Calreticulin promotes migration and invasion of nasopharyngeal carcino-ma cells by inducing cell EMT / 中国病理生理杂志

AIM:To observe the expression of calreticulin(CRT)in nasopharyngeal carcinoma tissues,ana-lyze the significance of clinical pathology and the influence on epithelial -mesencymal transition(EMT)of CNE2 cells. METHODS:The expression of calreticulin was detected by immunohistochemistry in 52 nasopharyngeal carcinoma and 57 nasopharyngeal benign tissues,and the significance of clinical pathology was evaluated.The calreticulin gene-specific small interfering RNA was constructed,and then was transfected into the NPC cell line CNE 2 using the cationic liposome meth-od.The effect of CRT on the morphological changes of the CNE 2 cells was observed under light microscope.The effect of CRT on the cell migration and invasion abilities of the CNE 2 cells was detected by Transwell migration and invasion assays. The expression of EMT-related proteins E-cadherin,vimentin,transforming growth factor(TGF)-βand matrix metallopro-teinase(MMP)-9 in the CNE2 cells was determined by Western blot.RESULTS:The positive expression rate of CRT in the benign lesion tissues was 19.29%(11/57),which was significantly increased in the nasopharyngeal carcinoma tissues as 82.69%(43/52).The expression rate of CRT was positively correlated with the stage of nasopharyngeal carcinoma and lymph node metastasis(P<0.05).Knockdown of CRT expression made the CNE 2 cells showing a spindle shape to a flat, cobblestone-like epithelial state change,arranged more compact,and the migration and invasion abilities were significantly decreased(P<0.05).Knockdown of CRT expression resulted in significant increase in the protein expression of E -cadhe-rin,and the decreases in the protein expression of vimentin, TGF-βand MMP-9 in the CNE2 cells(P<0.05).CON-CLUSION:Calreticulin expression in nasopharyngeal carcinoma is significantly higher and positively correlated with naso -pharyngeal carcinoma stage and lymph node metastasis.Calreticulin promotes cell migration and invasion of nasopharyngeal carcinoma CNE2 cells by inducing EMT.

Effects of proton pump inhibitors on in-stent restenosis in patients receiving clopidogrel: a retrospective analysis / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To determine the effect of proton pump inhibitor (PPI) on in-stent restenosis (ISR) in patients receiving clopidogrel therapy.</p><p><b>METHODS</b>Total 439 patients underwent percutaneous coronary intervention (PCI) were enrolled in the study,including 250 post-PCI patients discharged on clopidogrel alone and 189 patients discharged on clopidogrel with PPI. The in-stent restenosis (ISR) ratio of the patients in these two groups were observed.</p><p><b>RESULTS</b>During a mean follow-up period of (13 ± 5.9) months, the post-PCI patients discharged on concomitant clopidogrel-PPI therapy had higher risk of ISR than those discharged on clopidogrel alone (19.6% Compared with 8%, P<0.001).</p><p><b>CONCLUSION</b>Concomitant use of clopidogrel and PPI after hospital discharge would increase the risk of ISR for post-PCI patients.</p>

The effect of chelerythrine on the hypertrophy of cardiac myocytes of neonatal rats induced by different glucose levels and its mechanism / 药学学报

The purpose of this study is to investigate the effect of chelerythrine on the hypertrophy of cardiomyocytes of neonatal rats induced by different glucose levels and its mechanism. Using cultured neonatal ventricular myocytes as a model, groups were divided as: control (5 mmol x L(-1)); high glucose level (10, 15, 20, and 25.5 mmol x L(-1)); high glucose level (25.5 mmol x L(-1)) add different concentrations of chelerythrine (1 and 8 micromol x L(-1)); and control glucose level (5 mmol x L(-1)) add different concentrations of chelerythrine (1 and 8 micromol x L(-1)). Different groups of cardiomyocytes after adding corresponding treat factors were cultured for 48 hours. Cardiomyocytes' diameters and protein level were measured and the expression of PKC-alpha, PKC-beta2, p-PKC-alpha, and p-PKC-beta2 were measured by Western blotting. Compared with control group, neonatal myocytes cultured in high glucose levels showed increased cellular volumes, protein level and expression of PKC-alpha, PKC-beta2, p-PKC-alpha, p-PKC-beta2. When chelerythrine was added, cellular volumes, protein level and expression of PKC-alpha, PKC-beta2, p-PKC-alpha, p-PKC-beta2 were significantly reduced. But in 1 micromol x L(-1) chelerythrine group, the expression of PKC-beta2 was not significantly reduced. The result suggested that chelerythrine can reverse the hypertrophy induced by different glucose levels on the cardiac myocytes, it may have protective effect against diabetic cardiomyopathy via PKC passageway.

Changes of endothelial progenitor cells from peripheral blood in patients with coronary heart diseases / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate alterations of endothelial progenitor cells (EPCs) from peripheral blood in patients with coronary heart diseases.</p><p><b>METHODS</b>Twenty patients with coronary heart diseases (CHD) and 20 matched control subjects were included in the study. Total mononuclear cells (MNCs) were isolated from peripheral blood by Ficoll density gradient centrifugation, and then the cells were plated on fibronectin-coated culture dishes. After cultured for 7 days, attached cells were cytochemically analyzed. EPCs were characterized as adherent cells double positive for DiLDL-uptake and lectin-binding by laser scanning confocal microscope with direct fluorescent staining. EPCs proliferation and migration were measured by MTT assay and modified Boyden chamber assay, respectively. EPCs adhesion assay was performed by replating on fibronectin-coated dishes, then adherent cells were counted.</p><p><b>RESULTS</b>The number of EPCs was significantly reduced in patients with CHD compared with that of age-matched control subjects (31.8+/-7.7 compared with 59.5 +/-10.6 EPCs/x 200 field; P<0.05). In addition, the functional activity of EPCs such as proliferation, migration and adhesive capacity was also impaired in patients with CHD.</p><p><b>CONCLUSION</b>EPCs number and functional activity are significantly decreased in patients with CHD.</p>

Effects of puerarin on number and activity of endothelial progenitor cells from peripheral blood / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate whether puerarin can augment endothelial progenitor cells (EPCs) numbers, promote EPC proliferation, migration and adhesion.</p><p><b>METHOD</b>Total mononuclear cells (MNCs) were isolated from peripheral blood by Ficoll density gradient centrifugation, and then the cells were plated on fibronectin-coated culture dishes. After 7 days culture, attached cells were stimulated with puerarin (to make a series of final concentrations: 0. 1, 0.5, 1, 3 mmol x L(-1)) or vehicle control for the respective time points (6, 12, 24, 48 h). EPCs were characterized as adherent cells double positive for DiLDL-uptake and lectin binding by direct fluorescent staining under a laser scanning confocal microscope. EPCs proliferation, migration and in vitro vasculogenesis activity were assayed with MT assay, modified Boyden chamber assay and in vitro vasculogenesis kit, respectively. EPCs adhesion assay was performed by replating those on fibronectin-coated dishes, then adherent cells were counted.</p><p><b>RESULT</b>Incubation of isolated human MNCs with puerarin dose increased the number of EPCs, maximum at 3 mmol x L(-1), 24 hours (approximately 1-fold increase, P < 0.01). In addition, puerarin also promoted EPC proliferative, migratory, adhesive and in vitro vasculogenesis capacity.</p><p><b>CONCLUSION</b>Puerarin can augment the number of EPCs with enhanced functional activity.</p>

Effects of Ginkgo biloba extract on number and activity of endothelial progenitor cells from peripheral blood / 药学学报

<p><b>AIM</b>To investigate whether Ginkgo biloba extract can augment endothelial progenitor cell (EPC) number, and promote EPC proliferation, migration and adhesion.</p><p><b>METHODS</b>Total mononuclear cells (MNCs) were isolated from peripheral blood by Ficoll density gradient centrifugation, and then the cells were plated on fibronectin-coated culture dishes. After 7 days of culture, attached cells were stimulated with Ginkgo biloba extract (10, 25 and 50 mg x L(-1)) or vehicle control for the respective time points (6, 12, 24 and 48 h). EPC were characterized as adherent cells double positive for DiLDL-uptake and lectin binding by direct fluorescent staining under a laser scanning confocal microscope. EPC were further documented by demonstrating the expression of CD34, VEGFR-2 and AC133 with flow cytometry. EPC proliferation, migration and in vitro vasculogenesis activity were assayed with MTT assay, modified Boyden chamber assay and in vitro vasculogenesis kit, respectively. EPCs adhesion assay was performed by replating MNCs on fibronectin-coated dishes, and then counting adherent cells.</p><p><b>RESULTS</b>Incubation of isolated human MNCs with Ginkgo biloba extract increased the number of EPC, maximum at 25 mg x L(-1), 24 hours (approximately 1-fold increase, P < 0.01). In addition, Ginkgo biloba extract promotes EPC proliferative, migratory, adhesive and in vitro vasculogenesis capacity.</p><p><b>CONCLUSION</b>Ginkgo biloba may promote EPC augmentation and enhance its functional activity.</p>

Statins contribute to enhancement of the number and the function of endothelial progenitor cells from peripheral blood / 生理学报

The aim of the present study was to investigate whether fluvastatin augments the number of endothelial progenitor cells (EPCs), and promotes EPCs proliferation, migration and adhesion. Total mononuclear cells (MNCs) were isolated from peripheral blood by Ficoll density gradient centrifugation. The cells were then plated on fibronectin-coated culture dishes. After being cultured for 7 d, the attached cells were stimulated with fluvastatin (final concentrations: 0.01, 0.1, 1, 10 micromol/L), simvastatin (1 micromol/L) or a vehicle for the respective time points (6, 12, 24 and 48 h). EPCs were characterized as adherent cells double positive for DiLDL-uptake and lectin binding by direct fluorescent staining under a laser scanning confocal microscope. EPCs were further documented by demonstrating the expression of KDR, VEGFR-2 and AC133 with flow cytometry. EPCs proliferation, migration and in vitro vasculogenesis activity were assayed by MTT assay, modified Boyden chamber assay and in vitro vasculogenesis kit, respectively. EPCs adhesion assay was performed by replating it on fibronectin-coated dishes, and the adherent cells were then counted. In addition, we also studied EPCs culture assay of peripheral blood from fluvastatin-treated animals in vivo. Incubation of isolated human MNCs with fluvastatin dose- and time-dependently increased the number of EPCs, while reached the maximum 24 h after the administration at 1 micromol/L, (2.5-fold increase, P<0.05). Moreover, treatment of rats with fluvastatins elevated the number of EPCs (3-fold increase, P<0.05), thus extending the in vitro data. In addition, fluvastatin also promoted EPC proliferation, migration, adhesion and in vitro vasculogenesis in a concentration-dependent manner. The effects of fluvastatin on EPCs were compared with those of simvastatin at the same concentration (1 micromol/L), with a result of no statistical difference. The results of the present study define a novel mechanism of the action of statins: the augmentation of EPCs with enhanced functional activity.