• Journal of Veterinary Clinics
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• v.35 no.2
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• pp.39-45
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• 2018

We evaluated the effects of green tea extract (GTE) supplementation at different dilution steps on boar sperm freezing and in vitro fertilization. Sperm intracellular hydrogen peroxide ($H_2O_2$), motility, viability, acrosome integrity and morphology were determined. In addition, sperm IVF parameters (penetration and monospermy) and glutathione (GSH) levels of presumptive zygotes (PZs) were evaluated. Semen was diluted in lactose egg yolk (LEY) and cooled at $5^{\circ}C$ for 3 h (first dilution step) and then diluted in LEY with 9% glycerol and maintained at $5^{\circ}C$ for 30 min (second dilution step). Four experimental groups were compared: first and second dilution steps without GTE (control), first dilution step with GTE (Step 1), second dilution step with GTE (Step 2) and first and second dilution step with GTE (Step 1+2). The spermatozoa were frozen in nitrogen vapor. Higher sperm motility, viability and acrosome integrity after thawing were observed in Step 1, Step 2 and Step 1+2 groups compared with the control (P < 0.05). Lower $H_2O_2$ level was observed in Step 1+2 compared with control and Step 1 (P < 0.05). For IVF, matured oocytes were co-cultured with spermatozoa frozen according to the experimental groups. GSH levels of PZs were significantly higher in Step 2 and Step 1+2 than in control and Step 1 (P < 0.05) without a significant difference in IVF parameters. In conclusion, supplementation with GTE in both first and second dilution steps during the freezing process resulted in better boar sperm cryopreservation and might be beneficial for further embryo development.

• The Journal of Korean Obstetrics and Gynecology
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• v.19 no.3
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• pp.121-134
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• 2006

Purpose : Cuscuta chinensis Lam. is utilized extensively as important medicines for threatened abortion habitual abortion. However, objective data related to an embryo is not existed until now. Therefore, this study is focused to find out stability of Cuscuta chinensis Lam. about an embryo during pregnancy based on data related to stability of nerve cell and antioxydative effect by using neural cell line, PC 12 cell. Methods : Experimentation concerns cytotoxic effects and antioxydative effect through methods such as MTT ssay, western botting after abstracting an undiluted solution from domestic Cuscuta chinensis Lam. Results : 1. As a result of experimentation on MTT assay according to each magnification from Cuscuta chinensis Lam. extraction solution with different abstraction methods, cytotoxic effect is not observed to all extract except an undiluted solution which is abstracted from MeOH stiring. Also, an undiluted solution in stiring with MeOH could not confirm whether come from Cuscuta chinensis Lam. or not. 2. As a result of revelation of Bax and GSK-3${\beta}$ which is responsive to the first stage from general stress in order to observe antioxydative effects of Cuscuta chinensis Lam. revelation of Bax by Cuscuta chinensis Lam. appeared to decrease. Conclusion : Cytotoxic effects with Cuscuta chinensis Lam. about PC12 cell is not discovered and it assume that it would be anti apoptotic effect by ROS as Bax and GSK-3${\beta}$ inviable effect. In the future, this study could be used as basic data for additional research on Cuscuta chinensis Lam. and effect and stability of complicated prescriptions for keeping pregnancy.

• Herbal Formula Science
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• v.16 no.1
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• pp.65-78
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• 2008

Objectives : this study was to access the effect of Hyeonggaeyeongyotang water extracts, a polyherbal formula has been used as folk medicine, on the fertility and early embryonic development of male and female Wistar rats when administered by oral gavage. Methods : In male rats, Hyeonggaeyeongyotang extract were dosed 4 weeks before pairing and 2 weeks after mating including the mating periods up to termination after necropsy of the majority of the females. In female rats, they were dosed 2 weeks before pairing, and from Day 0 to Day 7 of gestation. This study was conducted in accordance with the recommendations of the KFDA Guideline [2005-60] for Detection of Toxicity to Reproduction for Medicinal Products. Results: 1. No Hyeonggaeyeongyotang extract treatment-related changes on the clinical signs and mortalities, the Food consumptions, the Body weights and gains were demonstrated in all dosed levels tested in this study except for 500ml/kg-dosing male group in which a significant(p<0.05) increase of body gains was detected during day 0-7 after dosing. 2. No Hyeonggaeyeongyotang extract treatment-related changes on the pre-coital intervals, the estrus cycles, the mating index, conception rate and fertility index were demonstrated in all dosed levels tested in this study. 3. No Hyeonggaeyeongyotang extract treatment-related gross findings on reproductive organs, the weights of reproductive organs, histopathological findings on reproductive organs, the corpora lutea number, implantation site number, live fetus number, number of resorpted embryo and pre-and post-implatation loss were demonstrated in all dosed levels tested in this study. Conclusions : Base on the results, it is considered that the NOAEL (No-Observed-Adverse-Effect Level) for fertility and early embryonic development toxicity of Hyeonggaeyeongyotang extract was under 2000ml/kg/day in Wistar male and female rats because there no treatment-related changes on the fertility and early embryonic developmental index were demonstrated in all dosed levels tested.

• Archives of Pharmacal Research
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• v.18 no.5
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• pp.332-335
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• 1995

The angiogenic activity of Aloe vera (Aloe baradensis), known as a good healing plant, was investigated. We have extracted and fractionated dichloromethane extract (G1M1D1) and methanol soluble fraction of dichloromethane extract (G1M1D1M1) which contain low-molecular weight substances of Aloe vera gel. G1M1D1 and G1D1M1 fractions induced a radially arranged, spoke-sheel-like vasculature in chick embryo chorioallantoic membrane (CAM) assay. The angiogenic activity was dose-dependent and the angiogenic pattern in the CAM assay. The angiogenic activity was dose-dependent and the angiogenic pattern in the CAM assay was very similar to that of phorbol 12-myristate-13-acetate (PMA) used as a positive control. The modified CAM assay, e simple and accurate quantitating method, was used to quntitate the angiogenic activity of G1D1M1 fraction. Application of G1M1D1M1 fraction ($100\mug/egg$) resulted in much more intense angiogenesis than in control while slightly less intense angiogenesis than in PMA (100 ng/egg).

• The Journal of Korean Obstetrics and Gynecology
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• v.34 no.4
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• pp.30-45
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• 2021

Objectives: This study was performed to investigate the effects of pregnancy-related four herbal medicines (Samul-tang, Onkyung-tang, Chokyungjongok-tang and Moutan Radicis Cortex) on the endometrial and placental cells. Methods: In this study, we examined viability and decidualization of telomerase immortalized human endometrial stromal cell lines (T-HESCs) and viability and invasion ability of human first trimester trophoblast cell lines Sw.71 by four herbal medicines (Samul-tang, Onkyung-tang, Chokyungjongok-tang and Moutan Radicis Cortex) Results: In the study, we showed that Samul-tang, Onkyung-tang, Chokyungjongok-tang increased decidualization marker prolactin (PRL) and insulin-like growth factor-binding protein 1 (IGFBP1) in T-HESCs. Moutan Radicis Cortex decreased the mRNA level of PRL and IGFBP1, and the protein level of PRL and IGFBP1 had no significant effect. Moreover, four herbal medicines reduced invasion ability of Sw.71 cells. Conclusions: These results suggest that Samul-tang, Onkyung-tang, and Chokyungjongok-tang have beneficial effects on successful embryo implantation and pregnancy maintenance by increasing decidualization markers such as PRL and IGFBP1. Moutan Radicis Cortex reduces the mRNA levels of PRL and IGFBP1, which may adversely affect pregnancy. Further investigations are needed to elucidate the significance of the decreased invasive ability of Sw.71 cells induced by four herbal medications.

• Toxicological Research
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• v.18 no.2
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• pp.117-127
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• 2002

Saururus chinensis Baill has been used in Korean folk medicine for the treatment of various diseases such as edema, Jaundice, and furuncle. The components of this plant were extracted into four fraction. Among the four fraction, hexane and ethyl acetate fraction were highly toxic to 3T3 mouse embryo fibroblast and Raw 264.7 mouse macrophage, but n-butanol and residue fraction did not show any toxic effect to those cell lines. n-Butanol and residue fraction exhibited antioxidant effects on hydro-gen peroxide, hydroxyl radical, and superoxide anion directly in vitro and in the 3T3 fibroblasts. All the four fractions inhibited lipid peroxidation measured by thiobarbituric acid reactive substances (TBARS) formation. In addition, n-butanol and residue fraction showed inhibitory effects on lipopolysaccharide (LPS)-induced nitric oxide production, and also down-regulated inducible nitric oxide synthase (iNOS) mRNA transcription 6 h after LPS stimulation in Raw 204.7 cells. Only n-butanol fraction, which mainly consists of flavonoids, inhibited NF-kB activation by decreasing IkBa degradation 90 min after LPS stimulation. horn the results, it is suggested that this plant could be a good candidate material for drug development based on its antioxidant and/or anti-inflammatory constituents.

• Fisheries and Aquatic Sciences
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• v.16 no.3
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• pp.177-185
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• 2013

The development of species-specific fish cell lines has become a valuable tool for biological research. In recent years, marine medaka Oryzias dancena has been recognized as a good experimental model fish but there are no reports of establishment of cell lines from this fish. In this study, two cell lines from O. dancena blastula embryos were established from 41 total trials (4.9%). The two cell lines displayed typical in vitro morphology and have been cultured for >121 passages, which corresponds to 293 days. The doubling times of the cell lines were 29.84 and 28.59 h, respectively, and both possessed the potential to expand in a clonal manner, albeit with significant differences between the two cell lines. The absence of any of the four main medium supplements; i.e., fish serum, fetal bovine serum, basic fibroblast growth factor, and medaka embryo extract, significantly inhibited growth. The proportion of cells possessing normal chromosome number was 45% and 46.7% of the cell lines, respectively. Taken together, two cell lines that proliferate continuously were established from marine medaka and these cell lines may provide a basic tool for characterizing the unique features of this fish species.

• KSBB Journal
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• v.21 no.4
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• pp.255-259
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• 2006

The biodegradable hyaluronic acid(HA) membranes cross-linked with lactide using the crosslinking agent, 1-ethyl-3(3-dimethyl aminopropyl) carbodiimide(EDC) were prepared as a potential biocompatible material for tissue engineering. HA membranes having different mechanical properties were synthesised by varying degree of the mole ratio of lactide to HA, EDC concentration, and crosslinking temperature. HA membranes were degradable in water solution and the degradation became slower with the increasing mole ratio of lactide to HA. HA membranes were sterilized using ethylene oxide gas and extracted with cell culture medium for 24 h at $37^{\circ}C$ and 200 rpm. Cytotoxicity of the extract was tested using NIH/3T3 mouse embryo fibroblast as a model cell. Growth inhibition was not observed in the extracts of HA membranes with the mole ratios of lactide to HA, 5 or 10, and 10% EDC concentration, however 11% of growth inhibition was observed in the extract with the mole ratio of 13. Growth inhibition was not observed in the extracts of HA membranes prepared with 5% EDC or 10% EDC and the mole ratio of lactide to HA, 10, however 12% of growth inhibition was observed in the extract with 20% EDC. Cytotoxicity was not observed in the extracts of HA membranes prepared at varying crosslinking temperatures, $15^{\circ}C,\;25^{\circ}C,\;and\;28^{\circ}C$ with the mole ratio of lactide to HA, 10 and 10% EDC.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.45 no.3
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• pp.255-263
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• 2019

Isomaltol glycoside is a hydrophilic furanic glycoside in which the amino acids and sugars of ginseng are thermally denatured during red ginseng production. Various skin whitening tests were conducted on isomaltol glycoside containing a lot of red ginseng extract in order to investigate the skin whitening effect as a cosmetic raw material. We have tested melanin content assay in B16-F10 cells, zebrafish embryo pigmentation assay, mushroom tyrosinase inhibitory activity, western blot analysis to determine skin whitening activity of isomaltol glycosides. In the zebrafish melanin content assay, isomaltol glycoside decreased total melanin content by about 20% and zebrafish tyrosinase activity by about 10% after treatment with 50 and $100{\mu}g/mL$ compared to the untreated control group. Isomaltol glycoside also showed a concentration-dependent decrease in melanin content in B16-F10 melanoma. Furthermore, it increased the expression of MITF phosphorylation factors p-AKT and p-ERK in B16-F10 melanoma and decreased the concentration of MITF. It also inhibited tyrosinase, TRP-1 and TRP-2 expression. The content of isomaltol glycoside was about 3% in the ginseng extract and about 1% in the ginseng root. Thus, isomaltol glycoside is considered as one of the main components that exhibit the whitening activity of ginseng when considered quantitatively as whitening activity.

• Journal of Life Science
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• v.28 no.10
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• pp.1156-1162
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• 2018

To investigate the anti-thrombotic activities of the lotus (Nelumbo nucifera Gaertner), various hot-water extracts were prepared from the leaf, pod of seed (PS), seed, embryo of seed (ES), root, and the node of root (NR) of the lotus. The highest extraction ratio was found in the NR (20.3%), followed by the seed, root, leaf, ES, and PS. These extracts had pH and acidity levels ranging from 5.6~6.5 and 0.06~0.20%, respectively. The seed extract showed 70% brix, whereas the leaf and PS extracts showed less than 0.1% brix. The highest contents of total polyphenol (179.7 mg/g), total flavonoids (161.4 mg/g), and reducing sugar (161.4 mg/g) were observed in the leaf extract, and the highest total sugar content (873.0 mg/g) in the seed extract. Anti-coagulation assay of the extracts of NR, leaf, and PS showed strong activities. In particular, at a concentration of 5 mg/ml, the PS extract had 15-fold extended thrombin, prothrombin, and activated partial thromboplastin times. However, only the ES extract showed activities inhibitory to platelet aggregation, with treatment with 0.25 mg/ml of ES extract decreasing platelet aggregation to 25.1%, a reduction comparable to that effected by aspirin. The extracts other than the seed extract showed no hemolysis activities against human RBC at treatments of up to 1 mg/ml. These results suggest that the NR, PS, seed, and ES, all byproducts of the lotus agriculture industry, have high potential as novel sources of anti-thrombotic agent.