HuR coordinates systemic aging through platelet infiltration.

C-X-C motif chemokine receptor 3 (CXCR3) is an important receptor controlling the migration of leukocytes, although there is no report regarding its role in Neospora caninum infection. Herein, we investigated the relevance of CXCR3 in the resistance mechanism to N. caninum infection in mice. Wild-type (WT) C57BL/6 mice and CXCR3-knockout (CXCR3KO) mice were used in all experiments. WT mice displayed a high survival rate (100%), while 80% of CXCR3KO mice succumbed to N. caninum infection within 50 days. Compared with WT mice, CXCR3KO mice exhibited significantly lower body weights and higher clinical scores at the subacute stage of infection. Flow cytometric analysis revealed CXCR3KO mice as having significantly increased proportions and numbers of CD11c-positive cells compared with WT mice at 5 days post infection (dpi). However, levels of interleukin-6 and interferon-γ in serum and ascites were similar in all groups at 5 dpi. Furthermore, no differences in parasite load were detected in brain, spleen, lungs or liver tissue of CXCR3KO and WT mice at 5 and 21 dpi. mRNA analysis of brain tissue collected from infected mice at 30 dpi revealed no changes in expression levels of inflammatory response genes. Nevertheless, the brain tissue of infected CXCR3KO mice displayed significant necrosis and microglial activation compared with that of WT mice at 21 dpi. Interestingly, the brain tissue of CXCR3KO mice displayed significantly lower numbers of FoxP3+ cells compared with the brain tissue of WT mice at 30 dpi. Accordingly, our study suggests that the lack of active regulatory T cells in brain tissue of infected CXCR3KO mice is the main cause of these mice having severe necrosis and lower survival compared with WT mice. Thus, CXCR3+ regulatory T cells may play a crucial role in control of neosporosis.

To investigate anti-aging effects of probiotic-fermented kelp enzymatic hydrolysate culture (KMF), probiotic-fermented kelp enzymatic hydrolysate supernatant (KMFS), and probiotic-fermented kelp enzymatic hydrolysate bacteria suspension (KMFP) in D-galactose-induced aging mice. The study uses a probiotic-mixture of Lactobacillus reuteri, Pediococcus pentosaceus, and Lactobacillus acidophilus strains for kelp fermentation. KMF, KMFS, and KMFP prevent D-galactose-induced elevation of malondialdehyde levels in serum and brain tissue of aging mice, and they increase superoxide dismutase and catalase levels and total antioxidant capacity. Furthermore, they improve the cell structure of mouse brain, liver, and intestinal tissue. Compared with the model control group, the KMF, KMFS, and KMFP treatments regulate mRNA and protein levels of genes associated with aging, the concentrations of acetic acid, propionic acid, and butyric acid in the three treatment groups are more than 1.4-, 1.3-, and 1.2-fold increased, respectively. Furthermore, the treatments affect the gut microbiota community structures. These results suggest that KMF, KMFS, and KMFP can modulate gut microbiota imbalances and positively affect aging-related genes to achieve anti-aging effects.

Evaluation of immunogenicity and protective efficacy of a novel Senecavirus A strain-based inactivated vaccine in mice

Heme chloride (Hemin) is an in vitro purified form of natural heme and an important raw material for anti-anemia and antitumor drugs. This study aims to analyze the protective effect of Hemin on tissue damage in low-pressure oxygen chamber simulated plateau hypoxic mice, and explore its role in anti-plateau hypoxia. Thirty male BALB/c mice were randomly divided into a blank group, a positive drug group (acetazolomide, 200 mg/kg), a Hemin low-dose group (15 mg/kg), a Hemin medium-dose group (30 mg/kg), and a Hemin high-dose group (60 mg/kg) with intraperitoneal injection. The anti-hypoxic activity of Hemin was explored by atmospheric closed hypoxia experiment and the optimal dose was screened. Thirty-six male BALB/c mice were randomly divided into a blank group, a hypoxia group, a positive drug group, and a Hemin high-dose group. The plasma inflammatory factor levels and oxidative stress indicators malondialdehyde (MDA), glutataione (GSH), and superoxide dismutase (SOD) levels of myocardium, brain, lung, and liver tissues were measured in different groups with hypoxia for 24 h. The degree of histopathological damage of mice was observed with HE staining. The degree of protection of Hemin against tissue hypoxia injury was detected with the hypoxia probe piperidazole. Compared with the blank group, the survival time of mice in the positive drug group, the Hemin medium-dose group, and high-dose group was significantly extended (all P<0.05), with the highest prolongation rate in the Hemin high-dose group. Compared with the hypoxia group, mice in the Hemin high-dose group showed a significant increase in SOD level and GSH content of brain tissue, and a significant decrease in MDA content of lung tissue (all P<0.05). The results of HE staining and hypoxia probe showed that Hemin had a significant protective effect on the damage of liver, heart, brain and lung tissues of mice with hypoxia, and the most obvious effect on that of the brain tissue. Hemin has an effect of improvement on oxidative stress and inflammatory response caused by hypoxia, and has obvious protective effect on tissue damage caused by hypoxia.

To assess the industrial possibility of mixed-extracts containing Hovenia dulcis Thunberg and 12 different botanical ingredients, a protective effect was confirmed in the chronic ethanol-induced the liver, brain, and blood injury in mouse. Blood glucose levels of the normal control group(NG) and ethanol administration group(EG) were respectively 119.43mg/dL and 305.25mg/dL, and the mixed-extracts administration group(100, 200mg/kg body weight + 25% ethanol 5g/kg body weight respectively; ME100 & ME200) were decreased to 272.76mg/dL and Corresponding author: Ho Jin Heo Tel: +82-55-772-1907 Fax: +82-55-772-1909 E-mail: hjher@gnu.ac.kr 126 ... Journal of Agriculture & Life Science 50(2) 234.60mg/dL. Blood ethanol contents were decreased in ME100 and ME200(3.85mg/dL, 3.08mg/dL) compared to EG(4.08mg/dL), and blood acetaldehyde contents were also decreased in ME(15.76mg/dL, 15.16mg/dL) compared to EG(18.72mg/dL). The contents of hepatotoxic indicators such as glutamine pyruvic transaminase(GPT) and glutamic oxaloacetic transaminase (GOT), nephrotoxic indicators such as blood urea nitrogen(BUN), and creatine(CRE), and total cholestero(TCHO), and triglyceride(TG) in mouse blood serum were significantly decreased in the ME compared to EG. The acetylcholinesterase(AChE) activity of ME(109.00% and 108.47%, respectively) in mouse brain tissues was decreased in ME compared to EG(116.10%). Finally, ME was remarkable in vivo antioxidant activities in the mouse liver and brain tissues by superoxide dismutase(SOD), oxidized glutathione(GSH)/total GSH ratio and the malondialdehyde (MDA) assay. Therefore, the mixed-extracts was considered to be effective a high value food with protective effect against chronic ethanol traetment-induced cytotoxicity in liver and brain tissues.

Learning and memory disorders arise from distinct age-associated processes, and aging animals are often used as a model of memory impairment. The root of Polygala tenuifolia has been commonly used in some Asian countries as memory enhancer and its memory improvement has been reported in various animal models. However, there is less research to verify its effect on memory functions in aged animals. Herein, the memory-enhancing effects of the crude extract of Polygala tenuifolia (EPT) on normal aged mice were assessed by Morris water maze (MWM) and step-down passive avoidance tests. In MWM tests, the impaired spatial memory of the aged mice was partly reversed by EPT (100 and 200 mg/kg; P < 0.05) as compared with the aged control mice. In step-down tests, the nonspatial memory of the aged mice was improved by EPT (100 and 200 mg/kg; P < 0.05). Additionally, EPT could increase superoxide dismutase (SOD) and catalase (CAT) activities, inhibit monoamine oxidase (MAO) and acetyl cholinesterase (AChE) activities, and decrease the levels of malondialdehyde (MDA) in the brain tissue of the aged mice. The results showed that EPT improved memory functions of the aged mice probably via its antioxidant properties and via decreasing the activities of MAO and AChE.

Objective: The mechanism of cerebral ischemia-reperfusion injury is closely related to inflammation, and miRNA-135b-5p may play a key role in the inflammation of cerebral ischemia injury. In this study we investigate the effect of miRNA-135b-5p through GSK3 β-TLR3 signal axis on inflammatory response in mice with cerebral ischemia-reperfusion injury. Methods: 48 mice were randomly divided into sham operation group (n = 12), model group (n = 12), negative control group (n = 12) and overexpression group (n = 12). After neurobehavioral score, the left brain of mice was killed and the water content of brain tissue was measured, the expression levels of TNF- α and IL-6 in brain tissue and serum were detected by ELISA method, the expression levels of TLR3, GSK3β and ERK protein in brain tissue were detected by Western Blot, and the mRNA expression levels of TLR3, GSK3 β and ERK in each group were detected by RT-PCR. Results: The neurobehavioral scores in the model group, negative control group and overexpression group were significantly higher than those in the sham operation group (P < 0.05), and the neurobehavioral scores in the overexpression group were significantly lower than those in the negative control group and model group (P < 0.05). The expression level of miRNA-135b-5p in brain tissue of mice in model group and negative control group was significantly lower than that in sham operation group, while the expression level of miRNA-135b-5p in overexpression group was significantly higher than that in other three groups. The expression levels of TNF- α and IL-6 in brain tissue and serum of mice in model group and negative control group were significantly higher than those in sham operation group, while the expression levels of TNF- α and IL-6 in overexpression group were significantly lower than those in model group and negative control group. The mRNA expression levels of TLR3, GSK3 β and ERK in the model group and negative control group were significantly higher than those in the sham operation group, while the mRNA expression levels of TLR3, GSK3 β and ERK in the overexpression group were significantly lower than those in the model group and negative control group. Conclusion: Overexpression of miRNA-135b-5p could inhibit the expression of GSK3β-TLR3 signal axis related genes and reduce the inflammatory response during cerebral ischemia-reperfusion injury, which may be a new strategy for the treatment of cerebral ischemia-reperfusion injury.

In cancers such as malignant melanoma, tumor spread or metastasis occurs preferentially to certain organ sites. Mouse B16 melanoma cell sublines that have been selected sequentially in vivo for enhanced blood-borne colonization of lung, brain, or ovary were tested for their survival and growth stimulation in vitro by soluble factors released from suspensions of mouse lung, brain, liver, ovary or kidney tissues. In general, the growth rate of lung-colonizing B16 cells was stimulated by high concentrations of lung-tissue-derived factors significantly more than by factors from the other tissues, whereas the growth rate of ovary-colonizing B16 cells was stimulated by ovary or lung-tissue-derived factors significantly more than by factors from the other tissues. In contrast, the growth of brain-colonizing B16 cells was not stimulated by factors released from brain tissue. When it occurred, stimulation of B16 cell growth by factors released from mouse organ tissues was dose-dependent. Liver tissue factors, and at high concentrations kidney tissue factors, inhibited cell growth of the B16 sublines, an inhibition correlating with the low potential of B16 cells to colonize liver and kidney in vivo. In addition to preferential target-organ cell adhesion found previously with B16 sublines (Nicolson et al., 1985a), the present results suggest that metastasis to specific organ sites is also dependent on the survival and growth of B16 cells affected by soluble organ-derived factors.

To investigate the development and dynamic changes of cysts in the brain of mice following infection with different forms of Toxoplasma gondii, so as to provide insights into for toxoplasmosis prevention and control. ICR mice at ages of 6 to 8 weeks, each weighing 20 to 25 g, were intraperitoneally injected with tachyzoites of the T. gondii PRU strain at a dose of 1 × 105 tachyzoites per mouse, orally administered with cysts at a dose of 20 oocysts per mouse or oocysts at a dose of 200 oocysts per mouse for modeling chronic T. gondii infection in mice, and the clinical symptoms and survival of mice were observed post-infection. Mice were orally infected with T. gondii cysts at doses of 10 (low-dose group), 20 (medium-dose group), 40 cysts per mouse (high-dose group), and the effect of different doses of T. gondii infections on the number of cysts was examined in the mouse brain. Mice were orally administered with T. gondii cysts at a dose of 20 cysts per mouse, and grouped according to gender (female and male) and time points of infections (20, 30, 60, 90, 120, 150, 180 days post-infection), and the effects of gender and time points of infections on the number of cysts was examined in the mouse brain. In addition, mice were divided into the tachyzoite group (Group T), the first-generation cyst group (Group C1), the second-generation cyst group (Group C2), the third-generation cyst (Group C3) and the fourth-generation cyst group (Group C4). Mice in the Group T were intraperitoneally injected with T. gondii tachyzoites at a dose of 1 × 105 tachyzoites per mouse, and the cysts were collected from the mouse brain tissues 30 days post-infection, while mice in the Group C1 were orally infected with the collected cysts at a dose of 30 cysts per mouse. Continuous passage was performed by oral administration with cysts produced by the previous generation in mice, and the effect of continuous passage on the number of cysts was examined in the mouse brain. Following infection with T. gondii tachyzoites, cysts and oocysts in mice, obvious clinical symptoms were observed on days 6 to 13 and mice frequently died on days 7 to 12. The survival rates of mice were 67.0%, 87.0% and 53.0%, and the mean numbers of cysts were (516.0 ± 257.2), (1 203.0 ± 502.0) and (581.0 ± 183.1) in the mouse brain (F = 11.94, P < 0.01) on day 30 post-infection with T. gondii tachyzoites, cysts and oocysts, respectively, and the numbers of cysts in the brain tissues were significantly lower in mice infected with T. gondii tachyzoites and oocysts than in those infected with cysts (all P values < 0.01). The survival rates of mice were 87.0%, 87.0% and 60.0%, and the mean numbers of cysts were (953.0 ± 355.5), (1 084.0 ± 474.3) and (1 113.0 ± 546.0) in the mouse brain in the low-, medium- and high-dose groups on day 30 post-infection, respectively (F = 0.42, P > 0.05). The survival rates of male and female mice were 73.0% and 80.0%, and the mean numbers of cysts were (946.4 ± 411.4) and (932.1 ± 322.4) in the brain tissues of male and female mice, respectively (F = 1.63, P > 0.05). Following continuous passage, the mean numbers of cysts were (516.0 ± 257.2), (1 203.0 ± 502.0), (896.8 ± 332.3), (782.5 ± 423.9) and (829.2 ± 306.0) in the brain tissues of mice in the T, C1, C2, C3 and C4 groups, respectively (F = 4.82, P < 0.01), and the number of cysts was higher in the mouse brain in Group 1 than in Group T (P < 0.01). Following oral administration of 20 T. gondii cysts in mice, cysts were found in the moues brain for the first time on day 20 post-infection, and the number of cysts gradually increased over time, peaked on days 30 and 90 post-infection and then gradually decreased; however, the cysts were still found in the mouse brain on day 180 post-infection. There is a higher possibility of developing chronic T. gondii infection in mice following infection with cysts than with oocysts or tachyzoites and the most severe chronic infection is seen following infection with cysts. The number of cysts does not correlate with the severity of chronic T. gondii infection, and the number of cysts peaks in the mouse brain on days 30 and 90 post-infection.

Age, calorie restriction, and age of calorie restriction onset reduce maturation of natural killer cells in C57Bl/6 mice

Cathepsin C (CC) (EC 3.4.14.1, dipeptidyl peptidase I) is a lysosomal cysteine protease that is required for the activation of several granule-associated serine proteases in vivo. CC has been shown to be constitutively expressed in various tissues, but the enzyme is hardly detectable in central nervous system (CNS) tissues. In the present study, we investigated the regional and cellular distribution of CC in normal, aging and pathological mouse brains. Immunoblotting failed to detect CC protein in whole brain tissues of normal mice, as previously described. However, low proteolytic activity of CC was detected in a brain region-dependent manner, and granular immunohistochemical signals were found in neuronal perikarya of particular brain regions, including the accessory olfactory bulb, the septum, CA2 of the hippocampus, a part of the cerebral cortex, the medial geniculate, and the inferior colliculus. In aged mice, the number of CC-positive neurons increased to some extent. The protein level of CC and its proteolytic activity showed significant increases in particular brain regions of mouse models with pathological conditions--the thalamus in cathepsin D-deficient mice, the hippocampus of ipsilateral brain hemispheres after hypoxic-ischemic brain injury, and peri-damaged portions of brains after penetrating injury. In such pathological conditions, the majority of the cells that were strongly immunopositive for CC were activated microglia. These lines of evidence suggest that CC is involved in normal neuronal function in certain brain regions, and also participates in inflammatory processes accompanying pathogenesis in the CNS.

Immunomodulatory effect of pachymaran on cyclosporine A (CsA)-induced lung injury in mice

Vascular endothelial growth factor (VEGF) serves an important role in promoting angiogenesis and tissue regeneration. However, the lack of an effective delivery system that can target this growth factor to the injured site reduces its therapeutic efficacy. Therefore, in the current study, collagen-binding VEGF was constructed by fusing a collagen-binding domain (CBD) to the N-terminal of native VEGF. The CBD-VEGF can specifically bind to collagen which is the major component of the extracellular matrix in fibrotic liver. The anti-fibrotic effects of this novel material were investigated by the carbon tetrachloride (CCl4)-induced liver fibrotic mouse model. Mice were injected with CCl4 intraperitoneally to induce liver fibrosis. CBD-VEGF was injected directly into the liver tissue of mice. The liver tissues were stained with hematoxylin and eosin for general observation or with Masson's trichrome staining for detection of collagen deposition. The hepatic stellate cell activation, blood vessel formation and hepatocyte proliferation were measured by immunohistochemical staining for α-smooth muscle actin, CD31 and Ki67 in the liver tissue. The fluorescent TUNEL assay was performed to evaluate the hepatocyte apoptosis. The present study identified that the CBD-VEGF injection could significantly promote vascularization of the liver tissue of fibrotic mice and attenuate liver fibrosis. Furthermore, hepatocyte apoptosis and hepatic stellate cell activation were attenuated by CBD-VEGF treatment. CBD-VEGF treatment could additionally promote hepatocyte regeneration in the liver tissue of fibrotic mice. Thus, it was suggested that CBD-VEGF may be used as a novel therapeutic intervention for liver fibrosis.

Sepsis is one of the most fatal inflammatory diseases with multiple organ failure caused by pathological infection. α-Hederin, a monodesmosidic triterpenoid saponin, has many biological activities including anti-inflammation. This study aimed to investigate the effect of α-Hederin on lung and liver injuries in septic mice. Mice underwent cecal ligation and puncture-induced sepsis were intraperitoneally injected with 0.3 or 3 mg/kg α-Hederin. α-Hederin treatment dose-dependently attenuated the lung and liver injuries in septic mice. Correspondingly, α-Hederin significantly decreased malondialdehyde production, increased the levels of superoxide dismutase and glutathione in lung tissues, reduced serum alanine aminotransferase and aspartate aminotransferase activities, and suppressed the levels of TNF-α and IL-6 in both tissues and in the serum. Moreover, α-Hederin augmented CD206 level and inhibited the productions of CD86 and iNOS in lung and liver tissues of septic mice. Importantly, p-p65/p65 was suppressed, whereas IκB was elevated by α-Hederin. In conclusion, α-Hederin could improve the lung and liver injuries in mice with sepsis by regulating macrophage M1/M2 polarization and inhibiting the activation of NF-κB signaling pathway.

Mice lacking the intracellular glucocorticoid‐regenerating enzyme 11β‐hydroxysteroid dehydrogenase type 1 (11β‐HSD1) are protected from age‐related spatial memory deficits. 11β‐HSD1 is expressed predominantly in the brain, liver and adipose tissue. Reduced glucocorticoid levels in the brain in the absence of 11β‐HSD1 may underlie the improved memory in aged 11β‐HSD1 deficient mice. However, the improved glucose tolerance, insulin sensitisation and cardioprotective lipid profile associated with reduced peripheral glucocorticoid regeneration may potentially contribute to the cognitive phenotype of aged 11β‐HSD1 deficient mice. In the present study, transgenic mice with forebrain‐specific overexpression of 11β‐HSD1 (Tg) were intercrossed with global 11β‐HSD1 knockout mice (HSD1KO) to examine the influence of forebrain and peripheral 11β‐HSD1 activity on spatial memory in aged mice. Transgene‐mediated delivery of 11β‐HSD1 to the hippocampus and cortex of aged HSD1KO mice reversed the improved spatial memory retention in the Y‐maze but not spatial learning in the watermaze. Brain‐derived neurotrophic factor (BDNF) mRNA levels in the hippocampus of aged HSD1KO mice were increased compared to aged wild‐type mice. Rescue of forebrain 11β‐HSD1 reduced BDNF mRNA in aged HSD1KO mice to levels comparable to aged wild‐type mice. These findings indicate that 11β‐HSD1 regenerated glucocorticoids in the forebrain and decreased levels of BDNF mRNA in the hippocampus play a role in spatial memory deficits in aged wild‐type mice, although 11β‐HSD1 activity in peripheral tissues may also contribute to spatial learning impairments in aged mice.