Effects of HDAC Inhibitor Scriptaid on IM9 Cell Line and Its Mechanism / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To study the effect of HDAC inhibitor Scriptaid on multiple myeloma IM9 cells and preliminarily clarify the mechanism of Scriptaid-induced cell apoptosis.</p><p><b>METHODS</b>The cell viability, cell cycle and cell apoptosis were measured by CCK8 assay and flow cytometry respectively, the relative target gene expression levels were detected by RT-PCR, the effect of Scriptaid on p21 promoter activity was detected by using luciferase reporter assay.</p><p><b>RESULTS</b>Scriptaid inhibited IM9 cell viability in a dose-dependent manner. Scriptaid induced IM9 cell cycle arrest at G/M phase in a dose-dependent manner. Scriptaid triggered IM9 cell apoptosis was obviously, the mRNA levels of apoptosis-related proteins Caspase 9, Caspase 3 and PARP1 were also activated. The apoptosis-associated factors BAD, PTEN and p21 increased following treatment with different dose of Scriptaid, meanwhile, p21 promoter activity was also activated significantly.</p><p><b>CONCLUSION</b>HDAC inhibitor Scriptaid can promote IM9 cell apoptosis by transcriptional activation of p21 promoter in concentration-dependent manner.</p>

Relaxant effects of Aike Mixture on isolated bladder and prostatic urethral smooth muscle of rabbits / 中国结合医学杂志

<p><b>OBJECTIVE</b>To observe the relaxant effect of Aike Mixture (AKM) on isolated bladder and prostatic urethral smooth muscle of rabbits.</p><p><b>METHODS</b>The isolated bladder and prostatic urethral smooth muscle from male rabbits were placed in a Magnus bath and smooth muscle contraction was measured using a biological signal acquisition and analysis system. The effects of AKM in combination with methoxyamine, carbachol and CaCl2 on the contractile tension of muscle strips were determined by cumulative dosing.</p><p><b>RESULTS</b>AKM dose-dependently reduced contractile tension of bladder trigone smooth muscle (r=0.831, P<0.05), reduced contractile wave amplitude (r=0.837, P<0.05) and decreased contractile frequency (r=-0.917, P<0.01). AKM significantly inhibited the increases in smooth muscle contraction induced by methoxyamine, carbachol and CaCl2.</p><p><b>CONCLUSION</b>AKM dose-dependently inhibited the contraction of rabbit isolated bladder and prostatic urethral smooth muscle by antagonizing α1-adrenergic receptors and M-cholinergic receptors.</p>

Comparison of Postoperative Bleeding and Complications between Cemented and Non-cemented Bipolar Hemiarthroplasty in Treatment of Unstable Pertrochanteric Fracture / 대한고관절학회지

PURPOSE: In cases of patients who underwent bipolar hemiarthroplasty (BPHA) for treatment of a pertrochanteric fracture, we compared and analyzed the amount of blood loss and complications between a group using the cemented stem and a group using the cementless stem. MATERIALS AND METHODS: A total of 104 patients who underwent BPHA for treatment of a pertrochanteric fracture in our hospital for three years and 10 months (From January 2008 to October 2011) were included in this study. Among the 104 patients, 64 patients with a cemented stem were categorized into group 1, and the other 40 patients with an uncemented stem were categorized into group 2. Before surgery, the type of stem was determined by the bone quality of the proximal femur, which had been evaluated with a simple X-ray. Then, after surgery, the amount of blood loss and complications were compared between the two groups. RESULTS: Expected blood loss during the operation was 389.8 cc in group 1, and 395.3 cc in group 2(P=0.88). Postoperatively, average drained blood loss was 219.6 cc in group 1, and 338.1 cc in group 2. Cemented stem was associated with significantly less blood loss (P=0.004). The average operation time in group 1 and in group 2 was 96 minutes and 72 minutes. There was no significant difference in operating time (P=0.85). In addition, there was no difference in INR (International Normalized Ratio) and BMI (Body Mass Index) (P=0.28 and 0.08) regarding total amount of postoperatively drained blood loss. There was no occurrence of hypotensive shock or fatal pulmonary embolism in either group. Three cases of periprosthetic fracture occurred in group 2. CONCLUSION: Fewer occurrences of postoperative blood loss and fewer complications were observed in the cemented stem group than in the cementless stem group. Preoperative evaluation of bone quality and use of the cement stem for patients with poor bone quality may be a good treatment method that can help to reduce complications.

Evaluation of combination effects of 2- methoxyestradiol and methoxyamine on IUdRinduced radiosensitization in glioma spheroids

Glioblastoma is the most common and most malignant cancer of central nervous system. Targeted radiotherapy is an effective method toward its treatment. Iododeoxyuridine [IUdR] is a halogenated thymidine analogue known to be effective as a radiosensitizer in human cancer therapy. In this study we have evaluated the combination effects of 2-Methoxyestradiol, an inhibitor of hypoxia inducible factor 1 alpha [HIF-1 alpha] and Methoxyaminem, an inhibitor of base excision repair [BER] pathway on radiosensitization of IUdR in glioblastoma spheroid culture. The cytotoxic damages of DNA in U87MG cell line were compared using colony formation assay. Experiments were performed in large spheroids with a diameter of approximately 350micro m. Evaluation of the effects of IUdR with 2ME2 and MX pretreatment on spheroid cultured cell followed by ionizing irradiation showed more enhancemented [p

Recovery of Genes Epigenetically Altered by the Histone Deacetylase Inhibitor Scriptaid and Demethylating Agent 5-Azacytidine in Human Leukemia Cells

Histone deacetylation and demethylation are epigenetic mechanisms implicated in cancer. Studies regarding the role of modulation of gene expression utilizing the histone deacetylase inhibitor scriptaid and the demethylating agent 5-azacytidine in HL-60 leukemia cells have been limited. We studied the possibility of recovering epigenetically silenced genes by scriptaid and 5-azacytidine in human leukemia cells by DNA microarray analysis. The first group was leukemia cells that were cultured with 5-azacytidine. The second group was cultured with scriptaid. The other group was cultured with both agents. Two hundred seventy newly developed genes were expressed after the combination of 5-azacytidine and scriptaid. Twenty-nine genes were unchanged after the combination treatment of 5-azacytidine and scriptaid. Among the 270 genes, 13 genes were differed significantly from the control. HPGD , CPA3, CEACAM6, LOC653907, ETS1, RAB37, PMP22, FST, FOXC1, and CCL2 were up-regulated, and IGLL3, IGLL1, and ASS1 were down-regulated. Eleven genes associated with oncogenesis were found among the differentially expressed genes: ETS1, ASCL2, BTG2, BTG1, SLAMF6, CDKN2D, RRAS, RET, GIPC1, MAGEB, and RGL4. We report the results of our leukemia cell microarray profiles after epigenetic combination therapy with the hope that they are the starting point of selectively targeted epigenetic therapy.

Cytogenetic damages from iododeoxyuridine-induced radiosensitivity with and without methoxyamine in human glioblastoma spheroids

lododeoxyuridine-induced Radiosensitivityi [IUdR] is a halogenated thymidine analogue recognized to be effective in vitro and in vivo radioserisitizer in human cancers. It is reported that Methoxyamine [MX] potentiates DNA damages in cancer cells with blocking the repair pathway of lUdR damages. But studies, entirely, are restricted on monolayer culture cells from human colon cancer cells. Spheroids are 3D form of cells that aggregate and grow together which resemble in vivo tumor models in several aspects and the results of such studies can be extended to tumor in vivo. The aim of the current study was to evaluate DNA damages from IUdR and gamma rays with and without Methoxyamine in human Glioblastoma spheroids. The DNA induced damages in U87MG cell line were compared using alkaline comet assay method. Experiments were performed with two different sizes of spheroids [100omicrom and 300microm]. Evaluation of the effects of IUdR with and without MX pretreatment on spheroids following ionizing radiation showed that MX increased the cell damages of lUdR with and without irradiation in both diameters spheroids. The damages were further increased in 100microm compared with 300microm diameter. Comparisons of tail moments in spheroids with 100 and 300microm diameter showed that cell damages in larger spheroids, 300microm, are lesser than smaller one, 100microm. This could be due to existence of G[0] cells and cells with longer cycle which lUdR was less incorporated into them. Thus, decrease in lUdR radiosensitization and base wxcision repair [BER], results in reduction of MX activities. Using agents for Inhibiting the activities of proteins which are responsible for carrying the cells to G[0] may be beneficial in solving such problems

Diethylhydroxylamine given in vivo inhibits lipid peroxidation and lipofuscin formation in the nervous tissues of rat.

In vivo effects of diethylhydroxylamine (DEHA) on lipid peroxidation and lipofuscin formation in the nervous tissues of rat have been investigated. Rats were fed DEHA for 30, 60 and 90 days and lipid peroxidation levels and lipofuscin concentration measured in cerebellum, brain stem and spinal cord. Lipofuscin contents were also assessed histochemically. The results showed that the drug caused a significant reduction in lipid peroxidation level and lipofuscin concentration related to ageing.

Use of hydroxylamine in improving radio protection of a combination of 5-hydroxy L-tryptophan and a thiol compound (AET) in small mammals.

Radioprotective effectiveness has been evaluated by 30 day survival studies and protection to bone-marrow cells in mice after radiation exposure and this has been further established by 24 hr deoxycytidine excretion in urine of rats following 5 Gy whole body gamma irradiation and protection to superoxide dismutase enzyme in marrow cells and red blood corpuscles. Radioprotective effectiveness as well as the duration of radioprotection have been improved by the administration (ip) of hydroxylamine (20 mg/kg), a decarboxylase inhibitor, prior to the use of a combination of 5-hydroxy L-tryptophan (5-HTP, 70 mg/kg) and 2-aminoethylisothiuronium bromide hydrobromide (AET, 20 mg/kg) ip in small mammals before whole body gamma irradiation.

Interactions of methoxyamine with pyridoxal-5'-phosphate-Schiff's base at the active site of sheep liver serine hydroxymethyltransferase.

The mechanism of interaction of methoxyamine with sheep liver serine hydroxymethyltransferase (EC 2.1.2.1) (SHMT) was established by measuring changes in enzyme activity, visible absorption spectra, circular dichroism and fluorescence, and by evaluating the rate constant by stopped-flow spectrophotometry. Methoxyamine can be considered as the smallest substituted aminooxy derivative of hydroxylamine. It was a reversible noncompetitive inhibitor (Ki = 25 microM) of SHMT similar to O-amino-D-serine. Like in the interaction of O-amino-D-serine and aminooxyacetic acid, the first step in the reaction was very fast. This was evident by the rapid disappearance of the enzyme-Schiff base absorbance at 425 nm with a rate constant of 1.3 x 10(3) M-1 sec-1 and CD intensity at 430 nm. Concomitantly, there was an increase in absorbance at 388 nm (intermediate I). The next step in the reaction was the unimolecular conversion (1.1 x 10(-3) sec-1) of this intermediate to the final oxime absorbing at 325 nm. The identity of the oxime was established by its characteristic fluorescence emission at 460 nm when excited at 360 nm and by high performance liquid chromatography. These results highlight the specificity in interactions of aminooxy compounds with sheep liver serine hydroxymethyltransferase and that the carboxyl group of the inhibitors enhances the rate of the initial interaction with the enzyme.

Comparison of the microbiological turbidimetric and hydroxylamine colorimetric methods in stability study of cefadroxil monohydrate powder

Comparison between the chemical and microbiological methods for the analysis of cefadroxil in powder for reconstitution stored at different temperature was carried out. Results indicated that the values of the antibiotic percent obtained by chemical method differ than those obtained microbiologically of the same samples. Microbiological assay of stored samples at 25 gave information that the product is valid up to 30 months while the chemical method indicated that the same sample valid for 12 months only. Sample stored at 37C seemed valid after storage for 12 months using microbiological method but the chemical assay indicated that it was not valid after 6 months. Chemical and microbiological assay indicated that stored samples at 56C were not valid but there was difference between values obtained by chemical and microbiological methods. Microbiological assay of stored sample valid for 2 months but chemical assay showed them not valid at all. This indicated that the chemical method considered invalid for stability study and/or assessment of validity of product containing cefadroxil