Kinesin superfamily protein 4 (Kif4), a microtubule-based motor protein, has been shown to participate in a number of critical cellular processes, such as cell division, the intracellular transport of membranous vesicles and signal transduction. However, whether KIF4 regulates vascular endothelial growth factor (VEGF) receptor 1 (VEGFR1) expression remains unknown. Thus, in this study, in order to examine the effects of Kif4 on the expression of VEGFR1 in RAW264.7 monocytes/macrophages, Kif4 was silenced using siRNA. RT-qPCR, western blot analysis and ELISA were used to assess the expression of Kif4 and VEGFR1 up- and downstream signaling molecules, including VEGF-A, VEGFR1, soluble form of VEGFR1 (sVEGFR1), phosphorylated (p-)Akt and Akt. The silencing Kif4 inhibited the mRNA expression of VEGF (P<0.01) and p-Akt (P<0.05); however, the level of VEGF-A was increased (P<0.05) compared with the negative control siRNA-transfected group. The silencing of Kif4 decreased the VEGFR1 mRNA (P<0.05), VEGFR1 protein and sVEGFR1 levels in the cell supernatant (P<0.01). Following the application of insulin-like growth factor-1 (100 ng/ml), the specific agonist of PI3K/Akt in the Kif4 siRNA-transfected group, the VEGFR1 mRNA levels (P<0.001), the VEGFR1 protein levels and the sVEGFR1 (P<0.01) levels significantly increased; however, the levels of VEGF in the cell supernatant were decreased (P<0.05). Taken together, these findings suggest that Kif4 regulates the expression of VEGFR1 in RAW264.7 cells and that the PI3K/Akt pathway is involved in this process.
PURPOSE: To compare two different structural nanophase hydroxyapatite(HA) particles on the proliferation activity and functions of osteoblasts. METHODS: Primary culture of osteoblasts from rat calvaria was established and then cultured on the coatings of different size of nano particles (group I : 20-40nm,group II :70-100nm), the HA coatings without nano-particles was used as control group. Proliferation activity, protein content and synthesis of alkaline phosphatase (ALP) by osteoblasts were examined by MTT assay, coomassic brilliant blue method and PNPP test,and statistical significance was assessed by multiple comparison (q test, SNK) in one-way analysis of variance (ANOVA) with SPSS10.0 software package. RESULTS: Osteoblasts grew well on HA coatings. MTT assay demonstrated that there was significant difference between group I and group II at 6th day and 8th day (P0.05).At first half stage(5th day and 10th day) ALP activity test showed no significant difference between group I and group II (P0.05) and as the culture going on (15th day and 20th day), there was significant difference between the two groups (P0.05). Coomassic brilliant blue method showed that there was significant difference between group I and group II from 5th day to 20th day (P0.05). CONCLUSION: The diameter of nano-particles on the HA coatings could influence the proliferation activity and functions of the osteoblasts. The nano-particles of similar size with HA crystal in vivo showed better cytocompatibility. Supported by Research Fund of Science and Technology Bureau of Shandong Province(Grant No.032040105).
Objective:To study the effect of bone morphogenetic proteins-2 (ad-hBMP-2) gene therapy on mandibular fracture union.Methods: An animal model of mandibular fracture was established in 16 rabbits which were divided randomly into experimental and control group.100μl ad-hBMP-2 and normal saline were injected respectively into the breaking end of fracture 3 days after operation.All animals were killed 4 weeks after operation and the samples were examined by X-ray and histology.Results: The rate of the new bone formation and the density of the rebirth bone trabecula of the experimental group were superior to the control.Conclusion:ad-hBMP-2 gene therapy can accelerate bone formation during mandibular fracture in rabbits.
To investigate the effects of ischemia/reperfusion on rat submandibular glands without denervation and the possible protective effects of ischemia preconditioning on the glands that experienced ischemia/reperfusion, in‐situ ischemia/reperfusion and ischemia preconditioning experimental models of submandibular glands of healthy male Wistar rats were conducted. For ischemia/reperfusion groups, the glands were subjected to 90 min of ischemia without denervation, followed by 1, 12, 24, or 72 h of reperfusion. Ischemia preconditioning was achieved by 3 min of ischemia following 3 min of reperfusion, performed three times before ischemia/reperfusion. Salivary secretion, histological changes, alterations of tight junctions, myeloperoxidase activity, cellular apoptosis, and reactive oxygen species levels were detected. In ischemia/reperfusion glands, rising acute‐inflammation responses, reduced tight‐junction width, and increased myeloperoxidase activity, reactive oxygen species levels, and apoptotic cell numbers were observed, along with secretory dysfunction, especially at 1 and 12 h post‐reperfusion, which seemed to gradually return to normal by 72 h post‐reperfusion. In contrast, ischemia preconditioning showed the potential to ameliorate the injury‐stress responses caused by ischemia/reperfusion. Our study revealed that ischemia/reperfusion could cause a series of injury‐stress responses and ultimately lead to hyposecretion, independently of the parasympathetic nerve supply, which might play an important role in the early‐phase dysfunction of the transplanted glands. Ischemia preconditioning could protect the involved glands and improve ischemia/reperfusion‐induced hyposecretion.
Objective The present study aimed to explore the clinical significance of neutrophils infiltration and carcinoembryonic antigen related cell adhesion molecule 1 (CEACAM1) expression in the tongue squamous cell carcinoma (TSCC), and to probe the possible relationship between them. Materials and Methods Tissue microarray and immunohistochemistry were used to detect neutrophils density and CEACAM1 expression in 74 cases of primary TSCC specimens and 17 cases of corresponding peritumoral tissues. The relationship of CEACAM1 expression and neutrophils density with clinicopathologic parameters and cancer-related survival of TSCC patients were evaluated. The correlation between CEACAM1 expression and neutrophils density was also evaluated. Real-time quantitative transcription polymerase chain reaction (qRT-PCR) was used to explore the possible molecular mechanisms between CEACAM1 expression and neutrophils infiltration. Results Immunohistochemistry evaluation revealed that there was more neutrophils infiltration in TSCC tissues than in peritumoral tissues. High neutrophil density was associated with LN metastasis (P = 0.01), higher clinical stage (P = 0.037) and tumor recurrence (P = 0.024). CEACAM1 overexpression was also associated with lymph node metastasis (P = 0.000) and higher clinical stage (P = 0.001). Survival analysis revealed that both neutrophils infiltration and CEACAM1 overexpression were associated with poorer cancer-related survival of TSCC patients (P<0.05), and neutrophils infiltration was an independent prognostic factor for TSCC (P<0.05). Furthermore, overexpression of CEACAM1 was correlated with more neutrophils infiltration in TSCC tissues (P<0.01). qRT-PCR results showed that CEACAM1-4L can upregulate the mRNA expression of IL-8 and CXCL-6, which were strong chemotactic factors of neutrophils. Conclusion Our results demonstrated that more neutrophils infiltration and overexpression of CEACAM1 were associated with poor clinical outcomes in TSCC tissues. Overexpression of CEACAM1 on tumor cells correlated with more neutrophils infiltration to some extent through upregulating mRNA expression of IL-8 and CXCL-6.
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