Role of the NF-kB/parkin/vegfr-1 pathway associated with hypoxic-ischemic insult in germinal matrix samples of newborn infants / O papel da via NF-kB/parkin/vegfr-1 associada à agressão hipóxico-isquêmica em amostras de matriz germinativa de pacientes recém-nascidos

Abstract Objective: Given the high proliferative activity of germinal matrix and its direct correlation with hypoxemia, it is necessary to investigate the possible molecular regulation pathways, to understand the existing clinical relationship between the hypoxic-ischemic insult and the biomarkers NF-kB, AKT-3, Parkin, TRK-C and VEGFR-1. Methods: A hundred and eighteen germinal matrix samples of the central nervous system of patients who died in the first 28 days of life were submitted to histological and immunohistochemistry analysis to identify the tissue immunoexpression of those biomarkers related to asphyxia, prematurity, and death events within 24h. Results: A significantly increased tissue immunoexpression of NF-kB, AKT-3 and Parkin was observed in the germinal matrix of preterm infants. In addition, significantly decreased tissue immunoexpression of VEGFR-1 and NF-kB was observed in patients who experienced asphyxia followed by death within 24 hours. Conclusions: The results suggest a direct involvement between the hypoxic-ischemic insult and NF-kB and VEGFR-1 markers since a decreased immunoexpression of these biomarkers was observed in asphyxiated patients. Furthermore, it is suggested that there was not enough time for VEGFR-1 to be transcribed, translated and expressed on the surface of the plasma membrane. This temporality can be observed in the relationship between NF-kB expression and the survival time of individuals who died within 24 hours, suggesting that this factor is essential for the production of VEGFR-1 and, therefore, to carry out the necessary remodeling effect to neovascularize the affected region.

RESUMO Objetivo: Dada a alta atividade proliferativa da matriz germinativa e sua correlação direta com a hipoxemia, é necessário investigar as possíveis vias de regulação molecular para entender a relação clínica existente entre o insulto hipóxico-isquêmico e os biomarcadores NF-kB, AKT -3, Parkina, TRK-C e VEGFR-1. Métodos: Cento e dezoito amostras de matriz germinativa do sistema nervoso central de pacientes que faleceram nos primeiros 28 dias de vida foram submetidas a análise histológica e imuno-histoquímica para identificar a imunoexpressão tecidual desses biomarcadores relacionados a eventos de asfixia, prematuridade e óbito em 24 horas. Resultados: Observou-se uma imunoexpressão tecidual significativamente aumentada de NF-kB, AKT-3 e Parkin na matriz germinativa de prematuros. Além disso, constatou-se uma imunoexpressão tecidual significativamente diminuída de VEGFR-1 e de NF-kB em pacientes que apresentaram asfixia seguida de morte em 24 horas. Conclusões: Os resultados sugerem o envolvimento direto entre o insulto hipóxico-isquêmico e os marcadores NF-kB e VEGFR-1, visto que se observou uma imunoexpressão diminuída destes biomarcadores nos pacientes asfixiados. Além disso, sugere-se que não houve tempo suficiente para que o VEGFR-1 fosse transcrito, traduzido e expresso na superfície da membrana plasmática. Essa temporalidade pode ser observada na relação entre a expressão de NF-kB e o tempo de vida dos indivíduos que morreram em 24 horas, o que sugere que esse fator é essencial para a produção do VEGFR-1 e, portanto, para realizar o efeito remodelador necessário para neovascularizar a região afetada.

Application of iPDMS protein microarray in screening of tumor-associated antigen autoantibodies / 生物工程学报

The rapid screening of tumor markers is a challenging task for early diagnosis of cancer. This study aims to use highly sensitive chemiluminescent protein microarray technology to efficiently screen a variety of low abundance tumor related markers. A new material, termed integrated polydimethylsiloxane modified silica gel (iPDMS), was obtained by adding a surface polymerization initiator with olefin end to the conventional polydimethylsiloxane, and fixing into the three-dimensional structure of polydimethylsiloxane by thermal crosslinking through silicon hydrogen bonding. In order to make the iPDMS material resistant to non-specific protein adsorption, a poly(OEGMA) polymer brush was synthesized by surface-initiated atom transfer radical polymerization at the active initiation site. Finally, 20 tumor-related antigens were printed into the specific areas of the microarray by high-throughput spray printing technology, and assembled into 48-well detection microtiterplates of the iPDMS microarray. It was found the VEGFR and VEGF121 autoantibodies that obtained from 8 common tumors (breast cancer, lung cancer, colon cancer, gastric cancer, liver cancer, leukemia, lymphoma and ovarian cancer) can be used as potential tumor markers. The chemiluminescence labeled iPDMS protein microarray can be used for the screening of tumor autoantibodies at early stage.

Effect of inhibitor of differentiation 1 deficiency on ocular neovascularization / 上海交通大学学报（医学版）

Objective • To study the effect of inhibitor of differentiation 1 (ID1) on ocular neovascularization. Methods • The oxygen-induced retinal neovascularization (OIR), laser-induced choroidal neovascularization (CNV) and over-expression of vascular endothelial growth factor (VEGF) (Rho-VEGF) transgenic mice were established. The localization and mRNA level of ID1 in retina of OIR mice and Rho-VEGF transgenic mice were determined by immunofluorescence staining and quantitative real-time PCR. Mice deficient in ID1 (ID1-/-) were used to induce retinal neovascularization in accordance with the above three models, and to compare the changes of ID1 on the number of retinal, subretinal and choroidal neovascularization areas. In order to explore the role ID1 in neovascularization, the numbers and areas of retinal, subretinal and choroidal neovascularization in the mice models with or without ID1 deficiency were compared. Its effect on the related factors, i.e. hypoxia-inducible factor-1α (HIF-1α), VEGF and vascular endothelial growth factor receptor 1/2 (VEGFR1/2) were also observed. Results • Mice deficient in ID1 showed a significant reduction in the area of neovascularization in these three models(P<0.05). Mice lacking ID1 showed reduced levels of HIF-1α, VEGF and VEGFR 1. Conclusion • ID1 promotes the expression of HIF-1α, VEGF and VEGFR1 in the retina and choroidal neovascularization during hypoxia and oxidative injury.

Effect of inhibitor of differentiation 1 deficiency on ocular neovascularization / 上海交通大学学报（医学版）

Objective · To study the effect of inhibitor of differentiation 1 (ID1) on ocular neovascularization. Methods · The oxygen-induced retinal neovascularization (OIR), laser-induced choroidal neovascularization (CNV) and over-expression of vascular endothelial growth factor (VEGF) (Rho-VEGF) transgenic mice were established. The localization and mRNA level of ID1 in retina of OIR mice and Rho-VEGF transgenic mice were determined by immunofluorescence staining and quantitative real-time PCR. Mice deficient in ID1 (ID1-/-) were used to induce retinal neovascularization in accordance with the above three models, and to compare the changes of ID1 on the number of retinal, subretinal and choroidal neovascularization areas. In order to explore the role ID1 in neovascularization, the numbers and areas of retinal, subretinal and choroidal neovascularization in the mice models with or without ID1 deficiency were compared. Its effect on the related factors, i.e. hypoxia-inducible factor-1α (HIF-1α), VEGF and vascular endothelial growth factor receptor 1/2 (VEGFR1/2) were also observed. Results · Mice deficient in ID1 showed a significant reduction in the area of neovascularization in these three models (P＜0.05). Mice lacking ID1 showed reduced levels of HIF-1α, VEGF and VEGFR 1. Conclusion · ID1 promotes the expression of HIF-1α, VEGF and VEGFR1 in the retina and choroidal neovascularization during hypoxia and oxidative injury.

Is the effect of melatonin on vascular endothelial growth factor receptor-2 associated with angiogenesis in the rat ovary?

OBJECTIVES Vascular endothelial growth factor (VEGF) and its receptors play important roles in angiogenesis. Melatonin plays an important role in gonadal development; thus, its effect on the reproductive system is evident. We investigated the influence of melatonin on the expression of VEGF, vascular endothelial growth factor receptor-1 (VEGFR1) and vascular endothelial growth factor receptor-2 (VEGFR2), as well as on changes in oxidative stress markers and follicle numbers in rat ovaries. METHODS For this purpose, 45 Wistar rats were separated into the following groups: Group 1, control; Group 2, vehicle; and Group 3, melatonin. Rats in Group 3 were treated with melatonin at 50 mg/kg/day for 30 days. The effects of melatonin on the expression of VEGF, VEGFR1 and VEGFR2 were established by immunohistochemistry analysis. The effects of melatonin on antioxidant enzyme activities were demonstrated by spectrophotometric analysis. RESULTS Based on immunohistochemistry analysis, VEGFR2 was predominantly localized to theca cells in the ovary. Our data indicate that melatonin treatment can significantly increase VEGF and VEGFR1 expression in the ovary ( p <0.05). Additionally, the number of degenerated follicles significantly decreased with melatonin treatment ( p <0.05). Melatonin administration also led to significant increases in antioxidant enzyme levels in the ovary. CONCLUSION Melatonin treatment exerts protective effects on follicles against increased lipid peroxidation through modulating tissue antioxidant enzyme levels. These effects may be related to angiogenesis and antioxidant activities.

The expression and significance of HIF-1α,VEGF, VEGFR-1 and VEGFR-2 in missed abortion patients / 安徽医科大学学报

Objective To study the expressions and clinical significance of hypoxia inducible factor-1α( HIF-1α) ,vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor-1 (VEGFR-1) and vascular endothelial growth factor receptor-2 (VEGFR-2) in the villi tissue of patients with missed abortion (MA) and early pregnant women. To explore the influence of hypoxia on placenta angiogenesis in missed abortion. Methods To de-tect the expression of HIF-1α,VEGF,VEGFR-1 and VEGFR-2 in the villi tissue of 35 cases of MA patients( re-search group) and 32 early pregnant women( control group) who underwent induced abortion surgery by immuno-histochemistry,and the results were statistical analysis. Results HIF-1α,VEGF,VEGFR-1 and VEGFR-2 were ex-pressed both in the villi tissue of research group and the control group. The expression of HIF-1α was higher in the research group than in the control group(P<0. 05). The expressions of VEGF,VEGFR-2 were significantly lower in the research group than in the control group ( P<0 . 05 ); the expression of HIF-1α was positively correlated with VEGF,VEGFR-1and VEGFR-2 in the control group, and the expression of HIF-1α was negatively correlated with VEGF and VEGFR-2 in the research group (P<0. 05),which had no correlation with VEGFR-1. Conclusion The expression of HIF-1α in the villi tissue of MA is up-regulated, the blood vessel formation of placenta decreases by adjusting the VEGF and VEGFR-2 genes transcription,which maybe is the important causes for MA.

Immunohistochemical expression of p53, BCL-2, BAX and VEGFR1 proteins in nephroblastomas / A expressão imuno-histoquímica das proteínas p53, BCL-2, BAX e VEGFR1 em nefroblastomas

INTRODUCTION: Nephroblastoma or Wilms' tumor is the most frequent renal cancer in children. Although its prognosis is favorable for most patients, it may relapse or have a fatal outcome. The characterization of risk groups by applying immunohistochemical biomarkers aims to adapt the treatment to its corresponding group as well as to reduce relapses and fatal outcome. p53, B-cell lymphoma 2 (BCL-2), BCL-2 associated protein X (BAX) and vascular endothelial growth factor receptor 1 (VEGFR1) are among the most widely studied biomarkers, which are related to the apoptotic pathway, DNA repair and neovascularization. OBJECTIVE: The objective of this study is to assess the immunohistochemical expression of p53, BCL-2, BAX and VEGFR1 in samples of human nephroblastoma and to correlate them with clinicopathological prognostic factors. MATERIAL AND METHODS: Twenty-nine surgical specimens of nephroblastoma diagnosed from 1994 to 2007 were selected from the Anatomopathological Service of two hospitals in Curitiba. The immunohistochemical analysis of tissue microarrays was performed through immunoperoxidase staining and the yielded results were compared with clinicopathological prognostic factors. RESULTS: The major immunohistochemical expression of VEGFR1 in blastema and epithelium presented positive association with the risk group. Hence this may be related to higher vascular neoplastic invasion apparently caused by the endothelial growth factor, which maximizes the chances of metastasis and ultimately changes tumor staging, risk group and clinical evolution. CONCLUSIONS: The immunohistochemical expression of VEGFR1 substantiated a directly proportional association with the nephroblastoma risk group.

INTRODUÇÃO: O nefroblastoma, ou tumor de Wilms, é a neoplasia renal mais frequente na infância. Embora o prognóstico seja favorável para a maioria dos pacientes, muitos evoluem para recidiva ou óbito. A caracterização de grupos de risco por meio de marcadores imuno-histoquímicos tem por objetivo adequar o tratamento ao grupo de risco e reduzir recidivas e óbitos. Entre os marcadores mais estudados estão p53, B-cell lymphoma 2 (BCL-2), BCL-2 associated protein X (BAX) e fator de crescimento vascular endotelial e seu receptor 1 (VEGFR1), relacionados com a via apoptótica, o reparo do ácido desoxirribonucleico (DNA) e a neovascularização. OBJETIVO: O objetivo deste estudo é avaliar a expressão imuno-histoquímica de p53, BCL-2, BAX e VEGFR1 em amostras de nefroblastoma humano e correlacioná-las com fatores prognósticos clínico-patológicos. MATERIAL E MÉTODO: Foram utilizadas 29 amostras de nefroblastomas retiradas dos Serviços de Anatomia Patológica de dois hospitais de Curitiba, com diagnósticos entre 1994 e 2007. Por meio da técnica de imunoperoxidase, com as amostras em arranjo tecidual em matriz, realizou-se análise da imuno-histoquímica desses marcadores e sua comparação com fatores prognósticos clínico-patológicos. RESULTADOS: A maior imunoexpressão de VEGFR1 nos componentes blastematoso e epitelial mostrou associação positiva ao grupo de risco, sendo que isso poderia estar relacionado com a maior capacidade de invasão vascular neoplásica que pode ser conferida por esse fator de crescimento endotelial, aumentando, assim, as chances de metástases e alterando o estadimento, o grupo de risco e a evolução clínica. CONCLUSÃO: A expressão imuno-histoquímica aumentada de VEGFR1 mostrou associação diretamente proporcional ao grupo de risco dos pacientes com nefroblastoma.

Expression of Aquaporin-1, Vascular Endothelial Growth Factor and Vascular Endothelial Growth Factor Receptor-1 in the Peritoneal Tissues of Peritoneal Dialysis Patients / 대한신장학회지

PURPOSE: AQP-1 (Aquaporin-1) and VEGF (vascular endothelial growth factor) are known to play an important role in ultrafiltration in peritoneal dialysis. The aim of this study was to evaluate the expression of AQP-1 and VEGF and VEGFR-1 (VEGF type 1 receptor) in peritoneums obtained from uremic non-dialyzed patients and peritoneal dialysis patients and to see if expression of these molecules are correlated with each other and with pathological findings in peritoneum. METHODS: Peritoneal expressions of AQP-1, VEGF and VEGFR-1 were examined by immunohistochemistry using specific antibody to each molecule. The degree of vascular proliferation and inflammation in peritoneal tissues were assessed semi-quantitatively by a single pathologist. RESULTS: AQP-1, VEGF and VEGFR-1 were mainly expressed in the vascular endothelial cells in the peritoneum. No significant difference in peritoneal expression of these molecules was found according to the clinical situations in which peritoneal tissues were obtained. The degree of expression of AQP-1 and VEGF were related to each other but not related to expression of VEGFR-1. The expressions of AQP-1 and VEGF were related to the vascular proliferation. The expression of AQP-1 was also related to inflammation. CONCLUSION: In end-stage renal disease patients before and after initiation of peritoneal dialysis, the peritoneal expressions of AQP-1 and VEGF were related to vascular proliferation. Inflammation might have some influence in expression of AQP-1.

Angiogenic growth factor responses to long-term treadmill exercise in mice.

We sought to determine whether VEGF and other angiogenic growth factors and their receptors might be subject to negative feedback regulation during two weeks of treadmill-exercise conditioning in inbred strains of mice. C57BL/6 mice exhibited greater VEGF mRNA and protein responses in gastrocnemius muscle to a single bout of treadmill exercise compared to BALB/c mice. The patterns of VEGF, VEGFR1, VEGFR2, Ang2 and Tie2 mRNA expression in gastrocnemius muscles of C57BL/6 mice during long-term exercise support the hypothesis that they may be subject to negative feedback regulation. The combination of expression patterns for growth factors and their receptors suggests that multiple layers of control mechanisms may exist to prevent angiogenesis following a single bout of exercise and to promote angiogenesis following long-term exercise.

Free vascular endothelial growth factor and its soluble receptor in peritoneal fluid from endometriosis patients / 第三军医大学学报

Objective To better understand the regulation of the soluble receptor of vascular endothelial growth factor (VEGF), s-VEGFR-1 (or soluble fms-like tyrosine kinase, SFlt-1), in angiogenetic process in endometriosis. Methods Levels of free VEGF and s-VEGFR-1 were measured by enzyme-linked immunosorbent assay (ELISA) in peritoneal fluid from 28 subjects with surgically confirmed endometriosis, and 10 controls with no clinical evidence of the disease and other diseases. Meanwhile, we calculated a VEGF activity index by means of the ratio VEGF/s-VEGFR-1. Results We found higher VEGF concentration in endometriotic lesions than controls (P0.05). VEGF activity index in controls, stage Ⅰ-Ⅱ and stages Ⅲ-Ⅳ of endometriosis was 0.310, 0.276 and 0.273, respectively. VEGF and s-VEGFR-1 concentration were also higher in proliferative phase than in secretory phase in endometriotic lesions (P