ABSTRACT
Objective:To evaluate the effect and mechanism of davallia mariesil flavones (DMF) improving osteoporosis via Notch signaling pathway.Methods:(1) 120 cases patients with osteoporosis in our hospital from January 2016 to January 2017 were analyzed,and divided into experiment group (60 cases) and control group (60 cases) using digital random grouping method.Experiment group was treated with DMF combined with D-calfor.Control group was treated with D-calfor only.After treatment, the levels of serum calcium,serum phosphor,TNF-α,MCP-1 and IL-6 in serum were detected to evaluate the clinical efficacy.(2) Bone marrow stromal cells were separated and cultivate.NC group:DMEM(contain 10% FBS).RA group:RA(0.4 mmol/L).DMF+RA group:DMEM(contain DMF)+RA(0.4 mmol/L).Jaggedl+RA group:Jaggedl(1 000 μg/L)+RA(0.4 mmol/L).Jaggedl+DMF+RA group:Jaggedl(1 000 μg/L)+DMEM(contain DMF)+RA(0.4 mmol/L).DAPT+RA group:DAPT(16 μmol/L)+RA(0.4 mmol/L). DAPT+DMF+RA group:DAPT(16 μmol/L)+DMEM(contain DMF)+RA(0.4 mmol/L).Western blotting assays and PCR were performed to assess mRNA and protein levels of Notch-1,Hes-1.Results: (1) In clinical study,the effective rate in treatment group was obviously higher than control group (91.67%>76.67%,P<0.05).The levels of serum calcium and serum phosphor in the experiment group was higher than in the control group (P<0.05).The levels of TNF-α,MCP-1 and IL-6 in the experiment group was lower than in the control group (P<0.05).(2) In experimental study,compared with the RA group,the expressions of Notch-1,Hes-1 mRNA and protein were upregulated in Jaggedl+RA group,but were downregulated in DAPT+RA group,DMF+RA group (P<0.05). Compared with the Jaggedl+RA group,the expressions of Notch-1,Hes-1 mRNA and protein were downregulated in Jaggedl+DMF+RA group (P<0.05).Compared with the DAPT+RA group,the expressions of Notch-1,Hes-1 mRNA and protein were downregulated in DAPT+DMF+RA group (P<0.05).Conclusion:DMF could improve the condition of osteoporosis.The mechanism may be associated with inhibiting the Notch signaling pathway by DMF.
ABSTRACT
Brown algae are well known as a source of biologically active compounds, especially those having antioxidant activities, such as phlorotannins. In this study we examined the antioxidant activities of crude phlorotannins extracts (CPEs) obtained from Sargassum hemiphyllum (SH) and fractionated according to the molecular weights. When CPEs were administrated at a dose of 30 mg/kg to Kunming mice pre-treated with carbon tetrachloride (CCl4), the levels of oxidative stress indicators in the liver, kidney and brain were significantly reduced in vivo. All the components of various molecular weight fractions of CPEs exhibited greater scavenging capacities in clearing hydroxyl free radical and superoxide anion than the positive controls gallic acid, vitamin C and vitamin E. Particularly, the components greater than 30 kD obtained from ethyl acetate phase showed the highest antioxidant capacities. These results indicated that SH is a potential source for extracting phlorotannins, the algal antioxidant compounds.
Subject(s)
Animals , Male , Mice , Antioxidants , Pharmacology , Ascorbic Acid , Pharmacology , Brain , Metabolism , Pathology , Carbon Tetrachloride , Toxicity , Carbon Tetrachloride Poisoning , Drug Therapy , Metabolism , Pathology , Chemical Fractionation , Methods , Gallic Acid , Pharmacology , Hydroxyl Radical , Metabolism , Kidney , Metabolism , Pathology , Liquid-Liquid Extraction , Methods , Liver , Metabolism , Pathology , Mice, Inbred Strains , Oxidation-Reduction , Oxidative Stress , Phaeophyceae , Chemistry , Sargassum , Chemistry , Superoxides , Metabolism , Tannins , Pharmacology , Vitamin E , PharmacologyABSTRACT
<p><b>OBJECTIVE</b>To establish a method for diagnosis of freshwater drowning by amplifying gyrB and 16S rRNA genes of Aeromonas hydrophila using PCR coupled with capillary electrophoresis (CE).</p><p><b>METHODS</b>DNA samples were extracted from human, 18 planktons (including Candida albicans, Aeromonas hydrophila, and 16 species of algae), and 30 cases of tissue samples (including the lung, liver, and kidney, all examined with microwave digestion-vacuum filtration-automated scanning electron microscopy) from human cadavers, including 28 freshwater drowning victims and 2 with natural death. The DNA samples were amplified with the primer AH (for gyrB gene) and primer Ah (for 16S rRNA gene), and the products were analyzed with CE.</p><p><b>RESULTS</b>PCR amplification followed by CE yielded negative results for DNA of human, Candida albicans and 16 species of algae, whereas a positive result was found for Aeromonas hydrophila DNA with PCR products of 195 bp (with primer AH) and 350 bp (with primer Ah). In the 28 drowning cases, the detection rates of Aeromonas hydrophila using primer AH were 96.4% in the lung tissue, 71.4% in the liver tissue, and 60.7% in the kidney, as compared with the rates of 75.0%, 42.9%, and 32.1% using primer Ah, respectively. The positive rates for Aeromonas hydrophila in the organs of the drowning victims were 82.1% and 53.6% with primer AH and primer Ah, respectively. The detection showed negative results in the 2 cases of natural deaths. The two primers produced significantly different detection rates of Aeromonas hydrophila (P<0.05).</p><p><b>CONCLUSION</b>PCR coupled with CE for detecting gyrB gene of Aeromonas hydrophila has a high sensitivity in assisting a diagnosis of freshwater drowning. Detection of both the gyrB gene and 16S rRNA gene of Aeromonas hydrophila can yield more convincing evidence of the diagnosis of freshwater drowning.</p>
ABSTRACT
A novel of fibrinolytic protein has been separated and purified by ammonium sulfate fractionation, DEAE-cellulose and SephadexG-75 Column chromatography from the tissue of the female Eupolyphaga sinensis in the paper. The protein showed an apparent molecular weight of 41.3 kD on sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. In addition, it includes 10.5% sugar. Its specific activity to hydrolyze fibrin was 547.86 u/mg. The enzyme activity was inhibited by Mg2+, Ca2+, protein inhibitors, such as 8mol/L urea and 1% beta-mercaptoethanol, and serine protease inhibitor such as phenylmethanesulfonyl fluoride (PMSF), but wasn't inhibited by Na+, K+ and ethylenediaminotetraacetic acid (EDTA). The protein was stable under 40 degrees C and it's optimal temperature was also 40 degrees C. It's optimal pH was 8.0. It showed a different way between the activity and UK when they degrade the plasminogen. Based on all the messages the protein can be suggested to be a novel fibrinolytic protein. There have been no such component of fiberinolytic enzyme from Eupolyphaga sinensis walker reported yet.