An Engineered DC-Targeting Lentivector Induces Robust T Cell Responses and Inhibits HBV Replication in HBV Transgenic Mice via Upregulating T Cell Autophagy

Background/Aims: Developing engineered dendritic cell (DC)-targeting lentivectors (LVs) have been the target of intense research for their potential to create antigen-directed immunotherapeutics which can be safely administered to patients. In this study, we constructed a DC-directed LV (LVDC-UbHBcAg-LIGHT) as a potential vaccine to induce anti-HBV immune responses. Methods: Specificity of LVDC-UbHBcAg-LIGHT for DCs in vivo was confirmed through live animal imaging studies. The levels of cytokine production in T cells were assessed by flow cytometry. The HBcAg-specific cytotoxic T lymphocyte (CTL) responses and antibody responses induced by direct administration of the LVs were detected by LDH release assay and ELISA respectively. The levels of serum HBsAg and HBV DNA were evaluated by Abbott kits and quantitative PCR respectively. The expression levels of HBsAg and HBcAg in liver tissues of HBV transgenic mice were examined by immunohistochemistry. In addition, molecular mechanism underlying the activation of CD8+ T cells was explored. Results: Live animal imaging studies showed that following subcutaneous administration of LVDC-UbHBcAg-LIGHT, no obvious luminescence signal was detected at the injection site. Immunization with LVDC-UbHBcAg-LIGHT elicited potent T cell responses in HBV transgenic mice evidenced by increased percentages of IFN-γ, TNF-α and GzmB producing CD8+ T cells as well as IFN-γ producing CD4+ T cells, improved HBcAg-specific CTL activities and antibody responses. Additionally, vaccination with LVDC-UbHBcAg-LIGHT efficiently reduced serum HBsAg, HBV DNA levels and the expression of HBsAg and HBcAg in liver tissues of HBV transgenic mice. More importantly, autophagy was induced in the activated CD8+ T cells, and the induced autophagy noticeably promoted the proliferation of T cells and decreased the frequencies of apoptotic CD8+ T cells by selectively degrading ubiquitinated apoptosis and cell cycle-associated protein aggregates. Futhermore, we confirmed the interaction between autophagosomes and ubiquitinated aggregates by confocal microscopy and immunoprecipitation analysis. Conclusions: These results demonstrated that LVDC-UbHBcAg-LIGHT provided a simple method of eliciting effective antiviral immune responses in HBV transgenic mice and might potentially be used as a therapeutic strategy to eradicate HBV with more safety and efficiency. Moreover, our results revealed a direct role of autophagy in promoting the survival and proliferation of activated CD8+ T cells

MACROD1/LRP16 Enhances LPS-Stimulated Inflammatory Responses by Up-Regulating a Rac1-Dependent Pathway in Adipocytes

Background/Aims: Chronic inflammation contributes to the development of type 2 diabetes mellitus by targeting the insulin receptor substrate protein-1 (IRS-1) signaling pathway. Previous studies showed that Leukemia related protein 16 (LRP16) reduced insulin stimulated glucose uptake in adipocytes by impairing the IRS-1 signaling pathway. We explored the mechanism by which LRP16 promotes the inflammatory response. Methods: We screened LRP16 induced proteins in the lipopolysaccharide (LPS)-stimulated inflammatory response using liquid chromatography-mass spectrometry (LC-MS) and analyzed the potential biological functions of these proteins using online bioinformatics tools. mRNA expression and protein expression of target genes were measured by real time PCR and Western blot, respectively. Results: A total of 390 differentially expressed proteins were identified. The mitogen-activated protein kinase (MAPK) signaling pathway was the primary activated pathway in LRP16-expressing cells. Overexpression of LRP16 activated ERK1/2 and Rac1, which are two key players related to the MAPK signaling pathway. Furthermore, knock down of endogenous LRP16 by RNA interference (RNAi) reduced Rac1 expression, ERK activation, and inflammatory cytokine expression in human adipocytes stimulated by LPS. The stimulatory effect of LRP16 was diminished by suppressing Rac1 expression and treating the cells with the ERK specific inhibitor, PD98059. Conclusion: These findings revealed the functions of LRP16 in promoting the inflammatory response through activating the Rac1-MAPK1/ERK pathway in human adipocytes

Diagnostic Role of Prostate-Specific Membrane Antigen in Adrenocortical Carcinoma

Objective: To investigate the role of PSMA in the differential diagnosis of adrenocortical carcinoma samples (ACCs) and adrenocortical adenoma samples (ACAs), to validate the prognostic role of PSMA in patients with ACCs, and to explore the possibility that this marker can differentiate localized ACCs from adrenal metastases from other sites.Methods: PSMA protein expression in tissue samples from 50 ACCs, 90 ACAs (including 20 from patients who presented with Cushing's syndrome, 20 aldosterone-producing adenomas and 50 non-functional tumors) and 10 tissues that were metastases from other primary sites was assessed by immunohistochemistry. The clinical and pathological characteristics were compared, the intensity and density were analyzed, and the prognostic role was evaluated.Results: The analysis of clinical and pathological features revealed that the size of ACCs was greater than that of benign tissues and the ACC patients were older than the ACA patients (p < 0.01). The percentage of PSMA-positive vessels, the mean intensity and the degree of staining density were found to be significantly lower in ACAs than in ACCs (p < 0.01). In these 140 samples, 60% of the ACCs were grouped in the positive category. The samples were negative for metastases that were from other primary sites. The ENSAT stage and Ki-67 were correlated with PSMA expression. The survival distribution revealed that high PSMA expression did not show any prognostic relevance in the current ACCs series. Those samples with a score of > 3.5 were 75 times more likely to be malignant (OR = 75). We established a cut-off score of 3.5 (p < 0.05), which had 46% sensitivity and 99% specificity. Paralleling PSMA and Ki-67 maximized the area under the curve, with 72% sensitivity and 100% specificity.Conclusions: Our results strongly confirm that PSMA is helpful for distinguishing benign from malignant tumors and that its high expression levels correlate with a high ENSAT stage and high proliferation. The combination of PSMA and Ki-67 can be particularly useful. Furthermore, PSMA might be a useful tool for the identification of localized adrenal carcinoma and metastatic carcinoma

Polymerizable Surfactant Ligand for Stabilization and Film Formation of CsPbBr₃ Nanocrystals

Surfactant ligands are important in the synthesis of inorganic perovskite nanocrystals (NCs), not only for stabilizing NCs but also for surface defect passivation. A new polymerizable surfactant ligand with a multidentate l-cysteine head, a long oleoyl tail, and a polymerizable styrenyl group (NOSVC) is designed for the post-synthesis treatment and stabilization of colloidal CsPbBr3 NCs in this work. 1H nuclear magnetic resonance and X-ray photoelectron spectroscopy analysis show that the l-cysteine head has strong interactions with the NCs. The absolute photoluminescence quantum yields of the colloidal NCs are increased from 45.1% of the pristine NCs stabilized with oleic acid/oleyl amine to 91.8% after NOSVC treatment. NOSVC-stabilized CsPbBr3 colloidal NCs show enhanced stabilities when exposed in polar solvents. The NOSVC-stabilized CsPbBr3 NCs in a solid film state allow for a photopolymerization to be carried out with the assistance of a photoinitiator. The polymerized films of NOSVC-treated NCs exhibit significantly enhanced stability against thermal radiation, ultraviolet irradiation, and humidity. We also fabricated self-healing polymer films incorporating NOSVC-treated CsPbBr3 NCs as a green filter for a white light-emitting diode device. The green light-emitting films are very stable in humid environments, revealing the great application potential of NOSVC-treated CsPbBr3 NCs in flexible display and lighting devices

The design and experiment of peanut high-throughput automatic seed testing system based on machine learning

In view of the fact that the current peanut test was still artificial, it was difficult to meet the actual demand of peanut test, this paper established a peanut high-throughput automatic test system (automatic measurement), which improves the work efficiency of peanut test. The system mainly included peanut automatic seed testing device, scanning gun, industrial computer, multi-serial controller and peanut automatic seed testing system, which could obtain the automation of the whole process of peanut pod testing and kernel fruit testing in real time. The results showed that the system can measure the peanut pods and nuts and other parameters in real time, and the average measuring accuracy of the length, width, diameter and quantity of the peanut pod and kernel was more than 98%. Compared with the manual test, the automatic measurement can significantly improved the test efficiency. The system was also suitable for the acquisition of other crop test parameters and provides a reference for high-throughput automatic test

TEM Investigation of Asymmetric Deposition-Driven Crystalline-to-Amorphous Transition in Silicon Nanowires

Controlling the shape and internal strain of nanowires (NWs) is critical for their safe and reliable use and for the exploration of novel functionalities of nanodevices. In this work, transmission electron microscopy was employed to examine bent Si NWs prepared by asymmetric electron-beam evaporation. The asymmetric deposition of Cr caused the formation of nanosized amorphous-Si domains; the non-crystallinity of the Si NWs was controlled by the bending radius. No other intermediate crystalline phase was present during the crystalline-to-amorphous transition, indicating a direct phase transition from the original crystalline phase to the amorphous phase. Moreover, amorphous microstructures caused by compressive stress, such as amorphous Cr domains and boxes, were also observed in the asymmetric Cr layer used to induce bending, and the local non-crystallinity of Cr was lower than that of Si under the same bending radius