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Objective Drug resistance is the main cause of chemotherapy failure in hepatocellular carcinoma (HCC), and thioredoxin reductase 1 (TXNRD1), as a major influencing factor for reactive oxygen species (ROS) metabolism, has been proven to be associated with the poor prognosis of patients with HCC. This study aims to explore the role of TXNRD1 in the mechanism of multidrug resistance in HCC. Methods BEL/FU cells in BEL-7402 cell line were selected as the multidrug-resistant cell line. The siRNA was used for the intervention of TXNRD1 expression; quantitative real-time PCR and Western blotting were used to measure the expression of TXNRD1; CCK-8 assay and flow cytometry were used to evaluate the effect of TXNRD1 on hepatocyte ROS accumulation, resistance to 5-fluorouracil (5-Fu) and doxorubicin (DOX), and apoptosis in vitro; a xenograft tumor model was established to investigate the effect of auranofin (AUR) on drug resistance in vivo. The two-independent-samples t test was used for comparison of continuous data between two groups. Results As a multidrug-resistant HCC cell line, BEL/Fu showed high mRNA and protein expression levels of TXNRD1 (both P < 0.05). Compared with 5-Fu or DOX treatment alone, the TXNRD1 inhibitor AUR combined with 5-Fu or DOX had had a significant reduction in the number of colony formation ( P < 0.01) and a significant increase in apoptosis ratio ( P < 0.001). The ROS scavenger N-acetylcysteine (NAC) significantly weakened the effect of TXNRD1 knockdown by siRNA on the drug resistance of BEL/Fu cells, and the application of NAC effectively reduced the apoptosis ratio of cells after siRNA interference ( P < 0.001). Animal experiments also confirmed that compared with the nude mice treated with 5-Fu alone, the nude mice treated with 5-Fu and AUR had a significantly lower tumor mass ( P < 0.001) and a significantly smaller tumor volume ( P < 0.001). Conclusion TXNRD1 plays an important role in the drug resistance of HCC, and inhibition of its level in cells can effectively improve drug resistance. As a TXNRD1 inhibitor, AUR has great application prospects in the multimodality therapy for HCC.
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Hepatocellular carcinoma is one of the common causes of tumor-related death, and it has high morbidity and mortality rates in China. Recent studies have shown that platelets are closely associated with the development of hepatocellular carcinoma. Literature review shows that platelets not only participate in hemostasis, but also act on liver cells and tumor microenvironment, promote the formation of new blood vessels, and participate in the development and progression of hepatocellular carcinoma as a cell mediator through immune response and other pathways. In addition, platelets and their derivatives can be used as potential therapeutic targets for hepatocellular carcinoma. Therefore, antiplatelet therapy is expected to become a new adjuvant strategy for the treatment of hepatocellular carcinoma, which has important clinical significance.
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It is known that extracorporeal shock wave (SW) may promote healing of fracture. A previous study reported that SW promoted human bone marrow stromal cells (hMSCs) towards osteoblasts in vitro. To study the osteogenesis ability of hMSCs treated by shock wave in porous hydroxyapatite (HA) in vivo, primary hMSCs of SW group and control group were cultured in the porous HA for 2 weeks and then implanted into subcutaneous sites of nude mouse. These implants were harvested and prepared for the biochemical analysis of alkaline phosphatase activity by AKP kit, histological analysis of decalcified and undecalcified sections and morphology by scan electric microscope (SEM), as well as osteocalcin mRNA expression by RT- PCR 4 weeks and 8 weeks after implantation. It showed that cells of SW and control group almost covered the rough surface of HA before implantation and the extracelluar matrix of SW group was abundant by SEM photomicrograph . The histological analysis and SEM photomicrograph showed active bone formation 4 weeks and 8 weeks after implantation, as well as tetracycline labeling under fluoroscopy analysis in SW group. Alkaline phosphatase in supernatants of the implants detected 4 weeks and 8 weeks after implantation in SW group was higher than in control group (P
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Objective To study the clinical characteristics,pathological type,diagnosis,treatment and follow-up result of the primary tumors of the sacrum.Methods The cases of 18 patients with primary tumors of the sacrum were treated by operation(12 cases with chordoma,1 case with osteosarcoma,5 cases with giant cell tumor of bone),patients with chordoma performed by radiotherapy after operation,and all patients had been followed up for 4～8 years.Results 15 patients were cured and 3 patients were recurred after operation.Conclusions Chordoma is the most frequent tumor in primary sacrum tumor.This kind of tumor should be treated by surgical and other assist methods.
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Objective To investigate the role of B7/CD28 costimulation pathway blockade with adenovirus-mediated CTLA4-Ig gene in macrophage and CD8~+T cell infiltration and cell apoptosis in murine liver transplantation. Methods Rat pairs were divided into three groups: SD-to-Wistar transplantation control group, CsA-treated group and CTLA4-Ig-treated group. IHC and TUNEL were used to analyze the expression of CTLA4-Ig gene in liver and immune cells infiltrate and cell apoptosis in liver grafts. Pathology was done on all harvested grafts. ResultsCTLA4-Ig gene expression was positive in the donor liver on day 7 after administering adenovirus-mediated CTLA4-Ig gene via vein, and remained positive until day 60 after liver transplantation. Infiltration of immune cells in CTLA4-Ig-treated group was less than that in rejection control group. the apoptotic index of rejection group on day 3,5,7 was significantly higher than those of CTLA4-Ig-treated. Conclusions CTLA4-Ig gene was constantly expressed in the donor liver after single intravenousely injection into rats using adenovirus as vector. Adenovirus-mediated CTLA4-Ig gene therapy can inhibit infiltration of immune cells and apoptosis in grafts, thus prolonging the survival of recipients.
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Objective To study the role of 1,25-dihydroxyvitamin D3 in preventing allograft acute rejection in rat orthotopic liver transplantation. Method Rats receiving orthotopic liver transplantation were divided into groupⅠ: syngenic control (Wistar-to-Wistar); GroupⅡ:acute rejection (SD-to-Wistar). GroupⅢ: acute rejection treated with cyclosporine; GroupⅣ: acute rejection treated with 1,25-(OH)_ 2 D_ 3 . Liver function, rejection index and IFN-? mRNA, IL-10 mRNA expression level were monitored on d1,5,7,15,30 posttransplantation. Results Survival of recipients in group Ⅳ was significantly prolonged (vs group Ⅱ, P
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ObjectiveTo investigate the mechanism by which cyclosporine A downregulates transcription of interferon-? gene after murine orthotopic liver transplantation.Methods Orthotopic murine liver transplantation model was employed and rats were divided into 3 groups. GroupⅠ: syngeneic control (Wistar-to-Wistar); GroupⅡ: acute rejection (SD-to-Wistar). GroupⅢ: acute rejection treated with cyclosporine A (SD-to-Wistar+CsA). EMSA was employed to analyze NFAT and NF-?B DNA-binding activity of splenocytes, RT-PCR was employed to analyze IFN-? mRNA transcription intragraft on 1,3,5,7,12 day posttransplant in each group, respectively. Histopathological examination was also performed for reference.Results In comparison to groupⅠ, NFAT and NF-?B DNA-binding activity of splenocytes in groupⅡincreased significantly( P
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AIM: To investigate the role of NFAT in cyclosporine A downregulating IFN-? gene transcription after liver transplantation. METHODS: Rat orthotopic liver transplantation model was employed and 3 groups of experiments were performed in this study. GroupⅠ: syngeneic control (Wistar-to-Wistar); GroupⅡ: acute rejection (SD-to-Wistar). GroupⅢ: acute rejection treated with cyclosporine A intramuscularly (SD-to-Wistar+CsA). EMSA and RT-PCR were used to analyze NFAT activity of splenocytes and IFN-? gene transcription intragraft of recipients with or without CsA treatment after liver transplantation. Histopathological assessment was also performed for reference. RESULTS: No noticeable rejection was detected in GroupⅠ while only low level of IFN-? mRNA transcription and faint NFAT activity were measured. In contrast, a marked rejection reaction was demonstrated from day3 postoperation in GroupⅡ. IFN-? mRNA transcription was significant and NFAT activity was intensive. In comparison to GroupⅡ, rejection grade in Group Ⅲ significantly decreased (P