Blood Of Mice Research Articles

Anti-tumoral Immunity and Chemo-preventive Effectiveness of Herbal Extracts of Curcumin, Ginger, Clove and Amygdaline in Ehrlich Ascites Carcinoma-Challenging Mice.

Due to its systemic toxicity, traditional chemotherapy of tumors is being taken into consideration. Herbal therapy, containing phytochemical polyphenol derivatives such as Curcumin (Cur), Ginger (Gin), Cloves (Clov) and Amygdaline (Amyg), is one of the numerous complementary and alternative approaches as an anti-cancer therapy and holds great promise for cancer chemo-prevention with fewer side effects. The current study was designated to assess anti-tumoral immunity and anti-cancer and chemo-preventive effectiveness of herbal extracts of Cur, Ginger, Clov and Amyg in Ehrlich Ascites Carcinoma (EAC)-challenging mice. Chemo-preventive efficacy of herbal extracts of Cur, Gin, Clov and Amyg were analyzed in vivo by examination of the apoptosis rate of EAC tumor cells by flow cytometry. The total numbers of EAC cells, splenocytes counts and leucocytes count with their differentials relative % in peripheral blood (PB) of EACchallenging mice were investigated. EAC-challenging mice treated with herbal extracts of Cur, Gin, Clov and Amyg showed a marked decline in EAC tumor cell count and a noticeable increase in apoptosis rate of EAC tumor cells, a remarkable decrease in serum level of cancer antigen 125 (CA-125) with an obvious increase in the number of splenocytes comparing to that in EAC-challenging mice treated with PBS alone. Moreover, the data indicated an insignificant change in the total leucocytes count and their differentials relative % of eosinophil, neutrophils, monocytes and lymphocytes in EAC-challenging mice treated with Cur and Amyg, but these parameters were markedly increased in EAC-challenging mice injected with Gin and Clov compared to that in EAC-challenging mice treated with PBS alone. To conclude, the herbal extracts of Cur, Gin, Clov and Amyg may have anti-tumoral immunity and anti-cancer potency and potential to reduce the resistance to cancer conventional chemotherapy and exert cancer chemo-protective approaches with low adverse effects. Further research is necessary to determine the regimen's toxicity on various tissues and organs and to connect the diagnostic and therapeutic approaches used in the regimen's biomedical use.

LncRNA MAAMT facilitates macrophage recruitment and proinflammatory activation and exacerbates autoimmune myocarditis through the SRSF1/NF-κB axis

Long non-coding RNAs (lncRNAs) have been implicated in dilated cardiomyopathy (DCM). However, the biological functions and regulatory mechanisms of lncRNAs in DCM remain elusive. Using a mouse model of experimental autoimmune myocarditis (EAM) to mimic DCM, we successfully constructed a dynamic lncRNA expression library for EAM by lncRNA microarray and found that the expression of a macrophage-enriched lncRNA, MAAMT, was significantly increased in the myocardial tissue of mice at the acute stage of EAM. Functionally, MAAMT knockdown alleviated the recruitment and proinflammatory activation of macrophages in the heart, spleen, and peripheral blood of mice at the acute stage of EAM, reduced myocardial inflammation and injury, and eventually reversed ventricular remodelling and improved cardiac function in mice at the chronic stage of EAM. Mechanistically, we identified serine/arginine-rich splicing factor 1 (SRSF1) as an MAAMT-interacting protein in macrophages using RNA pull-down assays coupled with mass spectrometry. MAAMT knockdown attenuated the ubiquitination-mediated degradation of SRSF1, increased the protein expression of SRSF1, and restrained the activation of the NF-κB pathway in macrophages, thereby inhibiting the proinflammatory activation of macrophages. Collectively, our results demonstrate that MAAMT is a key proinflammatory regulator of myocarditis that promotes macrophage activation through the SRSF1-NF-κB axis, providing a new insight into early effective treatment strategies for DCM.

Interspecies regulatory landscapes and elements revealed by novel joint systematic integration of human and mouse blood cell epigenomes.

Knowledge of locations and activities of cis-regulatory elements (CREs) is needed to decipher basic mechanisms of gene regulation and to understand the impact of genetic variants on complex traits. Previous studies identified candidate CREs (cCREs) using epigenetic features in one species, making comparisons difficult between species. In contrast, we conducted an interspecies study defining epigenetic states and identifying cCREs in blood cell types to generate regulatory maps that are comparable between species, using integrative modeling of eight epigenetic features jointly in human and mouse in our Validated Systematic Integration (VISION) Project. The resulting catalogs of cCREs are useful resources for further studies of gene regulation in blood cells, indicated by high overlap with known functional elements and strong enrichment for human genetic variants associated with blood cell phenotypes. The contribution of each epigenetic state in cCREs to gene regulation, inferred from a multivariate regression, was used to estimate epigenetic state regulatory potential (esRP) scores for each cCRE in each cell type, which were used to categorize dynamic changes in cCREs. Groups of cCREs displaying similar patterns of regulatory activity in human and mouse cell types, obtained by joint clustering on esRP scores, harbor distinctive transcription factor binding motifs that are similar between species. An interspecies comparison of cCREs revealed both conserved and species-specific patterns of epigenetic evolution. Finally, we show that comparisons of the epigenetic landscape between species can reveal elements with similar roles in regulation, even in the absence of genomic sequence alignment.

Effect of Sappan Wood Ethanol Extract in CRP Level and Phagocytic Index Between Group of Mice Infected with S. aureus and E. coli

S.aureusand E.coli are pathogenic bacterial that cause many infectious disease in the world. Immunomodulator is needed to prepare the immune system to be able against the infection. Some parameters usually used to assess the immunomodulatory activity such as C-Reactive Protein (CRP) and phagocytic index. This study aims to investigate the difference effect of sappan1wood extract in CRP level and phagocytic index between group mice infected with S. aureus and E. coli. Two treatment groups of mice were prepared for S. aureus and E. coli test. Each group subjected to 7 treatments i.e. (1) Normal mice1(CMC-Na 1 %- Merck® 217277), 1(2) Negative Control (bacterial infection + CMC-Na 1 %),11(3) Positive Control (bacterial infection + Imboost force®1treatment, PT SOHO Industri Pharmasi), (4) Bacterial infection and EESW treatment 25 mg/kg BW, (5) 50 mg/kg BW, (6) 100 mg/kg BW, and (7) 200 mg/kg BW. Mice blood was taken to detect the CRP and phagocytic index after treatment. The T test showed that there was a significant difference between CRP levels (p0.05) and phagocytic index (p0.05) of S. aureus and E. coli group. EESW 200 mg/kg BW reduced CRP level to 11 mg/dL (S. aureus) and 6 mg/dL in (E. coli). EESW 200 mg/kg BW increased phagocytosis to 1.54 folds (S. aureus) and 4.62 folds (E. coli). Sappan wood ethanol extract effect to CRP level and phagocytic index1in mice group infected with E. coli is better than S. aureus1infection group.

Shengqing Jiangzhuo decoction regulates the immunosuppressive microenvironment via the STAT3 signaling pathway for the treatment of malignant ascites in HCC

IntroductionMalignant ascites is a common complication of advanced hepatocellular carcinoma (HCC) that seriously affects the quality of life and survival of patients and has poor clinical efficacy. In the early stages, the authors found that the combination of the Shengqing Jiangzhuo decoction (SQJZD)and Endostar can effectively alleviate the clinical symptoms of malignant ascites in patients with HCC and reduce the amount of malignant ascites. The purpose of this study was to investigate the mechanism of action of SQJZD in HCC-related malignant ascites. MethodsA malignant ascites BALB/c mouse model was established by the intraperitoneal inoculation of H22 cells. The mice were randomly divided into model, Endostar, and high-, medium-, and low-dose traditional Chinese medicine (TCM) combined with Endostar groups. Normal mice from the same batch were used as the normal group. The Endostar and TCM + Endostar groups were intraperitoneally injected with Endostar, whereas the model and normal groups were intraperitoneally injected with equal amounts of normal saline. Different doses of SQJZD were administered to each TCM group, and equal amounts of distilled water were administered to the normal, model, and Endostar groups. ResultsThe malignant ascites volume and body weight were higher in the model group than in the normal group. After treatment, the volume and body weight of mice with malignant ascites decreased, especially in the SQJZD combined with Endostar group. Compared with that in the normal group, the number of regulatory T cells (Tregs) in the peripheral blood and spleen of mice in the model group was significantly decreased; the number of regulatory dendritic cells (DCregs), myeloid-derived suppressor cells (MDSCs), and tumor-associated macrophages (TAMs) in the peripheral blood were not significantly changed; and the number of DCregs, MDSCs, and TAMs in the spleen was increased. Compared with those in the model group, the numbers of Tregs, DCregs, MDSCs, and TAMs in the peripheral blood increased in the Endostar or SQJZD combined with Endostar groups, whereas the numbers of Tregs, DCregs, MDSCs, and TAMs in the spleen decreased. Western blotting revealed that the percentage of p-STAT3-positive cells in the liver and spleen of mice increased in the model group, whereas the percentage of p-STAT3-positive cells in the liver and spleen decreased in mice treated with Endostar or SQJZD combined with Endostar. ConclusionsSQJZD combined with Endostar may attenuate immunosuppression and enhance the antitumor immune response by regulating the STAT3 pathway to alleviate malignant ascites in HCC.

Changes in hematological parameters when using silver preparations in laboratory ICR mice infected with the Newcastle disease virus

The results of blood tests are important indicators for assessing and monitoring the health status, adaptation processes and metabolic states of the body. Currently, there is insufficient data on the effect of the Newcastle disease virus on peripheral blood parameters in animals, and there is also no information on changes in these parameters during Newcastle disease and the simultaneous use of silver preparations in animals. Although the mechanisms of the antiviral action of silver nanoparticles are not yet fully studied, researches show their effectiveness. The purpose of research was to study changes in hematological parameters with using silver preparations in laboratory ICR mice infected with Newcastle disease virus. The object of the study was the blood of laboratory ICR mice, infected with the Newcastle disease virus, and healthy ones as a control, and also silver preparations: Argosil and ArgovitMaxi. As a result of hematological studies, a significant effect of the Newcastle disease virus on hematological parameters was established. Deviations of the main blood parameters in infected animals were demonstrated: there was an increase in the content of granulocytes, the number of erythrocytes, the hemoglobin content and hematocrit; the number of lymphocytes and the average erythrocytes volume were reduced. It was also found that the use of the preparation Argosil significantly reduces the content of monocytes in ICR mice against the background of Newcastle disease viral infection. The use of the preparation Argovit significantly affects the decrease in monocytes and erythrocyte blood parameters: it increases the number of erythrocytes, hemoglobin, hematocrit, and reduces the average hemoglobin content in an erythrocyte, the average hemoglobin concentration in an erythrocyte.

Hemoglobin Hydrolyzate Promotes Iron Absorption in the Small Intestine through Iron Binding Peptides.

Hemoglobin is an excellent source of iron supplements, and its hydrolyzate spontaneously binds iron during digestion and promotes iron absorption in vivo. However, the underlying mechanisms of what peptides bind and how they bind iron ions remain unclear. This study prepared the porcine hemoglobin hydrolyzate through enzymatic hydrolysis and acid treatment and investigated the mechanisms of hemoglobin hydrolyzate on iron absorption through the determination of iron levels in dietary intervention mice, iron binding site analyses, peptide digestion analyses, molecular simulation docking, and INT407 cell validation. The results showed that ingestion of the hemoglobin hydrolyzate diets increased iron levels in the blood of mice, accompanied by the upregulation of duodenal iron circulation-related genes such as ferritin, PCBP1, and HP. Carboxyl, imidazole groups, and aromatic amino acid residues were iron binding sites of hemoglobin hydrolyzate during digestion. VDEVGGEA and VDEVGGE were found to involve the spontaneous and efficient binding of hemoglobin hydrolyzate to iron ions in the intestinal cavity. In particular, the DEVGGE peptide was the typical sequence for hemoglobin hydrolytic peptides to exert iron binding activity.

Single cell transcriptome analysis identified a unique neutrophil type associated with Alzheimer’s disease

BackgroundNeutrophils play an essential role in Alzheimer’s disease (AD) pathology. However, the extent of their heterogeneity remains poorly explored, particularly in the context of developing novel therapies targeting these cells.ResultsWe investigate the population structure of neutrophils purified from peripheral blood samples of AD mice. Utilizing single cell RNA sequencing, we comprehensively map neutrophil populations into six distinct clusters and find that the Neu-5 subset is specially enriched in AD mice. This subset exhibits fewer specific granules and a lower mature score. Gene ontology (GO) analysis reveals that genes involved in cytokine-mediated signaling are downregulated in the Neu-5 cluster. Furthermore, we identify the Ccrl2 gene is specifically upregulated in this subgroup, which is confirmed by flow cytometry in AD mice. Finally, immunohistochemical staining indicates that CCRL2 protein is increased in the brains of AD mice.ConclusionsWe identify a unique CCRL2 positive neutrophil cluster, that is specifically enriched in the peripheral blood of AD mice.

Cystathionine γ-lyase-derived H2S negatively regulates thymic egress via allosteric inhibition of sphingosine-1-phosphate lyase.

Thymic egress is a crucial process for thymocyte maturation, strictly regulated by sphingosine-1-phosphate lyase (S1PL). Recently, cystathionine γ-lyase (CSE), one of the enzymes producing hydrogen sulfide (H2S), has emerged as a vital immune process regulator. However, the molecular connection between CSE, H2S and thymic egress remains largely unexplored. In this study, we investigated the regulatory function of CSE in the thymic egress of immune cells. We showed that genetic knockout of CSE or pharmacological inhibition by CSE enzyme inhibitor NSC4056 or D,L-propargylglycine (PAG) significantly enhanced the migration of mature lymphocytes and monocytes from the thymus to the peripheral blood, and this redistribution effect could be reversed by treatment with NaHS, an exogenous donor of H2S. In addition, the CSE-generated H2S significantly increased the levels of S1P in the peripheral blood, thymus and spleen of mice, suppressed the production of proinflammatory cytokines and rescued pathogen-induced sepsis in cells and in vivo. Notably, H2S or polysulfide inhibited S1PL activity in cells and an in vitro purified enzyme assay. We found that this inhibition relied on a newly identified C203XC205 redox motif adjacent to the enzyme's active site, shedding light on the biochemical mechanism of S1PL regulation. In conclusion, this study uncovers a new function and mechanism for CSE-derived H2S in thymic egress and provides a potential drug target for treating S1P-related immune diseases.

CDK4/6 inhibition enhances T-cell immunotherapy on hepatocellular carcinoma cells by rejuvenating immunogenicity

Hepatocellular carcinoma (HCC) poses a significant clinical challenge, necessitating the integration of immunotherapeutic approaches. Palbociclib, a selective CDK4/6 inhibitor, has demonstrated promising efficacy in preclinical HCC models and is being evaluated as a novel therapeutic option in clinical trials. Additionally, CDK4/6 inhibition induces cellular senescence, potentially influencing the tumor microenvironment and immunogenicity of cancer cells. In this study, we conducted comprehensive bioinformatic analyses using diverse HCC transcriptome datasets, including bulk and single-cell RNA-sequencing data from public databases. We also utilized human and mouse HCC cells to investigate functional aspects. Primary T cells isolated from mouse blood were employed to assess T cell immunity against HCC cells. Results revealed that CD8+ T-cell infiltration correlates with improved outcomes in HCC patients with suppressed CDK4/6 expression. Moreover, CDK4/6 expression was associated with alterations in the immune landscape and immune checkpoint expression within the liver tumor microenvironment. Furthermore, we found that treatment with Palbociclib and Doxorubicin induces cellular senescence and a senescence-associated secretory phenotype in HCC cells. Notably, pretreatment with Palbociclib augmented T cell-mediated cytotoxicity against HCC cells, despite upregulation of PD-L1, surpassing the effects of Doxorubicin pretreatment. In conclusion, our study elucidates a novel mechanism by which CDK4/6 inhibition enhances T-cell-associated cancer elimination and proposes a potential therapeutic strategy to enhance T-cell immunotherapy on HCC.Graphical abstract

The Alzheimer's disease-linked protease BACE2 cleaves VEGFR3 and modulates its signaling.

The β-secretase β-site APP cleaving enzyme (BACE1) is a central drug target for Alzheimer's disease. Clinically tested, BACE1-directed inhibitors also block the homologous protease BACE2. Yet little is known about physiological BACE2 substrates and functions in vivo. Here, we identify BACE2 as the protease shedding the lymphangiogenic vascular endothelial growth factor receptor 3 (VEGFR3). Inactivation of BACE2, but not BACE1, inhibited shedding of VEGFR3 from primary human lymphatic endothelial cells (LECs) and reduced release of the shed, soluble VEGFR3 (sVEGFR3) ectodomain into the blood of mice, nonhuman primates, and humans. Functionally, BACE2 inactivation increased full-length VEGFR3 and enhanced VEGFR3 signaling in LECs and also in vivo in zebrafish, where enhanced migration of LECs was observed. Thus, this study identifies BACE2 as a modulator of lymphangiogenic VEGFR3 signaling and demonstrates the utility of sVEGFR3 as a pharmacodynamic plasma marker for BACE2 activity in vivo, a prerequisite for developing BACE1-selective inhibitors for safer prevention of Alzheimer's disease.

Expression characteristics of lipid metabolism-related genes and correlative immune infiltration landscape in acute myocardial infarction

Lipid metabolism is an important part of the heart's energy supply. The expression pattern and molecular mechanism of lipid metabolism-related genes (LMRGs) in acute myocardial infarction (AMI) are still unclear, and the link between lipid metabolism and immunity is far from being elucidated. In this study, 23 Common differentially expressed LMRGs were discovered in the AMI-related mRNA microarray datasets GSE61144 and GSE60993. These genes were mainly related to “leukotriene production involved in inflammatory response”, “lipoxygenase pathway”, “metabolic pathways”, and “regulation of lipolysis in adipocytes” pathways. 12 LMRGs (ACSL1, ADCY4, ALOX5, ALOX5AP, CCL5, CEBPB, CEBPD, CREB5, GAB2, PISD, RARRES3, and ZNF467) were significantly differentially expressed in the validation dataset GSE62646 with their AUC > 0.7 except for ALOX5AP (AUC = 0.699). Immune infiltration analysis and Pearson correlation analysis explored the immune characteristics of AMI, as well as the relationship between these identified LMRGs and immune response. Lastly, the up-regulation of ACSL1, ALOX5AP, CEBPB, and GAB2 was confirmed in the mouse AMI model. Taken together, LMRGs ACSL1, ALOX5AP, CEBPB, and GAB2 are significantly upregulated in AMI patients' blood, peripheral blood of AMI mice, myocardial tissue of AMI mice, and therefore might be new potential biomarkers for AMI.

Preventive effects of Chinese Xinjiang naturally fermented yogurt separated from Lactobacillus rhamnosusAFY02 on acute gouty arthritis in mice

AbstractLactobacillus rhamnosus AFY02 (LR‐AFY02) is a newly discovered strain isolated and identified from naturally fermented yogurt in Xinjiang, China. This research aims to investigate the mechanism of action of LR‐AFY02 in mice with acute gouty arthritis. We examined the degree of foot swelling, pain threshold, blood biochemical indicators, histopathological changes, and mRNA expression. LR‐AFY02 can decrease the severity of mouse foot edema and raise the pain threshold. LR‐AFY02 can increase the enzyme activity of superoxide dismutase (SOD) and the level of glutathione (GSH) while lowering the enzyme activity of myeloperoxidase (MPO) and the level of malondialdehyde (MDA) in mice with acute arthritis. Interleukin‐6 (IL‐6), IL‐10, interleukin‐1 beta (IL‐1β), and tumor necrosis factor‐alpha (TNF‐α) levels in the blood of mice with acute arthritis are also decreased by LR‐AFY02. Histopathological findings demonstrated that LR‐AFY02 reduced tissue damage in the mouse foot and ankle joints. LR‐AFY02 may suppress the mRNA expression of extracellular signal‐regulated kinase 1/2 (ERK1/2), cyclooxygenase‐2 (COX‐2), prostaglandin E2 (PGE2), IL‐6, interferon gamma (IFN‐γ), and TNF‐α in the tissues of the ankle joints of mice with acute arthritis. Additionally, LR‐AFY02 has the ability to increase the expression of catalase (CAT), manganese superoxide dismutase (Mn‐SOD), and copper/zinc superoxide dismutase (Cu/Zn‐SOD). As a result, it is clear that L. rhamnosus AFY02 is more effective than glucosamine sulfate at preventing acute gouty arthritis.

Preparation, Physicochemical, and Cyto- and Genotoxic Characterisation of Polysaccharide Composites Containing Carbon Quantum Dots.

Rapid industrial growth is associated with an increase in the production of environmentally harmful waste. A potential solution to significantly reduce pollution is to replace current synthetic materials with readily biodegradable plastics. Moreover, to meet the demands of technological advancements, it is essential to develop materials with unprecedented properties to enhance their functionality. Polysaccharide composites demonstrate significant potential in this regard. Polysaccharides possess exceptional film-forming abilities and are safe for human use, biodegradable, widely available, and easily modifiable. Unfortunately, polysaccharide-based films fall short of meeting all expectations. To address this issue, the current study focused on incorporating carbon quantum dots (CQDs), which are approximately 10 nm in size, into the structure of a starch/chitosan biocomposite at varying concentrations. This modification has improved the mechanical properties of the resulting nanocomposites. The inclusion of nanoparticles led to a slight reduction in solubility and an increase in the swelling degree. The optical characteristics of the obtained films were influenced by the presence of CQDs, and the fluorescence intensity of the nanocomposites changed due to the specific heavy metal ions and amino acids used. Consequently, these nanocomposites show great potential for detecting these compounds. Cellular viability assessments and comet assays confirm that the resulting nanocomposites do not exhibit any cytotoxic properties based on this specific analytic method. The tested nanocomposites with the addition of carbon quantum dots (NC/CD II and NC/CD III) were characterised by greater genotoxicity compared to the negative control. The positive control, the starch/chitosan composite alone, was also characterised by a greater induction of chromatin damage in mouse cells compared to a pure mouse blood sample.

Dissecting Acute Drug-Induced Hepatotoxicity and Therapeutic Responses of Steatotic Liver Disease Using Primary Mouse Liver and Blood Cells in a Liver-On-A-Chip Model.

Metabolic dysfunction-associated steatotic liver disease (MASLD) is hallmarked by hepatic steatosis, cell injury, inflammation, and fibrosis. This study elaborates on a multicellular biochip-based liver sinusoid model to mimic MASLD pathomechanisms and investigate the therapeutic effects of drug candidates lanifibranor and resmetirom. Mouse liver primary hepatocytes, hepatic stellate cells, Kupffer cells, and endothelial cells are seeded in a dual-chamber biocompatible liver-on-a-chip (LoC). The LoC is then perfused with circulating immune cells (CICs). Acetaminophen (APAP) and free fatty acids (FFAs) treatment recapitulate acute drug-induced liver injury and MASLD, respectively. As a benchmark for the LoC, multiplex immunofluorescence on livers from APAP-injected and dietary MASLD-induced mice reveals characteristic changes on parenchymal and immune cell populations. APAP exposure induces cell death in the LoC, and increased inflammatory cytokine levels in the circulating perfusate. Under FFA stimulation, lipid accumulation, cellular damage, inflammatory secretome, and fibrogenesis are increased in the LoC, reflecting MASLD. Both injury conditions potentiate CIC migration from the perfusate to the LoC cellular layers. Lanifibranor prevents the onset of inflammation, while resmetirom decreases lipid accumulation in hepatocytes and increases the generation of FFA metabolites in the LoC. This study demonstrates the LoC potential for functional and molecular evaluation of liver disease drug candidates.

12/15-Lipooxygenase Inhibition Reduces Microvessel Constriction and Microthrombi after Subarachnoid Hemorrhage in Mice.

Impaired cerebral circulation, induced by blood vessel constrictions and microthrombi, leads to delayed cerebral ischemia after subarachnoid hemorrhage (SAH). 12/15-Lipooxygenase (12/15-LOX) overexpression has been implicated in worsening early brain injury outcomes following SAH. However, it is unknown if 12/15-LOX is important in delayed pathophysiological events after SAH. Since 12/15-LOX produces metabolites that induce inflammation and vasoconstriction, we hypothesized that 12/15-LOX leads to microvessel constriction and microthrombi formation after SAH, and thus 12/15-LOX is an important target to prevent delayed cerebral ischemia. SAH was induced in C57BL/6 and 12/15-LOX-/- mice of both sexes by endovascular perforation. Expression of 12/15-LOX was assessed in brain tissue slices and in vitro. C57BL/6 mice were administered either ML351 (12/15-LOX inhibitor) or vehicle. Mice were evaluated for daily neuroscore and euthanized on day five to assess cerebral 12/15-LOX expression, vessel constrictions, platelet activation, microthrombi, neurodegeneration, infarction, cortical perfusion, and for development of delayed deficits. Finally, the effect of 12/15-LOX inhibition on platelet activation was assessed in SAH patient samples using a platelet spreading assay. In SAH mice, 12/15-LOX was upregulated in brain vascular cells and there was an increase in 12-S-HETE. Inhibition of 12/15-LOX improved brain perfusion on days 4-5 and attenuated delayed pathophysiological events, including microvessel constrictions, microthrombi, neuronal degeneration, and infarction. Additionally, 12/15-LOX inhibition reduced platelet activation in human and mouse blood samples. Cerebrovascular 12/15-LOX overexpression plays a major role in brain dysfunction after SAH by triggering microvessel constrictions and microthrombi formation, which reduces brain perfusion. Inhibiting 12/15-LOX may be a therapeutic target to improve outcomes after SAH.

Autoantibodies-Abzymes with Amylase Activity in Experimental Autoimmune Encephalomyelitis Mice

The exact mechanisms of multiple sclerosis evolution are still unknown. However, the progress in C57BL/6 mice of experimental autoimmune encephalomyelitis (EAE, similar to human multiple sclerosis) happens as a result of the contravention of bone marrow hematopoietic stem cells differentiation profiles integrated with the production of toxic for human’s auto-antibodies-abzymes splitting myelin basic protein (MBP), myelin oligodendrocyte glycoprotein (MOG), histones, DNA and RNA as well as IgGs with catalase and peroxidase activities. Here, we first analyzed the change in the relative amylase activity of IgGs from C57BL/6 mice blood over time at different stages of the EAE evolution: onset, acute, and remission phases. It was shown that the amylase activity of IgGs during spontaneous development of EAE first noticeably increases from 50 to 80, then sharply decreases to 100-130, and again sharply increases to 150 days of mice life. Immunization of 3-month-old mice (time zero) with the DNA-histone complex leads to a decrease, while their treatment with MOG results in a substantial increase in the amylase activity of IgGs. The relative activities of all IgG-abzymes indicated above corresponding to various stages of EAE development were compared. The data shows that spontaneous EAE and immunization of mice with DNAhistones complex and MOG leads to a different and very expanded formation of various B lymphocytes, producing abzymes with several diverse catalytic activities. In addition, the relative number of lymphocytes producing diverse antibodies with distinct enzymatic activities (including their relative activities) significantly depend both on the stages of spontaneous and antigen-induced accelerations of EAE development.

Ozone enhances the efficacy of radiation therapy in esophageal cancer.

Radioresistance is increasingly developed in esophageal cancer. Increasing radiation sensitivity can reduce the mortality of esophageal cancer. To investigate the effect and mechanism of ozone on the radiotherapy sensitization of esophageal carcinoma. KYSE150 cells were xenografted subcutaneously into nude mice and irradiated with 8Gy radiation according to different subgroups (sham, radiation, ozone and radiation+ozone group (n = 10 per group)). Half of the mice were used to determine the body weight, tumor size and tumor weight. Half of the mice were used to collect peripheral blood. The serum was centrifuged to detect circulating cell-free DNA (cf-DNA), interleukin-6 (IL-6), interferon-γ (IFN-γ), myeloperoxidase (MPO)-DNA complexes, tumor necrosis factor-α (TNF-α), matrix metalloproteinase-9 (MMP-9) and hypoxia-inducible factor-1α (HIF-1α) using commercial kits. The levels of phosphorylation AMP-activated protein kinase (p-AMPK) and scavenger receptor-A (SR-A) were measured by immunocytochemistry and Western blotting in the tumor tissues of mice. Ozone alone or combined with radiation therapy significantly reduced the body weight, tumor volume and tumor weight of esophageal cancer compared to the sham group. The ELISA results showed that the levels of cf-DNA, IFN-γ, MPO-DNA complexes, TNF-α, IL-6, HIF-1α and MMP-9 in the peripheral blood of mice treated with ozone combined with radiation were significantly lower compared with the radiation group. Ozone, synergistically with radiation, significantly increased the protein expression of p-AMPK and SR-A. Ozone may increase the radiosensitivity of esophageal cancer by inhibiting neutrophil extracellular traps.

Clinical features associated with the presence of anti-Ro52 and anti-Ro60 antibodies in Jo-1 antibody-positive anti-synthetase syndrome.

Anti-SSA antibodies target two unrelated proteins, Ro52 (E3 ligase) and Ro60 (RNA binding protein). Previous studies indicate that anti-Ro52 antibodies are frequently associated with various myositis-specific autoantibodies (MSAs)-including anti-tRNA synthetase antibodies-and that the coexistence of MSAs and anti-Ro52 antibodies may portend worse clinical outcomes. Although not well-described in the setting of myositis, work from our animal model of HRS (histidyl-tRNA synthetase)-induced myositis suggests that anti-Ro60 antibodies may also be linked to specific MSAs such as anti-HRS/Jo-1. We therefore aimed to demonstrate the prevalence and clinical characteristics of Ro52 and Ro60 antibody positivity in patients possessing Jo-1 antibodies. To establish the immunological link between anti-synthetase, anti-Ro52, and anti-Ro60 antibodies, we evaluated the relative titers of these antibodies in blood and bronchoalveolar lavage fluid (BALF) of mice following immunization with HRS/Jo-1. In parallel, we used ELISA-based approaches to assess sera from 177 anti-Jo1 antibody-positive patients for the presence of anti-Ro52 and/or anti-Ro60 antibodies. We then determined statistical associations between co-existing anti-Jo-1, anti-Ro52, and/or anti-Ro60 antibodies and clinical manifestations associated with the anti-synthetase syndrome. Mice immunized with HRS had higher levels of anti-Ro52 and anti-Ro60 antibodies in serum and BALF than PBS-immunized mice. In 177 anti-Jo-1 antibody-positive patients, the prevalence of anti-Ro52 and anti-Ro60 antibodies was 36% and 15%, respectively. The frequency of dry eye/dry mouth, interstitial pneumonia, and pulmonary events over time differed between patients with various combinations of anti-Ro52 and anti-Ro60 antibodies. While anti-Ro52 antibodies generally correlated with statistically significant increases in each of these clinical manifestations, the presence of Ro60 antibodies alone was associated with decreased frequency of ILD. Anti-Ro52 and/or anti-Ro60 antibodies are often co-expressed with anti-Jo1 antibodies, defining clinical subsets with different disease course/outcomes.

Establishment of a humanized ST6GAL1 mouse model for influenza research.

This study aimed to construct and characterize a humanized influenza mouse model expressing hST6GAL1. Humanized fragments, consisting of the endothelial cell-specific K18 promoter, human ST6GAL1-encoding gene, and luciferase gene, were microinjected into the fertilized eggs of mice. The manipulated embryos were transferred into the oviducts of pseudopregnant female mice. The offspring were identified using PCR. Mice exhibiting elevated expression of the hST6GAL1 gene were selectively bred for propagation, and invivo analysis was performed for screening. Expression of the humanized gene was tested by performing immunohistochemical (IHC) analysis. Hematologic and biochemical analyses using the whole blood and serum of humanized hST6GAL1 mice were performed. Successful integration of the human ST6GAL1 gene into the mouse genome led to the overexpression of human SiaT ST6GAL1. Seven mice were identified as carrying copies of the humanized gene, and the invivo analysis indicated that hST6GAL1 gene expression in positive mice mirrored influenza virus infection characteristics. The IHC results revealed that hST6GAL1 was expressed in the lungs of humanized mice. Moreover, the hematologic and biochemical parameters of the positive mice were within the normal range. A humanized influenza mouse model expressing the hST6GAL1 gene was successfully established and characterized.