1.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
2.STUDIES ON COMPOSITION,ANTIOXIDATION AND HYPOGLYCEMIC EFFECTS OF POLYSACCHARIDES FROM GREEN TEA, OOLONG TEA AND BLACK TEA NI
De-Jiang NI ; Yu-Qiong CHEN ; Bi-Jun XIE ; Chun-He SONG ; Fang-Ting YUAN ;
Acta Nutrimenta Sinica 1956;0(01):-
Objective: To compare tea polysaccharides(TPS) characteristics and their role in scavenging free radicals and reducing blood glucose(BG) in diabetic mice(DM). Methods: TPS was extracted from green,Oolong and black tea which were made from the same fresh leaves from Hubei,Fujian and Yunnan. Then the recovery rate of TPS, contents of neutral sugar, uronic acid and protein were analysed, and scavenging rate of -2Oand 稯H in vitro and hypoglycemic effect were also determined. Results: 1. The yield and contents of neutral sugar, uronic acid and protein of green tea TPS were the highest, and those of black tea TPS were the lowest. Oolong tea TPS acted the best in scavenging-2O and 稯H . 2. The hypoglycemic effect of TPS from Hubei tea was the best . The effect of TPS extracted from semi-fermented Oolong tea and fermented black tea was better than that of non-fermented green tea. 3. There were obvious differences in yield, free radical scavenging rate and effect of reducing BG among TPS extracted from tea in different regions. TPS extracted from Fujian tea had the best effect in reducing BG,but that from Yunnan tea had not. Conclusion: There was remarkable effect of region and process on physico-chemical characteristics,effect of scavenging radical and reducing blood sugar TSP.
3.Effects of the Culture Method on the Construction of Dermal Substitutes in vitro
Yan-Na HE ; Chun-Mei DING ; Hua JIANG ; Yan ZHOU ; Wen-Song TAN ;
China Biotechnology 2006;0(05):-
Culture environment is the key factor in the construction of dermal skin.It was investigated that the effects of the culture methods,including the static culture and spinner flask culture,and stir speeds on the cells proliferation,metabolism and distribution within collagenchitosan sponges.A higher cell density and specific growth rate was obtained with spinner flask culture versus static culture,especially,the 80 r/min spinner flask culture.The cell distribution in dermal substitutes from stirred culture system was more uniform than static culture,as well as that with increase of stir speeds in spinner flask.In summary,the spinner flasks culture with proper stir speed shows promise for the construction of dermal substitutes in vitro.
4.The effect of estradiol and testosterone on the apoptosis of lacrimal gland cell induced by H2O2
Xiang-yin, SHA ; Chun-yun, LUO ; Li, SONG ; Fei-hong, FAN ; Xiao-song, HE ; Dao-bing, DU
Chinese Journal of Experimental Ophthalmology 2011;29(12):1082-1086
Background The sex hormones plays an important role in the incidence of dry eye,especially for the regulation of function.However,the effects of sex hormones on lacrimal gland epithelial cells are below understand.Objective This study was to investgate the effects of estradiol and testosterone on the apoptosis of lacrimal gland cells induced by H2O2.Methods The lacrimal gland tissue was obtained from 2- or 3-month-old clean male New Zealand rabbits and the lacrimal gland epithelial cells were cultured in vitro using esplant culture method.The cells were identified by pan cytokeratin antibodies with immunocytochemistry.lacrimal gland epithelial cells were incubated in the 96 well plate at the density of 5 × l04 cells/ml for 44 hours.Estradiol or testosterone with the concentrations of 1 × 10-5,1 × 10-6,1 × 10-7,1 × 10-8 mol/L were added into the medium for 24 hours respectively and 1× 10-4 mol/L H2O2 treated the cells for 1 hour to induce the apoptosis in experimental groups.The cells treated by only 1 × 10-4 mol/L H2O2 were used as apoptotic control group,and the cells cultured by regular method were used as blank control group.The cell viability in different groups was detected using MTT at 570 nm ( A570 ),and the apoptotic rates of the cells were assayed using Annexin V/PI double staining.This use and maintain of experimental animals followed the Regulation for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission.Results The cultured cells showed the irregular polygon in shape,and about 80% cells was positive response for cytokeratin.MTT assay showed that the lower A570 values were detected in the H2O2-induced group,various concentrations of estradiol or testosterone groups compared with blank control group (P<0.01 ).The A570 values in 1 × 10-5,1 × 10-6,1 × 10-7 mol/L estradiol groups or 1 × 10-6 mol/L testosterone group were significantly higher than ones of H2 O2-induced group (P<0.01 ).Compared with corresponding concentrations of testosterone groups,the A570values in various concentrations of estradiol groups were elevated( P<0.01 ).The apoptosis rates at the early and later phase were significantly declined in both estradiol group and testosterone group in comparison with H2 O2-induced group (P < 0.01,P< 0.05 ),and those in estradiol group were lower than the testosterone group( P<0.01,P<0.05 ).Conclusions Estradiol and testosterone suppress the apoptosis of lacrimal gland cells induced by H2O2,and the stronger effect is found in estrogen.The inhibition of estrogen on lacrimal gland cell apoptosis show a dose-dependent manner to some extent.
5.Application of Susceptible-Infected-Recovered model in dealing with an outbreak of acute hemorrhagic conjunctivitis on one school campus
Tian-Mu CHEN ; Ru-Chun LIU ; Qi-Qi WANG ; Song-Lin ZHU ; Ai-Chun TAN ; Qiong HE ; Xin LIU ; Guo-Qing HU
Chinese Journal of Epidemiology 2011;32(8):830-833
To simulate intervention measures in controlling an outbreak of acute hemorrhagic conjunctivitis on one school campus by using the Susceptible-Infected-Recovered (SIR) model, to provide evidence for preparedness and response to the epidemic. Classical SIR model was used to model the epidemic. Malthusian exponential decline method was employed to estimate the infective coefficient β for interventions. The initial value of parameters was determined based on empirical data. The modeling was implemented using Matlab 7.1 software. Without interventions, the outbreak was expected to experience three phrases: (1)early stage (the first 5 days) in which the epidemic developed slowly and could be intervened easily; (2) rapid growing stage (6-15 days) in which the number of infected cases increased quickly and the epidemic could not be well controlled;and (3) medium and late stage (16 days and later) in which more than 90% of the susceptible persons were infected but the intervention measures failed to prevent the epidemic. With the implementation of interventions, the epidemic was predicted to be controlled in the early stage, under the SIR model. The simulation based on the SIR model kept an acceptable consistency with the actual development of epidemic after the implementation of intervention measures. The SIR model seemed effective in modeling interventions to the epidemic of acute hemorrhagic conjunctivitis in the schools.
6.Pulmonary Embolism following Operation for Femur Fracture under General Anesthesia: A case report.
Seong Hoon KO ; Jeong Ja SONG ; Chun Won YOO ; Sang Kyi LEE ; He Sun SONG
Korean Journal of Anesthesiology 1996;31(4):519-524
Pulmonary embolism is a common and lethal, which accounts for substantial morbidity and mortality. Clinical manifestations of pulmonary embolism are nonspecific during general anesthesia. A 60 years old female received elective operation for left femur fracture under general anesthesia. At the end of operation, she suddenly became hypotensive and developed cyanosis. Immediate cardiopulmonary resuscitation(CPR) was performed without definitive diagnosis. Pulmonary embolism was suspected by clinical signs and echocardiography. So, patient was transferred to intensive care unit and with intensive care and aggressive treatment, patient's vital signs and ventilatory status were progressively improved. However, the endotracheal tube was accidentally extubated by the patient at the second postoperative day, and then cardiac arrest was developed and the patient expired. The primary goal of therapy for pulmonary embolization is to prevent reembolization. In the pulmonary thromboembolization, early diagnosis and intensive care improve outcome.
Anesthesia, General*
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Cyanosis
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Diagnosis
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Early Diagnosis
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Echocardiography
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Embolism
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Female
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Femur*
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Heart Arrest
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Humans
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Critical Care
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Intensive Care Units
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Middle Aged
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Mortality
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Pulmonary Embolism*
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Vital Signs
7.Preparation of in situ gel systems for the oral delivery of ibuprofen and its pharmacokinetics study in beagle dogs.
Rui-ling WU ; Chun-shun ZHAO ; Jing-wen XIE ; Shao-ling YI ; Hong-tao SONG ; Zhong-gui HE
Acta Pharmaceutica Sinica 2008;43(9):956-962
The in situ gel systems can form gel in situ after administration to achieve sustained release, thus provides a promising strategy for drug delivery systems. The aim of this study was to design and prepare in situ gel systems for the oral delivery of ibuprofen (IBU-ISG) and study its pharmacokinetics in Beagle dogs. The characteristics of the basic material of gellan gum (Kelcogel, Kel) and sodium alginate (Manugel, M) were studied through investigating the complex viscosity of the Kel or M solution with or without different concentrations of calcium ion or sodium citrate to ascertain the amount range of the excipients. The measurement of complex viscosity of the solution (0. 5% Kel and 1% M) with different concentrations of sodium citrate and calcium ion was carried out to select the suitable proportion of calcium ion and sodium citrate. The formulation of binary IBU-ISG was optimized by monitoring the complex viscosity before gelling in vitro release property. The optimized formulation contains 1.0% sodium alginate, 0.5% gellan gum, 0. 21% sodium citrate and 0.056% calcium chloride. A single oral dose of IBU-ISG and reference formulation (IBU suspension) were given to each of the 6 healthy Beagle dogs, ibuprofen in plasma at different sampling times was determined by RP-HPLC. The pharmacokinetics parameters in 6 Beagle dogs were calculated. The Tmax of IBU-ISG and reference formulation were (1.8 +/- 0.6) and (0.4 +/- 0. 1) h. The Cmax values were (29.2 +/- 7.6) and (37.8 +/- 2.2) microg x mL(-1). The T(1/2) were (2.3 +/- 0.5) and (2.0 +/- 0.9) h, and the AUC(0-t) were (131.0 +/- 38.6) and (117.3 +/- 23.1) microg x mL(-1) x h, respectively. The binary IBU-ISG was successfully prepared.
Administration, Oral
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Alginates
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chemistry
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Analgesics, Non-Narcotic
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administration & dosage
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blood
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pharmacokinetics
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Animals
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Area Under Curve
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Calcium Chloride
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chemistry
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Citrates
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chemistry
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Delayed-Action Preparations
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Dogs
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Drug Compounding
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methods
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Drug Delivery Systems
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Excipients
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Female
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Glucuronic Acid
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chemistry
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Hexuronic Acids
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chemistry
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Ibuprofen
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administration & dosage
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blood
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pharmacokinetics
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Male
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Polysaccharides, Bacterial
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chemistry
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Viscosity
8.Relationship between HPVtype16/18 status and the development of cervical intraepithelial neoplasia.
Ming-tang XU ; Chun-nian HE ; Chang-tian XU ; Huan-fen ZHAO ; Shu-song WANG ; Xiu-zhi ZHANG ; Chen CHEN ; Zhi-bin HAO
Chinese Journal of Pathology 2013;42(6):400-401
Adult
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Carcinoma
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virology
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Cervical Intraepithelial Neoplasia
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virology
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Female
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Human papillomavirus 16
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isolation & purification
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Human papillomavirus 18
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isolation & purification
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Humans
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In Situ Hybridization
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Middle Aged
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Papillomavirus Infections
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Uterine Cervical Neoplasms
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virology
10.Effects of carbaryl on serum steroid hormone and the function of antioxidant system in female rats.
Yang QIU ; Jian-feng CHEN ; Ling SONG ; Jun HE ; Ru LIU ; Chun-win ZHANG ; Xin-ru WANG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2005;23(4):290-293
OBJECTIVETo investigate the effects of carbaryl on serum steroid hormone and function of antioxidant system in female Sprague Dawley rats.
METHODSCarbaryl was administrated to the adult female rats at doses of 0, 1.028, 5.140 and 25.704 mg x kg(-1) x d(-1) for 30 d. Vaginal smears of rats were taken to determine estrous cycle. Serum 17beta-estradiol (E(2)) and progesterone (P(4)) concentrations were measured by radioimmunoassay. The activities of superoxide dismutase (SOD) and glutathione-s-transferase (GST), and the levels of malondialdehyde (MDA) and glutathione (GSH) were measured by spectrophotometry.
RESULTSThe number of estrous cycle in exposed groups were obviously lower than in control group (P < 0.05, P < 0.01). Body weight gain in high dose group (25.704 mg x kg(-1) x d(-1)) was significantly lower than that in control. Meanwhile, the organ coefficient of ovary and uterus declined in a dose-dependent manner. Serum E(2) level [(19.93 +/- 2.21) nmol/L] in 25.704 mg group was lower than in control group [(28.76 +/- 6.12) nmol/L, P < 0.05], and P(4) level (1.21 +/- 0.40) nmol/L in 1.028 mg group was higher than that in control group [(0.63 +/- 0.39) nmol/L, P < 0.05]. The activity of SOD first reduced then rose in ovary, and first rose then reduced in serum. The contents of MDA increased in ovary, while decreased in the serum. GSH contents and GST activity increased in ovary, while in serum GSH contents decreased and GST activity first increased then decreased.
CONCLUSIONCarbaryl could disrupt estrous cycle and affect serum steroid hormone, and the function of antioxidant system in female SD rats.
Animals ; Antioxidants ; metabolism ; Carbaryl ; toxicity ; Estradiol ; blood ; Female ; Glutathione ; blood ; Glutathione Transferase ; blood ; Malondialdehyde ; blood ; Progesterone ; blood ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; blood