Unveiling the Role of Protein Kinase C θ in Porcine Epidemic Diarrhea Virus Replication: Insights from Genome-Wide CRISPR/Cas9 Library Screening.

Porcine epidemic diarrhea virus (PEDV), a member of the Alpha-coronavirus genus in the Coronaviridae family, induces acute diarrhea, vomiting, and dehydration in neonatal piglets. This study aimed to investigate the genetic dependencies of PEDV and identify potential therapeutic targets by using a single-guide RNA (sgRNA) lentiviral library to screen host factors required for PEDV infection. Protein kinase C θ (PKCθ), a calcium-independent member of the PKC family localized in the cell membrane, was found to be a crucial host factor in PEDV infection. The investigation of PEDV infection was limited in Vero and porcine epithelial cell-jejunum 2 (IPEC-J2) due to defective interferon production in Vero and the poor replication of PEDV in IPEC-J2. Therefore, identifying suitable cells for PEDV investigation is crucial. The findings of this study reveal that human embryonic kidney (HEK) 293T and L929 cells, but not Vero and IPEC-J2 cells, were suitable for investigating PEDV infection. PKCθ played a significant role in endocytosis and the replication of PEDV, and PEDV regulated the expression and phosphorylation of PKCθ. Apoptosis was found to be involved in PEDV replication, as the virus activated the PKCθ-B-cell lymphoma 2 (BCL-2) ovarian killer (BOK) axis in HEK293T and L929 cells to increase viral endocytosis and replication via mitochondrial apoptosis. This study demonstrated the suitability of HEK293T and L929 cells for investigating PEDV infection and identified PKCθ as a host factor essential for PEDV infection. These findings provide valuable insights for the development of strategies and drug targets for PEDV infection.

The Combination of Anti-CD47 Antibody with CTLA4 Blockade Enhances Anti-Tumor Immunity in Non-Small Cell Lung Cancer via Normalization of Tumor Vasculature and Reprogramming of the Immune Microenvironment.

In solid tumors, the formidable anti-tumor impact resulting from blocking the "don't eat me" signal, arising from CD47-SIRPα interaction, is constrained, especially compared to its efficacy in hematopoietic malignancies. Activating macrophage anti-tumor activity not only necessitates the inhibition of the "don't eat me" signal, but also the activation of the "eat me" (pre-phagocyte) signal. Intriguingly, the cytotoxic T-lymphocyte-associated antigen 4 (CTLA4) antibody (Ab) has been identified to stimulate Fc receptor-mediated active phagocytes in the tumor microenvironment, thereby generating "eat me" signals. This study postulates that concurrently targeting CD47 and CTLA4 could intensify the anti-tumor effects by simultaneously blocking the "don't eat me" signal while triggering the "eat me" signal. The experimental data from this investigation confirm that the combined targeting of CD47 and CTLA4 enhances immunity against solid tumors in LLC cell-transplanted tumor-bearing mice. This effect is achieved by reducing myeloid-derived suppressor cell infiltration while increasing the presence of effector memory CD8+ T cells, NK1.1+ CD8+ T cells, and activated natural killer T cells. Meanwhile, combination therapy also alleviated anemia. Mechanistically, the anti-CD47 Ab is shown to upregulate CTLA4 levels in NSCLC cells by regulating Foxp1. Furthermore, targeting CD47 is demonstrated to promote tumor vascular normalization through the heightened infiltration of CD4+ T cells. These findings suggest that the dual targeting of CD47 and CTLA4 exerts anti-tumor effects by orchestrating the "eat me" and "don't eat me" signals, reshaping the immune microenvironment, and fostering tumor vascular normalization. This combined therapeutic approach emerges as a potent strategy for effectively treating solid tumors.

Overlapping cytoplasms segmentation via constrained multi-shape evolution for cervical cancer screening.

Segmenting overlapping cytoplasms in cervical smear images is a clinically essential task for quantitatively measuring cell-level features to screen cervical cancer This task, however, remains rather challenging, mainly due to the deficiency of intensity (or color) information in the overlapping region Although shape prior-based models that compensate intensity deficiency by introducing prior shape information about cytoplasm are firmly established, they often yield visually implausible results, as they model shape priors only by limited shape hypotheses about cytoplasm, exploit cytoplasm-level shape priors alone, and impose no shape constraint on the resulting shape of the cytoplasm In this paper, we present an effective shape prior-based approach, called constrained multi-shape evolution, that segments all overlapping cytoplasms in the clump simultaneously by jointly evolving each cytoplasm's shape guided by the modeled shape priors We model local shape priors (cytoplasm-level) by an infinitely large shape hypothesis set which contains all possible shapes of the cytoplasm In the shape evolution, we compensate intensity deficiency for the segmentation by introducing not only the modeled local shape priors but also global shape priors (clump-level) modeled by considering mutual shape constraints of cytoplasms in the clump We also constrain the resulting shape in each evolution to be in the built shape hypothesis set for further reducing implausible segmentation results We evaluated the proposed method in two typical cervical smear datasets, and the extensive experimental results confirm its effectiveness.

Significance of the cGAS-STING Pathway in Health and Disease.

The cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) pathway plays a significant role in health and disease. In this pathway, cGAS, one of the major cytosolic DNA sensors in mammalian cells, regulates innate immunity and the STING-dependent production of pro-inflammatory cytokines, including type-I interferon. Moreover, the cGAS-STING pathway is integral to other cellular processes, such as cell death, cell senescence, and autophagy. Activation of the cGAS-STING pathway by "self" DNA is also attributed to various infectious diseases and autoimmune or inflammatory conditions. In addition, the cGAS-STING pathway activation functions as a link between innate and adaptive immunity, leading to the inhibition or facilitation of tumorigenesis; therefore, research targeting this pathway can provide novel clues for clinical applications to treat infectious, inflammatory, and autoimmune diseases and even cancer. In this review, we focus on the cGAS-STING pathway and its corresponding cellular and molecular mechanisms in health and disease.

Vaccine induced memory CD8+ T cells efficiently prevent viral transmission from the respiratory tract.

Introduction: Mucosal immunization eliciting local T-cell memory has been suggested for improved protection against respiratory infections caused by viral variants evading pre-existing antibodies. However, it remains unclear whether T-cell targeted vaccines suffice for prevention of viral transmission and to which extent local immunity is important in this context. Methods: To study the impact of T-cell vaccination on the course of viral respiratory infection and in particular the capacity to inhibit viral transmission, we used a mouse model involving natural murine parainfluenza infection with a luciferase encoding virus and an adenovirus based nucleoprotein targeting vaccine. Results and discussion: Prior intranasal immunization inducing strong mucosal CD8+ T cell immunity provided an almost immediate shut-down of the incipient infection and completely inhibited contact based viral spreading. If this first line of defense did not operate, as in parentally immunized mice, recirculating T cells participated in accelerated viral control that reduced the intensity of inter-individual transmission. These observations underscore the importance of pursuing the development of mucosal T-cell inducing vaccines for optimal protection of the individual and inhibition of inter-individual transmission (herd immunity), while at the same time explain why induction of a strong systemic T-cell response may still impact viral transmission.

[Link between sortase A function and cariogenicity of Streptococcus mutans: a preliminary metabolomics analysis].

OBJECTIVE: This study intends to explore the mechanism underlying the support of sortase A (SrtA) of the cariogenicity of Streptococcus mutans (S. mutans). METHODS: We performed a metabonomics study based on ¹H nuclear magnetic resonance spectroscopy (NMR), in which we compared the extracellular metabolites of wild-type S. mutans UA159 with those of its SrtA-deficient strain. Metabolite differences among strains were identified using a combination of principal component analysis and orthogonality partial least square discriminant analysis. RESULTS: Several differences corresponding mostly to unknown metabolites were identified. Some amino acids such as leucine and valine (δ 0.92×10⁻6-1.20×10⁻6), lactic acid ( δ1.28×10⁻6), oxoglutaric acid (δ 3.00×10⁻6), and glycine (δ 3.60×10⁻6) differed among strains. CONCLUSIONS: This work establishes the feasibility of using ¹H NMR-based metabonomics to provide leads for research into molecular factors that promote caries. The database of microbial metabolites should be also improved in further studies.

The Pleiotropic Effects of PPARs on Vascular Cells and Angiogenesis: Implications for Tissue Engineering.

Angiogenesis is a complex process in which capillaries are produced from blood vessels that already exists. Endothelial cells (ECs) and endothelial progenitor cells (EPCs) are pivotal for this process and for the maintenance/restorage of the endothelium. Decreased numbers and dysfunction of these cells have been related to growing cardiovascular risks. Peroxisome-proliferator-activated receptor (PPAR) is a large family of nuclear receptors, characterized by three isotypes: α, ß and γ. Numerous studies have shown that PPAR activation is involved in the pathology of a wide range of cardiovascular diseases and has a role in endothelial function, thrombosis and inflammation, etc., suggesting that PPAR agonists may be good candidates to treat the cardiovascular disease. However, controversial results exist on whether this nuclear receptor is inductive or depressive in the process of angiogenesis. Herein, this review will provide a detailed discussion of the up-to-date investigation of the role of PPARs in angiogenesis, with particular reference to their effects on angiogenesis-related cells--i.e., ECs, EPCs, vascular smooth-muscle cells (VSMCs), macrophages and endometrial cells--and will discuss the current and potential future applications of PPAR activators.

Analysis of plasma metabolic biomarkers in the development of 4-nitroquinoline-1-oxide-induced oral carcinogenesis in rats.

The aim of the present study was to identify time-dependent changes in the expression of metabolic biomarkers during the various stages of oral carcinogenesis to provide an insight into the sequential mechanism of oral cancer development. An 1H nuclear magnetic resonance (NMR)-based metabolomics approach was used to analyze the blood plasma samples of Sprague-Dawley rats exhibiting various oral lesions induced by the administration of 4-nitroquinoline-1-oxide (4NQO) in drinking water. The 1H NMR spectra were processed by principal component analysis (PCA) and partial least-squares discriminant analysis (PLS-DA) to determine the metabolic differences between the three developmental stages of oral mucosa cancer (health, oral leukoplakia [OLK] and oral squamous cell carcinoma [OSCC]). The variable importance in projection (VIP) score derived from the PLS-DA model was used to screen for important metabolites, whose significance was further verified through analysis of variance (ANOVA). Data from the present study indicated that 4NQO-induced rat oral carcinogenesis produced oral pre-neoplastic and neoplastic lesions and provided an effective model for analyzing sequential changes in the 1H NMR spectra of rat blood plasma. The 1H NMR-based metabolomics approach clearly differentiates between healthy, OLK and OSSC rats in the PCA and PLS-DA models. Furthermore, lactic acid, choline, glucose, proline, valine, isoleucine, aspartic acid and 2-hydroxybutyric acid demonstrated VIP>1 in the PLS-D model and P<0.05 with ANOVA. It was also identified that increases in lactic acid, choline and glucose, and decreases in proline, valine, isoleucine, aspartic acid and 2-hydroxybutyric acid may be relative to the characteristic mechanisms of oral carcinogenesis. Therefore, these plasma metabolites may serve as metabolic biomarkers in oral carcinogenesis and assist in the early diagnosis and preventive treatment of oral cancer.

1H NMR-based metabonomic and pattern recognition analysis for detection of oral squamous cell carcinoma.

BACKGROUND: Metabonomic analysis has been increasingly used to monitor metabolic abnormalities in cells and their microenvironment in order to detect the cancer markers recently. We evaluated the feasibility of applying (1)H nuclear magnetic resonance ((1)H NMR) based metabonomic method in the early detection of the differences in the plasma from 3 groups, which were patients with oral squamous cell carcinoma (OSCC), patients with oral leukoplakia (OLK), and the healthy control group. METHODS: (1)H NMR spectra were obtained from human plasma samples prior to spectral analysis. The obtained data were processed by both the unsupervised principal component analysis (PCA) and the supervised partial least squares discriminant analysis (PLS-DA) to find out the differences among the three groups. RESULTS: PLS-DA analysis has revealed a good model to detect the NMR data that can differentiate the OSCC patients from the OLK patients and the controls using a test set. CONCLUSION: The results indicated that the (1)H NMR-based metabonomic approach is a feasible and efficient method for differentiating the OSCC patient's plasma from the healthy controls. As a potential novel strategy and a more convenient technique, it deserves a further evaluation for an early detection of oral cancer.

[The experimental study on biodegradable gel membrane applied to prevention of postoperative adhesion of abdominal cavity surgery].

OBJECTIVE: To develop a new type of anti-adhesion gel membrane and explore its applying technique. METHODS: 24 adult New Zealand white rabbits were used for the experiment research project, the animals were divided into two groups: the experiment group (18 adult New Zealand white rabbits) and the control group (6 adult New Zealand white rabbits). The animal models were established via the abdominal cavity. The biodegradable gel membrane was covered to the surface of the operating region in experimental animal group, while the 0.9% NaCL was directly rinsing the operating region in the controlled group. The specimens were collected at postoperatively week 2, 4 or 8 respectively. The samples were evaluated by global and histological observation. RESULTS: In the experiment group, a few adhed zones were observed in 2 and 4 weeks after operation, but in 8 weeks after operation, the adhesion zone was disappeared. In the control group, a few adheol zones were founded in 2 weeks after operation. In 4 weeks after operation, the adhed zone become conspicuously emerged. In 8 weeks after operation, the cicatri band was occurred. CONCLUSION: The biodegradable gel membrane usd into the postoperative abdominal cavity is effective and easy to manipulate, and it could be used as a high-effect, cheap drug of anti-adhesion in operation for surgeons.