OBJECTIVETo evaluate the effect of concentrated growth factor extract (CGFe) on the proliferation and differentiation of MC3T3-E1 osteoblasts attached to sandblasted and acid etched titanium surfaces.

METHODSTrials were divided into experimental and control groups. The experimental group used a-MEM that contained CGFe (10% FBS), whereas the control group only used a-MEM (10% FBS). MTT assay was employed to detect the number of osteoblasts on the first, third, and fifth days. Alkaline phosphatase (ALP) activity and scanning electron microscope (SEM) were used to detect the osteoblast differentiations on the third and fifth days and to observe the osteoblast extensions on titanium surfaces for 12 h, respectively. Meanwhile, the levels of the osteogenetic biomarkers Runt-related transcription factor-2 (Runx2) and Osterix (Osx) on the third and seventh days were quantified via real-time polymerase chain reaction (PCR).

RESULTSMTT assay indicated that on the first, third, and fifth days, the absorbance in the experimental group significantly increased than that in the control group (P < 0.05). ALP activity: on the third and fifth days, the absorbance of the experimental group was significantly higher than that of the control group (P < 0.05). SEM: at 12 h, the extension of the experimental group cells was larger than that of the control group. Real-time PCR: given the standardization in the group, the gene expression level of the control group on the third day was 1, and the Runx2 and Osx gene expressions in the experimental group were larger than those of the con- trol group.

CONCLUSIONCGFe can efficiently stimulate the proliferation, differentiation and extension of MC3T3-E1 cells.

Cell Differentiation ; Cell Line ; Cell Proliferation ; Core Binding Factor Alpha 1 Subunit ; Intercellular Signaling Peptides and Proteins ; Osteoblasts ; Osteogenesis ; Titanium

Fatigue not only affects people′s lives and work, but also causes diseases .Numerous studies have shown that some phytochemicals can promote body energy metabolism and regeneration , improve physical condition , resist fatigue growth and accelerate fatigue alleviation .This paper reviewed the recent research progress , domestic and overseas , in the anti-fatigue effect of phytochemicals .

Objective To investigate the effect of ski gene in migration process of astrocytes in rats. Methods Astrocytes were obtained from rats' cerebral cortex and cultured in vitro. siRNA targeting ski gene and negative control sequences were prepared. The ski-siRNA group, siRNA negative control group and untreated group were set in this experiment. The specific siRNA targeting ski gene was transfected into astrocytes with Lipofectamine?RNAiMAX Reagent. Then the ski protein levels were determined with Western blotting. After transfec-tion, the changes in migration of astrocytes were measured with wound scratch assay and Transwell migration assay. Results Western blot-ting showed that the expression of ski protein was significantly lower in the ski-siRNA group than in the siRNA negative control group and untreated group (F=132.957, P<0.001). Transwell migration assay showed that the number of astrocytes crossing through chambers was less in the ski-siRNA group than in the siRNA negative control group and untreated group (F>47.197, P<0.05). Wound scratch assay showed that the wound healing rate was lower in the ski-siRNA group than in the control group one, two, three, four and five days after transfection (F>69.187, P<0.001). Conclusion Ski knocked down by siRNA could inhibit the migration ability of astrocytes. It is a reminding that ski may take part in the migration process of astrocytes, and moreover, ski may play an important role in the formation of glial scar.

Objective To evaluate the effect of concentrated growth factor extract (CGFe) on the proliferation and differen-tiation of MC3T3-E1 osteoblasts attached to sandblasted and acid etched titanium surfaces. Methods Trials were divided into experimental and control groups. The experimental group used α-MEM that contained CGFe (10% FBS), whereas the control group only used α-MEM (10% FBS). MTT assay was employed to detect the number of osteoblasts on the first, third, and fifth days. Alkaline phosphatase (ALP) activity and scanning electron microscope (SEM) were used to detect the osteoblast differentiations on the third and fifth days and to observe the osteoblast extensions on titanium surfaces for 12 h, respectively. Meanwhile, the levels of the osteogenetic biomarkers Runt-related transcription factor-2 (Runx2) and Osterix (Osx) on the third and seventh days were quantified via real-time polymerase chain reaction (PCR). Results MTT assay indicated that on the first, third, and fifth days, the absorbance in the experimental group significantly increased than that in the control group (P<0.05). ALP activity: on the third and fifth days, the absorbance of the experimental group was significantly higher than that of the control group (P<0.05). SEM: at 12 h, the extension of the experimental group cells was larger than that of the control group. Real-time PCR: given the standardization in the group, the gene expression level of the control group on the third day was 1, and the Runx2 and Osx gene expressions in the experimental group were larger than those of the con-trol group. Conclusion CGFe can efficiently stimulate the proliferation, differentiation and extension of MC3T3-E1 cells.

Objective:To investigate the effect of IKKε on autophagy during abdominal aortic aneurysm formation in mice.Methods:We stimulated ApoE -/- mice and ApoE -/-IKKε -/- mice with AngⅡ and saline for 28 days. Metaboilic levels and aortic diameter of ApoE -/- mice and ApoE -/-IKKε -/- mice were measured. The arterial fibrosis of mice was detected by Masson staining and HE staining, mitochondrial reactive oxides were detected by fluorescence assay, the expression levels of autophagy factors LC3B and Beclin-1 were detected by immunohistochemistry, and the protein level of LC3B was detected by Western blot. Results:There was no significant difference in metaboilic levels between ApoE -/- mice and ApoE -/- IKKε -/- mice. However, the aortic diameterf ApoE -/-IKKε -/- mice was significantly less than that of ApoE -/- mice. The fibrosis level of ApoE -/-IKKε -/- mice was significantly lower than that of ApoE -/- mice. Furthermore, ROS in ApoE -/-IKKε -/- mice was lower than that in ApoE -/- mice. In addition, immunohistochemical and western blot showed that the expression levels of LC3B and Beclin-1 in ApoE -/-IKKε -/- mice were significantly lower than in ApoE -/- mice. Conclusion:IKKε -/- deficiency can significantly inhibit autophagy, thus reducing the development of abdominal aortic aneurysm in mice.