The effect and safety assessment of monitoring ethanol concentration in exhaled breath combined with intelligent control of renal pelvic pressure on the absorption of perfusion fluid during flexible ureteroscopic lithotripsy.

PURPOSE: Previously, we designed a ureteral access sheath with the capability of renal pelvic pressure (RPP) measurement and a medical perfusion and aspiration platform, allowing for the intelligent control of RPP. However, the effect of different RPP levels on perfusion fluid absorption remains unclear. This randomized controlled trial aimed to investigate the effects of exhaled ethanol concentration monitoring and intelligent pressure control on perfusion fluid absorption during flexible ureteroscopic lithotripsy. METHODS: Eighty patients scheduled for flexible ureteroscopic lithotripsy were randomly divided into four groups. In groups A, B, and C, the RPPs were set at 0, - 5, and - 10 mmHg, respectively. Group D was regarded as the controls with unfixed RPP. Isotonic saline containing 1% ethanol was used as the irrigation fluid, with an average irrigation flow rate of 100 mL/min. The primary outcome of this study was the absorption of perfusion fluid that was calculated based on the exhaled ethanol concentration. The secondary outcomes included duration of operation and amounts of perfusion fluid used. Postoperative complications, pre- and postoperative renal function, infection markers, and blood gas analysis were also recorded for safety assessment. RESULTS: In all, 76 patients were involved in this study, whose demographic characteristics and preoperative conditions were comparable among groups. Under the same perfusion flow rate, the groups with fixed RPP exhibited reduced absorption of perfusion fluid, duration of operation, and perfusion volume. In particular, the lowest values were observed in group C (RPP = - 10 mmHg). In contrast to the unfixed RPP group, no considerable difference were observed in levels of BUN, Scr, WBC, CRP, and blood gas values among the fixed RPP groups. Moreover, postoperative complications showed no significant difference among groups. CONCLUSION: In flexible ureteroscopic lithotripsy, the groups with fixed RPP had less absorption of perfusion fluid and perfusion volume, shorter duration of surgery, and higher safety than the unfixed group.

Comparative diagnostic accuracy of the IOTA SRR and LR2 scoring systems for discriminating between malignant and Benign Adnexal masses by junior physicians in Chinese patients: a retrospective observational study.

BACKGROUND: The accuracy of ultrasound in distinguishing benign from malignant adnexal masses is highly correlated with the experience of ultrasound physicians. In China, most of ultrasound differentiation is done by junior physicians. PURPOSE: To compare the diagnostic performance of the International Ovarian Tumour Analysis (IOTA) Simple Rules Risk (SRR) and IOTA Logistic Regression Model 2 (LR2) scoring systems in Chinese patients with adnexal masses. METHODS: Retrospective analysis of ovarian cancer tumor patients who underwent surgery at a hospital in China from January 2016 to December 2021. Screening patients with at least one adnexal mass on inclusion and exclusion criteria. Two trained junior physicians evaluated each mass using the two scoring systems. A receiver operating characteristic curve was used to test the diagnostic performance of each system. RESULTS: A total of 144 adnexal masses were retrospectively collected. Forty masses were histologically diagnosed as malignant. Compared with premenopausal women, postmenopausal women had a much higher rate of malignant masses. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) of the SRR was 97.5% (95% CI: 86.8 -99.9%), 82.7% (95% CI: 74.0 -89.4%), 68.4% (95% CI: 58.7 -76.8%) and 98.9% (95% CI: 92.5 -99.8%). The sensitivity, specificity, PPV, NPV of the LR2 were 90.0% (95% CI: 76.5 -97.2%), 89.4% (95% CI: 81.9 -94.6%), 76.6% (95% CI: 65.0 -85.2%), and 95.9% (95% CI: 90.2 -98.3%). There was good agreement between two scoring systems, with 84.03% total agreement and a kappa value of 0.783 (95% CI: 0.70-0.864). The areas under the curve for predicting malignant tumours using SRR and LR2 were similar for all patients (P > 0.05 ). CONCLUSION: The two scoring systems can effectively distinguish benign from malignant adnexal masses. Both scoring systems have high diagnostic efficacy, and diagnostic efficacy is stable, which can provide an important reference for clinical decision making.

Transcriptome analysis revealed detoxification gene expression changes in Tetranychus cinnabarinus challenged with ethyl oleate.

Natural acaricides are potential biorational mite control alternatives to conventional chemical acaricides. However, little is known about the molecular mechanism of defense response to natural acaricides in mites. We previously reported significant acaricidal properties of ethyl oleate (EO) against Tetranychus cinnabarinus (here referred to as a sibling species of two-spotted spider mite, Tetranychus urticae), a highly polyphagous pest devastating crops in fields and greenhouses worldwide. In this study, we explored the molecular responses of T. cinnabarinus exposed to EO using RNA-Seq and differentially expressed gene (DEG) analysis. A total of 131, 185, and 154 DEGs were identified in T. cinnabarinus after 1, 6, and 24 h of EO treatment. In addition, 36 putative detoxification-related DEGs, including 10 cytochrome P450s (P450s), three glutathione S-transferases (GSTs), nine UDP-glycosyltransferases (UGTs), eight esterases (ESTs), and six ATP-binding cassette transporters (ABC transporters), were identified. Interestingly, the upregulation of these detoxification-related genes might be the main defense response of T. cinnabarinus exposed to EO. A quantitative real-time PCR analysis indicated that the expression profiles of 19 random DEGs were consistent with the RNA-Seq results. These findings serve as valuable information for a better understanding of the acaricide-mite interaction and molecular mechanisms involved in the defense response of T. cinnabarinus against EO.

Identification and expression profile of odorant-binding proteins in the parasitic wasp Microplitis pallidipes using PacBio long-read sequencing.

Microplitis pallidipes Szépligeti (Hymenoptera: Braconidae) is an important parasitic wasp of second and third-instar noctuid larvae such as the insect pests Spodoptera exigua, Spodoptera litura, and Spodoptera frugiperda. As in other insects, M. pallidipes has a chemosensory recognition system that is critical to foraging, mating, oviposition, and other behaviors. Odorant-binding proteins (OBPs) are important to the system, but those of M. pallidipes have not been determined. This study used PacBio long-read sequencing to identify 170,980 M. pallidipes unigenes and predicted 129,381 proteins. Following retrieval of possible OBP sequences, we removed those that were redundant or non-full-length and eventually cloned five OBP sequences: MpOBP2, MpOBP3, MpOBP8, MpOBP10, and MpPBP 429, 429, 459, 420, and 429 bp in size, respectively. Each M. pallidipes OBP had six conserved cysteine residues. Phylogenetic analysis revealed that the five OBPs were located at different branches of the phylogenetic tree. Additionally, tissue expression profiles indicated that MpOBP2 and MpPBP were mainly expressed in the antennae of male wasps, while MpOBP3, MpOBP8, and MpOBP10 were mainly expressed in the antennae of female wasps. MpOBP3 was also highly expressed in the legs of female wasps. Temporal profiles revealed that the expression of each M. pallidipes OBP peaked at different days after emergence to adulthood. In conclusion, we identified five novel odorant-binding proteins of M. pallidipes and demonstrated biologically relevant differences in expression patterns.

Title: Identification et profil d'expression des protéines de liaison aux odeurs chez la guêpe parasite Microplitis pallidipes à l'aide du séquençage à lecture longue PacBio. Abstract: Microplitis pallidipes Szépligeti (Hymenoptera : Braconidae) est une importante guêpe parasite des larves de noctuelles de deuxième et troisième stades telles que les insectes ravageurs Spodoptera exigua, Spodoptera litura et Spodoptera frugiperda. Comme d'autres insectes, M. pallidipes possède un système de reconnaissance chimiosensoriel, essentiel à la recherche de nourriture, à l'accouplement, à la ponte et à d'autres comportements. Les protéines de liaison aux odeurs (PLO) sont importantes pour le système, mais celles de M. pallidipes n'ont pas été déterminées. Cette étude a utilisé le séquençage à lecture longue PacBio pour identifier 170 980 unigènes de M. pallidipes et prédit 129 381 protéines. Après la récupération des séquences de PLO possibles, nous avons supprimé celles qui étaient redondantes ou pas de pleine longueur et avons finalement cloné cinq séquences de PLO, MpOBP2, MpOBP3, MpOBP8, MpOBP10 et MpPBP, respectivement de taille 429, 429, 459, 420 et 429 pb. Chaque PLO de M. pallidipes avait six résidus de cystéine conservés. L'analyse phylogénétique a révélé que les cinq PLO étaient situés à différentes branches de l'arbre phylogénétique. De plus, les profils d'expression tissulaire ont indiqué que MpOBP2 et MpPBP étaient principalement exprimés dans les antennes des guêpes mâles, tandis que MpOBP3, MpOBP8 et MpOBP10 étaient principalement exprimés dans les antennes des guêpes femelles. MpOBP3 était également fortement exprimé dans les pattes des guêpes femelles. Les profils temporels ont révélé que l'expression de chaque PLO de M. pallidipes culminait à différents jours après l'émergence à l'âge adulte. En conclusion, nous avons identifié cinq nouvelles protéines de liaison aux odeurs de M. pallidipes et démontré des différences biologiquement pertinentes dans les profils d'expression.

Decline of three farmland pest species in rapidly urbanizing landscapes.

Urbanization is a pressing challenge for earth's humans because it is changing not only natural environments but also agricultural lands. Yet, the consequences of cropland loss on pest insect populations that largely depend on these habitats remain largely unclear. We used a 17-year data set to investigate the dynamics of three moth pest species (i.e., striped stem borer, yellow stem borer, and pink stem borer) and their driving forces across the largest mega-urban region of China. Total abundance of three pest species is declined by about 80%, which was strongly associated with cropland loss during rapid urbanization. Our findings indicate that not only the increasing conversion of natural areas to human-dominated landscapes but also that of agricultural lands to urban landscapes can be critical to insect populations. It is therefore essential to monitor and understand the insect dynamics in rapidly urbanizing regions, which are currently found in many developing countries worldwide.

Effect of MT2A on apoptosis and proliferation in HL60 cells.

Although accumulating evidence has revealed that metallothioneins (MTs) and its family member MT2A are strongly linked to the risk of various solid tumors, researches on the occurrence and development of acute myeloid leukemia (AML) have rarely been investigated. Here, we constructed a lentiviral vector with MT2A over-expression and the interfering plasmids with MT2A expression inhibition to study the influence of MT2A on the bioactivities of HL60 cells. After cells were infected with a lentiviral vector containing the MT2A gene, both transcription and translation levels of MT2A were significantly increased in the over-expressed group in comparison with control groups. In vitro experiments, all results demonstrated that cell reproductive capacity was inhibited, but cell apoptosis rate was significantly increased. Together, the expression of apoptosis-related protein Bcl2 was remarkably reduced, while a high expression level of Bax protein was detected. Further experiments revealed that up-regulation of MT2A induced cell apoptosis and promoted G2/M phase arrest. The mechanism may be associated with down-regulated p-IκB-α and cyclinD1 expression and up-regulated IκB-α expression in the nuclear factor-kappaB (NF-κB) pathway. On the contrary, MT2A expression was down-regulated by interfering plasmids. We found that cell proliferative potential was notably increased in the interfering group compared with the negative and untreated group. What's more, MT2A may be closely related to AML cell proliferation and function via the NF-κB signal pathway.

Plants impact cellular immunity of caterpillars to an entomovirus.

BACKGROUND: Tri-trophic interactions among plants, insect herbivores and entomopathogens are one of the hot topics in ecology. Although plants have been shown to impact the interactions between herbivores and entomopathogens, it is still unclear how plants affect the cellular immunity of herbivores to entomopathogens. RESULTS: The number of hemocytes and the proportion of two main cell types (granular hemocytes and plasmatocytes), plasmatocyte-spreading rate, apoptosis rate, two Spodoptera exigua caspase (SeCasp-1, SeCasp-5) activities and gene expressions were all higher and the activities and gene expression of S. exigua inhibitor of apoptosis protein (SeIAP) were lower in nucleopolyhedrovirus (NPV)-infected caterpillars fed Ipomoea aquatica than those fed other plants or artificial diet. Scanning electron microscopy images were consistent with molecular patterns of immune responses. CONCLUSION: This study suggests that host plants affect the immune responses of herbivores to entomopathogens by manipulating the composition, morphology and apoptosis of herbivore hemocytes, which sheds light on the mechanisms that allow host plants to influence multi-trophic interactions. © 2021 Society of Chemical Industry.

Highly specific nanobody against herbicide 2,4-dichlorophenoxyacetic acid for monitoring of its contamination in environmental water.

2,4-dichlorophenoxyacetic acid (2,4-D), a widely used herbicide, is a small organic chemical pollutant in the environment. To develop a nanobody-based immunoassay for monitoring trace levels of 2,4-D, a step-wise strategy for the generation of nanobodies highly specific against this small chemical was employed. Firstly, we synthesized three novel haptens mimicking 2,4-D and assessed their influence on the sensitivity and specificity of the existing antibody-based assay. Polyclonal antibodies (pAb) from rabbits showed good sensitivity and moderate specificity for 2,4-D, pAb from llama based on selected haptens showed similar performance when compared to those from rabbits. Secondly, nanobodies derived from llama were generated for 2,4-D by an effective procedure, including serum monitoring and one-step library construction. One nanobody, NB3-9, exhibited good sensitivity against 2,4-D (IC50 = 29.2 ng/mL) had better specificity than the rabbit pAb#1518, with no cross-reactivities against the 2,4-D analogs tested. Thirdly, one-step fluorescent enzyme immunoassay (FLEIA) for 2,4-D based on a nanobody-alkaline phosphatase (AP) fusion was developed with IC50 of 1.9 ng/mL and a linear range of 0.4-8.6 ng/mL. Environmental water samples were analyzed by FLEIA and LC-MS/MS for comparison, and the results were consistent between both methods. Therefore, the proposed step-wise strategy from hapten design to nanobody-AP fusion production was successfully conducted, and the resulting nanobody based FLEIA was demonstrated as a convenient tool to monitor 2,4-D residuals in the environment.

Development, Screening, and Validation of Camelid-Derived Nanobodies for Neuroscience Research.

Nanobodies (nAbs) are recombinant antigen-binding variable domain fragments obtained from heavy-chain-only immunoglobulins. Among mammals, these are unique to camelids (camels, llamas, alpacas, etc.). Nanobodies are of great use in biomedical research due to their efficient folding and stability under a variety of conditions, as well as their small size. The latter characteristic is particularly important for nAbs used as immunolabeling reagents, since this can improve penetration of cell and tissue samples compared to conventional antibodies, and also reduce the gap distance between signal and target, thereby improving imaging resolution. In addition, their recombinant nature allows for unambiguous definition and permanent archiving in the form of DNA sequence, enhanced distribution in the form of sequences or plasmids, and easy and inexpensive production using well-established bacterial expression systems, such as the IPTG induction method described here. This article will review the basic workflow and process for developing, screening, and validating novel nAbs against neuronal target proteins. The protocols described make use of the most common nAb development method, wherein an immune repertoire from an immunized llama is screened via phage display technology. Selected nAbs can then be taken through validation assays for use as immunolabels or as intrabodies in neurons. © 2020 Wiley Periodicals LLC. Basic Protocol 1: Total RNA isolation from camelid leukocytes Basic Protocol 2: First-strand cDNA synthesis; VH H and VH repertoire PCR Basic Protocol 3: Preparation of the phage display library Basic Protocol 4: Panning of the phage display library Basic Protocol 5: Small-scale nAb expression Basic Protocol 6: Sequence analysis of selected nAb clones Basic Protocol 7: Nanobody validation as immunolabels Basic Protocol 8: Generation of nAb-pEGFP mammalian expression constructs Basic Protocol 9: Nanobody validation as intrabodies Support Protocol 1: ELISA for llama serum testing, phage titer, and screening of selected clones Support Protocol 2: Amplification of helper phage stock Support Protocol 3: nAb expression in amber suppressor E. coli bacterial strains.

Imaging and Contrast-enhanced Ultrasound features of duodenal neuroendocrine tumor: A case report.

Duodenal neuroendocrine tumors are rare neoplasms arising from endocrine cells. Here we present a case of 32-year-old woman with Duodenal neuroendocrine tumors, report the imaging and contrast-enhanced Ultrasound (CEUS) features and review previous literatures of neuroendocrine tumors, which may be valuable for the differential diagnosis of duodenal neoplasms.

Global synthesis of effects of plant species diversity on trophic groups and interactions.

Numerous studies have demonstrated that plant species diversity enhances ecosystem functioning in terrestrial ecosystems, including diversity effects on insects (herbivores, predators and parasitoids) and plants. However, the effects of increased plant diversity across trophic levels in different ecosystems and biomes have not yet been explored on a global scale. Through a global meta-analysis of 2,914 observations from 351 studies, we found that increased plant species richness reduced herbivore abundance and damage but increased predator and parasitoid abundance, predation, parasitism and overall plant performance. Moreover, increased predator/parasitoid performance was correlated with reduced herbivore abundance and enhanced plant performance. We conclude that increasing plant species diversity promotes beneficial trophic interactions between insects and plants, ultimately contributing to increased ecosystem services.

Immunity of an insect herbivore to an entomovirus is affected by different host plants.

BACKGROUND: Interactions between herbivorous insects and entomoviruses may depend on host plant, perhaps mediated through changes in herbivore innate immunity. RESULTS: Caterpillars (Spodoptera exigua) fed Glycine max had high viral loads and low melanization rates together with low melanization enzyme [PO, DDC, TH] activities and gene expressions. Caterpillars fed Ipomoea aquatica had low viral loads and high melanization, gene activities and gene expressions while those fed Brassica oleracea or artificial diet had intermediate levels of each. Melanization rates were negatively correlated with viral loads and positively correlated with activity and expression of each of the three enzymes. Some diet effects on enzymes were constitutive because the same diets led to low (G. max) or high (I. aquatica) melanization related gene activities and expressions without infection. CONCLUSION: Diet influences the interactions between insect herbivores and viruses by shaping the innate immune response both at the onset of infection and afterwards as viral loads accumulate over a period of days. In addition, diets that lead to low viral loads are associated with high activities and gene expressions of a variety of melanization related enzymes suggesting a common causative mechanism. © 2019 Society of Chemical Industry.

Comparison of Clinical Characteristics and Outcomes of Pediatric and Adult Patients with Coronavirus Disease 2019 in Shenzhen, China / 生物医学与环境科学（英文）

Objective@#Here we aimed to investigate the difference in clinical characteristics and outcomes between pediatric and adult patients with COVID-19.@*Methods@#A total of 333 consecutive patients with laboratory-confirmed SARS-CoV-2 infection treated in the departments of Internal medicine of Shenzhen Third People's Hospital from January 11 @*Results@#Compared with adult patients, pediatric patients had a shorter time of symptom onset to hospitalization than adults [median time, 1 ( @*Conclusion@#Pediatric patients with COVID-19 had milder or less clinical symptoms, less evident pulmonary imaging changes, better prognosis, and shorter length of hospital stay.

Development of a Simple Pretreatment Immunoassay Based on an Organic Solvent-Tolerant Nanobody for the Detection of Carbofuran in Vegetable and Fruit Samples.

Nanobodies are one-tenth the size of conventional antibodies and are naturally obtained from the atypical heavy-chain-only antibodies present in camelids. Their small size, high solubility, high stability, and strong resilience to organic solvents facilitate their use as novel analytical reagents in immunochemistry. In this study, specific nanobodies against pesticide carbofuran were isolated and characterized from an immunized library via phage display platform. We further established an indirect competitive enzyme-linked immunosorbent assay (ELISA) using nanobody Nb316 to detect carbofuran in vegetable and fruit samples. The results showed a half-maximal inhibitory concentration (IC50) of 7.27 ng/mL and a detection limit of 0.65 ng/mL. A simplified sample pretreatment procedure omitting the evaporation of organic solvent was used. The averaged recovery rate of spiked samples ranged between 82.3% and 103.9%, which correlated with that of standard UPLC-MS/MS method. In conclusion, a nanobody with high specificity for carbofuran was characterized, and a nanobody-based sensitive immunoassay for simple and rapid detection of carbofuran in real samples was validated.

Modulating Linker Composition of Haptens Resulted in Improved Immunoassay for Histamine.

Histamine (HA) is an important food contaminant generated during food fermentation or spoilage. However, an immunoassay for direct (derivatization free) determination of HA has rarely been reported due to its small size to induce the desired antibodies by its current hapten-protein conjugates. In this work, despite violating the classical hapten design criteria which recommend introducing a linear aliphatic (phenyl free) linker into the immunizing hapten, a novel haptens, HA-245 designed and synthesized with a phenyl-contained linker, exhibited significantly enhanced immunological properties. Thus, a quality-improved monoclonal antibody (Mab) against HA was elicited by its hapten-carrier conjugates. Then, as the linear aliphatic linker contained haptens, Hapten B was used as linker-heterologous coating haptens to eliminate the recognition of linker antibodies. Indirect competitive ELISA (ic-ELISA) was developed with a 50% inhibition concentration (IC50) of 0.21 mg/L and a limit of detection (LOD) of 0.06 mg/L in buffer solution. The average recoveries of HA from spiked food samples for this ic-ELISA ranged from 84.1% and 108.5%, and the analysis results agreed well with those of referenced LC-MS/MS. This investigation not only realized derivatization-free immunoassay for HA, but also provided a valuable guidance for hapten design and development of immunoassay for small molecules.

A toolbox of nanobodies developed and validated for use as intrabodies and nanoscale immunolabels in mammalian brain neurons.

Nanobodies (nAbs) are small, minimal antibodies that have distinct attributes that make them uniquely suited for certain biomedical research, diagnostic and therapeutic applications. Prominent uses include as intracellular antibodies or intrabodies to bind and deliver cargo to specific proteins and/or subcellular sites within cells, and as nanoscale immunolabels for enhanced tissue penetration and improved spatial imaging resolution. Here, we report the generation and validation of nAbs against a set of proteins prominently expressed at specific subcellular sites in mammalian brain neurons. We describe a novel hierarchical validation pipeline to systematically evaluate nAbs isolated by phage display for effective and specific use as intrabodies and immunolabels in mammalian cells including brain neurons. These nAbs form part of a robust toolbox for targeting proteins with distinct and highly spatially-restricted subcellular localization in mammalian brain neurons, allowing for visualization and/or modulation of structure and function at those sites.

Isolation of Bactrian Camel Single Domain Antibody for Parathion and Development of One-Step dc-FEIA Method Using VHH-Alkaline Phosphatase Fusion Protein.

A heavy chain variable fragment of heavy chain only antibodies derived from camelids termed VHH shows beneficial characteristics for immunoassay in terms of high sensitivity, outstanding stability and ease in expression. In the present study, we isolated six VHHs from phage display library against parathion, which is a widely used organophosphorus pesticide with high toxicity and persistence. One of six selected VHHs named VHH9, showed highest specificity and superior thermo-stability. A VHH9-alkaline phosphatase (AP) fusion was constructed and used to establish a one-step direct competitive fluorescence enzyme immunoassay (dc-FEIA) with a half maximal inhibitory concentration (IC50) of 1.6 ng/mL and a limit of detection of 0.2 ng/mL which was 4-fold or 3-fold higher sensitivity than direct competitive enzyme-linked immunoassay (dc-ELISA) and indirect competitive enzyme-linked immunoassay (ic-ELISA) for parathion. Furthermore, our assay indicated a 50% reduction on operation time compared with the ic-ELISA method. The presented immunoassay was validated with spiked Chinese cabbage, cucumber, and lettuce samples, and confirmed by UPLC-MS/MS. The results indicated that the VHH-AP-based dc-FEIA is a reproducible detection assay for parathion residues in vegetable samples.

Increasing plant diversity with border crops reduces insecticide use and increases crop yield in urban agriculture.

Urban agriculture is making an increasing contribution to food security in large cities around the world. The potential contribution of biodiversity to ecological intensification in urban agricultural systems has not been investigated. We present monitoring data collected from rice fields in 34 community farms in mega-urban Shanghai, China, from 2001 to 2015, and show that the presence of a border crop of soybeans and neighboring crops (maize, eggplant and Chinese cabbage), both without weed control, increased invertebrate predator abundance, decreased the abundance of pests and dependence on insecticides, and increased grain yield and economic profits. Two 2 year randomized experiments with the low and high diversity practices in the same locations confirmed these results. Our study shows that diversifying farming practices can make an important contribution to ecological intensification and the sustainable use of associated ecosystem services in an urban ecosystem.

Dual-Targeted Microbubbles Specific to Integrin αVß3 and Vascular Endothelial Growth Factor Receptor 2 for Ultrasonography Evaluation of Tumor Angiogenesis.

Aggressive tumors are characterized by angiogenesis that promotes the migration and dissemination of tumor cells. Our aim was to develop a dual-targeted microbubble system for non-invasive evaluation of tumor angiogenesis in ultrasound. Avidinylated microbubbles were conjugated with biotinylated arginylglycylaspartic acid and vascular endothelial growth factor receptor 2 (VEGFR2) antibodies. Subcutaneous MHCC-97H liver carcinoma models were established. Non-targeted, αvß3-targeted, VEGFR2-targeted and dual-targeted microbubbles was intravenously injected in series while acquiring ultrasound images of the tumor. The microbubbles were destroyed by a high-mechanical-index pulse 4 min after the injection. Peak intensity (PI) before and after the destructive pulse was recorded to compare contrast enhancement by different microbubbles. The targeting rates of the integrin-targeted, VEGFR2-targeted and dual-targeted groups were 95.02%, 96.04% and 94.23%, respectively, with no significant differences. Tumors in all groups were significantly enhanced. The time-intensity curve indicated no significant differences in arrival time, PI, area under the curve, amplitude and mean transit time. The difference in ultrasound signal intensity before and after the destructive pulse (⊿PI) for all targeted microbubble groups was significantly greater than that for the non-targeted microbubble group (all p values < 0.05), and the difference for the dual-targeted microbubble group was significantly greater than those of both mono-targeted groups (p <0.05).