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Objective To verify enzyme activity inhibition of a novel histone deacetylase inhibitor ( HDACi ) JZ005 using an HDACi chemiluminescence detection kit and a cell-based screening model .Methods The plasmid with p21 gene promoter elements and luciferase reporter gene was transfected into human embryonic kidney cells 293 , and the stable transfectants were established by G418 screening.Enzyme activity inhibition of JZ005 on histone deacetylases (HDACs) was verified by the HDACi chemiluminescence detection kit and the cell-based screening model .A well-known HDACi , tri-chostatin A ( TSA) was used as the positive control .MTT assay was used to detect the protection of rat H 9c2 myocardial cells suffering from CoCl 2-induced hypoxia and treated with different concentrations of JZ 005 .The expression of acetylated histone H3 protein of normal and CoCl 2-induced hypoxia H9c2 cells before and after JZ005 treatment was assayed by West-ern blotting while the effect of drug administration on apoptosis was detected by flow cytometry ( FCM) .Results An HDA-Ci cell-based screening system targeting the p21 gene promoter was ranging established .The JZ005, a HDACi, markedly suppressed the activity of HDACs by more than 50%with the concentration ranging from 50 to 400 μmol/L.JZ005 signifi-cantly protected H9c2 cells from hypoxia injury .Cell viability was increased by 38.33%,56.00% and 35.20% compared with control,accompanied by an enhanced acetylation level of histone H 3.JZ005(25,50 and 100 μmol/L) treatment sig-nificantly decreased the number of apoptotic cells (6.63%,10.56% and 8.89%) compared to control group (12.89%). Conclusion An HDACi cell-based screening system is successfully established .JZ005 effectively protects myocardial cells against hypoxia injury while enhancing the acetylation level of histone H 3.Our results indicate that JZ005 might be developed as a potential drug for hypoxia treatment .
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Objective To discuss the effect of the combination detection of cardiac troponin I (cTnI) and homocysteine(Hcy) for increasing the diagnosis and treatment offects of non-ST elevation myocardial infarction (NSTEMI) .Methods The levels of cTnI and Hcy were detected in 47 patients with NSTEMI(NSTEMI group) before and after therapy and 63 healthy individuals(control group) .The detection results were performed the statistical analysis for verifying their value to judge the diagnostic and treatment effect of NSTEMI .Results The levels of cTnI and Hcy were (2 .37 ± 0 .65)ng/mL and(19 .23 ± 2 .94)μmol/L in the NSTEMI group ,which were significantly higher than(0 .33 ± 0 .14)ng/mL and(10 .62 ± 3 .27)μmol/L in the control group ,the differences showing statistical significance (P< 0 .05);the sensitivities of single cTnI and Hcy were 95 .74% and 85 .11% respectively ,and their specificities were 85 .71% and 90 .48% respectively ;the sensitivity and sepecificity of cTnI and Hcy combination detection were risen to 97 .87% and 98 .41% respectively ;after therapy ,the cTnI and Hcy levels in the NSTEMI group were significantly lowered and close to the normal levels .Conclusion The combination detection of cTnI and Hcy can not only be used for the diagno-sis of NSTEMI ,but also has the important significance to the judgment of the therapeutical effect of NSTEMI .
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Objective To examine the anticancer effect of a novel histone deacetylase inhibitor (HDACi), JZ004, on colon cancer cells HCT-8 and HT-29, and to investigate the molecular mechanisms of proliferation inhibition and apoptosis induction of cancer cells treated by JZ 004.Methods Colon cancer cells were treated with a series of concentrations of JZ004 .MTT assay was used to detect the proliferation of cancer cells .The cell cycle distribution and apoptosis were deter-mined by flow cytometry .Rhodamine 123 and DCFH-DA were applied to detect the mitochondrial membrane potential (ΔΨm) and reactive oxygen species ( ROS) production.The protein expressions of acetyl-histone H3, p21, cyclin-dependent kinase(CDK)4, Bcl-2, Mcl-1 and Bax were assayed by Western blotting .Results JZ004 was found to inhibit proliferation and induce apoptosis of colon cancer cells in a time-and dose-dependent manner , accompanied by a dose-dependent hyperacetylation of histone H3.JZ004 induced the cancer cell arrest in G 0/G1 phase by increasing the expres-sion level of p21 while CDK4 was downregulated .JZ004 also increased cellular ROS production and reduced ΔΨm by regu-lating the expressions of Bcl-2 family proteins .Conclusion As a novel HDACi , JZ004 effectively inhibits proliferation and increases ROS production to induce apoptosis of colon cancer cells .The results indicate that JZ004 is a potential compound to be developed as an anti-colon cancer agent for clinic application .
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Objective: Soy products may cause excessive intestinal gas because of soybean oligosaccharides . The effect of recombinant ?-galactosidase on eliminating mouse flatus was observed. Methods:The mouse model of flatulence was set up by ig raffinose and stachyose and the flatulence was investigated by measuring intestinal flatulent volume. Oligosaccharides were examined by TLC test and soybean protein was examined by SDS-PAGE. Results: Raffinose and stachyose can result in mouse flatus and recombinant ?-galactosidase can eliminate it without any effect on soybean protein. Conclusion: Recombinant ?-galactosidase can eliminate flatus, and be used as food additive.