• The Korea Journal of Herbology
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• v.29 no.5
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• pp.65-73
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• 2014

Objectives : The purpose of this study is to prove the effect of Gamikyejakjimo-tang (ji$\bar{a}$w$\grave{e}$igu$\grave{i}$sh$\acute{a}$ozh$\bar{i}$m-t$\bar{a}$ng, GK) on rheumatoid arthritis. Methods : We checked viability and measured production of IL-$1{\beta}$, IL-6, IL-17, TNF-${\alpha}$ in RAW 264.7 cell after treat by GK. Then we measured rheumatoid arthritis index score of DBA/1 mice with rheumatoid arthritis induced by CIA after GK oral administration, checked IL-$1{\beta}$, IL-6, IL-17, TNF-${\alpha}$ and hs-CRP tests in serum. Also we were observed mRNA expression of IL-$1{\beta}$, IL-6, IL-17 and TNF-${\alpha}$ in spleen by RT-PCR. Results : GK showed cell viability of 100% or higher in all concentration in RAW 264.7 cells. GK inhibited LPS-induced productions of rheumatoid arthritis mediators cytokine in RAW 264.7cells. GK treated group showed improvement from rheumatoid arthritis at decreased the index score. Also, GK treated group decreased level in serum of IL-1b, IL-6, IL-17, TNF-a and hs-CRP tests by 31%, 35%, 20%, 57% and 58% respectively. Finally, GK treated group showed decrease expression of IL-$1{\beta}$, IL-6, IL-17 and TNF-${\alpha}$ mRNA in spleen by 46%, 51%, 25% and 42% respectively. Conclusions : In this study, in-vitro and in-vivo results observed rheumatoid arthritis factors cytokine of IL-$1{\beta}$, IL-6, IL-17 and TNF-${\alpha}$ decrease in RAW 264.7 cells, serum, mRNA expression. Also, GK showed decrease of inflammation figure in hs-CRP tests depending on effect of rheumatoid arthritis. Thus, these results can used as a effective drug of GK for rheumatoid arthritis.

• The Journal of Korean Obstetrics and Gynecology
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• v.19 no.4
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• pp.1-16
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• 2006

Purpose : The purpose of this research was to investigate the effects of Kamibojoongikkitang (KBT) on the immune cells in C57BL/6 mice. Methods : KBT (500mg/kg) was administerd p.o. once a day for 7 days. Results : KBT decreased the cell viability of thymocytes in vivo and in vitro system and decreased the cell viability of splenocytes in vivo, but increased the viability of splenocytes in vitro system. In addition, KBT did not affect the population of helper T (Th) cells and cytotoxic T (Tc) cells in thymocytes and decreased the population of T- and B-lymphocytes and the population of Th and Tc cells in splenocytes. Furthermore, KBT did not affect the production of ${\gamma}%-interferon and interleukin-4 in splenocytes. KBT increased the production of nitric oxide in vivo but decreased the production of nitric oxide in vitro system. KBT enhanced the phagocytic activity of peritoneal macrophages in vivo, but decreased the phagocytic activity in vitro. Conclusion : KBT has an inhibitory action on the specific immune response via decrease of the cell viability of thymocytes and splenocytes and has a potent action on the non-specific immunity via increase of phagocytic activity of peritoneal macrophages.

• Journal of Korean Neurosurgical Society
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• v.38 no.5
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• pp.366-374
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• 2005

Objective : Nitric oxide[NO] is implicated in a wide range of biological processes in tumors and is produced in glioma. To investigate the role of NO and its interaction with the tumoricidal effects of anticancer drugs, we study the antitumor activities of NO donors, with or without anticancer drugs, in human glioma cell lines. Methods : U87MG and U373MG cells were treated with the NO donors sodium nitroprusside[SNP] and S-nitroso-N-acetylpenicillamine[SNAP], alone or in combination with the anticancer drugs 1,3-bis[2-chloroethyl]-1-nitrosourea[BCNU] and cisplatin. Cell viability, cell proliferation, DNA fragmentation, nitrite level, and the expression of Bcl-2 and Bax were determined. Results : NO was markedly increased after treatment with SNP or SNAP; however, the addition of the anticancer drugs did not significantly affect NO production NO donors or anticancer drugs reduced glioma cell viability and, in combination, acted synergistically to further decrease cell viability in a dose- and time-dependent manner. Cell proliferation was inhibited and apoptosis were enhanced by combined treatment. Bax expression was increased by combined treatment, whereas Bcl-2 expression was reduced. The antitumor cytotoxicity of NO donors and anticancer drugs differed according to cell type. Conclusion : BCNU or cisplatin can inhibit cell viability and proliferation of glioma cells and can induce apoptosis. These effects are further enhanced by the addition of a NO donor which modulates the antitumor cytotoxicity of chemotherapy depending on cell type. Further biological, chemical, and toxicological studies of NO are required to clarify its mechanism of action in glioma.

• Journal of Embryo Transfer
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• v.31 no.4
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• pp.355-365
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• 2016

The cryopreservation of sperm has become the subject of research for successful artificial insemination technologies. Antifreeze proteins (AFPs), one of the factors necessary for effective cryopreservation, are derived from certain Antarctic organisms. These proteins decrease the freezing point of water within these organisms to below the temperature of the surrounding seawater to protect the organism from cold shock. Accordingly, a recent study found that AFPs can increase the motility and viability of spermatozoa during cryopreservation. To evaluate this relationship, we performed cryopreservation of boar sperm with AFPs produced in the Arctic yeast Leucosporidium sp. AFP expression system at four concentrations (0, 0.01, 0.1, and $1{\mu}g/ml$) and evaluated motility using computer assisted sperm analysis. DNA damage to boar spermatozoa was measured by the comet assay, and sperm membrane integrity and acrosome integrity were evaluated by flow cytometry. The results showed that motility was positively affected by the addition of AFP at each concentration except $1{\mu}g/ml$ (p<0.001). Although cryopreservation with AFP decreased the viability of the boar sperm using, the tail DNA analyses showed that there was no significant difference between the control and the addition of 0.1 or $0.01{\mu}g/ml$ AFP. In addition, the percentage of live sperm with intact acrosomes showed the least significant difference between the control and $0.1{\mu}g/ml$ AFP (p<0.05), but increased with $1{\mu}g/ml$ AFP (p<0.001). Our results indicate that the addition of AFP during boar sperm cryopreservation can improve viability and acrosome integrity after thawing.

• Reproductive and Developmental Biology
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• v.40 no.4
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• pp.33-37
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• 2016

This study was conducted to evaluate effect of ${\alpha}$-linolenic acid (ALA) and bovine serum albumin (BSA) on viability, acrosome reaction and mitochondrial intact in frozen-thawed boar sperm. The boar semen was collected by gloved-hand method and cryopreserved using freezing extender containing 3 ng/mL ALA and/or $20\;{\mu}g/mL$ BSA. Cryo-preserved boar sperms were thawed in $37^{\circ}C$ water-bath for 45 sec to analysis. Viability, acrosome reaction, and mitochondrial intact were analyzed using flow cytometry. In results, viability of frozen-thawed boar sperm was significantly higher in only ALA+BSA supplement group than control group (p<0.05), whereas there was no difference either in ALA or BSA supplement. However, acrosome reacted sperm in both of live and all sperm population were significantly decreased in all treatment groups than control (p<0.05). Interestingly, mitochondrial intact of boar sperm was enhanced in ALA and ALA+BSA groups compared with control (p<0.05). In this study, we showed that supplementation of ALA and BSA in freezing extender enhanced the sperm viability, mitochondrial intact and decrease acrosomal membrane damage. In conclusion, our findings suggest that quality of frozen-thawed sperm in mammalians could improve by using of ALA and BSA.

• Preventive Nutrition and Food Science
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• v.12 no.2
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• pp.74-78
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• 2007

Fucoidan, that is high-molecular-weight sulfated polysaccharides extracted from brown seaweeds has been shown to elicit various biological activities. Here, we investigated the effects of fucoidan on cell proliferation and nitric oxide (NO) production in neuronal blastoma cell (SH-SY5Y). In the present study, we demonstrated that fucoidan treatment resulted in increase of cell proliferation and NO production. When cells were treated with amyloid-${\beta}$ (A${\beta}$) in the absence or presence of fucoidan, fucoidan recovered the cell viability decreased by A${\beta}$ peptides. To further determine whether nitric oxide synthase (NOS) is involved in proliferative effect of fucoidan, cells were treated with NOS inhibitors in the absence or presence of fucoidan. Selective constitutive nitric oxide synthase (cNOS) inhibitor, diphenylene iodonium chloride (DPI), caused a decrease of cell viability, whereas cell viability was increased by specific inducible nitric oxide synthase (iNOS) inhibitor, S-methylisothiourea (SMT), in the fucoidan-untreated cells. Treatment with fucoidan inhibited the cell viability decreased in DPI-exposed cells. In contrast, fucoidan had no effect on cell growth in SMT-treated cells, indicating that cNOS may not play a role in the proliferation of fucoidan-treated cells. The present data suggest that fucoidan has proliferative and neuroprotective effects and these effects may be associated with iNOS.

• Journal of Periodontal and Implant Science
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• v.43 no.4
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• pp.168-176
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• 2013

Purpose: The purpose of the current study was to examine the effect of dexamethasone (Dex) at various concentrations on the apoptosis and mineralization of human periodontal ligament (hPDL) cells. Methods: hPDL cells were obtained from the mid-third of premolars extracted for orthodontic reasons, and a primary culture of hPDL cells was prepared using an explant technique. Groups of cells were divided according to the concentration of Dex (0, 1, 10, 100, and 1,000 nM). A 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was performed for evaluation of cellular viability, and alkaline phosphatase activity was examined for osteogenic differentiation of hPDL cells. Alizarin Red S staining was performed for observation of mineralization, and real-time polymerase chain reaction was performed for the evaluation of related genes. Results: Increasing the Dex concentration was found to reduce cellular viability, with an increase in alkaline phosphatase activity and mineralization. Within the range of Dex concentrations tested in this study, 100 nM of Dex was found to promote the most vigorous differentiation and mineralization of hPDL cells. Dex-induced osteogenic differentiation and mineralization was accompanied by an increase in the level of osteogenic and apoptosis-related genes and a reduction in the level of antiapoptotic genes. The decrease in hPDL cellular viability by glucocorticoid may be explained in part by the increased prevalence of cell apoptosis, as demonstrated by BAX expression and decreased expression of the antiapoptotic gene, Bcl-2. Conclusions: An increase in hPDL cell differentiation rather than cellular viability at an early stage is likely to be a key factor in glucocorticoid induced mineralization. In addition, apoptosis might play an important role in Dex-induced tissue regeneration; however, further study is needed for investigation of the precise mechanism.

• Animal Bioscience
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• v.35 no.5
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• pp.670-676
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• 2022

Objective: This study was conducted to examine influence of skimmed milk-based extender (SM), INRA 96 extender and BotuSemen Gold extender on parameters of stallions' ejaculate during storage. Methods: In this study, 14 stallions between 4 and 20 years of age were monitored. Total and progressive motility, viability and morphology of sperm were evaluated at time intervals of 24, 48, and 72 hours after collection. Results: The total motility, progressive motility, and values of sperm with normal morphology were significantly higher in the INRA 96 and BotuSemen Gold extenders than in the SM (p<0.01). The sperm viability differed significantly in all extenders (p<0.01). The highest value of sperm viability was in INRA 96 (64.69%±0.67%) and lowest in SM (59.70%±0.81%). The highest differences occurred at 72 hours of storage. Values of total motility, progressive motility and sperm viability decreased over time (p<0.01). In case of sperm morphology there was no statistically significant decrease between 48- and 72-hour time intervals. Conclusion: It can be concluded that the extenders with a chemically defined composition have shown better indicators of insemination capabilities in ejaculates than the SM. The BotuSemen Gold extender is a suitable alternative to the INRA 96, when used within 48 hours; after 72 hours of storage, however, the INRA 96 showed a higher share of viable spermatozoa.

• Animal Bioscience
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• v.37 no.2
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• pp.203-209
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• 2024

Objective: This study aimed to assess the impact of the dilution ratio of Tris diluent, storage at 0℃, and long-distance transportation on the spermatozoa of Simmental cattle. It also validated the feasibility of the regional distribution of fresh semen. Methods: In experiment 1, semen was diluted at four dilution ratios (1:6, 1:9, 1:12, and 1:15) to determine the optimal dilution ratio of Tris diluent. In experiment 2, we assessed sperm viability, progressive motility (objectively assessed by computer-assisted sperm analyzer), and acrosome intactness in Tris dilutions kept at constant 0℃ for 1, 3, 6, 9, and 12 days. We compared them to Tianshan livestock dilutions (Commercial diluent). In experiment 3, semen was diluted using Tris diluent, and sperm quality was measured before and after long-distance transport. Artificial insemination of 177 Simmental heifers compared to 156 using Tianshan Livestock dilution. Results: The outcomes demonstrated that 1:9 was the ideal Tris diluent dilution ratio. The sperm viability, Progressive Motility, and acrosome integrity of both Tris and Tianshan dilutions preserved at 0℃ gradually decreased over time. sperm viability was above 50% for both dilutions on d 9, with a flat rate of decline. The decrease in acrosome integrity rate was faster for Tianshan livestock dilutions than for Tris dilutions when stored at 0℃ for 1 to 6 days. There was no significant difference (p>0.05) in sperm viability between semen preserved in Tris diluent after long-distance transportation and semen preserved in resting condition. The conception rates for Tris dilution and Tianshan livestock dilution were 49.15% and 46.15% respectively, with no significant difference (p>0.05). Conclusion: This shows that Tris diluent is a good long-term protectant. It has been observed that fresh semen can be successfully preserved for long-distance transport when stored under 0℃ conditions. Additionally, it is feasible to distribute semen regionally.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.28 no.4
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• pp.277-286
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• 2006

In the present study, I investigated the effects of N-methyl-D-aspartate (NMDA), arachidonic acid (AA), and Nitric Oxide Synthase Inhibitor (NOS-I), alone or in combination, on the viability of cultured primary normal human oral keratinocytes (NHOK). Specifically, we examined whether AA and NOS-I could protect primary NHOK from glutamate cytotoxicity. The purpose of this study was therefore the preliminary study for the examination of the interaction between these agents and NHOK in order to elucidate the mechanisms by which epithelial growth and regeneration are regulated. NHOK were obtained from gingival tissue of 20 individuals aged 20 to 29, and third passage (P3) cells were used for this study. Cell viability was measured by the MTT assay. NMDA and NNA, a calcium dependent NOS inhibitor, induced an initial increase in cell number, which subsequently decreased by the $7^{th}$ day. Low concentration of AA ($0.5\;{\mu}M$ & $1\;{\mu}M$) induced an increase in cell number while high concentrations of AA ($5\;{\mu}M$ & $10\;{\mu}M$) induced a decrease in cell number. The decrease in cell number induced by NMDA at the $7^{th}$ day was abolished by the addition of low concentrations of AA ($0.5\;{\mu}M$ & $1\;{\mu}M$) or NOS inhibitors. Low concentrations of AA ($0.5\;{\mu}M$ & $1\;{\mu}M$) or NOS inhibitors may protect the NHOK from NMDA induced cytotoxicity. These reactions might be related to the NMDA receptor in the cell and the change of the intracellular calcium ion concentration.