• Clinical and Experimental Reproductive Medicine
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• v.24 no.2
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• pp.153-165
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• 1997

In male reproducible health, fertility and IVF (in-vitro fertilization), semen analysis has been most important. Semen analysis can be divided into concentration, motional and morphological analysis of sperm. The existing method which was developed earlier to analyze semen concentrated on the sperm motility analysis. To provide more useful and precise solutions for clinical problems such as infertility, semen analysis must include sperm morphological analysis. But the traditional tools for semen analysis are subjective, imprecise, inaccurate, difficult to standardize, and difficult to reproduce. Therefore, with the help of development of microcomputers and image processing techniques, we developed a new sperm morphology analyzer to overcome these problems. In this study the agreement on percent normal morphology was studied between different observers and a computerized sperm morphology analyzer on a slide-by-slide basis using strict criteria. Slides from 30 different patients from the SNUH andrology laboratory were selected randomly. Microscopic fields and sperm cells were chosen randomly and percent normal morphology was recorded. The ability of sperm morphology analyzer to repeat the same reading for normal and abnormal cells was studied. The results showed that there was no significant bias between two experienced observers. The limits of agreement were 4.1%${\sim}$-3.8%. The Pearson correlation coefficient between readers was 0.79. Between the manual and sperm morphology analyzer, the same findings were reported. In this experiments the slides were stained by two different methods, PAP and Diff-Quik staining methods. The limits of agreement were 7.2%${\sim}$-5.7% and 6.0%${\sim}$-6.3%, respectively. The Pearson correlation coefficients ware 0.76 and 0.91, respectively. The limits of agreement was tighter below 20% normal forms. In the experiments of repeatability, 52 cells stained by PAP and Diff-Quik staining methods were analyzed three times in succession. Estimating pairwise agreement, the kappa statistic for the pairs were 0.76, 0.81, 0.86, and 0.75, 0.88, 0.88 respectively. In this study it was shown that there was good agreement between manual and computerized assessment of normal and abnormal cells. The repeatability and agreement per slide of computerized sperm morphology analyzer was excellent. The computer's ability to classify normal morphology per slide is promising. Based on results obtained, this system can be of clinical value both in andrology laboratories and IVF units.

• Korean Journal of Plant Resources
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• v.21 no.3
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• pp.184-191
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• 2008

This experiments were carried out to find out the effects of different explant materials, kinds and concentration of plant growth regulators, and total nitrogen and sucrose contents on the in vitro regeneration of Abeliophyllum distichum Nakai. The effects of growth regulators on regeneration from 3 explant sources (leaf, internode and node) were more or less same. Leaf explants produced only callus with 2ip (Isopentenyladenine) and NAA (Naphthaleneacetic acid) treatment and other regulators had no effects. Test with internode explants yielded about same results but callus was obtained with 2,4-D (2,4-Dichlorophenoxyacetic acid). Node explants resulted in shoot regeneration by all regulator treatment except NAA and 2,4-D, but control also showed similar results. Callus formation from internode and node explants was vigorous by 2ip, zeatin, and 2,4-D treatments and high NAA concentration resulted in higher callus formation. In this experiment, various mixed treatment of growth regulators were also employed, using node as explant material. Shoot regeneration was obtained with BA (Benzyl adenine) + NAA treatments but the results were comparable with control. Generally shoot and root regeneration was poor with all combined treatment except 2ip + NAA and 2,4-D + NAA. However, callus was formed readily with all treatments. In this experiment, combined treatments of regulators were applied on the callus derived from singular regulator treatment. The results showed no shoot and root regeneration with any combination of 2,4-D, IAA (Indoleacetic acid) and NAA, but soft milky white callus was formed in all the treatments. No shoot and root regeneration was observed with any combination of 2iP, NAA and IAA, but somewhat hard, light green callus was formed in all the treatments. Callus formation decreased with high kinetin concentration in case of kinetin + NAA treatment. The experiments with total nitrogen content of media showed that low concentrations of 15 and 30mM were effective for the shoot and root regeneration. Sucrose experiment demonstrated shoot regeneration with 1${\sim}$4% concentration, and root and callus formation with 2${\sim}$4%. No root and callus formation was observed with 0 and 1% sucrose.

• The Korea Journal of Herbology
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• v.23 no.2
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• pp.111-121
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• 2008

Objectives : The present study was carried out to investigate the effects of Astragali radix on the asthma treatment. EAR-I is an extract of astragali radix cultured for one year and EAR-III is an extract of astragali radix cultured for three years. Methods : Two asthma-induced groups mice group was treated respectively with EAR- I and EAR- III. The other group was not. Each group was analyzed in vivo and in vitro experiments. Results : In asthma-induced mice by OVA treatment, the weight of lung, total cell number of leukocyte and eosinophil in BALF, both EAR- I and EAR- III treated groups were significantly decreased compared to control group. IL-4, IL-5, IL-13, histamine, IgE in BLAF of group treated with both EAR-I and EM-III were significantly decreased compared to control group. In FACS analyzing, granulocytes, $CD3e^-$/$CCR3^+$, $CD3^+$/$CD19^$, $CD3e^+$/$CD69^+$, $CD4^+$, $CD8^+$ and $GR-1^+$/$CD11b^+$ were significantly increased in asthma induced mice group by OVA treatment compared to control group, and decreased in group treated with both EAR- I and EAR-III. Conclusions : The present data proves that extract of astragali radix has an effect on the inhibiting asthma. Moreover, astragali radix cultured for three years was proved to be superior to the one cultured for one year.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.11
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• pp.1625-1633
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• 2003

This study was conducted to observe effects of dietary acidogenicity value (AV) on rumen fermentation characteristics and nutrients digestibility. The AV of feedstuffs was based on the dissolution of Ca from $CaCO_3$ powder added at the end of a 24 h in vitro fermentation. Three diets were formulated to be iso-energetic and iso-nitrogenous with different AV. Two experiments were involved in this study. In experiment 1, it appears that pH, $NH_3-N$ concentration and A:P ratio tended to decrease, but gas production, VFA production and DM disappearance tended to increase with increasing dietary AV. In experiment 2, the rumen pH tended to decrease in order of high AV>medium AV>low AV treatment, respectively. There were no significant effects of dietary AV on $NH_3-N$ concentration, enzyme activity and nutrient digestibility. In addition, total VFA and individual VFA concentrations tended to increase with increasing dietary AV without significance. In fact, we hypothesized that different dietary AV would affect rumen fermentation and nutrients digestibility because dietary AV was adjusted with fermentable carbohydrate sources. The present results indicate that differences in dietary AV between treatments were too small to affect rumen fermentation and its effects were minimal.

• Journal of Periodontal and Implant Science
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• v.29 no.1
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• pp.31-40
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• 1999

Many factors may affect periodontal changes during the physiologic conditions of woman(e.g. puberty, menstrual cycle, pregnancy, menopause). Recently many research has focused on the immunological changes of host, but the exact mechanism is not clear. Collagen is a major constituent of periodontium, and collagenase specifically digests the collagen and plays a role in destruction of periodontal tissue. So, I suppose that it participates with the cytokines in the inflammation of gingiva and vascular response during the changes of female sex hormones. Because there are some evidences of the existence of the receptors of estrogen and progesterone in the gingiva, it may be a target tissue of female sex hormones. In this experiment, gingival fibroblast and periodontal ligament cell were cultured in the presence of various concentrations of estrogen or progesterone corresponding to the menstrual cycle and pregnancy. Collagenase activity of the supernatant of culture media was determined by Spectrophotometric collagenase assay. The enzyme activity was calculated by the % decrease of the coated collagen. 1. The estrogen at both concentrations had no effect on the activity of collagenase of the gingival fibroblast. 2. The progesterone had some effect on the collagenase activity of the gingival fibroblast at low and high concentration of menstrual cycle, and elevated the enzyme activity at all range of pregnancy concentrations. 3. In periodontal ligament cells, estrogen elevated the enzyme activity at the early pregnancy concentration and progesterone elevated at the concentration just before menstruation. In this experiment, pregesterone elevated the collagenase activity of gingival fibroblast and periodontal ligament cells. But the mechanism of the up-regulation of the enzyme activity was not confirmed. The more experiments of direct effect of progesterone on gingival at the molecular level(e.g. northern blot analysis) can reveal the exact mechanism.

• Journal of Gastric Cancer
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• v.20 no.1
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• pp.95-105
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• 2020

Purpose: Gastric cancer is a highly metastatic malignant tumor, often characterized by chemoresistance and high mortality. In the present study, we aimed to investigate the role of B-cell lymphoma 3 (Bcl-3) protein on cell migration and chemosensitivity of gastric cancer. Materials and Methods: The gastric cancer cell lines, AGS and NCI-N87, were used for the in vitro studies and the in vivo studies were performed using BALB/c nude mice. Western blotting, wound healing assay, Cell Counting Kit-8 assay, immunohistochemistry, and terminal deoxynucleotidyl transferase dUTP nick end labeling assay were used to evaluate the role of Bcl-3 in gastric cancer. Results: We found that the protein expression of hypoxia (HYP)-inducible factor-1α and Bcl-3 were markedly upregulated under hypoxic conditions in both AGS and NCI-N87 cells in a time-dependent manner. Interestingly, small interfering RNA-mediated knockdown of Bcl-3 expression affected the migration and chemosensitivity of the gastric cancer cells. AGS and NCI-N87 cells transfected with si-RNA-Bcl-3 (si-Bcl-3) showed significantly reduced migratory ability and increased chemosensitivity to oxaliplatin, 5-fluorouracil, and irinotecan. In addition, si-Bcl-3 restored the autophagy induced by HYP. Further, the protective role of si-Bcl-3 on the gastric cancer cells could be reversed by the autophagy inducer, rapamycin. Importantly, the in vivo xenograft tumor experiments showed similar results. Conclusions: Our present study reveals that Bcl-3 knockdown inhibits cell migration and chemoresistance of gastric cancer cells through restoring HYP-induced autophagy.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.4
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• pp.1823-1828
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• 2016

Background: Biological and pharmacological activities of dryocrassin ABBA, a phloroglucinol derivative extracted from Dryopteris crassirhizoma, have attracted attention. In this study, the apoptotic effect of dryocrassin ABBA on human hepatocellular carcinoma HepG2 cells was investigated. Materials and Methods: We tested the effects of dryocrassin ABBA on HepG2 in vitro by MTT, flow cytometry, real-time PCR, and Western blotting. KM male mice were used to detect the effect of dryocrassin ABBA on H22 cells in vivo. Results: Dryocrassin ABBA inhibited the growth of HepG2 cells in a concentration-dependent manner. After treatment with 25, 50, and $75{\mu}g/mL$ dryocrassin ABBA, the cell viability was 68%, 60% and 49%, respectively. Dryocrassin ABBA was able to induce apoptosis, measured by propidium iodide (PI)/annexin V-FITC double staining. The results of real-time PCR and Western ting showed that dryocrassin ABBA up-regulated p53 and Bax expression and inhibited Bcl-2 expression which led to an activation of caspase-3 and caspase-7 in the cytosol, and then induction of cell apoptosis. In vivo experiments also showed that dryocrassin ABBA treatment significantly suppressed tumor growth, without major side effects. Conclusions: Overall, these findings provide evidence that dryocrassin ABBA may induce apoptosis in human hepatocellular carcinoma cells through a caspase-mediated mitochondrial pathway.

• Journal of Nutrition and Health
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• v.36 no.2
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• pp.101-108
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• 2003

Antiplatelet aggregation, anticoagulant and lipid-lowering drugs are clinically widely used for secondary preventive purpose in the cardiovascular patients, but there is no primary preventive agents to prevent these diseases. With the aim of developing effective primary agents for cardiovascular diseases, we tried to formulate an optimized mixture of natural plants extract containing Theae sinensis, Camelliae sinensis, Vitis vinifera, Gingko folium and curcuminoids from Curcuma longa and to evaluate its anti-thrombotic and anti-hypercholesterolemic effects in vivo. The inhibitory effect of curcuminoids on vascular smooth muscle cell proliferation and migration were also investigated in vitro. in the animal experiments treated with hyperlipidemic diet, oral treatment of curcuminoids and natural plants extracts mixture (100 mg/kg) into male Sprague Dawley rats for 7 week simultaneously inhibited platelet aggregation as well as improved lipid profile in the blood. Compared to control group, both of curcuminoids-treated and mixture-treated groups revealed significantly decrease of total cholesterol (24.4%, 28.6%), free cholesterol (25.1%, 24.0%), cholesterol ester (14.6%, 29.0%), LDL-cholesterol (27.0%, 32.0%) and triglyceride (15.0%, 31.0%), respectively. However, both groups showed increase of HDL-cholesterol (46.6% and 51.5%) . In particular, atherogenic index of curcuminoids and mixture treatment group was significantly decreased to 47.0% and 56.0%, respectively. Furthermore, oral treatment of curcuminoids and mixture significantly inhibited collagen-induced platelet aggregation (21.1% and 29.1%, respectively), compared to control group. The anti-thrombotic values of mixture was almost similar to that of aspirin treatment (100 mg/kg) group. These results suggest that the oral treatment of curcuminoids-based natural plant extract mixture improved cardiovascular conditions in hyperlipidemic rats.

• Archives of Plastic Surgery
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• v.32 no.6
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• pp.689-698
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• 2005

Many studies for verifying angiogenesis have been in progress, especially in the field of abnormal vascular proliferation to explain the pathogenesis and to develop a treatment of several diseases. In our previous experiments, endothelial cell proliferations were induced by DMH stimulation in vitro, and the 177 factors(142 up-regulated and 35 down-regulated factors) were identified. Among the up-regulated factors, 9 substances (EFEMP1, CTGF, CYR61, $ITG{\beta}1$, FHL2, SERPINE1, MYC, PTTG1 and MSH6) were selected, which were related to cell proliferation and showed high signal intensities. The RNA was isolated from HUVECs at the time of 0, 6, 12, 24 hours after the DMH treatment, and RNA of control group HUVECs was also isolated. Genetic information of selected molecules was used to make primer for each, and RT-PCR was performed to analyze both groups. In control and treatment groups, each substance presented variety of manifestation degree according to time differences. EFEMP1, CTGF, CYR61, $ITG{\beta}1$, FHL2 and MYC were related to abnormal vascular proliferation steadily and SERPINE1, PTTG1 and MSH6 were related secondarily. CTGF was related to both normal and abnormal proliferation, but it played a more significant role in abnormal proliferation from earlier stage. EFEMP1, CYR61, $ITG{\beta}1$, FHL2 and MYC were similar to CTGF, although the relation appeared lately. Further study should be performed to analyze the expressions and the interactions of growth factors, which could be utilized in the new therapeutic development.

• Imaging Science in Dentistry
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• v.38 no.4
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• pp.195-202
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• 2008

Purpose : To characterize the effects of 2-deoxy-D-glucose (2-DG) and quercetin (QCT) on gene expression of osteonectin (ON) and osteopontin (OP) in irradiated MC3T3-El cells. Materials and Methods : When MC3T3-El osteoblastic cells had reached 70-80% confluence, cultures were transferred to a differentiating medium supplemented with 5 mM 2-DG or 10 ${\mu}M$ QCT and then irradiated with 2, 4, 6, and 8 Gy. At various times after irradiation, the cells were analyzed for the expression of bone mineralization genes such as ON and OP. Results : The mRNA expression of both ON and OP was increased according to the culture time in the differentiation medium, and the increase of the genes peaked at 14 days after the differentiation induction. In the case of OP, the increase of mRNA expression was maintained to 28 days after the differentiation, while the mRNA level of ON was reduced to the basal level at the same time. Irradiation adding 2-DG showed a significant peak value in the expression pattern of ON at 4 Gy 7 days after irradiation. Irradiation adding QCT increased the mRNA expression of ON and OP in a dose-dependant manner, but irradiation adding 2-DG did not show any differences between the control and experiments 14 days after irradiation. Irradiation adding QCT increased significantly the expression patterns of ON 21 days after irradiation. Conclusion : The results showed that QCT acted as a radiosensitizer in the gene expression of ON and OP during differentiation of the late stage of irradiated MC3T3-E1 osteoblastic cells in vitro. (Korean J Oral Maxillofac Radiol 2008; 38: 195-202)