To provide the profiles of metabolism of mitomycin C (MMC) by human liver microsomes in vitro, MMC was incubated with human liver microsomes, then the supernatant component was isolated and detected by HPLC. Types of metabolic enzymes were estimated by the effect of NADPH or dicumarol (DIC) on metabolism of MMC. Standard, reaction, background control (microsomes was inactivated), negative control (no NADPH), and inhibitor group (adding DIC) were assigned, the results were analyzed by Graphpad Prism 4. 0 software. Reaction group compared with background control and negative control groups, 3 NADPH-dependent absorption peaks were additionally isolated by HPLC after MMC were incubated with human liver microsomes. Their retention times were 10. 0, 14. 0, 14. 8 min ( named as Ml, M2, M3) , respectively. Their formation was kept as Sigmoidal dose-response and their Km were 0. 52 (95% CI, 0. 40 - 0.67) mmol x L(-1), 0. 81 (95% CI, 0. 59 - 1. 10) mmol x L(-1), 0. 54 (95% CI, 0. 41 -0. 71) mmol x L(-1) , respectively. The data indicated that the three absorption peaks isolated by HPLC were metabolites of MMC. DIC can inhibit formation of M2, it' s dose-effect fitted to Sigmoidal curve and it' s IC50 was 59. 68 (95% CI, 40. 66 - 87. 61) micromol x L(-1) , which indicated DT-diaphorase could take part in the formation of M2. MMC can be metabolized by human liver microsomes in vitro, and at least three metabolites of MMC could be isolated by HPLC in the experiment, further study showed DT-diaphorase participated in the formation of M2.
Antibiotics, Antineoplastic ; metabolism ; Chromatography, High Pressure Liquid ; methods ; Dicumarol ; pharmacology ; Dose-Response Relationship, Drug ; Enzyme Inhibitors ; pharmacology ; Humans ; Microsomes, Liver ; drug effects ; enzymology ; metabolism ; Mitomycin ; metabolism

OBJECTIVETo investigate the effect of early intervention by pravastatin with two different dosage on inflammatory factors and endothelial vasodilator function in patients with unstable angina (UA).

METHODS108 patients with UA were investigated consecutively and divided randomly into three groups (group 20 mg, n = 37; group 10 mg, n = 37; group control, n = 34). Blood samples were examined at admission and 4, 8 weeks after the therapy of pravastatin. Fourty patients of UA were chosen from those three groups (15, 15 and 10 cases respectively). The endothelium-dependent vasodilation and the function of vascular endothelium of them were measured. In the dosage of 20 mg pravastatin group non-endothelium-dependent vasodilation in brachial artery was also tested by ultrasound before and 8 weeks after the therapy. Cardiac events were followed up for 2 months.

RESULTS(1) The use of pravastatin in early admission period of UA could significantly reduce inflammatory factors and improve vascular endothelium function, which was more obviously in the group of 20 mg/d than in group of 10 mg/d. These benefits occurred in 4th week and more obviously in 8th week after the therapy. (2) The lipid lowering therapy in the early stage of admission (24 - 48 h) resulted in the reduction of cardiac events in the hospital.

CONCLUSIONThe use of pravastatin 20 mg/d seems better than that of 10 mg/d in all the fields as above in early admission period of UA patients.

Adult ; Aged ; Angina, Unstable ; drug therapy ; Anticholesteremic Agents ; administration & dosage ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Inpatients ; Male ; Middle Aged ; Pravastatin ; administration & dosage ; therapeutic use ; Prospective Studies

OBJECTIVETo investigate the effects of FTY720, a new immunosuppressive agent, on apoptosis and autophagy in multiple myeloma(MM) cell line U266 and to clarify its molecular mechanism.

METHODSU266 cells were treated with 0, 2.5, 5.0, 10.0 and 20.0 µmol/L FTY720 for 24 hours, and the cell viability was assayed by CCK-8 method. Then U266 cells were treated with 20.0 µmol/L FTY720 for 0, 2, 6 and 24 hours, the cell viability was tested. The apoptotic rates induced by different doses and time points of FTY720 were tested by flow cytometry separately. The expression of LC3B was detected by Western blot after U266 cells treated with different doses of FTY720 to see autophagy. U266 cells were treated with FTY720 ± Bafilomycin A1, an inhibitor of autophagy, for 24 hours, then the cell viability and apoptotic rates were tested. Meanwhile the expression of survivin, anti-apoptotic factors, were tested by Western blot.

RESULTSThe cell viability and the apoptotic rates were inhibited significantly by FTY720 (P < 0.05) in time-dependent and dose-dependent manner. The expression of LC3B-II increased significantly in a dose-dependent manner, it indicated that the autophagy was induced by FTY720. Bafilomycin A1 could rescue the cell viability and apoptotic rates in U266 cells treated with FTY720, and it could also rescue the expression of survivin decreased by FTY720.

CONCLUSIONSFTY720 can cause apoptosis and autophagy of U266 cells. The autophagy promote the apoptosis, which maybe due to the degradation of anti-apoptotic factors such as survivin or their upstream factors in lysosomes through autophagy.

Apoptosis ; drug effects ; Autophagy ; drug effects ; Cell Line, Tumor ; Fingolimod Hydrochloride ; Humans ; Multiple Myeloma ; pathology ; Propylene Glycols ; pharmacology ; Sphingosine ; analogs & derivatives ; pharmacology

This study was purpose to investigate the role of reactive oxygen species (ROS) in apoptosis and autophagy induced by FTY720 in multiple myeloma cell line U266. U266 cells were treated by different concentrations of FTY720 for 24 h, the apoptotic rates were detected by flow cytometry, and the expression of LC3B was detected by Western blot. The results indicated that apoptosis and autophagy were induced by FTY720 in U266 cells. Autophagy induced by FTY720 could lead to cell death. Bafilomycin A1, the inhibitor of autophagy, could enhance the cell viability in U266 cells treated with FTY720. NAC or Tiron, ROS scavenger, could decrease the FTY720 induced apoptosis and the expression of LC3B-II was reduced in combination of FTY720 with NAC or Tiron as compared with treatment with FTY720 only. It is concluded that FTY720 can induce U266 cell apoptosis and autophagy. ROS is the mediator that regulates both the apoptosis and autophagy in multiple myeloma cells.
1,2-Dihydroxybenzene-3,5-Disulfonic Acid Disodium Salt ; Apoptosis ; drug effects ; Autophagy ; drug effects ; Cell Line, Tumor ; Fingolimod Hydrochloride ; Humans ; Macrolides ; Microtubule-Associated Proteins ; metabolism ; Multiple Myeloma ; metabolism ; pathology ; Propylene Glycols ; pharmacology ; Reactive Oxygen Species ; metabolism ; Sphingosine ; analogs & derivatives ; pharmacology

Objective:To explore the clinical pathological characteristics and improve the recognition in the diagnosis and treatment of incidental (stage T1a -T1b)prostate cancer.Methods:Seven hundred and seventy-one patients who underwent TURP from May 2004 to September 2013 were analyzed retro-spectively.In our institution,TURP specimens should be totally submitted in an extensive sampling method.The tumor area was outlined by estimation of an experienced genitourinary pathologist and calcu-lated by the image analysis system software (Image J 1.47 h).The tumor area was then multiplied by the thickness of tissue.The total sum of all tumor volume was the estimated tumor volume.The clinical and pathological factors,follow-up results were obtained and we aimed to collect information about the period of watchful waiting (WW),PSA progression status,intervention status during the follow-up,the reason for intervention on WW and the type of intervention.Results:The average age of 771 patients was (71.3 ±5.9)years old,and the average BMI was (23.9 ±3.1)kg/m2 ,preoperative average tPSA was (4.4 ±2.8)μg/L.Eighty-six (11.2%)cases of incidental prostate cancer were detected.The patients in T1a group (77 cases,89.5%)had tumor volumes of (12.3 ±12.6)mm3 ,and the patients in T1b group had tumor volumes of (105.1 ±41.8)mm3 .The range of tumor volume was 0.4 -180.2 mm3 . The volume of all the 86 cases was less than 500 mm3 as the threshold of insignificant cancer.All the pa-tients were managed by WW.The mean follow-up time was 88.9 (27.9 -150.1)months.The Gleason score was <7 in 79 patients,and ≥7 in 7 patients.There was no significant difference in age,preopera-tive tPSA,preoperative PSAD,postoperative tPSA,prostate volume and TURP resection between T1a group and T1b group (P >0.05).Among 84 patients without follow-up losts,PSA progression occurred in 5 patients.One T1a patient underwent radical prostatectomy (RP)as an intervention,and 3 patients underwent hormone therapy.One patient in T1b group underwent radiotherapy for PSA progression and one was treated because of patient preference without evidence of disease progression.There were no pa-tients who died due to prostate cancer.Conclusion:Eighty-six (11.2%)cases of incidental prostate cancer were detected.The tumor volume of all the cases was insignificant cancer.The clinical outcomes of IPCa were satisfactory with the initial treatment of WW in the Chinese population.

Aged ; Arsenicals ; therapeutic use ; Female ; Humans ; Leukemia, Basophilic, Acute ; drug therapy ; Oxides ; therapeutic use

Gap junctions mediate direct communication between cells; however, toxicological cascade triggered by nonessential metals can abrogate cellular signaling mediated by gap junctions. Although cadmium (Cd) is known to induce apoptosis in organs and tissues, the mechanisms that underlie gap junction activity in Cd-induced apoptosis in BRL 3A rat liver cells has yet to be established. In this study, we showed that Cd treatment decreased the cell index (a measure of cellular electrical impedance) in BRL 3A cells. Mechanistically, we found that Cd exposure decreased expression of connexin 43 (Cx43), increased expression of p-Cx43 and elevated intracellular free Ca2+ concentration, corresponding to a decrease in gap junctional intercellular communication. Gap junction blockage pretreatment with 18β-glycyrrhizic acid (GA) promoted Cd-induced apoptosis, involving changes in expression of Bax, Bcl-2, caspase-3 and the mitochondrial transmembrane electrical potential (Δψm). Additionally, GA was found to enhance ERK and p38 activation during Cd-induced activation of mitogen-activated protein kinases, but had no significant effect on JNK activation. Our results indicated the apoptosis-related proteins and the ERK and p38 signaling pathways may participate in gap junction blockage promoting Cd-induced apoptosis in BRL 3A cells.
Animals ; Apoptosis/*drug effects ; Cadmium/*toxicity ; Calcium/metabolism ; Cell Communication/drug effects ; Connexin 43/genetics ; Enzyme Activation/drug effects ; Gap Junctions/*drug effects ; Gene Expression Regulation/drug effects ; Hepatocytes/cytology/*drug effects ; Rats ; Signal Transduction/drug effects

OBJECTIVETo study the expression changes of neuroglobin in rats with the model of diffuse traumatic brain injury and explore the relationship between the neuroglobin and neuron apoptosis in traumatic brain injury.

METHODSThe diffuse traumatic brain injury of rats was induced by the Marmarou's 'weight-drop' device. And the immunohistochemical technique was used to detect the expression changes of neuroglobin and neuron apoptosis in rat brain at different time points post-injury.

RESULTSThe expression of neuroglobin increased twice and reached peaks at 2 hours and 72 hours post-injury respectively. And the increased expression of neuroglobin from 30 minutes to 1 hour post-injury and from 48 hours to 72 hours post-injury accompanied with the decreased expression ratio of Bax to Bcl-2.

CONCLUSIONThe increased expression of neuroglobin in traumatic brain injury informed us that neuroglobin had anti-apoptosis action in post-injury neuron. It could protect the neuron from traumatic stress and secondary ischemia and hypoxia insults during ultra-early and acute stages.

Animals ; Apoptosis ; physiology ; Brain ; metabolism ; pathology ; Brain Injuries ; metabolism ; pathology ; Globins ; metabolism ; Male ; Nerve Tissue Proteins ; metabolism ; Neurons ; pathology ; Random Allocation ; Rats ; Rats, Sprague-Dawley

Objective To investigate the efficient method which can culture and induce embryonic stem cells to neurocyte in vitro. Methods Isolate the blastula of 3.5 d from BALB/c species mouse. Culture the cells from inner cell mass (ICM) which were isolated by mechanical method on the mouse embryonic fibroblaste cell (MEF) feeder layer or 0.1% gelatin coated dishes. The stem cells were identified by characterized morphology, alkaline phosphatase stain, differential potency in vivo and immunochemistry stain. The isolated cells were differentiated by serial induction method that mimicking the intrinsic developmental process of the neural system. Results The isolated cells were positive for alkaline phosphatatse and SSEA-1 (stage specific embryonic antigen 1). Moreover they were identified pluripotent by differentiation in vivo. Therefore the isolated cells presented the characters of ESCs. Then the isolated cells were able to differentiate into neurocytes in vitro. Conclusion Mouse embryonic stem cells isolation, culture and differentiation system has been established.

OBJECTIVETo study the expression of tau-related protein in spinal cord of Chinese patients with Alzheimer's disease.

METHODSGallays-Braak stain and immunohistochemical study for tau protein (AT8) were carried out in the spinal cord tissue (T2, T8, T10, L2 and S2 segments) of 3 Chinese patients with Alzheimer's disease. Seven age-matched cases without evidence of dementia or neurologic disease were used as controls.

RESULTSNeurofibrillary tangles were identified in the neurons of anterior horn in 2 Alzheimer's disease cases but none was observed in the controls. Tau-positive axons and astroglia were detected in all Alzheimer's disease cases. Tau immunoreactivity in spinal cord of the patients correlated with that in brain tissue.

CONCLUSIONThe expression of tau-related protein is demonstrated in the spinal cord of Alzheimer's disease patients suggesting that axonal transport defect may play a role in the pathogenesis of Alzheimer's disease.

Aged ; Alzheimer Disease ; metabolism ; pathology ; Axonal Transport ; Axons ; metabolism ; pathology ; Humans ; Male ; Neurofibrillary Tangles ; metabolism ; pathology ; Phosphorylation ; Spinal Cord ; metabolism ; pathology ; tau Proteins ; metabolism