OBJECTIVETo investigate the inhibitory effects of a novel histone deacetylases inhibitor FK228 on human colon cancer HCT-116 cells in vitro and in vivo, and evaluate its toxicity and side effects.

METHODSThe in vitro growth inhibitions of HCT-116 cells by different concentrations of FK228 and 5-Fu for 24, 48 and 72 h were assessed by CCK-8 assay. BALB/c nude mouse models of tumor xenografts were prepared by subcutaneous implantation of tumor tissue, and 4 mg/kg FK228 and 50 mg/kg 5-Fu were i.p. injected, respectively. The inhibitory effects on tumor growth, hematology, and liver and kidney function were evaluated.

RESULTSCCK-8 assay indicated that FK228 had an obvious growth inhibitory effect on HCT-116 cells in a dose- and time-dependent manner. The IC50 of FK228 in HCT-116 cells was 12.05 ng/ml for 48 h, while the IC50 of 5-Fu was 18.92 µg/ml. At 20 days after FK228 and 5-Fu treatment, the tumor volume of the FK228 group was (139.71 ± 44.54)mm(3), significantly lower than that of the 5-Fu group [(282.28 ± 58.81)mm(3)] and that of the model group [(520.65 ± 39.73)mm(3), P < 0.01 for both]. The average tumor weight was (0.07 ± 0.02)g in the FK228 group, significantly lower than that of the 5-Fu group [(0.20 ± 0.08)g, P < 0.01]. The tumor growth inhibition rate of the FK228 group was 73.2%, significantly higher than that of the 5-Fu group (45.8%, P < 0.01). The ALT levels of the FK228 and 5-Fu groups were significantly higher than that of the model group (P < 0.01). The BUN of 5-Fu group was significantly higher than that of the model group (P < 0.01), but the BUN of FK228 group was not significantly different from that of the blank and control groups (P > 0.05 for both). Routine blood test showed that WBC, RBC, Hb and PLT of the 5-Fu group were significantly lower than those of the model group (P < 0.05 for all), but only WBC of the FK228 group was significantly lower than that of the model group (P < 0.05). The pathological examination using HE staining revealed that in the FK228 group, there were fibrosis and inflammatory cell infiltration in the liver tissue, and mild edema of the renal tubules in the kidney. However, in the 5-Fu group there were extensive hepatocyte edema and necrosis in the liver, and evident deformation and necrosis of glomeruli and tubules, and tubular wall thinning in the kidney.

CONCLUSIONSThe results of this study indicate that FK228 can more effectively than 5-Fu inhibit the growth of HCT-116 cells in vitro and vivo, and without obvious toxic effect on the kidney and hematology. Its clinical value in colon cancer treatment deserves further investigation.

Alanine Transaminase ; blood ; Animals ; Antibiotics, Antineoplastic ; administration & dosage ; adverse effects ; pharmacology ; Antimetabolites, Antineoplastic ; pharmacology ; Blood Urea Nitrogen ; Cell Proliferation ; drug effects ; Depsipeptides ; administration & dosage ; adverse effects ; pharmacology ; Dose-Response Relationship, Drug ; Fluorouracil ; pharmacology ; HCT116 Cells ; Hematologic Tests ; Histone Deacetylase Inhibitors ; administration & dosage ; adverse effects ; pharmacology ; Humans ; Inhibitory Concentration 50 ; Kidney ; pathology ; Liver ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Random Allocation ; Tumor Burden ; drug effects ; Xenograft Model Antitumor Assays

Objective To study the association between Essen stroke risk score (ESRS) and coronary heart disease (CHD).Methods One hundred and forty-six patients who underwent coronary angiography in our hospital from July 1,2015 to December 31,2016 were divided into CHD group (n=105) with their stenosis ≥50％ and non-CHD group (n=41) with their stenosis ＜50％.Their clinical,laboratory and coronary angiography data were recorded and analyzed by univariate logistic regression analysis and multivariate logistic regression analysis respectively.ROC curve of ESRS was plotted for diagnosing CHD.Results The age was significantly older,the incidence of hypertension,diabetes mellitus and myocardial infarction,the ESRS and fasting blood glucose level were significantly higher in CHD group than in non-CHD group (P＜0.05,P＜0.01).The age,history of smoking and alcohol drinking,and the incidence of abnormal blood lipid,AF,other heart diseases,peripheral artery disease,ischemic stroke or the serum levels of TIA,TC,TG,LDL-C,HDL-C and uric acid were significantly different between CHD group and non-CHD group (P＞0.05).Multivariate logistic regression analysis showed that ESRS was an independent risk factor for CHD (OR=2.070,95％CI:1.473-2.908,P＜0.01).The area under the ROC curve of ESRS for diagnosing CHD was 0.743 (95％CI:0.651-0.834,P＜0.01).The Youden's index was the highest when the ESRS was 2 with a sensitivity of 80.95％ and a specificity of 58.54％.Conclusion ESRS is related with CHD and can thus predict the occurrence of CHD.It is necessary to make an overall assessment of CHD when the ESRS is ≥2.

T-cell internal antigen-1 (TIA-1) has roles in regulating alternative pre-mRNA splicing, mRNA translation, and stress granule (SG) formation in human cells. As an evolutionarily conserved response to environmental stress, SGs have been reported in various species. However, SG formation in chicken cells and the role of chicken TIA-1 (cTIA-1) in SG assembly has not been elucidated. In the present study, we cloned cTIA-1 and showed that it facilitates the assembly of canonical SGs in both human and chicken cells. Overexpression of the chicken prion-related domain (cPRD) of cTIA-1 that bore an N-terminal green fluorescent protein (GFP) tag (pntGFP-cPRD) or Flag tag (pFlag-cPRD) induced the production of typical SGs. However, C-terminal GFP-tagged cPRD induced notably large cytoplasmic granules that were devoid of endogenous G3BP1 and remained stable when exposed to cycloheximide, indicating that these were not typical SGs, and that the pntGFP tag influences cPRD localization. Finally, endogenous cTIA-1 was recruited to SGs in chicken cells and tissues under environmental stress. Taken together, our study provide evidence that cTIA-1 has a role in canonical SG formation in chicken cells and tissues. Our results also indicate that cPRD is necessary for SG aggregation.
Chickens ; Clone Cells ; Cycloheximide ; Cytoplasmic Granules ; Humans ; Protein Biosynthesis ; RNA Precursors ; RNA-Binding Proteins ; T-Lymphocytes

Bacterial ghost is intact envelope of Gram-negative bacteria, which is produced by the function of the lysis gene E from bacteriophage PhiX174. The expression of the lysis gene E is usually controlled by the thermosensitive lambdapL/pR-cI857 promoter. In this study, we described a mutation (T --> C) at the ninth nucleotide of the OR2 in the lambdapR promoter of the lambdapL/pR-cI857 system by overlap PCR. The bacteriolytic assay showed that the mutation in the lambdapL/pR-cI857 system enhanced the temperature of repressing the expression of gene E up to 37 degrees C. The lysis efficiency of altered lambdapR promoter in Escherichia coli DH5a and avian pathogenic E. coli DE17 was up to 99.9%. The expanded range of temperature will benefit for the production of bacterial ghost.
Bacteriolysis ; physiology ; Bacteriophage lambda ; genetics ; Base Sequence ; Cell Membrane ; physiology ; DNA, Bacterial ; analysis ; Escherichia coli ; genetics ; growth & development ; physiology ; virology ; Gene Expression Regulation ; genetics ; Molecular Sequence Data ; Mutation ; Promoter Regions, Genetic ; genetics ; Viral Proteins ; genetics ; metabolism

Objective To evaluate if caspase pathway was involved in streptomycin-induced cell apoptosis in cochlear hair cells. Methods F344 rats at postnatal day 3 or 4 were used for the study in cochlear organotypic cultures. The cochlear basilar membrane was micro-dissected out and cultured overnight, and then treated with 1 mmol/L streptomycin for 24 hours. Before the termination, the activity of caspase-8, 9 or 6 were detected with FAM-peptide-FMK labeled caspase-8, 9 or 6, respectively. The stereocilia and cuticular plate of hair cells were stained with TRITC conjugated phalloidin, and the nuclei were stained with Topro-3 DNA probe. The specimens were observed and photographed under confocal fluorescent microscope. Results Streptomycin with 1 nunol/L causes about 80% cochlear hair cells missing in the basal turn and 10% hair cell loss in the apex. After streptomycin treatment, the nuclear shrinkage and fragmentation were found in most cochlear hair cells, and the caspase-8, caspase-9 and caspase-6 were greatly activated. Conclusions Apoptosis is involved in the cochlear hair cells death induced by Streptomycin in vitro. The caspase activities in upstream and downstream are maybe the major apoptotic pathway.