Regulation of angiogenesis by a non-canonical Wnt-Flt1 pathway in myeloid cells

Myeloid cells are a feature of most tissues. Here we show that during development, retinal myeloid cells (RMCs) produce Wnt ligands to regulate blood vessel branching. In the mouse retina, where angiogenesis occurs postnatally, somatic deletion in RMCs of the Wnt ligand transporter Wntless results in increased angiogenesis in the deeper layers. We also show that mutation of Wnt5a and Wnt11 results in increased angiogenesis and that these ligands elicit RMC responses via a non-canonical Wnt pathway. Using cultured myeloid-like cells and RMC somatic deletion of Flt1, we show that an effector of Wnt-dependent suppression of angiogenesis by RMCs is Flt1, a naturally occurring inhibitor of vascular endothelial growth factor (VEGF). These findings indicate that resident myeloid cells can use a non-canonical, Wnt-Flt1 pathway to suppress angiogenic branching.     

A novel <Emphasis Type="Italic">in vitro</Emphasis> angiogenesis model based on a microfluidic device

Angiogenesis is very important for many physiological and pathological processes. However, the molecular mechanisms of angiogenesis are unclear. To elucidate the molecular mechanisms of angiogenesis and to develop treatments for "angiogenesis-dependent" diseases, it is essential to establish a suitable in vitro angiogenesis model. In this study, we created a novel in vitro angiogenesis model based on a microfluidic device. Our model provides an in vivo-like microenvironment for endothelial cells (ECs) cultures and monitors the response of ECs to changes in their microenvironment in real time. To evaluate the potential of this microfluidic device for researching angiogenesis, the effects of pro-angiogenic factors on ECs proliferation, migration and tube-like structure formation were investigated. Our results showed the proliferation rate of ECs in 3D matrix was significantly promoted by the pro-angiogenic factors (with an increase of 59.12%). With the stimulation of pro-angiogenic factors gradients, ECs directionally migrated into the Matrigel from low concentrations to high concentrations and consequently formed multi-cell chords and tube-like structures. These results suggest that the device can provide a suitable platform for elucidating the mechanisms of angiogenesis and for screening pro-angiogenic or anti-angiogenic drugs for "angiogenesis-dependent" diseases.     

三例眼部突变小鼠的培育及特征分析

摘要: 目的 筛选培育眼部突变表型小鼠,为人类相关疾病的研究提供材料。方法 采用 N-乙基-N-亚硝基脲 ( N-ethyl-N-nitrosourea,ENU) 诱变处理 G0 代小鼠,繁育获得 G1 代,筛选眼部突变表型个体,进行遗传力试验、临床 诊断及病理学观察。结果 本实验繁殖 G1 代小鼠 2782 只,经筛查获得眼部突变表型小鼠 65 只,可稳定遗传小鼠 3 例,分别表现为: 角膜混浊、小眼球和虹膜异常等特征。角膜混浊者其角膜症状严重程度差异较大,角膜病变部位 明显增厚,部分伴有新生血管; 小眼球者睑裂较小,甚至上下眼睑粘连,外观眼球不可见,病理学检查可见内有发育 异常的小眼球; 虹膜异常者可见瞳孔偏大,明显偏离中心位置,偏向位置不定,对光无反射,病理学观察可见虹膜晶 状体粘连、虹膜缺损,严重者伴有视网膜异常等。结论 本实验成功培育了 3 例眼部突变表型小鼠,为人类相关疾 病的研究提供良好的材料。     

Angiogenesis in cancer and other diseases

Pathological angiogenesis is a hallmark of cancer and various ischaemic and inflammatory diseases. Concentrated efforts in this area of research are leading to the discovery of a growing number of pro- and anti-angiogenic molecules, some of which are already in clinical trials. The complex interactions among these molecules and how they affect vascular structure and function in different environments are now beginning to be elucidated. This integrated understanding is leading to the development of a number of exciting and bold approaches to treat cancer and other diseases. But owing to several unanswered questions, caution is needed.     

铜代谢紊乱对脑神经系统疾病的影响

铜是维持哺乳动物正常生理活动的微量元素,体内铜的含量与细胞呼吸、自由基防御、血管生成、神经细胞和机体的生长发育有密切关系,它还参与脑生理活动的调节.体内铜的含量、转运及分布都受到精密的调控,铜元素缺乏和过量时都严重影响机体健康.铜及铜相关酶生物系统中对生命过程有着重要的作用,其代谢异常往往会导致严重的神经症状和神经系统疾病.本文将综述铜代谢紊乱对脑神经系统疾病的影响.     

Synthetic analogues of fumagillin that inhibit angiogenesis and suppress tumour growth.

Neovascularization is critical for the growth of tumours and is a dominant feature in a variety of angiogenic diseases such as diabetic retinopathy, haemangiomas, arthritis and psoriasis. Recognition of the potential therapeutic benefit of controlling unabated capillary growth has led to a search for safe and effective angiogenesis inhibitors. We report here the synthesis of a family of novel inhibitors that are analogues of fumagillin, a naturally secreted antibiotic of Aspergillus fumigatus fresenius. We first isolated this fungus from a contaminated culture of capillary endothelial cells. Purified fumagillin inhibited endothelial cell proliferation in vitro and tumour-induced angiogenesis in vivo; it also inhibited tumour growth in mice, but prolonged administration was limited because it caused severe weight loss. Synthesis of fumagillin analogues yielded potent angiogenesis inhibitors ('angioinhibins') which suppress the growth of a wide variety of tumours with relatively few side-effects.     

IL-17在胃癌组织中的表达及其与微血管生成的相关性分析

为探讨胃癌患者肿瘤组织中IL-17的表达及其与微血管生成的关系,采用免疫组织化学染色检测IL-17在30例胃癌患者石蜡包埋肿瘤组织中的表达,分析IL-17阳性细胞的形态学特征;对上述胃癌组织进行CD34免疫组织化学染色,计数微血管生成;统计学分析胃癌组织中IL-17表达与微血管生成的关系。结果在人类胃癌组织中IL-17阳性细胞多呈淋巴细胞样特征,且其表达水平较高组微血管生成增多。说明胃癌组织中IL-17表达水平与微血管生成正相关。     

Targeted misexpression of a Drosophila opsin gene leads to altered visual function

Drosophila mutants transformed with a chimaeric gene that expresses the ocellar visual pigment in the major class of photoreceptor cells of the retina were used to investigate the properties of this minor pigment. The photoreceptor cells in which this opsin was misexpressed showed new spectral characteristics and physiology.     

视网膜振荡电位异常改变的临床分析

目的 对多种眼科疾病进行视网膜振荡电位(Ops)的检测及分析，探讨了视网膜振荡电位在眼科多种疾病的诊断及评估方面的临床意义。方法对261例412只眼进行Ops检测，对Ops的检测结果进行分析。结果发现糖尿病及糖尿病性视网膜病变、视网膜中央及分支静脉阻塞、高度近视及高度近视性视网膜病变、视网膜色素变性等均有不同程度的Ops总幅值改变。同时发现，老年性黄斑变性、老年性白内障、弱视、中心性浆液性视网膜脉络膜病变及轻、中度近视者，则Ops总幅值无明显改变。结论视网膜Ops的检测是临床眼科诊断及评估的有力手段。     

雌激素刺激脑血管内皮细胞分泌VEGF及新血管形成

为了探讨雌激素对脑血管形成的影响,本实验利用体外培养的种植在matrigel上的脑血管内皮细胞作为模型,对雌激素刺激形成的新生血管进行照相记录,计算机软件分析.结果显示:雌激素呈剂量依赖性地刺激培养的脑血管内皮细胞分泌VEGF和新血管形成,而雄激素对培养的脑血管内皮细胞新血管形成没有任何影响;VEGF可以显著地促进内皮细胞新血管的形成.以上结果说明,雌激素通过刺激VEGF的分泌作用进而促进脑血管内皮细胞新血管的形成.     

Surface mechanics mediate pattern formation in the developing retina

Pattern formation of biological structures involves organizing different types of cells into a spatial configuration. In this study, we investigate the physical basis of biological patterning of the Drosophila retina in vivo. We demonstrate that E- and N-cadherins mediate apical adhesion between retina epithelial cells. Differential expression of N-cadherin within a sub-group of retinal cells (cone cells) causes them to form an overall shape that minimizes their surface contact with surrounding cells. The cells within this group, in both normal and experimentally manipulated conditions, pack together in the same way as soap bubbles do. The shaping of the cone cell group and packing of its components precisely imitate the physical tendency for surfaces to be minimized. Thus, simple patterned expression of N-cadherin results in a complex spatial pattern of cells owing to cellular surface mechanics.     

Expression and function of a new angiogenic factor AA98 target molecule at the maternal-embryonic boundary of rhesus monkey

The target molecule of monoclonal antibody AA98 (AA for short) is a new vascular endothelial cell related factor and plays a role in angiogenesis as indicated by the previous data. To investigate its role in angiogenesis and placentation in primate, we examined its expression in the implantation sites on D17, 19, 28 and 34 of gestation in rhesus monkey by immunohistochemistry and Western immunoblot. Western blot analysis showed that the primary antibody used in this study was specific for its epitope. AA protein was mainly expressed in small blood vessels and in some cytotrophoblast cells. The AA staining was found mainly in the endothelial cells and vascular small muscle. This observation supported the AA‘s role in angiogenesis. AA was spatio-temporarily expressed in cytotrophoblasts: weak in proliferating trophoblast within cell column and endovascular trophoblast, strong in trophoblastic subpopulation within the basal plate and vascular trophoblast; AA staining within the basal plate was down-regulated during early placentation. The shift of AA98 expression in extravillous trophoblasts suggestes a role of this new factor during the course of cytotrophoblast metastasis and spiral artery remodeling. The spatio-temporarily expression indicats that AA98 could be also used as a trophoblast cellular marker to characterize the acquisition of a vascular endothelial and invasive phenotype.     

Dll4 signalling through Notch1 regulates formation of tip cells during angiogenesis

In sprouting angiogenesis, specialized endothelial tip cells lead the outgrowth of blood-vessel sprouts towards gradients of vascular endothelial growth factor (VEGF)-A. VEGF-A is also essential for the induction of endothelial tip cells, but it is not known how single tip cells are selected to lead each vessel sprout, and how tip-cell numbers are determined. Here we present evidence that delta-like 4 (Dll4)-Notch1 signalling regulates the formation of appropriate numbers of tip cells to control vessel sprouting and branching in the mouse retina. We show that inhibition of Notch signalling using gamma-secretase inhibitors, genetic inactivation of one allele of the endothelial Notch ligand Dll4, or endothelial-specific genetic deletion of Notch1, all promote increased numbers of tip cells. Conversely, activation of Notch by a soluble jagged1 peptide leads to fewer tip cells and vessel branches. Dll4 and reporters of Notch signalling are distributed in a mosaic pattern among endothelial cells of actively sprouting retinal vessels. At this location, Notch1-deleted endothelial cells preferentially assume tip-cell characteristics. Together, our results suggest that Dll4-Notch1 signalling between the endothelial cells within the angiogenic sprout serves to restrict tip-cell formation in response to VEGF, thereby establishing the adequate ratio between tip and stalk cells required for correct sprouting and branching patterns. This model offers an explanation for the dose-dependency and haploinsufficiency of the Dll4 gene, and indicates that modulators of Dll4 or Notch signalling, such as gamma-secretase inhibitors developed for Alzheimer's disease, might find usage as pharmacological regulators of angiogenesis.     

Molecular mechanisms and clinical applications of angiogenesis

Blood vessels deliver oxygen and nutrients to every part of the body, but also nourish diseases such as cancer. Over the past decade, our understanding of the molecular mechanisms of angiogenesis (blood vessel growth) has increased at an explosive rate and has led to the approval of anti-angiogenic drugs for cancer and eye diseases. So far, hundreds of thousands of patients have benefited from blockers of the angiogenic protein vascular endothelial growth factor, but limited efficacy and resistance remain outstanding problems. Recent preclinical and clinical studies have shown new molecular targets and principles, which may provide avenues for improving the therapeutic benefit from anti-angiogenic strategies.     

Therapeutic angiogenesis induced by human hepatocyte growth factor (HGF) gene in rat myocardial ischemia models

Coronary arteriosclerotic cardiopathy is also named myocardial ischemia, which severely threatens humanhealth. Following the economic development and change of life style in China, population blood pressure, weight index and serum cholesterol level all rise. This prophesies incidence rate of coronary arteriosclerotic cardiopathy and stroke will increase year by year. Angioplasty and surgical bypass, the primary interventional therapies for these in-dividuals, are temporally limited by the prob…     

GAP-43在锦鲤荒漠沙蜥和雉鸡视网膜内分布的免疫组化研究

GAP-43具有多种功能,主要与神经元轴突的生长、再生、神经递质的释放及膜泡的吞噬有关.本研究用免疫组织化学的方法观察了正常成年锦鲤、荒漠沙蜥和雉鸡视网膜内GAP-43分布.结果显示GAP-43主要分布在内网层,另外,在内核层、外网层、光感受器细胞层因动物不同也呈现不同的分布特点,而在节细胞层中3种动物均未发现GAP-43阳性染色.在锦鲤视网膜中GAP-43主要分布在内网层和内核层的无长突细胞;在荒漠沙蜥视网膜中GAP-43主要分布在内网层和外网层,在雉鸡视网膜中GAP-43主要分布在内网层、外网层、外核层和光感受器外节,其中在雉鸡视网膜外核层和光感受器外节中发现阳性分布是在脊椎动物此层发现GAP-43的首次报道.     

转录因子Bach1在心血管疾病中的研究进展

转录因子Bach1(BTB-CNC同源体1, BTB and CNC homology 1)是一种转录抑制因子, 广泛存在于哺乳动物的各种组织中. Bach1能抑制血红素环氧化酶-1(heme oxygenase-1, HO-1)等抗氧化基因的表达, 参与氧化应激反应. Bach1基因敲除对心肌缺血再灌注损伤有保护作用. 近期研究发现Bach1能抑制Wnt/b-catenin信号通路和小鼠缺血下肢血管新生. 主要就Bach1在心血管疾病中的研究进展进行简要综述.     

Interactions between enkephalin and GABA in avian retina

In addition to conventional neurotransmitters such as acetylcholine, dopamine, glycine and gamma-aminobutyric acid (GABA), a number of peptide-immunoreactive substances have recently been localized in the vertebrate retina. The functional roles of these retinal peptides and their interactions with conventional neurotransmitters are largely unknown. We have previously shown that exogenous opiates affect both the release of GABA and the firing patterns of ganglion cells in the goldfish retina, and we have now begun a systematic characterization of the opioid pathways in the chicken retina, because, among vertebrate retinas, avian retinas contain the highest concentration of enkephalins. Monoclonal antibodies specific for enkephalin have been used to demonstrate that a subpopulation of enkephalin-containing amacrine cells exists in the chicken retina. This retina also synthesizes Met-enkephalin and releases it on cell depolarization. The enkephalin-induced inhibition of GABA release in goldfish retina led us to examine whether similar interactions occur in chicken, and if so, whether enkephalins and GABA coexist in the same amacrine cells. Our results, presented here, indicate that exogenous enkephalins do indeed inhibit GABA release in the chicken retina. Surprisingly, we found that although some amacrine cells contain both enkephalin and GABA, others contain only one or the other.     

中药对肿瘤组织血管生成的影响

恶性肿瘤的无限制侵袭性生长及其转移依赖于血管生成（angiogenesis)抑制血管生成是不同于常规的肿瘤治疗新策略，肿瘤血管生成的过程是个多步骤过程，目前已进入临床的肿瘤血管生成抑制剂有数十种，许多有抗肿瘤活性的中药的有效成分，如：人参皂苷Rg^3、鲨鱼软骨生成抑因子（SCAIF）等可抑制肿瘤血管生成。