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Objective:To prepare a dual targeting nanobubble both for HER2 target and cancer cell target, and compare the targeting ability with single targeting nanobubble either with HER2 targeting or cancer cell targeting ability in vitro.Methods:Nanobubbles(NBs) were prepared using the modified thin film hydration method and then connected with IR783 directly (NBs-IR783), HER2 antibody Affibody by avidin-biotin method (NBs-Affibody), or both of them (IR783-NBs-Affibody). The size distribution, stability, and biosafety of these contrast agents were observed. Flow cytometry and immunofluorescence were applied to compare the binding rate of these NBs against HER2 positive breast cancer cell in vitro.Results:The average size of NBs-Affibody, NBs-IR783 and IR783-NBs-Affibody was (538.4±95.8)nm, (551.8±114.8)nm, and (482.7±54.3)nm, respectively. The binding rate for HER2 positive cell was 26.6%, 97.6%, 84.5%, while for HER2 negative cell was 5.4%, 99.9% and 99.3%, respectively. The results of dual-labeled flow cytometry showed that the binding rate of IR783-NBs-Affibody mediated by IR783 to HER2 positive breast cancer cells were 79.5% and Affibody mediated HER2 targeting binding rate were 19.4%. However, NBs-IR783 only showed IR783-mediated tumor targeted binding without HER2 targeting ability with binding rate of 2.3%.Conclusions:The prepared IR783-NBs-Affibody has the dual targeting ability for HER2 positive breast cancer cell, which is superior to the single targeting NBs (NBs-Affibody and NBs-IR783). IR783-NBs-Affibody is optimal to meet the imaging requirements of tumor heterogeneity.
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Objective To research the Annexin V-nanoscale ultrasound contrast agents'preparation, ultrasound imaging and the ability to binding apoptosis cells of tumor in vitro.Methods The nanoscale bubble (Nanobubbles,NBs ) packaged the octaflouropropane (C3 F8 ) gas was prepared by thin film hydration.The Annexin V-Nanobubbles (AVNBs ) solutions was acquired through conjugating the biotinylated-Annexin V to the surface of the NBs by biotin-streptavidin bridging chemistry.The size and zeta potential of AVNBs were measured by NanoPlus-3 zeta/nano particle analyzer.The shift in size distribution of AVNBs bubbles was analyzed for the stability,after it was stored at 4 ℃ for different time. AVNB's shape were measured by scanning electron microscopy.The AVNBs bubble was measured using an ultrasound system for echogenicity in vitro,and SonoVue was for control.Finally,the ability of AVNBs binding with apoptosis cells of tumor in vitro was determine via the fluorescence microscope.Results AVNBs has a size distribution of (640.2±32.1 )nm,and a mean zeta potential of (-23.30 ±5.71 )mV.Its size remained relatively constant and appeared to show less size variation within the 24 h analysis period. AVNBs solutions were visible milky white and slightly suspension liquid with the naked eye.Under scanning electron microscopy,AVNBs were uniform hollow sperical cavitation bubble with small size and larger dispersibility in solution.The AVNBs and SonoVue solution had the same higher grayscale signal intensity by ultrasonic imaging.The AVNBs binded well with apoptosis cells of tumor in vitro,and the rate of binding was (97.55 ± 1 .30 )%.Conclusions The AVNBs particles prepared by method of thin film hydration have a nanoscale size,good stability and echogenicity.It can be targeted binding with the apoptosis cells of tumor in vitro.
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Objective To explore the value of superior vena cava (SVC) Doppler spectrum in evaluation of the therapeutic effects of intermittent hypobaric hypoxia (HBHO)-induced pulmonary hypertension (PH) in rats and establish a new method for assessing pulmonary hypertension.Methods Fifty male SD rats were included.Forty of them were developed HBHO-PH and randomly divided into four groups with different manipulations:beraprost treatment group,sildenafil treatment group,placebo group and model group,with 10 rats in each group.The rest of 10 rats served as controls.Cardiac structure,pulmonary peak velocity (PAVmax),tricuspid peak velocity (TVEmax) and SVC Doppler ultrasound were performed in all rats before and after treatment for 2 weeks.Pulmonary arterial pressure was determined by the right heart catheterization.The relationship of the parameters with the pulmonary arterial pressure was analyzed.HE staining was done in lung tissues.Results Right heart catheterization showed that pulmonary artery systolic pressure (PASP) of the treatment groups were lower than that of the model group,the difference was significant(P ＜0.01).Pulmonary artery pressure decreased in varying degrees both after placed in a normal environment and after 2 weeks of treatment.PASP of the beraprost treatment group and the sildenafil treatment group decreased significantly and showed no statistical significant difference with the control group.In contrast,PASP of the placebo group was significant higher than that of control group(P ＜ 0.05).No statistically significant difference was found between the non-placebo treatment groups in PASP.Compared with the control group,the right ventricle/body weight ratio of the model group was statistically significant increased in all other groups(P ＜0.01).Right SVC AR/S ratio was well correlated with PASP (r =0.603,P =0.001).Right SVC AR/S ratio was statistically significant lower in treatment groups compared with the model group (P ＜0.01).All groups except the control group demonstrated various degrees of pulmonary arterial wall thickening.Conclusions The right SVC spectrum of AR/S correlates well with PASP and can be used to evaluate the pharmacological therapeutic effect of rats with HBHO-PH,in particular with moderate to severe PH.