Platelet factor 4 (PF4) induces cluster of differentiation 40 (CD40) expression in human aortic endothelial cells (HAECs) through the SIRT1/NF-κB/p65 signaling pathway.

PF4 is a pro-atherosclerotic molecule. Endothelial CD40, upon binding to its ligand CD40L, induces endothelial cell (EC) activation, which is a vital pathophysiological process in the initiation and progression of atherosclerosis. However, the relationship between PF4 and endothelial CD40 remains elusive. This study aims to investigate whether and how PF4 affects endothelial CD40 expression using primary HAECs. PF4 treatment down-regulated sirtuin 1 (SIRT1) expression but upregulated the expression of acetylated NF-κB p65 (Ac-p65) and CD40 in HAECs in a concentration- and time-dependent manner. Pretreatment with SIRT1 agonist (SRT1720 or RSV) or SIRT1-overexpressing lentivirus attenuated PF4-induced Ac-p65 and CD40 expression in HAECs, whereas preincubation with SIRT1 antagonist (NAM or EX527) or SIRT1 shRNA had the opposite effect. To investigate whether NF-κB/p65 signaling pathway modulates CD40 expression in PF4-treated HAECs, PDTC, a NF-κB inhibitor, and p65-shRNA were introduced. PDTC or p65-shRNA treatment down-regulated Ac-p65 expression in HAECs. PDTC or p65-shRNA preincubation suppressed CD40 expression in HAECs after PF4 treatment. To better determine whether SIRT1 regulates CD40 expression in PF4-treated HAECs via the NF-κB/p65 signaling pathway, p65-knockdown HAECs were preincubated with SIRT1 agonists before PF4 treatment. SIRT1 agonist preincubation further decreased CD40 expression in p65-knockdown HAECs treated with PF4. Moreover, PF4 treatment promoted p65 nuclear translocation in HAECs. The results of dual luciferase assay demonstrated that four NF-κB binding sites in the promoter of human CD40 gene were activated in PF4-treated HAECs. In conclusion, our findings suggest that PF4 treatment facilitates CD40 expression in HAECs through the SIRT1/NF-κB/p65 pathway.

Alteration of functional connectivity in patients with Alzheimer's disease revealed by resting-state functional magnetic resonance imaging.

The main symptom of patients with Alzheimer's disease is cognitive dysfunction. Alzheimer's disease is mainly diagnosed based on changes in brain structure. Functional connectivity reflects the synchrony of functional activities between non-adjacent brain regions, and changes in functional connectivity appear earlier than those in brain structure. In this study, we detected resting-state functional connectivity changes in patients with Alzheimer's disease to provide reference evidence for disease prediction. Functional magnetic resonance imaging data from patients with Alzheimer's disease were used to show whether particular white and gray matter areas had certain functional connectivity patterns and if these patterns changed with disease severity. In nine white and corresponding gray matter regions, correlations of normal cognition, early mild cognitive impairment, and late mild cognitive impairment with blood oxygen level-dependent signal time series were detected. Average correlation coefficient analysis indicated functional connectivity patterns between white and gray matter in the resting state of patients with Alzheimer's disease. Functional connectivity pattern variation correlated with disease severity, with some regions having relatively strong or weak correlations. We found that the correlation coefficients of five regions were 0.3-0.5 in patients with normal cognition and 0-0.2 in those developing Alzheimer's disease. Moreover, in the other four regions, the range increased to 0.45-0.7 with increasing cognitive impairment. In some white and gray matter areas, there were specific connectivity patterns. Changes in regional white and gray matter connectivity patterns may be used to predict Alzheimer's disease; however, detailed information on specific connectivity patterns is needed. All study data were obtained from the Alzheimer's Disease Neuroimaging Initiative Library of the Image and Data Archive Database.

Early initiation of argatroban therapy in the management of acute superior mesenteric venous thrombosis.

Acute superior mesenteric venous thrombosis (ASMVT) is an intractable disease with poor prognosis. Argatroban, a direct thrombin inhibitor, may be a novel anticoagulant method in the therapy of ASMVT. The aim of the present study was to assess the efficacy and safety of early argatroban therapy in ASMVT patients. The current retrospective study reviewed a consecutive series of ASMVT patients receiving early argatroban therapy during hospitalization between March 2013 and April 2014, with 18 ASMVT patients included in the study. Of these, 16 patients without hepatic dysfunction underwent anticoagulant therapy with argatroban with a mean dose of 1.57±0.34 µg/kg/min and a mean duration of 12.2±3.7 days, while their activated partial thromboplastin time (aPTT) was elevated to 1.95±0.26 times the baseline value. In addition, 2 hepatic dysfunction patients received therapy with a dose of 0.41 µg/kg/min and 0.46 µg/kg/min, and with aPTT of 1.68 and 1.62 times the baseline value, respectively. Overall, 94% (n=17) of the patients presented clinical improvement, while 88% (n=16) of patients presented partially or completely dissolved thrombus in contrast-enhanced computed tomography images. The incidence of surgery and bowel resection was 6% (excluding 1 case with intestinal necrosis detected on admission). Furthermore, 11% (n=2) of patients experienced a bleeding episode, however no major bleeding or mortality occurred during hospitalization. During the follow-up, the mortality and the recurrence rate were 6% and 11%, respectively. In conclusion, early initiation of argatroban treatment may be an effective and safe therapy in ASMVT, manifesting efficient resolution of the thrombus, rapid improvement of symptoms, low incidence of bowel resection and bleeding complication, and low mortality rate.

[The role of SHH pathway in PDGF-induced VSMC proliferation].

OBJECTIVE: To discuss the role of sonic hedgehog (SHH) pathway in PDGF-induced vascular smooth muscle cell proliferation. METHOD: Human vein VSMC were cultured in vitro. Laser confocal microscopy and Western blot were used to detect the expression of SHH pathway related proteins. The cell proliferation was evaluated by Ki-67 staining and BrdU incorporation after treatment by siRNA of Gli2 or shh pathway inhibitor cyclopamine. RESULTS: The results of laser confocal microscopy and Western blot showed that SHH pathway protein which including shh, Patched1 and Gli2 were activated in the PDGF-induced VSMC proliferation.BrdU incorporation assay and Ki-67 staining showed that the cell proliferation which induced by PDGF was inhibited by Gli2-siRNA and cyclopamine which can both block SHH pathway. CONCLUSION: SHH pathway play an important role in PDGF-induced VSMC proliferation.

Oncolytic adenovirus mediated Survivin RNA interference and 5-fluorouracil synergistically suppress the lymphatic metastasis of colorectal cancer.

Colorectal cancer is one of the leading malignancies in the world. The mortality is mainly caused by tumor metastasis. It is urgent to find new therapeutics against metastatic colorectal cancer. We constructed an adenovirus carrying a Survivin targeted shRNA, tested its effects alone or with 5-fluorouracil (5-FU) both in vitro and in a nude mouse xenograft model. Results showed that the recombinant adenovirus reduced the expression of Survivin effectively, and administration of virus together with 5-FU inhibited cancer cell metastasis both in vitro and in vivo at concentrations at which each agent alone was ineffective. We conclude Survivin targeted virotherapy together with 5-FU chemotherapy may be a promising treatment for metastatic colorectal cancer.

Effect of hypoxia-inducible factor 1-α on Survivin in colorectal cancer.

Colorectal cancer is a one of the most common malignancies. Hypoxia-inducible factor 1-α (HIF1-α) and Survivin play important roles in tumor development; however, the literature currently contains few reports on the relationship between them in colorectal cancer. In this study, we investigated the effect of HIF1-α on Survivin in colorectal cancer. Immunohistochemical staining was used to detect the expression of HIF1-α and Survivin in colorectal cancer tissue from 32 patients. Colon adenocarcinoma SW480 cells were cultured under normoxia and hypoxic conditions, and the expression of HIF1-α and Survivin was detected by RT-PCR and Western blotting. We also silenced HIF1-α in order to detect the expression of Survivin and cell apoptosis. In an in vivo xenograft tumor model, the effect of HIF1-α on cancer development and Survivin was evaluated by the measurment of tumor volume and immunohistochemical analysis. Analysis revealed that HIF1-α (75%) and Survivin (68.75%) were both overexpressed in colorectal cancer, and that their expression was correlated. They were also expressed in SW480 cells under conditions of normoxia, and exhibited a significant increase in expression under hypoxic conditions. The inhibition of HIF1-α by RNA interference decreased the expression of Survivin and led to the apoptosis of the SW480 cell line. In the in vivo xenograft tumor model, the expression of HIF1-α and Survivin was decreased in the siHIF1-α group, and the tumor volume (586.67±41.63 mm3) was much smaller than that in the negative interference (1374.67±85.87 mm3) and saline-treated (1382.80±28.42 mm3) groups. Our results indicate that HIF1-α is an important regulator of Survivin expression and has great potential capacity for cancer therapeutics.

Oncolytic adenovirus mediated Survivin knockdown by RNA interference suppresses human colorectal carcinoma growth in vitro and in vivo.

BACKGROUND: Colorectal cancer is a one of the most common alimentary malignancies. Survivin has been proved by many studies to be an ideal target for cancer gene therapy because of its strong anti-apoptotic effect. The reduction of Survivin expression by means of chemically synthesized small interfering RNA or small hairpin RNA expressed from plasmid and resulted growth inhibition of cancer cells had been proved by many studies including ours, but the transfection efficiency was not encouraging. So for the first time we constructed the Survivin shRNA into an oncolytic adenovirus, tested its effects on colorectal cancer cell lines and nude mice xenograft model. METHODS: In this study, we constructed an oncolytic adenovirus with a Survivin targeted small hairpin RNA and a reporter gene (ZD55-Sur-EGFP). The expression of Survivin mRNA and protein were analyzed by RT-PCR and western blot. The cell growth and apoptosis were tested by in vitro cytopathic assay, MTT assay and flow cytometry respectively. The effect of the constructed virus on xenograft model was evaluated by tumor volume and western blot analysis. RESULTS: ZD55-Sur-EGFP replicated in cancer cells specifically, reduced the expression of Survivin mRNA and protein expression effectively (P < 0.0001), induced cancer cell apoptosis and inhibited SW480 cell growth both in vitro and in vivo significantly. CONCLUSION: We conclude Survivin RNA interference combining with oncolytic adenovirus virotherapy to be a promising treatment for colorectal cancer.