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Objective To fabricate iRGD targeted liposome-microbubble complex containing uPA ( iRGD-LMC) ,and to improve the thrombolytic efficiency and reduce the risk of thrombolysis by iRGD-LMC combining with ultrasound targeted microbubbles destruction ( UTMD ) to release drug into the thrombus site with the help of microbubble cavitation effect . Methods Biotinylated iRGD-MBs were fabricated by thin-film rehydration method .Biotinylated liposomes containing uPA were fabricated by freeze-thaw method and were conjugated to the biotinylated iRGD-MBs surface through biotin-avidin linkage . The iRGD-LMC was subjected to confocal microscopy to determine the particle morphology . The concentration , average diameter and size distribution were determined by particle sizing instrument . The uPA loading efficiency was measured by BCA Protein Assay Kit . Ultrasound imaging was performed using a Vevo 2100 ultrasound imaging system . The iRGD-LMC was irradiated by different ultrasound time and intensity to release drug . Thrombolytic effect in vitro of iRGD-LMC combined with UTMD was observed on the thrombosis model which was extracted from mouse blood . Results iRGD-LMC was successfully prepared . iRGD-LMC was exhibited a well-defined spherical morphology and homogeneous distribution ,like ordinary microbubbles . The concentration of iRGD-LMC was ( 0 .51 ± 0 .03 ) × 109 / ml and average diameter was ( 2 .62 ± 0 .12) μm . Drugs loading efficiency was ( 3878 .5 ± 97 .8) μg uPA per 108 microbubbles . iRGD-LMC could achieve contrast-enhanced ultrasound imaging in vitro . The thrombolytic effect of iRGD-LMC +US group ( 87 .66 ± 1 .69) ％ was the best in vitro ,and had significant difference with others groups ( P <0 .05) ,followed by iRGD-LMC group ( 53 .32 ± 4 .86) ％ and uPA group ( 51 .09 ± 9 .01) ％ ,Compared with PBS group ,US group ( 23 .56 ± 9 .46) ％ had thrombolytic effect . Conclusions iRGD-LMC is successfully prepared ,which has the advantages of high drug loading of liposomes and good acoustic properties of microbubbles . iRGD-LMC combined with UTMD achieves a significant thrombolytic effect in vitro .
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Objective To develop a vulnerable plaque targeting ultrasound contrast agent (UCA) and to evaluate its affinity and imaging performance in vitro.Methods E-selectin receptor-targeting UCA,which conjugated with monoclonal antibody of E-selectin,was prepared with filming-rehydration method and biotin-avidin linkage.The size and distribution of UCA were measured with particle size analyzer,the connectivity condition of microbubbles with E-selectin antibody was also detected with fluorescence analysis.The cytotoxicity from microbubble and ultrasound irradiation was evaluated through cell counting kit-8 (CCK8) assay.The adhesion effect of UCA was assessed after co-incubated with activated mouse endothelial cells (bEnd.3) and compared with that of free antibody intervention group and control group.The imaging performance of UCA at different time points was observed on an ultrasound equipment with a high-frequency transducer.Two-sample t test and one-way analysis of variance were performed to analyze the data.Results E-selectin receptor-targeting UCA was successfully prepared.The cytotoxicity result with CCK8 assay demonstrated the favorable biocompatibility of UCA.The connection amount of UCA on activated bEnd.3 cells ((6.23 ± 0.45) bubbles/cell) was significantly higher than that of the free antibody intervention group ((1.57±0.34) bubbles/cell) and control group ((0.07±0.03) bubbles/cell;F=291.43,P＜0.01).The performance of in vitro ultrasonography at the same time points showed no obvious difference between targeting UCA and control UCA (all t＜0.51,all P＞0.05).Conclusions The prepared E-selectin receptor-targeting UCA has favorable targeting and imaging capabilities.It might be a potentially ultrasound molecular imaging agent for early detection and prognosis evaluation of vulnerable plaque.
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Objective To investigate the value of ultrasonic direct signs and indirect signs in diagnosis of rotator cuff tears (RCTs).Methods Fifty-two patients underwent ultrasonography before arthroscopy were enrolled.The efficacy of ultrasonic direct and indirect signs in diagnosis of RCTs was calculated,and the accuracy of ultrasonic direct signs in diagnosis of subtypes for RCTs was compared with arthroscopic results.Results The accuracy of ultrasonic direct signs in diagnosis of the presence of tears,full-thickness tears and partial-thickness tears was 90.38％ (47/52),96.15％ (50/52)and 86.54％ (45/52),respectively.Additionally,the consistency of ultrasonic direct signs in diagnosis of subtypes for RCTs with arthroscopic results was good.With regard to ultrasonic indirect signs,the specificity of effusions including all subacromial/subdeltoid bursa effusions,intra-articular fluid and biceps tendon sheath effusions,subdeltoid bursa hernia and cartilage demarcation sign was 80.95％(17/21),90.48％ (19/21) and 95.24％ (20/21),respectively.Conclusion Ultrasonic direct signs combined with indirect signs have high clinical value in diagnosis of subtypes for RCTs.
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Objective To investigate the clinical value of high frequency ultrasound in discovering secondary pathological changes in infant with developmental dysplasia of the hip (DDH).Methods Thirtyfour infants with 36 dislocation hips were involved.Among them,32 were with unilateral dislocation of hip joint,2 were with bilateral dislocation of hip joint.High frequency ultrasound were used to observe the morphological changes of hip glenoid labrum,adipose hyperplasia in acetabulum,thickness variation of articular capsule,and development of the ossification center of the femoral head.Besides,the morphological changes of hip glenoid labrum and adipose hyperplasia in acetabul were compared with those from MRI.Results The sensitivity and specificity of ultrasound in diagnosing glenoid labral introversion were 82 ％ and 100％ respectively.The coincidence rate of diagnosing glenoid labral introversion and glenoid labral extroversion were both 83％.Adipose hyperplasias in acetabul were clearly seen in 31 hips using ultrasound.The thickness of articular capsule in the affected hip and in the healthy hip was (2.8 ± 0.6)mm versus (1.7 ± 0.4) mm,and the difference was obviously significant (P ＜0.01).The ossification center of the femoral head in the 34 infants were classified as the following four types:① which was smaller in the affected side than in the healthy side (22/34);②which was shown in the healthy side (7/34) but not in the affected side;③ which in both sides were not shown (4/34);④ which in both sides were small (1/34).Conclusions Ultrasound is a method of simple,rapid,non-invasive in diagnosing DDH,and it can be used to discover secondary pathological changes in infant with DDH,which provide important reference for clinical diagnosis and treatment.
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Objective To explore the application of high frequency ultrasonography in the rapid diagnosis of irreducible radial head subluxation in children.Methods Twenty-three patients aged 1 to 5 with unilateral radial head subluxation in children were observed by high-frequency ultrasonography and radiography,and then compared the symptomatic to the contralateral side.Results No case of radial head subluxation was found by radiography.23 cases of radial head subluxation were found by high frequency ultrasonography,10 cases with articular cavity effusion.On the diagnosis of radial head subluxation in children,the sensitivity of high frequency ultrasonography was obviously higher than that of radiography.Conclusions The sensitivity of high frequency ultrasonography in the rapid diagnosis of the radial head subluxation in children is obviously higher than that of X ray film.High frequency ultrasonography through continuous scan can determine the continuity of anatomical structure and the relationship between some views to show the location of the specific structure relations.Specifically,it can be quickly and clearly diagnosed on anterior long-axis view and the lateral coronal view of radiohumeral joint.
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<p><b>OBJECTIVE</b>To prepare a new targeted liposome ultrasonic contrast agent with anti-KDR antibody that binds specifically with KDR as the main receptor of VEGF and evaluate its physical characteristics, biological activity and specific binding capability in vitro.</p><p><b>METHODS</b>A sonicator was used to prepare the biotinylated lipid micro-bubbles (MB-B), and biotin-avidin-mediated technique was used for attachment of anti-mouse KDR monoclonal antibody to the micro-bubble shell to generate MB-BAB-KDR. MB-BAB-KDR was incubated with fluorescent second antibody to assess the link condition, and the control groups were the MB-B and micro-bubbles with the antibody alone (MB-B-KDR). A parallel plate flow chamber system was used to characterize micro-bubbles attachment efficiency to KDR Fc.</p><p><b>RESULTS</b>The surface of the micro-bubbles could carry KDR antibody through the biotin-avidin bridge and MB-BAB-KDR were spherical and well-distributed. After incubation with the second antibody, MB-BAB-KDR could be observed to emit bright green fluorescence (Grade II) as compared with little or weak fluorescence in the control MB-B group (Grade 0) and MB-B-KDR group (Grade I). Targeted micro-bubbles bound to KDR Fc increased as the KDR Fc concentration increased (P<0.05).</p><p><b>CONCLUSION</b>The targeted liposome contrast agent linked with KDR antibody by biotin-avidin bridge we prepared shows an increased binding number as the KDR Fc concentration increases, which provides a novel approach to molecular imaging study of endometrial receptivity.</p>