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SUMMARY: Neuropeptide calcitonin gene-related peptide (CGRP) is a neurotransmitter related to vasculogenesis during organ development. The vascular endothelial growth factor A (VEGF-A) is also required for vascular patterning during lung morphogenesis. CGRP is primarily found in organs and initially appears in pulmonary neuroendocrine cells during the early embryonic stage of lung development. However, the relationship between CGRP and VEGF-A during lung formation remains unclear. This study investigates CGRP and VEGF-A mRNA expressions in the embryonic, pseudoglandular, canalicular, saccular, and alveolar stages of lung development from embryonic day 12.5 (E12.5) to postnatal day 5 (P5) through quantitative real-time polymerase chain reaction (qRT-PCR) and in situ hybridization. Further, we analyzed the expression of CGRP via immunohistochemistry. The VEGF-A mRNA was mainly scattered across the whole lung body from E12.5. CGRP was found to be expressed in a few epithelial cells of the canalicular and the respiratory bronchiole of the lung from E12.5 to P5. An antisense probe for CGRP mRNA was strongly detected in the lung from E14.5 to E17.5. Endogenous CGRP may regulate the development of the embryonic alveoli from E14.5 to E17.5 in a temporal manner.
El péptido relacionado con el gen de la calcitonina (CGRP) es un neurotransmisor vinculado con la vasculogénesis durante el desarrollo de órganos. El factor de crecimiento endotelial vascular A (VEGF-A) también se requiere para el patrón vascular durante la morfogénesis pulmonar. El CGRP se encuentra principalmente en los órganos y aparece inicialmente en las células neuroendocrinas pulmonares durante la etapa embrionaria temprana del desarrollo pulmonar. Sin embargo, la relación entre CGRP y VEGF-A durante la formación de los pulmones sigue sin estar clara. Este estudio investiga las expresiones de ARNm de CGRP y VEGF-A en las etapas embrionaria, pseudoglandular, canalicular, sacular y alveolar del desarrollo pulmonar desde el día embrionario 12,5 (E12,5) hasta el día postnatal 5 (P5) a través de la reacción en cadena de la polimerasa cuantitativa en tiempo real. (qRT-PCR) e hibridación in situ. Además, analizamos la expresión de CGRP mediante inmunohistoquímica. El ARNm de VEGF-A se dispersó principalmente por todo parénquima pulmonar desde E12,5. Se encontró que CGRP se expresaba en unas pocas células epiteliales de los bronquiolos canaliculares y respiratorios del pulmón desde E12,5 a P5. Se detectó fuertemente una sonda antisentido para ARNm de CGRP en el pulmón de E14,5 a E17,5. El CGRP endógeno puede regular el desarrollo de los alvéolos embrionarios de E14,5 a E17,5 de manera temporal.
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Animals , Mice , Calcitonin Gene-Related Peptide/metabolism , Vascular Endothelial Growth Factor A/metabolism , Lung/growth & development , Lung/embryology , Immunohistochemistry , In Situ Hybridization , Neurotransmitter Agents , Neovascularization, PhysiologicABSTRACT
BACKGROUND: The distinct arterial and venous cell fates are dictated by a combination of various genetic factors which form diverse types of blood vessels such as arteries, veins, and capillaries. We report here that YULINK protein is involved in vasculogenesis, especially venous formation. METHODS: In this manuscript, we employed gene knockdown, yeast two-hybrid, FLIM-FRET, immunoprecipitation, and various imaging technologies to investigate the role of YULINK gene in zebrafish and human umbilical vein endothelial cells (HUVECs). RESULTS: Knockdown of YULINK during the arterial-venous developmental stage of zebrafish embryos led to the defective venous formation and abnormal vascular plexus formation. Knockdown of YULINK in HUVECs impaired their ability to undergo cell migration and differentiation into a capillary-like tube formation. In addition, the phosphorylated EPHB4 was decreased in YULINK knockdown HUVECs. Yeast two-hybrid, FLIM-FRET, immunoprecipitation, as well as imaging technologies showed that YULINK colocalized with endosome related proteins (EPS15, RAB33B or TICAM2) and markers (Clathrin and RHOB). VEGF-induced VEGFR2 internalization was also compromised in YULINK knockdown HUVECs, demonstrating to the involvement of YULINK. CONCLUSION: This study suggests that YULINK regulates vasculogenesis, possibly through endocytosis in zebrafish and HUVECs. Key points Knockdown of YULINK with morpholino in embryos of double transgenic zebrafish exhibited abnormal venous formation. Tube formation and phosphorylated EPHB4 were decreased in YULINK knockdown HUVECs. FLIM-FRET, immunoprecipitation, as well as other imaging technologies showed that YULINK colocalized with endosome related proteins (EPS15, RAB33B and TICAM2) and endosome markers (Clathrin and RHOB). Knockdown of YULINK decreased the internalization of VEGF and VEGFR2 in HUVECs.
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Humans , Animals , Saccharomyces cerevisiae , Zebrafish/genetics , Cell Differentiation , Cell Movement , Neovascularization, Physiologic , Human Umbilical Vein Endothelial CellsABSTRACT
@#With the development of biomimetic technology, more and more in vitro models are used to simulate human physiological and pathological processes.These in vitro models can solve some scientific problems, such as studying drug effects in real-timely and visually.As an in vitro model, organ-on-a-chip provides novel means and methods for basic and applied science.The vascularized organ-on-a-chip, as a special kind of organ-on-a-chip, can better simulate the structure and function of human blood vessels.In this review, we summarized the structure and function of different vascularized organ-on-a-chip, analyzed the application of vascularized organ-on-a-chip in simulating physiological and pathological processes, and discussed the advantages and problems to be solved of vascularized organ-on-a-chip as a new in vitro model.Finally, the application of vascularized organ-on-a-chip is proposed.
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Aim To investigate the effect of Exendin-4 high glucose and the function of silent information reg- on endothelial progenitor cells ( EPCs) induced by ulator 1 (SIRT1 ).Methods EPCs were isolated and cultured by density gradient centrifugation from peripheral blood of healthy volunteers.Different concentrations of Exendin-4( 12.5, 50, 100, 200 (xmol • L"1) induced EPCs were respectively performed by activity test to select the appropriate concentration.The optimal concentrations of Exendin-4 and high glucose (25 mmol • L 1) were incubated together for 72 h to detect the functional activities of EPCs.The capabilities of migration, adhension and tube formation of EPCs in vitro were detected respectively.The levels of LDH and MDA in EPCs were detected.The mRNA expressions of TNF-a, 1L-1 p and IL-6 in EPCs were measured by real-time fluorescent quantitative PCR ( RT-qPCR ).The protein expression of SIRT1 , P53 and Ac-P53 in EPCs were determined by Western blot.Results Ex- endin-4 could increase the viability of EPCs induced by j J high glucose in a dose-dependent manner, especially for 50 (xmol • L "1 (P <0.05 ).Hie results of Western blot showed that the protein expression of SIRT1 was significantly enhanced by 50 |xmol • L"' Exendin-4 treatment ( P < 0.05 ).Compared with high glucose group, Exenclin-4 significantly increased the migration, adhesion, tube formation of EPCs (P<0.05) and decreased the level of LDH and MDA ( P < 0.05 ).The mRNA expression of TNF-cx, lL-(3 and IL-6 in EPCs also decreased (P < 0.05).However, the protective effects of Exendin-4 could he significantly blocked by SIRT1 inhibitor ( EX-527) (P<0.05).In addition, the S1HT1 agonist ( SHT1720 ) could also improve the dysfunction of EPCs induced by high glucose ( P < 0.05).Conclusions Exendin-4 can improve the viability of human EPCs, restore the EPCs normal function, reduce high glucose-induced oxidative damage, and reduce the releases of inflammatory cvtokines un- j j der high glucose condition, which may be related to the regulation of S1HT1/P53 signaling pathway.
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Aim To investigate the effect of Exendin4 on proliferation, migration, adhesion and senescence of endothelial progenitor cells (EPCs) in type I diabetic mice and its possible mechanism. Methods EPCs from 6-month diabetic mice were isolated and cultured by density gradient centrifugation. Cells were treated with different concentrations of Exendin-4 (1, 5, 10, 25 μmol · L
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BACKGROUND: Fracture is a common orthopedic disease in clinic. Although most fractures can heal by primary bone healing through surgical treatments, such as rigid internal fixation, malunion is liable to occur in some circumstances, eventually leading to bone defects. Neovascularization at the bone defect site plays a vital role in bone healing. Based on the pro-angiogenic ability of endothelial progenitor cells (EPCs), its capacity for bone regeneration and repair has gradually become the focus of attention. OBJECTIVE: To review the use of EPCs transplantation to promote vasculogenesis and osteogenesis in bone defects. METHODS: With the key words of “endothelial progenitor cells (EPCs), angiogenesis, vasculogenesis, therapy/treatment, bone defect” in Chinese and English, we performed a computer-based search in CNKI, WanFang and PubMed databases for relevant articles published from 1986 to 2019. Finally, 58 eligible articles were included in result analysis. During the literature retrieval, we followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses. RESULTS AND CONCLUSION: Bone defects can promote the activation and homing of EPCs. EPCs can promote vasculogenesis and further trigger bone regeneration. There are still some problems to be solved before the application of EPCs in the treatment of bone defects. More experiments need to be carried out to investigate the exact mechanism of EPCs in vasculogenesis. More clinical trials are warranted, providing data support for future clinical applications and giving better treatment to patients disabled by bone defects.
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@#Objective: To investigate the inhibitory effect of Triptolide on vasculogenesis of human cervical microvascular endothelial cells, and to explore the mechanism. Methods: Human cervical microvascular endothelial cells (HCerMECs) were used as research subject, and treated with different concentrations of Triptolide (0, 5, 10, 20 and 40 ng/ml) i n v i t r o . The effect of Triptolide on cell proliferation was determined by CCK-8, the cell migration ability was detected by Transwell assay while the expression of vascular endothelial growth factor (VEGF) was examined by western blotting. Results: Triptolide inhibited the proliferation and migration of HCerMECs in a dose-dependent manner ( P <0.05). In addition, Triptolide could inhibit the expression of VEGF in HCerMECs in a concentration-dependent manner. Conclusions: Triptolide could inhibit the proliferation and migration activity of HCerMECs which is related with the suppression of VEGF expression.
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Objective: To investigate effect of Yanghe Huayan Tang and phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 in intervening expressions of human epidermal growth factor receptor human epidermal growth factor receptor-2(HER-2), PI3K and phosphorylated serine/threonine kinase (p-Akt)in PI3K/Akt pathway of breast cancer cell line SK-BR-3 (HER-2 high expression) in vitro, and study intervention mechanism of Yanghe Huayan Tang. Method: Yanghe Huayan Tang intestinal absorption fluid was prepared, breast cancer cell strain SK-BR-3 was divided into blank group, Yanghe Huayan Tang group, LY294002 group, LY294002+Yanghe Huayan Tang group. The concentration was respectively 125 g·L-1 for Yanghe Huayan Tang, 0.05 g·L-1 for LY294002, 125 g·L-1 for Yanghe Huayan Tang+LY294002.The expressions of HER-2, PI3K and p-Akt protein were detected by immunohistochemistry and Western blot after 24 h. The most appropriate and effective concentration was obtained through extensive experiments. The expressions of HER-2, PI3K and p-Akt were detected by reverse transcription polymerase chain reaction (RT-PCR). Result: Compared with blank group, LY294002 group, and LY294002+Yanghe Huayan Tang group could inhibit protein and mRNA expressions of HER-2, PI3K, p-Akt in breast cancer cell lines(PPPPConclusion: Yanghe Huayan Tang can inhibit angiogenesis and invasion of breast cancer. Its main mechanism is expressed by intervening PI3K/Akt pathway, reducing expressions of HER-2, PI3K and p-Akt in pathway.
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@#Objective To investigate the role of kinase insert domain containing receptor (KDR) positive cells in the formation of cardiospheric structure, myocardium and vessels. Methods Twenty-four Wistar rats weighting 250 g were selected. Cardiosphere-derived cells were isolated by enzymatic digestion of rat hearts, and their immunological phenotypes were analyzed by using fluorescence-activated cell sorting (FACS). The cardiomyogenic and vasculogenic potential was diagnosed by immunohistochemistry. Results KDR positive cells grew exponentially and formed cell clusters. It also could generate myocardial precursor cells (cardiac troponin T positive). And these cells can develop spontaneous contraction activity in vitro. Meanwhile, KDR positive cells formed many vessel-like structures through a budding process. Conclusion KDR positive cells form cardiospheric structure in vitro culture, and exhibit differentiation potential towards the cardiac and vascular cells. Therefore, KDR positive cells may have a broad prospect of clinical application as cell donors.
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La preeclampsia constituye una enfermedad de origen obstétrico que implica un aumento de la morbilidad y la mortalidad materna y perinatal. A tales efectos se llevó a cabo una extensa revisión bibliográfica con el fin de exponer las teorías más actualizadas en relación con la fisiopatología de la preeclampsia, que será un material de consulta valioso para los obstetras, quienes profundizarán en la génesis de la enfermedad, lo cual repercute en la toma de mejores decisiones.
Preeclampsia constitutes a disease of obstetric origin that implies an increase of the maternal and perinatal morbidity and mortality. To such effects an extensive literature review was carried out with the purpose of exposing the most updated theories in relation to the pathophysiology of preeclampsia that will be a valuable material for the obstetricians who will deepen in the genesis of the disease, which rebounds in a better decision making.
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Pre-Eclampsia/physiopathology , Eclampsia , Genomics , Proteomics , Neovascularization, PathologicABSTRACT
PURPOSE: The aim of this study was to investigate whether plasma angiopoietin-1 (Ang-1) and angiopoietin-2 (Ang-2) concentrations in pregnant women with chronic hypertension are different from those of normotensive pregnant women. METHODS: This hospital-based case-control study consisted of 35 pregnant women with chronic hypertension who delivered at Seoul National University Hospital. Normotensive pregnant women (n=70) were selected as controls, matched with maternal age, gestational age at delivery and birthweight. Maternal blood was drawn at the time of admission for delivery and plasma was separated and stored. The plasma Ang-1 and Ang-2 levels were measured by ELISA. Statistical analysis was done with Mann-Whitney U test, Fisher's exact test and Spearman rank correlation test using SPSS. RESULTS: Median (range) maternal age, gestational age and birthweight were 33 years (24-42), 38 weeks (32-41), and 3.08 kg (1.13-4.01). Pregnant women with chronic hypertension had significantly higher median Ang-1 and Ang-2 levels than normotensive pregnant women (for Ang-1 : median 4,111 pg/mL, range 1,415-30,172 vs. median 2,824 pg/mL range 662-14,512, P=0.015, for Ang-2 : median 5,637 pg/mL, range 1,131-29,327 vs. median 3,345 pg/mL, range 609-24,467, P=0.039). CONCLUSION: Maternal plasma Ang-1 and Ang-2 levels were elevated in pregnant women with chronic hypertension compared with normotensive pregnant women. Further study is needed to determine if this change is a cause or a compensatory mechanism to chronic hypertension.
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Female , Humans , Angiopoietin-1 , Angiopoietin-2 , Case-Control Studies , Enzyme-Linked Immunosorbent Assay , Gestational Age , Hypertension , Maternal Age , Plasma , Pregnant Women , SeoulABSTRACT
In order to test if nestin is a useful marker for various types of progenitor cells, we explored nestin expression in the retina during development. Nestin expression was co-evaluated with bromodeoxyuridine (BrdU) labeling and Griffonia simplicifolia isolectin B4 (GSIB4) histochemistry. Nestin immunoreactivity appears in cell soma of dividing neural progenitor cells and their leading processes in retinas from embryonic day (E) 13 to E20, in accordance with a BrdU-labeled pattern. At postnatal day (P) 5, it is restricted to the end feet of Muller cells. BrdU-labeled nuclei were mainly in the inner part of the inner nuclear layer in postnatal neonates. The retinal vessels demarcated with GSIB4-positive endothelial cells were first distributed in the nerve fiber layer from P3. Afterward the vascular branches sprouted and penetrated deeply into the retina. The endothelial cells positive for GSIB4 and the pericytes in the microvessels were additionally immunoreactive for nestin. Interestingly, the presumed migrating microglial cells showing only GSIB4 reactivity preceded the microvessels throughout the neuroblast layer during vascular sprouting and extension. These findings may suggest that nestin expression represents the proliferation and movement potential of the neural progenitor cells as well as the progenitor cells of the endothelial cell and the pericyte during retinal development. Thus, Muller glial cells might be potential neural progenitor cells of the retina, and the retinal microvasculature established by both the endothelial and the pericyte progenitor cells via vasculogenesis along microglia migrating routes sustains its angiogenic potential.
Subject(s)
Humans , Infant, Newborn , Bromodeoxyuridine , Carisoprodol , Endothelial Cells , Foot , Griffonia , Intermediate Filament Proteins , Lectins , Microglia , Microvessels , Nerve Fibers , Nerve Tissue Proteins , Neurogenesis , Neuroglia , Pericytes , Plant Lectins , Retina , Retinal Vessels , Retinaldehyde , Stem CellsABSTRACT
Objective To study the effect of granulocyte-macrophage colony-stimulating factor(GM-CSF)on angiogenesis of rat with acute myocardial injury induced by isoproterenol(Iso). Methods A total of 60 adult male Wistar rats were randomly divided into 3 groups: normal control group, GM-CSF pretreatment group (GM-CSF group), and lso injury group, 20 rats in each group. GM-CSF group was administered recombinant human(rh)GM-CSF(5.0 μg/kg), through tail intravenous injection once a day for three days. Then the GM-CSF group and the Iso injury group were anesthetized by intraperitoneal injection of lso( 15.0 mg/kg) once a day for three days. The same dose of saline was administered in the same way to the control rats. Ten days after injection, pathological changes of myocardial damage and infarct area were examined by immunohistochemistry. The mRNA expression levels of polypeptide antigen (CD34), vascular endothelial growth factor (VEGF) and its receptor KDR/flk- 1 were measured by RT-PCR. Results The difference of myocardial necrosis area between groups was statistically significant(F=10.07, P < 0.01), in which GM-CSF group[(37.37 ± 12.98)%] was significantly less than Iso injury group[(45.51 ±14.96)%, P < 0.05]. The difference of myocardial neovascularization density index of rats between groups was statistically significant ( F = 25.54, P < 0.05 ), in which GM-CSF group [(3980.05 ± 477.22) No/mm2] was significantly higher than Iso injury group((2605.93±361.49)No/mm2,P<0.01).The differences of myocardial CD34,VEGF,KDR/flk-1 mRNA expression between groups were statistically significant(F=17.83,4.29,4.10,all P<0.01).Compared to Iso mjury group[CD34(23.85±6.06),VEGF(31.80±8.05),KDR/flk-1(30.16±8.01)]were higher in the GM-CSF group[CD34(44.04±10.13),VEGF(49A±11.59),and KDR/flk-1(46A9±7.90),all P<0.01].The expressions of myocardiM VEGF mRNA and its receptor KDR/flk-1 mRNA was positively correlated(r=0.725,R2=0.526,P<0.01).Conclusions GM-CSF prelreatmcnt increases the density ofnew blood vessels in myocardium,and reduces the Iso-induced myocardial injury in rats.
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The formation of the coronary vasculature is a fundamental event in heart development and involves a series of carefully regulated temporal events that include vasculogenesis and angiogenesis. This review focuses the knowledge concerning the formation of the coronary arteries available so far and some molecular mechanisms involved in this process. Understanding coronary embryogenesis is important for interventions regarding adult cardiovascular diseases as well as those necessary to correct heart congenital defects. The insight of the coronary artery development as a result of ingrowth changed the understanding of several congenital coronary artery variations and anomalies described in gross anatomy.
La formación de la vascularización coronaria es un acontecimiento fundamental en el desarrollo del corazón e implica una serie de eventos temporales cuidadosamente regulados que incluyen vasculogénesis y angiogénesis. Esta revisión se focaliza en el conocimiento sobre la formación de las arterias coronarias y algunos mecanismos moleculares implicados en este proceso. Entender la embriogénesis coronaria es importante para las intervenciones relacionadas con las enfermedades cardiovasculares en los adultos, así como también, para corregir defectos congénitos del corazón. La idea del desarrollo de la arteria coronaria, como resultado del crecimiento interno, cambió la comprensión de diversas variaciones y anomalías congénitas de estas arterias descritas en la anatomía macroscópica.
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Humans , Male , Female , Coronary Vessels/anatomy & histology , Coronary Vessels/embryology , Coronary Vessels/ultrastructure , Morphogenesis/genetics , Neovascularization, Physiologic/physiologyABSTRACT
The vaseularization of tissue-engineered bone is the key problem which the development and employment of large sized tissue-engineered bone.The vascular endothelial cell has a great effect on promoting vascularization in tissue-engineered bone.Vascularizations fall into two modes of vaseulogenesis and angiogenesis according to differences in source of endothelial cells.Co-culture of osteoblasts and vascular endothelial cells has better result than single culture of each kind of cells.Different ways of improving the vascularization,such as searching for new source of vascular endothelial cell,co-culture and in vivo experiment are investigated to meet the challenge of bone tissue engineering.
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Tipo de Estudio: retrospectivo, descriptivo. Objetivo: identificar los factores de riesgo y su influencia en la retinopatía del prematuro (ROP). Método: Se tomaron 74 recién nacidos pretérmino en el hospital Gineco-Obstétrico Enrique C. Sotomayor con factores de riesgo conocidos y se les realizó el examen de fondo de ojo mediante oftalmoscopía indirecta entre la cuarta y octava semanas de vida extrauterina; y sus respectivos controles hasta el alta oftalmológica. Resultados: de la población estudiada, 48 pacientes completaron los controles sin presentar la patología, y los 25 pacientes restantes (34) presentaron ROP en distintos grados. Tomando en cuenta los factores de riesgo estudiados y su repercusión sobre la patología: -La edad gestacional promedio de neonatos afectos fue de 32.8 semanas. -Tiempo promedio de exposición a oxigenoterapia de 10 días. -Promedio de peso al nacer 1325.5 gramos. Conclusiones: en este estudio la inmadurez neonatal fue el factor más determinante en la incidencia de esta patología, seguido del bajo peso al nacer y del tiempo de exposición a oxigenoterapia; y dado el gran numero de nacimientos pretérmino en el mundo y mayor aun en países en desarrollo como el nuestro, es menester hacer hincapié en la necesidad de una correcta implementación y organización interhospitalaria del programa de detección temprana de Retinopatía del Prematuro en nuestra ciudad, y de esta manera evitar el inicio de una vida en penumbra.
Type of study: Retrospective, descriptive. Objective: Identify the risk factors and its influence in retinopathy in premature patients. Method: A total of 74 preterm newborn at the Enrique C. Sotomayor Gineo-Obstetric Hospital with known risk factors were included in the study. They underwent an indirect ophtalmoscopy between the fourth and eight week of life and were monitored. Results: From the total population, 48 patients did not have symptoms of the pathology while being monitored and 25 patients left (34) had different stages of retinopathy. Our findings were: The average gestational age of the neonates was 32.8 weeks, the average time of oxygen therapy was 10 days and the average weight was 1325.5 grams. Conclusions: In this study prematurity was the factor that determined the incidence of the pathology, followed by low birth weight and the period received oxygen therapy. Due to the high incidence of premature births in the world and even higher in third world countries like ours it is important to organize a program for the early detection of retinopathy in premature newborns.
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Male , Female , Infant, Newborn , Retinopathy of Prematurity , Risk Factors , Infant, Low Birth Weight , Infant, Premature , Infant, Very Low Birth Weight , Refractive Errors , Retinal DetachmentABSTRACT
Objective To study some biological properties of mouse bone marrow stromal cells(MSCs)and their ability to differentiate endothelial cells and surviving ability in ischemic tissue,and provide an experimental foundation for applying MSCs to ischemic repair.Methods After the tibias and femurs were dissected from 5-to 6-week-old mice from Jan.2005 to Nov.2005,the marrow was flushed out with ice-cold DMEM/F12 medium.The mononuclear cells of the marrow were obtained with density gradient centrifugation and the plated and cultured in DMEM/F12 medium.The cultured cells in vitro were induced with endothelial cell growth supplement.The ability of MSCs was also was examined in ischemic tissue.Results The adherent fibroblast-shaped cells approached confluence in single layer 12~16 d after plating.The cultured MSCs in vitro differentiated into endothelium.Numerous scattered DAPI-labeled cells were found in the specimen seven days after implantation,and hematoxylin and eosin staining of the adjacent section showed concordance between dense hematoxylin staining and presence of DAPI epifluorescence,and there was no obvious inflammatory response.Conclusion The subcultured MSCs possess potential to differentiate into endothelial cells.MSCs show stable growth in vitro,easy survival in the subculture and rapid proliferation in present culture condition.MSCs may be used in therapy for myocardium ischemia.
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Zishen Tiaochong therapy can improve the function of corpus luteum.The function of corpus luteum has close correlation with vasculogenesis in which the bone marrow-derived endothelial progenitor cells(EPCs)were involved.This implies the Zishen Tiaochong therapy may be correlative to vasculogenesis in corpus luteum via mobilizing bone marrow-derived EPCs.In this paper,based on the TCM theory of 'kidney(Shen) storing essence,producing bone marrow,controlling reproduction',we review the topic of Zishen Tiaochong therapy perfects the function of corpus luteum combined the new mechanisms of research advances in vasculogenesis.The possible mechanisms of promoting vasculogenesis in corpus luteum by Zishen Tiaochong therapy were discussed and analyzed.
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In the last 10 years,an increasing interest has been devoted to the study of endothelial progenitor cells(EPCs).Alteration and/or reduction of the circulating EPCs have/has been associated with different manifestation of cardiovascular disorders.Thererfore,EPCs have been extensively studied as biomarkers of cardiovascular disease and as a cell-based therapy for repair of damaged blood vessels.In the review,we focus on two methodologies used to identify and enumerate EPCs in the majority of studies up to date.
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BACKGROUND AND OBJECTIVES: Ex vivo expansion of endothelial cells is important when applying cell therapy to therapeutic angiogenesis in ischemic tissues. Endothelial precursor cells (EPCs) from the umbilical cord blood are one of adult stem cell. In order to establish the culture system for EPCs, we examined the effects of the media and matrix on the differentiation of a subset of mononuclear cells to endothelial cells, and analyzed their endothelial-lineage phenotype. MATERIALS AND METHODS: Mononuclear cells isolated from human umbilical cord blood were cultured in a chamber slide coated with fibronectin or gelatin in a M199 medium supplemented with 10% fetal bovine serum (FBS) (the normal medium) or with 20% FBS and ECGS (the rich medium). Changes in the morphology and the attainment of DiI-ac-LDL uptake ability were examined during a 7 day period. The attached cells were immunostained for CD31, KDR, and vWF. RESULTS: The fibronectin matrix gave rise to more attached cells than the gelatin matrix (about 1.5 fold). The numbers of attached cells were no different between the normal medium and the rich medium at day 3 and 7, and were about 12% of the seeded mononuclear cells. However, the cell size and the numbers of longer spindle-shaped cells increased with the rich medium. Moreover, there was no increase in cellular population, but a 2-3 fold increase in the cellular size between day 3 and 7. About 20-40% of the attached cells acquired the DiI-ac-LDL uptake ability at day 3, whereas more than 85% of the attached cells could be stained with fluorescent DiI-ac-LDL at day 7 (p<0.001). The attached cells after being cultured for 7 days were stained moderately with the antibodies of CD31, or KDR. However, the cells at day 7 were only weakly immunostained with the vWF antibody, whereas more than 90% of cells were strongly stained at day 14. CONCLUSION: These results suggest that a subset of mononuclear cells derived from cord blood cells can give rise to cells with an endothelial cell-like phenotype, in vitro, at high percentages, which could be applied to in vivo vasculogenesis.