ABSTRACT
Objective To investigate the intervention effect of Xuanfuhua decoction on mice with nonalcoholic steatohepatitis (NASH) induced by high-fat, high-fructose, and high-cholesterol diet. Methods A total of 32 male C57/BL6J mice were randomly divided into normal group, model group, Xuanfuhua decoction group, and obeticholic acid group, with 8 mice in each group. Since week 24 of modeling using high-fat, high-fructose, and high-cholesterol diet, each group was given the corresponding drug for intervention at a dose of 14.19 g/kg by gavage for the Xuanfuhua decoction group and 10 mg/kg by gavage for the obeticholic acid group and a volume of 20 mL/kg for gavage, once a day for 6 consecutive weeks. HE staining, oil red O staining, Sirius Red staining, and Masson staining were used to observe the pathological changes, lipid deposition, and collagen deposition of liver tissue; related kits were used to measure the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), and glucose, as well as the content of TG and hydroxyproline (Hyp) in liver tissue; quantitative real-time PCR was used to measure the expression of genes associated with lipid metabolism, inflammation, and fibrosis in liver tissue; immunohistochemical staining was used to observe the positive expression of F4/80 and α-SMA in liver tissue. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t -test was used for further comparison between two groups. Results Compared with the normal group, the model group had significant increases in body weight, liver wet weight, and serum levels of AST, ALT, TC, TG, LDL-C and glucose (all P < 0.01). HE staining showed hepatocyte steatosis, a large number of fat vacuoles, hepatocyte ballooning degeneration, and inflammatory cell infiltration in liver tissue of the mice in the model group, and the model group had a significant increase in NAFLD activity score (NAS) compared with the normal group ( P < 0.01). Oil red O staining showed the deposition of a large number of red lipid droplets with different sizes in hepatocytes of the mice in the model group, and compared with the normal group, the model group had significant increases in the area percentage of oil red O staining and the content of TG in the liver ( P < 0.01). Sirius Red staining and Masson staining showed significant collagen fiber hyperplasia in the perisinusoidal area, the central vein, and the portal area in the model group, and the model group had a significant increase in the content of Hyp in liver tissue compared with the normal group ( P < 0.05). Compared with the model group, the Xuanfuhua decoction group had significant reductions in the serum levels of AST, ALT, TC, TG, LDL-C, and glucose (all P < 0.05), significant improvements in hepatic steatosis, inflammatory infiltration, lipid droplet deposition, and collagen fiber hyperplasia, and significant reductions in NAS score, area percentage of oil red O staining, and content of TG and Hyp in the liver (all P < 0.05). Compared with the normal group, the model group had significant increases in the mRNA expression levels of lipid metabolism-related genes (SREBP-1c, FASN, SCD-1, PPAR-γ, and CD36), inflammation-related genes (F4/80, TNF-α, CCL2, and CD11b), and the fibrosis-related gene α-SMA (all P < 0.05), and immunohistochemical staining showed significant increases in the positive expression of F4/80 and α-SMA ( P < 0.01). Compared with the model group, the Xuanfuhua decoction group had significant reductions in the mRNA expression levels of SREBP-1c, FASN, SCD-1, PPAR-γ, CD36, F4/80, TNF-α, CCL2, CD11b, and α-SMA in liver tissue (all P < 0.05), and immunohistochemical staining showed significant reductions in the positive expression of F4/80 and α-SMA ( P < 0.01). Conclusion Xuanfuhua decoction has a good intervention effect on mice with NASH induced by high fat, high fructose, and high-cholesterol diet and can significantly inhibit hepatic lipid deposition, inflammatory response, and liver fibrosis.
ABSTRACT
Objective:To investigate the expression change of blood plasma miRNA-210 (miR-210) in blood plasma for patients with esophageal squamous cell carcinoma (ESCC) before and after radiotherapy and its effect on cell proliferation, cycle, apoptosis of ESCC cells in vitro.Methods:The blood specimens from 22 patients with newly diagnosed ESCC (ESCC group) before and after radical radiotherapy between December 2013 and March 2015 in Linyi People's Hospital of Shandong Province as well as 15 healthy controls (the healthy control group) were collected. miRNA-21 (miR-21) was treated as the positive controls. The real-time polymerase chain reaction (RT-PCR) was used to detect the expression level of miR-210 and miR-21 in blood plasma before and after radiotherapy for patients, and the expression of miR-21 in ESCC Eca109 cells at normoxia and hypoxia time. The cell proliferation, cycle and apoptosis of Eca109 cells in untransfected group (the blank control group), miR-21 transfected mimics negative control RNA group (negative control group) and miR-210 transfected mimics RNA group (miR-210 group) were detected by using EdU cell proliferation assay and flow cytometry.Results:A total of 59 plasma samples from ESCC group and the healthy control group were collected. The relative expression level [median ( P25, P75)] of blood plasma miR-210 and miR-21 in ESCC group before radiotherapy was higher than that in the healthy control group, and the difference was statistically significant [4.04×10 -4 (2.06×10 -4, 6.68×10 -4) vs. 0.54×10 -4 (0.28×10 -4, 0.77×10 -4), 397.07×10 -4 (181.77×10 -4, 742.93×10 -4) vs. 127.43×10 -4 (21.97×10 -4, 184.65×10 -4); U value was 37.0, 49.0, respectively, all P < 0.01]. The expression level of miR-210 and miR-21 before radiotherapy in ESCC group was not related with tumor location and differentiation degree (all P > 0.05). After the radical radiotherapy one week later, the relative expression level of miR-210 and miR-21 in blood plasma of ESCC group was 65.33×10 -4 (22.15×10 -4, 160.87×10 -4), 437.23×10 -4 (327.18×10 -4, 749.09×10 -4), respectively, which was higher compared with that before radiotherapy ( U value was 32.0, 154.0, respectively, both P < 0.05), and the increase of miR-210 was more significant. The expression level of miR-210 after hypoxic cultured Eca109 cells for 12 h was increased compared with the normal oxygen with the peak value 12 h later, and the difference was statistically significant ( P < 0.01). EdU cell proliferation assay showed that the Eca109 cell proliferative activity after transfection of 24 h and 48 h in miR-210 group was decreased compared with the negative control group and the blank control group, and the difference was statistically significant (all P < 0.01). Flow cytometry analysis showed that the proportion of G 2/M phase of Eca109 cells in miR-210 group after transfection of 24 h was increased compared with the negative control group and the blank control group, and the difference was statistically significant ( P < 0.05). After transfection of 48 h, the increased proportion in G 2/M phase was more obvious ( P <0.01); there was no statistically difference in the apoptotic cell proportion among three groups after transfection of 24 h and 48 h (all P > 0.05). Conclusions:miR-210 is highly expressed in the blood plasma of ESCC patients, especially the significant increase in the expression level of miR-210 in blood plasma after radical radiotherapy. miR-210 is highly expressed in hypoxic ESCC Eca109 cells. The overexpression of miR-210 can inhibit cell proliferation and its mechanism may be related with cell arrest in G 2/M phase.
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The research and development and competition situation of global wearable blood pressure measuring device technologies covered in Thomson Innovation Patents Database and Patents Retrieval System of State Intellec-tual Property Office were analyzed by statistical analysis, clustering analysis, citation analysis, similarity matrix a-nalysis and literature analysis respectively with Thomson data analyzer, which showed that cuffless blood pressure measuring device technology is the hotspot.