• Clinical and Experimental Reproductive Medicine
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• v.28 no.3
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• pp.191-198
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• 2001

Objective : This study was conducted to examine the effect of vitrification on the survival and in vitro development of mice 1-cell zygotes. Method: Effects of exposure to vitrification solution and vitrification, with different concentrations of the cryoprotectant solution, were examined. The 1-cell zygotes were also subjected to a slow freezing-thawing method to compare with vitrification method. Solution composed of ethylene glycol (6.0 M, 5.0 M, 4.0 M) and sucrose (1.0 M) were used as cryopropectant. The experiments employed the method loading the embryos on electron microscope grids. Results: I. The effects of exposure in vitrification solution. 1-cell zygotes were non-toxic at all concentrations of the vitrification solution showing the survival rate between 88.1% and 97.5%. Development into 2-cell was more successful in the higher concentrations of the vitrification solution. Therefore, higher concentrations of the vitirification solution do not seem to cause any problems in vitrification procedure. II. The effects of vitrification method. 1-cell zygotes showed the survival rate between 78.8% and 92.4%. The lowest and the highest survival rate was observed in the 6.0 M and 4.0 M vitrification solution, respectively. 2-cell development rates varied from 77.6% to 91.3%. Blastocyst development rate was shown highest in 5.0 M and the lowest in 4.0 M solution. Therefore, the highest 2-cell and blastocyst development rate was observed in 5.0 M solution. III. Comparison of vitrification and slow freezing-thawing method on 1-cell zygotes. This experiment showed that 1-cell zygotes had the highest survival and development rates in 5.0 M vitrification solution. Vitrified group of 1-cell zygotes, in the 5.0 M vitrification solution, were compared with the group processed in slow freezing-thawing method. The development rate into 2-cell and blastocyst as well as the survival rate were higher in the vitrified group than in the slowly freezed group. Conclusion: 1. The results demonstrate that the best cryoprotectant is a 5.0 M vitrification solution for 1-cell zygotes. 2. Vitrification method significantly increases the survival rate of the 1-cell zygote and its development into 2-cell and blastocyst. Equilibration and exposure time during the vitrification was remarkerbly short in this experiment. Total time, from the exposure to vitirification solution to storage in the liquid nitrogen, was taken only 90 seconds. In contrast, the slow freezing-thawing method have taken more than four hours. Taken together, we presume that the overall time used for the procedure contributes to the results as an important parameter. 3. The loading of 1-cell zygotes on the EM grid is technically more simple and takes less time than the straw or cryo vial method.

• Journal of Physiology & Pathology in Korean Medicine
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• v.27 no.1
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• pp.26-33
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• 2013

The present study explored studies directed at elucidating the effectiveness of Korean medicine in the treatment of sterility and systematically analyzed the trends of studies on sterility. The standardization and objectification based on results for this study may contribute to the development of effective Korean medicine in reducing sterility. In the annual tendency of sterility-related clinical studies, the number of papers began to increase rapidly from 2000, and 15 papers were published from 2000 till 2011. Then, in the technical journals publishing them, The Journal of Oriental Obstetrics & Gynecology published 13 the most papers. In the institutions to which the first authors belonged, Dongguk University Hospital had 5 the most papers. In the number of test subjects was also analyzed and as the result the studies with 1~9 were the most as 10. In the study design analysis, there were 10 case reports, 5 case series and 4 before and after studies. In the analysis of intervention methods, there were 10 papers where only the Korean medicines were given, next 6 papers where the acupuncture and moxibustion were added, and 3 papers where the Korean medical therapies and other subsidiary therapies were used. In the analysis on the existence of pattern identification, there were 12 papers with the pattern identification, 6 without the pattern identification and 1 with the four constitution identification. In the IRB approval, 18 or most papers had not received the IRB approval and just 1 paper got the IRB approval. When the sterility-related clinical studies published in the Korean medical journals were surveyed, it turned out that there are no RCT studies and there is just one study with the IRB approval. And there were 6 papers without the pattern identification of Korean medicine. So, it seems necessary in the future to improve the papers in these respects for the development of Korean medicine sterility treatment with a good basis through the more systematic and accurate study method.

• Journal of Embryo Transfer
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• v.31 no.4
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• pp.367-374
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• 2016

This study was conducted to evaluate the effect of transfer temperature of epididymis on survival rate of semen and development ability of B6D2F1 mice embryos. No significant differences were noted in the survival rate of semen ($59.0%{\pm}0.1$ vs. $47.6%{\pm}0.1$), in vitro fertilization rate ($90.7%{\pm}0.1$ vs. $90.7%{\pm}0.1$), developmental rate ($90.0%{\pm}0.1$ vs. $90.0%{\pm}0.1$), and blastocysts formation rate ($53.1%{\pm}0.2$ vs. $52.3%{\pm}0.2$) between groups. (NS; P>0.05). However, the zona hatched rate was significantly higher in the $4^{\circ}C$ group compared to those of the $37^{\circ}C$ group ($47.8%{\pm}0.1$ vs. $25.6%{\pm}0.2$; p<0.05). When it comes to cell numbers of blastocysts, the % ICM (/total cells) was significantly higher in the group of $4^{\circ}C$ compared to the $37^{\circ}C$ ($27.0%{\pm}0.1$ vs. $18.3%{\pm}0.1$; p<0.05). However there were no differences in total cell numbers ($72.7{\pm}31.6$ vs. $62.0{\pm}36.6$), ICM cell numbers ($17.0{\pm}7.8$ vs. $14.6{\pm}8.6$), TE cell numbers ($55.8{\pm}29.8$ vs. $64.0{\pm}24.4$), the ratio of ICM:TE ($1:4.2{\pm}4.1$ vs. $1:6.4{\pm}7.2$) between two groups (NS; P>0.05). Taken altogether, it is expected to achieve the best developmental ability of B6D2F1 mice embryos in the transfer temperature of epididymis. Also these results can provide fundamental data to maximize culture condition for in vitro fertilization on B6D2F1 mice. In future, therefore, it is expected that results herein might be applied for in vitro culture of human embryos.

• Journal of Nutrition and Health
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• v.39 no.5
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• pp.467-475
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• 2006

The necessity of adequate pregnancy weight gain for optimal pregnancy outcome has been recognized. However, the specific components of pregnancy weight gain that might be critical for fetal growth and development have not been elucidated clearly. The purpose of this study was to investigate the correlation of pregnancy weight gain and birth weight with serum leptin levels in women delivered newborns. The subjects were recruited from K university hospital. The subject's characteristic data (age $32.1\;{\pm}\;4.3\;y$, gestational age $39.5\;{\pm}\;1.1wk$, pre-pregnancy weight $58.0\;{\pm}\;8.6\;kg$, pregnancy weight gain $12.7\;{\pm}\;5.5\;kg$, newborn's birth weight $3.5\;{\pm}\;0.5\;kg$) were gathered. Maternal dietary assessment was carried out at the end of pregnancy. After delivery, blood samples were collected from 20 mother-newborn pairs. Serum levels of various lipids and leptin were analyzed. Maternal daily consumption of iron, zinc, folate were lower than the RDA of each nutrient and index of nutritional quality was less than 1 showing that the quality of maternal diet was low. The levels of serum leptin of mothers and infants were $10.2\;{\pm}\;6.7\;ng/ml$ and $1.7\;{\pm}\;0.6\;ng/ml$, respectively. The serum leptin concentrations of male infants $(1.9\;{\pm}\;0.7\;ng\;ml)$ were not different from that of females $(1.7{\pm}0.5\;ng/ml)$. A negative correlation was found between the maternal pre-pregnant BMI and weight gain during pregnancy (r = -0.54, p < 0.05). There was a positive correlation between the pregnancy weight gain and the newborn's birth weight (r=0.59, p < 0.01 There were also positive correlation between newborn's birth weight and newborn's serum leptin levels (r = 0.57, p < 0.01). No correlations were found between maternal serum leptin levels and that of newborn's. Efforts should be made to attain adequate diet and weight gain during the pregnancy to reduce the likelihood of low or over birth weight of newborns.

• Clinical and Experimental Reproductive Medicine
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• v.46 no.4
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• pp.173-177
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• 2019

Objective: We investigated the clinical characteristics of men with testosterone replacement therapy (TRT)-induced hypogonadism and its effect on assisted reproductive technology (ART) in infertile couples. Methods: This study examined the records of 20 consecutive male patients diagnosed with azoospermia or severe oligozoospermia (< 5 × 10⁶/mL) who visited a single infertility center from January 2008 to July 2018. All patients were treated at a primary clinic for erectile dysfunction or androgen deficiency symptoms combined with low serum testosterone. All men received a phosphodiesterase 5 inhibitor and TRT with testosterone undecanoate (Nebido^®) or testosterone enanthate (Jenasteron^®). Patients older than 50 years or with a chronic medical disease such as diabetes were excluded. Results: The mean age of patients was 37 years and the mean duration of infertility was 16.3 ± 11.6 months. At the initial presentation, eight patients had azoospermia, nine had cryptozoospermia, and three had severe oligozoospermia. Serum follicle-stimulating hormone levels were below 1.0 mIU/mL in most patients. Three ongoing ART programs with female factor infertility were cancelled due to male spermatogenic dysfunction; two of these men had normal semen parameters in the previous cycle. After withholding TRT, serum hormone levels and sperm concentrations returned to normal range after a median duration of 8 months. Conclusion: TRT with high-dose testosterone can cause spermatogenic dysfunction due to suppression of the hypothalamic-pituitary-testicular axis, with adverse effects on infertility treatment programs. TRT is therefore contraindicated for infertile couples attempting to conceive, and the patient's desire for fertility must be considered before initiation of TRT in a hypogonadal man.

• Clinical and Experimental Reproductive Medicine
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• v.36 no.2
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• pp.101-109
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• 2009

Objective: This study was carried out to analyze the amount of telomeric DNA and telomerase activity in early bovine embryos. Methods: The amount of telomeric DNA in early bovine embryos at the 8 cell, morula and blastocyst stages was analyzed by Quantitative Fluorescence In Situ Hybridization (Q-FISH) technique using a bovine telomeric DNA probe. Telomerase activity was analyzed by Telomeric Repeat Amplification Protocol (TRAP assay). Results: The relative amount of telomeric DNA in early bovine embryos was gradually increased from 8 cell to blastocyst stage. It was not significantly associated with the grade of embryo quality. While telomerase activity was detected in the early bovine embryos at these stages, it significantly increased at morula stage and showed maximum activity at the blastocyst stage. Conclusion: The amount of telomeric DNA and the telomerase activity of bovine embryos increase during the progression of early embryogenesis, suggesting a positive correlation between telomeric DNA and telomerase activity. The telomerase activity seems to increase to maintain the levels of telomeric DNA through embryo development which are required for extensive cell division.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.12
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• pp.6391-6395
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• 2012

Background: Ovarian cancer is ranked as the fifth most common cause of cancer death in women. In Malaysia, it is the fourth most common cancer in females. CA125 has been the tumor marker of choice in ovarian cancer but its diagnostic specificity in early stages is only 50%. Hence, there is a critical need to identify an alternative tumor marker that is capable of detecting detect ovarian cancer at an early stage. HE4 is a new tumor marker proposed for the early diagnosis of ovarian cancer and disease recurrence. Currently, none of the normal ranges of HE4 quoted in the literature are based on data for a multiethnic Asian population. Therefore, the aim of this study was to determine reference intervals for HE4 in an Asian population presenting in University Malaya Medical Centre, a tertiary reference hospital. Materials and Methods: 300 healthy women were recruited comprising 150 premenopausal and 150 postmenopausal women, aged from 20-76 years. All women were subjected to a pelvic ultrasonograph and were confirmed to be free from ovarian pathology on recruitment. Serum HE4 levels were determined by chemiluminescent microparticle immunoassay (CMIA, Abbott Architect). The reference intervals were determined following CLSI guidelines (C28-A2) using a non-parametric method. Results: The upper limits of the $95^{th}$ percentile reference interval (90%CI) for all the women collectively were 64.6 pmol/L, and 58.4 pmol/L for premenopausal) and 69.0 pmol/L for postmenopausal. The concentration of HE4 was noted to increase with age especially in women who were more than 50 years old. We also noted that our proposed reference limit was lower compared to the level given by manufacturer Abbott Architect HE4 kit insert (58.4 vs 70 pmol/L for premenopausal group and 69.0 vs 140 pmol/L in the postmenopausal group). The study also showed a significant difference in HE4 concentrations between ethnic groups (Malays and Indians). The levels of HE4 in Indians appeared higher than in Malays (p<0.05), while no significant differences were noted between the Malays and Chinese ethnic groups. Conclusions: More data are needed to establish a reference interval that will better represent the multiethnic Malaysian population. Probably a larger sampling size of equal representation of the Malay, Chinese, Indians as well as the other native ethnic communities will give us a greater confidence on whether genetics plays a role in reference interval determination.

• Radiation Oncology Journal
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• v.17 no.2
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• pp.120-129
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• 1999

Purpose : Serum squamous cell (SCC) antigen levels were examined in uterine cervix cancer undergoing radiation therapy, and authors analyzed the relationship between SCC antigen levels and treatment results. Materials and Methods :This is a retrospective study of 181 conical carcinoma patients who received radiotherapy and examined serial serum SCC antigen from 1991 to 1997 at Soonchunhyang University Hospital. One hundred and eighteen patients underwent SCC antigen evaluation at diagnosis The relationship between the serum tumor marker level and disease free survival, recurrence pattern, and other prognostic factors were analyzed according to various statistical methods. Results : The Positivity rate (initial serum value above 2.5 ng/ml) was increased with FIGO stage (IB-IIA 57% to IV 91%) and more discriminative than cutoff value of 1.5 ng/ml. Five year disease free survival rates for the stage IB-IIA, IIB, III and IV were 79.2%, 68.7%, 33.4% and 0%, respectively. The 5-year disease free survival rate for patients with serum SCC antigen levels above 5.0 ng/ml was 34% versus 55~62% for patients with normal range (>1.5 ng/ml) or mildly elevated levels (1.5~5.0 ng/ml). Rising SCC antigen levels preceded the clinical detection of disease by a mean of 4.8 months (range 1 ~13 months). Negative linear correlation was observed between initial SCC antigen levels and relapse free survival (r=-0.226), and by multivariate analysis, initial SCC antigen level had a large impact on the relapse free survival. Conclusion : SCC antigen assay is a useful aid to predict the prognosis of squamous cell carcinoma of the uterine cervix and to detect recurrence.

• Parasites, Hosts and Diseases
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• v.35 no.3
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• pp.189-196
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• 1997

The purpose of this study is to determine whether nitric oxide is involved in the extracellular killing of Trichomoncs uasinalis by mouse (BALB/c) peritoneal macrophages and RAW264.7 cells activated with LPS or rIFN-γ and also to observe the effects of various chemicals which affect the production of reactive nitrogen intermediates (RNl) in the cytotoxicity against T. vnginnlis. The cytotoxicity was measured by counting the release of (3H)-thymidine from labelled protozoa and NOa was assayed by Griess reaction. Nemonomethyl-L-arginine (L-NMHA), Nenitro-L-arginine methyl ester (NAME) and arginase inhibited cytotoxicity to T. vaginnlis and nitrite production by activated mouse perioneal macrophagrs and RAW 264.7 cells. The addition of excess L-arginine competitively restored trichomonacidal activity of macrophages. Exogenous addition of FeSO4 inhibited cytotoxicity to T. vaginaLis and nitric products of macrophages. From above results, it is assumed that nitric oxide plays an important role in the host defense mechanism of macrophages against T ucfinalis.

• Parasites, Hosts and Diseases
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• v.23 no.2
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• pp.293-299
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• 1985

This study is to verify the protective ability against experimental Naegleria meningoencephalitis by immunization with Naegleria fowleri in mice. Naegleria fewleri, strain 0359, and Naegleria gruberi, strain EGB, were used in this study, and cultured in CGVS medium akenically. Inbred BALB/C mice, weighing about 20g, were immunized by three intraperitoneal injection of $1{\times}10^6$ N. fowleri trophozoites at the interval of one week. This N. fowleri trophozoites antigen was fixed with 5% formaldehyde. N. fowleri trophozoites from culture were homogenized with soiicator at $4^{\circ}C$ as monitored by phase contrast microscopy, and their membrane and cell content preparations were made for the immunization of mice. Their inoculation dose in volume was equivalent to the $1{\times}10^6$ trophozoites in each injection for immunization. And N. gruberi trophosoites, whieh was fixed with 5% formaldehyde, were also used for immunisation. Mice were inoculated intranasally with $5{\times}10^4$ N. fowleri trophozoites in a 511 suspension under anesthesia by as intraperitoneal injection of about 1 mg secobarbiturate. Nervousness, rotation or sluggish behaviour were observed in the mice which were infected with N. fewleri. Necrotic lesion was demonstrated in the anterior portion of brain, especially in the olfactory lobe. The inflammatory cell infiltration with numerous H. fowleri trophozoites was noticed. This pathological changes were more extensive in the control than in the experimental groups. Mice were dead due to experimental primary amoebic meningoencephalitis that developed between 8 days and 23 days after inoculation. Mortality rate of the mice was low in the immunized experimental group. Mean survival time, which is the survival duration of mice from the infection to death, was prolonged significantly in the immunized mice except in the mice immunized with JV, fowleri membrane. Even in the mice immunized with N. gruberi, survival time was delayed. In summary, the effectiveness of immunization is demonstrated in terms of protective immunity against Naegleria meningoencephalitis in mice.