar in renal transplant recipients and controls (232+/-92 vs. 250+/-103/high power field; n.s.), but were significantly higher than in uremic patients (160+/-97/high power field; P=0.004). In addition, transplant recipients had more HPCs than controls (2.71+/-1.65 vs. 1.99+/-1.12 /microl; P=0.004) and uremic patients (1.64+/-0.96/microl; P=0.001). EPCs in renal transplant recipients correlated significantly with graft function(that is, Cockcroft-Gault clearance [r=0.294; P=0.012]), but not with age or HPCs. Moreover, in the multiple regression analysis, graft function (r=0.332; P=0.01) and diastolic blood pressure (r=-0.278; P=0.03) were independent predictors of EPCs. In vitro, sera from renal transplant recipients with poor graft function significantly inhibited EPC differentiation compared with sera from patients with a clearance above 50 mL/min (151+/-54 vs. 274+/-94 EPCs/high power field; P=0.02). CONCLUSIONS: EPC numbers in stable renal transplant recipients are comparable to those found in healthy subjects. In addition, graft function is a significant determinant of EPCs. Prospective studies should explore whether improvement of EPCs influences cardiovascular risk in renal transplant recipients.     

Cotreatment with darbepoetin and granulocyte colony-stimulating factor is efficient to recruit proangiogenic cell populations in patients with acute myocardial infarction

To determine whether newer combination cytokine treatment with granulocyte colony-stimulating factor (G-CSF) and darbepoetin can improve efficacy of stem cell therapy, we evaluated safety and peripheral blood stem/progenitor cell (PBSC) mobilizing effects of combination cytokine in comparison with G-CSF alone in patients with acute myocardial infarction (AMI). We randomized 60 patients with AMI into two groups under 2:1 ratio; combination treatment with darbepoetin and G-CSF (n = 41: Combicytokine group) and the G-CSF alone (n = 19: G-CSF group). After coronary angioplasty, G-CSF was treated for 3 days with dose of 10 μg/kg/day in both groups. Only in the combicytokine group, additional single intravenous injection of 4.5 μg/kg of darbepoetin was administrated immediate after coronary angioplasty. Combination cytokine treatment was well tolerated as was G-CSF alone. PBSCs were obtained by apheresis for intracoronary infusion after completion of cytokine treatment and were analyzed by flow cytometry. The purity of proangiogenic cells was higher in combination cytokine group than the G-CSF group. Specifically, proportion of CD34(+)/KDR(+) endothelial progenitor cells, CD3(+)/CD31(+) angiogenic T cells and Tie2(+)/CXCR4(+) cells in apheresis products were higher in the combicytokine group. These meant that the combicytokine treatment recruited PBSCs in higher purity and fewer unwanted inflammatory cells than G-CSF alone in apheresis products. Combination treatment with darbepoetin and G-CSF is safe and more efficient to mobilize and recruit proangiogenic cells than G-CSF alone in patients with AMI. (Trial registration: www.ClinicalTrials. gov identifier: NCT00501917).     

An assessment of P-glycoprotein expression and activity in peripheral blood lymphocytes of transplant candidates

BACKGROUND: P-glycoprotein (P-gp) is involved in the transport of the xenobiotic immunosuppressive agents and many cytokines, such as IL-2 and IFN-gamma. Hence, P-gp activity on peripheral blood lymphocytes (PBLs) could affect the pharmacologic response to xenobiotic immunosuppressants and immune responsiveness. The objectives of this study were to (1) determine the level of P-gp expression and activity on PBLs of kidney transplant candidates; and (2) determine whether P-gp expression correlates with P-gp activity. METHODS: We measured P-gp expression and activity on CD3(+)/CD8(+), CD3(+)/CD4(+), B lymphocytes, and NK cells of 36 kidney transplant candidates using a flow cytometric assay. P-gp activity was determined for each subpopulation of cells by the ratio of the mean Rhodamine 123 fluorescence (MFI Rh123) in the presence of verapamil divided by the MFI Rh123 in the absence of verapamil. P-gp expression was noted as the percentage of P-gp(+) cells. RESULTS: NK cells exhibited the greatest amount of P-gp activity (MFI Rh123 = 20.2 +/- 16.4) compared with other cell populations (P < .05). P-gp efflux activity was also significantly elevated in CD3(+)/CD8(+) cells (13.9 +/- 10.5) compared with B lymphocytes (4.9 +/- 2.7; P < .05) and CD4/CD3(+) cells (2.4 +/- 1.0; P < .05). P-gp expression was significantly higher in B lymphocytes (11.7 +/- 9.5) and NK cells (10.2 +/- 7.3) when compared with CD3(+)/CD8(+) cells (7.3 +/- 6.9) and CD3(+)/CD4(+) cells (6.4 +/- 3.8). P-gp expression was highly variable and did not correlate with P-gp activity (P > .05). CONCLUSIONS: CD3(+)/CD8(+) cells and NK cells, exhibited significantly increased P-gp activity compared with the other cell populations. P-gp expression is not a good correlate of P-gp activity. These findings may have important implications for the use of immunosuppressive drugs posttransplant and immune responsiveness after transplantation.     

腹腔镜结直肠癌根治对机体免疫状态的影响

目的探讨腹腔镜手术治疗结直肠癌对机体免疫状态的影响。方法2004年3月至2004年12月,同一手术组将可手术治疗的结直肠癌患者60例,随机分为腹腔镜手术组和开放手术组,每组各30例。分别在术前1d和术后3、7d取外周静脉血,测定C反应蛋白(CRP)、免疫球蛋白(Ig)A、IgM、IgG,CD3+、CD4+、CD8+、自然杀伤细胞(NK细胞)和CD4+CD45RA+、CD4+CD45RO+细胞及血淋巴细胞数并进行比较。结果开放手术组术后3d外周血淋巴细胞数(1.09±0.29)×109/L,CD4+细胞(0.54±0.14)×109/L,CD8+细胞(0.31±0.08)×109/L,CD4+CD45RO+细胞(61.1±8.9)%,IgM(136.9±52.8)IU/ml,IgG(115.2±45.7)IU/ml;术后7dCD8+细胞(0.32±0.09)×109/L,CD4+CD45RO+细胞(63.2±9.1)%,均明显低于术前(P<0.05,P<0.01)。而腹腔镜手术组术后3d除CD4+CD45RO+细胞犤(62.7±12.5)%犦较术前降低(P<0.05)外,其余免疫学指标无明显降低;而淋巴细胞数犤(1.29±0.37)×109/L犦、IgM犤(164.5±48.2)IU/ml犦和CD8+细胞数犤(0.38±0.09)×109/L犦与开放手术组比较,差异有统计学意义(P<0.05)。结论腹腔镜手术治疗结直肠癌比开放手术在免疫功能保护上更具有优势。     

Endothelial microparticles correlate with erectile dysfunction in diabetic men

Cell-derived microparticles are supposed to be involved in endothelial dysfunction and atherogenesis. This study aimed to evaluate circulating microparticles in diabetic subjects with erectile dysfunction (ED) and their relation with endothelial dysfunction. Thirty diabetic men with ED and 20 age-matched control subjects without ED were assessed for circulating microparticles and endothelial dysfunction. Flow cytometry was used to assess microparticles by quantification of circulating endothelial (EMP, CD31(+)/CD42b(-)) and platelet (PMP, CD31(+)/CD42b(+)) microparticles in peripheral blood. Endothelium-dependent flow-mediated dilation (FMD) was evaluated in the right brachial artery after reactive hyperemia. Compared with non-diabetic subjects, diabetic men presented significantly higher numbers of EMP (P=0.001), and reduced FMD (P=0.01), with a significant inverse correlation between the number of circulating EMP and the International Index of Erectile Function (IIEF) score (r=-0.457, P=0.01). Multivariate analysis correcting for age, anthropometric indices, glucose and lipid parameters, FMD and PMP identified EMP as the only independent predictor for IIEF score (P=0.03). EMP are elevated in impotent diabetic subjects and independently involved in the pathogenesis of ED.     

The extent of thermal injury affects fractions of mononuclear cells

Background

The mononuclear cell (MNC) fraction contains a variety of cell types, including stem cells such as endothelial progenitor cells (EPCs). EPC can rapidly revascularise ischaemic areas, but their role in burns is unclear.

Aim

This study investigates how thermal injury to the skin might influence mononuclear cells, CD34⁺ cells and circulating EPC.

Methods

The study group comprised 17 people with burns and 17 age-matched controls. Blood samples were collected at five different time points during the first 5 days of hospitalisation. Clinical parameters and scores were documented as well as cell counts for MNC, CD34⁺ cells and EPC. Counts were quantified by fluorescence-activated cell sorting. Serum was tested for vascular endothelial growth factor VEGF¹⁶⁵ by ELISA.

Results

All cell populations displayed significant, differing changes in counts and percentages after burn. These effects varied markedly over time and expressed different patterns if clinical scores were subjected to significance testing. EPC counts were significantly lowered in cases with fatal outcome.

Conclusion

Burn affects the numbers of circulating MNC, CD34⁺ and EPC. These time-dependent changes imply involvement of these cell groups in the trauma. EPC counts seem to be a predictive factor for outcome of cases of severe burn.     

Clinical and histologic determinants of renal outcome in ANCA-associated vasculitis: A prospective analysis of 100 patients with severe renal involvement

This study aimed to identify clinical and histologic prognostic indicators of renal outcome in patients with ANCA-associated vasculitis and severe renal involvement (serum creatinine >500 micromol/L). One hundred patients who were enrolled in an international, randomized, clinical trial to compare plasma exchange with intravenous methylprednisolone as an additional initial treatment were analyzed prospectively. Diagnostic renal biopsies were performed upon entry into the study. Thirty-nine histologic and nine clinical parameters were determined as candidate predictors of renal outcome. The end points were renal function at the time of diagnosis (GFR0) and 12 mo after diagnosis (GFR12), dialysis at entry and 12 mo after diagnosis, and death. Multivariate analyses were performed. Predictive of GFR0 were age (r = -0.40, P = 0.04), arteriosclerosis (r = -0.53, P = 0.01), segmental crescents (r = 0.35, P = 0.07), and eosinophilic infiltrate (r = -0.41, P = 0.04). Prognostic indicators for GFR12 were age (r = -0.32, P = 0.01), normal glomeruli (r = 0.24, P = 0.04), tubular atrophy (r = -0.28, P = 0.02), intraepithelial infiltrate (r = -0.26, P = 0.03), and GFR0 (r = 0.29, P = 0.01). Fibrous crescents (r = 0.22, P = 0.03) were predictive of dialysis at entry. Normal glomeruli (r = -0.30, P = 0.01) and treatment arm (r = -0.28, P = 0.02) were predictive of dialysis after 12 mo. No parameter predicted death. Both chronic and acute tubulointerstitial lesions predicted GFR12 in severe ANCA-associated glomerulonephritis, whereas plasma exchange was a positive predictor of dialysis independence after 12 mo for the entire patient group. Plasma exchange remained a positive predictor when patients who were dialysis dependent at presentation were analyzed separately (r = -0.36, P = 0.01). Normal glomeruli were a positive predictor of dialysis independence and improved renal function after 12 mo, indicating that the unaffected part of the kidney is vital in determining renal outcome.     

人脐血CD133+血管内皮祖细胞的培养、鉴定与分化研究

目的探讨从人脐血中分离、体外培养CD133^＋血管内皮祖细胞（EPCs）的方法及其生长特性和鉴定。方法采用密度梯度离心法结合MACS分选法提取人脐血中CD133^＋细胞,进行流式细胞仪检测纯度,予EBM-2培养液接种培养,观察其生长特性;利用Dil-LDL及FITC-Lectin摄取实验、细胞免疫组织化学等进行鉴定。结果经流式细胞仪检测CD133^＋细胞平均占单核细胞的（1．13±0．10）％,磁珠分选所得CD133^＋细胞平均纯度为（91．45±1．04）％;CD133^＋细胞贴壁生长,可分化为梭形血管内皮细胞及形成集落;CD133^＋细胞培养过程中Dil-LDL及FITC-Lectin摄取阳性,双染阳性率为（95．83±1．72）％;CD133^＋细胞培养1周后经免疫组织化学检测CD34、Ⅷ因子阳性率分别为（95．83±2．23）％和（95．92±1．43）％,与人脐静脉内皮细胞比较差异无统计学意义;CD133^＋细胞体外培养4d、1周可形成小血管结构。结论免疫磁珠分选法可获取较高纯度CD133^＋血管内皮祖细胞,在生长因子作用下诱导其分化为成熟血管内皮细胞。     

Markers of endothelial cell activation/injury: CD146 and thrombomodulin are related to adiponectin in kidney allograft recipients

BACKGROUND: Adiponectin may be used for assessing the risk of coronary artery disease (CAD) and may be related to the development of acute coronary syndrome. Decreased adiponectin has been associated with some risk factors for cardiovascular diseases such as male sex, obesity and diabetes mellitus. Adiponectin has antiatherogenic properties and attenuates endothelial inflammatory responses. CD146, a novel cell adhesion molecule, is localized at the endothelial junction. In kidney allograft recipients, endothelial dysfunction and atherosclerosis are almost universal. The aim of this cross-sectional study was to evaluate possible relations between adiponectin, CD146, and other markers of endothelial cell injury in 82 stable kidney transplant recipients (mean age 45 years, mean time after transplantation 47 months) with and without CAD. METHODS: Adiponectin and markers of endothelial injury: CD146, von Willebrand factor, thrombomodulin, ICAM, CD40L, P-selectin and other hemostatic markers were assessed using commercially available kits. RESULTS: Patients with CAD had evidence of more pronounced endothelial dysfunction, procoagulant state and lower adiponectin than patients without CAD. Adiponectin correlated significantly, in univariate analysis, with CD146 (r = 0.29, p = 0.009), thrombomodulin (r = 0.37, p = 0.001), protein Z (r = -0.25, p = 0.03), BMI (r = -0.26, p = 0.047), serum creatinine (r = 0.26, p = 0.02) and urea (r = 0.38, p = 0.001). CD146 correlated significantly with von Willebrand factor (r = 0.33, p = 0.002), thrombomodulin (r = 0.25, p = 0.025), age (r = 0.34, p = 0.001), platelets (r = -0.33, p = 0.002), serum urea (r = 0.24, p = 0.039), cholesterol (r = 0.24, p = 0.046), ICAM (r = 0.23, p = 0.036), protein C activity (r = -0.26, p = 0.019) and tended to correlate with serum creatinine and time after transplantation. In multivariate linear regression, independent predictors of adiponectin were CD146, thrombomodulin and urea, and of CD146 was mainly age of patients. CONCLUSIONS: Endothelial dysfunction and procoagulant state are more pronounced in kidney transplant recipients with CAD, particularly in those with lower GFR. In kidney transplant recipients, markers of endothelial cell injury are significantly increased relative to healthy volunteers. Elevation of adiponectin may be a defense mechanism against endothelial damage, reflected by elevated CD146 and thrombomodulin.     

VEGF gene transfer mobilizes endothelial progenitor cells in patients with inoperable coronary disease

BACKGROUND: Direct transfection of ischemic myocardium with naked plasmid DNA encoding for vascular endothelial growth factor-165 (VEGF165) has been shown to mobilize endothelial progenitor cells (EPCs). This study examined the kinetics of circulating EPCs isolated from peripheral blood mononuclear cells after gene transfer, and their role in neovascularization of ischemic myocardium. METHODS: The mononuclear cell population was isolated from peripheral venous blood samples of patients with functional class III or IV angina receiving intramyocardial VEGF165 gene transfer. Peripheral blood mononuclear cells were examined by an in vitro EPC culture assay and fluorescent-activated cell sorting. The data were compared with a control group consisting of patients who had undergone off-pump coronary artery bypass grafting without receiving gene transfer. RESULTS: Coinciding with a rise in VEGF levels, mobilization of EPCs increased significantly over base line for 9 weeks after the treatment (121+/-14 cells/mm2 versus 36.8+/-8 cells/mm2, p < 0.0005), followed by a subsequent decrease. Fluorescent-activated cell sorting analysis confirmed culture assay data, with a statistically significant rise in cells expressing vascular endothelial-cadherin, CD51/61 [alphavbeta3], CD62E [E-selectin], CD34, and KDR. The control group failed to show significant mobilization of EPCs. CONCLUSIONS: Mobilization of EPCs with resultant postnatal vasculogenesis, may play a role in revascularizing ischemic myocardium following human gene transfer with VEGF165.     

The mobilisation of mononuclear cells and endothelial progenitor cells after burn injury in a porcine model

Background

Mononuclear blood cells (MNCs) consist of heterogeneous cell populations, for example, CD34+ cells and endothelial progenitor cells (EPCs). EPCs are involved in vasculogenesis, but little is known about their role during burn trauma.

Aim

This study investigates the role of MNCs and their subpopulations during and after burn injury in an experimental porcine setting.

Methods

Eighteen 8-week-old German land pigs were scalded by immersion in 70 °C hot water for 3 min, resulting in a 30% total body surface area (TBSA) full-thickness burn. Vascular endothelial growth factor (VEGF) serum concentrations and MNC, EPC and CD34+ cell counts were measured at eight different time points up to 48 h following trauma.

Results

The experimental porcine setting made it possible to determine the cell counts of MNCs, EPCs and CD34+ cells directly during burn trauma, which has not been described before. The data revealed a fulminant drop in MNC and EPC during burn trauma, whereas the CD34+ cell fraction rose. Besides significant changes in the VEGF serum concentration, a correlation between VEGF and EPC was also observed.

Conclusion

The results show that MNCs and their subpopulations are significantly affected by burn trauma and underpin their potential diagnostic and therapeutic importance during and after burn injury.     

Factors influencing T-lymphopoiesis after allogeneic hematopoietic cell transplantation

In adult recipients of allogeneic hematopoietic cell transplants (HCT) studied at 1 year after grafting, there was a significant correlation between the counts of T cell receptor excision circle (TREC)-containing CD4 T cells (presumed recent thymic emigrants) and the counts of total T cells (r=0.65, P<0.001). Thus, the reconstitution of CD4 T cell pool depends on T cell generation from hematopoietic stem cells (T-lymphopoiesis). We evaluated factors that could affect T-lymphopoiesis. Low TREC-containing CD4 T cell counts were associated with older patient age (r=-0.41, P=0.01) but not with donor age, graft type (marrow vs. blood stem cells), CD34 cell dose, conditioning (with vs. without irradiation), acute graft-versus-host disease (aGVHD), or chronic graft-versus-host disease (cGVHD) in multivariate analysis. We conclude that patient age is the primary determinant of CD4 T-lymphopoiesis after allogeneic HCT.     

Transplantation of endothelial progenitor cells transferred by vascular endothelial growth factor gene for vascular regeneration of ischemic flaps

BACKGROUND: Neovascularization occurs through two mechanisms: angiogenesis and vasculogenesis. Therefore, there are two strategies to promote neovascularization: therapeutic angiogenesis and therapeutic vasculogenesis (endothelial progenitor cells therapy). MATERIALS AND METHODS: In this study, we examined whether or not endothelial progenitor cells combined with vascular endothelial growth factor (VEGF) gene therapy is useful for ischemia surgical flaps in vivo. At the same time, we quantitatively compared the neovascularization ability of transplanted endothelial progenitor cells (EPCs) transducted with VEGF165 gene and EPCs alone. EPCs were isolated from cord blood of healthy human volunteers, cultured in vitro for 7 days and identified by immunofluorescence. After transduced with VEGF165 gene in vitro, proliferative activity of EPCs was assessed using MTT assay. CM-DiI was used to trace EPCs in vivo 4 days after injection of 5 x 10(5) VEGF-transduced EPCs(VEGF-transduced EPCs group, n = 10), 5 x 10(5) EPCs (non-transduced EPCs group, n = 10) in 500 microL EBM-2 media, or 500 microL EBM-2 media (EBM-2 media group, n = 10) local, a cranially based flap was elevated on the back of nude mice. The percent flap survival, neovasculariztion and blood flow recovery of flaps was detected. RESULTS: EPCs expressed cell markers CD34, KDR, and CD133. A statistically significant increase in percent flap survival was observed in mice of VEGF-transduced EPCs group as compared with that of non-transduced EPCs group: 67.99 +/- 6.64% versus 59.43 +/- 4.69% (P < 0.01), and 41.24 +/- 2.44% in EBM-2 media group (P < 0.01). The capillary density and blood flow recovery of flaps in VEGF-transduced EPCs group were both improved. CM-DiI-labeled VEGF-transduced EPCs were observed in vivo and the numbers of cells increased. CONCLUSION: EPCs from human cord blood can increased neovascularization of ischemic flaps and augmented the survival areas, and VEGF-transduced EPCs have more powerful ability of promoting neovascularization in animal model of ischemic flaps.     

Arterial erectile dysfunction: reliability of penile Doppler evaluation integrated with serum concentrations of late endothelial progenitor cells and endothelial microparticles

We recently showed the diagnostic value of a new immunophenotype of blood endothelial progenitor cells (EPC) (CD45(-)/CD34(+)/CD144(+)) and endothelial microparticles (EMP) (CD45(-)/CD144(+)/annexin V(+)) in patients with arterial erectile dysfunction (AED), particularly in patients with associated late-onset hypogonadism and/or metabolic syndrome. In addition, we evaluated the effects of androgen replacement therapy, aerobic physical activity, and tadalafil administration on these markers. The aim of this study was to evaluate the serum concentrations of EPCs and EMPs in a large cohort of patients with AED according to severity of cavernous arterial insufficiency evaluated by penile Doppler. A total of 120 patients (aged 58.0 ± 6.0 years) with AED were enrolled in this study. Patients were classified into 3 groups based on value of peak systolic velocity (PSV). Group A: 37 patients with PSV <25 cm/s (severe arterial insufficiency); group B: 40 patients with PSV between 25 and 29 cm/s (moderate arterial insufficiency); group C: 43 patients with PSV between 30 and 34 cm/s (mild arterial insufficiency). Twenty patients (aged 60.0 ± 3.0 years) with psychogenic erectile dysfunction (PED) represented the control group. EPC and EMP blood concentrations were evaluated by flow cytometry. Patients with AED had significantly higher blood pressure, triglyceride levels, homeostasis model assessment index of insulin resistance, cavernous artery acceleration time, and intima-media thickness than those with PED, whereas International Index of Erectile Function score, high-density lipoprotein cholesterol level, and cavernous artery PSV were lower than those in PED. Both EPC and EMP levels were significantly higher in patients with AED compared with patients with PED. Among 3 groups of patients with AED, there were no significant differences in metabolic parameters examined, but group A showed significantly higher values of cavernous artery acceleration time and intima-media thickness than group B and group C. Finally, group A showed serum concentrations of EPCs and EMPs significantly higher compared with other groups with AED. Patients with AED showed worse metabolic parameters, cavernous artery parameters, and EPC and EMP levels compared with patients with PED. Among patients with AED, those with PSV <25 cm/s showed worse findings of endothelial dysfunction. This suggests that AED is an expression of endothelial damage and that this original immunophenotype of EPCs and EMPs may be considered a predictor of endothelial dysfunction in patients with AED. Finally, this study confirmed the reliability of penile Doppler evaluation integrated with these serum markers of endothelial dysfunction.     

紫绀型先天性心脏病病人循环内皮祖细胞的数量及功能变化

目的 探讨紫绀型先天性心脏病病人外周血内皮祖细胞(endothelial progenitor cell,EPC)的数量及功能特点.方法 选取法洛四联症及单纯室间隔缺损病人各10例,采用Ficoll密度梯度离心法从外周血中分离单个核细胞,用添加牛脑垂体提取物和胎牛血清的M199培基进行体外诱导分化培养.用免疫组化和透射电镜行EPC表型鉴定;采用流式细胞仪计数CD133+/KDR+细胞;采用改良的Boyden小室,黏附能力测定实验和MTT比色法,观察EPC的迁移、黏附及增殖能力.结果 紫绀型先天性心脏病病人外周血内皮祖细胞的数量增多[集落记数(14.7±3.1)/100倍视野对(8.2±1.3)/100倍视野;DiIAcldl+/UEA-Ⅰ+细胞数(72.2±9.73)/200倍视野对(51.2±3.83)/200倍视野;CD133+/KDR+细胞百分比(0.66±0.20)%对(0.18±0.08)%,P＜0.01],其迁移能力[(140.6±9.24)/200倍视野对(91.84±8.58)/200倍视野]、黏附能力[(149.00±11.58)/200倍视野对(112.6±7.02),200倍视野]及增殖能力(OD值0.34±0.02对0.27±0.01)增强(P＜0.01).结论 紫绀型先天性心脏病病人循环内皮祖细胞的数量增多,其迁移、黏附及增殖能力增强.     

Three monocyte-related determinants of atherosclerosis in haemodialysis.

BACKGROUND: It has been suggested that monocyte-related inflammatory mediators play a role in atherosclerosis. Haemodialysis induces phenotypic changes in adhesion molecule expression on monocytes. Soluble vascular cell adhesion molecule-1 (sVCAM-1), an adhesion molecule involved in monocyte recruitment, has been proposed to correlate with the extent of atherosclerosis in humans. Monocyte chemotactic protein-1 (MCP-1) functions as a monocyte-specific chemoattractant. METHODS: We studied monocyte count, CD11b/CD18 expression on monocytes, MCP-1, and sVCAM-1 in nine patients on either cuprophane or polysulphone haemodialysis (n=18 treatments) at times 0 (before haemodialysis), 3 h (end of haemodialysis), 4, 6, 8 and 24 h after start of treatment, as well as in 18 healthy subjects. RESULTS: Monocyte CD11b/CD18 expression increased with both membranes (P:<0.001) during and after dialysis compared to before treatment. The concentrations of sVCAM-1 and MCP-1 were higher in patients compared to those in controls both before, during and after haemodialysis (P:<0.001 at all time points). There were correlations between the expression of CD11b/CD18 on monocytes and the interdialytic concentrations of sVCAM-1 (r=0.76, P:<0.001) and MCP-1 (r=0.54, P:<0.05) and between MCP-1 and sVCAM-1 before and after haemodialysis (P:<0.05). CONCLUSION: Patients on haemodialysis have an increased systemic chemotactic activity for monocytes, unphysiological phenotypic alterations in CD11b/ CD18 expression during and after dialysis, and increased sVCAM-1 and MCP-1 concentrations. Prospective studies are needed to establish the role of these abnormalities in the pathogenesis of atherosclerosis in haemodialysis patients.     

Renal function and structure in albuminuric type 2 diabetic patients without retinopathy.

BACKGROUND: In type 2 diabetic patients without retinopathy the cause of albuminuria is heterogeneous and our knowledge of the relationship between kidney structure and function in these patients is limited. Therefore, a long-term study evaluating the structural-functional relationship in albuminuric type 2 diabetic patients without retinopathy was performed. METHODS: Mesangial volume of total glomerular volume (Vv (mes/glom)), fractional area of focal interstitial fibrosis and tubular atrophy of cortical area (FF) and percentage of sclerosed glomeruli (S/G) were measured on kidney biopsies from 49 type 2 diabetic patients without retinopathy. Glomerular filtration rate (GFR) was determined at least 3 times (median 8 (range 3-20)) in each patient. Patients were followed for 7.0 (1.1-17) years. Albuminuria and blood pressure were measured every 3-6 months. RESULTS: Biopsies revealed diabetic glomerulopathy (DG-group) in 69% of the patients (27 male/7 female) and normal glomerular structure (n=9) or glomerulonephritis (n=6) were found in 31% (13 male/2 female) (NDG-group). In the DG-group GFR decreased from 97+/-5 to 66+/-5 ml/min/1.73 m(2) (mean+/-SE) (P<0.001), with a rate of decline in GFR of 5.3+/-0.8 ml/min/year and in the NDG-group from 93+/-7 to 74+/-11 ml/min/1.73 m(2) (P<0.01), with a rate of decline in GFR of 3.2+/-0.9 ml/min/year, P=0.09 between groups. Mean arterial blood pressure decreased from 109+/-2 to 100+/-2 mm Hg (P<0.001) (DG-group) and remained unchanged in the NDG-group. An association between Vv (mes/glom) and rate of decline in GFR was revealed mainly in the NDG-group (DG-group; r=0.31, P=0.07 and NDG-group; r=0.74, P<0.01). Furthermore, the rate of decline in GFR seemed to be associated with FF in the NDG group (r=0.48, P=0.07). Percentage of S/G was not associated with the rate of decline in GFR. Vv (mes/glom) was associated with mean albuminuria during follow-up in the DG group; r=0.38, P<0.03 (NDG group; r=0.51, P=0.09). Albuminuria was an independent predictor of the rate of decline in GFR in both groups (DG-group; r=0.40, P<0.05 and NDG-group; r=0.61, P<0.01). CONCLUSIONS: Our study revealed a tendency to a faster rate of decline in GFR in the DG-group compared to the much smaller NDG-group, characterized by marked heterogeneity of the underlying kidney lesions and rate of GFR loss. A large mesangial volume fraction was associated with increased albuminuria and loss in GFR. Albuminuria acted as a progression promoter in both groups.