OBJECTIVETo investigate the effects of mifepristone on cell proliferation of human androgen-independent prostate carcinoma cell lines DU-145, PC-3 in vitro and the possible mechanisms involved.

METHODSThe A values of the prostate cancer cells DU-145 and PC-3 in each group with various concentrations (1, 10, 50, 100 micromol/L) of mifepristone at various time intervals (24-120 h) were detected with the colorimetric 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl tetrazolium bromide assay. The apoptosis rates of the DU-145 and PC-3 cells treated with 10 micromol/L of mifepristone for 24 h and 48 h were assessed by flow cytometry analysis technique. Immunohistochemical technique was used to determine the expression of bax, bcl-2 and vascular endothelial growth factor (VEGF) proteins after treatment with 10 micromol/L of mifepristone.

RESULTSThe A values of the cancer cells treated with 1 micromol/L of mifepristone were similar to that of controls, while those of the cells treated with 10 micromol/L, 50 micromol/L and 100 micromol/L of mifepristone were significantly different from that of controls (P < 0.01). Mifepristone markedly inhibited cell proliferation of prostate cancer cells DU-145 and PC-3 on a dose- and time-depending manner. The apoptosis rates of 10 micromol/L mifepristone for DU-145 cell line at 24 h, 48 h were respectively 15.3%, 30.4% with flow cytometry method and then PC-3 cell line were respectively 22.2%, 32.0%. Immunohistochemical technique showed the expression of bcl-2 and VEGF in the DU-145 and PC-3 cells treated with 10 micromol/L of mifepristone were significantly decreased, and the expression of bax was increased.

CONCLUSIONSMifepristone can induce apoptosis of androgen-independent prostate cancer cell lines DU-145 and PC-3 in vitro. The apoptosis effect is time-and-dose dependent. Mifepristone could initiate a cell death command via apoptotic pathways decreasing the expression of VEGF protein, downregulating the expression of bcl-2 protein and increasing the expression of bax protein.

Androgens ; metabolism ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Colorimetry ; Dose-Response Relationship, Drug ; Flow Cytometry ; Hormone Antagonists ; pharmacology ; Humans ; Male ; Mifepristone ; pharmacology ; Prostatic Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Time Factors ; Vascular Endothelial Growth Factor A ; metabolism ; bcl-2-Associated X Protein ; metabolism

Objective To survey knowledge of the local people's understanding of Brucellosis,and to explore the risk factors for brucellosis infection,and to determine the key issue of next comprehensive health promotion intervention.Methods Two counties,Yanggao and Guangling,which are old endemic areas with Brucellosis in history,and with epidemic rebounding in recent years,were selected.The survey was carried out by two stage stratified cluster sampling method.The questionnaires included respondents' demographic data(gender,age,education level,etc.),Brucellosis (hereinafter referred to as Brucellosis) knowledge of the investigation and behavior and attitude of people toward the measure for control of Brucellosis.Results A total of 5372 people were investigated in two counties of which 62.7％(3362/5372) of farmers.The investigated crowd had low culture level.The awareness of Brucellosis infection route of Yanggao and Guangling counties were 84.03％ (2379/2831) and 333％(847/2541 ).The average awareness of Brucellosis infection route was 18.60％(6001/32 260).In the investigation of knowledge on Brucellosis prevention of the two counties,29％ believed that it was necessary to wear gloves to process flow product,and 70％ of people answered do not know.For farmers on how to deal with dead animals,results showed that 79.1％(664/839) in Yanggao choose to sell dead animals to the market; 61.2％ (267/361) in Guangling choose to kill and bury,there were inappropriate treatment on handling of ill and dead animals in the two counties.Conclusions Spread of Brucellosis is caused mainly due to emphasis on the disease is not enough,and inappropriate handling of dead livestock.Measures like strengthening health education and behavioral intervention,increasing public awareness of the disease prevention and ability to change the incorrect way of life and cognitive concepts,can effectively reduce human infection and the spread of Brucellosis.

Objective To explore the clinical,radiological findings and pathogenic factor of spondylometaphyseal dysplasia.Methods Five cases were reported and the relevant documents were studied retrospectively.Plain X-ray film was performed in all patients.Results There were 4 male and 1 female,age ranged from 4 to 15 years with average of 9 years.The main features of 5 cases included delayed bone age,stature short,short trunk,waddling gait noted,scoliosis in 2 cases,kyphoscoliosis in 1 case,severe genu valgum in 2 cases.The main X-ray appearance of 5 cases is multiple irregularities of long bones metaphyses associated with platyspondylia,epiphysis is normal.Type Ⅰ SMD in 2 cases,Type Ⅱ SMD in 1 case,Type Ⅲ SMD in 2 cases.Conclusion When we meet children with delayed bone age and stature short,and muhiple irregularities of long bones metaphyses associated with platyspondylia were seen in X-ray plain film.we should think about spondylometaphyseal dysplasia.

0.05).Conclusions ATP-TCA could be used to individualize chemotherapy by selecting agents for particular patients of cervical cancer.The expression of GST-? and TS protein might be useful biomarkers to predict the resistance to DDP and 5-FU in patients with cervical cancer.

OBJECTIVERecent studies have found that the variation of G894T on the region of T786C and 7th exon promoted by endothelial nitric oxide synthase (eNOS) gene is associated with cardiovascular disease. This research explored possible correlations between eNOS gene polymorphisms and orthostatic intolerance (OI) in children through linkage disequilibrium analysis between eNOS genes T786C and G894T and OI.

METHODSPCR, Macrorestriction Map and other molecular biotechnology were used to determine the genotypes of eNOS/T786C and G894T in 60 OI probands and their parents. Correlation analysis and transmission disequilibrium test (TDT) between T786C, G894T and OI were performed.

RESULTSThere was linkage disequilibrium of eNOS/T786C and G894T gene polymorphisms in the occurrence of childhood OI (P<0.05).

CONCLUSIONSeNOS genes T786C and G894T may be associated with the pathogenesis of OI.

Adult ; Child ; Female ; Humans ; Linkage Disequilibrium ; Male ; Nitric Oxide Synthase Type III ; genetics ; Orthostatic Intolerance ; genetics ; Polymorphism, Genetic

OBJECTIVETo construct a vector expressing small interfering RNA (siRNA) against Rac1 gene and observe its effect on soft agar colony formation of SW480 cells in vitro.

METHODSOligos of 64 base pairs for hairpin RNA targeting Rac1 were chemically synthesized and annealed. The siRNA constructs for Rac1, produced by inserting the annealed oligos into the downstream of H1 promoter of linearized pSUPER, were confirmed by restriction digestion and DNA sequencing. The constructed Rac1-siRNA was transfected into SW480 cells and Western blotting was performed to assess the expression and interference efficiency of siRNAs against Rac1.The soft agar colony formation assay was used to study the effect of Rac1 gene silencing on SW480 cells.

RESULTSRestriction digestion and DNA sequencing showed that the siRNA targeting Rac1 gene was successfully constructed. The siRNA could effectively down-regulate the expression of Rac1 in SW480 cells. Soft agar colony formation assay showed that the colony number and diameter of SW480 cells was reduced after siRNA transfection.

CONCLUSIONA vector expressing hairpin RNA against Rac1 gene are successfully produced, which significantly reduces the colony numbers and size of SW480 cells in vitro, suggesting that Rac1 plays an important role in the growth of colorectal cancer in vitro.

Base Sequence ; Cell Line, Tumor ; Cell Proliferation ; Colonic Neoplasms ; pathology ; Down-Regulation ; Humans ; Molecular Sequence Data ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection ; rac1 GTP-Binding Protein ; genetics

OBJECTIVETo study the role of activation of Rac1 in colorectal cancer cell migration and invasion.

METHODSRac1 L61 plasmid and control plasmid were transfected into colorectal cancer cell line SW480 cells. Pull down assay by Western blotting was carried out to measure the amount of activited Rac1, and transwell permeable supports were used to assess the migration and invasion of SW480 cells with different activitivity of Rac1.

RESULTSThe transfection ratio of SW480 cells was more than 80%. Pull down assay showed that the activited Rac1 was significantly higher in the SW480 cells transfected with Rac1 L61 plasmid than that in the control, and the amount of migrating and invasing SW480 cells transfected with Rac1 L61 plasmid in the Transwell permeable supports were significantly more than those in controls (migrating cell numbers: 43 +/- 9 vs. 22 +/- 5, P < 0.01; invasing cell numbers: 73 +/- 13 vs. 38 +/- 1, P < 0.01).

CONCLUSIONThe activation of Rac1 plays an important role in the migration and invasion of colorectal cancer cells.

Cell Line, Tumor ; Cell Movement ; Colonic Neoplasms ; metabolism ; pathology ; Enzyme Activation ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Neoplasm Invasiveness ; Plasmids ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection ; rac1 GTP-Binding Protein ; genetics ; metabolism

OBJECTIVETo investigate the effects of X-ray exposure on the release of soluble tumor necrosis factor receptor-p75 (sTNFR-p75) in hepatocellular carcinoma HepG2 cells in vitro.

METHODSEnzyme-linked immunosorbent assay (ELISA) was used to examine the levels of sTNFR-p75 in the supernatants of HepG2 cells before and after X-ray exposure. The cell apoptosis was analyzed by flow cytometry and transmission electron microscope(TEM), and the morphological changes of the cells were examined under optical microscope and transmission electron microscope(TEM).

RESULTSX-ray exposure of the cells resulted in a strong increase of cell apoptosis (P<0.05) and sTNFR-p75 production in the cells as compared with the those before the exposure (P<0.01). Optical microscopy revealed apoptotic changes of HepG2 cell after the exposure, shown as cell shrinkage, spherical cell morphology, cytoplasmic and nuclear condensation. Apoptotic bodies were detected by TEM.

CONCLUSIONX-ray exposure induces HepG2 cells apoptosis by inhibiting the release of sTNFR-p75 into the supernatant.

Animals ; Apoptosis ; radiation effects ; Carcinoma, Hepatocellular ; pathology ; secretion ; Cell Line, Tumor ; Culture Media, Conditioned ; chemistry ; metabolism ; radiation effects ; Humans ; Liver Neoplasms ; pathology ; secretion ; Microscopy ; Receptors, Tumor Necrosis Factor, Type II ; biosynthesis ; chemistry ; secretion ; Solubility ; X-Rays

OBJECTIVEThe exercise rehabilitation in patient with chronic heart failure (CHF) is standard clinical practice, but it is rare using CardioPulmonary Exercise Testing (CPET) guide to prescribe exercise rehabilitation in China.

METHODSWe performed symptom limited maximal CPET in 10 patients with CHF, randomly divided into two groups: 5 patients as control without exercise and 5 exercise patients used Δ50%W intensity to exercise 30 min/d, 5 d/w, x12 w. Before and after 12 w rehabilitation, we evaluated functions.

RESULTSThere were no significant difference between two groups patients (P > 0.05). The exercise duration was increased from 8 min to 23 min after rehabilitation (P < 0.001); distance 6 minutes walking was increased from 394 m to 470 m (P < 0.05); score of Minnesota quality of life was decreased from 25 to 3 in exercise group (P < 0.01). However, there were nosignificant changes in control group (P>0.05) and their changes were smaller than those in exercise group (P < 0.01).

CONCLUSIONThe CPET guiding exercise rehabilitation is safe and effective for patients with CHF.

China ; Chronic Disease ; Exercise Test ; Exercise Therapy ; Heart Failure ; therapy ; Humans ; Quality of Life ; Walking

The study established a UPLC-MS/MS method that is used for simultaneous determination nine major bioactive compounds of Dachengqi Tang in rat plasma. Using Aglient C18 column (2.1 mm x 50 mm,1.7 microm) was chromatographed, using methanol-5 mmol x L(-1) ammonium formate mobile phase gradient, elution 0.3 mL x min(-1). In the plasma pre-treatment process, not only the method of methanol and acetonitrile protein precipitation was investigated, and different factors extraction solvent, the type of the scroll time, the number and the type of extraction solvent, the extraction volume of the extraction solution of liquid-liquid extraction is investigated. Finally, with ibuprofen as an internal standard, using ethyl acetate liquid-liquid extraction method pretreatment blood, N2 dry reconstituted supernatant after centrifugation UPLC-MS/MS analysis, in electrospray ionization (ESI) negative mode, using multiple reaction monitoring mode for testing. The linear range of emodin, rhein, aloe-emodin, chrysophanol, magnolol, honokiol, hesperidin and hesperitin is 0.33-660, 0.40-792, 0.41-827, 0.34-680, 0.45-907, 0.46-927, 0.43-867, 0.34-683, 0.39-787 microg x L(-1) respectively, good linear relationship; and extraction recovery were greater than 69.39%, days after the day of the RSD is less than 15%. This method can be used to study the rat gastric large bearing gas after Dachengqi Tang, the simultaneous determination of nine components in plasma for its pharmacokinetics and efficacy material base to provide a theoretical basis.
Animals ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Female ; Male ; Plasma ; chemistry ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; methods