Experiment of nanobubbles-mediated plasmid green fluorescent protein(GFP) transfer into skeletal muscle in mice / 中华超声影像学杂志

ObjectiveTo explore the effect of nanoscale bubbles transfering gene in skeletal muscle cells in mice.MethodsPlasmid DNA encoding green fluorescent protein (GFP) was mixed with bubbles dissolved in saline and injected into the tibialis anterior (TA) muscle of C57B10 mice with and without ultrasound (US).The ultrasound frequency was 1 MHz and the pulse repetition frequency was 100 Hz with 20％ duty cycle.The spatial peak temporal peak intensity (ISPTP) power level was 2 W/cm2.The entire treatment period was 30 seconds.The efficiencies of GFP transgene expression were determined under different experinmental conditions.Mice were sacrificed 1 week after plasmid DNA injection.Fibres with fluorescence green signals were determined as GFP-positive fibres by fluorescence microscopy.Readout was performed on the section with the maximum number of transfected fibers.Results1 )Albumin nanobubbles:in the conditon of with or without ultrasound,albumin nanobubbles had significantly increased gene expression compared with negative control (P ＜0.05),but significantly decreased gene expression compared with positive control ( P ＜ 0.05 ).2) Phospholipid nanobubbles:In the conditon of without ultrasound,there were no significant differences compared with negative and positive control ( P ＞0.05).In the conditon of with ultrasound,phospholipid nanobubbles had significantly increased gene expression compared with negative control ( P ＜0.05).No significant difference was observed between phospholipid nanobubbles and positive control ( P ＞ 0.05).ConclusionsNanobubbles could enhance gene transfection efficiency for skeletal muscle fibres in mice.Nanobubbles with perfluoropropane gas and albumin have potentiality in gene-enhanced transfection.

The experiment in the mechanisms of microbubble-mediated GFP gene enhancement in skeletal muscle in C57B10/mdx mice / 中华超声影像学杂志

Objective To investigate the role of sonoporation and the deblic of microbubbles with perfluoropropane gas and albumin in the mechanisms of microbubble-mediated gene enhancement by experimenting in skeletal muscle in C57B10/mdx mice. Methods Plasmid DNA (10 μg) encoding green fluorescent protein (GFP) was mixed with Optison or SonoVue dissolved in saline and injected into the tibialis anterior (TA) muscle of /C57B10/mdx mice with and without adjunct ultrasound. The efficiencies of GFP transgene expression were determined under different experimental conditions. C57B10 mice as normal control:①C57B10 mice + saline (4 left TAs);②C57B10 mice + saline + ultrasound (4 right TAs) ;③C57B10 mice + Optison(4 left TAs);④C57B10 mice+ Optison + ultrasound(4 right TAs);⑤ C57B10 mice + SonoVue(4 left TAs) ;⑥C57B10 mice + SonoVue + ultrasound(4 right TAs). Mdx mice groups:① mdx mice + saline(4 left TAs) ;② mdx mice + saline + ultrasound(4 right TAs);③ mdx mice + Optison (4 left TAs) ; ④ mdx mice + Optison + ultrasound (4 right TAs); ⑤mdx mice + SonoVue(4 left TAs) ;⑥mdx mice + SonoVue + ultrasound(4 right TAs). Mice were sacrificed 1 week after plasmid DNA injection. Fibres with fluorescence green signals were determined as GFP-positive fibres by fluorescence microscopy. Readout was performed on the section with the maximum number of transfected fibers. Results C57B10 mice: ?Optison without ultrasound had significantly increased gene expression compared with negative control ( P <0. 01). SonoVue without ultrasound did not enhance gene expression. ?Optison with ultrasound had significantly increased gene expression compared with negative control (P < 0.01). ?SonoVue with ultrasound had significantly increased gene expression compared with negative control ( P<0. 01).Mdx mice:? Compared with C57B10 mice, GFP alone demonstrated significant GFP expression in mdx mice ( P <0. 01) , Optison demonstrated significant GFP expression in mdx mice ( P <0.01), and SonoVue demonstrated significant GFP expression in mdx mice ( P <0. 01). ?Microbubble groups (Optison and SonoVue) had significantly increased gene expression compared with negative control (P <0. 01). Conclusions In the mechanisms of microbubble-mediated gene enhancement, sonoporation is the key step. The deblic of microbubbles with perfluoropropane gas and albumin is the main constituent in the mechanisms of Optison-mediated gene enhancement. fibers.Results C5781 0 mice:①Optison without ultrasound had significantly increased gene expressioncompared with negative control(P<0.01).SonoVue without ultrasound did not enhance gene expression.②Optison with ultrasound had significantly increased gene expression compared with negative control(P<0.01).③SonoVue with ultrasound had significantly increased gene expression compared with negativecontr01(P

Quantitative echo-tracking technique for evaluating carotid arterial elasticity function in patients with systemic lupus erythematosus / 中华风湿病学杂志

Objective To explore the clinical value of early diagnosis of atherosclerosis in patients with systemic lupus erythematosus (SLE) using vascular echo-tracking technique and to detect changes of elastieity of carotid artery quantitatively in SLE patients.Methods Fifry patients with SLE were divided into SLE1 group(duration≤1 year),and SLE2 group(duration>1 year)based on different course.An ultrasonic echo-tracking method was used to measure patients'pressure strain elastic modulus (Ep),the stiffness constant(β),arterial compliance(AC),augmentation index(AI),pulse wave velocity (PWVβ) and intimamedia thickness(IMT)of the common carotid arteries in 50 patients with SLE and in 25 healthy control subjects.Results Among carotid artery elasticity indicators of three groups,there was no significant difference in AI(P>0.05).Ep,β,PWVβ parameters of SLE1 group and SLE2 group were statistically higher than that of the control group[Ep of SLE1 group,SLE2 group,control group was (69±20),(103±40),(48±18)kPa respectively;β was 5.2±1.9,8.0±3.1,4.2±1.3 respectively;PWVβ was 5.2±0.7,6.3±1.1,4.5±0.7]respectively,but AC(AC of SLE1 group,SLE2 group,control group was(1.1±0.3),(0.8±0.3),(1.2±0.6)mm2/k respectively]lower than the controls(P<0.01).Ep,β,PWVβ in SLE2 group was significantly increased compared with SLE1 group,but AC was decreased (P<0.01).Conclusion The application of echo-tracking technology can be used to diagnose early atherosclerosis.Complications of cardiovascular disease in SLE have high clinical value.

Effects of stellate ganglion block in the pulmonary artery endothelial nitric oxide synthase and pulmonary artery pressure in rabbits with hypoxic pulmonary hypertension / 中华麻醉学杂志

Objective To investigate the effects of stellate ganglion block(SGB) on the pulmonary artery endothelial nitric oxide synthase (eNOS) and mean pulmonary artery pressure ( MPAP) in rabbits with hypoxic pulmonary hypertension ( HPH) .Methods Twenty-four rabbits of both sexes weighing 2.5-3.0 kg were anesthetized with 1.5% sodium pentobarbital 30 mg? kg-1 . Spontaneous breathing was maintained. Left stellate ganglion was exposed aseptically. An epidural catheter was fixed with one end placed close to stellate ganglion and the other end outside the neck through a hole in the skin for administration of drugs. One week later the rabbits were randomly divided into 4 groups with 6 animals in each group: Ⅰ control group; Ⅱ SGB group; Ⅲ hypoxia group and Ⅳ SGB + hypoxia. In group Ⅲ and Ⅳ the animals were placed in a closed box filled with hypoxic air (O2 % = 10 ? 2% ) 8 h a day for 2 weeks. In group Ⅱ and Ⅳ 0.25% bupivacaine 0.5 ml was injected through the catheter 3 times a day for 3 days. In group Ⅰ and Ⅲ normal saline 0.5 ml was injected instead of bupivacaine. The effect of SGB was confirmed by ptosis and miosis. The content of eNOS in pulmonary artery was detected using immunohistocheistry technique. Pulmonary artery was cannulated after thoracotomy for determination of MPAP. Results There was no significant difference in MPAP between control and SGB groups. MPAP was significantly increased in hypoxia and hypoxia + SGB groups compared with control group(P