自体骨髓单个核细胞移植治疗大鼠后肢缺血

目的　探讨自体骨髓单个核细胞（BM-MNC）移植用于大鼠缺血后肢的治疗后实现血管再生的能力。方法　建立大鼠后肢缺血动物模型,将取于自体的BM-MNC制成细胞悬液注射于缺血部位,分别在移植后2和30d时行动脉造影,用免疫组化方法检测内皮祖细胞（EPC）,毛细血管密度以及测定血管内皮生长因子(VEGF)的表达。结果　缺血肌组织中的EPC含量增高（P<0.01）。BM-MNC移植组在移植早期VEGF表达显著增高（P<0.01）。细胞移植后30dBM-MNC移植组毛细血管密度明显高于其他组（P<0.01）,血管造影可见侧支循环建立。结论　自体骨髓单个核细胞移植于大鼠后肢缺血区能促进血管新生,改善侧支循环,可望成为一种简单有效的治疗下肢缺血的方法。     

Idiopathic dilated cardiomyopathy exhibits defective vascularization and vessel formation

BACKGROUND: Ultrastructural findings of idiopathic dilated cardiomyopathy (IDCM) include myocyte atrophy and myofilament loss, yet little is known about the vascular abnormalities present in IDCM. METHODS AND RESULTS: Patients with IDCM and controls underwent multi-slice CT to examine length and diameter of epicardial vasculature. The levels of mobilizing cytokines and circulating EPCs were assessed by endothelial colony formation assay and flow cytometry. Immunohistochemistry and Western blot were used to examine microvessel density and expression of HIF-1alpha and beta-catenin. Main epicardial coronary arteries were shorter and smaller, and microvascular density was reduced in the epicardium in IDCM. Epicardial vessel paucity was associated with increased numbers of HIF-1alpha(+) cells (46.8+/-13.1% vs. 19.4+/-9.4%, p=0.006) indicating local epicardial hypoxia and elevation of circulating VEGF-A (394 pg/mL vs. 22 pg/mL, p=0.001). The number of mobilized progenitors CD133(+)/VEGF-R2(+) was 21-fold higher in IDCM compared with controls (6.5+/-3.3% vs. 0.3+/-0.2%; p<0.001). Moreover, this defective vascularization was associated with reduced myocardial expression of vascular beta-catenin, an important angiogenic regulator. CONCLUSIONS: This study shows defective vascularization and impaired vasculogenesis (the de novo vascular organization of mobilized endothelial progenitors) and angiogenesis (by which new blood vessels are formed from pre-existing mature endothelial cells) in human IDCM.     

Impaired development and dysfunction of endothelial progenitor cells in type 2 diabetic mice

AimDysfunction of circulating endothelial progenitor cells (EPCs) has been shown to affect the development of microvascular diseases in diabetes patients. The aim of this study was to elucidate the development and mechanical dysfunction of EPCs in type 2 diabetes (T2D).MethodsThe colony-forming capacity of EPCs and differentiation potential of bone marrow (BM) c-Kit(+)/Sca-I(+) lineage-negative mononuclear cells (KSL) were examined in T2D mice, db/db mice and KKA^y mice, using EPC colony-forming assay (EPC-CFA).ResultsT2D mice had fewer BM stem/progenitor cells, and proliferation of KSL was lowest in the BM of db/db mice. In T2D mice, the frequency of large colony-forming units (CFUs) derived from BM-KSL was highly reduced, indicating dysfunction of differentiation into mature EPCs. Only a small number of BM-derived progenitors [CD34(+) KSL cells], which contribute to the supply of EPCs for postnatal neovascularization, was also found. Furthermore, in terms of their plasticity to transdifferentiate into various cell types, BM-KSL exhibited a greater potential to differentiate into granulocyte macrophages (GMs) than into other cell types.ConclusionT2D affected EPC colony formation and differentiation of stem cells to mature EPCs or haematopoietic cells. These data suggest opposing regulatory mechanisms for differentiation into mature EPCs and GMs in T2D mice.     

内皮祖细胞移植对糖尿病兔下肢缺血区血管生成相关基因表达的影响

目的研究内皮祖细胞（EPC）移植治疗糖尿病兔下肢缺血对血管生成相关基因表达的影响。方法日本大耳白兔16只，四氧嘧啶法建立糖尿病模型后，随机分为磷酸盐缓冲液（PBS）对照组与EPC治疗组，分别用PBS和骨髓来源经体外扩增的EPC移植于下肢缺血区，14天后用基因芯片分析EPC移植对血管生成相关基因表达的影响，同时检测肌浆中血管内皮生长因子（VEGF）浓度。结果EPC治疗组下肢缺血区27个血管生成基因表达上调超过2倍，移植治疗组VEGF的浓度显著高于对照组，差异有统计学意义。结论EPC植入糖尿病兔下肢缺血区，可以上调包括VEGF在内的多种血管生成相关基因，同时肌浆中VEGF水平升高。     

Significance and therapeutic potential of endothelial progenitor cell transplantation in a cirrhotic liver rat model

BACKGROUND & AIMS: We investigated whether endothelial progenitor cell (EPC) transplantation could reduce established liver fibrosis and promote hepatic regeneration by isolating rat EPCs from bone marrow cells. METHODS: Recipient rats were injected intraperitoneally with carbon tetrachloride (CCl(4)) twice weekly for 6 weeks before initial administration of EPCs. CCl(4) was then readministered twice weekly for 4 more weeks, and EPC transplantation was carried out for these same 4 weeks. RESULTS: At 7 days in culture, the cells expressed Thy-1, CD31, CD133, Flt-1, Flk-1, and Tie-2, suggesting an immature endothelial lineage. Immunohistochemical analyses showed fluorescent-labeled, transplantation EPCs were incorporated into the portal tracts and fibrous septa. Single and multiple EPC transplantation rats had reduced liver fibrosis, with decreased alpha2-(I)-procollagen, fibronectin, transforming growth factor-beta, and alpha-smooth muscle actin-positive cells. Film in situ zymographic analysis revealed strong gelatinolytic activity in the periportal area, in accordance with EPC location. Real-time polymerase chain reaction analysis of multiple EPC-transplantation livers showed significantly increased messenger RNA levels of matrix metalloproteinase (MMP)-2, -9 and -13, whereas tissue inhibitor of metalloproteinase-1 expression was significantly reduced. Expression of hepatocyte growth factor, transforming growth factor-alpha, epidermal growth factor, and vascular endothelial growth factor was increased in multiple EPC-transplantation livers, while hepatocyte proliferation increased. Transaminase, total bilirubin, total protein, and albumin levels were maintained in EPC-transplantation rats, significantly improving survival rates. CONCLUSIONS: We conclude that single or repeated EPC transplantation halts established liver fibrosis in rats by suppressing activated hepatic stellate cells, increasing matrix metalloproteinase activity, and regulating hepatocyte proliferation.     

An Experimental Study on the Dynamic Mechanical Properties of Epoxy Polymer Concrete under Ultraviolet Aging

Epoxy polymer concrete (EPC) is widely applied in engineering for its excellent mechanical properties. The impact loads and severe climatic conditions such as ultraviolet radiation, temperature change and rain erosion are in general for its engineering practice, potentially degrading the performance of EPC. In this paper, a procedure of accelerated aging for EPC, imitating the aging effect of ultraviolet radiation and hygrothermal conditions based on the meteorological statistics of Guangzhou city, was designed. After various periods of accelerated aging, the dynamic behaviors of EPC were studied by using a Split Hopkinson Pressure Bar (SHPB). The verification of the experimental data was performed. The two-stage dynamic compression stress-strain curves were obtained: (a) linear growth stage following by strain hardening stage at impact velocity 12.2 m/s and 18.8 m/s, (b) linear growth stage and then a horizontal stage when impact velocity is 25.0 m/s, (c) linear growth stage following by strain softening stage at impact velocity 29.2 m/s. The experimental results show that the specimens after longer accelerated aging tend to be more easily broken, especially at impact velocity 12.2 m/s and 18.8 m/s, while the strain rate is the main factor affecting the compression strength and stiffness. Ultimately the influence of strain rate and equivalent aging time on dynamic increase factor was revealed by a fitting surface.     

Circulating endothelial progenitor cells correlate with erectile function in patients with coronary heart disease.

AIMS: The aim of the study was to determine the influence of endothelial progenitor cells (EPC) on erectile dysfunction (ED). EPC play a major role in repair mechanisms of the endothelial monolayer, but the role of EPC in ED is unclear. METHODS AND RESULTS: Circulating levels of CD34(+)/KDR(+) and CD133(+) EPC were determined in 119 patients with known coronary artery disease. ED was evaluated with an ED-score generated from the KEED questionnaire. Prevalence of ED was 59.7%. In univariate analysis, age, hypertension, reduced left ventricular ejection fraction (LVEF), diabetes, and circulating levels of CD133(+) EPC, but not cardiovascular drug treatment were associated with ED. Body mass index (BMI) was positively (r = 0.319, P=0.003) and high-density lipoprotein was negatively (r=-0.246, P=0.034) correlated with ED. Adjustment for age, diabetes, hypertension, BMI, smoking, LVEF, use of statins and lower urinary tract symptoms, and prior coronary intervention revealed low levels of circulating immature CD133(+) EPC as independent risk factor for ED (95% CI -11.183 to -1.7371, P=0.008). CONCLUSION: Reduced levels of circulating CD133(+) EPC are an independent risk factor for ED. Thus, EPC may be a link between cardiovascular risk factors, endothelial dysfunction, and ED.     

乳腺包被性乳头状癌的临床病理分析

目的 探讨乳腺包被性乳头状癌（encapsulated papillary carcinoma, EPC）的临床病理学特征、诊断、治疗及预后。方法 回顾性分析新疆医科大学附属肿瘤医院病理科2010年1月-2013年12月间,诊断为囊内乳头状癌或包被性/包裹性乳头状癌35例及48例对照组导管内乳头状癌的临床病理资料及免疫表型。结果 EPC患者均为女性,年龄29~83岁,平均61岁,肿块大小平均2.4 cm (范围0.6~4 cm)。35例EPC中,22例为单纯性EPC、7例伴导管原位癌、4例伴微小浸润癌、2例伴非特殊类型浸润癌。35例EPC与48例导管内乳头状癌病变内部均未见肌上皮细胞,CK5/6及p63肌上皮染色结果显示,EPC病变周围肌上皮数量较导管内乳头状癌导管壁肌上皮数量明显减少,差异具有统计学意义（P<0.05）;35例EPC中80%激素受体阳性,5.71%HER2表达阳性。8例（22.86%）EPC患者行肿块切除,27例（77.14%）患者行乳房切除术,3例（8.57%）发生淋巴结转移。术后经随访2~48月,患者均存活。结论 乳腺包被性乳头状癌是一种好发于老年女性的恶性肿瘤,病变周缘肌上皮明显减少甚至缺如,单纯性EPC也可发生淋巴结转移,被认为是一种惰性的浸润癌,生物学行为介于原位癌与浸润癌之间。若单独发生或伴随原位癌及微小浸润癌时,应参照原位癌治疗, EPC伴随浸润癌时,应参照浸润癌的治疗标准进行。