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Objective@#To investigate the effects of miRNA on the expression of paraoxonase 1 (PON1) and its clinical application in the patients with nonalcoholic steatohepatitis (NASH). @*Methods@#Bioinformatics methods were used to analyze and predict PON1 related regulation on miRNA. PON1 luciferase reporter gene vectors were constructed and the activity of dual luciferase was analyzed. The up/down-regulated levels of miRNA in HepG2 cells of different groups were detected by real-time fluorescence quantitative PCR (qRT-PCR), and the levels of PON1 protein in HepG2 cells were detected by western blot. The levels of miR140-5p in the serum of healthy people and NASH patients were also analyzed by qRT-PCR. @*Results@#According to the prediction of TargetScan database, miR140-5p may bind complementarily to the end of PON13′-UTR. The analysis for the activity of dual luciferase reporter gene showed that miR-140-5p mimic significantly downregulated the fluorescence of wild type PON1 vector (P＜0.01). The results of qRT-PCR demonstrated that miR-140-5p mimic group showed high overexpression (P＜0.01) compared with the normal cell control group and the negative mimic control group, while miR-140-5p inhibitor group appeared corresponding low expression (P＜0.05). western blot results suggested that the transfection of miR140-5p mimic significantly down-regulated the expression of PON1 (P＜0.01) while miR140-5p inhibitor up-regulated this expression (P＜0.01). Compared with the healthy control group, the level of miR140-5p was decreased in the serum of NASH patients, and the difference was statistically significant (P＜0.01). @*Conclusion@#miR140-5p may be involved in the progression of nonalcoholic steatohepatitis through regulation for the posttranscriptional gene expression of PON1.
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BACKGROUND: In recent years, the immunoregulatory effects of mesenchymal stem cells (MSCs) can be used to induce immune tolerance. OBJECTIVE: To evaluate the safety and feasibility of human umbilical cord-derived MSCs (hUC-MSCs) in liver transplantation patients. METHODS: hUC-MSCs were cultured and identified. After approved by the Medical Ethics Committee, a total of 50 patients were randomly divided into experimental group and control group according to the proportion of 1:1. In the experimental group, hUC-MSCs were perfused by the portal vein during the operation and infused into the jugular vein on the 3rd day after the operation. The injection dose was 1×106/kg (prepared as 50 mL of cell suspension). Both groups received standard immunosuppressive regimens. Blood biochemistry and immune function indicators were detected preoperatively and at postoperative days 3, 7, months 1, 2, 3, 6, 12. Acute and chronic rejection rates, incidence of infection, and incidence of transplantation-related complications were recorded. RESULTS AND CONCLUSION: (1) At 3 and 7 days after the operation, the percentage of peripheral blood CD4+CD25+ cells (regulatory T cells) in the experimental group was significantly higher than that in the control group (P 0.05). (3) The incidence of abnormal liver function in the experimental group was significantly lower than that in the control group (P 0.05). Overall, the intravenous infusion of hUC-MSCs is safe and feasible in liver transplantation patients, which in early stage can promote the the proliferation and activation of CD4+CD25+ cells (regulatory T cells), reduce the percentage of CD4+ cells (helper/inducer T cells) and lower the ratio of CD4+/CD8+ T cells, thereby improving the immune status in liver transplantation patients.
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Objective Bevacizumab ( BM ) is an angiogenesis inhibitor widely used in cancer therapy, but its off-target effect of proteinuria may lead to discontinuation of treatment.This study was to explore the mechanisms of BM inducing proteinuria in mice. Methods Twenty-four healthy mice were randomly divided into four groups, saline control, low-dose BM, medium-dose BM, and high-dose BM, treated by injection of normal saline and BM at 10, 35, and 60 mg per kg of the body weight, respectively, though the tail vein.At 4 weeks after injection, 24-hour urine was collected to determine the total urine protein and blood obtained from the eyeballs for biochemical analysis.Then all the mice were sacrificed and the kidneys harvested for observation of pathologic changes in the glomeruli as well as for immunohistochemistry, Western blotting, and real-time PCR analysis. Results Compared with normal saline,BM obviously elevated the level of 24-hour urine protein, with statistically significant differences between the control and the medium-and high-dose BM groups (0.23 ±0.02 vs 1.14 ±0.13 and 1.43 ±0.10, P0.05).No significant differences were observed among the four groups in the levels of Cr, BUN, AST and ALT (P>0.05).Under the optical microscope, the kidneys showed normal structures in the control group, no signifi-cant pathologic changes in the low-dose BM, and vacuolus-like alteration with atrophic glomerular endothelial cells in the medium-and high-dose BM groups.Immunohistochemical analysis demonstrated the expressions of VEGF and podocin were moderately or strongly positive in the control and low-dose BM groups, by weakly positive or negative in the medium-and high-dose BM groups.Compared with the control group, the expression of the VEGF protein in the renal tissue was significantly decreased in the high-dose BM group (0.76 ±0.09 vs 0.39 ±0.05, P0.05), and the expression of the podocin protein was significantly reduced in the medium-dose BM (0.67 ±0.07 vs 0.43 ±0.10, P0.05).The mRNA expressions of VEGF and podocin were not significantly changed in the low-dose BM group as compared with the control (1.07 ±0.61 and 1.12 ±0.09 vs 1.23 ±0.25 and 1.17 ±0.19, P>0.05) but remarkably de-creased in the medium-dose (0.82 ±0.38 and 0.71 ±0.18) and high-dose BM groups and (0.47 ±0.64 and 0.42 ±0.09) groups (P<0.01). Conclusion Bevacizumab damages glomerular filtration membrane and induce proteinuria partially by down-regulating the protein and mRNA expressions of VEGF and podocin.
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Objective Antiangiogenesis therapy has been shown to prolong survival for patients with malignant tumor .However the present study has not been observed the clinical benefit of antiangiogenesis therapy combination with chemotherapy treated with gastric canc-er.Human recombinant vascular endothelial inhibition (endostar) as a multi-targeted anti-angiogenesis drug, the mechanism is different from other Antiangiogenesis drugs.It can block different pathways of signal transduction to inhibit angiogenesis .This study aimed to observe the effect of combined application of endostar and paclitaxel on biological behavior of gastric cancer cell lines . Methods MMT assay and Tr-answell invasion assay were respectively used to examine the inhibition rate of cell growth and invasion ability when cells were treated with va-rious concentrations of endostar and paclitaxel alone or in combination.The protein expressions of VEGF,MMP-2 and MMP-9 were examined by Western blot. Results Endostar or paclitaxel effectively inhibited the growth of MGC803 cells and the in vitro invasion of MGC803 cells in a concentration-dependent manner.The proliferation and invasion ability of combined treatment with endostar and paclitaxel was significantly lower than that of endostar or paclitaxel alone (P<0.05).Compared with con-trol group, the VEGF,MMP-2 and MMP-9 protein expressions were de-creased in experimental groups ( P <0.05).Compared with paclitaxel group, the VEGF, MMP-2 and MMP-9 protein expressions were relatively reduced in combination groups (P<0.05). Conclusion Endostar combined with paclitaxel can suppress the growth and invasion of MGC803 cells, and the decreasing VEGF , MMP-2 and MMP-9 expressions may be involved in the mechanism .
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Nowadays Chinese serious environment problems and the intensifying conflict between man and nature will affect not only Chinese economic and social development but also person's all-round development.Therefore,it is necessary to reveal the conflict between man and nature and the influence on person's all-round development as well as establish an effective mechanism for sustainable development.
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Objective To investigate paclitaxel cross resistance and provide information for clinical therapy by establishing a paclitaxel resistant human lung adenocarcinoma cell line.Method A paclitaxel resistant human lung adenocarcinoma cell line named with SPC-A1/Taxol was developed by intermittent exposure to gradually increasing concentration of taxol.Resistant index was calculated from median inhibitory concentration(IC50)which was evaluated by MTT assay.Results SPC-A1/Taxol cell line displayed high resistance to taxotere,vinorelbine,vincristine and doxorubicin.It also displayed median or low resistance to camptothecins and podophyllotoxins.No cross resistance was observed to antitumor platinum and antimetabolite drugs and elemene emusion.Conclusion SPC-A1/Taxol cell line is a typical multidrug-resistant cell line in vitro.It could be useful for studing the cross resistance and direct clinical medication.