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ERα-36 is a novel subtype of estrogen receptor α which promotes tumor cell proliferation, invasion and drug resistance, and it serves as a therapeutic target. However, only small-molecule compounds targeting ERα-36 are under development as anticancer drugs at present. Gene therapy approach targeting ERα-36 can be explored using recombinant adenovirus armed with decoy receptor. The recombinant shuttle plasmid pDC316-Ig κ-ERα-36-Fc-GFP was constructed via genetic engineering to express an Ig κ-signaling peptide-leading secretory recombinant fusion protein ERα-36-Fc. The recombinant adenovirus Ad-ERα-36-Fc-GFP was subsequently packaged, characterized and amplified using AdMaxTM adenovirus packaging system. The expression of fusion protein and functional outcome of Ad-ERα-36-Fc-GFP transduction were further analyzed with triple-negative breast cancer MDA-MB-231 cells. Results showed that the recombinant adenovirus Ad-ERα-36-Fc-GFP was successfully generated. The virus effectively infected MDA-MB-231 cells which resulted in expression and secretion of the recombinant fusion protein ERα-36-Fc, leading to significant inhibition of EGFR/ERK signaling pathway. Preparation of the recombinant adenovirus Ad-ERα-36-Fc-GFP provides a basis for further investigation on cancer gene therapy targeting ERα-36.
Subject(s)
Adenoviridae/genetics , Cell Proliferation , Estrogen Receptor alpha/metabolism , Recombinant Proteins , TransfectionABSTRACT
Objective@#To investigate the influence of different discontinuation time of aspirin and clopidogrel before off-pump coronary artery bypass grafting (OPCABG) on postoperative bleeding and blood products transfusion requirement.@*Methods@#Three hundred and fifty-three coronary artery disease patients who underwent OPCABG from January 2017 to January 2018 at Department of Cardiac Surgery, Zhongshan Hospital, Fudan University were retrospectively analysed. There were 268 males and 85 females, aged (66.0±9.1)years. All patients were divided into three groups: (1) guideline-recommended group: patients who discontinued clopidogrel for >5 days without discontinuing aspirin before surgery; (2) without discontinuing group: patients who discontinued clopidogrel for ≤5 days without discontinuing aspirin before surgery; (3) discontinuing group: patients who discontinued clopidogrel for >5 days with discontinuing aspirin before surgery. Postoperative bleeding recorded as chest tube drainage (CTD) volume and blood products transfusion requirement and perioperative complications were recorded. CTD volumes within 12 hours after surgery between groups were compared by Mann-Whitney U tests, CTD volumes after 12 hours postoperatively were compared by repeated measures analysis of variance and blood products transfusion and complications incidence were compared by χ2 test or Fisher′s precise test.@*Results@#The 12 hours CTD volumes of guideline-recommended group, without discontinuing group, discontinuing group after surgery were 280(153) ml (M(QR)), 291(229) ml, 225(161) ml, respectively. There were no significant differences in postoperative 12 hours CTD volumes (P=0.865), red blood cells transfusion incidence (χ2=2.626, P=0.149) and fresh frozen plasma (FFP) transfusion incidence (χ2=1.258, P=0.324) between guideline-recommended group and without discontinuing group. However, the 12 hours CTD volumes were significantly higher in guideline-recommended group patients compared with disconutinuing group patients (U=5 247, P=0.002). No significant differences were observed in red blood cells (χ2=0.182, P=0.757) and FFP (χ2=0.083, P=0.839) transfusion rate between these two groups. Repeated measures analysis of variance indicated that when patients began to take antiplatelet drugs (aspirin and clopidogrel) after 12 hours postoperatively, the change of CTD volumes beyond 12 hours after surgery didn′t differ either between guideline-recommended group and without discontinuing group (F=0.019, P=0.941) or between guideline-recommended group and discontinuing group (F=2.447,P=0.113). Besides, the incidence of perioperative arrhythmia was significantly higher in guideline-recommended group patients compared with without discontinuing group patients (4.8% vs. 0, χ2=5.073, P=0.038).@*Conclusions@#OPCABG patients who discontinued aspirin before surgery had lower postoperative 12 hours CTD volumes but similar blood products transfusion rate and CTD volumes beyond 12 hours postoperatively compared with patients adhering to the current guideline-recommended protocol. And for patients who discontinued clopidogrel for ≤5 days, postoperative CTD volumes and blood products transfusion requirement were similar but the incidence of perioperative arrhythmia was significantly lower compared with guideline-treated patients.
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Objective To detect the expression of ITGB1 in the serum and placenta of pregnant women with preeclampsia and its clinical significance. Methods A total of 150 pregnant women with sever or mild pre-eclampsia and normal pregnant women were collected. The expression change of ITGB in the serum and placenta was detected by real-time PCR. The correlation between the expression of serum ITGB1 and clinical symptoms in normal pregnant women and pregnant women with preeclampsia was explored. Results There was significantly high expression of ITGB1 in the serum and tissue of patients with preeclampsia and placenta and the expression of ITGB1 in patients with preeclampsia and normal pregnant women was positively correlated (P = 0.011). The expression level of serum ITGB1 was positively correlated with systolic blood pressure,diastolic blood pressure,24 h urine protein and vascular disease. Serum ITGB1 was used as a diagnostic marker for preeclampsia,with an AUC of 0.751 ,a specificity of 81% and a sensitivity of 75%. Conclusion ITGB1 is involved in the occurrence and development of preeclampsia,and is positively correlated with clinical symptoms. The expression of serum ITGB1 can be used to predict the occurrence of preeclampsia.
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Objective To detect the expression of ITGB1 in the serum and placenta of pregnant women with preeclampsia and its clinical significance. Methods A total of 150 pregnant women with sever or mild pre-eclampsia and normal pregnant women were collected. The expression change of ITGB in the serum and placenta was detected by real-time PCR. The correlation between the expression of serum ITGB1 and clinical symptoms in normal pregnant women and pregnant women with preeclampsia was explored. Results There was significantly high expression of ITGB1 in the serum and tissue of patients with preeclampsia and placenta and the expression of ITGB1 in patients with preeclampsia and normal pregnant women was positively correlated (P = 0.011). The expression level of serum ITGB1 was positively correlated with systolic blood pressure,diastolic blood pressure,24 h urine protein and vascular disease. Serum ITGB1 was used as a diagnostic marker for preeclampsia,with an AUC of 0.751 ,a specificity of 81% and a sensitivity of 75%. Conclusion ITGB1 is involved in the occurrence and development of preeclampsia,and is positively correlated with clinical symptoms. The expression of serum ITGB1 can be used to predict the occurrence of preeclampsia.
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Objective: To study the acute toxicity of Xiaobai capsules in mice after intragastric administration .Methods: The mice were randomly divided into two groups , the treatment group and the control group .The treatment group was given Xiaobai cap-sules by gavage, 3 times daily.The acute toxicity was recorded, and the median lethal dose (LD50) and the maximum dose were deter-mined.Results:The maximum daily dose of Xiaobai capsules was 141.6 g· kg-1(equivalent to 211.3 times of the clinical dose).At the dose, the mice showed no toxicity without death in 14 days or changes in organs after the dissection .Conclusion:Xiaobai capsules have very low acute toxicity in mice after intragastric administration with high security .
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Objective To establish a reversed-phase high performance liquid chromatograph ( RP-HPLC ) method for determination of equilibrium solubility and oil/water partition coeficient of phloridzin in different solvents. Methods A RP-HPLC method was established to detect the concentration of phloridzin in water and different organic solvents. The partition coefficients in the n-octanol-water/buffer solution systems of phloridzin were determined by shaking flask method. The Inertsil ODS-3 (4.6 mm×150 mm, 5μm) column was used and the detection wavelength was 284 nm.The flow rate was 1.0 mL??min-1, and acetonitrile-0.05% phosphoric acid(30??70)was used as mobile phase. Results The equilibrium solubility of phloridzin was 2.07 mg??mL-1 in water and 838.63 mg??mL-1 in methanol at 25 ℃.A good linear relationship of phloridzin was obtained within the range of 0.054 9-1.098 0 μg.The regression equation was Y=2 152.9X+7.26 (r=0.999 9).The solubility values of phloridzin were higher in ethanol and propylene glycol than in other solvents. Conclusion RP-HPLC method is simple, quick and accurate for the determination of phloridzin.Phloridzin was almost insoluble in petroleum ether and poorly soluble in water.The equilibrium solubility is higher in methanol than in other solvents. The apparent distribution coefficient of phloridzin varies significantly with pH under the alkaline conditions but less in the acidic solution.
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Objective To analyze tissue vascular endothelial growth factor receptor 1 (VEGFR-1)and VEGFR-2 and their soluble form (sVEFGR-1 and sVEGFR-2) in the plasma in patients with preeclampsia,and to explore its clinical significance.Methods sVEGFR-1 and sVEGFR-2 expressions in plasma of normal pregnancy women and preeclampsia patients were detected by enzyme-linked immunosorbnent assay (ELISA) ; VEGFR-1,VEGFR-2,sVEGFR-1 and sVEGFR-2 mRNA expressions of normal pregnancy women and preeclampsia patients in placenta were detected by reverse transcription-polymerase chain reaction (RT-PCR).Results ELISA results showed that the level of sVEGFR-1 in plasma of normal control group before and after delivery was (12.33 ± 1.52) ng/ml and (4.55 ± 0.31) ng/ml,respectively,while that of preeclampsia group was (77.25 ± 9.47) ng/ml and (8.13 ± 0.74) ng/ml,respectively; the differences between before and after delivery in the two groups were of statistical significance.The level of sVEGFR 2 in plasma of normal control group before and after delivery was (8.74 ± 1.24) ng/ml and (6.43± 0.55) ng/ml,respectively,while that of preeclampsia group was (5.69 ± 0.75) ng/ml and (4.96 ±0.67) ng/ml,respectively.RT-PCR results were consistent with ELISA results.Conclusions Rapid decrease of plasma sVEGFR-1 and continuously low-level expression of plasma sVEGFR-2 indicated that VEGFR-1 might be closely related to preeclampsia,and decrease of plasma sVEGFR-2 in preeclampsia women might be taken as a marker of endothelial cell function disorder.
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OBJECTIVE:To determine the concentration of borneol in blood after keeping Suxiao Jiuxin pills in healthy men's mouth by GC METHODS:HP-5 quartz elasticity capillary column(15m?0 53mmID) was used with nitrogen as the carrier gas,temperature of column was 85℃ The injector temperature and detector temperature were 180℃,200℃ respectively Naphthalene was used as internal standard RESULTS:The standard curve was linear within the concentration range of 50～1 000ng/ml of borneol,the calibrating equation was Y=-0 1 273+0 0 015X The recovery was (98 7?2 31)%,(RSD=2 76%,n=6) The peak of blood drug concentration was obtained in 15 min or so CONCLUSION:The method was simple,rapid,accurate and suitable for pharmacokinetic study of preparation containing borneol
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0.05). Conclusion After heated, the physical stability of UAE and UAS is reduced, the viscosity become lower, ADM releasing rate is fell. The heated Lipiodol-Adriamycin pharmaceutics had advantage in the interventional embolization chemotherapy of the neoplasm.