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Aims: Allogeneic bone marrow (BM) has been shown to support human islet survival and function in long-term culture by initiating human islet vascularization and β-cell regeneration. Various BM subpopulations may play different roles in human islet functions and survival. In this paper we investigated the effects of BM and its subpopulations, endothelial progenitor cells (E) and mesenchymal (M) cells on human islet’s β-cell function and regeneration. Study Design: Isolation and identification of subpopulations from human bone marrow and culture with allogeneic human islet to investigate effects of different cell population on human islet function and regeneration. Place and Duration of Study: Department of Medicine, Center for Stem Cell & Diabetes Research, RWMC, Providence, RI, USA, between 2010 - 2014. Methodology: Human islets were distributed from Integrated Islet Distribution Program (IIDP) and human bone marrow (BM) was harvested by Bone marrow transplantation center at Roger Williams Hospital. BM subpopulation was identified cell surface markers through Fluorescenceactivated cell sorting, applied in flow cytometry (FACS), islet function was evaluated by human ELISA kit and β cell regeneration was evaluated by three methods of Cre-Loxp cell tracing, β cell sorting and RT-PCR for gene expression. Results: Four different BM and seven different islet donates contributed human tissues. We observed islet β-cell having self regeneration capability in short term culture (3~5 days) using a Cre-Loxp cell tracing. BM and its subtype E, M have similar benefits on β cell function during coculture with human islet comparison to islet only. However, only whole BM enables to sustain the capability of islet β-cell self regeneration resulting in increasing β cell population while single E and M individual do not significantly affect on that. Mechanism approach to explore β-cell self regeneration by evaluating transcription factor expressions, we found that BM significantly increases the activations of β-cell regeneration relative transcription factors, the LIM homeodomain protein (Isl1), homologue to zebrafish somite MAF1 (MAFa), the NK-homeodomain factor 6.1 (NKX6.1), the paired box family factors 6 (PAX6), insulin promoter factor 1 (IPF1) and kinesin family member 4A (KIF4a). Conclusion: These results suggest that BM and its derived M and E cells enable to support human islet β-cell function. However, only BM can sustain the capability of β-cell self regeneration through initiating β-cell transcriptional factors but not individual E and M cells suggesting pure E and M cells less supportive for islet long-term survival in vitro.
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Objective To investigate the effects and the possible mechanism of insulin like growth factor-1 ( IGF1 ) on proliferation and migration of human umbilical vein endothelial cells ( HUVEC) in high glucose. Methods Cultured HUVEC were divided into 5 groups( control group, high glucose group, hypertonic group, high glucose+IGF 1 group, high glucose+IGF1+ AZD5363 group) . The proliferation was detected by methyl thiazolyl tetrazolium ( MTT) , the migration was detected by transwellchamber. The mRNA expressions of serine/threonine kinase( Akt) and FoxO1 were detected by real-time PCR. The protein expressions of FoxO1/p-FoxO1 and Akt/p-Akt were detec-ted by immunohistochemisty. Results Compared with the control group, the rate of cell survival in high glucose group decreased significantly(P<0. 05), the transitional was decreased significantly(P<0. 05), the mRNA ex-pression of FoxO1 was increased significantly( P<0. 05 ) , the protein expressions of FoxO1/p-FoxO1 and Akt/p-Akt were increased. Compared with high glucose group, the cell survival rate and migration rate in high glucose+IGF1 group were increased significantly, the mRNA expression of FoxO1 was decreased significantly(P<0. 05), the protein expressions of FoxO1/p-FoxO1 and Akt/p-Akt were decreased. Compared with high glucose group, the cell survival rate and migration rate in high glucose+IGF1+AZD5363 group had no significant difference, the mR-NA expression of FoxO1 was decreased significantly ( P<0. 05 ) , the protein expressions of FoxO1/p-FoxO1 had no significant difference. The mRNA expression of Akt in five groups had no significant difference. Conclusion IGF1 can improve the proliferation and migration of endothelial cell in high glucose. It shows that IGF1 acts through Akt to promote FoxO1 phosphorylation.
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OBJECTIVE: To investigate the effects of atorvastin on function of glomerular endothelia cells in rats with chronic renal failure (CRF). METHODS: Twenty-eight male SD rats were randomly divided into 4 groups: sham operation group (control group), CRF group (model group), 8 mg · kg-1 · d-1 atorvastin treatment group (low-dosage group) and 16 mg · kg-1 · d-1 atorvastin treatment group (high-dosage group). The model of chronic renal failure was established by a two stage 5/6 nephrectomy procedure. The atorvastin treatment groups were given atorvastin by intragastric administration, and the other 2 groups were given sodium chloride. Serum creatinine (Scr), blood urea nitrogen (BUN), urine protein, total cholesterol (TCHO), triglycerides (TG), low density liporotein (LDL), AST, ALT and creatine kinase (CK) were measured after 8 weeks. The renal morphologic changes were e-valuated on periodic acid-schiff (PAS) stained sections. The CD34 and CD31 expressions in glomerulus were detected by immunohisto-chemistry method. The mRNA of ET1, eNOS and VEGF were detected by RT-PCR. RESULTS: The Scr, BUN and urine protein levels in atorvastin treatment groups were significantly lower than those in CRF model group (P < 0.05). Reanl pathological injuries were improved by atorvastin treatment in dose-dependent manner. There were no significant differences in TCHO, TG, LDL, AST, ALT and CK levels among the 4 groups. The expressions of CD34 and CD31 protein in glomerulus, and the expressions of eNOS and VEGF mRNA in renal tissue were higher in atorvastin treatment groups than those in CRF model group. The expression of ET-1 mRNA in renal tissue were lower in atorvastin treatment groups than those in CRF model group. CONCLUSION: It might be through promoting renovation of glomerulus capillary endothelium and improving function of glomerular endothelial cells that atorvastin ameliorates renal pathological injury and renal function in rats with chronic renal failure. Copyright 2012 by the Chinese Pharmaceutical Association.
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Objective: To explore the correlation between endothelia cells activation and cytokines (ET-1, NO) levels in patients with pulmonary hypertension (PH), and to discuss their roles in the development of PH. Methods: Twenty patients with simple ventricular septal defect (VSD) were chosen as controls, and 30 patients with PH were studied. Plasma levels of ET-1 and NO were measured by radioimmunoassay or colorimetric method. Before cardiopulmonary bypass was established, the specimens from right lung were fixed with formaldehyde solution, embedded with paraffin and stained by SP immunohistochemistry. Intercellular adhesion molecule-1 (ICAM-1) expression was measured through the determination of the light density with computer imaging technology. Results: Compared with that of the patients with simple VSD, the light density of ICAM-1 and plasma level of ET-1 increased in patients with PH; but plasma level of NO decreased (P<0.05). Positive correlation was observed between ICAM-1 and ET-1/NO (P<0.05). Conclusion: Endothelia cells activation and imbalance of ET-1/NO might play an important role in the development of PH.
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Objective To explore the correlation between endothelia cells activation and cytokines (ET-1, NO) levels in patients with pulmonary hypertension (PH), and to discuss their roles in the development of PH. Methods Twenty patients with simple ventricular septal defect (VSD) were chosen as controls, and 30 patients with PH were studied. Plasma levels of ET-1 and NO were measured by radioimmunoassay or colorimetric method. Before cardiopulmonary bypass was established, the specimens from right lung were fixed with formaldehyde solution, embedded with paraffin and stained by SP immunohistochemistry. Intercellular adhesion molecule-1 (ICAM-1) expression was measured through the determination of the light density with computer imaging technology. Results Compared with that of the patients with simple VSD, the light density of ICAM-1 and plasma level of ET-1 increased in patients with PH; but plasma level of NO decreased (P<0.05). Positive correlation was observed between ICAM-1 and ET-1/NO (P<0.05). Conclusion Endothelia cells activation and imbalance of ET-1/NO might play an important role in the development of PH.
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0.05) respectively. CONCLUSION: DMR can relax isolated rabbit pulmonary artery on the basis of endothelium-dependent and may involve in nitric oxide (NO), but is not related to blockage of receptor-operated and voltage-dependent calcium channels.