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Objective:To investigate the effect and mechanism of Toll-like receptor 2 (TLR2) on cognitive function in obese mice.Methods:Male C57BL/6J and TLR2 knockout mice were divided into control group, obesity group, TLR2 knockout group, and TLR2 knockout obesity group according to standard diet or high-fat diet. After 16 weeks, water maze experiments were performed to test the learning and memory ability of mice in each group. The body weight and blood lipid biochemical indexes of the mice in each group were measured. Immunohistochemical analys was used to detect amyloid-β (Aβ) protein expression in the hippocampus. Western blotting was used to detect the expression of nuclear factor-κB (NF-κB) p65, phosphorylated (p-) NF-κB p65, low density lipoprotein receptor-related protein 1 (LRP1), and Aβ protein.Results:Compared with the standard control group, the expressions of TLR2, NF-κB p65, p-NF-κB p65, and Aβ protein in obesity group were significantly increased, LRP1 protein expression level was reduced, and the learning and memory ability of mice was significantly reduced. Compared with the obese group, the expression levels of NF-κB p65, p-NF-κB p65, and Aβ protein in the TLR2 gene knockout obesity group decreased, while the expression level of LRP1 protein increased. The memory and learning ability of these mice was significantly improved.Conclusion:TLR2 deficiency may improve the cognitive function of obese mice by inhibiting the TLR2/NF-κB pathway. The mechanism may be related to the up-regulation of LRP1 protein expression.
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Objective To investigate the antitumor activity of the procaspase-3 activator SM-1 in BGC-823 cells in vivo and in vitro and the mechanisms.Methods The inhibitory effects of SM-1 on proliferation of BGC-823 cells were evaluated using MTT method, the cell apoptosis rate was detected by flow cytometry, and the expression of caspase-3 protein and procaspase-3 mRNA was detected by Western blotting and RT-PCR, respectively.SM-1 Antitumor activity was evaluated using the xenograft of BGC-823 cells in nude mice.Results SM-1 effectively inhibited the proliferation in vitro and in-duced apoptosis of BGC-823 cells in a dose-dependent manner.After treatment with SM-1 for 48 h, the protein expression levels of caspase-3 and mRNA expression levels of procaspase-3 were increased.SM-1 significantly inhibited growth of BGC-823 xenograft tumor at the 300 mg/kg dose and the inhibition rate was 56.3%(P<0.05).Conclusion SM-1 can significantly inhibit the tumor growth of BGC-823 cells in vivo and in vitro.The mechanism is possibly related to the activation of procaspase-3 and induced apoptosis of tumor cells.
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Objective To establish a simple and useful kidney or bladder orthotopic tumor model used in preclinical pharmacodynamic evaluation.Methods Mouse model of orthotopic renal cancer were established by subrenal capsule implantation.After aspirating urine and irrigating bladder with PBS,the bladder urothelium was slightly impaired to establish the orthotopic bladder tumor model.Then, B-Ultrasound and H&E staining were used to confirm the availability.Results Tumors could be seen 2 weeks after surgery, accompanied by body mass loss of the mice.H&E staining showed that the tumor cells acted as infiltrative growth.The growth of tumor was inhibited by NTX in vivo, the tumor mass inhibitory rate of the KCC-853 orthotopic tumor model was 57.5% of 60 mg/kg NTX treatment and 48.8% in the T24 orthotopic tumor model of 30 mg/kg NTX treatment.Conclusion Our methods for establishing the orthotopic kidney or bladder tumor model are simple and practical.The results indicate that nitroxoline has potential antitumor activity.
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Aim To investigate the effect of TP on the expression of macrophages inflammatory protein ( MIP-1α) . Methods Total RNA of mouse Ana-1 cells and tumor associated macrophages were extracted, and MIP-1α mRNA was detected by RT-PCR. Mouse S180-xenografts were established by injecting S180 cells subcutaneously into the double abdominal flanks of the mice. The postoperative residual tumor models were generated in the right abdominal tumors when tumors grew into 250 mm3 . Animals were treated with TP or CTX, and tumor tissues were separated and MIP-1α was detected by immunohistochemistry. Results There was no significant difference of the expression of MIP-1α between Ana-1 cells and TAMs. TP couldn’ t affect MIP-1αexpression in Ana-1 cells while it signifi-cantly decrease MIP-1α expression in TAMs in a dose-dependent manner. TP significantly decreased MIP-1αexpression of tumor tissue compared with control group. Conclusions MIP-1α will be a new target of TP anti-cancer. Simple cell line tests in vitro couldn’ t reveal the real state in vivo.
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Objective To investigate the combination effect of cetuximab and irradiation on colorectal carcinoma CL187 cell line and underlying molecular mechanism.Methods CL187 cells with or without cetuximab treatment were irradiated by 0,4 and 8 Gy X-rays,then cell death percentage was determined by MTT 24 and 48 h post-irradiation.Clone forming assay was used to evaluate the cell reproliferation ability.Cell cycle distribution,apoptosis,and necrosis were analyzed by flow cytometry.Western blot was used to detect the protein expressions of DNA-PKcs,Ku70 and Ku80.Results The cetuximab enhanced the percentage of radiation-induced cell death,while descreased the cloning formation capacity and increased radiosenvtivity (t =-6.14、-6.53,P ＜0.05).The SER of cetuximab on CL187 cell line approached to 1.38.In addition,cetuximab also increased radiation-induced G0/G1 phase arrest (t=-4.64,P＜0.05) and the percentage of apoptosis and necrosis (t=-9.16,P ＜0.05),but it descreased the expression levels of DNA-PKcs,Ku70 and Ku80 proteins.Conclusions The cetuximab treatment might enhance the inhibitory effect of irradiation on colorectal carcinoma CL187 cell line by influencing cell cycle distribution,cell apoptosis,and the expression of DNA repair proteins.