Using 1/2 Descending Time in CEUS to Identify Renal Allograft Rejection.

RATIONALE AND OBJECTIVES: This study investigates the potential of quantitative Contrast-Enhanced Ultrasound (CEUS) parameters to distinguish between graft dysfunction due to rejection and non-rejection in kidney transplant recipients. METHODS: In this retrospective study, 50 kidney transplant patients who presented elevated serum creatinine or proteinuria were analyzed. They were categorized as rejection or non-rejection based on biopsy outcomes. These classifications were applied in both derivation (n = 33) and validation cohorts (n = 17). Prior to the biopsy, all patients underwent a CEUS. Quantitative parameters derived from the CEUS were further analyzed for their consistency and reliability. Additionally, the relationship between the Banff scores, a standard for diagnosing transplant rejections, and these CEUS parameters was explored. RESULTS: Significant differences between rejection and non-rejection groups were observed in the CEUS parameters of derivation cohorts. Specifically, Peak Intensity (PI), 1/2 Descending Time (DT/2), Area Under Curve (AUC), and Mean Transit Time (MTT) stood out. Sensitivity and specificity for these parameters were 76.5% and 87.5% for PI, 76.5% and 81.2% for DT/2, 76.5% and 87.5% for AUC, and 68.8% and 94.1% for MTT, respectively. DT/2 and MTT showed superior interobserver agreement compared to PI and AUC. When extrapolating the cutoff values from the derivation cohort to the validation group, DT/2 and AUC exhibited optimal diagnostic precision with positive and negative predictive values being 91.7% vs. 100% and 100% vs. 85.7%, respectively. Additionally, DT/2 effectively differentiated between mild and moderate to severe microvascular inflammation, pivotal in diagnosing antibody-mediated renal transplant rejection. CONCLUSION: DT/2 from CEUS parameters presents as a reliable tool to differentiate rejection from non-rejection causes in renal transplant dysfunction. Yet, large-scale, multi-center studies are essential for further validation.

ISX-9 Promotes KGF Secretion From MSCs to Alleviate ALI Through NGFR-ERK-TAU-ß-Catenin Signaling Axis.

BACKGROUND: Mesenchymal stem cells (MSCs) have been widely studied to alleviate acute lung injury (ALI) due to their paracrine function. However, the microenvironment of inflammatory outbreaks significantly restricted the factors secreted from MSCs like keratinocyte growth factor (KGF). KGF is a growth factor with tissue-repaired ability. Is there a better therapeutic prospect for MSCs in combination with compounds that promote their paracrine function? Through compound screening, we screened out isoxazole-9 (ISX-9) to promote MSCs derived KGF secretion and investigated the underlying mechanisms of action. METHODS: Compounds that promote KGF secretion were screened by a dual-luciferase reporter gene assay. The TMT isotope labeling quantitative technique was used to detect the differential proteins upon ISX-9 administrated to MSCs. The expressions of NGFR, ERK, TAU, and ß-catenin were detected by Western blot. In the ALI model, we measured the inflammatory changes by HE staining, SOD content detection, RT-qPCR, immunofluorescence, etc. The influence of ISX-9 on the residence time of MSCs transplantation was explored by optical in vivo imaging. RESULTS: We found out that ISX-9 can promote the expression of KGF in MSCs. ISX-9 acted on the membrane receptor protein NGFR, upregulated phosphorylation of downstream signaling proteins ERK and TAU, downregulated phosphorylation of ß-catenin, and accelerated ß-catenin into the nucleus to further increase the expression of KGF. In the ALI model, combined ISX-9 with MSCs treatments upgraded the expression of KGF in the lung, and enhanced the effect of MSCs in reducing inflammation and repairing lung damage compared with MSCs alone. CONCLUSIONS: ISX-9 facilitated the secretion of KGF from MSCs both in vivo and in vitro. The combination of ISX-9 with MSCs enhanced the paracrine function and anti-inflammatory effect of MSCs compared with MSCs applied alone in ALI. ISX-9 played a contributive role in the transplantation of MSCs for the treatment of ALI.

Evaluation of Microwave Ablation in 4T1 Breast Tumor by a Novel VEFGR2 Targeted Ultrasound Contrast Agents.

OBJECTIVES: A novel ultrasound contrast agent (UCA) VEGFR2-targeting iron-doped silica (SiO2) hollow nanoparticles (VEGFR2-PEG-HSNs-Fe NPs) was prepared and applied in microwave ablation for breast cancer to investigate its value in the evaluation of effectiveness after tumor ablation. METHODS: VEGFR2-PEG-HSNs-Fe NPs were prepared by using nano-SiO2, which was regarded as a substrate and etched by ferrous acetate, and then modified with anti-VEGFR2 antibody. Laser confocal microscope and flow cytometry were used to observe its main physicochemical properties, and biological safety was also investigated. After the xenograft tumor was treated with microwave ablation, the extent of perfusion defect was evaluated by ultrasound by injecting VEGFR2-PEG-HSNs-Fe NPs. RESULTS: The average particle size of VEGFR2-PEG-HSNs-Fe was 276.64 ± 30.31 nm, and the surface potential was -13.46 ± 2.83 mV. In vitro, the intensity of ultrasound signal increased with UCA concentration. Good biosafety was performed in in vivo and in vitro experiments. The enhanced ultrasound signal was detected in tumors after injection of VEGFR2-PEG-HSNs-Fe NPs, covering the whole tumor. The lesions, which were incompletely ablated, presented as contrast agent perfusion at the periphery of the tumor, and contrast enhanced ultrasound (CEUS) was performed again after complementary ablation. It was confirmed that all the lesions were completely ablated. CONCLUSION: Nano-targeted UCAs VEGFR2-PEG-HSNs-Fe NPs had good biosafety and ability of specific imaging, which might be used as a contrast agent in CEUS to evaluate the efficacy of tumor ablation.

The role of angiotensin-(1-7) on acquired platinum resistance-induced angiogenesis in non-small cell lung cancer in vitro and in vivo.

Renin-angiotensin system (RAS) signaling has been implicated in the development of cancer. The new RAS ACE2/Ang-(1-7)/Mas axis antagonizes the classical ACE/Ang II/AT1R axis. Ang-(1-7) has pleiotropic roles in lung cancer including suppressing proliferation, angiogenesis, and metastasis. This research was designed to investigate the effect of Ang-(1-7) on tumor-associated angiogenesis in DDP-resistant lung cancer cell lines. We first established acquired DDP-resistant cell lines A549 (A549-DDP) and LLC (LLC-DDP). We next performed RT-qPCR, western blot, ELISA, tube formation, microvessel density detection, immunohistochemistry, and tumor formation assays. The results showed that the mRNA and protein levels of RAS components and vascular endothelial growth factor A (VEGFa) were lessened in the A549/LLC-DDP+Ang-(1-7) group compared with the A549/LLC-DDP group. This effect could be blocked by the MAS receptor antagonist A779. The data revealed that Ang-(1-7) could perform its antiangiogenic function by PI3K/AKT and MAPK pathways. Furthermore, the impact of Ang-(1-7) on tumor-associated angiogenesis has been confirmed in lung cancer xenograft model with acquired DDP resistance. These results provide a theoretical basis for designing therapeutic strategies for targeting Ang-(1-7) in the treatment of NSCLC.

Analysis of liver steatosis analysis and controlled attenuation parameter for grading liver steatosis in patients with chronic hepatitis B.

BACKGROUND: Chronic hepatitis B is the most common chronic liver disease in China. For patients with chronic hepatitis B, steatosis increases the risk of cirrhosis and hepatocellular carcinoma. This study aimed to analyze and compare the clinical value of a newly developed ultrasound attenuation parameter, liver steatosis analysis (LiSA), acquired by Hepatus (Mindray, China), and controlled attenuation parameter (CAP), a widely used ultrasound attenuation parameter acquired by FibroScan (Echosens, France), for grading liver steatosis in patients with chronic hepatitis B infection. METHODS: A total of 203 patients were divided into two groups according to liver fat content validated by liver biopsy: group 1 (liver fat content <10%) and group 2 (liver fat content ≥10%). All patients underwent LiSA and CAP examinations. Receiver operating characteristic (ROC) curves were calculated for the two ultrasound attenuation tools. RESULTS: Both LiSA and CAP successfully discriminated between patients in group 1 and group 2. ROC curves showed that both tools had good diagnostic ability (AUC: >0.7) for steatosis ≥10%, and the performance of LiSA was significantly better than CAP (AUC: 0.859 vs. 0.801, P=0.048). Using optimal cut-off points, LiSA had specificity and sensitivity of 96.23% and 76.08%, respectively, for the diagnosis of steatosis ≥10%, compared to 91.53% and 72.10%, respectively, for CAP. CONCLUSIONS: LiSA and CAP are extremely efficient tools for assessing liver steatosis, even at a low grade. Both parameters are non-invasive, inexpensive, and easy to use, and can provide immediate results with high sensitivity.

A New Visual Transient Elastography Technique for Grading Liver Fibrosis in Patients With Chronic Hepatitis B.

ABSTRACT: Liver fibrosis is evaluated to assess the prognosis and guide the treatment of chronic hepatitis B (CHB). To compare the efficiency of 2 transient elastography techniques for grading liver fibrosis in CHB: visual transient elastography (ViTE) with real-time image guidance and FibroScan (FS) with no image guidance. All of the CHB patients in this study underwent both FS and ViTE examinations. The final diagnosis was based on the histological findings of a liver biopsy. According to the severity of liver fibrosis (based on the Scheuer criteria), the area under the receiver operating characteristic curve values for diagnostic efficiency were calculated for the 2 elastography techniques. This study enrolled 227 patients (79 [39.1%] women; mean age, 45.8 ± 16.8 years). The ViTE and FS liver elasticity measurements were highly correlated with liver fibrosis stage (r = 0.852 and r = 0.813, respectively). The area under the receiver operating characteristic curve value was larger for ViTE compared with FS, with respect to differentiating liver fibrosis stage, but not significantly (P > 0.05). The ViTE and FS can be used to detect and stage liver fibrosis. ViTE, easier and quicker to perform with superior interoperator reproducibility, is a stable and reliable elastography technique that benefits from real-time visual guidance.

Preparation of colloidal polydopamine/Au hollow spheres for enhanced ultrasound contrast imaging and photothermal therapy.

Development of functional theranostic platform for systemic contrast-enhanced diagnostic imaging and therapy is of great necessity in nanomedicine. However, synthesizing the biocompatible theranostic agents with enhanced merits in imaging and therapy via a facile and green way is still highly challenged. Here, we report a novel theranostic agent based on colloidal polydopamine/Au (PDA/Au) hollow spheres, which are synthesized with combined use of PDA chemistry and sacrificial template techniques. Colloidal polystyrene (PS) spheres serve as the templates with coatings of a PDA shell and Au nanoparticles in sequence, which are subsequently removed with trichloromethane, giving rise to colloidal PDA/Au hollow spheres. Colloidal PDA/Au hollow spheres exhibit excellent contrast enhancement for ultrasound imaging, and can serve as the near-infrared (NIR) photoabsorbers for the effective photothermal ablation of 4 T1 breast cancer cells in vitro with minor cytotoxicity to living cells. This method suggests a novel avenue for theranostic treatment in oncology.

Targeted Fe-doped silica nanoparticles as a novel ultrasound-magnetic resonance dual-mode imaging contrast agent for HER2-positive breast cancer.

BACKGROUND: Multimodal contrast agents with low toxicity and targeted modification have opened up new possibilities for specific imaging of breast cancer and shown broad application prospects in biomedicine and great potential for clinical transformation. In this work, a potential multifunctional imaging agent was developed by doping Fe into hollow silica nanoparticles (HS-Fe NPs), followed by modification with specific anti-HER2 antibodies, enabling the NPs to have dual-mode ultrasound (US)-magnetic resonance (MR)-specific imaging capacity with low toxicity. METHODS: Anti-HER2 antibodies were conjugated to silane-polyethylene glycol (PEG)-COOH-modified HS-Fe (HS-Fe-PEG) NPs to produce HER2-targeted HS-Fe-PEG (HS-Fe-PEG-HER2) NPs. The toxicity of HS-Fe-PEG-HER2 NPs on targeted cells in vitro and blood and organ tissue of mice in vivo was investigated. Distribution in vivo was also studied. Confocal laser-scanning microscopy and flow cytometry were used to evaluate the targeting ability of HS-Fe-PEG-HER2 NPs in vitro. US and MR instruments were used for imaging both in vivo and in vitro. RESULTS: The obtained HS-Fe-PEG-HER2 NPs (average diameter 234.42±48.76 nm) exhibited good physical properties and biosafety. In solution, they showed obvious enhancement of the US signal and negative contrast in T 2-weighted MR imaging. The binding rate of HS-Fe-PEG-HER2 NPs to targeted cells (SKBR3) was 78.97%±4.41% in vitro. US and MR imaging in vivo confirmed that the HS-Fe-PEG-HER2 NPs were delivered passively into the tumor region of SKBR3 and bound specifically to tumor cells. Target enhancement was better than untargeted and targeted competition groups. CONCLUSION: HS-Fe-PEG-HER2 NPs have potential as a low-cytotoxicity and dual-mode US-MR-specific imaging agent.

Identifying macrophage enrichment in atherosclerotic plaques by targeting dual-modal US imaging/MRI based on biodegradable Fe-doped hollow silica nanospheres conjugated with anti-CD68 antibody.

Macrophage recruitment has emerged as the crucial force driving the initiation and progression of atherosclerotic lesions. Therefore, the identification of macrophages in plaques is of vital importance for identifying vulnerable plaques, and noninvasive imaging methods are particularly desirable. Some studies have reported that MRI can detect plaque macrophages through targeted nanoparticles, but it is still hard for an US to detect macrophages in atherosclerotic plaque. In this study, anti-CD68 receptor-targeted Fe-doped hollow silica nanoparticles (CD68-Fe-HSNs) were fabricated as a dual-modal US/MRI contrast agent for identifying macrophages of aorta ventralis atherosclerotic plaques in ApoE-/- mice, confirmed by immunofluorescence and bio-TEM. This system possesses biodegradable characteristics even though it is an inorganic mesoporous nanosystem, indicating its potential high biocompatibility for further in vivo research. We expect that these dual-modal US/MRI nanoparticles will play a role in assessing vulnerable plaque in future research studies.

Twisted Gastrulation BMP Signaling Modulator 1 Regulates Papillary Thyroid Cancer Cell Motility and Proliferation.

Bone morphogenetic proteins (BMPs) are growth factors that have important functions in cell proliferation, migration and differentiation. To date, BMP pathway activation has been found in multiple human tumors and is associated with enhanced malignant tumor growth and metastasis. BMP activity is tightly regulated by a family of soluble extracellular secreted BMP modulators. Twisted gastrulation BMP signaling modulator 1 (TWSG1) is a direct BMP regulator that is required for the full signaling activity of BMPs. However, the functions and mechanisms of TWSG1 in papillary thyroid cancer (PTC) metastasis have not been reported. TWSG1 expression was detected in 44 PTC tissues with lymph node metastasis (LNM) and 56 PTC tissues without LNM using quantitative real-time polymerase chain reaction (qRT-PCR). Gain- and loss-of-function approaches were used to assess the biological function of TWSG1 in PTC cells. Matrigel assays demonstrated the effect of tumor cell-derived TWSG1 on endothelial cell function. Our results showed that TWSG1 expression was significantly enhanced in PTC with LNM compared to that in PTC without LNM. TWSG1 knockdown inhibited migration, invasion and proliferation of PTC cells. Additionally, TWSG1 suppression impaired the tumor cell-induced endothelial cell sprout formation. We found that TWSG1 signaling may be transduced by the BMP target transcription factor inhibitor of DNA binding 1 (Id1) and matrix metalloproteinases (MMPs) 2 and 9. In conclusion, TWSG1 was highly expressed in metastasized PTC; tumor growth, migration and invasion were dependent on TWSG1, and it may be a new diagnostic and therapeutic target for PTC.

Long noncoding RNA papillary thyroid carcinoma susceptibility candidate 3 (PTCSC3) inhibits proliferation and invasion of glioma cells by suppressing the Wnt/ß-catenin signaling pathway.

BACKGROUND: The dysregulation of long noncoding RNAs (lncRNAs) has been identified in a variety of cancers. An increasing number of studies have found the critical role of lncRNAs in the regulation of cellular processes, such as proliferation, invasion and differentiation. Long noncoding RNA papillary thyroid carcinoma susceptibility candidate 3 (PTCSC3) is a novel lncRNA that was primarily detected in papillary thyroid carcinoma. However, the biological function and molecular mechanism of lncRNA PTCSC3 in glioma are still unknown. METHODS: The expression level of lncRNA PTCSC3 in human microglia and glioma cell lines was examined using quantitative real-time polymerase chain reaction (qRT-PCR). The influence of lncRNA PTCSC3 on cell proliferation were studied using the cell counting kit-8, and cell cycle and apoptosis were analyzed by flow cytometry assays. The migration and invasion abilities were investigated by transwell and wound healing assays. The target genes of lncRNA PTCSC3 were explored by qRT-PCR, immunofluorescence and western blot. RESULTS: LncRNA PTCSC3 was significantly downregulated in glioma cell lines. The overexpression of lncRNA PTCSC3 suppressed proliferation and induced apoptosis in U87 and U251 cells. Additionally, the overexpression of lncRNA PTCSC3 inhibited the migration and invasion of U87 and U251 cells. Moreover, lncRNA PTCSC3 inhibited the epithelial-mesenchymal transition of U87 cells. The study also demonstrated that LRP6, as a receptor of the Wnt/ß-catenin pathway, was a target of lncRNA PTCSC3. By evaluating the expression levels of Axin1, active ß-catenin, c-myc, and cyclin D1, the study indicated that lncRNA PTCSC3 inhibited the activation of the Wnt/ß-cateninpathway through targeting LRP6. CONCLUSIONS: LncRNA PTCSC3 inhibits the proliferation and migration of glioma cells and suppresses Wnt/ß-catenin signaling pathway by targeting LRP6. LncRNA PTCSC3 is a potential therapeutic target for treatment of glioma.

Celecoxib regulates apoptosis and autophagy via the PI3K/Akt signaling pathway in SGC-7901 gastric cancer cells.

Gastric cancer, one of the most common malignancies worldwide, typically has a poor prognosis and poor survival rate. Previous studies have investigated the chemopreventive effect of celecoxib. In the present study, the SGC-7901 human gastric cancer cell line was utilized to examine the chemopreventive mechanisms of celecoxib. The inhibition of cell proliferation was determined using MTT assay, cell apoptosis was monitored by terminal deoxynucleotidyl transferase-mediated dUTP nick end­labeling (TUNEL) and flow cytometry, and cell ultrastructural changes were assessed via transmission electron microscopy. The mRNA expression of Akt, caspase-8 and -9 was examined using quantitative reverse-transcription-polymerase chain reaction (qRT-PCR) and p-Akt, procaspase-8 and -9 were analyzed via western blotting. The results showed that celecoxib inhibited the proliferation of SGC-7901 cells in a time- and dose-dependent manner. Additionally, celecoxib induced apoptosis as substantiated by typical apoptotic bodies, autophagosomes and an increased apoptotic rate. It was found that following celecoxib treatment, Akt mRNA expression was not significantly altered, and that p-Akt protein levels decreased in a time- and dose­dependent manner. Additionally, caspase-8 and -9 mRNA expression was significantly increased, while procaspase-8 and -9 protein expression decreased relative to the time- and dose-dependent effects. These results demonstrated that celecoxib induced apoptosis and autophagy of gastric cancer cells in vitro through the PI3K/Akt signaling pathway. Moreover, our findings suggested that celecoxib induces apoptosis in gastric cancer cells through the mitochondrial and death receptor pathways, providing additional understanding regarding the chemopreventive behaviors of celecoxib and its uses in cancer therapy.

[Clinical analysis of 87 cases of testicular tumor].

OBJECTIVE: To improve the diagnosis and treatment of testicular tumor. METHODS: Eighty-seven cases of testicular tumor were retrospectively studied. RESULTS: Of the total number, 79 cases were pathologically diagnosed as germ cell tumor (90.1%), among which there were 44 cases of seminoma (55.7%) and 7 cases of benign tumor (8.1%). Nonseminoma germ cell tumor (NSGCT) was found mainly among those under 5 and from 18 to 40 years of age, while seminoma chiefly among those beyond 17, and testis tumor was rare among those between 5 and 17 years old (1 case only). Three-year and 5-year survival rates of seminoma and NSGCT were 90.6% and 81.3%, and 83.3% and 56.7%, respectively. CONCLUSION: (1) Testicular tumors are mostly germ cell tumors. (2) NSGCT develops mainly among those under the age of 5 and from 18 to 40. (3) Seminoma is rare in those under 18. (4) Testicular tumor rarely develops among those between 5 and 17 years old. (5)Three-year and 5-year survival rates for seminoma are higher than those for NSGCT.