BACKGROUND & AIMS: :This article describes the effects of dexmedetomidine (DEX) - the active ingredient of medetomidine, which is the latest popular sedative for functional magnetic resonance imaging (fMRI) in rodents - on multiple unit activity, local field potential (LFP), cerebral blood flow (CBF), pial vessel diameter [indicative of cerebral blood volume (CBV)], and blood oxygenation level-dependent (BOLD) fMRI. These measurements were obtained from the rat somatosensory cortex during 10 s of forepaw stimulation. We found that the continuous intravascular systemic infusion of DEX (50 μg/kg/h, doses typically used in fMRI studies) caused epileptic activities, and that supplemental isoflurane (ISO) administration of ~0.3% helped to suppress the development of epileptic activities and maintained robust neuronal and hemodynamic responses for up to 3 h. Supplemental administration of N(2)O in addition to DEX nearly abolished hemodynamic responses even if neuronal activity remained. Under DEX + ISO anesthesia, spike firing rate and the delta power of LFP increased, whereas beta and gamma power decreased, as compared with ISO-only anesthesia. DEX administration caused pial arteries and veins to constrict nearly equally, resulting in decreases in baseline CBF and CBV. Evoked LFP and CBF responses to forepaw stimulation were largest at a frequency of 8-10 Hz, and a non-linear relationship was observed. Similarly, BOLD fMRI responses measured at 9.4 T were largest at a frequency of 10 Hz. Both pial arteries and veins dilated rapidly (artery, 32.2%; vein, 5.8%), and venous diameter returned to baseline slower than arterial diameter. These results will be useful for designing, conducting and interpreting fMRI experiments under DEX sedation.

背景与目标： : 本文介绍了右美托咪定 (DEX) -美托咪定的有效成分，美托咪定是啮齿动物功能磁共振成像 (fMRI) 的最新流行镇静剂-对多单位活动，局部场电位 (LFP)，脑血流 (CBF)，血管直径 [指示脑血容量 (CBV)] 和血氧水平依赖性 (BOLD) fMRI。这些测量值是在前爪刺激10 s期间从大鼠体感皮层获得的。我们发现，连续血管内全身输注DEX (50 μ g/kg/h，通常用于fMRI研究的剂量) 引起癫痫活动，并且〜0.3% 的补充异氟烷 (ISO) 给药有助于抑制癫痫活动的发展，并在长达3小时的时间内保持强大的神经元和血液动力学反应。除DEX外，补充施用N(2)O几乎消除了血液动力学反应，即使神经元活性仍然存在。在DEX ISO麻醉下，与仅ISO麻醉相比，尖峰放电速率和LFP的 δ 功率增加，而 β 和 γ 功率降低。DEX给药导致腰肌动脉和静脉几乎相等地收缩，导致基线CBF和CBV降低。诱发的LFP和CBF对前爪刺激的反应在8-10Hz的频率下最大，并且观察到非线性关系。类似地，在9.4 T处测量的BOLD fMRI响应在10 hz的频率下最大。腰肌动脉和静脉均迅速扩张 (动脉，32.2%; 静脉，5.8%)，并且静脉直径恢复到基线的速度比动脉直径慢。这些结果将有助于在DEX镇静下设计，进行和解释fMRI实验。

BACKGROUND & AIMS: :Menopausal hormone treatment (MHT) may limit progression of cardiovascular disease (CVD) but poses a thrombosis risk. To test targeted candidate gene variation for association with subclinical CVD defined by carotid artery intima-media thickness (CIMT) and coronary artery calcification (CAC), 610 women participating in the Kronos Early Estrogen Prevention Study (KEEPS), a clinical trial of MHT to prevent progression of CVD, were genotyped for 13,229 single nucleotide polymorphisms (SNPs) within 764 genes from anticoagulant, procoagulant, fibrinolytic, or innate immunity pathways. According to linear regression, proportion of European ancestry correlated negatively, but age at enrollment and pulse pressure correlated positively with CIMT. Adjusting for these variables, two SNPs, one on chromosome 2 for MAP4K4 gene (rs2236935, β = 0.037, P value = 2.36 × 10(-06)) and one on chromosome 5 for IL5 gene (rs739318, β = 0.051, P value = 5.02 × 10(-05)), associated positively with CIMT; two SNPs on chromosome 17 for CCL5 (rs4796119, β = -0.043, P value = 3.59 × 10(-05); rs2291299, β = -0.032, P value = 5.59 × 10(-05)) correlated negatively with CIMT; only rs2236935 remained significant after correcting for multiple testing. Using logistic regression, when we adjusted for waist circumference, two SNPs (rs11465886, IRAK2, chromosome 3, OR = 3.91, P value = 1.10 × 10(-04); and rs17751769, SERPINA1, chromosome 14, OR = 1.96, P value = 2.42 × 10(-04)) associated positively with a CAC score of >0 Agatston unit; one SNP (rs630014, ABO, OR = 0.51, P value = 2.51 × 10(-04)) associated negatively; none remained significant after correcting for multiple testing. Whether these SNPs associate with CIMT and CAC in women randomized to MHT remains to be determined.

背景与目标： : 更年期激素治疗 (MHT) 可能会限制心血管疾病 (CVD) 的进展，但会带来血栓形成的风险。为了测试与颈动脉内膜中层厚度 (CIMT) 和冠状动脉钙化 (CAC) 定义的亚临床CVD相关的靶向候选基因变异，610参加Kronos早期雌激素预防研究 (KEEPS) 的妇女，MHT预防CVD进展的临床试验，在抗凝剂，促凝剂，纤溶或先天免疫途径的764基因内对13,229单核苷酸多态性 (snp) 进行基因分型。根据线性回归，欧洲血统的比例呈负相关，但入学年龄和脉压与CIMT呈正相关。调整这些变量，两个snp，一个在2号染色体上的MAP4K4基因 (rs2236935，β = 0.037，p值 = 2.36 × 10(-06))，一个在5号染色体上的IL5基因 (rs739318，β = 0.051，p值 = 5.02 × 10(-05))，与CIMT呈正相关; CCL5 17号染色体上的两个snp (rs4796119，β = -0.043，p值 = 3.59 × 10(-05); rs2291299，β = -0.032，p值 = 5.59 × 10(-05)) 与CIMT呈负相关; 校正多重测试后，只有rs2236935仍然显著。使用逻辑回归，当我们调整腰围时，两个snp (rs11465886，IRAK2，3号染色体，OR = 3.91，p值 = 1.10 × 10(-04); 和rs17751769，SERPINA1，14号染色体，OR = 1.96，p值 = 2.42 × 10(-04)) 与> 0 Agatston单位的CAC评分呈正相关; 1个SNP (rs630014，ABO，OR = 0.51，p值 = 2.51 × 10(-04)) 呈负相关; 校正多重测试后无显著。这些snp是否与CIMT和CAC相关，在随机分配到MHT的女性中仍有待确定。

BACKGROUND & AIMS: :We are developing biocompatible small-calibre vascular substitutes based on polymeric scaffolds that incorporate cell-matrix signals to enhance vascular cell attachment and function. Our graft scaffold comprises an outer electrostatically spun porous polyurethane layer seeded with smooth muscle cells, and a luminal polycaprolactone layer for endothelial cell attachment. Vascular cell adhesion properties of three vascular elastic fibre molecules, tropoelastin, fibrillin-1 and fibulin-5, have been defined, and adhesion fragments optimized. These fragments are being used to coat the scaffolds to enhance luminal endothelial cell attachment, and to regulate smooth muscle cell attachment and function. Tropoelastin-based cell seeding materials are also being developed. In this way, vascular cell-matrix biology is enhancing graft design.

背景与目标： : 我们正在开发基于聚合物支架的生物相容性小口径血管替代物，该支架结合了细胞基质信号以增强血管细胞附着和功能。我们的移植支架包括一个外部静电纺丝多孔聚氨酯层，该层接种有平滑肌细胞，以及一个用于内皮细胞附着的内腔聚己内酯层。已经定义了三种血管弹性纤维分子 (原弹性蛋白，fibrillin-1和fibulin-5) 的血管细胞粘附特性，并优化了粘附片段。这些片段被用于涂覆支架，以增强管腔内皮细胞的附着，并调节平滑肌细胞的附着和功能。基于原弹性蛋白的细胞播种材料也正在开发中。通过这种方式，血管细胞基质生物学正在增强移植物的设计。

BACKGROUND & AIMS: OBJECTIVE:Ossification/calcification around the medial femoral condyle has been known as Pellegrini-Stieda (PS) disease for almost 100 years. Little attention has been given to magnetic resonance (MR) imaging characteristics. Our purpose is to demonstrate the anatomy in the medial femoral compartment and imaging findings of PS disease, determining the sites and patterns of ossification. DESIGN AND PATIENTS:In a cadaveric study seven specimens were dissected to show the anatomic relations of the tibial collateral ligament (TCL) and the tendon of the ischiocondylar part of the adductor magnus muscle, in the medial femoral epicondyle. In order to determine the nature of ossification/calcification in PS disease, MR imaging and radiographic findings in nine patients were analyzed by two observers with attention to the specific site, shape, and orientation of the ossification and its relationship to the tibial collateral ligament (TCL) and adductor magnus tendon. Available clinical history was recorded. A classification system addressing different sites and patterns of ossification was developed. RESULTS:The anatomic study showed that the TCL and the adductor magnus tendon insert at different sites in the medial femoral condyle and there is no continuation; however, some fibers of the posterior bundle of the TCL overlap the anterior aspect of the adductor magnus tendon. The imaging study showed that shape, orientation, and location of the abnormal calcification and ossification were similar on radiographic and MR imaging analysis. Ossification had an inferior orientation in six cases, a superior orientation in two cases, and both in one case. Four patterns of ossification were noted: (I) a beak-like appearance with an inferior orientation and femoral attachment was present in five cases; (II) a drop-like appearance with an inferior orientation, parallel to the femur, was evident in one case; (III) an elongated appearance with a superior orientation, parallel to the femur, was seen in two cases; and (IV) a beak-like appearance with an inferior and superior orientation, attached to the femur, was seen in one case. The ossification was present in the TCL in six cases, in the adductor magnus tendon in two cases, and in both in one case. The coronal plane was best in detecting and categorizing the ossification. CONCLUSION:Our data indicate that ossification in PS disease is not confined to the TCL but may also involve the adductor magnus tendon. In some cases, it can be related to the anatomic proximity (overlap) of the fibers of these two structures. PS disease should not be regarded as synonymous with ossification of the TCL. The ossification may be classified into four types. No clinical differences among these types appear to exist.

背景与目标：

BACKGROUND & AIMS: :We described seven patients with Streptococcus milleri group aortic (six patients) or vena cava (one patient) graft infection secondary to a vasculo-digestive fistula. Time between vascular graft setting and first clinical signs varied from eight months to more than thirteen years. Six patients had fever. Three patients presented with recurrent fever for more than nine months and in two of these cases, delay before diagnosis was long because repeated blood cultures were sterile. Three patients had abdominal pain and/or digestive haemorrhage. Abdominal CT-scan S. milleri was not contributive for the diagnosis in four patients. Streptococcus anginosus was isolated in four patients, Streptococcus constellatus in three patients. One patient died before surgical management. The other six patients were cured by a surgical management associated with a prolonged antibiotic (lactams) treatment. S. milleri group graft infections are rare (or misdiagnosed) while we found only 4 similar cases in the English medical literature. We conclude that a peri-prosthetic infection secondary to a digestive fistula must be insistently searched (and blood cultures must be repeated many times) in any patient with an aortic (or any other vascular) graft presenting prolonged or recurrent fever or acute digestive symptoms.

背景与目标： : 我们描述了7例继发于血管消化瘘的链球菌组主动脉 (6例) 或腔静脉 (1例) 移植物感染的患者。血管移植与首次临床体征之间的时间从8个月到13年以上不等。六名患者发热。三名患者出现反复发热超过9个月，其中两名患者由于反复的血液培养是无菌的，因此在诊断之前的延迟时间很长。三名患者患有腹痛和/或消化道出血。腹部ct扫描S. milleri对四名患者的诊断没有帮助。在4例患者中分离出aninosus链球菌，在3例患者中分离出constellatus链球菌。一名患者在手术治疗前死亡。其他六名患者通过长期抗生素 (lacams) 治疗的外科手术治愈。S. milleri组移植物感染很少见 (或误诊)，而我们在英国医学文献中仅发现4例类似病例。我们得出的结论是，对于任何主动脉 (或任何其他血管) 移植物出现长期或反复发烧或急性消化症状的患者，必须坚持检查继发于消化瘘的假体周围感染 (并且必须重复多次血液培养)。

BACKGROUND & AIMS: BACKGROUND:An impairment of cerebral microvessels is reported both in normal ageing and in senescence-associated processes, as well as in Alzheimer's disease (AD) and vascular dementia (VaD). The aim of this study was to explore cerebral hemodynamics by transcranial Doppler in VaD and AD, compared with age-matched control subjects. METHODS:Transcranial Doppler was investigated in all patients in the basal condition. Cerebral vasoreactivity to hyper- and hypocapnia was evaluated with CO2 mixture inhalation followed by hyperventilation. RESULTS:We studied 60 AD and 58 VaD patients and 62 nondemented controls. Both AD and VaD subjects showed lower flow velocities (FV) and higher pulsatility indices (PI) as compared with controls. Lower total vasomotor reactivity and lower response to hypercapnia were observed in the AD and VaD groups as compared with controls. AD and VaD patients did not show significant differences in FV, PI values or cerebral vasoreactivity. CONCLUSIONS:Reduced FV and increased PI with a significant vasoreactivity reduction in VaD and AD patients are indicators of impairment of cerebral microvasculature circulation in both diseases. The identification of vascular function impairment in all kinds of dementia could be of help in identifying patients who would thus benefit more from specific therapeutic approaches.

背景与目标：

BACKGROUND & AIMS: :Although the involvement of Rho proteins in the pathogenesis of vascular diseases is well studied, little is known about the role of their upstream regulators, the Rho guanine nucleotide exchange factors (RhoGEFs). Here, we sought to identify the RhoGEFs involved in monocyte chemotactic protein 1 (MCP1)-induced vascular wall remodeling. We found that, among the RhoGEFs tested, MCP1 induced tyrosine phosphorylation of p115 RhoGEF but not of PDZ RhoGEF or leukemia-associated RhoGEF in human aortic smooth muscle cells (HASMCs). Moreover, p115 RhoGEF inhibition suppressed MCP1-induced HASMC migration and proliferation. Consistent with these observations, balloon injury (BI) induced p115 RhoGEF tyrosine phosphorylation in rat common carotid arteries, and siRNA-mediated down-regulation of its levels substantially attenuated BI-induced smooth muscle cell migration and proliferation, resulting in reduced neointima formation. Furthermore, depletion of p115 RhoGEF levels also abrogated MCP1- or BI-induced Rac1-NFATc1-cyclin D1-CDK6-PKN1-CDK4-PAK1 signaling, which, as we reported previously, is involved in vascular wall remodeling. Our findings also show that protein kinase N1 (PKN1) downstream of Rac1-cyclin D1/CDK6 and upstream of CDK4-PAK1 in the p115 RhoGEF-Rac1-NFATc1-cyclin D1-CDK6-PKN1-CDK4-PAK1 signaling axis is involved in the modulation of vascular wall remodeling. Of note, we also observed that CCR2-Gi/o-Fyn signaling mediates MCP1-induced p115 RhoGEF and Rac1 GTPase activation. These findings suggest that p115 RhoGEF is critical for MCP1-induced HASMC migration and proliferation in vitro and for injury-induced neointima formation in vivo by modulating Rac1-NFATc1-cyclin D1-CDK6-PKN1-CDK4-PAK1 signaling.

背景与目标： : 尽管对Rho蛋白参与血管疾病的发病机理进行了很好的研究，但对其上游调节剂Rho鸟嘌呤核苷酸交换因子 (rhoiefs) 的作用知之甚少。在这里，我们试图鉴定参与单核细胞趋化蛋白1 (MCP1) 诱导的血管壁重塑的RhoGEFs。我们发现，在测试的RhoGEFs中，MCP1诱导人主动脉平滑肌细胞 (HASMCs) 中p115 RhoGEF的酪氨酸磷酸化，但不诱导PDZ RhoGEF或与白血病相关的RhoGEF的酪氨酸磷酸化。此外，p115 rhoge抑制抑制了MCP1-induced HASMC的迁移和增殖。与这些观察结果一致，球囊损伤 (BI) 在大鼠颈总动脉中诱导了p115 RhoGEF酪氨酸磷酸化，而siRNA介导的其水平下调大大减弱了BI诱导的平滑肌细胞迁移和增殖，从而减少了新内膜的形成。此外，p115 RhoGEF水平的耗竭也消除了MCP1或BI诱导的Rac1-NFATc1-cyclin D1-CDK6-PKN1-CDK4-PAK1信号传导，正如我们先前报道的那样，这与血管壁重塑有关。我们的发现还表明，Rac1-cyclin D1/CDK6下游和p115 RhoGEF-Rac1-NFATc1-cyclin D1-CDK6-PKN1-CDK4-PAK1信号轴CDK4-PAK1上游的蛋白激酶N1 (PKN1) 参与了血管壁重塑的调节。值得注意的是，我们还观察到CCR2-Gi/o-Fyn信号介导MCP1-induced p115 rhoge和rac1gtpase激活。这些发现表明，p115 RhoGEF对于MCP1-induced HASMC在体外迁移和增殖以及通过调节Rac1-NFATc1-cyclin D1-CDK6-PKN1-CDK4-PAK1信号传导在体内损伤诱导的新内膜形成至关重要。

BACKGROUND & AIMS: BACKGROUND/AIMS:Cardiovascular disease (CVD) is increased in chronic kidney disease (CKD), and contributed to by the CKD-mineral bone disorder (CKD-MBD). CKD-MBD begins in early CKD and its vascular manifestations begin with vascular stiffness proceeding to increased carotid artery intima-media thickness (cIMT) and vascular calcification (VC). Phosphorus is associated with this progression and is considered a CVD risk factor in CKD. We hypothesized that modifying phosphorus balance with lanthanum carbonate (LaCO3) in early CKD would not produce hypophosphatemia and may affect vascular manifestations of CKD-MBD. METHODS:We randomized 38 subjects with normophosphatemic stage 3 CKD to a fixed dose of LaCO3 or matching placebo without adjusting dietary phosphorus in a 12-month randomized, double-blind, pilot and feasibility study. The primary outcome was the change in serum phosphorus. Secondary outcomes were changes in measures of phosphate homeostasis and vascular stiffness assessed by carotid-femoral pulse wave velocity (PWV), cIMT and VC over 12 months. RESULTS:There were no statistically significant differences between LaCO3 and placebo with respect to the change in serum phosphorus, urinary phosphorus, tubular reabsorption of phosphorus, PWV, cIMT, or VC. Biomarkers of the early CKD-MBD such as plasma fibroblast growth factor-23, Dickkopf-related protein 1 (DKK1), and sclerostin were increased 2- to 3-fold at baseline, but were not affected by LaCO3. CONCLUSION:Twelve months of LaCO3 had no effect on serum phosphorus and did not alter phosphate homeostasis, PWV, cIMT, VC, or biomarkers of CKD-MBD.

背景与目标：

BACKGROUND & AIMS: :This study investigates whether salivary tumours with different morphology and evolution also differ in terms of neovascularization and VEGF expression and the prognostic value of the results. Surgical specimens from 45 patients - 8 pleomorphic adenomas (PA), 7 Warthin tumours (WT), 5 basal cell adenomas (BA), 6 carcinomas ex-pleomorphic adenoma (CEPA), 6 mucoepidermoid carcinomas (MEC), 5 acinic cell carcinomas (AC), 4 adenoid cystic carcinomas (ACC) and 4 adenocarcinomas not otherwise specified (ADK NOS) - were immunostained. In malignant salivary tumours, the following mean microvascular density (MVD) values were recorded (± SD = Standard Deviation): 27.61 (SD ± 2.27) in cases with CEPA, 27.08 (DS ± 7.81) in AC and 32.93 (SD ± 7.76) in ADK NOS, with lower values for MEC 24.31(SD ± 2.88) and for ACC 22.13 (SD ± 5.44). For benign tumours, an MVD of 35.71 (SD ± 2.09) was recorded in WT and lower average values in PA (MVD = 14.84; SD ± 4.86) and in BA (MVD = 23.96; SD ± 9.13). MVD did not correlate with the investigated clinicopathological parameters. The VEGF expression is significantly more important (p = 0.001) in malignant salivary tumours as compared with benign ones. The VEGF expression and the microvascularization in salivary gland tumours are important elements to be considered when formulating a diagnosis and assessing case evolutions in patients with such tumours.

背景与目标： : 本研究调查了具有不同形态和演变的唾液肿瘤在新生血管形成和VEGF表达以及结果的预后价值方面是否也存在差异。45例患者的手术标本-8例多形性腺瘤 (PA)，7例Warthin肿瘤 (WT)，5例基底细胞腺瘤 (BA)，6例非多形性腺瘤 (CEPA)，6例粘液表皮样癌 (MEC)，5例腺泡细胞癌 (AC)，免疫染色了4例腺样囊性癌 (ACC) 和4例未指明的腺癌 (ADK NOS)。在恶性唾液肿瘤中，记录了以下平均微血管密度 (MVD) 值 (± SD = 标准差): CEPA病例为27.61 (SD ± 2.27)，AC为27.08 (DS ± 7.81)，ADK NOS为32.93 (SD ± 7.76)，MEC 24.31(SD ± 2.88) 和ACC 22.13 (SD ± 5.44) 的值较低。对于良性肿瘤，在WT中记录了35.71的MVD (SD ± 2.09)，在PA (MVD = 14.84; SD ± 4.86) 和BA (MVD = 23.96; SD ± 9.13) 中记录了较低的平均值。MVD与所研究的临床病理参数无关。与良性唾液肿瘤相比，恶性唾液肿瘤中VEGF表达明显更重要 (p = 0.001)。涎腺肿瘤中的VEGF表达和微血管形成是制定诊断和评估此类肿瘤患者病例演变时要考虑的重要因素。

BACKGROUND & AIMS: :Cerebrovascular disease (CVD) and amyloid burden are the most frequent pathologies in subjects with cognitive impairment. However, the relationship between CVD, amyloid burden, and cognition are largely unknown. We aimed to evaluate whether CVD (lacunes, white matter hyperintensities, and microbleeds) and amyloid burden (Pittsburgh compound B [PiB] retention ratio) contribute to cognitive impairment independently or interactively. We recruited 136 patients with subcortical vascular cognitive impairment who underwent magnetic resonance imaging, PiB-positron emission tomography, and neuropsychological testing. The number of lacunes was associated with memory, frontal dysfunctions, and disease severity. The volume of white matter hyperintensities and the PiB retention ratio were associated only with memory dysfunction. There was no direct correlation between CVD markers and PiB retention ratio except that the number of lacunes was negatively correlated with the PiB retention ratio. In addition, there were no interactive effects of CVD and PiB retention ratio on cognition. Our findings suggest that CVD and amyloid burden contribute independently and not interactively to specific patterns of cognitive dysfunction in patients with subcortical vascular cognitive impairment.

背景与目标： : 脑血管疾病 (CVD) 和淀粉样蛋白负担是认知障碍患者最常见的病理。然而，CVD，淀粉样蛋白负担和认知之间的关系在很大程度上尚不清楚。我们旨在评估CVD (腔隙，白质高信号和微出血) 和淀粉样蛋白负荷 (匹兹堡化合物B [PiB] 保留比) 是否独立或交互地导致认知障碍。我们招募了136名皮质下血管性认知障碍患者，他们接受了磁共振成像，PiB-正电子发射断层扫描和神经心理学测试。腔隙的数量与记忆，额叶功能障碍和疾病严重程度有关。白质高信号的体积和PiB保留率仅与记忆功能障碍有关。CVD标记与PiB保留率之间没有直接相关性，只是腔隙数与PiB保留率呈负相关。此外，CVD和PiB保留率对认知没有交互作用。我们的发现表明，在皮质下血管性认知障碍患者中，CVD和淀粉样蛋白负担独立且非交互作用地促进了认知功能障碍的特定模式。

BACKGROUND & AIMS: :Chemotherapy-induced alopecia is a major problem in clinical oncology. Doxorubicin, a widely used cancer chemotherapy drug, induces disruption of the hair cycle and subsequent alopecia. We show in this report that doxorubicin causes disruption of the hair-follicle-associated blood vessel network resulting in a greatly reduced density of these blood vessels. Dystrophic hair follicles were also observed with abnormal melanogenesis in the mice treated with doxorubicin. Visualization of the effect of doxorubicin on hair-follicle angiogenesis was made possible by the use of transgenic mice in which green fluorescent protein was driven by regulatory elements of the nestin gene (ND-GFP). In these transgenic mice, the hair-follicle stem cells and the follicle structure as well as the blood vessels associated with the hair follicles express ND-GFP. The hair-follicle stem cells did not appear to be affected by doxorubicin, which may explain why hair regrows after chemotherapy. These results suggest that inhibition of hair-follicle-associated angiogenesis by doxorubicin may be an important factor in hair-follicle dystrophy associated with chemotherapy-induced alopecia. The ND-GFP mouse model is thus useful for the study of the role of angiogenesis in the hair-follicle cycle and the effect of drugs on processes associated with chemotherapy-induced alopecia.

背景与目标： : 化疗引起的脱发是临床肿瘤学的主要问题。阿霉素是一种广泛使用的癌症化疗药物，可引起头发周期的破坏和随后的脱发。我们在本报告中显示，阿霉素会导致毛囊相关血管网络的破坏，从而导致这些血管的密度大大降低。在用阿霉素治疗的小鼠中，还观察到营养不良的毛囊具有异常的黑素生成。通过使用转基因小鼠，可以可视化阿霉素对毛囊血管生成的作用，其中绿色荧光蛋白由巢蛋白基因 (ndi-GFP) 的调控元件驱动。在这些转基因小鼠中，毛囊干细胞和毛囊结构以及与毛囊相关的血管表达ND-GFP。毛囊干细胞似乎不受阿霉素的影响，这可以解释为什么化疗后头发再生。这些结果表明，阿霉素抑制毛囊相关的血管生成可能是与化疗引起的脱发相关的毛囊营养不良的重要因素。因此，ND-GFP小鼠模型可用于研究血管生成在毛囊周期中的作用以及药物对与化学疗法诱导的脱发相关的过程的影响。

BACKGROUND & AIMS: :Somatic gene transfer continues to have potential for the study and therapy of cardiovascular disease. We have developed a modular, self-assembling, nonviral system consisting of Lipofectin, integrin-targeting peptides, and plasmid DNA (LID) and we have applied this to a model of vascular injury, rat carotid angioplasty. Marker gene studies identified transfection of adventitial cells after vector delivery to that layer. Human tissue inhibitor of metalloproteinase-1 (hTIMP-1) was tested as a therapeutic gene product after direct application to the exposed adventitial layer. Vascular LID.hTIMP-1 transfection was confirmed by polymerase chain reaction and gene expression by immunohistochemistry at 7 days. Neointimal areas were 0.160 +/- 0.078 and 0.225 +/- 0.052 mm(2) for LID.hTIMP-1-transfected (n = 14) and LID.pCI-transfected (n = 12) vessels, respectively, at 14 days, and 0.116 +/- 0.068 mm(2) (n = 14) and 0.194 +/- 0.095 mm(2) (n = 14), respectively, at 28 days, representing a 29 and 40% reduction in neointimal hyperplasia at 14 and 28 days, respectively, after balloon dilatation. Neointima-to-media ratios were similarly reduced. In addition, expansile remodeling after balloon injury was inhibited at 14 days, the area within the external elastic lamina being 0.50 +/- 0.02 and 0.61 +/- 0.02 mm(2) in LID.hTIMP-1- and LID.pCI-transfected arteries, respectively (p < 0.0005). We have demonstrated an effective system of therapeutic gene transfer, particularly targeting the arterial adventitia, where transfer of genes involved in matrix remodeling and cell migration may be useful.

背景与目标： : 体细胞基因转移继续具有研究和治疗心血管疾病的潜力。我们已经开发了一种模块化，自组装的非病毒系统，该系统由Lipofectin，整联蛋白靶向肽和质粒DNA (LID) 组成，并将其应用于血管损伤模型大鼠颈动脉血管成形术。标记基因研究确定了载体递送到该层后外膜细胞的转染。在直接施用于暴露的外膜层后，测试了metalloproteinase-1的人组织抑制剂 (hTIMP-1) 作为治疗性基因产物。血管盖。在第7天，通过聚合酶链反应和免疫组织化学证实hTIMP-1转染。在第14天，LID.hTIMP-1-transfected (n = 14) 和LID.pCI转染 (n = 12) 血管的内膜面积分别为0.160 +/- 0.078和0.225 +/-0.052毫米 (2)，和0.116 +/-0.068毫米 (2) (n = 14) 和0.194 +/-0.095毫米 (2) (n = 14) 分别在第28天，分别表示在第14天和第28天新生内膜增生减少29和40%，球囊扩张后。新内膜与中膜比率也同样降低。此外，在第14天，球囊损伤后的扩张重塑受到抑制，LID.hTIMP-1和LID.pCI转染的动脉中，外部弹性层内的面积分别为0.50 +/- 0.02和0.61 +/-0.02毫米 (2) (p <0.0005)。我们已经证明了一种有效的治疗性基因转移系统，尤其是针对动脉外膜，其中涉及基质重塑和细胞迁移的基因转移可能是有用的。

BACKGROUND & AIMS: OBJECTIVES:Angiogenesis, the process of formation of blood vessels, is essential for many physiological as well as pathological processes. It has been shown that human adipose tissue contains a population of non-characterized cells, called stromal-vascular fraction (SVF) cells, which are able to differentiate into several lineages. The aim of this study was to determine conditions for promoting differentiation of human adipose tissue progenitors towards endothelial cells, as well as to show that SVF cells cooperate with differentiated endothelium in capillary network formation. MATERIALS AND METHODS:Stromal vascular fraction cells were isolated according to modified Hauner's method and after adaptation they were cultured in pro-angiogenic or pro-adipogenic medium. Cells were characterized by presence of surface antigens by flow cytometry, and by expression of genes characteristic for endothelial cells or for adipocytes, quantitative real-time polymerase chain reaction. A number of tests were performed to verify their differentiation. RESULTS:Differentiation of human SVF cells towards endothelium was stimulated by the presence of serum and absence of adipogenic factors, documented by the pattern of gene expression as well as different functional in vitro assays. SVF cells were found to work together with human umbilical vein endothelial cells to form capillary networks. CONCLUSIONS:Here, we show that differentiation of SVF cells to endothelial cells or adipocyte-like cells depended on the medium used. Our work provides a clear model for analysing the differentiation capacity of SVF cells.

背景与目标：

BACKGROUND & AIMS: OBJECTIVES:To determine whether vascular endothelial growth factor A (VEGF) and its receptors are expressed during bladder development in mice when capillaries are forming, and whether exogenous VEGF might enhance the growth of endothelia and other types of bladder cells, using an embryonic organ-culture model. MATERIALS AND METHODS:Whole bladders from wild-type mice, at embryonic day (E) 14, were grown in serum-free organ culture in an air/5% CO2 atmosphere; some cultures were supplemented with VEGF and/or with VEGF receptor 1/Fc chimera (VEGFR1/Fc), which blocks VEGF bioactivity. Organs were harvested after 6 days and the expression of VEGF and related molecules assessed using immunohistochemistry. RESULTS:VEGF, VEGFR1 and VEGFR2 positive cells were immunodetected in E14 and E18 bladders. Exogenous VEGF increased whole-organ growth, as assessed by explant areas, total cell numbers, DNA and protein content; proliferation was enhanced, and apoptosis decreased, in urothelium and surrounding tissues. VEGF also increased the proportions of cells expressing endothelial (CD31) and smooth muscle (alpha smooth muscle actin) markers. VEGFR1/Fc blocked the growth-enhancing effects of exogenous VEGF. CONCLUSIONS:In organ culture, exogenous VEGF not only stimulated embryonic bladder endothelial cells but also strikingly enhanced the growth of the whole organ. Whether the effects of VEGF on diverse bladder cell populations are direct or indirect requires further investigation. The finding that VEGF protein is present in embryonic bladders in vivo raises the possibility that it has similar actions during normal development. The results also illuminate the pathobiology of certain bladder diseases in which VEGF levels have been shown to be increased.

背景与目标：

BACKGROUND & AIMS: :Previous in vivo and in vitro studies have shown that Akt1 serves as a crucial regulator of vascular maturation, extracellular matrix composition, and angiogenesis in tumors. Hence, we hypothesized that Akt1 may be necessary for other angiogenesis-dependent processes, including wound healing. Using Akt1 (-/-) and Akt2 (-/-) mice, we demonstrate that deficiency of Akt1, but not Akt2, results in impaired assembly of collagen in skin wounds and around the blood vessels. Wounds in Akt1 (-/-) mice, but not in Akt2 (-/-) mice, were characterized by reduced vascular area as well as impaired vascular maturation as evidenced by reduced smooth muscle cell recruitment. Expression level of a major angiogenic growth factor, VEGF, was significantly lower in wound tissues of Akt1 (-/-) mice as compared to WT. However, despite the impaired collagen assembly and reduced angiogenesis in Akt1 (-/-) wounds, no significant difference in migration of fibroblasts into the wound area was observed between WT and Akt1 (-/-) mice. Importantly, the dynamics of wound closure were similar between WT, Akt1 (-/-), and Akt2 (-/-) mice. Thus, it appears that although the lack of Akt1 impairs VEGF expression, wound angiogenesis, and subsequent maturation of vasculature, it has no effect on the wound closure. These findings may have clinical applications for the improvement of treatment procedures with reported history of wound healing complications.

背景与目标： : 先前的体内和体外研究表明，Akt1是肿瘤中血管成熟，细胞外基质组成和血管生成的关键调节剂。因此，我们假设Akt1可能是其他血管生成依赖性过程 (包括伤口愈合) 所必需的。使用Akt1 (-/-) 和Akt2 (-/-) 小鼠，我们证明Akt1的缺乏而不是Akt2的缺乏会导致皮肤伤口和血管周围胶原蛋白的组装受损。Akt1 (-/-) 小鼠的伤口，但Akt2 (-/-) 小鼠的伤口的特征是血管面积减少以及血管成熟受损，平滑肌细胞募集减少证明了这一点。与WT相比，Akt1 (-/-) 小鼠伤口组织中主要血管生成生长因子VEGF的表达水平显着降低。然而，尽管在Akt1 (-/-) 伤口中胶原蛋白组装受损且血管生成减少，但在WT和Akt1 (-/-) 小鼠之间未观察到成纤维细胞向伤口区域迁移的显着差异。重要的是，WT，Akt1 (-/-) 和Akt2 (-/-) 小鼠之间的伤口闭合动力学相似。因此，尽管缺乏Akt1会损害VEGF的表达，伤口血管生成以及随后的脉管系统成熟，但它对伤口闭合没有影响。这些发现可能具有临床应用，可用于改善伤口愈合并发症史的治疗程序。