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Biliary atresia (BA) is the most common cause of persistent jaundice in infant,it can rapidly advance to biliary cirrhosis and death.Surgical treatment in time can decrease mortality,so it is essential to be differentiated from other diseases as early as possible.Main diagnosis methods of BA include clinical feature of patients,liver biopsy,and high-frequen cy ultrasonography,MR cholangiopancreatography (MRCP),hepatobiliary scintigraphy.High-frequency ultrasonography is superior to other methods with high diagnostic specificity and sensitivity.The progresses of ultrasonography in diagnosis of BA were reviewed in this article.
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Objective To evaluate the differential diagnostic value and superior of biliary atresia(BA)in the infants with cholestatic hepatopathy by high-frequency ultrasonography (HUS).Methods After 4 hours fasting,124 infants with cholestatic hepatopathy were scanned with high-frequency US.The data of hepatic size and parenchyma,gallbladder,triangular cord (TC) sign,bile duct,right hepatic artery (RHA)and portal vein (PV) were observed and measured.Meanwhile,the other data were collected,which included the clinical diagnosis,blood biochemical tests,the MRCP and dynamic duodenal liquid color check finding,the pathological results after liver puncture biopsy and so on.Results In 124 infants with cholestatic hepatopathy,BA was found in 61 infants and ruled out in 63.TC thickness,RHA diameter,and gallbladder length and width exhibited significant differences between the group with BA and the group non-BA(all P ＜0.001).The correctness for the diagnosis of BA was 90.3％ by the combination of TC sign and abnormal gallbladder morphology,and 83.1 ％ by stool color,81.5 ％ by γ-GT,47.5 ％ by MRCP,83.3 ％ by dynamic duodenal liquid color check,95.2％ by the pathology after liver puncture biopsy,respectively.Conclusions HUS is superior to other diagnostic methods in BA with higher accuracy rate,noninvasion,simplicity and economy.
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Objective To investigate the optimal ultrasound exposure parameter on H22 neoplasms of mice meditated by ultrasound exposure combined with self‐made nanobubbles ,and then observe their therapeutic effect combined with cisplatin and their possible mechanism of anti‐tumor . Methods Thirty mice engrafts models with subcutaneous H22 neoplasms were established and divided into 6 groups randomly ,which received ultrasound exposure at different intensity and exposure time . The contrast enhanced ultrasound imaging ( CEUS ) was performed in every group at the four time points of before treatment and at 0 h ,24 h ,72 h after treatment . To obtain the optimal ultrasound parameters ,the tumor inhibitory effect was assessed by enhanced intensity ( EI) and microvascular density ( MVD) . The H22 tumor were treated by ultrasound exposure nanobubbles combined with cisplatin to observe their tumor growth inhibition rate ,and the microvessels density and nuclear associated antigen Ki‐67 proliferation index were measured by immunohistochemical staining . Results There was a statistically difference in enhanced intensity (EI) between the experimental groups and control group ( P < 0 .05) . With the increasing of ultrasound intensity and exposure time ,the tumor inhibitory effect was more obvious ,with an increasing side reactions . Except the simple ultrasound group ,there was a statistical difference in tumor inhibition ,the mean MVD and the tumor cell proliferation index (KI‐67) between control group and the other ultrasound therapy groups ( P<0 .05) . The tumor inhibitory rate was the highest ( tumor inhibition rate 70 .0% ) and the mean MVD and KI‐67 expression were the lowest ( P <0 .05) in the combination group comparing with the others . Conclusions The ideal ultrasound exposure parameter of tumor inhibition showed that exposure intensity chose 1 W/cm2 and exposure time chose 1 min or 3 min intermittence . The ultrasound exposure self‐made nanobubbles combined with cisplatin could enhance the tumor inhibitory effect .Its mechanism may be related to the decrease of microvascular density ,the inbition of tumor cell proliferation and the increase of tumor cell necrosis .
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Objective To summarize the sonographic characteristics of high frequency ultrasound and elastography of thyroid microcarcinoma(TMC),and to analyze the causes of misdiagnosis.Methods The preoperative ultrasonic data of 245 suspicious TMCs in 202 patients,as confirmed by operation pathology,were retrospectively analyzed.Results Preoperative ultrasonography accurately diagnosed 221 TMCs,the diagnosis rate was 90.2％,and the misdiagnosis rate was 9.8％.Among the missed diagnosed lesions,18 lesions were nodular goiter,others were 3 nodular goiter with focal papillary hyperplasia of follicular epithelium,2 nodular goiter with adenomatous hyperplasia,1 focal lymphocytic thyroiditis,respectively.The thyroid lesions≤0.5 cm in diameter were more easily to misdiagnose.According to the importance of the ultrasonographic features of TMC,the order were aspect ratio (A/T) ≥ 1,irregular-shape,microcalcifications,low or very-low echo.Based on the above corresponding characteristics and considering other features together,the diagnostic accuracy rate were 94.1 ％,93.9％,92.4％ and 90.5％,respectively.Less blood supply and ill-defined boundary were the secondary sonographic signs of TMC.The elastographic scores of TMC were most showed 4 to 5 points.Diagnosis of TMC relied on elastography alone is less effective,but when elastograph diagnosis based on high frequency ultrasound,the diagnostic accuracy is much higher,especially when there is no calcification in the lesions.Conclusions High-frequency ultrasound has a very important value in the diagnosis of TMC,while elastography has certain assistant value on the basis of high-frequency ultrasonic diagnosis.
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Objective To investigate whether ultrasound-mediated microbubble destruction(UMD) could enhance cationic liposome (CL) induced plasmid DNA delivery or not,and optimize the transfection conditions.Methods Multiple parameters were explored to obtain the optimal transgene efficiency by means of with or without serum in culture medium,various CL or nano-liposomal bubble(NB) concentrations,different time point of ultrasonic irradiation.The transfection efficiency was assessed by fluorescence microscopy and flow cytometer,and cell viability was evaluated by Cell Counting Kit-8 (CCK-8) assay.Results The serum could protect the cells but show little impact on transfection efficiency induced by CL.CL and plasmid DNA at a weight ratio of 4 ∶ 1 exhibited high transfection efficiency of (17.71-± 0.79)％ and high cell viability of (91.28 ± 0.76) ％.CL combining with ultrasonic irradiation at the time point of 1 hour could increase the transfection efficiency to (24.85 ± 0.78)％ (P ＜0.01).Higher transfection rate (32.47 ± 4.01) ％ was obtained by adding NB at the concentration of 10 ％ (P ＜0.05).Conclusions UMD accompanied with CL could enhance gene delivery effectively,which would provide a new method for gene therapy.
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Objective To observe self-made cationic nanobubbles as non-viral gene carrier to transfer green fluorescent protein reporter gene into HepG2 cell in vitro.Methods Cationic nanobubbles(PNB) were prepared by sonicating liposomes、polyethylenimine and perfluoropropan.The surface potential and the size of nanobubbles were assessed by laser particle analyzer.HepG2 cells were incubated with DNA,nanobubbles with or without ultrasound exposure.The transfection efficiency was evaluated by flow cytometer and the cell viability by cell counting Kit-8.Results The mean diameter of PNB was (834.57 ± 6.4) nm and the surface charge was (4.15± 0.98)mV.The PNB-DNA complexes,which blocked by the Agarose gel electrophoresis,could effectively transfer HepG2 cells,and the ultrasound exposure could enhance the transfection efficiency further significantly (P ＜ 0.05).Conclusions The new PNB could effectively combine with pDNA to enhance gene delivery and ultrasound exposure could improve its efficiency further in HepG2 cell in vitro.
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Objective To made a kind of nano-liposomal bubbles(NB) and observe its base physical properties and ultrasound contrast enhanced effect,and compared with SonoVue in vitro and in vivo.Methods The liposome was made with the method of reverse phase evaporation,and then the liposome was sonieated to prepare NB.After the physical properties of NB such as its morphous,average particle size,surface potential and concentration were observed and calculated,200 μl of saline and two contrast agents (NB and SonoVue) were injected into degassed PBS (10 ml) respectively to observe their enhancing effects.For in vivo study,a healthy rabbit heart and liver were imaged before and after intravenous injection of 2.0 ml/kg of saline and two contrast agents in succession to compare their dynamic enhancing effects in the grey scale imaging subjectively.Results The self-made NB distributed uniformly and its size ranged from 133.1 ～199.5 nm with a mean diameter of (171.60 ± 30.82) nm.Its surface potential and concentration were -(1.92± 0.65)mV and (3.8 ～ 5.6) × 108/ml individually.These basic characteristics were not observed changing dramatically after placement one week to one month under room temperature.The NB and SonoVue all displayed significantly enhancing effect comparing with saline no matter in vitro or in vivo.Conclusions The self-made NB are stable and distribute uniformly,which display the same contrast enhancing effect as SonoVue in vitro and in vivo.The self made NB would have a more potential use in ultrasound molecular imaging and gone or drug delivery.
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This study examined the effect of P85 (a pluronic block copolymer) and microbubble (MB) ultrasound contrast agents under ultrasound irradiation on gene transfection and expression. The pEGFP plasmids that can encode enhanced green fluorescent protein (pEGFP) served as a report gene and were mixed with different concentrations of MB/0.05% (w/v) P85. Then the plasmids were transfected into human hepatoma G2 (HepG2) cells. The HepG2 cells treated with MB/P85 or without treatment were exposed to ultrasound (US parameters: 1 MHz, 1.0 W/cm(2), 20 s, 20% duty cycle). Twenty-four hours later, the transfection efficiency was assessed by fluorescence microscopy and fluorescence activated cell sorting (FACS) analysis. The cell viability was evaluated by Trypan blue exclusion test. The results showed that the gene transfection efficiency in HepG2 cells under ultrasound irradiation was significantly higher than that without ultrasound irradiation. HepG2 cells in the MB or P85 group in the absence of ultrasound expressed less amount of green fluorescent protein. The expression efficiency reached (22.14±3.06)% and the survival rate was as high as (55.73±3.32)% in the 30% MB plus P85 group. It was concluded that MB and P85 in the presence of ultrasound can enhance gene transfection and expression.
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Objective To explore the effects of P85,microbubbles and ultrasound on plasmid DNA skeletal muscle gene transduction of mice in vivo. Methods Plasmid encoding green fluorescent protein (GFP) ,which conjugated with 0.05% P85 and/or microbubbles, 10% Optison,was injected into the tibialis anterior(TA) muscle of mice with or without ultrasound irradiation (1 MHz, 1 W/cm2 2 min,20% duty cycle). Mice were killed 1 week after injection. The TA muscles were removed and snap-frozen immediately in isopentane cooled by liquid nitrogen and sections 7 μm thick were cut at intervals. One set of sections mounted with DAPI were used to assess the transfection efficiency by counting the number of GFP-positive fibers under fluorescence microscopy,and the other set of sections were stained with haematoxylin and eosin to assess the tissue damage area. Results The P85 and Optison significantly enhanced the plasmid DNA skeletal muscle gene delivery in vivo separately (P<0.01, P<0.05).Ultrasound exposure could significantly enhance the efficiency of P85 induced gene delivery(P<0.01) but not of Option(P>0.05).The gene delivery efficiency induced by P85 was higher than that by Optison no matter with or without ultrasound irradiation(P<0.01). When the P85 conjugated with Optison, they could further significantly enhance gene delivery efficiency with ultrasound exposure (P<0.01). Meanwhile, ultrasound exposure could increase the muscle damage areas in the groups with microbubbles (P<0.01). Conclusions The P85,microbubbles and ultrasound exposure display synergistic effect to enhance plasmid DNA transduction in skeletal muscle of mice in vivo.
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The value of color Doppler flow imaging (CDFI) and intravenous contrast-enhanced ultrasound (CEUS) for assessing the transplanted liver and early diagnosing complications by examining hemodynamic changes was discussed. Seventy-five patients with orthotopic liver transplantation (OLT) underwent CDFI. The following parameters were measured: peak systolic velocity (PS), resistance index (RI) and Doppler perfusion index (DPI) of the hepatic artery (HA), time average velocity (TAV) of portal vein (PV) and velocity of hepatic vein (HV) in different stages postoperation. And 11 patients of them received CEUS. Thirty healthy subjects were enrolled as controls. The results showed that: (1) In 23 patients without obvious complications, TAV of PV within 15 days post-operation was significantly higher than in controls (P<0.05), PS and DPI of HA within 7 days postoperation were lower, but RI was higher than in controls (P<0.05); (2) When the hepatic artery thrombosis (HAT) occurred, PS and DPI of HA were obviously decreased, but TAV of PV significantly increased like a high saw-tooth wave; (3) While rejection occurred, both TAV of PV and PS of HA were decreased with the increase in RI of HA, and the triphasic wave of HV disappeared and displayed as saw-tooth wave; (4) The incidence of biliary complications in liver transplantation was increased when DPI was reduced; (5) Seven cases of hepatic carcinoma relapse after OLT demonstrated hyperecho in the arterial phase and hypoecho in the portal and later phase on CEUS; (6) In 2 cases of HA thrombus, there was no visualized enhancement in arterial phase of CEUS, but enhancement during the portal vein and parenchymal phase. It was concluded that the hemodynamic changes of PV, HA and HV in the transplanted liver are valuable for assessing the transplanted liver and early diagnosing complications on CDFI and CEUS.
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Bile Ducts/pathology , Contrast Media/administration & dosage , Hemodynamics , Hepatic Artery/pathology , Infusions, Intravenous/methods , Liver Transplantation/adverse effects , Liver Transplantation/methods , Liver Transplantation/diagnostic imaging , Perfusion , Postoperative Complications , Ultrasonography/methods , Ultrasonography, Doppler/methodsABSTRACT
To examine the role of ultrasound in gene delivery in vitro, three cells lines were exposed to the low-frequency ultrasound of varying intensities and for different durations to evaluate their effect on gene transfection and cell viability of the cells. Microbubble (MB), Optison (10%), was also used to observe the role of the microbubbles in gene transfection. The results demonstrated that as the ultrasound intensity and the exposure time increased, the gene transfer rate increased and the cell viability decreased, but at high energy intensities, the cell viability decreased dramatically, which caused the transfer rate to decrease. The most efficient ultrasound intensity for inducing gene transfer was 1 W/cm2 with duration being 20 s. At the same energy intensity, higher ultrasound intensity could achieve maximal gene transfer rate earlier. Microbubbles could increase ultrasound-induced cell gene transfer rate by about 2 to 3 times mainly at lower energy intensities. Moreover, microbubbles could raise the maximum gene transfer rate mediated by ultrasound. It is concluded that the low-frequency ultrasound can induce cell gene transfer and the cell gene transfer rate and viability are correlated with not only the ultrasound energy intensity but also the ultrasound intensity, the higher ultrasound intensity achieves its maximal transfer rate more quickly and the ultrasound intensity that can induce optimal gene transfer is 1 W/cm2 with duration being 20 s, and microbubbles can significantly increase the maximal gene transfer rate in vitro.
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In order to assess whether gene transfection could be mediated by ultrasound in associa- tion with P85 and find the appropriate parameters of ultrasound irradiation, the effects of ultrasound with or without P85 on gene transfection of HepG2 cells were examined. The HepG2 cells were irra- diated by ultrasound at 1 MHz, 0.4-2.0 W/cm2 and 50% duty cycle with plasmid encoding enhanced green fluorescent protein (EGFP) as a report gene. Forty-eight h later, the expression of EGFP was detected under the fluorescence microscopy. Transfection efficacy was quantitatively assessed by flow cytometry, and cell viability was evaluated by trypan blue exclusion. The results showed that the transfection efficacy was increased with the increases in ultrasound output power and the ideal trans- fection efficacy was achieved in HepG2 cells irradiated by ultrasound at 0.8 W/cm2 for 30 s. The transfection efficacy in ulstrasound+P85 group was three times higher than in single ultrasound group [(17.63±1.07)% vs (5.57±0.56)%, P<0.051. The cell viability was about 81% and 62% in ultrasound group and ultrasound+P85 group respectively. It was concluded that ultrasound in combination with P85 could mediate the gene transfection of HepG2 cells, ideal transfection efficacy was achieved by ultrasound irradiation at 0.8 W/cm2 for 30 s, and P85 could somewhat increase the damage to cells caused by ultrasound.
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In order to assess whether gene transfection could be mediated by ultrasound in association with P85 and find the appropriate parameters of ultrasound irradiation, the effects of ultrasound with or without P85 on gene transfection of HepG2 cells were examined. The HepG2 cells were irradiated by ultrasound at 1 MHz, 0.4-2.0 W/cm(2) and 50% duty cycle with plasmid encoding enhanced green fluorescent protein (EGFP) as a report gene. Forty-eight h later, the expression of EGFP was detected under the fluorescence microscopy. Transfection efficacy was quantitatively assessed by flow cytometry, and cell viability was evaluated by trypan blue exclusion. The results showed that the transfection efficacy was increased with the increases in ultrasound output power and the ideal transfection efficacy was achieved in HepG2 cells irradiated by ultrasound at 0.8 W/cm(2) for 30 s. The transfection efficacy in ulstrasound+P85 group was three times higher than in single ultrasound group [(17.63+/-1.07)% vs (5.57+/-0.56)%, P<0.05]. The cell viability was about 81% and 62% in ultrasound group and ultrasound+P85 group respectively. It was concluded that ultrasound in combination with P85 could mediate the gene transfection of HepG2 cells, ideal transfection efficacy was achieved by ultrasound irradiation at 0.8 W/cm(2) for 30 s, and P85 could somewhat increase the damage to cells caused by ultrasound.
Subject(s)
Cell Survival/genetics , Green Fluorescent Proteins/genetics , Hep G2 Cells , Poloxalene/pharmacology , Transfection , UltrasonicsABSTRACT
To examine the role of ultrasound in gene delivery in vitro, three cells lines were exposed to the low-frequency ultrasound of varying intensities and for different durations to evaluate their effect on gene transfection and cell viability of the cells. Microbubble (MB), Optison (10%), was also used to observe the role of the microbubbles in gene transfection. The results demonstrated that as the ultrasound intensity and the exposure time increased, the gene transfer rate increased and the cell viability decreased, but at high energy intensities, the cell viability decreased dramatically, which caused the transfer rate to decrease. The most efficient ultrasound intensity for inducing gene transfer was 1 W/cm(2) with duration being 20 s. At the same energy intensity, higher ultrasound intensity could achieve maximal gene transfer rate earlier. Microbubbles could increase ultrasound-induced cell gene transfer rate by about 2 to 3 times mainly at lower energy intensities. Moreover, microbubbles could raise the maximum gene transfer rate mediated by ultrasound. It is concluded that the low-frequency ultrasound can induce cell gene transfer and the cell gene transfer rate and viability are correlated with not only the ultrasound energy intensity but also the ultrasound intensity, the higher ultrasound intensity achieves its maximal transfer rate more quickly and the ultrasound intensity that can induce optimal gene transfer is 1 W/cm(2) with duration being 20 s, and microbubbles can significantly increase the maximal gene transfer rate in vitro.