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Objective To explore the effects and mechanism of high expression of ARNO (ARF nucleotide-binding-site opener) on the permeability of human renal glomerular endothelial cells (HRGECs) under high glucose conditions. Methods HRGECs were cultured in vitro, and ARNO expression and endothelial permeability were detected in high glucose time gradient (15, 30, 45 and 60 min) and concentration gradient (10, 20, 30 and 40 mmol/L) experiments, respectively. Cell lines of HRGECs with ARNO and Arf6 gene silencing were obtained by infecting HRGECs with ARNO siRNA and Arf6 siRNA recombinant lentivirus vectors, respectively. The effects of ARNO and Arf6 gene silencing on endothelial permeability were observed. Results In the time gradient experiments, compared with control group, ARNO expressions increased significantly in the groups with high glucose for 15, 30, 45 and 60 min (0.670±0.051, 0.960±0.106, 0.716±0.026, 0.531±0.030 vs. 0.242±0.029. P

Objective To validate the glomerular vascular lesions percentage score (GVLPS) system in renal vascular lesions of Henoch-Schönlein purpura nephritis (HSPN) in a single-center cohort. Methods Clinical and renal histopathological data of 140 patients with renal biopsy-proven HSPN, admitted in Xinqiao Hospital of Army Medical University from Jan. 2012 to Aug. 2015, were retrospectively analyzed and reclassified according to the GVLPS system. The GVLPS [number of glomeruli with endothelial cell proliferation, hyalinosis, cellulose necrosis, subendothelial hyaline deposits, cellular crescents, fibrous crescents, segmental sclerosis or global sclerosis/number of total obtained glomeruli×100%] was evaluated in each patient and classified into histopathological grades (HG): HG1 (

Objective To investigate the effect of over-expression of ROBO4 on permeability of human renal glomerular endothelial cells (HRGECs) in high glucose medium. Methods HRGECs infected with recombinant lentiviral vector ROBO4 were cultured in high glucose or low glucose medium in vitro. The protein levels of ROBO4 and ARF6 in each group were detected by Western blotting. The endothelial permeability was measured by the efflux of fluorescein isothiocyanate-dextran (FITC-Dextran) permeated through the monolayer endothelial cells using Transwell cell model system. The cell viability after lentivirus transfection was measured by CCK8 assay. Results The transfection rate of lentiviruses in HRGECs reached 80% 72h after, and obvious over-expression of ROBO4 protein was in transformed cells compared with the empty vector group (P

In this work, a 1600 tons forging machine was employed to conduct forging on the Ti-6Al-4V alloy. The microstructural and textural evolution of forged alloy were investigated by optical microscopy (OM), scanning electron microscopy (SEM), electron backscattered diffraction technique (EBSD) and X-ray diffraction (XRD). The results showed that the lamellar α phase in the Ti-6Al-4V alloy changed into a spherical morphology, and the spheroidization rate and the structure uniformity increased with the increasing forging times. The largest pole density in each incomplete pole figure of the α phase decreased gradually and the high-angle grain boundaries (HAGBs) gradually turned to the low-angle grain boundaries (LAGBs), which indicated that the texture was reduced during the forging process.

Abnormal angiogenesis plays a pathological role in diabetic nephropathy (DN), contributing to glomerular hypertrophy and microalbuminuria. Slit2/Robo1 signaling participates in angiogenesis in some pathological contexts, but whether it is involved in glomerular abnormal angiogenesis of early DN is unclear. The present study evaluated the effects of Slit2/Robo1 signaling pathway on angiogenesis of human renal glomerular endothelial cells (HRGECs) exposed to a diabetic-like environment or recombinant Slit2-N. To remove the effect of Slit2 derived from mesangial cells, human renal mesangial cells (HRMCs) grown in high glucose (HG) medium (33 mM) were transfected with Slit2 siRNA and then the HG-HRMCs-CM with Slit2 depletion was collected after 48 h. HRGECs were cultured in the HG-HRMCs-CM or recombinant Slit2-N for 0, 6, 12, 24, or 48 h. The mRNA and protein expressions of Slit2/Robo1, PI3K/Akt and HIF-1α/VEGF signaling pathways were detected by quantitative real-time PCR, western blotting, and ELISA, respectively. The CCK-8 cell proliferation assay, flow cytometry and the scratch wound-healing assay were used to assess cell proliferation, cycles, and migration, respectively. Matrigel was used to perform a tubule formation assay. Our results showed that the HG-HRMCs-CM with Slit2 depletion enhanced the activation of Slit2/Robo1, PI3K/Akt, and HIF-1α/VEGF signaling in HRGECs in time-dependent manner (0–24 h post-treatment). In addition, the HG-HRMCs-CM with Slit2 depletion significantly promoted HRGECs proliferation, migration, and tube formation. Pretreatment of HRGECs with Robo1 siRNA suppressed the activation of PI3K/Akt and HIF-1α/VEGF signaling and inhibited angiogenesis, whereas PI3K inhibitor suppressed HIF-1α/VEGF signaling, without influencing Robo1 expression. In the HRGECs treated with Slit2-N, Slit2-N time-dependently enhanced the activation of Robo1/PI3K/Akt/VEGF pathway but not HIF-1α activity, and promoted HRGECs proliferation, migration, and tube formation. The effects induced by Slit2 were also abolished by Robo1 siRNA and PI3K inhibitor. Taken together, our findings indicate that in a diabetic-like environment, in addition to mesangial cells, autocrine activation of Slit2/Robo1 signaling of HRGECs may contribute to angiogenesis of HRGECs through PI3K/Akt/VEGF pathway; therefore, Slit2/Robo1 signaling may be a potent therapeutic target for the treatment of abnormal angiogenesis in early DN and may have broad implications for the treatment of other diseases dependent on pathologic angiogenesis.