• Journal of Veterinary Science
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• v.21 no.5
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• pp.78.1-78.15
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• 2020

Background: Enteritis is one of the most frequently reported symptoms in piglets infected with porcine circovirus type 2 (PCV2), but the immunopathogenesis has not been reported. Objectives: This study examined the effect of a PCV2 infection on the intestinal mucosal immune function through morphological observations and immune-related molecular detection. Methods: Morphological changes within the ileum of piglets during a PCV2 infection were observed. The expression of the related-molecules was analyzed using a gene chip. The immunocyte subsets were analyzed by flow cytometry. The secretory immunoglobulin A (SIgA) content was analyzed by enzyme-linked immunosorbent assay. Results: The PCV2 infection caused ileal villus damage, intestinal epithelial cells exfoliation, and an increase in lymphocytes in the lamina propria at 21 days post-infection. Differentially expressed genes occurred in the defense response, inflammatory response, and the complement and coagulation cascade reactions. Most of them were downregulated significantly at the induction site and upregulated at the effector site. The genes associated with SIgA production were downregulated significantly at the induction site. In contrast, the expression of the Toll-like receptor-related genes was upregulated significantly at the effector site. The frequencies of dendritic cells, B cells, and CD8⁺T cells were upregulated at the 2 sites. The SIgA content decreased significantly in the ileal mucosa. Conclusions: PCV2 infections can cause damage to the ileum that is associated with changes in immune-related gene expression, immune-related cell subsets, and SIgA production. These findings elucidated the molecular changes in the ileum after a PCV2 infection from the perspective of intestinal mucosal immunity, which provides insights into a further study for PCV2-induced enteritis.

• Journal of Biomedical Engineering Research
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• v.42 no.6
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• pp.251-258
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• 2021

Hyaluronic acid (HA) microspheres (MSs) crosslinked with polyethylene glycol diglycidyl ether (PEGDE) are prepared using a simple fluidic device (SFD) to investigate the optimized parameters. A solution mixture of PEGDE in 2-methyl-1-propanol was prepared as a continuous phase in SFD. HA solutions of 1 wt% concentration were introduced into SFD as a discontinuous phase. The HA solution prepared by stirring for more than 48 h exhibited spherical MSs at the needle tip inside the ring cap. As the flow rate of the continuous phase increased from 0.7 to 1.9 mL/min, the diameter of the MS decreased from 173±36 ㎛ to 129±13 ㎛. Although the PEGDE concentration in the range of 0.2 to 1.8 vol% did not affect the diameter of the MS, the microstructure of MS, consisting of inner hollow void and wall, was changed. The inner void and wall size decreased and increased from 79.5 ㎛ to 57.2 ㎛ and from 10.3 ㎛ to 21.4 ㎛, respectively, with increasing PEGDE concentration from 0.2 vol% to 1.8 vol%. FT-IR peaks located around 2867 cm^-1 and 1088 cm^-1 indicated that the HA MS prepared at different PEGDE concentrations were chemically crosslinked. The HA MSs containing different PEGDE concentrations exhibited quantitative cell viability of more than 98%. L-929 cells adhered well to the HA MSs and proliferated continuously with increasing culture time to 48 h regardless of PEGDE concentration, implying that the HA MSs are clinically safe and effective.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.39-39
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• 2001

Effects of Barodon$^{(R)}$ on body growth, histological changes in seminiferous tubules of testes and serum level of testosterone and FSH were examined in juvenile rats from 5 to 38 wks of age. All rats supplied feed and Barodon-added water(0, 0.1, 0.15, 0.3 and 1.0%) available ad libitum. DNA distribution of testicular cells from control rats obtained by flow cytometry was characterized by 4 peaks representing I : elongated, II round spermatids(1N), III: a variety of 2N cells and IV: 4N cells. Frequencies of 4 testicular cell types were calculated using cumulative DNA frequency distributions. Body growth from 18wk of age was significantly accelerated in rats supplied water with 0.15% Barodon compared to other groups. Testes weights tended to be greater in 0.15% Barodon-treated rats than in control, without significant difference. Diameter of seminiferous tubules advanced in 0.15-0.3% Barodon till 26 wk of age, but there was not significant different. Proportion of spermatids in seminiferous tubules was 48.4% at 6wk of age(round: 81.2% and elongated: 18.8% as proportion of total spermatids) and frequency of spermatids was higher in 0.3% Barodon group(57.1%) than in control(44.0%), but there was no significant difference. Serum testosterone of all groups significantly elevated at 18wk of age and level in 0.15% Barodon group was greatly higher than in others. Serum FSH at 10wks was greatly higher in 0.1-0.3% Barodon groups compared to control, but there were no significant differences. It is concluded that 0.15-0.3% Barodon treatment tended to induce precocious puberty in rats.

• Journal of Physiology & Pathology in Korean Medicine
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• v.36 no.6
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• pp.235-241
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• 2022

The purpose of this study was to investigate the immune activation effect of Sasamsaengmaek-san (SSSMS) consisted of a mixture of Adenophora triphylla var. japonica, Liriope platyphylla, Panax ginseng C. A. Meyer and Schisandra chinensis. in Balb/c mice. Measuring alanine aminotransferase (ALT) and aspartic acid transaminase (AST) levels in Balb/c mice was performed to analyze the cytotoxicity. Cytokines (IFN-γ, IL-2, IL-12) which regulate the immune activation in Balb/c mice were measured by enzyme-linked immunosorbent assay (ELISA). Activated T lymphocytes in peripheral blood mononuclear cell (PBMC), spleen, lymph nodes were analyzed by flow cytometry using percentages. All tests were compared with red ginseng 100 ㎍/mL (RG 100), which is the most used for immune activity. As a result, cytokine activity was significantly increased at SSSMS 300 group. Activated T lymphocytes in PBMC, spleen, lymph nodes were significantly increased at SSSMS 300 group. These results suggest that there is a possibility of SSSMS activating an immune system by activating the cytokines, and it is confirmed that SSSMS also effective for generation and differentiation of T, B lymphocytes which activate the immune response.

• The Korea Journal of Herbology
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• v.24 no.4
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• pp.77-86
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• 2009

Objectives : The proliferation and migration of human aortic smooth muscle cells (HASMC) in response to activation by various stimuli plays a critical role in the initiation and development of atherosclerosis. This study was conducted to examine the effects of Nelumbo nucifera leaves (NNL) on the proliferation and migration of HASMC. Additionally, the mechanisms involved in any observed effects were also evaluated. Methods : Apoptotic cells were measured by staining with FITC-labeled annexin V, followed by flow cytometric analysis. The expression level of apoptosis related proteins was confirmed by western blot. And MMP-9 activity was measured by gelatin zymography and MMP-9 expression was measured by ELISA Results : NNL completely inhibited the proliferation of HASMC via induction of the expression of apoptotic proteins including annexin V, cleaved poly ADP-ribose polymerase (PARP), and caspase-3 and -8. NNL treatment resulted in the release of cytochrome c into cytosol, a loss of mitochondrial membrane potential, a decrease in Bcl-2 and Bcl-xL and an increase in Bax expression. NNL also blocked HASMC migration via suppression of MMP-9. Conclusions : Taken together, these results indicate that NNL has the potential for use as an anti-artherosclerosis agent.

• The Korea Journal of Herbology
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• v.24 no.1
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• pp.79-88
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• 2009

Objectives : It has been recently shown that Piperis Longi Fructus (PLF) is involved in the reduction of eosinophil recruitment and production of Th2 cytokines in vivo. However, the main therapeutic mechanisms of PLF remains a matter of considerable debate. To investigate the therapeutic mechanisms of PLF, we examined the influence of PLF on regulatory T cells number, IgE, histamine production in vivo and Th1/Th2 cytokine balance in vitro. Methods : All mice were immunized on two different days (21 days and 7 days before inhalational exposure) by i.p. injections of 0.2 $m\ell$ alum-precipitated Ag containing 100 ${\mu}g$ of OVA bound to 4 mg of aluminum hydroxide in PBS. Seven days after the second sensitization, mice were exposed to aerosolized ovalbumin for 30 min/day on 3 days/week for 12 weeks(at a flow rate of 250 L/min, 2.5％ ovalbumin in normal saline) and PLF (150 mg/kg) were orally administered 3 times a week for 8 weeks. Splenocytes from C57BL/6 mice at 8 weeks of age were stimulated with anti-CD3 (1 mg/ml) plus anti-CD28 (1 mg/ml) antibody for 48hrs. IL-4 and IFN-$\gamma$ in the culture supernatants were measured by ELISA Results : The suppressive effects of PLF on asthma model were demonstrated by the increase the number of regulatory T cells and by reducing IgE, histamine production in vivo and modulation of Th1/Th2 cytokine balance. Conclusions : These results indicate that PLF has a deep inhibitory effects on asthma model mice by increase the number of regulatory T cells, and by reducing IgE, histamine production.

• Journal of Veterinary Science
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• v.24 no.3
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• pp.38.1-38.12
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• 2023

Background: Poor disease management and irregular vector control could predispose sheltered animals to disease such as feline Bartonella infection, a vector-borne zoonotic disease primarily caused by Bartonella henselae. Objectives: This study investigated the status of Bartonella infection in cats from eight (n = 8) shelters by molecular and serological approaches, profiling the CD4:CD8 ratio and the risk factors associated with Bartonella infection in shelter cats. Methods: Bartonella deoxyribonucleic acid (DNA) was detected through polymerase chain reaction (PCR) targeting 16S-23S rRNA internal transcribed spacer gene, followed by DNA sequencing. Bartonella IgM and IgG antibody titre, CD4 and CD8 profiles were detected using indirect immunofluorescence assay and flow cytometric analysis, respectively. Results: B. henselae was detected through PCR and sequencing in 1.0% (1/101) oral swab and 2.0% (1/50) cat fleas, while another 3/50 cat fleas carried B. clarridgeiae. Only 18/101 cats were seronegative against B. henselae, whereas 30.7% (31/101) cats were positive for both IgM and IgG, 8% (18/101) cats had IgM, and 33.7% (34/101) cats had IgG antibody only. None of the eight shelters sampled had Bartonella antibody-free cats. Although abnormal CD4:CD8 ratio was observed in 48/83 seropositive cats, flea infestation was the only significant risk factor observed in this study. Conclusions: The present study provides the first comparison on the Bartonella spp. antigen, antibody status and CD4:CD8 ratio among shelter cats. The high B. henselae seropositivity among shelter cats presumably due to significant flea infestation triggers an alarm of whether the infection could go undetectable and its potential transmission to humans.

• The Korean Journal of Physiology and Pharmacology
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• v.27 no.4
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• pp.383-398
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• 2023

Dihydroaustrasulfone alcohol (DA), the synthetic precursor of a natural compound (austrasulfone) isolated from the coral species Cladiella australis, has shown cytotoxic effects against cancer cells. However, it is unknown whether DA has antitumor effects on nasopharyngeal carcinoma (NPC). In this study, we determined the antitumor effects of DA and investigated its mechanism of action on human NPC cells. The MTT assay was used to determine the cytotoxic effect of DA. Subsequently, apoptosis and reactive oxygen species (ROS) analyses were performed by using flow cytometry. Apoptotic and PI3K/AKT pathway-related protein expression was determined using Western blotting. We found that DA significantly reduced the viability of NPC-39 cells and determined that apoptosis was involved in DA-induced cell death. The activity of caspase-9, caspase-8, caspase-3, and PARP induced by DA suggested caspase-mediated apoptosis in DA-treated NPC-39 cells. Apoptosis-associated proteins (DR4, DR5, FAS) in extrinsic pathways were also elevated by DA. The enhanced expression of proapoptotic Bax and decreased expression of antiapoptotic BCL-2 suggested that DA mediated mitochondrial apoptosis. DA reduced the expression of pPI3K and p-AKT in NPC-39 cells. DA also reduced apoptosis after introducing an active AKT cDNA, indicating that DA could block the PI3K/AKT pathway from being activated. DA increased intracellular ROS, but N-acetylcysteine (NAC), a ROS scavenger, reduced DA-induced cytotoxicity. NAC also reversed the chances in pPI3K/AKT expression and reduced DA-induced apoptosis. These findings suggest that ROS-mediates DA-induced apoptosis and PI3K/AKT signaling inactivation in human NPC cells.

• Journal of Microbiology and Biotechnology
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• v.33 no.10
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• pp.1384-1389
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• 2023

This work aimed to evaluate the feasibility of biohydrogen production from Barley Straw and Miscanthus. The primary obstacle in plant biomass decomposition is the recalcitrance of the biomass itself. Plant cell walls consist of cellulose, hemicellulose, and lignin, which make the plant robust to decomposition. However, the hyperthermophilic bacterium, Caldicellulosiruptor bescii, can efficiently utilize lignocellulosic feedstocks (Barley Straw and Miscanthus) for energy production, and C. bescii can now be metabolically engineered or isolated to produce more hydrogen and other biochemicals. In the present study, two strains, C. bescii JWCB001 (wild-type) and JWCB018 (ΔpyrFA Δldh ΔcbeI), were tested for their ability to increase hydrogen production from Barley Straw and Miscanthus. The JWCB018 resulted in a redirection of carbon and electron (carried by NADH) flow from lactate production to acetate and hydrogen production. JWCB018 produced ~54% and 63% more acetate and hydrogen from Barley Straw, respectively than its wild-type counterpart, JWCB001. Also, 25% more hydrogen from Miscanthus was obtained by the JWCB018 strain with 33% more acetate relative to JWCB001. It was supported that the engineered C. bescii, such as the JWCB018, can be a parental strain to get more hydrogen and other biochemicals from various biomass.

• Journal of dental hygiene science
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• v.23 no.4
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• pp.264-270
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• 2023

Background: Resin-based dental materials release residual monomers or other substances from incomplete polymerization into the oral cavity, thereby causing adverse biological effects on oral tissue. 10-Methacryloyloxydecyl dihydrogen phosphate (10-MDP), an acidic monomer containing dihydrogen phosphate and methacrylate groups, is the most commonly used component of resin-based dental materials, such as restorative composite resins, dentin adhesives, and resin cements. Although previous studies have reported the cytotoxicity and biocompatibility in various cultured cells, the effects of resin monomers on cellular aging have not been reported to date. Therefore, this study aimed to investigate the effects of the resin monomer 10-MDP on cellular senescence and inflamm-aging in vitro. Methods: After stimulation with 10-MDP, MC3T3-E1 osteoblast-like cells were examined for cell viability by WST-8 assay and reactive oxygen species (ROS) production by flow cytometry. The protein and mRNA levels of molecular markers of aging were determined by western blotting and RT-PCR analysis, respectively. Results: Treatment with 0.05 to 1 mM 10-MDP for 24 hours reduced the survival of MC3T3-E1 cells in a concentration-dependent manner. The intracellular ROS levels in the 10-MDP-treated experimental group were significantly higher than those in the control group. 10-MDP at a concentration of 0.1 mM increased p53, p16, and p21 protein levels. Additionally, an aging pattern was observed with blue staining due to intracellular senescence-associated beta-galactosidase activity. Treatment with 10-MDP increased the levels of tumor necrosis factor-α, interleukin (IL)-1β, IL-6 and IL-8, however their expression was decreased by mitogen-activated-protein-kinase (MAPK) inhibitors. Conclusion: Taken together, these results suggest that the exposure of osteoblast-like cells to the dental resin monomer 10-MDP, increases the level of cellular senescence and the inflammatory response is mediated by the MAPK pathway.