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.

Background and Objective: Elevated expression of matrix metalloproteinases (MMPs) has been found in multiple carcinoma tissues.MMP-26 is highly expressed in prostate and breast cancer tissues,and promotes the invasion of human prostate cancer cells not only through the cleavage of fibronectin and type Ⅳ collagen but also by the activation of pro-MMP-9,a powerful gelatinase. This study was to present a comprehensive protein expression profile of MMP-26 in multiple human cancer tissues. Methods: The protein expression pattern of MMP-26 was examined using immunohistochemistry and multiple-tissue microarray. MMP-26 mRNA expression in coronary artery smooth muscle cells was detected by reverse transcription-polymerase chain reaction(RT-PCR). Results: The expression of MMP-26 in breast,colon,lung, brain, head and neck, prostate cancer, and melanoma tissues was significantly elevated when compared with parallel normal tissues (P＜0.05), while not significantly elevated in kidney cancer,ovarian cancer,and non-Hodgkin's lymphoma (P＞0.05).MMP-26 was also detected to express in gastric,rectal,thyroid, esophageal,and pancreatic cancers.MMP-26 protein was expressed in smooth muscle cells of the prostate and associated blood vessels. MMP-26 mRNA was also detected to express in human coronary artery smooth muscle cells. Conclusions: MMP-26 expression may be associated with multiple human carcinomas,and it may serve as a molecular marker for the early diagnosis of these carcinomas.MMP-26 may also contribute to smooth muscle function in the human prostate and cardiovascular system.

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

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 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

This study was aimed to develop and establish an efficient method for high through-put automatically extracting genomic DNA from EDTA-anticoagulated whole blood samples, and to utilize this method in routine rSSO HLA genotyping by luminex flow array assay, the genomic DNA was extracted automatically from 400 microl blood samples by using TECAN DNA workstation and 96-well plate with 2 ml volume per well. The yield and purity of each DNA sample was tested by UV-spectrophotometer, the integrity of these DNA samples were run electrophoresis on the agarose gel. Each DNA sample was subjected to PCR amplification and hybridization using One lambda rSSO HLA-A, -B and -DRB1 commercial kit, the fluorescent intensity for positive bead and negative bead hybridized with HLA-A, -B and -DRB1 PCR products were calculated and analyzed. The results showed that the mean yield and purity (A260/A280) of genomic DNA extracted from 400 microl whole blood samples were 3.217+/-0.715 microg and 1.710+/-0.103 respectively. The molecular weight was more than 15 kb in size and the fluorescent intensity for positive bead hybridized with HLA-A, -B and -DRB1 PCR products of each sample was >600 RFU, however, the fluorescent intensity for negative bead for each sample was <50 RFU. It is concluded that the highly qualified genomic DNA can be extracted automatically from blood samples of marrow-donors by using TECAN DNA workstation, and the extracted DNA samples are suitable for high through-put HLA genotyping by luminex flow array assay and other downstream transplant immunological and molecular biological experiments.
Biological Specimen Banks ; Bone Marrow ; DNA ; isolation & purification ; DNA Primers ; Genotype ; HLA Antigens ; genetics ; High-Throughput Screening Assays ; methods ; Humans ; Living Donors ; Nucleic Acid Hybridization ; methods ; Oligonucleotide Array Sequence Analysis ; methods

To establish the water dynamics model for drying process of Angelicae Sinensis Radix, the Weibull distribution model was applied to study the moisture ratio variation curves, and compared the drying rate and drying activation energy with the drying methods of temperature controllable air drying, infrared drying under different temperatures (50, 60, 70 degrees C). The Weibull distribution model could well describe the drying curves, for the moisture ratio vs. drying time profiled of the model showed high correlation (R2 = 0. 994-0. 999). The result proved that the drying process of Angelicae Sinensis Radix belonged to falling-rate drying period. For the drying process, the scale parameter (a) was related to the drying temperature, and decreased as the temperature increases. The shape parameter (β) for the same drying method, drying temperature had little impact on the shape parameter. The moisture diffusion coefficient increase along with temperature increasing from 0.425 x 10(-9) m2 x s(-1) to 2.260 x 10(-9) m2 x s(-1). The activation energy for moisture diffusion was 68.82, 29.60 kJ x mol(-1) by temperature controllable air drying and infrared drying, respectively. Therefore, the Weibull distribution model can be used to predict the moisture removal of Angelicae Sinensis Radix in the drying process, which is great significance for the drying process of prediction, control and process optimization. The results provide the technical basis for the use of modern drying technology for industrial drying of Angelicae Sinensis Radix.
Angelica sinensis ; chemistry ; Desiccation ; methods ; Models, Theoretical ; Water