Near-Infrared Fluorescence Imaging Contrast Agents for Clinical Research: Limitations and Alternatives

Introduction: Near-infrared fluorescence imaging is a technique that will establish itself in the short term at the international level because it is recognized for its potential to improve the performance of surgical interventions, its moderate investment and operating costs and its portability. Although the technology is now mature, there is currently the problem of the availability of contrast agents to be injected IV. The aim of this methodology article is to propose an alternative solution to the need for contrast agents for clinical research, particularly in oncology. Methodology: They consist of coupling a fluorescent marker in the form of an NHS derivative, such as IR DYE manufactured in compliance with GMP, with therapeutic monoclonal antibodies having marketing authorization for molecular imaging. For a given antibody, the marking procedure must be the subject of a validation file on the final preparation filtered on a sterilizing membrane at 0.22 μm. Once the procedure has been validated, it would be unnecessary to repeat the tests before each clinical research examination. A check of the marking by thin-layer chromatography (TLC) and place it in a sample bank at +4˚C for 1 month of each injected formulation would be sufficient for additional tests if necessary. Conclusion: Molecular near-infrared fluorescence imaging is experiencing development, the process of which could be accelerated by greater availability of clinical contrast agents. Alternative solutions are therefore necessary to promote clinical research in this area. These methods must be shared to make it easier for researchers.

Diagnosis of focal liver lesions with deep learning-based multichannel analysis of hepatocyte-specific contrast-enhanced magnetic resonance imaging

BACKGROUND The nature of input data is an essential factor when training neural networks.Research concerning magnetic resonance imaging(MRI)-based diagnosis of liver tumors using deep learning has been rapidly advancing.Still,evidence to support the utilization of multi-dimensional and multi-parametric image data is lacking.Due to higher information content,three-dimensional input should presumably result in higher classification precision.Also,the differentiation between focal liver lesions(FLLs)can only be plausible with simultaneous analysis of multisequence MRI images.AIM To compare diagnostic efficiency of two-dimensional(2D)and three-dimensional(3D)-densely connected convolutional neural networks(DenseNet)for FLLs on multi-sequence MRI.METHODS We retrospectively collected T2-weighted,gadoxetate disodium-enhanced arterial phase,portal venous phase,and hepatobiliary phase MRI scans from patients with focal nodular hyperplasia(FNH),hepatocellular carcinomas(HCC)or liver metastases(MET).Our search identified 71 FNH,69 HCC and 76 MET.After volume registration,the same three most representative axial slices from all sequences were combined into four-channel images to train the 2D-DenseNet264 network.Identical bounding boxes were selected on all scans and stacked into 4D volumes to train the 3D-DenseNet264 model.The test set consisted of 10-10-10 tumors.The performance of the models was compared using area under the receiver operating characteristic curve(AUROC),specificity,sensitivity,positive predictive values(PPV),negative predictive values(NPV),and f1 scores.RESULTS The average AUC value of the 2D model(0.98)was slightly higher than that of the 3D model(0.94).Mean PPV,sensitivity,NPV,specificity and f1 scores(0.94,0.93,0.97,0.97,and 0.93)of the 2D model were also superior to metrics of the 3D model(0.84,0.83,0.92,0.92,and 0.83).The classification metrics of FNH were 0.91,1.00,1.00,0.95,and 0.95 using the 2D and 0.90,0.90,0.95,0.95,and 0.90 using the 3D models.The 2D and 3D networks'performance in the diagnosis of HCC were 1.00,0.80,0.91,1.00,and 0.89 and 0.88,0.70,0.86,0.95,and 0.78,respectively;while the evaluation of MET lesions resulted in 0.91,1.00,1.00,0.95,and 0.95 and 0.75,0.90,0.94,0.85,and 0.82 using the 2D and 3D networks,respectively.CONCLUSION Both 2D and 3D-DenseNets can differentiate FNH,HCC and MET with good accuracy when trained on hepatocyte-specific contrast-enhanced multi-sequence MRI volumes.

Gastrointestinal contrast-enhanced ultrasonography for diagnosis and treatment of peptic ulcer in children

BACKGROUND The detection rate of peptic ulcer in children is improving,with development of diagnostic procedures.Gastroscopy is the gold standard for the diagnosis of peptic ulcer,but it is an invasive procedure.Gastrointestinal contrast-enhanced ultrasonography(CEUS)has the advantages of being painless,noninvasive,nonradioactive,easy to use,and safe.AIM To investigate the clinical value of CEUS for diagnosis and treatment of peptic ulcer in children.METHODS We investigated 43 children with digestive tract symptoms in our hospital from January 2021 to June 2022.All children were examined by routine ultrasound,gastrointestinal CEUS,and gastroscopy.The pathological results of gastroscopy were taken as the gold standard.Routine ultrasonography was performed before gastrointestinal CEUS.Conventional ultrasound showed the thickness of the gastroduodenal wall,gastric peristalsis,and the adjacent organs and tissues around the abdominal cavity.Gastrointestinal CEUS recorded the thickness of the gastroduodenal wall;the size,location and shape of the ulcer;gastric peristalsis;and adjacent organs and tissues around the abdominal cavity.The results of routine ultrasound and gastrointestinal ultrasound were compared with those of gastroscopy to evaluate the diagnostic results and coincidence rate of routine ultrasound and gastrointestinal CEUS.All children received informed consent from their guardians for CEUS.This study was reviewed and approved by the hospital medical ethics committee.RESULTS Among the 43 children,17(15 male,2 female)were diagnosed with peptic ulcer by gastroscopy.There were 26 children with nonpeptic ulcer.There were eight cases of peptic ulcer and 35 of nonpeptic ulcer diagnosed by conventional ultrasound.The diagnostic coincidence rate of peptic ulcer in children diagnosed by conventional ultrasound was 79.1%(34/43),which was significantly different from that of gastroscopy(P=0.033).It indicates that the coincidence rate of gastrointestinal contrast-enhanced ultrasound and gastroscope is low.Fifteen cases of peptic ulcer and 28 of nonpeptic ulcer were diagnosed by CEUS.The diagnostic coincidence rate of peptic ulcer in children was 95.3%(41/43).There was no significant difference between CEUS and gastroscopy(P=0.655).It indicates that the coincidence rate of gastrointestinal contrast-enhanced ultrasound and gastroscope is high.CONCLUSION Gastrointestinal CEUS has a high coincidence rate in the diagnosis of peptic ulcer in children,and can be used as a preliminary examination method.

Nano/microparticles and ultrasound contrast agents

Microbubbles have been used for many years now in clinical practice as contrast agents in ultrasound imaging.Recently,their therapeutic applications have also attracted more attention.However,the short circulation time(minutes)and relatively large size(two to ten micrometers)of currently used commercial microbubbles do not allow effective extravasation into tumor tissue,preventing efficient tumor targeting.Fortunately,more multifunctional and theranostic nanoparticles with some special advantages over the traditional microbubbles have been widely investigated and explored for biomedical applications.The way to synthesize an ideal ultrasound contrast agent based on nanoparticles in order to achieve an expected effect on contrast imaging is a key technique.Currently a number of nanomaterials,including liposomes,polymers,micelles,dendrimers,emulsions,quantum dots,solid nanoparticles etc.,have already been applied to pre or clinical trials.Multifunctional and theranostic nanoparticles with some special advantages,such as the tumor-targeted(passive or active),multi-mode contrast agents(magnetic resonance imaging,ultrasonography or fluorescence),carrier or enhancer of drug delivery,and combined chemo or thermal therapy etc.,are rapidly gaining popularity and have shown a promising application in the field of cancer treatment.In this mini review,the trends and the advances of multifunctional and theranostic nanoparticles are briefly discussed.

Correlation study of biological characteristics of non-small cell lung cancer A549 cells after transfecting plasmid by microbubble ultrasound contrast agent

Objective: To explore the role of the abnormal expression of miRNAs in the development process of non-small cell lung cancer and the feasibility of ultrasound microbubble-mediated gene therapy after transfecting antisense miRNA-224 and miRNA-122 a plasmids into nonsmall cell lung cancer A549 cells. Methods: Antisense miRNA-224 and miRNA-122 a plasmids were transfected into non-small cell lung cancer A549 cells on the optimal ultrasound microbubble mediated condition. We set up a control group. The cell proliferation activity, apoptosis, invasion ability were detected by MTT assay, Annexin V-PE, Transwell invasion experiment and colony formation assay, respectively. Results: The expression of mi RNA-224 decreased and the expression of miRNA-122 a rose after the plasmids of target genes were transfected into non-small cell lung cancer A549 cells, and there were significant differences when compared with those of the control group(P<0.05). After the plasmids of target genes were transfected into A549 cells, the growth of antisense miRNA-224 and miRNA-122 a were inhibited, and the differences were significant as compared with the control group(P < 0.05). Besides, the inhibition of miRNA-122 a group was the most significant and there was statistically significant difference as compared with miRNA-224 group(t =-4.694, P = 0.009). After the plasmids of target genes were transfected into A549 cells, the proportion of apoptotic cells increased, the invasive cells were decreased and the clone ability reduced, and also there was a significant difference as compared with those of the control group(P < 0.05). What's more, the apoptotic peak appeared in miRNA-122 a group. Its invasion ability decreased most obviously(40.25 ± 3.97/visual field), the number of clone ability was 104.93 ± 4.87 and the inhibitory effect was the most obviously. There was statistically significant difference as compared with other groups(P < 0.05). Conclusions: A549 cells transfected by ultrasound microbubble-mediated antisense miRNA-224 and mi RNA-122 a plasmids possessed good transfection efficiency. The cell growth, invasion and colony forming abilities of transfected A549 cells were suppressed, which laid a solid foundation for the gene therapy of non-small cell lung cancer.

Application of Low Tube Voltage, Low-concentration Contrast Agent Using a 320-row CT in Coronary CT Angiography: Evaluation of Image Quality, Radiation Dose and Iodine Intake

The effect of low voltage and low concentration contrast agent on image quality of coronary CT angiography,radiation dose and iodine intake was evaluated.A total of 121 patients with body mass index(BMI)<26 kg/m2 and heart rate(HR)<70 beats/min were randomly divided into four groups:group A(n=31,80 kVp,270 mgl/mL);group B(n=33,100 kVp,270 mgl/mL);group C(h=30,100 kVp,320 mgl/mL);group D(w=27,100 kVp,400 mgl/mL).The automatic current modulation system and the iterative algorithm for reconstruction were adopted in each group.The CT values and SD values of the aortic root(AR),subcutaneous fat,left coronary artery opening(LCA),and right coronary artery opening(RCA)were measured in all groups,the signalto-noise ratio(SNR)and contrast noise ratio(CNR)were calculated,and effective radiation dose and iodine intake were recorded.The subjective assessment for image quality was performed by two physicians using a 4-point scale.The results were compared using the one-way ANOVA and rank sum tests.The image quality of the four groups met the clinical diagnostic requirements.The CT values of AR in groups A,B,C,and D were 537.6±71.4,447.2±81.9,445.2±64.9 and 518.5±94.9 Hu,respectively,with no significant difference between group A and group D,or between group B and group C,while CT values in groups B and C were significantly lower than those in groups A and D(P<0.05).In groups A,B,C,and D,the LCA SNR values were 22.7±9.1,23.3±9.1,23.3±7.7 and 26.6±8.9,and the RCA CNR values were 26.9±9.&28.5±11.4,27.7土&8 and 32」±10.6,respectively.The AR visual scores in groups A,B,C and D were 3.8±0.2,3.9±0.3,3.9±0.3 and 4.0±0.3,respectively.There were no significant differences in SNR,CNR and visual score among the four groups(P>0.05).The radiation doses in groups A,B,C and D were 2.6±1.4,3.6±1.&4.9±3.5 and 4.9±2.8 mSv,respectively.The radiation dose in group A was significantly less than that in the rest three groups(P<0.05).The iodine intakes in groups A,B,C and D were 14.9±1.5,15.0±1.5,17.7±2.0 and 18.1±2.5 g,respectively.There was no significant difference in the intake of iodine between groups C and D,or between groups A and B,while iodine intake in groups A and B were significantly reduced as compared with that in groups C and D(P<0.05).It was concluded that for patients with low BMI and controlled HR,compared to 100 kVp tube voltage combined with multiple concentration contrast agents,80 kVp combined with 270 mgl/mL contrast agent is enough to ensure the quality of the images,and can reduce the radiation dose significantly,while reducing the amount of iodine intake notably,thus reducing the incidence of adverse reaction.

Self-assembled superparamagnetic nanoparticles as MRI contrast agents — A review

Recent progress of the preparation and applications of superparamagnetic iron oxide（SPIO） clusters as magnetic resonance imaging（MRI） probes is reviewed with regard to their applications in labeling and tracking cells in vivo, in diagnosis of cardiovascular diseases and tumors, and in drug delivery systems. Magnetic nanoparticles（NPs）, especially SPIO nanoparticles, have long been used as MRI contrast agents and as an advantageous nanoplatform for drug delivery,taking advantage of their unique magnetic properties and ability to function at the molecular and cellular levels. Due to advances in nanotechnology, various means to control SPIO NPs＇ size, composition, magnetization and relaxivity have been developed, as well as ways to usefully modify their surface. Recently, self-assembly of SPIO NP clusters in particulate carriers — such as polymeric micelles, vesicles, liposomes, and layer-by-layer（Lb L） capsules — have been widely studied for application as ultrasensitive MRI probes, owing to their remarkably high spin–spin（T2） relaxivity and convenience for further functionalization.

A contrast study on different gasifying agents of underground coal gasification at Huating Coal Mine

To optimize the technological parameter of underground coal gasification （UCG）, the experimental results of air gasification, air-steam gasification, oxygen-enrichment steam gasification, pure oxygen steam gasification and two-stage gasification were studied contrastively based on field trial at the Huating UCG project. The results indicate that the average low heat value of gas from air experiment is the lowest （4.1 MJ/Nm3） and the water gas from two-stage gasification experiment is the highest （10.72 MJ/Nm3）. The gas productivity of air gasification is the highest and the pure oxygen steam gasification is the lowest. The gasification efficiency of air gasification, air-steam gasification, oxygen-enriched steam gasification, pure oxygen steam gasification and two-stage gasification is listed in ascending order, ranging from 69.88% to 84.81%. Described a contract study on results of a field test using steam and various levels of oxygen enrichment of 21%, 32%, 42% and 100%. The results show that, with the increasing of O2 content in gasifying agents, the gas caloricity rises, and the optimal O2 concentration range to increase the gas caloricity is 30%-40%. Meanwhile, the consumption of O2 and steam increase, and the air consumption and steam decomposition efficiency fall.

FEASIBILITY STUDY OF AN ULTRASOUND CONTRAST AGENT(LEVOVIST) IN COLOR DOPPLER IMAGING OF LIVER NEOPLASMS

The purpose of this study was to determine the efficacy of using an ultrasound contrast agent(levovist)to enhance the color Doppler imaging of liver neoplasms.Thirty patients with hepatic tumors were enrolled in this study.After intravenous administration of levovist,the color Doppler signals of normal hepatic vessels were enhanced.In various hepatic tumors,the different patterns of tumor vascularity were observed,which had not been demonstrated in conventional non contrast color Doppler imaging.In 11 of 16 patients with hepatocarcinoma,additional color Doppler signals were observed in the central part of the tumors.On the contrary,3 patients with metastatic liver lesions the enhanced color Doppler signals appear only at the peripheral of tumors.A typical rim like color enhancement was seen in 2 of the 3 cases.In six patients with hepatic hemangiomas contrast enhanced color Doppler imaging demonstrated the blood vessels at the margin of the neoplasms.Contrast enhanced color Doppler imaging improves the visualization of the hepatic neoplasm vascularity.This technique holds great promise for detecting small liver tumors and differentiating hepatic neoplasms.

Contrast-enhanced ultrasound using SonoVue mixed with oral gastrointestinal contrast agent to evaluate esophageal hiatal hernia: Report of three cases and a literature review

BACKGROUND Due to a thicker abdominal wall in some patients,ultrasound artifacts from gastrointestinal gas and surrounding tissues can interfere with routine ultrasound examination,precluding its ability to display or clearly show the structure of a hernial sac(HS)and thereby diminishing diagnostic performance for esophageal hiatal hernia(EHH).Contrast-enhanced ultrasound(CEUS)imaging using an oral agent mixture allows for clear and intuitive identification of an EHH sac and dynamic observation of esophageal reflux.CASE SUMMARY In this case series,we report three patients with clinically-suspected EHH,including two females and one male with an average age of 67.3±16.4 years.CEUS was administered with an oral agent mixture(microbubble-based SonoVue and gastrointestinal contrast agent)and identified a direct sign of supradiaphragmatic HS(containing the hyperechoic agent)and indirect signs[e.g.,widening of esophageal hiatus,hyperechoic mixture agent continuously or intermittently reflux flowing back and forth from the stomach into the supradiaphragmatic HS,and esophagus-gastric echo ring(i.e.,the“EG”ring)seen above the diaphragm].All three cases received a definitive diagnosis of EHH by esophageal manometry and gastroscopy.Two lesions resolved upon drug treatment and one required surgery.The recurrence rate in follow-up was 0%.The data from these cases suggest that the new non-invasive examination method may greatly improve the diagnosis of EHH.CONCLUSION CEUS with the oral agent mixture can facilitate clear and intuitive identification of HS and dynamic observation of esophageal reflux.

Non-thermal ablation of rabbit liver VX2 tumor by pulsed high intensity focused ultrasound with ultrasound contrast agent:Pathological characteristics

AIM： To investigate the pathological characteristics of non-thermal damage induced by pulsed high intensity focused ultrasound （PHIFU） combined with ultrasound contrast agent （UCA）, SonoVue （Bracco SpA, Milan, Italy） in rabbit liver VX2 tumor. METHODS： Liver VX2 tumor models were established in 20 rabbits, which were divided randomly into PHIFU combined with ultrasound contrast agent group （PHIFU ＋ UCA group） and sham group. In the PHIFU ＋ UCA group, 0.2 mL of SonoVue was injected intravenously into the tumor, followed by ultrasound exposure of Isp 5900 W/cm^2. The rabbits were sacrificed one day after ultrasound exposure. Specimens of the exposed tumor tissues were obtained and observed pathologically under light microscope and transmission electron microscope. The remaining tumor tissues were sent for 2,3,5-Triphenyltetrazolium chloride （TTC） staining. RESULTS： Before Trc staining, tumor tissues in both the sham and the PHIFU ＋ UCA groups resembled gray fish meat, After TIC staining, the tumor tissues were uniformly stained red, with a clear boundary between tumor tissue and normal tissue, Histological examination showed signs of tumor cell injury in PHIFU ＋ UCA group, with cytoplasmic vacuoles of various sizes, chromatin margination and karyopyknosis. Electron microscopic examination revealed tumor cell volume reduction, karyopyknosis, chromatin margination, intercellular space widening, the presence of high electro＇n-density apoptotic bodies and vacuoles in cytoplasm. CONCLUSION： The non-thermal effects of PHIFU combined with UCA can be used to ablate rabbit liver VX2 tumors.

Photoacoustic molecular imaging using combined acupuncture and gold nanorods as a composite contrast agent

In this study,we developed a novel photoacoustic imaging technique based on poly(ethyleneglycol)-coated(PEGylated)gold nanorods(PEG-GNRs)(as the contrast agent)combined with traditional Chinese medicine(TCM)acupuncture(as the auxiliary method)for quantitatively monitoring contrast enhancement in the vasculature of a mouse brain in vivo.This study takes advantage of the strong near-infrared absorption(peak at700 nm)of GNRs and the ability to adjust the hemodynamics of acupuncture.Experimental results show that photoacoustic tomography(PAT)successfully reveals the optical absorption variation of the vasculature of the mouse brain in response to intravenous administration of GNRs and acupuncture at the Zusanli acupoint(ST36)both individually and combined.The quantitative measurement of contrast enhancement indicates that the composite contrast agents(integration of acupuncture and GNRs)would greatly enhance the photoacoustic imaging contrast.The quantitative results also have the potential to estimate the local concentration of GNRs and even the real-time effects of acupuncture.

Gilbert syndrome combined with prolonged jaundice caused by contrast agent:Case report

This case highlights a patient with Gilbert syndrome who underwent endoscopic retrograde cholangiopancreatography(ERCP) with removal of bile duct stones, who then experienced an unexplained increase in bilirubin, with total bilirubin(TBIL) levels increasing from 159.5 μmol/L to 396.2 μmol/L and to a maximum of 502.8 μmol/L after 9 d. Following the decrease in the TBIL level, enhanced magnetic resonance cholangiopancreatography(MRCP) was performed to exclude any possible remaining choledocholithiasis. Nevertheless, the serum bilirubin level increased again, with TBIL levels rising from 455.7 μmol/L to 594.8 μmol/L and a maximum level of 660.3 μmol/L with no remaining bile duct stones. A liver biopsy showed severe bile duct cholestasis with no inflammation. Based on the exclusion of other potential causes of hyperbilirubinemia and the fact that both instances of increased bilirubin occurred after ERCP and MRCP, the contrast agents iopromide and gadoterate meglumine were suspected to be the causes of the hyperbilirubinemia. As of the writing of this report, the patient's bilirubin levels have spontaneously returned to baseline levels. In summary,ERCP and MRCP utilizing the contrast agents iopromide and gadoterate meglumine may possibly induce prolonged hyperbilirubinemia.

Optimized energy thresholds in a spectral computed tomography scan for contrast agent imaging

Spectral computed tomography(CT) based on photon counting detectors(PCDs) is a well-researched topic in the field of X-ray imaging. When PCD is applied in a spectral CT system, the PCD energy thresholds must be carefully selected, especially for K-edge imaging, which is an important spectral CT application. This paper presents a threshold selection method that yields better-quality images in K-edge imaging. The main idea is to optimize the energy thresholds ray-by-ray according to the targeted component coefficients, followed by obtaining an overall optimal energy threshold by frequency voting. A low-dose pre-scan is used in practical implementations to estimate the line integrals of the component coefficients for the basis functions. The variance of the decomposed component coefficients is then minimized using the Cramer–Rao lower bound method with respect to the energy thresholds. The optimal energy thresholds are then used to take a full scan and gain better image reconstruction with less noise than would be given by a full scan using the non-optimal energy thresholds. Simulations and practical experiments on imaging iodine and gadolinium solutions, which are commonly used as contrast agents in medical applications, were used to validate the method. The noise was significantly reduced with the same dose relative to the non-optimal energy thresholds in both simulations and in practical experiments.

Oleic Acid (OA), A Potential Dual Contrast Agent for Postmortem MR Angiography (PMMRA): A Pilot Study

Summary:Choosing proper perfusates as contrast agents is an important aspect for postmortem magnetic resonance angiography(PMMRA).However,in this emerging field,the number of suitable kinds of liquid is still very limited.The objective of this research is to compare MR images of oleic acid(OA)with paraffin oil(PO)in vitro and in ex situ animal hearts,in order to evaluate the feasibility to use OA as a novel contrast agent for PMMRA.In vitro,OA,PO and water(control)were introduced into three tubes separately and T,weighted-spin echo(Tw SE)and T2w-SE images were acquired on a 1.5T MR scanner.In the second experiment,0A and PO were injected into left coronary artery(LCA)and left ventricle(LV)of ex situ bovine hearts and their Tw-SE,Tzw-SE,Tw-multipoint Dixon(Tjw-mDixon)and 3DT2w-mDixon images were acquired.The overall results indicate that OA may have a potential to be used as a dual(T and T2 based)contrast agent for PMMRA when proper sequence parameters are utilized.However,as the pilot study was based on limited number of animal hearts,more researches using OA in cadavers are needed to validate our findings.

Role of microbubble ultrasound contrast agents in the non-invasive assessment of chronic hepatitis C-related liver disease

Patients who are chronically infected with the hepatitis C virus often develop chronic liver disease and assessment of the severity of liver injury is required prior to considering viral eradication therapy. This article examines the various assessment methods currently available from gold standard liver biopsy to serological markers and imaging. Ultrasound is one of the most widely used imaging modalities in clinical practice and is already a first-line diagnostic tool for liver disease. Microbubble ultrasound contrast agents allow higher resolution images to be obtained and functional assessments of microvascular change to be carried out. The role of these agents in quantifying the state of hepatic injury is discussed as a viable method of determining the stage and grade of liver disease in patients with hepatitis C. Although currently confined to specialist centres, the availability of microbubble contrast-enhanced ultrasound will inevitably increase in the clinical setting.

Study on Multi-density Contrast Agent Fillers of Duct Casting Based on CT Three-Dimensional Reconstruction

The three-dimensional visualization model of human body duct is based on virtual anatomical structure reconstruction with duct angiography,which realizes virtual model transferred from two-dimensional,planar and static images into three-dimensional,stereoscopic and dynamic ones repectively.In recent years,the multi-duct segmentation and division of the same specimen（or organ） is the focus of attention shared by surgeons and clinical anatomists.On the basis of 4.22 g/cm3 body bone density,this study has screened out metal oxide contract agent with different density for infusion and modeling,as well as compared and analyzed the effects of three-dimensional image of CT virtual bronchoscopy（CTVB）,three-dimensional image of CT maximum intensity projection and three-dimensional model.This experiment result showed synchronously infusing multi-duct of same specimen（or organ） with contrast agent in different densities could reconstruct three-dimensional models of all ducts once only and adjust threshold to develop single or multiple ducts.It was easier to segment and observe the duct structure,anastomosis,directions and crossing in different parts,which was beyond comparison with three-dimensional image of CTVB.Although the existing three-dimensional duct reconstruction techniques still cannot be applied in living bodies temporarily,this study focused on a creative design of ducts segmentation in different density,which proposed a new experimental idea for developing multi-duct three-dimensional model in living body in the future.It will play a significant role in disease diagnosis and individual design in surgical treatment program.Therefore,this study observes the three-dimensional status of human duct with the application of contrast agent fillers in different density,combined with three-dimensional reconstruction technology.It provides an innovative idea and method for constructing three-dimensional model of digital multi-duct specimen,and the ultimate goal is to develop the digitized virtual human and precise medical treatment better and faster.

PREPARATION OF POLYELECTROLYTE MULTILAYER COATED MICROBUBBLES FOR USE AS ULTRASOUND CONTRAST AGENT

Objeelive To prepare and characterize polyelectrolyte multilayer film coated microbubbles for use as ultrasound contrast agent （UCA） and evaluate its effects in ultrasonic imaging on normal rabbit＇s fiver parenchyma. Methods Perfluorocarbon （PFC）-containing microbubbles （ST68-PFC） were prepared by sonication based on suffactant （ Span 60 and Tween 80）. Subsequently, the resulting ST68-PFC microbubbles were coated using oppositely charged polyelectrolytes by microbubble-templated layer-by-layer self-assembly technique via electrostatic interaction. The enhancement effects in ultrasonic imaging on normal rabbit＇s liver parenchyma were assessed. Results The obtained microbubbles exhibited a narrow size distribution. The polyelectrolytes were successfully assembled onto the surface of ST68-PFC microbubbles. In vivo experiment showed that polyelectrolyte multilayer film coated UCA effectively enhanced the imaging of rabbit＇s liver parenchyma. Conclusions The novel microbubbles UCA coated with polyelectrolyte multilayer, when enabled more function, has no obvious difference in enhancement effects compared with the pre-modified microbubbles. The polymers with chemically active groups （ such as amino group and carboxyl group） can be used as the outermost layer for attachment of targeting ligands onto microbubbles, allowing selective targeting of the microbubbles to combine with desired sites.

Lactoferrin-Conjugated Polylactic Acid Nanobubbles Encapsulated Perfluoropentane as a Contrast Agent for Ultrasound/Magnetic Resonance Dual-Modality Imaging

The development of contrast agents that can be activated by multiple modes is of great significance for tumor diagnosis.In this study,the lactoferrin(Lf)-conjugated polylactic acid(PLLA)nanobubbles(Lf-PLLA NBs)were used to encapsulate liquid perfluoropentane(PFP)with the double emulsion method,creating PFP loaded(PFP/Lf-PLLA)NBs for the ultrasound/magnetic resonance dual-modality imaging of subcutaneous tumor.The parti-cle diameter and stability of nanobubbles were investigated by photon correlation spectroscopy.The biocompat-ibility of nanobubbles was preliminarily evaluated by cell proliferation and migration assay,hemolysis rate,and blood biochemistry analysis.A B-mode clinical ultrasound real-time imaging system was used to perform ultra-sonic imaging in vivo.Magnetic resonance imaging in vivo was applied with a clinical 3.0 T magnetic resonance imaging(MRI)scanner system.The mean particle diameter of PFP/Lf-PLLA NBs was 320.2±4.1 nm with a low polydispersity index(PDI,0.145±0.025),and the NBs were negatively charged(−11.4±0.4 mV).The transmis-sion electron microscopy(TEM)results showed that PFP/Lf-PLLA NBs exhibited highly monodispersed and pos-sessed an obvious spherical structure of nanocapsules.Nanobubbles had good stability at 4°C.Different concentrations of the PFP/Lf-PLLA NBs solution had no effect on the cell in cytotoxicity and cell migration,and the results of hemolysis rate and blood biochemistry assay also indicated the good biocompatibility of NBs.On the ultrasound/magnetic resonance imaging of tumor-bearing mice,PFP/Lf-PLLA NBs showed signifi-cantly enhanced contrast ability of tumor tissue.Therefore,PFP/Lf-PLLA NBs had great potential to be a contrast agent for tumor dual-modality imaging in vivo.

Dynamics of an ultrasound contrast agent microbubble near spherical boundary in ultrasound field

The goal of this article is to establish the conditions of excitation where one has to deal with ultrasound contrast agent(UCA) microbubbles pulsating near biological tissues with spherical boundary in ultrasound field for targeted drug delivery and cavitation-enhanced thrombolysis, etc., and contributes to understanding of mechanisms at play in such an interaction. A modified model is presented for describing microbubble dynamics near a spherical boundary(including convex boundary and concave boundary) with an arbitrary-sized aperture angle. The novelty of the model is such that an oscillating microbubble is influenced by an additional pressure produced by the sound reflection from the boundary wall. It is found that the amplitude of microbubble oscillation is positively correlated to the curve radius of the wall and negatively correlated to the aperture angle of the wall and the sound reflection coefficient. Moreover, the natural frequency of the microbubble oscillation for such a compliable wall increases with the wall compliance, but decreases with the reduction of the wall size, indicating distinct increase of the natural frequency compared to a common rigid wall. The proposed model may allow obtaining accurate information on the radiation force and signals that may be used to advantage in related as drug delivery and contrast agent imaging.