• Reproductive and Developmental Biology
• /
• v.33 no.2
• /
• pp.85-91
• /
• 2009

In this study, in vitro maturation system using fetal bovine serum (FBS) or porcine follicular fluid (pFF) was investigated to produce comparable oocytes to those derived from in vivo. Control group of oocytes was cultured in TCM 199 supplemented with 0.1% polyvinyl alcohol (PVA). Other three groups of oocytes were cultured in TCM 199 supplemented with 10% FBS, 10% pFF or 5% FBS + 5% pFF, respectively. After 44 h maturation, oocytes with the first polar body were activated with two electric pulses (DC) of 1.2 kv/cm for 30 ${\mu}sec$. Also, matured oocytes of four groups were reconstructed and fused. Reconstructed embryos were cultured in PZM-3 under 5% $CO_2$ in air at $38.5^{\circ}C$ for 6 days. The oocytes matured in the medium supplemented with FBS or/and pFF showed significantly higher maturation rates (64.0 vs. 73.9 to 85.2%). In PA embryos, cleavage rates (89.7 vs. 77.1 to 86.6%) and blastocysts rates (30.0 vs. 16.2 to 26.2%) were significantly higher in pFF group (p<0.05). In NT embryos, there was no difference among treatments in cleavage rate, but the blastocyst rates (28.5 vs. 15.5 to 24.6%) were significantly higher in pFF group (p<0.05). The apoptosis rate was significantly higher (p<0.05) in the control than other groups (10.8 vs. 4.9 to 8.2% for PA, 3.1 vs. 0.5 to 1.3% for NT). In order to select the comparable oocyte to in vivo oocytes, each group of oocytes was stained with Brilliant cresyl blue (BCB) after 42h maturation. The matured oocytes were separated according to color of cytoplasm; stained group (BCB+) and unstained group (BCB-). The oocytes matured in the presence of FBS or/and pFF showed significantly higher staining rates (70.3 to 72.7 vs. 35.1%) (p<0.05). To verify the fact that the supplementation of FBS or/and pFF can increase the maturation rates, cdc2 kinase activity, the catalytic subunit of MPF, was determined. The cdc2 kinase activity of the oocytes matured in the medium supplemented with FBS or/and pFF was significantly higher than control group (6.7 to 9.3 vs. 3.8). In conclusion, the supplementation of FBS or/and pFF can support in vitro maturation rate of porcine oocytes through the increment of cdc2 kinase activity level in the cytoplasm.

• The Journal of Pediatrics of Korean Medicine
• /
• v.30 no.3
• /
• pp.1-30
• /
• 2016

Objectives Previous studies have found out that Forsythiae Fructus (FF) extracts have anti-atopic activities by in vitro experiment. In order to understand more about FF extracts' benefit, we subdivided FF extracts depending on systematic fractionation method by using Methylene chloride (MC), Ethyl acetate (EtOAc), n-BuOH and n-hexane (n-Hx). This study is designed to examine the effect of FF fractions on the PMA- ionomycin-induced activation of RBL-2H3 mast cell lines in vitro and on the DNCB-induced activation of NC/Nga mice in vivo. Methods For this study, we examined IL-4, IL-13 production by ELISA analysis, IL-4, IL-13, IL-31, IL-31RA and TNF-${\alpha}$ mRNA expression by real-time PCR and manifestations of AP-1 and MAPKs transcription factors by western blotting in vitro. Through in vitro experiment, we selected FF n-BuOH fraction that seems the best effective in atopic dermatitis then induced it on NC/Nga mice by DNCB. We measured mice's WBC, eosinophil and neutrophil in heart blood, IL-4, IL-5, IFN-${\gamma}$ in the spleenocyte culture supernatant, the absolute cell numbers of CD4+, CD8+, B220+CD23+, CD3+CD69+ and Gr-1+CD11b+ in the PBMCs, ALN and dorsal skin, IL-5, IL-13, IL-31, IL-31RA in the dorsal skin by real-time PCR and the distribution of immune cells by H&E on dorsal skin and ANL and toluidine blue staining on dorsal skin. Results FF n-BuOH fraction suppressed IL-4, IL-13 production and mRNA expression of IL-4, IL-13, IL-31, IL-31RA and TNF-${\alpha}$. Results from the western blot analysis showed that FF n-BuOH fraction reduced the activation of the mast cell specific transduction factors involved in AP-1 by suppressing JNK and ERK phosphorylation. In the gross, atopic dermatitis induced by DNCB in NC/Nga mice were improved by oral administration of FF n-BuOH fraction. Oral FF n-BuOH fraction also decreased the level of IgE in mice's serum and the level of IL-4 and IL-5 in the spleenocyte culture supernatant, cell numbers of CD8+, B220+CD23+ in the PBMCs, CD4+ in the ALN and CD4+, Gr-1+CD11b+ in the dorsal skin and suppressed mRNA expression of IL-5, IL-13, IL-31, IL-31RA in the dorsal skin. Histological examination showed that infiltration levels of immune cells in atopic dermatitis induced NC/Nga mice were improved by FF n-BuOH fraction. Conclusions FF n-BuOH fraction can reduce pruritus by suppressing IL-31, IL-31RA secretion and modulate molecular mediators and immune cells associated with atopic dermatitis induced in NC/Nga mice which may have played a significant role in recovering atopic dermatitis symptoms.

• Applied Biological Chemistry
• /
• v.46 no.3
• /
• pp.165-170
• /
• 2003

The antioxidative activity of antioxidative substances produced from several bacterial strains isolated from fermented foods were tested by $DPPH\;({\alpha},{\alpha}'-diphenyl-{\beta}-picrylhydrazyl)$ free radical scavenging activity. One of the strains showing the highest antioxidative activity was identified as Bacillus sp. based on the morphological, biochemical, physiological characteristics, and 16S rRNA sequence, and named FF-7. The most optimal medium condition for the production of antioxidative substance from Bacillus sp. FF-7 was 2% galactose as carbon source and l% tryptone as nitrogen source. The antioxidative substance produced from FF-7 in these cultural medium was also tested by in vitro experimental models, the peruxidation of linoleic acid and the peroxidation of rat tissues microsomes by using thiobarbituric acid (TBA) for assay of free malondialdehyde production. The antioxidative activity against lipid peroxidation of rat tissues microsomes was shown in the following order; brain 97.50% > heart 79.95% > kidney 77.84% > spleen 77.47% > testis 69.96% > liver 62.45%. The antioxidative substance produced from FF-7 on linoleic acid peroxidation by IBA method was effectively inhibited during four days, and 0.05% BHT (butylated hydroxytoluene) used comparative control was also effectively inhibited. Results showed that the highest antioxidative activity by DPPH method of antioxidative substance produced from Bacillus sp. FF-7 was obtained by supplementing 2% galactose as carton source and l% tryptone as nitrogen source in cultured medium, this substance effectively inhibited the formation of TBARS in brain microsome in vit개 system and in linoleic acid peroxidation.

• Reproductive and Developmental Biology
• /
• v.36 no.4
• /
• pp.249-254
• /
• 2012

This study aimed at investigating whether a porcine follicular fluid (pFF) supplementation positively affects the characteristics of donor cells and the developmental competence of porcine cloned embryos. Ear fibroblast cells (donor cell) from an Massachusetts General Hospital miniature pig were cultured in different culture methods: (1) Dulbecco's modified Eagle's medium (DMEM)+10% FBS (Control); (2) DMEM+0.5% FBS (SS); and (3) DMEM+10% FBS+10% pFF (pFF) for 72 h. In each conditioned medium, the concentrations of 4 amino acids (Thr, Glu, Pro, and Val) in the pFF group were significantly different from those in the control group (p<0.05 or p<0.01). The proliferation of the cells cultured in the SS group was significantly lower than that of the other treatment groups (p<0.01). The population of apoptotic and necrotic cells in the SS group was significantly higher than that of either the control or the pFF group (p<0.01). The number of embryos that cleaved (p<0.05) and developed into blastocysts (p<0.01) in the SS group was significantly lower than that of either the control or the pFF group. Compared to other groups, the blastocysts produced from the donor cells in the pFF group had higher total cells and lower apoptotic cells (p<0.05). It can be concluded that pFF supplementation in the donor cell culture medium positively affects cell death, cell cycle and quality of the cloned blastocyst.

• Journal of radiological science and technology
• /
• v.40 no.2
• /
• pp.297-302
• /
• 2017

In this study, the generation of photoneutrons between the 10 MV FF mode and the FFF mode was evaluated and the amount of photoneutrons generated by the 10 MV and 15 MV energy changes in the FFF mode was evaluated. The generated neutrons were evaluated at 13 measurement points and the KTEPC was used to collect the generated neutrons. 10 MV FF mode was measured at 10 MV FF mode and FFF mode at all measurement points. In the superior direction, 0.455mSv and 0.152mSv were the largest, and more than 33% optical neutron was generated in FF. 10 MV in FFF mode, 15 MV in 15 MV, and 0.402 mSv in the direction of Superior, and 6.9% in the direction.

• Journal of Oriental Neuropsychiatry
• /
• v.18 no.1
• /
• pp.185-196
• /
• 2007

Objective : This experimental Study was designed to investigate the effects of FOENICULI FRUCTUS(FF) on the change of inhibition lactate dehydrogenase(LDH) activity in neuronal cells and cytokines production in serum of cerebral ischemic rats. Method : FOENICULI FRUCTUS(FF)freeze dry powder and FF on the LDH activity in neuronal cells. Changes of FF on the physiological parameters(PaO2, PaCO2, MABP and HR) in crerbral ischemic rats. Effects of FF on the IL-1beta production, $TNF-{\alpha}$ production, $TGF-{\beta}$ production, and IL-10 in serum of cerebral ischemic rats. MCAO :. cytokines production of serum by drawing from femoral arterial blood after MCAO 1 hr. Reperfusion : cytokines production of serum by drawing from femoral arterial blood after reperfusion 1 hr. Results and Conclusion : 1. FF did not inhibit lactate dehydrogenase(LDH) activity in neuronal cells. 2. In serum by drawing from femoral arterial blood after middle cerebral arterial occlusion(MCAO) 1 hr and reperfusion 1 hr, sample group was significantly decreased $IL-l{\beta}$ production compared with control group 3. In serum by drawing from femoral arterial blood after MCAO 1 hr and reperfusion 1 hr, sample group was significantly decreased $TNF-{\alpha}$ production compared with control group. 4. In serum by drawing from femoral arterial blood after MCAO 1 hr and reperfusion 1 hr, sample group was significantly increased $TGF-{\beta}$ production compared with control group. 5. In serum by drawing from femoral arterial blood after reperfusion 1 hr, sample group was significantly increased IL-10 production compared with control group. This results were suggested that FF had inhibitive effect on the brain damage by inhibited LDH activity, $IL-l{\beta}$ and $TNF-{\alpha}$production, but accelerated $TGF-{\beta}$ production and IL-10 production.

• Asian-Australasian Journal of Animal Sciences
• /
• v.14 no.10
• /
• pp.1360-1366
• /
• 2001

The aims of this study were first to evaluate the effects of different levels (20, 40 and 100%) and sources (follicular size: large, >7 mm; medium, >5-7 mm; small, 3-5 mm) of porcine follicular fluid (pFF) on the in vitro maturation (IVM) of porcine oocytes, and the effects of fertilization treatments and different culture conditions on development of fertilized oocytes were also investigated. No differences in the maturation (63.6-76.6%) and cleavage (24.8-34.3%) rates were observed among the 20,40 and 100% pFF groups (p>0.05). The cleavage rates of oocytes cultured and fertilized in 40% and 100% pFF maturation media were significantly higher than those fertilized in m199-NBCS (51.0-61.2% vs. 12.8-31.8%. p<0.05), regardless of sources of the pFF. When oocytes were fertilized in m199-NBCS followed by culture in rabbit oviducts for 4 days, the cleavage rate in 40% pFF group was better than that in 100% pFF group (46.9% vs. 32.5%, p<0.05). Two oocytes recovered from the oviducts in the 40% pFF group developed to blastocysts after IVC. However, none developed to blastocysts when fertilized in the IVM medium after being transferred to rabbit oviducts. In conclusion, addition of pFF accompanied with gonadotropins (FSH, LH) in IVM medium enhanced maturation and cleavage rates of porcine oocytes. Direct addition of sperm suspension to IVM medium may be an alternative to simplify the fertilization procedures and to reduce the mechanical lesion during manipulation. Furthermore, rabbit oviducts provide a better environment for the in vitro fertilized oocyte developing to the morula and blastocyst stages.

• Applied Biological Chemistry
• /
• v.40 no.3
• /
• pp.238-242
• /
• 1997

Seeds of five radish varieties were soaked in water for three hours and cellulose was coated. The seeds were classified as three groups in UV light; non-fluorescent(NF), partly fluorescent(PF), and fully fluorescent(FF) seeds. Germination rate was less in the order of NF>PF>FF seeds. The seed coat structure of NF seeds was dense and showed round shape, while those of PF and FF seeds were wrinkled or ruptured. The cotyledon and hypocotyl of NF seeds were normal, while those of PF and FF seeds were dwarf and showed some scars in the cotyledons.

• The Korea Journal of Herbology
• /
• v.23 no.3
• /
• pp.93-102
• /
• 2008

Objectives : Forsythiae Fructus (Forsythia koreana Nakai) has been used anti-inflammatory, diuretics, antidote, and antibacterials in traditional herbal medicine. The present study is focused on the inhibitory effect of Forsythiae Fructus ethanol extract (FF-E) on the production of inflammatory mediators such as NO, iNOS and proinflammatory cytokines ($TNF-{\alpha}$, $IL-1{\beta}$ and IL-6) in LPS-stimulated BV-2 cells, a mouse microglial cell line, and investigated the scavenging activity of FF-E. Methods : BV-2 cells were pre-incubated with FF-E for 30 min and then stimulated with LPS (1 ${\mu}g/m{\ell}$) at indicated times. Cell toxicity of GCF was determined by MTT assay. The levels of NO, PGE2 and cytokines were measured by Griess assay and ELISA. The mRNA and protein expressions of iNOS and cytokines were determined by RT-PCR and Western blotting. Free radical scavenging activity of GCF was determined by DPPH assay in tube test. Results : FF-E significantly inhibited the excessive production of NO, $PGE_2$, $TNF-{\alpha}$, and $IL-1{\beta}$ in LPS-stimulated BV-2 cells. In addition, FF-E attenuated the mRNA and protein expressions of iNOS, and proinflammatory cytokines. FF-E also significantly scavenged the DPPH free radicals in a dose-dependent manner. Conclusions : These results indicate that FF-E exhibits anti-inflammatory property by suppressing the transcription of inflammatory mediator genes, suggesting the anti-inflammatory property of FF-E may make it useful as a therapeutic agent for the treatment of human neurodegenerative diseases.

• Korean Journal of Acupuncture
• /
• v.36 no.2
• /
• pp.115-126
• /
• 2019

Objectives : Osteoporosis is a condition characterized by low bone mass and increased bone fragility. It has become a major problem of senior citizens. The purpose of this study is to experiment the effect of water extract of Forsythiae Fructus (wFF) on osteoclast differentiation; and the other purpose is to examine the effect of wFF on osteoporosis in ovariectomized rat. Methods : To investigate the effect of wFF on osteoclast differentiation and activity, RAW 264.7 cells were used. The number of TRAP positive cell, TRAP activity, pit area, mRNA expression of makers (RANK, TRAP, CA II, CTK, MMP-9, NFATc1, c-Fos), protein expression of makers (NFATc1, c-Fos) were investigated. For in vivo study, 40 female Sprague-Dawley (SD) rats were induced osteoporosis by ovariectomy (OVX) and then tested for anti-osteoporosis effect by administration of wFF. Results : wFF suppressed osteoclatogenesis, TRAP activity and pit area formation. Moreover, wFF decreased the expression of master differentiation factors (NFATc1, c-Fos) and also reduced the osteoclastogenesis-related markers (TRAP, CA II, CTK, MMP-9). These suggest that wFF inhibit osteoclasts differentiation and bone resorption. In the OVX rat model, wFF inhibited decreasing of BMD and trabecular area. Conclusions : Forsythiae Fructus should be effective for osteoporosis prevention and treatment.