IL-12-Overexpressed Nanoparticles Suppress the Proliferation of Melanoma Through Inducing ICD and Activating DC, CD8+ T, and CD4+ T Cells.

Purpose: The drug resistance and low response rates of immunotherapy limit its application. This study aimed to construct a new nanoparticle (CaCO3-polydopamine-polyethylenimine, CPP) to effectively deliver interleukin-12 (IL-12) and suppress cancer progress through immunotherapy. Methods: The size distribution of CPP and its zeta potential were measured using a Malvern Zetasizer Nano-ZS90. The morphology and electrophoresis tentative delay of CPP were analyzed using a JEM-1400 transmission electron microscope and an ultraviolet spectrophotometer, respectively. Cell proliferation was analyzed by MTT assay. Proteins were analyzed by Western blot. IL-12 and HMGB1 levels were estimated by ELISA kits. Live/dead staining assay was performed using a Calcein-AM/PI kit. ATP production was detected using an ATP assay kit. The xenografts in vivo were estimated in C57BL/6 mice. The levels of CD80+/CD86+, CD3+/CD4+ and CD3+/CD8+ were analyzed by flow cytometry. Results: CPP could effectively express EGFP or IL-12 and increase ROS levels. Laser treatment promoted CPP-IL-12 induced the number of dead or apoptotic cell. CPP-IL-12 and laser could further enhance CALR levels and extracellular HMGB1 levels and decrease intracellular HMGB1 and ATP levels, indicating that it may induce immunogenic cell death (ICD). The tumors and weights of xenografts in CPP-IL-12 or laser-treated mice were significantly reduced than in controls. The IL-12 expression, the CD80+/CD86+ expression of DC from lymph glands, and the number of CD3+/CD8+T or CD3+/CD4+T cells from the spleen increased in CPP-IL-12-treated or laser-treated xenografts compared with controls. The levels of granzyme B, IFN-γ, and TNF-α in the serum of CPP-IL-12-treated mice increased. Interestingly, CPP-IL-12 treatment in local xenografts in the back of mice could effectively inhibit the growth of the distant untreated tumor. Conclusion: The novel CPP-IL-12 could overexpress IL-12 in melanoma cells and achieve immunotherapy to melanoma through inducing ICD, activating CD4+ T cell, and enhancing the function of tumor-reactive CD8+ T cells.

m6A-methylated KCTD21-AS1 regulates macrophage phagocytosis through CD47 and cell autophagy through TIPR.

Blocking immune checkpoint CD47/SIRPα is a useful strategy to engineer macrophages for cancer immunotherapy. However, the roles of CD47-related noncoding RNA in regulating macrophage phagocytosis for lung cancer therapy remain unclear. This study aims to investigate the effects of long noncoding RNA (lncRNA) on the phagocytosis of macrophage via CD47 and the proliferation of non-small cell lung cancer (NSCLC) via TIPRL. Our results demonstrate that lncRNA KCTD21-AS1 increases in NSCLC tissues and is associated with poor survival of patients. KCTD21-AS1 and its m6A modification by Mettl14 promote NSCLC cell proliferation. miR-519d-5p gain suppresses the proliferation and metastasis of NSCLC cells by regulating CD47 and TIPRL. Through ceRNA with miR-519d-5p, KCTD21-AS1 regulates the expression of CD47 and TIPRL, which further regulates macrophage phagocytosis and cancer cell autophagy. Low miR-519d-5p in patients with NSCLC corresponds with poor survival. High TIPRL or CD47 levels in patients with NSCLC corresponds with poor survival. In conclusion, we demonstrate that KCTD21-AS1 and its m6A modification promote NSCLC cell proliferation, whereas miR-519d-5p inhibits this process by regulating CD47 and TIPRL expression, which further affects macrophage phagocytosis and cell autophagy. This study provides a strategy through miR-519-5p gain or KCTD21-AS1 depletion for NSCLC therapy by regulating CD47 and TIPRL.

Dimethyl Bisphenolate Ameliorates Carbon Tetrachloride-Induced Liver Injury by Regulating Oxidative Stress-Related Genes.

Liver disease accounts for millions of deaths per year all over the world due to complications from cirrhosis and liver injury. In this study, a novel compound, dimethyl bisphenolate (DMB), was synthesized to investigate its role in ameliorating carbon tetrachloride (CCl4)-induced liver injury through the regulation of oxidative stress-related genes. The structure of DMB was confirmed based on its hydrogen spectrum and mass spectrometry. DMB significantly reduced the high levels of ALT, AST, DBIL, TBIL, ALP, and LDH in a dose-dependent manner in the sera of CCl4-treated rats. The protective effects of DMB on biochemical indicators were similar to those of silymarin. The ROS fluorescence intensity increased in CCl4-treated cells but significantly weakened in DMB-treated cells compared with the controls. DMB significantly increased the content of oxidative stress-related GSH, Nrf2, and GCLC dose-dependently but reduced MDA levels in CCl4-treated cells or the liver tissues of CCl4-treated rats. Moreover, DMB treatment decreased the expression levels of P53 and Bax but increased those of Bcl2. In summary, DMB demonstrated protective effects on CCl4-induced liver injury by regulating oxidative stress-related genes.

Porcine reproductive and respiratory syndrome virus upregulates SMPDL3B to promote viral replication by modulating lipid metabolism.

Porcine reproductive and respiratory syndrome virus (PRRSV) poses a severe threat to the health of pigs globally. Host factors play a critical role in PRRSV replication. Using PRRSV as a model for genome-scale CRISPR knockout (KO) screening, we identified a host factor critical to PRRSV infection: sphingomyelin phosphodiesterase acid-like 3B (SMPDL3B). Our findings show that SMPDL3B restricted PRRSV attachment, entry, replication, and secretion and that its depletion significantly inhibited PRRSV proliferation, indicating that SMPDL3B plays a positive role in PRRSV replication. Our data also show that SMPDL3B deficiency resulted in an accumulation of intracellular lipid droplets (LDs). The expression level of key genes (ACC, SCD-1, and FASN) involved in lipogenesis was increased, whereas the fundamental lipolysis gene, ATGL, was inhibited when SMPDL3B was knocked down. Overall, our findings suggest that SMPDL3B deficiency can effectively inhibit viral infection through the modulation of lipid metabolism.

Predictive value of LDL/HDL ratio in coronary atherosclerotic heart disease.

BACKGROUND: Dyslipidemia is one of independent risk factors for coronary atherosclerotic heart disease (CAHD). We determined whether the LDL/HDL ratio is better than LDL-C or HDL-C alone in predicting the severity of CAHD. METHODS: We performed a retrospective study of 1351 patients with myocardial ischemia who underwent coronary angiography between January 2018 and December 2019 in Shanghai Ninth People's Hospital. Spearman correlation analysis, logistic regression model, Cox proportional hazards model and multicollinearity were used to evaluate LDL/HDL ratio for predicting CAHD severity compared to LDL-C or HDL-C alone. RESULTS: Higher LDL/HDL ratio was seen in CAHD patients than controls (2.94 ± 1.06 vs 2.36 ± 0.78, P < 0.05). LDL/HDL ratio was significantly associated with the severity of coronary vascular stenosis. The area under the ROC curve of LDL-C, HDL-C, LDL/HDL ratio used to predict CAHD are 0.574 (95% CI 0.547-0.600, P < 0.001), 0.625 (95% CI 0.598-0.651, P < 0.001), 0.668 (95% CI 0.639-0.697, P = 0.000), respectively. The cut-off value of LDL/HDL ratio is 2.517, and the sensitivity and specificity are 65% and 61%, respectively. LDL/HDL ratio was related to the prevalence of CAHD and the odds ratio (OR) was 2.39 [95% confidence interval (CI) 1.698-2.593, P = 0.00] in multicollinearity regression model. CONCLUSION: LDL/HDL ratio may become a better predictor of CAHD severity, compared to LDL-C or HDL-C.

Salmonella Enteritidis Subunit Vaccine Candidate Based on SseB Protein Co-Delivered with Simvastatin as Adjuvant.

Salmonella enterica serovar Enteritidis (S. Enteritidis) is an important zoonotic pathogen that can lead to diarrhea and systemic infections in humans and mortality in animals. This is a major public health issue worldwide. Safe and effective vaccines are urgently needed to control and prevent Salmonella infection. Subunit vaccines are safe and provide targeted protection against Salmonella spp. Here, we developed and evaluated an S. Enteritidis subunit vaccine candidate, the rHis-SseB adjuvant with simvastatin. We amplified the SseB gene from S. Enteritidis C50041 genomic DNA and expressed the recombinant proteins rHis-SseB and rGST-SseB using the Escherichia coli system. Western blotting confirmed the immunoreactivity of recombinant proteins rHis-SseB and rGST-SseB with antisera against Salmonella Enteritidis C50041. In a mouse model of intramuscular vaccination, co-immunization with rHis-SseB and simvastatin significantly enhanced both the SseB-specific antibody titer in serum (humoral immune response) and splenic lymphocyte proliferation (cellular immune response). Co-immunization with rHis-SseB and simvastatin provided 60% protection against subsequent challenge with the S. Enteritidis C50041 strain and decreased bacterial colonization in the liver and spleen. These findings provide a basis for the development of an S. Enteritidis subunit vaccine.

Studies on the Morphology Effect on Catalytic Ability of a Single MnO2 Catalyst Particle with a Solid Nanopipette.

Investigating the catalytic ability of an individual catalyst particle helps to understand heterogeneity and can provide new insights into the synthesis of high-efficiency catalysts. Solid-state nanopores have become a promising tool for detecting single molecules/particles due to their high temporal and spatial resolution. Here, we report a nanopore-based strategy for the evaluation and comparison of a single MnO2 catalyst particle with different morphologies by monitoring the generated O2 bubbles from the catalytic decomposition of H2O2. The finite element simulation was introduced to account for the flow velocity and bubble-induced current variation in the nanopore. In particular, the differences in catalytic ability of spherical and cubic MnO2 have been studied by calculating the production rate and volume of O2. It demonstrates that the shape of a single MnO2 catalyst particle has a significant effect on its catalytic activity indeed.

Two-dimensional shape retrieval from the interferometric out-of-focus image of a nonspherical particle-Part I: theory.

Shape is one of the important characteristics of nonspherical particles. Herein, the two-dimensional shapes of several micrometer/millimeter-sized particles with a variety of geometrical forms are retrieved from simulated interferometric out-of-focus images using the Hybrid Input-Output algorithm. The particle concerned can be either a single particle or a complex particle with separate parts. The impact of the axial size of the three-dimensional particle on the retrieval of the two-dimensional shape (i.e., the projection of the particle on the image sensor) is analyzed, showing that an increase of the axial size increases the deviation of low frequencies in the interferometric out-of-focus image and eventually degrades the quality of the reconstructed shape. This study demonstrates the capability of the interferometric out-of-focus imaging technique on shape information retrieval for micrometer/millimeter-sized nonspherical particles.

To increase the incorporation efficiency of genetically encoding Nε-acetyllysine in recombinant protein.

Reversible Nε-lysine (Nε-Lys) acetylation is a dynamic post-translational modification. Genetic incorporation of Nε-acetyllysine (Nε-AcK) into the specific site of a protein is a powerful method for producing recombinant protein with acetylation and studying the functional role of protein acetylation. Because of the universal existence of deacetylase such as CobB in vivo, the acetyl group of Nε-AcK may be removed from recombinant protein. So in the process of incorporating acetyl lysine into protein, nicotinamide (NAM), a lysine deacetylase (KDAC) inhibitor, is needed to inhibit the KDAC activity and protect the acetyl group of Nε-acetyllysine incorporated from removal in vivo. In this study, we knocked out cobB gene from an E. coli strain and used it as host for incorporating of Nε-AcK into recombinant protein by expanding genetic code. Comparing with the addition of KDAC inhibitor, this is a novel method to protect the acetyl group of Nε-acetyllysine from deacetylation by using cobB knockout strain. Thus, this method provides a better way for protecting recombinant acetylated protein from deacetylation. By knocking out cobB gene from the strain of E. coli, we successfully produced homogeneous acetylated protein by expanding genetic code with a high effectiveness. This method will also provide a good reference for genetically introducing other varieties of modified amino acids into protein.

Genome-wide identification of aberrantly methylated promoters in ovarian tissue of prenatally androgenized rats.

OBJECTIVE: To identify aberrantly methylated candidate genes that are involved in the development of polycystic ovary syndrome (PCOS). DESIGN: Animal model. SETTING: University-affiliated laboratory. ANIMAL(S): Sprague-Dawley rats. INTERVENTION(S): The prenatally androgenized (PNA) rat model was established. Pregnant rats were treated with daily SC injections of T propionate during late gestation, and their female offspring were studied as adults. MAIN OUTCOME MEASURE(S): Serum glucose and hormone levels, ovary morphology and cell apoptosis, genome-wide CpG methylation, and expression of caspase-3 protein were measured. RESULT(S): In the PNA group, the levels of serum glucose, 17-hydroxyprogesterone, and T were significantly higher when compared with the control group. Ovarian morphology showed increased atretic follicles and cystic follicles. Using the MeDIP-chip approach, we identified 528 genes that were hypermethylated in PNA ovaries. Gene ontology analyses revealed that these genes are involved in a variety of reproductive development and biological processes. The methylation enrichments of Bcl2l1 and Scr5a1 observed in the PNA group by MeDIP-quantitative polymerase chain reaction assay were significantly higher than those obtained from the control group. Furthermore, the mRNA level of the Bcl2l1 gene was significantly decreased in the PNA group. The percentage of caspase-3-positive cells in the PNA group was obviously higher compared with the control group, by terminal deoxynucleotidyl transferase dUTP nick end labeling detection as well. CONCLUSION(S): DNA methylation alteration may be an important factor affecting the genes involved in the pathophysiological processes that result in the phenotype of PCOS.

Cylindrical interferometric out-of-focus imaging for the analysis of droplets in a volume.

We propose an original cylindrical interferometric out-of-focus imaging setup to realize the characterization of spherical droplets in a volume. The longitudinal position of the droplets is determined through the orientation of the fringes while the diameter of the droplets is obtained from the frequency of the fringes. The experiments agree with the simulations.

Interferometric laser imaging for droplet sizing revisited: elaboration of transfer matrix models for the description of complete systems.

We report the development of an interferometric laser imaging for droplet sizing (ILIDS) numerical simulator. It is based on the use of generalized Huygens-Fresnel integrals associated to transfer matrices that describe the whole imaging setup. This simulator allows easy simulation of any kind of ILIDS setup. Simulations are shown to be in good agreement with experimental results. This simulator offers important perspectives in the design, realization, and calibration of ILIDS instruments, as airborne instruments, or in situ measurements in flows.

Extended ABCD matrix formalism for the description of femtosecond diffraction patterns; application to femtosecond digital in-line holography with anamorphic optical systems.

We present a new model to predict diffraction patterns of femtosecond pulses through complex optical systems. The model is based on the extension of an ABCD matrix formalism combined with generalized Huygens-Fresnel transforms (already used in the CW regime) to the femtosecond regime. The model is tested to describe femtosecond digital in-line holography experiments realized in situ through a cylindrical Plexiglas pipe. The model allows us to establish analytical relations that link the holographic reconstruction process to the experimental parameters of the pipe and of the incident beam itself. Simulations and experimental results are in good concordance. Femtosecond digital in-line holography is shown to allow significant coherent noise reduction, and this model will be particularly efficient to describe a wide range of optical geometries. More generally, the model developed can be easily used in any experiment where the knowledge of the precise evolution of femtosecond transverse patterns is required.