A 2,4 -dichlorophenol degrading Pseudomonas strain GI241-1 was isolated from a soil sample. The dienelactone hydrolase gene, designated as dcpD which encodes dienelactone hydrolase involved in transforming cis-2-chloro-dienelactone into 2-chloromaleylacetic acid, was cloned from this bacterium strain. The gene cloning strategy was to construct genomic library after location of its neighbouring gene by Southem blot and to screen the aim transformant by dot blotting. Sequencing results showed that length of dcpD is 702bp. The sequence of dcpD and the deduced amino acid are different from the relative sequences registered in the GenBank.

Zwittermicin A was purified by ion exchange resin and HPLC from supernatants of Bacillus thuringiensis.subsp.kurstaki strain D1-23 cultivation.2.89mg pure Zwittermicin A was acquired,proved by HPLC-MS.Results show that the optimized wash concentration of NH_ 4 H_ 2 PO_ 4 is 5mmol/L at first step.Next step CH_ 3 COONH_ 4 concentration is 30 mmol/L,the gradient pH is 8.0～9.5.Totally 93% Zwittermicin A can be reserved with ion exchange resin.The temperature and pH stability experiments show the half life of Zwittermicin A is 48.22 minutes in 100℃,and it is more stable in lower pH in pH 2.0～12.0.

Objective To observe the relationship between apoptosis of K562 cell induced by iron-deprivation and activation of Caspase-3.Methods K562 cells were treated with desferrioxamine(DFO) in different dosages were collected at different time points.K562 cells were labelled with Annexin V/PI,and then the rate of apoptosis was measured by flow cytometry;The activation of Caspase-3 were detected by colorimetric method with pAN labelled substrate;The active protein of Caspase-3 were analyzed by Western blot.Results When K562 cells were treated with different concentrations of DFO,the apoptosis rate and the activity of Caspase-3 increases gradually.When K562 cells were incubated with DFO(50 ?mol/L and 100 ?mol/L) 24 h later,the enzymatic activity of Caspase-3 increases dramatically more than that of control group,and the difference was significantly(P0.05).All those effect above can be counteracted by equal mole concentration of FeCl_3.Conclusion Iron-deprivation maybe induce the apoptosis of K562 cell by chelating intracellular iron and activing Caspase-3.

OBJECTIVETo investigate the role of c-Jun and c-Fos as transcriptional factors in regulation of dentin sialophosphoprotein (DSPP) gene by a promoter-luciferase reporter gene construct in odontoblast cell line MDPC-23.

METHODSEndogenous c-Jun or c-Fos protein was determined by immunocytochemistry. The role of c-Jun or c-Fos in transcription of DSPP was investigated in co-transfection experiments using promoter-luciferase reporter gene construct containing the sequence between -791 bp and +54 bp of mouse DSPP gene.

RESULTSc-Jun and c-Fos was expressed by MDPC-23 cells, and located in the nucleus of MDPC-23 cells. Overexpression of c-Jun or c-Fos significantly inhibited luciferase activity of DSPP promoter.

CONCLUSIONThese findings suggest c-Jun and c-Fos down-regulated the transcription of DSPP gene as a transcriptional factor in odontoblast.

Animals ; Cell Line ; Extracellular Matrix Proteins ; Gene Expression Regulation ; Mice ; Odontoblasts ; Phosphoproteins ; Promoter Regions, Genetic ; Sialoglycoproteins ; Transfection

2,4-Dichlorophenol is toxic and biorefratory organic pollutant. A 2,4-dichlorophenol degrading bacterial strain GT241-1, identified as Pseudomonas sp., was isolated from soil samples which was collected from drainage area of several 2,4-dichlorophenol producing factories. Strain GT241-1 had strong 2,4-dichlorophenol degrading ability, it could decompose 91% 2, 4-dichlorophenol of 90 mg/L within 48 hours at 25 - 30 degrees C, and could utilize 2,4-dichlorophenol, 2,4-dichlorophenoxyacetic acid (2,4-D), benzoate and catechol as sole carbon and energy source. Southern blot showed that 2,4-dichlorophenol hydroxylase gene (dcpA) of strain GT241-1 locates on the about 10kb EcoR I/Xba I fragment. This fragment was recovered, linked to the vecter pUC19 and transformed into the E. coli DH5alpha. A aim transformant, Z539, was obtained by dot blotting from about 1200 transformants. PCR and the sequencing results shew that the whole dcpA gene is contained within the 10kb EcoR I /Xba I fragment of pZ539. This fragment was shortened to about 2.4kb by HindmIII. The shorted fragment was subcloned to vecter pRSET-B to get a transformant BS1-12. The subcloned fragment was sequenced. Sequencing results showed that the whole length of the subcloned fragment containing dcpA is 2389bp and the nucleotide span of coding region is from number 276 to number 2072 (1797 bp), with ATG and TAA as start and stop codon respectively. The sequence analysis of dcpA and the deduced amino acid encoded by dcpA showed that they are different from the relative sequences registered in the GenBank. The subcloned fragment carry the promoter of dcpA, this can deduce from the fact that the upflow length of dcpA coding region is 275bp, and further confirmed by the 2,4-dichlorophenol hydroxylase activity measurement results. The 2,4-dichlorophenol hydroxylase activity of transformant Z539 and BS1-12 were detected, the results showed these transformants have 2,4-dichlorophenol hydroxylase activity. By comparison, the activity of these transformants were lower than that of the strain GT241-1.
Amino Acid Sequence ; Bacterial Proteins ; genetics ; metabolism ; Biodegradation, Environmental ; Chlorophenols ; metabolism ; Cloning, Molecular ; Environmental Pollutants ; metabolism ; Mixed Function Oxygenases ; genetics ; metabolism ; Molecular Sequence Data ; Pseudomonas ; enzymology ; genetics ; isolation & purification ; Soil Microbiology

Objective To evaluate the efficacy and safety of acute ureteroscopy for the treatment of ureterie stones during middle and late pregnancy.Methods From June 1998 to March 2005,17 pregnant women(mean age,27 years;age range,21-35 years)with ureteric stones were treated by ureteroscopy when the fetus was at 20-36 weeks of gestation(mean,29 weeks).All the cases presented with urgent symptoms such as recurrent renal colic(11 cases),fever(4)or acute obstructive anuria(2).Among 17 cases,the stones(between 6 mm?7 mm and 13 mm?21 mm)were located in the upper(8 cases),middle(5)or lower ureter(4);and on the left side(5 eases),on the right(10)and on both(2)of the lower ureter. Mild hydronephrosis were observed in 6 cases and moderate hydronephrosis in 11,Of the 17 cases,14 under- went ureteroscopic pneumatic lithotripsy;in 1 case the calculi were pushed to the renal pelvis;and 2 cases were treated by Double-J catheter drainage.Results All the urgent symptoms in 17 cases were relieved after treatment.The stone-free rate of initial treatment was 82.4%(14 of 17).Three cases with residual stones were treated by Douhle-J catheters,which were replaced every 3 months until the calculi were re- moved.No abortion,premature delivery or complications such as ureter perforation occurred.Mild renal colic occurred in 1 case after insertion of Douhle-J catheter,and it was relieved 3d later;gross hematuria occurred in l case and disappeared 6 d later without treatment.All 17 patients had normal delivery and gave birth to healthy children.Conclusions Ureteroscopy is a safe and reliable method for the treatment of ureteric calculi during middle and late pregnancy.

OBJECTIVETo characterize the role of Smads proteins in alpha 2 (I) collagen (COL1A2) gene expression induced by bone morphogenetic protein-2 (BMP-2) in odontoblast cell line MDPC-23.

METHODSEndogenous Smad protein expression was determined by immunocytochemistry. Smads function and their role in COL1A2 gene expression were investigated in cotransfection experiments using promoter-luciferase reporter gene construct.

RESULTSMDPC-23 cells expressed Smad1, Smad5 and Smad6. BMP-2 promoted the activation of COL1A2 promoter reporter construct. Transient overexpression of Smad1 or Smad5 was enhanced, while overexpression of Smad6 inhibited BMP-2-induced COL1A2 promoter activity. BMP-2 inducibility could be blocked by overexpression of Smad1 or Smad5 dominant negative mutant.

CONCLUSIONSSmad signaling is functioning and appears to be involved in BMP-2-induced COL1A2 collagen transcription in MDPC-23. Smad signaling may play an important role in odontoblast differentiation and dentin extracellular matrix formation mediated by BMP-2.

Animals ; Bone Morphogenetic Protein 2 ; Bone Morphogenetic Proteins ; genetics ; Cell Line ; Collagen ; genetics ; Collagen Type I ; Mice ; Odontoblasts ; cytology ; metabolism ; Smad Proteins ; physiology ; Transforming Growth Factor beta ; genetics

Carnosol has been proved to have anti-breast cancer effect in previous research. But its ER subtype's specific regulation and mediation mechanisms remain unclear. The aim of this study is to observe the effect of carnosol on cell proliferation and its estrogen receptor α and β's specific regulation and mediation mechanisms with ER positive breast cancer T47D cell. With estrogen receptor α and β antagonists MPP and PHTPP as tools, the MTT cell proliferation assay was performed to observe the effect of carnosol on T47D cell proliferation. The changes in the T47D cell proliferation cycle were detected by flow cytometry. The effect of carnosol on ERα and ERβ expressions of T47D cells was measured by Western blot. The findings showed that 1 x 10(-5)-1 x 10(-7) mol x L(-1) carnosol could significantly inhibit the T47D cell proliferation, which could be enhanced by MPP or weakened by PHTPP. Meanwhile, 1 x 10(-5) mol x L(-1) or 1 x 10(-6) mol x L(-1) carnosol could significantly increase ERα and ERβ expressions of T47D cells, and remarkably increase ERα/ERβ ratio. The results showed that carnosol showed the inhibitory effect on the proliferation of ER positive breast cancer cells through target cell ER, especially ERβ pathway. In the meantime, carnosol could regulate expressions and proportions of target cell ER subtype ERα and ERβ.
Blotting, Western ; Breast Neoplasms ; metabolism ; pathology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Diterpenes, Abietane ; chemistry ; pharmacology ; Dose-Response Relationship, Drug ; Estrogen Receptor Modulators ; pharmacology ; Estrogen Receptor alpha ; antagonists & inhibitors ; metabolism ; Estrogen Receptor beta ; antagonists & inhibitors ; metabolism ; Female ; Flow Cytometry ; Humans ; Molecular Structure ; Pyrazoles ; pharmacology ; Pyrimidines ; pharmacology

BACKGROUNDVascular control and tissue dissection are crucial steps in successful laparoscopic surgery. Recently, a new commercially available vessel sealing technology, the LigaSure vessel sealing system (Valleylab, Boulder, USA), has been introduced. The aim of the present study was to evaluate the benefits of the LigaSure in laparoscopic nephrectomy.

METHODSFrom January 2005 to March 2010, 170 laparoscopic nephrectomies were performed with the LigaSure vessel sealing system, including simple and radical nephrectomy and nephroureterectomy. In a retrospective study, the laparoscopic operating time, estimated intraoperative blood loss, duration of postoperative drainage, total amount of postoperative drainage, as well as postoperative hospital stay, were recorded and studied.

RESULTSAll 170 laparoscopic nephrectomies using LigaSure were accomplished successfully without conversion to open surgery. There was no severe vascular complication or other serious complications. The mean laparoscopic operating time was 124.2 minutes (range, 14 - 230 minutes); mean blood loss was 148.6 ml (range, 20 - 540 ml); mean time for postoperative drainage was 3.1 days (range, 1 - 7 days); mean amount of postoperative drainage was 206.5 ml (range, 27 - 435 ml) and mean postoperative hospital stay was 6.9 days (range, 3 - 18 days).

CONCLUSIONSLaparoscopic nephrectomy using LigaSure appears technically feasible and easy, and produces satisfactory results. The LigaSure provides a safe and fast way to seal vessels and tissue bundles during nephrectomy.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Laparoscopy ; methods ; Male ; Middle Aged ; Nephrectomy ; methods ; Retrospective Studies ; Young Adult

Objective To study the dosimetry distribution of 125Ⅰ seed chains with different radians in different curvatures of bile ducts. Methods The outlines were drawn on the papers, which are the seed chain models with different radians. Radians formula (radian length=2πr × angle/360) was used to calculate the corresponding 0°, 30°, 60°, 90°, 120°, 150° and 180° models with a radian length at 45 mm, for the total length of seed chain model was 45 mm, and the seeds, had no interval or linear arrangements. The image was transmitted to the Brachytherapy planning system for seeds implantation( TPS) to simulate the seed chains with different radians. Using TPS to delineate the tumor target area, of which the activity was set as 1. 85 × 107 Bq, and the prescription dose was 60 Gy. It was prescribed to simulate the bile duct ( diameter at 8 mm) . TPS were used to calculate the D90 and V100 of the simulated bile duct with the diameter at 8 mm, and explore dosimetry of the points at the centripetal and centrifugal sides with 5 mm vertical distance which from two endpoints and center of seed chains with different radians. Results When the radian of seed chain was 30°, the D90and the V100 were the highest (the D90 was 132 Gy; the V100 was 100%). While the radian was 60°, the D90 and the V100 were the lowest (the D90 was 45 Gy, the V100 was 68%). As the radian was 30°, the highest dose was in the center ( dose in the centripetal side was 165 Gy, and centrifugal side dose was 142 Gy) . The center has the lowest dose as the radian up to 180°(dose in the centripetal side was 90 Gy, and centrifugal side dose was 50 Gy) . Among all radians, dose in the centripetal side was always higher than centrifugal side in the center. Between two endpoints, dose in the centrifugal side was higher than centripetal. Conclusions Distribution of seed chain dosage also changed along with the change of radian. When the radian of seed chain was 30°, the D90 and the V100 were the highest. The centripetal dose was higher than that of the centrifugal side.