3.Construction of autocatalytic caspase-3 driven by amplified human telomerase reverse transcriptase promoter and its enhanced efficacy of inducing apoptosis in human ovarian carcinoma
Yue SONG ; Keng SHEN ; Chun-Xia HE ;
Chinese Journal of Obstetrics and Gynecology 2000;0(09):-
Objective To construct recombinant adenoviral vector expressing autocatalysis caspase- 3 driven by human telomerase reverse transcriptase promoter amplified by two-step transcription amplification (hTERTp-TSTA),and investigate its antitumor effect in ovarian cancer iri vitro and in vivo.Methods Recombinant adenoviruses expressing autocatalytic caspase-3(rev-caspase-3)driven by hTERTp-TSTA were prepared,which were named as AdHTVP2G5-rev-casp3.AdHT-rev-casp3,Ad-rev-casp3 and AdHTVP2G5- EGEP,which express rev-easpase-3 driven by hTERTp,cytomegalovirus promoter(CMVp)and enhanced green fluorescent protein(EGFP),respectively,were used as controls.Western blot,cell counting kit (CCK-8),flow cytometry(FCM)and TdT-mediated dUTP-biotin nick end labeling(TUNEL)were used to detect the expression of p17,active subunit of caspase-3,and p85,and to measure cell survival rates, apoptotic rates and cell cycle distribution in ovarian cell line AO and normal human umbilical vein endothelial cell line HUVEC,following treatments of AdHTVP2G5-rev-casp3.subcutaneous tumor models and abdominally spread tumor models of human ovarian carcinoma using AO cells in BALB/e nude mice were established.Following treatments of AdHTVP2G5-rev-easp3,western blot was used to detect the expression of active caspase-3 in abdominally spread tumors and liver tissues,respectively,and the mouse survival rates and the volume of tumor nodules were measured,and the serum level of alanine transaminase (ALT)and aspartate transaminase(AST)were analyzed to monitor liver damages and HE staining was used to detect the histopathological changes of various organs.Results The levels of p17 expression in AdHTVP2G5-rev-casp3-treated AO cells were significantly higher than that in Ad-rev-casp3 or AdHT-rev- casp3 treated AO cells,while no expression was observed in AdHTVP2G5-rev-casp3-treated HUVEC.There was strong cell killing of AdHTVP2G5-rev-casp3 of hTERT positive AO cells,but not of the hTERT-negative HUVEC cells.Cell survival rate and apoptotic rate of AO cells treated with AdHTVP2G5-rev-casp3 were 17.8% and 40.2%,respectively,significantly different from that treated with AdHT-rev-casp3(75.2% and 16.1%)at the multiplicity of infection(MOI)of 70(P0.05) .Significant expressions of active caspase-3 were shown in AdHTVP2G5-rev-casp3-treated tumors,whereas no expression was shown in liver.In contrast,both tumors and liver tissues showed active caspase-3 expression following treatments of Ad-rev-casp3.AdHTVP2G5-rev-casp3 and Ad-rev-casp3 prolonged mouse survival[mean survival time of(259?14)d and(213?16)d],when compared with treatment with AdHT- rev-casp3[(177?12)d]and AdHTVP2G5-EGFP[(109?7)d;P
4.Changes of Surfactant Protein A and D in Bronchoalveolar Lavage Fluid in Human Fetal Lungs during Various Gestational Ages
yue, CHEN ; jiang, DU ; zhi-chun, FENG
Journal of Applied Clinical Pediatrics 2003;0(10):-
Objective To investigate the metabolic rules of surfactant protein A and D(SP- A,SP- D )in bronchoalveolar lavage fluid(BALF)in human fetal lungs during gestational ages. Methods BALF with 30 mL saline was performed on clinically collected human fetus with induction of labor by water- bag Their BALF was respectively retrieved [total retrieval rate(85. 6% ? 13 1)% ]for analysis of protein content. The BALF SP - A and SP - D from fetus of various gestational ages or newboms were detected by RPHA and ELISA. Results The total protein in BALF gradually increased since 10th week to newborn peak during lung development. And SP - A and SP D were respectively updated from(0.34 ?0.07 ) ,(0.05?0.01) ng/L to newborn climax[ (6 42 ? 0 36),(1.22 ? 0 13)ng/L] .Conclusions The protein in BALF gradually increases with fetal growth and lung development. SP-A and SP- D may reach prenatal climax and become the main indicator of newborn lung maturity.
5.Three new belamcandaquinones from Ardisia punctata
Chun LI ; Dangkun YUE ; Pengbin BU ; Youfu SUN
Acta Pharmaceutica Sinica 2006;41(9):830-834
Aim To study the chemical constituents of Ardisia punctata. Methods Compounds were separated with a combination of multi-chromatography. Their chemical structures were determined on the basis of spectral analysis and single crystal X-ray diffraction. Results Three compounds were isolated from chloroform extract of the roots of Ardisia punctata. Their structures were elucidated as 2-tridecyl-3-[ (2-tridecyl-3-acetoxy-4-methoxy-6-hydroxy) -phenyl ] -6-methoxy-1,4-benzoquinone ( 1 ), 2-tridecyl-3-[ ( 2 -tridecyl-4,6-dihydroxy ) -phenyl ] -6 -methoxy-1,4-benzoquinone ( 2 ) and 2 -tridecyl-3 - [ ( 2 -pentadecyl-4,6-dihydroxyl)-phenyl]-6-methoxy-1,4-benzoquinone (3). Conclusion The three compounds are new1,4-benzoquinone derivatives.
6.Monotherapy with levetiracetam for absence epilepsy in children
Yue HUANG ; Chun WU ; Jianjun WANG ; Liping ZHANG
Chinese Journal of General Practitioners 2011;10(12):873-875
Objective To assess the efficacy and safety of levetiracetam (LEV) monotherapy in children with absence epilepsy.Methods Fifty five children [ 34 males,21 females,aged (8.1 ± 4.8) y ] with typical absence seizures were treated with LEV ( n =25 ) or valproic acid ( VPA,n =30) from Jan 2009 to Jun 2010.Patients were followed up for 1y; the seizure frequency,electroencephalogram (EEG) and side effect were observed at 6th month and 12th month after the initiation of therapy.Results During the follow-up,none had aggravated symptoms of epilepsy.There were no significant differences in seizure frequency(t =0.76,P =0.450; t =0.67,P =0.504),abnormal EEG ( x2 =0.120,P =0.729 ; x2 =0.043,P =0.836 ) and seizure control ( x2 =0.051,P =0.821 ; x2 =0.078,P =0.780) after 6 and 12 months of treatment between two groups.No death or severe side effects were observed; 2 cases (8.0%) in LEV group had mild adverse effect( drowsiness and restlessness)and 2 cases (6.7% ) in VPA group had weight gain and nausea.Conclusion The results suggest that monotherapy with LEV is effective and safe for treatment of childhood absence epilepsy.
7.Two novel compounds from Ardisia punctata Lindl.
Chun LI ; Dangkun YUE ; Pengbin BU ; Youfu SUN
Acta Pharmaceutica Sinica 2007;42(9):959-963
To study the chemical constituents of Ardisia punctata,compounds were isolated with a combination of multi-chromatography.Their structures were determined on the basis of spectral analysis and comparison to those of the known compounds.A 1,4-benzoquinone derivative and a alkylphenol were isolated from the petroleum ether extract of the roots of Ardisia punctata.Their structures were elucidated as 2-tridecyl-3-[(2-tridecyl-4-acetoxy-6-methoxy)-phenoxyl]-6-methoxy-1,4-benzoquinone (1) and 2-methoxy-4-hydroxy-6-tridecyl-phenyl acetate (2).The two compounds are both new.