1.Effect of human bone morphogenetic protein-7 gene transfection on proliferation of rabbit nucleus pulposus cells and synthesis of extracellular matrix
Jian SHI ; Xingang ZHAO ; Tiesheng HOU ; Xun TANG ; Yongqing XU
Chinese Journal of Tissue Engineering Research 2009;13(7):1389-1392
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.
3.Analysis of the cause of hemorrhage after MPCNL and its interventional treatment
Yongzhong HE ; Jian-He LIU ; Guo-Hua ZENG ; Jian YUAN ; Xun LI ; Zhao-Hui HE
Chinese Journal of Urology 2001;0(06):-
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.
4.Influence of Sedation and Analgesia on Stress Reaction of Post-Operation Infants with Congenital Heart Disease
jian-hui, ZHANG ; zhao-hui, CAI ; jie, HONG ; tu-xun, SI ; xiao-yuan, ZHAO
Journal of Applied Clinical Pediatrics 2006;0(23):-
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
5.Chemical constituents from barks of Nothopanax delavayi.
Qing YANG ; Jian ZHANG ; Sheng OUYANG ; Wen-Cai YE ; Shou-Xun ZHAO ; Zhi-Qi YIN
China Journal of Chinese Materia Medica 2014;39(10):1858-1862
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
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chemistry
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Drugs, Chinese Herbal
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chemistry
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Magnetic Resonance Spectroscopy
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Molecular Structure
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Plant Bark
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chemistry
6.Statistical methodologies used in genome-wide association studies
Feng CHEN ; Jian-Ling BAI ; Yang ZHAO ; Peng-Cheng XUN
Chinese Journal of Epidemiology 2011;32(4):400-404
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.
7.Chemical constituents from branch of Broussonetia papyrifera.
Jian-fei CHAO ; Zhi-qi YIN ; Wen-cai YE ; Shou-xun ZHAO
China Journal of Chinese Materia Medica 2006;31(13):1078-1080
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
8.Magnetic resonance imaging of eyes filled with silicone oil
Qing-Hua CHEN ; Xun YANG ; Fei YAN ; Zhen-Chang WANG ; Bo ZHAO ; Ya-Jian TONG ; Ying CUI ;
Ophthalmology in China 2006;0(05):-
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)
9.Experimental study of lentiviral vector-mediated bFGF gene transfection on rabbit BMSCs
Huai ZHANG ; Wu-Xun PENG ; Gang LIU ; Jian ZHANG ; Fei ZHANG ; Jian-Bo WANG ; Yin ZHAO ; Qing LI
Tianjin Medical Journal 2018;46(2):126-131
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.
10.Urinary S-phenylmercapturic acid variation in benzene exposed.
Yi-min LIU ; Hao CHEN ; Xu-dong LI ; Jian-xun HUANG ; Zhao-fa HUANG ; Min CAO
Chinese Journal of Industrial Hygiene and Occupational Diseases 2008;26(3):151-153
OBJECTIVETo observe the urinary S-phenylmercapturic acid (S-PMA) variation in the benzene dynamic exposed rat models and benzene exposed workers, and study the feasibility of use of urinary S-PMA as the biomarker in benzene exposed.
METHODSIn an animal model study, forty-eight adult Wistar rats were randomly divided into 4 groups: the control group, low-dose group, middle-dose group and high-dose group. The exposed groups were dynamically exposed for 28 days (4 periods) by benzene and the concentration was monitored. The urine was immediately collected after every exposure period and detected by the liquid chromatographic/mass spectrometry methods. In a cohort study, eighty benzene exposed workers in a ship-yard in Guangzhou were selected as the exposed subjects while forty healthy officers in the same shipyard who were not occupationally exposed to benzene were treated as the control. The urine was collected after work shift. The urinary S-PMA and the benzene in the workplace was treated as the rat model.
RESULTSIn the animal model study, the urinary S-PMA increased along with the environment benzene in every period and had significantly difference in the different exposed groups (P < 0.01 or P < 0.05), but did not change along with the exposed time course (P > 0.05). In the cohort study, the urinary S-PMA in the high-dose group [(27.2 +/- 7.9)microg/L] was significantly higher than the low-dose group [(13.6 +/- 3.4)microg/L] (P < 0.01). Otherwise, the background of urinary S-PMA was lower than 5microg/L in both workers and rat models.
CONCLUSIONThe urinary S-PMA can be proposed as a sensitive biomarker of occupational benzene exposure.
Acetylcysteine ; analogs & derivatives ; urine ; Adult ; Animals ; Benzene ; administration & dosage ; toxicity ; Environmental Exposure ; adverse effects ; Female ; Humans ; Male ; Middle Aged ; Rats ; Rats, Wistar ; Young Adult