Miao medicine Gu Yan Xiao tincture inhibits mTOR to stimulate chondrocyte autophagy in a rabbit model of osteoarthritis.

ETHNOPHARMACOLOGICAL RELEVANCE: The Gu Yan Xiao tincture, a blend of traditional Chinese herbs, is traditionally used for osteoarthritis and related pain. This study investigated its mechanism of action in order to rationalize and validate its therapeutic use. AIM OF THE STUDY: This study analyzed, in a rabbit model of knee osteoarthritis, whether and how Gu Yan Xiao tincture exerts therapeutic benefits by modulating chondrocyte autophagy. MATERIALS AND METHODS: The active constituents within the GYX tincture were identified using liquid chromatography-mass spectrometry. The rabbit model was established by injecting animals with type II collagenase intra-articularly, and the effects of topically applied tincture were examined on osteoarthritis lesions of the knee using histopathology, micro-computed tomography and x-ray imaging. Effects of the tincture were also evaluated on levels of inflammatory cytokines, matrix metalloproteases, and autophagy in chondrocytes. As a positive control, animals were treated with sodium diclofenac. RESULTS: The tincture mitigated the reduction in joint space, hyperplasia of the synovium and matrix metalloproteases in serum that occurred after injection of type II collagenase in rabbits. These therapeutic effects were associated with inhibition of mTOR and activation of autophagy in articular chondrocytes. Inhibiting mTOR with rapamycin potentiated the therapeutic effects of the tincture, while inhibiting autophagy with 3-methyladenine antagonized them. CONCLUSIONS: Gu Yan Xiao tincture mitigates tissue injury in a rabbit model of osteoarthritis, at least in part by inhibiting mTOR and thereby promoting autophagy in chondrocytes. These results rationalize the use of the tincture not only against osteoarthritis but also potentially other diseases involving inhibition of autophagy in bones and joints.

ALG3-CDG: lethal phenotype and novel variants in Chinese siblings.

Congenital disorders of glycosylation (CDG) are a group of genetic, mostly multisystem disorders, which often involve the central nervous system. ALG3-CDG is one the some 130 known CDG. Here we report two siblings with a severe phenotype and intrauterine death. Whole-exome sequencing revealed two novel variants in ALG3: NM_005787.6:c.512G>T (p.Arg171Leu) inherited from the mother and NM_005787.6:c.511C>T (p.Arg171Trp) inherited from the father.

Fatigue Performance of the CA Mortar Used in CRTS I Ballastless Slab Track under Simulated Servicing Condition.

The cement and asphalt mortar (CA mortar) used in the China Railway Track System (CRTS) I ballastless slab track may encounter a coupling fatigue effect under the high-frequency vibration, load and high-and-low temperature cycles, and the deterioration under fatigue may happen during service of the high-speed railway. In this study, the performance degradation and its mechanism of the CA mortar with and without polymer emulsion incorporated under the coupling fatigue effects of the high-frequency vibration, load and temperature were studied by using an anti-fatigue testing device specially developed for the CA mortar used in the ballastless slab track of the high-speed railway. The results showed that the deformation capacity of the CA mortar for CRTS I slab ballastless slab track decreased after fatigue test under simulated service environment, presenting a typical brittle characteristic and an obvious reduction of the ductility and toughness. The Scanning Electron Microscopy (SEM) observation and the mercury intrusion porosimetry (MIP) analysis showed that the volume of the macropore decreased whereas that of the micropore increased after the fatigue test. The asphalt in the hardened CA mortar revealed a softening and migration from the bulk paste to fill the pore and make the structure denser and even ooze out of the CA mortar under the high-frequency vibration and high temperature. Through incorporating the polymer emulsion, the anti-fatigue property of the CA mortar was obviously improved, which can prevent the CA mortar from losing its elastic adjustment function too early. Though increase of the strength and elastic modulus for the CA mortar after severe service is beneficial to the stability of train running, the comfort level and safety of the train operation may decline due to the gradual reduction of the ductility & toughness and the gradual loss of the elastic damping adjustment function of the CA mortar between the base concrete slab and the track slab.

Henoch-Schönlein Purpura With Testicular Necrosis: Sonographic Findings at the Onset, During Treatment, and at Follow-up.

Henoch-Schönlein purpura (HSP) with involvement of the testes is extremely rare. Here, we present a pediatric case of HSP involving testicular ischemic necrosis in an 8-year-old child. Ultrasonography plays an important role not only in the differential diagnosis at onset, but also in the assessment of treatment response and prognosis during treatment and at follow-up. In this case report, we present the sonographic images for the entire course of testicular involvement in HSP and reveal histopathologically the pathogeny as testicular autoimmune vasculitis caused by the deposition of immunoglobulin A-containing immune complexes in the testicular vessels.

CO2 bubbling-based 'Nanobomb' System for Targetedly Suppressing Panc-1 Pancreatic Tumor via Low Intensity Ultrasound-activated Inertial Cavitation.

Noninvasive and targeted physical treatment is still desirable especially for those cancerous patients. Herein, we develop a new physical treatment protocol by employing CO2 bubbling-based 'nanobomb' system consisting of low-intensity ultrasound (1.0 W/cm(2)) and a well-constructed pH/temperature dual-responsive CO2 release system. Depending on the temperature elevation caused by exogenous low-intensity therapeutic ultrasound irradiation and the low pH caused by the endogenous acidic-environment around/within tumor, dual-responsive CO2 release system can quickly release CO2 bubbles, and afterwards, the generated CO2 bubbles waves will timely explode before dissolution due to triggering by therapeutic ultrasound waves. Related bio-effects (e.g., cavitation, mechanical, shock waves, etc) caused by CO2 bubbles' explosion effectively induce instant necrosis of panc-1 cells and blood vessel destruction within panc-1 tumor, and consequently inhibit the growth of panc-1 solid tumor, simultaneously minimizing the side effects to normal organs. This new physiotherapy employing CO2 bubbling-based 'nanobomb' system promises significant potentials in targetedly suppressing tumors, especially for those highly deadly cancers.

Role of microRNAs in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is the fifth most common cancer and the third leading cause of cancer related death worldwide. HCC develops through a multistep process that involves genetic and epigenetic changes. In addition to genetic and epigenetic mechanisms, recent studies have shown that microRNAs (miRNAs) play essential roles in hepatocellular carcinogenesis through the post-transcriptional regulation of tumor associated-genes. In this review, we summarize the role of miRNAs in HCC and its microenvironment, and discuss the implications for HCC therapy.

Differential diagnosis of gallbladder wall thickening: the usefulness of contrast-enhanced ultrasound.

he purpose of this study was to evaluate the usefulness of contrast-enhanced ultrasound (CEUS) in the differential diagnosis of gallbladder wall (GBW) thickening and determine the predictors of malignant GBW thickening. One hundred fifty-nine patients with GBW thickening, including 76 men and 83 women, from eight institutions were enrolled. CEUS was performed after injection of a sulfur hexafluoride microbubble-based ultrasound contrast agent. Multiple logistic regression analysis was used to reveal independent predictor sassociated with malignant GBW thickening. The final diagnoses were 48 gallbladder carcinomas and 111 benign gallbladder diseases.Maximal thicknesses of the GBW in malignant and benign GB Wthickening were 17.3 ± 5.2 (6 ­ 30) mm and 8.6 ± 5.1 (4 ­ 26) mm respectively (p , 0.001). CEUS revealed significant differences in intralesional vessels, enhancement homogeneity, time to hypo-enhancement, inner layer discontinuity, outer layer discontinuity and adjacent liver involvement (all p-values , 0.05) between malignant and benign GBW thickening. Patient age . 46.5 y, focal GBW thickening, inner layer discontinuity and outer layer discontinuity were found to be associated with malignancy by multiple logistic regression analysis (all p-values , 0.05). Receiver operating characteristic curve analysis revealed Az values for patient age, focal GBW thickening, inner wall discontinuity and outer wall discontinuity of 0.709 (95%confidence interval [CI]: 0.627­0.790), 0.714 (95% CI: 0.630­0.798), 0.860 (95%CI: 0.791 ­ 0.928) and 0.858 (95% CI: 0.783 ­ 0.933), respectively. CEUS is useful in the differential diagnosis between malignant and benign GBW thickening. Focal GBW thickening, inner wall discontinuity and outer wall discontinuity observed on CEUS are diagnostic clues for malignant GBW thickening.

Bipolar radiofrequency ablation for liver tumors: comparison of contrast-enhanced ultrasound with contrast-enhanced MRI/CT in the posttreatment imaging evaluation.

OBJECTIVE: The aim of the study was to assess the role of contrast-enhanced ultrasound (CEUS) in treatment response evaluation after percutaneous bipolar radiofrequency ablation (BRFA) for liver tumors. METHODS: From May 2012 to May 2014, 39 patients with 73 tumors were treated by BRFA. One month after the treatment, CEUS and CEMRI/CECT were conducted to evaluate the treatment response. The results of CEUS were compared with CEMRI/CECT. RESULTS: Of the 73 tumors ablated, eight (11.0%) were found to have residual viable tumor tissue and 65 (89.0%) were successfully ablated based on CEMRI/CECT within 1-month after ablation. CEUS detected seven of the eight residual tumors and 63 of 65 completely ablated tumors. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy of CEUS were 87.5% (7/8), 96.9% (63/65), 77.8% (7/9), 98.4% (63/64) and 95.9% (70/73), respectively. The complete ablation (CR) rates for the tumors ≤ 3.0 cm, 3.1-5.0 cm, and >5.0 cm were 96.6% (58/60), 63.6% (7/11), and 0% (0/2), respectively (P<0.001). CR rates were 94.7% (36/38) for primary liver tumors and 82.9% (29/35) for metastatic liver tumors (P=0.212), and were 97.4% (38/39) for the tumors with curative treatment intention and 79.4% (27/34) for those with palliative treatment intention (P=0.037). Major complication was not encountered in this series. CONCLUSIONS: BRFA is an effective technique of percutaneous ablation for liver tumors and CEUS can be used to assess its therapeutic effect accurately.

A drug-perfluorocarbon nanoemulsion with an ultrathin silica coating for the synergistic effect of chemotherapy and ablation by high-intensity focused ultrasound.

The synergistic effect of chemotherapy and ablation using high-intensity focused ultrasound (HIFU) is realized with a newly developed drug-delivery system. The system comprises an ultrathin silica shell surrounding a poly(lactic-co-glycolic acid) nanoemulsion core containing the drug (CPT) and a perfluorocarbon (PFOB). This nanosystem presents many advantages in drug delivery, such as excellent structural stability, high drug-loading capacity, and rapid HIFU-mediated drug release.

Acoustic radiation force impulse imaging: a new tool for the diagnosis of papillary thyroid microcarcinoma.

PURPOSE: To evaluate the diagnostic performance of ARFI imaging in differentiating between benign and malignant thyroid nodules <1 cm. MATERIALS AND METHODS: 173 pathologically proven thyroid nodules (77 benign, 96 malignant) in 157 patients were included in this study. Receiver-operating characteristic curve (ROC) analyses were performed to assess the diagnostic performance of conventional ultrasound (US) and ARFI imaging in papillary thyroid microcarcinoma (PTMC). The independent risk factors for predicting PTMC were evaluated. RESULTS: The mean SWV value of benign and malignant thyroid nodules were 2.57 ± 0.79 m/s (range: 0.90-4.92 m/s) and 3.88 ± 2.24 m/s (range: 1.49-9.00 m/s) (P = 0.000). Az for VTI elastography score was higher than that for hypoechoic, absence of halo sign, and type III vascularity (P < 0.05). The optimal cut-offs for VTI elastography score and SWV were score 4 and 3.10 m/s. Gender, hypoechoic, taller than wide, VTI elastography score ≥ 4, and SWV > 3.10 m/s had been found to be independent risk factors for predicting PTMC. CONCLUSION: ARFI elastography can provide elasticity information of PTMC quantitatively (VTQ) and directly reflects the overall elastic properties (VTI). Gender, hypoechogenicity, taller than wide, VTI elastography score ≥ 4, and SWV > 3.10 m/s are independent risk factors for predicting PTMC. ARFI elastography seems to be a new tool for the diagnosis of PTMC.

Solitary schwannoma of the gallbladder: a case report and literature review.

Schwannomas occurring in the gallbladder are extremely rare. Preoperative diagnosis of gallbladder schwannomas appears to be very difficult because they are normally asymptomatic and are often found incidentally. Until now, only five cases have been reported in the literature. To our knowledge, the contrast-enhanced ultrasound (CEUS) features of gallbladder schwannomas have not been reported before in other studies. We treated a 55-year-old male patient with gallbladder schwannoma in China. He had no symptoms, and the lesion was incidentally found by conventional ultrasound (US) when performing a health examination. The patient had normal liver function; moreover, serum carcinoembryonic antigen and alpha-fetoprotein were within the normal ranges. The lesion showed no blood flow signals on color Doppler US, and the wall beneath the lesion was intact on CEUS. The lesion was believed to be a benign entity; in addition, gallbladder adenomyomatosis was suspected. A laparoscopic cholecystectomy was performed to remove the mass. Pathological examination revealed that the tumor was mainly composed of spindle-shaped cells; neither atypical cells nor signs of malignancy were found. Immunohistochemical staining showed a strong positive S-100 protein reaction. Vimentin and CD56 staining were also positive, whereas CD34 and CD117 were negative. Finally, the lesion was diagnosed as schwannoma. Herein, we report the case; the associated literature is also reviewed.

The safety and treatment response of combination therapy of radioimmunotherapy and radiofrequency ablation for solid tumor: a study in vivo.

OBJECTION: To investigate the safety and treatment response of radioimmunotherapy (RIT) in combination with radiofrequency ablation (RFA) for the treatment of VX2 tumor on rabbit. MATERIALS AND METHODS: A total of 36 rabbits bearing VX2 tumor on the thigh were randomly assigned into 3 groups (group I: 1-2 cm; group II: 2-3 cm; group III: 3-4 cm) and 4 subgroups (A: as control, just puncture the tumor using the RFA electrode without power output; B: RFA alone; C: 131I-chTNT intratumoral injection alone; D: RFA+131I-chTNT intratumoral injection 3 days later). The variation of blood assay, weight and survival among different groups and subgroups were used to assess the treatment safety. Ultrasound (US) was used to monitor and assess the tumor response after treatment. RESULTS: According to the results of the weight and the blood assay among different groups, subgroups, and at two time points (one day before and the 16th day after treatment), no damages to the liver, kidney function and myelosuppression resulting from the treatment were found. No significant differences in survivals among the four subgroups (p = 0.087) were found. In addition, 131I-chTNT did not show significant inhibition effect on VX2 tumor progression according to US measurements. CONCLUSION: 131I-chTNT intratumoral injection alone or in combination with RFA is relatively safe for rabbit without significant toxicity and shows no significant effect on the survival. The treatment response is not as satisfactory as anticipated.

Conventional US, US elasticity imaging, and acoustic radiation force impulse imaging for prediction of malignancy in thyroid nodules.

PURPOSE: To evaluate conventional ultrasonography (US), US elasticity imaging (EI), and acoustic radiation force impulse (ARFI) imaging in thyroid nodule malignancy prediction. MATERIALS AND METHODS: This prospective study was institutional review board approved; informed consent was obtained. Study included 375 patients (mean age, 51 years; range, 18-75 years) with 441 pathologically proven thyroid nodules. In 281 women (mean age, 50 years; range, 18-75 years) and 94 men (mean age, 53 years; range, 18-74 years), conventional US, EI, Virtual Touch tissue imaging (VTi; Siemens, Mountain View, Calif), and Virtual Touch tissue quantification (VTq; Siemens) of ARFI imaging were performed for each nodule. Multivariate logistic regression analysis was performed to assess 17 independent variables for malignancy prediction. Diagnostic performance was evaluated with receiver operating characteristic (ROC) curve analysis. RESULTS: There were 325 benign and 116 malignant nodules. Marked hypoechogenicity (odds ratio [OR]: 83.88; 95% confidence interval [CI]: 17.81, 394.99) was the strongest independent predictor for thyroid malignancy, followed by shape taller than wide (OR: 8.69; 95% CI: 2.87, 26.31), VTi (OR: 6.54; 95% CI: 3.61, 11.88), moderate hypoechogenicity (OR: 3.98; 95% CI: 1.13, 14.05), poorly defined margin (OR: 3.27; 95% CI: 1.22, 8.77), female sex (OR: 2.55; 95% CI: 1.33, 4.91), coarse background of surrounding thyroid tissue (OR: 2.01; 95% CI: 1.12, 3.62), and VTq (OR: 1.78; 95% CI: 1.28, 2.47) (all P < .05). EI was not significantly associated with thyroid malignancy (P = .855). Area under the ROC curve (Az) for VTq and VTi was higher than that with other significant independent variables. Az, sensitivity, and specificity were 0.91 (95% CI: 0.87, 0.94) and 0.86 (95% CI: 0.82, 0.90), 80% and 71.6%, and 93.8% and 83.4%, respectively, for VTi and VTq. VTq of at least 2.87 m/sec and VTi of at least grade IV were the best cutoff values for malignant thyroid nodules. CONCLUSION: ARFI imaging is promising for malignant thyroid nodule prediction, with higher diagnostic performance than conventional US or EI. ARFI can be used to supplement conventional US to diagnose thyroid nodules in patients referred for surgery.

A continuous tri-phase transition effect for HIFU-mediated intravenous drug delivery.

Aiming at substantially enhanced efficacy and biosafety of clinical HIFU therapy, a natural solid medium, L-menthol (LM), characteristic of mild and controllable "solid-liquid-gas" (SLG) tri-phase transition, was adopted, instead of those conventional explosive liquid-gas (LG) bi-phase transitional media, in constructing a multifunctional theranostic system. Owing to the continuous and controllable characteristics of SLG tri-phase transition, such a novel tri-phase transition-based theranostic system has been demonstrated of the repeatedly enhanced HIFU efficacy ex vivo and in vivo under once intravenous injection and the significantly improved treatment precision, controllability and biosafety when comparing to the traditional bi-phase transition medium, perfluorohexane (PFH), thus promising great application potential in clinical HIFU treatment. Moreover, this theranostic system has been demonstrated a long blood-circulation lifetime and continuous accumulation in tumor in 24 h, which is very beneficial for the enhanced tumor ablation in vivo along with SLG tri-phase transition. More importantly, after loading multiple model drugs and real drug, such a theranostic system presents a HIFU-mediated temperature-responsive drug release property, and depending on the versatile miscibility of LM, co-loadings with hydrophobic and hydrophilic drugs are also achieved, which provides the possibility of synergistic treatment combining HIFU therapy and chemotherapy.

Sorafenib blocks the HIF-1α/VEGFA pathway, inhibits tumor invasion, and induces apoptosis in hepatoma cells.

Hypoxia and hypoxia-driven angiogenesis play an important role on the recurrence of hepatocellular carcinoma after insufficient radiofrequency ablation. The hypoxia-inducible factor (HIF)-1α/vascular endothelial growth factor-A (VEGFA) pathway plays an important part in this process. Sorafenib is a multikinase inhibitor with activity against several receptor tyrosine kinases. However, it is unclear whether sorafenib can affect the HIF-1α/VEGFA pathway. Here, we explore whether sorafenib affects HIF-1α and the change of invasion ability in this process. In this experiment, the control group, cobalt chloride (CoCl2)-treated group, sorafenib-treated group, and cobalt chloride combined with sorafenib-treated group were adopted. Western blot and PCR were performed to detect the protein and mRNA expression of HIF-1α and VEGFA in different groups. Transwell assay was used to test the changes of invasion ability. Flow cytometry was adopted to detect the apoptotic role of sorafenib on hepatoma cells. Cobalt chloride upregulated the expression of HIF-1α protein, and the upregulation effect was more obvious when the concentration was increased gradually. Sorafenib inhibited cobalt-induced HIF-1α and VEGFA expression in hepatoma cells. Sorafenib decreased the tumor cell invasiveness induced by cobalt chloride in vitro. Sorafenib inhibited cell proliferation and induced apoptosis in hepatoma cells. These results showed that sorafenib was an effective inhibitor of the HIF-1α/VEGFA pathway, which can provide new insight into the mechanism of its anticancer activity.

Management of hepatocellular carcinoma: The role of contrast-enhanced ultrasound.

Hepatocellular carcinoma (HCC) is the sixth most common neoplasm and the third cause of cancer death worldwide. Contrast enhanced ultrasound (CEUS) has been applied for more than ten years and plays increasingly important roles in the management of HCC. On the basis of the Guideline and Good Clinical Practice Recommendations for CEUS in the liver-update 2012 and related literature about the management of HCC, we summarize the main roles and applications of CEUS in the management of HCC, including HCC surveillance, diagnosis, CEUS-guided treatment, treatment response evaluation and follow-up. The diagnostic algorithm for HCC is also suggested. Meanwhile, the comparisons between CEUS and contrast enhanced computed tomography/magnetic resonance imaging (CECT/CEMRI) in these areas are made. Although CEUS is subject to the same limitation as ordinary US and is inferior to CECT/CEMRI in some aspects, CEUS has proved to be of great value in the management of HCC with inherent advantages, such as sufficient high safety profile making it suitable for patients with renal failure or allergic to iodine, absence of radiation, easy reproducibility and high temporal resolution. The tremendous application of CEUS to the diagnosis and treatment of HCC provides more opportunities for patients with HCC diagnosed at different stages.

A combined "RAFT" and "Graft From" polymerization strategy for surface modification of mesoporous silica nanoparticles: towards enhanced tumor accumulation and cancer therapy efficacy.

A novel modification route integrating the copolymers of positive charged quaternary amines and polyethylene glycol (PEG) units using a combination of reversible addition-fragmentation chain-transfer polymerization (RAFT) and "Graft From" strategy, has been proposed and developed, for the first time, to decorate the surface of mesoporous silica nanoparticles (MSNs). These MSNs are shown to have a greatly reduced hydrodynamic particle size in physiological solution. It is demonstrated that such an efficient copolymer surface modification strategy, resulting in PEG coating with high positive zeta potential, can achieve a nearly 2-fold enhanced permeability and retention (EPR) effect, and longer blood half-life compared to coating with PEG only. Besides, the in vivo results demonstrated that this surface modification strategy could lead to a higher efficacy of doxorubicin (DOX) drug delivery and greater suppression of side effects compared to the free drug. Based on this novel strategy of combining "RAFT" and "Graft From" polymerization, it is anticipated that this efficient modification of tumor-specific targeting of MSNs can be widely used in future nanomedicine research.

An intelligent nanotheranostic agent for targeting, redox-responsive ultrasound imaging, and imaging-guided high-intensity focused ultrasound synergistic therapy.

A novel multifunctional nanotheranostic agent with targeting, redox-responsive ultrasound imaging and ultrasound imaging-guided high-intensity focused ultrasound (HIFU) therapy (MSNC-PEG-HA(SS)-PFH, abbreviated as MPH(SS)-PFH) capabilities is developed. The redox-responsive guest molecule release and ultrasound imaging functions can be both integrated in such a "smart" theranostic agent, which is accomplished by the redox-triggered transition from the crosslinking state to retrocrosslinking state of the grafted polyethylene glycol-disulfide hyaluronic acid molecules on the particle surface when reaching a reducing environment in vitro. More importantly, under the tailored ultrasound imaging guiding, in vivo Hela tumor-bearing nude mice can be thoroughly and spatial-accurately ablated during HIFU therapy, due to the targeted accumulation, responsive ultrasound imaging guidance and the synergistic ablation functions of nanotheranostic agent MPH(SS)-PFH in the tumors. This novel multifunctional nano-platform can serve as a promising candidate for further studies on oncology therapy, due to its high stability, responsive and indicative ultrasound imaging of tumors, and enhanced HIFU therapeutic efficiency and spatial accuracy under ultrasound-guidance.

Liver specific phase of perflubutane microbubbles contrast-enhanced ultrasound for potential use in the diagnosis of focal liver lesions.

The purpose of this study was to distinguish passive retention of microbubbles in liver sinusoids from active intracellular retaining, and to provide further evidence of post vascular liver specific phase of perflubutane microbubble contrast-enhanced ultrasound for potential use in the detection of focal liver lesions. Fifty-four rats were divided into three groups: saline group (G1, as a control, n=6), sulphur hexafluoride microbubble group (G2, n=24) and perflubutane microbubble group (G3, n=24). The livers were perfused at 2, 5, 10 and 20 min after injection of perflubutane or sulphur hexafluoride gas-filled microbubbles or normal saline. Changes in contrast-enhanced images within the liver were quantified. After perfusion, the echogenicity of the liver in the saline group increased from -49.44 ± 0.08 dB to - 44.37 ± 1.02 dB (p< 0.05). In G2, the enhancement decreased significantly after perfusion at 2 and 5 min, and increased at 10 and 20 min (all p< 0.05). In G3, the enhancement decreased from -18.05 ± 2.28 dB to - 26.76 ± 1.69 dB only at 2 min after perfusion (p < 0.05). Perflubutane microbubbles provided a post vascular liver specific phase on contrast-enhanced ultrasound, which could begin as early as 5 min after administration of the contrast agent in rats. This study suggests that perflubutane microbubbles could result in a liver specific phase and provide valuable information for diagnosis and detection of focal liver lesions which may improve the efficiency of contrast-enhanced ultrasound in clinical diagnoses of liver diseases.