ABSTRACT
BACKGROUND:Previous studies have shown that puerarin can inhibit the differentiation of osteoclasts,and the expression of Notch signaling pathway-related proteins such as Notch1,HES1,and Jagged1 is decreased.However,the specific mechanism of the Notch1 signaling pathway for the inhibition of osteoclast differentiation by puerarin is not clear. OBJECTIVE:To explore the effect of Notch signaling pathway on puerarin inhibiting the differentiation of mouse macrophage Raw264.7 into osteoclasts. METHODS:Raw264.7 cells were divided into seven groups for intervention culture.Blank control group was cultured in high-sugar DMEM medium;the osteoclast induction group was cultured in osteoclast induction medium;the puerarin intervention group was cultured with 50 μmol/L puerarin at the same time of osteoclast induction;Notch1 siRNA control group,Notch1 siRNA group,Notch1 overexpression control group and Notch1 overexpression group were transfected with Notch1 siRNA control sequence,Notch1 siRNA,Notch1 overexpression control plasmid and Notch1 overexpression plasmid,respectively,and then cultured with osteoclast induction medium and puerarin.The number and size of osteoclasts were observed by tartrate-resistant acid phosphatase staining,the skeleton formation of osteoclasts was observed by F-actin staining,and the gene expression level of osteoclast formation markers was detected by RT-PCR. RESULTS AND CONCLUSION:Tartrate-resistant acid phosphatase staining results showed that puerarin intervention could inhibit the generation of osteoclasts,Notch1 silencing could further reduce the number of osteoclasts,while the number of osteoclasts in the osteoclast-induced group increased significantly after Notch1 overexpression.The results of F-actin showed that Raw264.7 cells could form a well-defined F-actin ring after osteoclast induction.Puerarin intervention would inhibit the formation of cytoskeleton,and Notch1 silencing could aggravate the inhibitory effect of cytoskeleton formation,while Notch1 overexpression could alleviate this inhibitory effect of puerarin.RT-PCR results showed that puerarin could inhibit the mRNA expression levels of tartrate-resistant acid phosphatase,Cathepsin K and c-Fos,the expression of the above-mentioned three factors decreased significantly after Notch1 gene silencing,and Notch1 overexpression could upregulate the expression of these factors.These finding indicate that puerarin inhibits the differentiation of Raw264.7 cells into osteoclasts through the Notch signaling pathway.
ABSTRACT
BACKGROUND:Currently, literatures about autologous vein transplantation are few, and the research on the effect of different parts of autologous vein transplantation are not found yet. OBJECTIVE:To summarize the experiences of establishing the fistula using autologous vein transplantation so as to investigate the method of improving the success rate of surgery. METHODS:We analyzed retrospectively the data of 40 cases of establishing the fistula using autologous vein transplantation, and then compared the successful rate of autologous vein transplantation fistula, blood flow and operating time, thereby analyzing the influence of diabetes melitus on the successful rate of autologous vein transplantation fistula. RESULTS AND CONCLUSION:The successful rates of autologous vein transplantation fistula at different parts ranging from high to low were as folows: the cephalic vein, great saphenous vein, basilic vein and smal saphenous vein. Blood flow of the upper limb for vein transplantation fistula was obviously higher than that of the lower limb (P < 0.05). The operating time of autologous vein transplantation fistula was longer in the upper limbs than in the lower limbs (P < 0.01). For patients with diabetes melitus, the successful rate of autologous vein transplantation was markedly lower than those with no diabetes melitus (P < 0.01). For the hemodialysis patients with poor upper limb superficial vein, autologous vein transplantation is a better way of establishing the vascular access. Vein transplantation of the upper limbs is better than that of the lower limbs in success rate and operating time. Autologous vein transplantation fistula is not suitable for the patients with diabetes melitus.
ABSTRACT
AIM: To study the effects of tumor necrosis factor-α (TNF-α) on the transcriptional activity of nuclear factor-erythroid 2-related factor 2 (Nrf2) in pulmonary microvascular endothelial cells. METHODS: Rat pulmonary micro-vascular endothelial cells (PMVECs) were cultured by lung tissue block pasted methods, and identified immunocytochemically using Ⅷ factor-related antigen. The cells were treated with different doses TNF-α (prepared in serum-free medium) for 4 h. Subcellular localization and levels of Nrf2 in PMVECs were observed with immunocytochemical methods. Nuclear extract were obtained to assayed transcriptional activity of Nrf2 with EMSA. Total RNA were isolated to assay the mRNA expression of Nrf2 by RT-PCR. RESULTS: The protein level of Nrf2 in the nuclei and transcriptional activity increased dose-dependently in PMVECs after treated with TNF-α at concentrations of 2.5, 5.0 or 10.0 μg/L. However, the protein level of Nrf2 in nuclei and transcriptional activity decreased dose-dependently in PMVECs after treated with TNF-α at concentrations of 20 or 40 μg/L. No different mRNA expression of Nrf2 in PMVECs treated with TNF-α at all concentration above was observed. CONCLUSION: Transcriptional activity of Nrf2 increases in PMVECs treated with low or moderate doses of TNF-α and decreases in PMVECs treated with high doses of TNF-α.