BACKGROUND: Previous studies demonstrated that recombinant human bone morphogenetic pretein-7 (hBMP-7) can effectively promote the synthesis of extracellular matrix, repair of damaged disc matrix, restore of degenerative disc height. It is hoped that BMP-7 can be used to control and reverse the intervertebral disc degeneration. However, because of the short half-life of recombinant protein and low biological activity, it is difficult to maintain BMP-7 high concentrations on degenerative disc. Gene therapy can prevent these defects effectively.OBJECTIVE: To study the effect of hBMP-7 gene transfeotion on biological activity of primary cultured rabbit nucleus pulposus cells in vitro, to determine the feasibility of hBMP-7 gene which will on gene therapy research of intervertebral disc degeneration,and to provide basis for further study.DESIGN, TIME AND SETTING: A randomized and controlled observation was performed at the Institute of Cardiothoracic Surgery of Changhai Hospital from December 2005 to September 2006.MATERIALS: Six healthy New Zealand white rabbits of either gender, averaging 4 weeks old and weighing 500 g, were used in this study, and Ad-hBMP7 was constructed by the Cardiothoracic Surgery Institute of Changhai Hospital.METHODS: After rabbits sacrifice under aseptic condition, the nucleus pulposus was got. After digested with Pronase, type Ⅱcollagenase and type Ⅱ DNAase (4 hours, 37 "(2), the cells were seeded in the Petri dishes and were kept in the incubator. After 7 days and then twice a week, the media were refreshed. The nucleus pulposus cells were infected by adenovirus integrated with hBMP7 gene. The cells which were transfected by adenovirus vector for Lac-Z gene and which were not transfected were adopted as control.MAIN OUTCOME MEASURES: The expression of hBMP-7 was determined by RTopCR and Western blot. The effect of hBMP-7 on cell proliferation was surveyed by MTT. Furthermore, the effect of hBMP-7 gene on synthesis of proteoglycan and type Ⅱcollagen was detected by modified dimethylmethylene blue method and ELISA, respectively.RESULTS: Gene sequencing and PCR showed that hBMP-7 gene was inserted correctly and no mutation happened. The pdmary cultured rabbit nucleus pulposus cells were identical with those reported in literature. After Ad-hBMP7 transduct into the nucleus pulposus cells, high level of hBMP-7 expression was observed and lasted over 3 weeks. Also hBMP-7 gene can promote cell proliferation and synthesis of proteoglycan and type II collagen with significant difference compared with control groups (P < 0.05).CONCLUSION: hBMP-7 gene mediated by adenovirus can be the target gene for the further study on gene therapy of intervertebral disc degeneration.

Cerebral Hemorrhage ; diagnosis ; Cerebral Infarction ; diagnosis ; Diagnosis, Differential ; Humans ; Medicine, Chinese Traditional

Objective To analyze the cause of delayed hemorrhage after minimally invasive percu- taneous nephrolithotomy(MPCNL),and to summarize the experience in the interventional treatment of severe bleeding after MPCNL by superselective arteriolar embolization.Methods The clinical data of 3812 cases of MPCNL from June 1998 to July 2004 were reviewed.Of them,12 patients(11 men and 1 woman;mean age,45 years)who developed severe hemorrhage after MPCNL were identified.The cause of hemorrhage and the treatment results were analyzed.Results The rate of delayed hemorrhage after MPCNL was 0.31% (12/3812).The mean time to onset of severe bleeding was 10 d after MPCNL.Renal arteriography was per- formed in all 12 patients,showing 5 arteriovenous fistulas and 7 false aneurysms.Superselective arteriolar em- bolization for hemostasis was performed in all 12 cases.All these vascular abnormalities were successfully treated by superselective embolization.Follow-up showed that the hematuria disappeared and renal function recovered well.Conclusions Severe hemorrhage following MPCNL is a rare complication,the incidence of which is significantly lower than that of conventional PCNL.The cause is mainly the arteriolar injury of re- nal puncture passage.Superselective embolization provides effective control of bleeding and currently consti- tutes the treatment of choice based on our experience.

ObjectiveTo investigate influence of sedation and analgesia on stress reaction of post-cardiac surgery in infants with congenital heart disease.MethodsForty children with congenital heart disease were randomly divided into 2 groups after cardiac surgery.The analgesia group was given 0.5-2.0 ?g/(kg?h) fentanyl intravenous infusion in 20 children undergoing cardiovascular surgery.The control group was given 5-8 mg/(kg?dose) lbuprofen orally.Midaiolam 0.01-0.20 mg/(kg?h) was used in 2 groups for sedation by intravenous infusion or 0.05-0.10 mg/(kg?dose)by intravenous push intermittently.The effects and adverse effects of sedation and analgesia were observed on 2,8,24,48 h after surgery in each group.The levels of cortisol,growth hormone,insulin and blood glucose were measured,respectively.ResultsThere were significant differences in Ramsay,Comfort value on 2,8,24 h(Pa

Eleven compounds were isolated and purified from the barks extract of Nothopanax delavayi and their structures were identified as serratagenic acid-3-O-alpha-L-arabinopyranosyl-28-O-beta-D-glucopyranosyl ester (1), serratagenic acid-3-0-alpha-L-arabi-nopyranosyl-28-O-[alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl] ester (2), serratagenic acid (3), serratagenic acid-3-O-alpha-L-arabinopyranoside (4), serratagenic acid-beta-O-beta-(2', 4'-O-diacetyl) -D-xylopyranosyl-28-O-[alpha-L-rhamnopy-ranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->46)-beta-D-glucopyranosyl] ester (5), serratagenic acid-3-O-alpha-(4'-O-acetyl)-L-arabino pyrano-syl-28-0- [-alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl] ester(6), serratagenic acid-3-O-alpha-(2'-O-acetyl)-L-arabinopyranosyl-28-O-[-alpha-L-rhamnopyranosyl- (1-->4) -beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl] ester(7), serratagenic acid-3-0-beta-D-xylopyranosyl-28-O-[-alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl] ester (8), protocatechuic acid (9), ethyl caffeate (10) and caffeic anhydride (11) by physicochemical properties and spectroscopic data analysis. Among them, compounds 3-4 and 9-11 were firstly isolated from the genus Nothopanax, and compounds 5-8 were isolated from this plant for the first time.
Araliaceae ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Plant Bark ; chemistry

In lieu of large samples of cases and/or controls with hundreds of markers spreading throughout the human genome, researchers started to notice the dramatic increase of genome-wide association study (GWAS) for complex disorders, in the last 5 years. This paper highlights the statistical challenges in such huge-scale genetic studies, and introduces the analytical strategies and steps for handling GWAS data. Such issues as quality control of data, population stratification, methods available to data analysis and results presentation, replication, as well as the limitations of GWAS studies and the challenges presenting for statistics, are addressed.

OBJECTIVETo investigate the chemical constituents from the branch of Broussonetia papyrifera.

METHODColumn chromatographic methods were used to isolate the chemical constituents. ESI-MS and NMR methods were employed for their structural elucidation.

RESULTSix compounds were isolated and identified as (2S)-7, 3'-dihydroxy-4'-methoxyflavan (1), ergosterol peroxide (2), D-galacitol (3), sulfuretin (4), liriodendrin (5), graveolone (6), respectively.

CONCLUSIONCompounds 1-6 were isolated from the plant for the first time.

Benzofurans ; chemistry ; isolation & purification ; Broussonetia ; chemistry ; Ergosterol ; analogs & derivatives ; chemistry ; isolation & purification ; Flavonoids ; chemistry ; isolation & purification ; Furans ; chemistry ; isolation & purification ; Glucosides ; chemistry ; isolation & purification ; Plant Stems ; chemistry ; Plants, Medicinal ; chemistry

Objective To evaluate values of magnetic resonance imaging(MRI)in eyes filled with silicone oil.Design Prospective cases series.Participants 40 eyes of 40 patients were filled with silicone oil after ocular injury.Methods MRI was performed in the 40 patients,including axial FSE T_1WI,T_2WI,coronal T_2WI with fat saturation,oblique sagittal T_1WI and axial T_2FLAIR.MRI findings,in- cluding morpbous,signal and complications were analyzed.Oculi axes were measured.Main Outcome Measures Morphous,signal, complications and oculi axes of the eyes filled with silicone oil.Results Affected oculi axis was 2.18cm?0.21cm,normal oculi axis was 2.48cm?0.16cm.The silicone oil in eyes demonstrated isointense signal or slightly hyperintense signal on T_1WI and T_2WI,hypointense signal after fat saturation.Hydrops was found in vitreous cavity in 33 patients,including simple hydrops in 17 patients and complicated other abnormality in 16 patients.Choroidal detachment was found in 11 patients,complicating vitreous hydrops in 5 patients and lo- calized bulge of eyeball wall.Retinal detachments were found in 4 patients,of whom 3 patients complicated with vitreous hydrops.Per- fluorocarbon liquid residual in vitreous cavity,foreign body in anterior chamber,localized thickness of the wall of the globe and meagre- mean of silicone oil in vitreous cavity were found respectively in one patient complicating vitreous hydrops.Except for eye changes, fracture of orbital wall and foreign body in orbit were found in one patient.Conclusions MRI can display the changes of eyes filled with silicone oil,and measure oculi axes biologically and accurately offering important clinical application value.(Ophthalmol CHN,2007,16: 312-315)

Objective To observe the effect of lentiviral vector-mediated basic fibroblast growth factor (bFGF) gene transfection on the biological characteristics of rabbit bone marrow stromal cells(BMSCs)under in vitro culture conditions. Methods BMSCs were obtained by density gradient centrifugation and adherence screening. The bFGF gene was transfected into BMSCs by lentiviral vector and divided into bFGF transfection group,empty virus group and untransfected group according to the transfection conditions.After transfection,the morphology,expressions of bFGF mRNA and protein, cell proliferation,cell cycle and alkaline phosphatase(ALP)activity were observed in three groups of cells. Results High density BMSCs were successfully obtained by density gradient centrifugation and adherence screening.After transfection of BMSCs with bFGF gene, the cell morphology showed no significant changes, while the expressions of bFGF mRNA and protein were significantly increased, the cell proliferation curve shifted upward, the proportion of proliferating cells increased,and the activity of ALP was significantly enhanced.There were significant differences between three groups(P<0.05).Conclusion The rabbit bFGF gene is successfully introduced into the BMSCs cultured in vitro by lentiviral vector, and the target gene is stably expressed.The expression of bFGF can promote the proliferation and osteogenic differentiation of BMSCs.

The hydration reaction by microbial method is the crisis of the procedure of acrylamide production from acrylonitrile. This research studied the enzyme catalytic kinetics and de-active kinetics of nitrile hydratase in the type of free cell. Firstly, the effects of the concentration of cells, the temperature, pH value, the concentration of acrylonitrile and the concentration of acrylamide on the activity of nitrile hydratase was investigated. The result is that the temperature and the concentration of acrylamide are the most important among these factors. The activity of the nitrile hydratase was 5659 u/mL (broth) at 28 degrees C; the counterpart was only 663 u/mL (broth) at 5 degrees C. And the activity of NHase in solution of 45% acrylamide was just about half of that in solution of 5% acrylamide. After study on the relation of temperature and the reaction speed, It was found that the activation energy of the hydration of NHase was 65.57 kJ.mol-1. This paper studied the effects of concentration of cells, temperature, pH value, concentrations of acrylonitrile and acrylamide on the deactivation of Nhase, as well as the related enzyme de-active kinetics. The result also showed that the temperature and the concentration of acrylamide are the most important among these factors. In solution of 35% acrylamide, the residual activity was about 0% of the original value after 55 h; but in solution of 10% acrylamide, after the same period of time, the residual activity was 50% of the original one. It was also found that the concentration of acrylonitrile had little effect on the stability of NHase. The coefficient of deactivation at 28 degrees C was 21.77 times of the one at 5 degrees C. Correlating the temperature and the coefficient of deactivation, the activation energy of the de-active reaction was found to be 92.28 kJ.mol-1.
Acrylamide ; metabolism ; Acrylonitrile ; metabolism ; Catalysis ; Hydro-Lyases ; metabolism ; Hydrogen-Ion Concentration ; Kinetics ; Rhodococcus ; enzymology ; Temperature