ABSTRACT
Objective To identify the effective results of ultrasound in degradation of polymeric nanoparticles released DNA .Polymeric nanoparticles was made by dehydration of polyacetylglutamicacid (PLGA, polylactic-co-glycolic acid)solution. Method Green Fluorescent Protein (GFP) was enclosed by PLGA. Different kinds of ultrasound mode and different duct cycle and power ones were used to radiate PLGA solution for 90 s, 9 min, 20 min separately after the solution prepared for 2 hrs,then putted the solution on centrifugal machine at 13000 r/m. Using Choloroform to get rid of fat-soluble impurity,then applied nanodrop to survey the releasing rate of DNA. Finally the effect of cell expression were observed by fluorescent microscope. Results The amount of DNA released from PLGA in groups which were exposed to ultrasound were significantly different from the groups which were not exposed to ultrosound. The releasing amount of former groups had upper limitation. The releasing rate was increased with the increment of the irradiation time,frequency of ultrasound;The effect of the DNA releasing and PLGA degradation by continuous-wave irradiation was stronger than pulsed-wave ultrasound. Conclusion Ultrasound can promote the degradation of PLGA, and do help in DNA releasing and expression in vitro.
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Objective To investigate the feasibility and the efficacy of ultrasound in promoting PLGA nanoparticle-mediated gene transfection in vivo.Methods Prostate cancer cell line PC-3 was used to generate xenografts in nude mice for gene transfection experiment in vivo.GFP plasmid was encapsulated in PLGA-based nanoparticles.Nanoparticles were injected into tumors locally.Two hours later,xenografts were exposed to ultrasound.Xenograft tissues were harvested in different time points to assess the efficiency of gene expression with regard to different parameters of ultrasound. Results PLGA nanoparticle-encapsulated GFP plasmids were readily transfected to PC-3 cells in vivo.A large number of GFP expressing cells were observed after exposed to ultrasound with 1.0 MHz 50% duty factor continuous wave.In comparison,ultrasound exposure with 40% duty factor pulse wave in vivo had low efficacy in terms of GFP expression.No animal death was noticed due to ultrasound exposure.Conclusions Ultrasound exposure can enhance the release of plasmid DNA content delivered by PLGA nanoparticles in vivo,local exposure to ultrasound wave would be used in conjunction with PLGA nanoparticle-mediated targeted delivery to the tissue or organ of interest.
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Objective To evaluate the effects of impaired glucose tolerance (IGT) on ventricular remodeling. Methods Parameters of every subject including left ventricular mass ( LVM), left ventricular mass index (LVMI), E/A ratio, 75 g oral glucose tolerance test (OGTT), ambulatory blood pressure monitoring(ABPM) data including 24-hour mean systolic blood pressure(mSBP) and 24-hour mean diastolic blood pressure(mDBP) were collected. Then the relationship of IGT and myocardial remodeling related parameters were analyzed. Results The rate of diastolic dysfunction was higher in the IGT combined with hypertensive group(74% ) compared with the hypertensive group( 39% )( x2 = 6. 5, P ＜ 0. 05 ). The rate of diastolic dysfunction was higher in the IGT group( 34% ) compared with the normal group( 10% ) (x2 = 5.2,P ＜0. 05). The rate of Left Ventricular Hypertrophy (LVH)in the IGT combined with hypertensive group (24%) was higher than the other three groups (Hypertension group 7%, IGT group 0, Normal group 0) (x2 =4.561,P ＜0.05), and there was no significance between the rest three groups (P ＞0.05).Stepwise multiple regression showed age and 2 Hours' Postprandial Blood Glucose were independent risk factors of E/A ratio. Conclusions These results suggested that IGT is a possible contributor to left ventricular hypertrophy and diastolic dysfunction, and is one of the histopathology of left ventricular remodeling.
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Objective To develop a clinically useful assay for detecting the mutations of HBV pre-C/BCP based on the pyrosequencing and accuracy, reproducibility and reliability of this assay was evaluated. Methods The pyrosequencing primers for HBV pre-C/BCP mutation were designed through the cluster analysis among one hundred HBV gene sequences. After the amplification of the fragment of pre-C/BCP with the template of pre-C/BCP mutation plasmids, the pyrosequencing method for pre-C/BCP detection was initially set up with this standard sample. The accuracy, reliability and reproducibility of the pre-C/BCP pyrosequencing were confirmed through the pre-C/BCP plasmids as a standard sample when compared with Sanger/genechip sequencing method pre-C/BCP pyrosequencing assay was applied for detecting pre-C/BCP mutation types of 60 chnical serum samples in HBV patients. Results The pre-C/BCP mutation detection assay based on pyrosequencing has been established in our study. The coincidence rate between pyrosequencing and Sanger squencing was 100%. The coincidence rate between the result of pyrosequencing and of genechip method was 91.7%. The reproducibility of this assay was 97. 8%. It indicates the pre-C/BCP pyrosequencing is a high-accurate method with, good-reproducibility and high-reliability. And multi-site detection can be achieved by pyrosequencing one time. A rare mutation T1758C was also detected. Conclusion Pyrosequencing for pre-C/BCP mutations assay is high-throughout method for simultaneous detection of multi-site mutation.