OBJECTIVETo observe the cell binding characteristics of SonoVue microbubbles targeting choriocarcinoma cells and provide evidence for clinical ultrasonic localization of the tumor utilizing the microbubbles.

METHODSThe targeted microbubbles were prepared by conjugating anti-HCG antibody with the SonoVue microbubbles and added in choriocarcinoma cells or endometrial stromal cells to compare the cell binding rate of the agents under optical microscope and with flow cytometry.

RESULTSFlow cytometry demonstrated a binding rate of 77.6% between the SonoVue microbubbles and anti-HCG antibody. Light microscopy showed that the total rosette formation rate of the choriocarcinoma cells exposed to the targeted microbubble bearing anti-HCG antibody reached (87.8-/+6.3)%, significantly higher than that of the endometrial stromal cells [(9.4-/+1.7)%, P<0.05]. The binding rate of the targeted microbubbles with the choriocarcinoma cells before and after PBS washing were (85.4-/+4.7)% and (83.1-/+3.8)% (P>0.05), respectively, suggesting strong stability of the binding. The binding rate was 81.0% according to the results of flow cytometry.

CONCLUSIONThe targeted microbubbles as a contrast agent can efficiently bind to the choriocarcinoma cells in vitro with a stability sufficient to resist the blood flow.

Choriocarcinoma ; pathology ; Contrast Media ; metabolism ; Female ; Humans ; Microbubbles ; Phospholipids ; metabolism ; Pregnancy ; Sulfur Hexafluoride ; metabolism ; Uterine Neoplasms ; metabolism

OBJECTIVETo investigate the distribution of hyaluronic acid (HA) with iohexol tracing in the knee joint cavity of rabbits using CT plain scan, three-dimensional reconstruction and Χ-ray and observe how different injection sites affect HA distribution.

METHODSMixtures of HA and iohexol (tracer) were prepared that contained final iohexol concentrations of 2.5%, 5%, 10%, 20%, or 40%. The HA-iohexol mixtures (0.5 ml) were injected into rabbit knee joints, and the optimal iohexol concentration that allowed clear differentiation of the injected agents from the surrounding tissues was determined using dual-source CT plain scan and three-dimensional reconstruction technique. The HA-iohexol mixture (0.5 ml) containing the optimal concentration of iohexol was then injected into the knees of the rabbits either through the patella medial approach or the medial joint line approach, and HA distribution in the knee joint cavity was observed using CT scan and Χ-ray.

RESULTSThe CT value of HA-iohexol mixture increased progressively with the tracer concentration. After injection of the mixture containing 2.5%, 5%, 10%, 20%, and 40% iohexol, the CT value ratios of the soft tissue, HA-iohexol mixture and bone cortex were 2:7:46, 2:14:44, 2:28:44, 2:60:46, and 2:98:45, respectively, and a iohexol concentration of 5% was determined as optimal for differntiating the injected agents from the surrounding tissues. The HA-iohexol mixutre containing 5% iohexol injected through the medial-patellar approach was distributed mainly over the patello-femoral joint, and that injected through the joint line approach was found mainly over the tibio-femoral joint.

CONCLUSIONHA-iohexol mixture containing 5% iohexol allows clear differentiation of bone cortex and soft tissues in rabit knee joint from the injected agents on CT scan and Χ-ray, and the injection approach can influence HA distribution in the knee joint cavity.

Animals ; Contrast Media ; Hyaluronic Acid ; metabolism ; Iohexol ; Knee Joint ; Rabbits ; Tissue Distribution ; Tomography, X-Ray Computed

BACKGROUNDThe cellular plasma membrane represents a natural barrier to many exogenous molecules including magnetic resonance (MR) contrast agent. Cell penetrating peptide (CPP) is used to internalize proteins, peptides, and radionuclide. This study was undertaken to assess the value of a new intracellular MR contrast medium, CPP labeled diethylenetriamine pentaacetic acid gadolinium (Gd-DTPA) in molecular imaging in vitro.

METHODSFluorescein-5-isothiocyanate (FITC) and Gd-DTPA respectively labeled with CPP (FITC-CPP, Gd-DTPA-CPP) were synthesized by the solid-phase method. Human hepatic cancer cell line-HepG2 was respectively stained by FITC-CPP and FITC to observe the uptake and intracellular distribution. HepG2 was respectively incubated with 100 nmol/ml Gd-DTPA-CPP for 0, 10, 30, 60 minutes, and imaged by MR for studying the relationship between the incubation time and T(1)WI signal. The cytotoxicity to NIH3T3 fibroblasts cells was measured by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide reduction assay (MTT).

RESULTSThe molecular weights of CPP labeled imaging agents, which were determined by MALDI mass spectrometry (FITC-CPP MW = 2163.34, Gd-DTPA-CPP MW = 2285.99), were similar to the calculated molecular weights. Confocal microscopy suggested HepG2 translocated FITC-CPP in cytoplasm and nucleus independent with the incubation temperature. MR images showed HepG2 uptaken Gd-DTPA-CPP had a higher T(1) weighted imaging (T(1)WI) signal, and that the T(1)WI signal intensity was increasing in a time-dependent manner (r = 0.972, P = 0.001), while the signal intensity between the cells incubated by Gd-DTPA for 60 minutes and the controlled cells was not significantly different (P = 0.225). By MTT, all concentrations from 50 nmol/ml to 200 nmol/ml had no significant (F = 0.006, P = 1.000) effect on cell viability of mouse NIH3T3 fibroblasts, compared with the control group.

CONCLUSIONSThe newly constructed CPP based on polyarginine can translocate cells by carrying FITC and MR contrast agent Gd-DTPA, and the intracellular concentrations are readily detectable by MR imaging, suggesting a new way for MR molecular imaging.

Cell Line, Tumor ; Cell Membrane Permeability ; Contrast Media ; Fluorescein-5-isothiocyanate ; Gadolinium DTPA ; Humans ; Magnetic Resonance Imaging ; methods ; Peptides ; metabolism

OBJECTIVETo assess the binding ability of microbubbles targeted to alphavbeta3-integrin (MBp) for thrombus-targeted contrast-enhanced ultrasound.

METHODSTargeted microbubbles were prepared by conjugating the monoclonal antibody against alphavbeta3-integrin to lipid shell of the microbubble via the avidin-biotin bridges. Equivalent isotype control microbubbles (MB) or targeted ultrasound microbubbles (MBp) were randomly added into the flow chamber. After a 30-min incubation with the thrombus fixed in an agarose flow chamber model, the thrombus was washed with a continuous flow of PBS solution (15 cm/s) for 2, 4, 6, 8 and 10 min, followed by thrombus imaging using contrast-enhanced ultrasound and measurement of the video intensity (VI) values of the images.

RESULTSThe VI of the thrombus in MBp group was reduced by 28%-66%, while that in control MB group was decreased by 87%-94%, and the VI values of the thrombus group were significantly greater in former group at each of the time points (P<0.05).

CONCLUSIONMBP has good targeting ability to the thrombus with resistance to the shear stress after adhesion to the thrombus. In vitro evaluation of the thrombus-binding capability of the targeted microbubble (MBp) by simulating the shear stress in vivo can be helpful for predicting the in vivo effects of ultrasonic molecular imaging using MBp.

Antibodies, Monoclonal ; chemistry ; immunology ; Contrast Media ; chemistry ; Humans ; Integrin alphaVbeta3 ; immunology ; metabolism ; Microbubbles ; Sepharose ; Thrombosis ; diagnostic imaging ; Ultrasonography

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.
3T3 Cells ; CHO Cells ; Cell Line ; Cell Survival/*genetics ; Contrast Media/metabolism ; Cricetinae ; Cricetulus ; Microbubbles ; Transfection/*methods ; Ultrasonics

OBJECTIVE: To investigate the diagnostic significance of basophil activation test (BAT) in anaphylaxis to non-ionic contrast media through testing the content of CD63, mast cell-carboxypeptidase A3 (MC-CPA3), and terminal complement complex SC5b-9 of the individuals by testing their levels in the normal immune group and the anaphylaxis groups to β-lactam drugs and non -ionic contrast media. METHODS: The CD63 expression of basophilic granulocyte in blood was detected by flow cytometry. The levels of MC-CPA3 in blood serum and SC5b-9 in blood plasma were detected by ELISA. RESULTS: The CD63 expression of basophilic granulocyte in blood, the levels of MC-CPA3 and SC5b-9 of anaphylaxis to non-ionic contrast media and β-lactam drugs were significantly higher than that in normal immune group (P < 0.05). CONCLUSION There is activation of basophilic granulocytes, mast cells and complement system in anaphylaxis to non-ionic contrast media. BAT can be used to diagnose the anaphylaxis to non-ionic contrast media.
Anaphylaxis/diagnosis* ; Basophils/cytology* ; Carboxypeptidases A/metabolism* ; Complement Membrane Attack Complex/metabolism* ; Contrast Media ; Flow Cytometry ; Granulocytes/cytology* ; Humans ; Mast Cells/cytology* ; Tetraspanin 30/metabolism*

Splenic hamartoma is a very rare benign tumor, which is usually found incidentally after splenectomy or autopsy. Although percutaneous needle biopsy can be performed, it carries a high risk of bleeding after the procedure. Therefore, diagnosis is usually made by surgical resection. Herein, we report a case of splenic hamartoma diagnosed by magnetic resonance imaging and contrast-enhanced ultrasonography, which enables visualization of the unique signals of microbubbles in the vessels in real time. Relevant literature is also reviewed.
Adult ; Antigens, CD31/metabolism ; Antigens, CD34/metabolism ; Contrast Media ; Female ; Hamartoma/*diagnosis/pathology/ultrasonography ; Humans ; Immunohistochemistry ; Magnetic Resonance Imaging ; Splenic Diseases/*diagnosis/pathology/ultrasonography ; Tomography, X-Ray Computed

BACKGROUND AND OBJECTIVEPET/CT imaging is expensive, so searching the tumor imaging agent for SPECT/CT is necessary. 99Tc(m) -N(NOEt)2 [bis (N-ethoxy-N-ethyl dithiocarbamato) nitrido 99Tc(m) (V)] can be uptaken by lung cancer cells and other cells alike. The aim of this study is to evaluate the distinctive value in lung tumor with 99Tc(m) -N(NOEt)2, the difference in its uptake kinetics in human embryonic lung diploid fibroblasts KMB17 and several kinds of lung cancer cells lines.

METHODSFirstly, six different cell culture medium which contained YTMLC Gejiu human lung squamous carcinoma cell, SPC-A1 human lung adenocarcinoma cell, AGZY low metastatic human lung adenocarcinoma, 973 high metastatic human lung adenocarcinoma cell, GLC-82 Gejiu human lung adenocarcinoma cell, and KMB17 human embryonic lung diploid fibroblast, respectively with equal cell density of 1 x 10(6)/mL and the same volume were prepared; secondly, the same radioactive dose of 99Tc(m) -N(NOEt)2 was added into each sample and then 300 microL mixed sample was taken out respectively and cultured in 37 degrees C culture box; Finally, 5 min, 15 min, 30 min, 45 min, 60 min, 75 min, 90 min after cultivation, centrifuged each cultured sample and determined the intracellular radiocounts of each sample, calculated each cell sample's uptake rate of 99Tc(m) -N(NOEt)2 at different time.

RESULTSStatistical difference was found among six cell samples, and the uptake rate sequence from high to low is 973 and SPC-A1 > YTMLC > GLC-82 > AGZY > KMB17 respectively; furthermore, 30 min-45 min after culture, the uptake rate reached stability, and the 45 min uptake rate of each sample was higher than its 96.7% uptake peak.

CONCLUSIONBased on the results above mentioned, it is supposed that there are discriminative clinical value when using 99Tc(m) -N(NOEt)2 as a tumor targeting imaging agent, and 30 min or so after injection may be the best imaging time in the early imaging stage.

Cell Line ; Cell Line, Tumor ; Contrast Media ; pharmacokinetics ; Fibroblasts ; cytology ; metabolism ; Humans ; Lung Neoplasms ; metabolism ; Organotechnetium Compounds ; pharmacokinetics ; Tomography, Emission-Computed, Single-Photon

OBJECTIVETo analyze characteristic CT enhancement patterns of noncalcified pulmonary tuberculomas and their pathological basis.

METHODFifty-six patients with noncalcified pulmonary tuberculomas underwent surgical resection of the tuberculomas. Enhanced CT images of these tuberculomas were reviewed and analyzed in relation to the histological findings.

RESULTSOf the 56 patients, 45 showed no enhancement in the tuberculomas, which were histologically characterized by central caseous necrosis and a poorly vascularized peripheral fibrotic zone. Eleven patients showed ring-like or eggshell enhancement, and the central low density region was histologically confirmed to be caused by caseous or liquefied necrosis, while the ring enhancement resulted pathologically from moderately or well vascularized peripheral fibrotic or granulomatous tissues.

CONCLUSIONSPulmonary tuberculomas consists mainly of caseous necrotic tissues characterized by no enhancement and ring or eggshell enhancement on dynamic contrast-enhanced CT.

Adult ; Calcinosis ; Contrast Media ; Female ; Humans ; Male ; Middle Aged ; Tomography, X-Ray Computed ; Tuberculoma ; diagnostic imaging ; metabolism ; Tuberculosis, Pulmonary ; diagnostic imaging ; metabolism ; Young Adult

Primary liver carcinosarcoma is extremely rare. We report a case of liver carcinosarcoma in a 58-year-old man, which was identified by pathologic and immunohistochemical (IHC) examinations. Plain CT scans showed a hypodense mass in the right liver lobe adjacent to the gallbladder fossa. The triple-phase contrast CT scans showed a mixed density mass with inhomogeneous enhancement. Multiple cystic nodules with irregular rim enhancement of the margin located in the tumor. The gallbladder wall and transverse colon were involved. CT presentations of liver carcinosarcoma were unspecific and the pre-operative diagnosis is difficult.
Actins ; metabolism ; Antigens, Neoplasm ; metabolism ; Carcinosarcoma ; diagnostic imaging ; metabolism ; pathology ; Cell Adhesion Molecules ; metabolism ; Contrast Media ; Epithelial Cell Adhesion Molecule ; Humans ; Keratins ; metabolism ; Liver Neoplasms ; diagnostic imaging ; metabolism ; pathology ; Male ; Middle Aged ; Tomography, X-Ray Computed