• Clinical and Experimental Reproductive Medicine
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• v.45 no.3
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• pp.129-134
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• 2018

Objective: In frozen and thawed embryos, the zona pellucida (ZP) can be damaged due to hardening. Laser-assisted hatching (LAH) of embryos can increase the pregnancy rate. This study compared thinning and drilling of the ZP before frozen embryo transfer (FET). Methods: Patients were randomly allocated into two groups for LAH using thinning or drilling on day 2 after thawing. Twenty-five percent of the ZP circumference and 50% of the ZP thickness was removed in the thinning group, and a hole $40{\mu}m$ in diameter was made in the drilling group. Results: A total of 171 in vitro fertilization/intracytoplasmic sperm injection FET cycles, including 85 cycles with drilling LAH and 86 cycles with thinning LAH, were carried out. The thinning group had a similar ${\beta}$-human chorionic gonadotropin-positive rate (38.4% vs. 29.4%), implantation rate (16.5% vs. 14.4%), clinical pregnancy rate (36.0% vs. 25.9%), miscarriage rate (5.8% vs. 2.4%), ongoing pregnancy rate (30.2% vs. 23.5%), and multiple pregnancy rate (7.0% vs. 10.6%) to the drilling LAH group. There were no significant differences in pregnancy outcomes between subgroups defined based on age (older or younger than 35 years) or ZP thickness (greater or less than $17{\mu}m$) according to the LAH method. Conclusion: The present study demonstrated that partial ZP thinning or drilling resulted in similar outcomes in implantation and pregnancy rates using thawed embryos, irrespective of women's age or ZP thickness.

• Journal of Yeungnam Medical Science
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• v.15 no.2
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• pp.286-296
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• 1998

Different techniques for culturing respiratory epithelial cells have been developed to overcome the limitations of studies on in vivo and on bioptic material. Traditionally, culture systems are divided into organ cultures, explant cultures and dissociated cell cultures. The first two contain both epithelial and non-epithelial cells. However, in monolayer cultures of dissociated cells only epithelial cells are present, the effects observed are caused by a pure epithelial responses. The purpose of this study is to establish primary culture method of human nasal epithelium (HNEC) by monolayer culture of dissociated cells to evaluate the role of the epithelial cells in the allergic and non-allergic nasal inflammatory reactions. HNEC was prepared by primary culture method of monolayer culture of dissociated cells from human inferior nasal turbinate mucosa of septal deviation patients. Primary cultured cells were characterized by indirect immunofluorescence assay and transmission electron microscopy. The immunoreactivities of cytokeratin-pan and cytokeratin No. 8 were observed in cultured HNEC. However, the immnoreactivities of vimentin and von Willebrand factor were not observed in cultured HNEC. The tonofilaments and desmosome were observed in cultured HNEC. The cultured epithelial cells were identified to be pure nasal epithelial cells. The monolayer culture of dissociated cells could successfully be employed for further study to investigate the role of the epithelial cells in allergic or non-allergic nasal inflammatory diseases.

• 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.

• Archives of Plastic Surgery
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• v.37 no.5
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• pp.531-534
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• 2010

Purpose: Lipobean$^{(R)}$s, widely used in lipodissolving techniques, contain phosphatidylcholine and sodium deoxycholate as its main substances. They have been approved only as medication for liver disease by the FDA. However, they have been used under various clinical settings without exact knowledge of its action mechanism. The authors designed an in vitro study to analyze the effects of different concentrations of phosphatidylcholine and sodium deoxycholate on adipocytes and other types of cells. Methods: Human adipose-derived stem cell were cultured and induced to differentiate into adipocytes. Fibroblasts extracted from human inferior turbinate tissue, and MC3T3-E1 osteoblast lines were cultured. Phosphatidylcholine solution dissolved with ethanol was applied to the culture medium at differing concentrations (1, 4, 7, 10 mg/mL). The sodium deoxycholate solution dissolved in DMSO applied to the medium at differing concentrations (0.07, 0.1. 0.4. 0.7 mg/mL). Cells were dispersed at a concentration of $5{\times}10^3$ cells/well in 24 well plates, and surviving cells were calculated 1 day after the application using a CCK-8 kit. Results: The number of surviving cells of adipocytes, fibroblasts and osteoblasts decreased as the concentration of sodium deoxycholate increased. However, all types of cells that had been processed in a phosphatidylcholine showed a cell survival rate of over 70% at all concentrations. Conclusion: This study shows that sodium deoxycholate is the more major factor in destroying adipocytes, and it is also toxic to the other cells. Therefore, we conclude that care must be taken when using Lipobean$^{(R)}$s as a method of reducing adipose tissue, for its toxicity may destroy other nontarget cells existing in the subcutaneous tissue layer.

• Journal of Periodontal and Implant Science
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• v.34 no.1
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• pp.49-59
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• 2004

The goal of periodontal treatment is not only to arrest the progression of the disease but also to promote the functional, esthetic regeneration of the periodontium. Flap operation, bone graft, guided tissue regeneration, growth factors and bone morphogenetic protein have been used for this purpose. Among these techniques of regeneration, alloplastic graft, especially calcium phosphate is getting more attention recently. The purpose of this study was to evaluate the effects of calcium phosphate glass on mouse calvarial cell in vitro. The toxicity of calcium phosphate glass was measured using MTT assay, the synthesis of collagen was measured using collagen assay, and ALP activity was measured. The experimental groups were cultured with calcium phosphate glass(both AQ-, and HT-CPG) in concentration of 0.01, 0.02, 0.1, 0.2g/ml. The results are as follows 1. In concentrations not exceeding 0.02g/ml, both the groups(AQ-CPG, HT-CPG) didn't show any toxicity on mouse calvarial cell(p<0.05). 2. In both the experimental groups are the concentration of 0.02g/ml, collagen expressions were significantly up-regulated (p<0.05). 3. In both the experimental groups are the concentration of 0.02g/ml, ALP activity was not significantly up-regulated, but ALP activity in both experimental groups were greater than control group(p<0.05). The results suggested that the use of calcium phosphate glass may promotes periodontal regeneration. Ongoing studies are necessary in order to determine their regeneration effects.

• Journal of Korean Society of Forest Science
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• v.99 no.1
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• pp.125-130
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• 2010

The application of bioreactor culture techniques for plant micropropagation is regarded as one of the ways to reduce production cost by scaling-up and automation. In an attempt to optimize mass proliferation systems in Eucalyptus pellita, four types of bioreator systems including temporary immersion system with or without net were tested. Highest growth was achieved with 30-min flushes of medium at every 4-h intervals in TIN (temporary immersion with net) system. Results indicate over three-fold increase in shoot growth with the TIN system when compared with TIX (control: temporary immersion without net) system which is without net in bioreactor. Furthermore, plants produced from the TIN system increased total chlorophyll content, chlorophyll a/b and dry matter, giving higher yields of acclimatized plants. Our findings suggest that plantlet growth increases with appropriate exposure to media at correct intervals, as well as use of net for maintaining aerobic condition in the vessels. The TIN system thus has great potential for in vitro mass production of Eucalyptus clones commercially.

• Korean Journal of Medicinal Crop Science
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• v.3 no.1
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• pp.40-44
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• 1995

This study was aimed to mutant induction in Gentiana scabra BUNGE through the tissue culture techniques and comparison of gentiopicroside content in each mutant plants derived from tissue culture. In order to induce the mutants of Gentiam scabra, seed, apical and lateral bud were soaked in $NaN_3$, EMS and MNU solutions and cultured on MS basal medium containing 0.5mg/l NAA, BA $NaN_3$, MNU-treated buds survived about 50% but not survived in EMS treatment. Seed germination rate was extremely low in EMS and MNU treatments but various types of mutant plantlets were obtained by those treatments. 63% of elongated type and 37% of dwarf type were obtained from tissue culture treated by various mutagen. Gentiopicroside contents of regenerated plantlets was analyzed. Root of dwarf type contained more gentiopicroside(1.383%) than that of elongation type (0.083%).

• Journal of Periodontal and Implant Science
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• v.50 no.1
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• pp.56-66
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• 2020

Purpose: A stability-measuring device that utilizes damping capacity analysis (DCA) has recently been introduced in the field of dental implantology. This study aimed to evaluate the sensitivity and reliability of this device by measuring the implant stability of ex vivo samples in comparison with a resonance frequency analysis (RFA) device. Methods: Six implant beds were prepared in porcine ribs using 3 different drilling protocols to simulate various implant stability conditions. Thirty-six pork ribs and 216 bone-level implants measuring 10 mm in height were used. The implant beds were prepared using 1 of the following 3 drilling protocols: 10-mm drilling depth with a 3.5-mm-diameter twist drill, 5-mm drilling depth with a 4.0-mm-diameter twist drill, and 10-mm drilling depth with a 4.0-mm-diameter twist drill. The first 108 implants were external-connection implants 4.0 mm in diameter, while the other 108 implants were internal-connection implants 4.3 mm in diameter. The peak insertion torque (PIT) during implant placement, the stability values obtained with DCA and RFA devices after implant placement, and the peak removal torque (PRT) during implant removal were measured. Results: The intraclass correlation coefficients (ICCs) of the implant stability quotient (ISQ) results obtained using the RFA device at the medial, distal, ventral, and dorsal points were 0.997, 0.994, 0.994, and 0.998, respectively. The ICCs of the implant stability test (IST) results obtained using the DCA device at the corresponding locations were 0.972, 0.975, 0.974, and 0.976, respectively. Logarithmic relationships between PIT and IST, PIT and ISQ, PRT and IST, and PRT and ISQ were observed. The mean absolute difference between the ISQ and IST values on a Bland-Altman plot was -6.76 (-25.05 to 11.53, P<0.05). Conclusions: Within the limits of ex vivo studies, measurements made using the RFA and DCA devices were found to be correlated under a variety of stability conditions.

• The Journal of the Convergence on Culture Technology
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• v.5 no.1
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• pp.413-416
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• 2019

Alstroemeria (Alstroemeriaceae) is one of the most important cut flowers in international market. Especially, characteristics like long vase-life, various colors, tolerance to low temperature and a low energy requirement during cultivation have stimulated this success. Because of its characteristics such as low multiplication rates, time-consuming process and high risk of carrying viral disease, in vitro propagation techniques based on rhizome meristems culture have been developing nowadays. The callus induction has various cultivation sites compared with the direct plant generation method, and if the callus is maintained well, the plant differentiation can be performed simultaneously while maintaining the callus, so that it can be used for mass proliferation. In this study, we tested various hormones and cultivars for efficient callus induction. As a result of culturing between the nodes and the internodes, the callus began to be formed after 8 weeks, and the calli incidence in the nodes was higher than that between the internodes. Also, in the comparison of 2,4-D and picloram, the callus incidence rate was up to 2 times higher in the medium treated with 2,4-D. Using these results, it is thought that it will help establish the system of mass propagation system of Alstroemeria and cultivate new varieties.

• Journal of Ginseng Research
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• v.45 no.1
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• pp.48-57
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• 2021

Background: Ginsenoside Rh2 is well known for many pharmacological activities, such as anticancer, antidiabetes, antiinflammatory, and antiobesity properties. Glycosyltransferases (GTs) are ubiquitous enzymes present in nature and are widely used for the synthesis of oligosaccharides, polysaccharides, glycoconjugates, and novel derivatives. We aimed to synthesize new ginsenosides from Rh2 using the recombinant GT enzyme and investigate its cytotoxicity with diverse cell lines. Methods: We have used a GT gene with 1,224-bp gene sequence cloned from Lactobacillus rhamnosus (LRGT) and then expressed in Escherichia coli BL21 (DE3). The recombinant GT protein was purified and demonstrated to transform Rh2 into two novel ginsenosides, and they were characterized by nuclear magnetic resonance (NMR) techniques and evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2-5-diphenyltetrazolium bromide assay. Results: Two novel ginsenosides with an additional glucopyranosyl (6→1) and two additional glucopyranosyl (6→1) linked with the C-3 position of the substrate Rh2 were synthesized, respectively. Cell viability assay in the lung cancer (A549) cell line showed that glucosyl ginsenoside Rh2 inhibited cell viability more potently than ginsenoside Rg3 and Rh2 at a concentration of 10 μM. Furthermore, glucosyl ginsenoside Rh2 did not exhibit any cytotoxic effect in murine macrophage cells (RAW264.7), mouse embryo fibroblasts cells (3T3-L1), and skin cells (B16BL6) at a concentration of 10 μM compared with ginsenoside Rh2 and Rg3. Conclusion: This is the first report on the synthesis of two novel ginsenosides, namely, glucosyl ginsenoside Rh2 and diglucosyl ginsenoside Rh2 from Rh2 by using recombinant GT isolated from L. rhamnosus. Moreover, diglucosyl ginsenoside Rh2 might be a new candidate for treatment of inflammation, obesity, and skin whiting, and especially for anticancer.