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

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 collagenchitosan 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.

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.

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.

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.

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* ; Cyanosis ; Diagnosis ; Early Diagnosis ; Echocardiography ; Embolism ; Female ; Femur* ; Heart Arrest ; Humans ; Critical Care ; Intensive Care Units ; Middle Aged ; Mortality ; Pulmonary Embolism* ; Vital Signs

Acute Disease ; Adult ; Altitude Sickness ; blood ; enzymology ; Glutathione ; blood ; Glutathione Peroxidase ; blood ; Humans ; Lipid Peroxidation ; Male ; Military Personnel ; Nitric Oxide ; blood ; Nitric Oxide Synthase ; blood ; Oxidoreductases ; metabolism ; Superoxide Dismutase ; blood

OBJECTIVETo investigate the expression and relationship of p27(kip1) and its related molecules Jab1 and CRM1 during proliferation of lymphoma cells U937.

METHODSU937 cells were treated with serum starvation and release, and the effects of these treatments on the cell growth was tested with cell number counting. The expression and localization of p27(kip1), Jab1 and CRM1 in U937 cells were detected by Western blot, double immunolabelling and laser scanning confocal microscopy.

RESULTSThe growth of U937 cells was blocked by serum starvation. The total protein of p27(kip1) was increased while Ser10-phosphorylated p27(kip1) -related molecules Jab1 and CRM1 were decreased. Meanwhile, the location of p27(kip1) was changed from cytoplasm into nuclei. After serum release, the location of p27(kip1) expression reappeared in the cytoplasm again.

CONCLUSIONDuring the proliferation process of lymphoma U937 cells, Jab1 and CRM1 may influence the location and expression of p27kip1, and may participate in regulation of growth of NHL cells.

COP9 Signalosome Complex ; Cell Culture Techniques ; Cell Nucleus ; metabolism ; Cell Proliferation ; Culture Media, Serum-Free ; pharmacology ; Cyclin-Dependent Kinase Inhibitor p27 ; metabolism ; Cytoplasm ; metabolism ; Humans ; Intracellular Signaling Peptides and Proteins ; metabolism ; Karyopherins ; metabolism ; Peptide Hydrolases ; metabolism ; Receptors, Cytoplasmic and Nuclear ; metabolism ; U937 Cells

OBJECTIVETo investigate the clinical value of 18F-fluorodeoxyglucose (18F-FDG) coincidence SPECT imaging versus computed tomography (CT) for malignant lymphoma in the initial staging, early response to therapy, evaluation after completion of therapy and long-term follow-up.

METHODS18F-FDG SPECT scans was performed on 61 patients with pathologically proven malignant lymphoma. The mean age of this series was 55 years ranged from 12 to 85 years. The data of these patients were retrospectively analyzed, and the result of 18F-FDG SPECT scan was compared with the CT imaging result performed within 2 weeks before or after FDG scan. 161 18F-FDG SPECT scans were performed for initial tumor staging (n=61), early response to therapy (n=42), evaluation after completion of therapy (n=26) and long-term follow-up (n=32). Each patient had a follow-up >6 months.

RESULTS(1) Compared with CT scan, 18F-FDG SPECT imaging accurately upstaged the disease for 34.4% (21/61) of these patients at initial staging. It detected the lesions which were undetected by CT including bone marrow infiltration (n=17), foci of lymph node (n=3) and liver involvement (n=1). However, 3 patients were incorrectly staged, either downstaged or upstaged by 18F-FDG SPECT imaging. Of 212 lesions in 61 patients, 18-FDG SPECT imaging detected more lesions than CT (P < 0.01). The correspondence rate of '18-FDG SPECT imaging with marrow histology was 80.3% (49/61). (2) In early evaluation of the response to treatment, the accuracy, positive predictive value and negative predictive value of 18F-FDG SPECT imaging was 85.7%, 92.0% and 76.5% respectively, which is much higher than that of CT (64.3%, 75.0% and 50.0%), therefore, the therapeutic scheme in 21.4% (9/42) of these patients was changed by 18F-FDG SPECT imaging. Of 17 cases with negative 18F-FDG SPECT scan in early evaluation of therapy, clinical remission (13 complete remission and 3 partial remission) were achieved in 16 patients. Of the 25 patients with positive ones, 13 were considered as having progressive disease. (3) In the evaluation at the end of therapy or during follow-up, 58 18F-FDG SPECT imagings were performed in 26 patients. The specificity and positive predictive value were 85.7% and 68.4% versus 59.5% and 43.3%, respectively, by CT scan. Of 14 patients with residual masses detected by CT scan, 8 were diagnosed as complete remission (CR) by 18F-FDG SPECT imaging based on persistently negative FDG scans; The other 6 were interpreted as CR (n=1), partial remission (PR, n=2), non-remission (n=1) and progressive disease (n=2), thus there was only one false-positive FDG scan in these 14 patients.

CONCLUSION18F-FDG imaging is quite effective and superior to CT scan for malignant lymphoma in initial staging, evaluation of early response to therapy and after completion of therapy, and long-term follow-up, especially for evaluating the residual masse.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Diagnosis, Differential ; Disease Progression ; Female ; Fluorodeoxyglucose F18 ; Follow-Up Studies ; Humans ; Lymphoma ; diagnostic imaging ; drug therapy ; pathology ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Neoplasm, Residual ; Radiopharmaceuticals ; Remission Induction ; Retrospective Studies ; Tomography, Emission-Computed, Single-Photon ; methods ; Tomography, X-Ray Computed ; Treatment Outcome ; Young Adult

OBJECTIVEIn order to investigate the relationship between Glutathione S-transferase M1 (GSTM1) status and the risk on colorectal cancer as well as to detect the related factors to this association.

METHODSA pooled analysis of multilevel Meta-regression was performed to estimate GSTM1 deficiency associated with the risks of colorectal cancer. Then subgroup Meta-regression was undertaken to evaluate the possible relationship between heterogeneity and the related characteristics.

RESULTSThe overall pooled odds ratios of colorectal cancer risk associated with GSTM1 deficiency was 1.17 (95% CI: 1.08-1.26). Ethnicity, percent of GSTM1 deficiency in population had significant relationships with heterogeneity across the studies (P < 0.05). Results of subgroup Meta-regression showed that GSTM1 deficiency was significantly associated with colorectal cancer risk in ethnic subgroups of Asians, Caucasians and in low level (lower than 50%) of GSTM1 deficiency population (P < 0.05). The respective pooled ORs were 1.14, 1.25 and 1.29.

CONCLUSIONGSTM1 deficiency seemed to be a risk factor for colorectal cancer, while interactions on the characteristics of ethnicity, percentage of GSTM1 deficiency in the studied population were related to this association.

Asian Continental Ancestry Group ; genetics ; Colorectal Neoplasms ; enzymology ; ethnology ; genetics ; European Continental Ancestry Group ; genetics ; Genetic Predisposition to Disease ; ethnology ; Glutathione Transferase ; deficiency ; genetics ; Humans ; Multilevel Analysis ; Odds Ratio ; Polymorphism, Genetic ; Regression Analysis ; Risk Factors