Superiorly Stable Three-Layer Air Microbubbles Generated by Versatile Ethanol-Water Exchange for Contrast-Enhanced Ultrasound Theranostics.

Microbubbles have been widely used as ultrasound contrast agents in clinical diagnosis. Moreover, most current preparation methods for microbubbles are uncontrollable, and the as-obtained microbubbles are unstable in aqueous solution or under ultrasound. Here, we report a strategy to prepare superiorly stable microbubbles with three-layer structures by the ethanol-water exchange. This versatile method can also be applied to prepare different kinds of protein microbubbles with various sizes for advanced biomedical applications. To demonstrate this, the protein air microbubbles are created, which is stable in water for several days with intact structures and exhibits excellent contrast-enhanced ultrasound imaging. Moreover, the protein air microbubbles can also deliver a mass of drugs while maintaining their stable structures, making them a platform for ultrasound imaging-guided drug delivery. The versatile protein air microbubbles have great potential for the design and application of theranostic platforms.

Choriocarcinoma Masquerading as Lung Abscess or Lung Cancer: A Case with Atypical Imaging Findings.

BACKGROUND: Choriocarcinoma is a highly malignant trophoblastic tumor. However, the awareness surrounding its atypical clinical presentation is insufficient. The presence of a solitary lung lesion without uterine lesions often leads to misdiagnosis or missed diagnosis, which in turn causes delayed treatment or even multiple metastases throughout the body. CASE PRESENTATION: We present the case of a 36-year-old female patient who was misdiagnosed with a lung abscess and received suboptimal anti-infective treatment. She then underwent left upper lobectomy and was misdiagnosed with lung cancer by abscess incision and drainage in thoracic surgery, however, the results after pleural effusion removal were suboptimal. During this time a breast nodule was found, and a large segment of the right breast was excised and misdiagnosed as breast cancer but was finally diagnosed as choriocarcinoma with multiple metastases of lung and breast. Multiple metastases were also detected in the head, liver, kidney, and bones. The patient underwent multiple adjuvant chemotherapies. The blood ß-hCG level gradually declined to normal. When we reported this case, that is, seven months after the diagnosis, the patient was still alive, and the disease was stable without progress. CONCLUSION: Choriocarcinoma with a solitary lung lesion as the first presentation and no lesions in the uterus is clinically rare. This may lead to a delay in diagnosis due to poor awareness of the disease and the appearance of multiple metastases throughout the body. Clinicians should be more aware of choriocarcinoma with an atypical presentation to reduce misdiagnosis and missed diagnosis.

Ultrasound-visualized, site-specific vascular embolization using magnetic protein microcapsules.

Imaging-guided vascular embolization is frequently performed on patients with advanced hepatocellular carcinoma (HCC) to alleviate symptoms and extend their survival time. Current operation procedures are not only painful for patients, but are also inaccurate in tumor targeting after the release of embolic agents from the catheter, leading to injury to healthy tissues simultaneously. In this study, we developed an ultrasound-visualized, site-specific vascular embolization strategy with magnetic protein microcapsules (MPMs). MPMs were fabricated using a rapid emulsification method, giving it a smooth surface and a core-shell structure. Their diameters could be controlled within 10 µm, allowing them to pass through capillaries. The core-shell structure and loading of magnetic Fe3O4 endowed MPMs with good contrast under ultrasound imaging and magnetically inducible targeting properties, as well as aggregation response even under flowing conditions. In vitro cytotoxicity and hemolysis evaluation demonstrated good biocompatibility of the MPMs. Furthermore, mock embolization showed that cell death could be induced by aggregation of the MPMs. Such a combination of real-time monitoring using ultrasound and control on targeted vascular embolization might be a breakthrough in the treatment of advanced HCC.

Multimodality imaging-guided local injection of eccentric magnetic microcapsules with electromagnetically controlled drug release.

BACKGROUND: In the past decade, trackable smart drug delivery systems have played important roles in the treatment of many diseases such as cancer because the drug carriers can be visualized through their distinct physical properties. However, it is still difficult to achieve precise drug delivery because such systems usually rely on a single imaging system. AIM: This study aimed to present a novel type of multimodality imaging-guided strategy to visualize the drug carriers of eccentric magnetic microcapsule (EMM) designed for potential treatment of hepatocellular carcinoma (HCC). METHOD AND RESULTS: The EMMs were prepared by using a three-phase microfluidic device. The as-prepared EMMs embedded with Fe3O4 nanoparticles are magnetic, with high density and acoustic impedance, allowing for visualization by magnetic resonance imaging (MRI), computed tomography (CT), and ultrasound (US) imaging during local injection. The release of drug from these EMMs can be further controlled by an external electromagnetic field (EMF). As a proof of concept, we demonstrated the process of multimodality imaging to guide local injection and the controlled release of doxorubicin (DOX) from the EMMs in a phantom. We showed that the release rate of DOX was directly correlated to the strength of the EMF. In addition, we cocultured green fluorescent protein (GFP)-transfected HeLa cancer cells with the DOX-loaded EMMs and documented their apoptosis by DOX following the release triggered by EMF. CONCLUSION: The results suggest that these EMMs serve both as contrast agents that can be visualized by multimodality imaging techniques and as smart drug delivery systems, with great potential for precision medicine.

The expression and role of MEKK3 in renal clear cell carcinoma.

To explore the relationship between Mitogen-activated protein kinase/extracellular signal-regulated kinase kinase kinase 3(MEKK3) and cell apoptosis, clinicopathology, and prognosis, we characterize the expression of MEKK3, survivin and stat3 in renal clear cell carcinoma (RCCC). The expressions were measured by RT-PCR and Western blot from 15 cases of RCCC and 15 cases of control group (CG). Protein expression was detected by tissue microarray and immunochemistry in 98 cases of RCCC, 28 cases of CG. Expression patterns were analyzed for their association with pathological factors, correlation and prognosis in RCCC. Expression of MEKK3, survivin and stat3 mRNA was significantly higher in RCCC than in CG (P < 0.01). MEKK3, survivin and stat3 expression differed significantly between pathological grade (P < 0.05) and clinical stage (P < 0.05). MEKK3 expression was positively correlated with survivin and stat3 (P < 0.01). Furthermore, we investigated the role of MEKK3 in RCCC using the technique of RNA silencing via small interfering (siRNA) in ACHN cells. The results indicated that the targeted depletion of MEKK3 caused a dramatic massive apoptotic cell death. Kaplan-Meier survival analysis showed that MEKK3 and survivin expression, pathological grade, and clinical stage reduced cumulative survival. Cox multivariate regression analysis showed that MEKK3, survivin, and clinical staging were independent prognostic factors in renal cancer (P < 0.05). MEKK3 can be used as an important marker of prognostic evaluation in RCCC. The mechanism may be closely related to cell apoptosis. Targeted therapy of MEKK3 may provide a new strategy for treatment of chemotherapeutic-resistant tumors.

Expression and prognostic role of MEKK3 and pERK in patients with renal clear cell carcinoma.

Mitogen-activated protein kinase/extracellular signal-regulated kinase kinase kinase 3 (MEKK3) is an important serine/threonine protein kinase and a member of the MAPK family. MEKK3 can effectively activate the MEK/ERK signaling pathway and promote an autocrine growth loop critical for tumor genesis, cell proliferation, terminal differentiation, apoptosis and survival. To explore the relationship between MEKK3 and cell apoptosis, clinicopathology and prognosis, we characterize the expression of MEKK3, pERK and FoxP3 in the renal clear cell carcinoma (RCCC). Protein expression was detected by tissue microarray and immunochemistry in 46 cases of RCCC and 28 control cases. Expression levels of CD3+ ,CD3+CD4+,CD3+CD8+,CD4+CD25+, CD4+CD25+ FoxP3+ were assessed by flow cytometry and analyzed for their association with pathological factors, correlation and prognosis in RCCC. Expression of MEKK3, pERK and FoxP3 was significantly up-regulated in RCCC as compared to control levels (p<0.01), associated with pathological grade (p<0.05)and clinical stage (p<0.05). CD4+CD25+ Foxp3+ Treg cells were also significantly increased in RCCC patients (p<0.05). Cox multivariate regression analysis showed that MEKK3, pERK expression and patholigical stage were independent prognostic factors in patients with RCCC (p<0.05). MEKK3 can be used as an important marker of early diagnosis and prognostic evaluation in RCCC. It may be associated with imbalance of anti-tumor immunity and overexpression of pERK. Expression of MEKK3 and pERK are significantly increased in RCCC, with protein expression and clinical stage acting as independent prognostic factors.

RNAi-mediated silencing of the Skp-2 gene causes inhibition of growth and induction of apoptosis in human renal carcinoma cells.

Renal cancer ranks one of the most frequent causes of cancer death in the world. S-phase kinase-associated protein 2 (SKP 2) is overexpressed in human tumors and has prognostic value in many cancers including renal cancer, indicating its potential as a therapeutic target. In this study, we investigated the therapeutic potential of Skp-2 in renal cancer using the technique of RNA silencing via short hairpin RNA (shRNA). Synthetic shRNA duplexes against Skp-2 were introduced to down-regulate the expression of Skp-2 in a highly malignant renal carcinoma cell line, ACHN. The results indicated that siRNA targeting of Skp-2 could lead to an efficient and specific inhibition of endogenous Skp-2 activity. Furthermore, we found that depletion of Skp-2 caused a dramatic cell cycle arrest, followed by massive apoptotic cell death, and eventually resulted in a significant decrease in growth, viability and tumor formation in renal cancer cell lines studied.

Primary microcephaly gene MCPH1 shows a novel molecular biomarker of human renal carcinoma and is regulated by miR-27a.

Microcephalin 1 (MCPH1) gene, initially identified as an hTERT repressor, result in two autosomal recessive disorders: primary microcephaly and premature chromosome condensation syndrome. Recently, several studies have found that MCPH1 has also been shown to be downregulated in several different types of human cancers, suggesting that it could also function as a tumor suppressor gene and a novel molecular biomarker of human cancers. To investigate its potential role in the human renal carcinoma progression, we evaluated the expression of protein MCPH1 in 188 renal cancer and 20 normal renal tissues from 188 patients with renal cancer and 20 healthy persons by immunohistochemistry. Positive MCPH1 staining was found in all normal renal samples and partly in cancerous tissues. But MCPH1-positive cells resulted significantly lower in renal carcinoma tissues compared with normal tissues. We further observed that overexpression of MCPH1 decreased cellular proliferation, cell migration and invasion and induced cell apoptosis, indicating it is tumor suppressor. Using bioinformatics approaches and luciferase reporter assay, we showed that the 3'-UTR of MCPH1 harbors two non-overlapping functional seed regions for miR-27 which negatively regulated its level. The expression level of miR-27a negatively correlated with the MCPH1 protein level in renal cancer. Our study indicates for the first time that, in addition to its role in brain development, MCPH1 also functions as a tumor suppressor gene and is directly regulated by miR-27a.