• Journal of Embryo Transfer
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• v.31 no.3
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• pp.281-285
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• 2016

Cluster-of-differentiation antigen 9 (CD9) gene expressed in the male germ line stem cells is crucial for sperm-egg fusion, and was therefore selected as a candidate gene to investigate Duroc boar semen motility and kinematic characteristics. This study was performed to investigatetheir association with semen motility and kinematic characteristics. DNA samples from 96 Duroc pigs with records of sperm motility and kinematic characteristics [Total motile spermatozoa (MOT, $82.27{\pm}5.58$), Curvilinear velocity(VCL, $68.37{\pm}14.58$), Straight-line velocity(VSL, $29.06{\pm}6.58$), the ratio between VSL and VCL(LIN, $47.36{\pm}8.42$), Amplitude of Lateral Head displacement(ALH, $2.88{\pm}0.70$)] were used in present study. A single nucleotide polymorphism (g.358A>T) in intron 6 was associated with MOT, VCL, VAP and ALH in Duroc population (p<0.05). Therefore, we suggest that the porcine CD9 may be used as a molecular marker for Duroc boar semen quality, although its functional effect was not clear yet. These results will improve the understanding of the functions of the CD9 in spermatogenesis within the reproductive tracts, and will shed light on CD9 as a candidate gene in the selection of good sperm quality boars.

• Korean Journal of Pharmacognosy
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• v.40 no.2
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• pp.103-108
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• 2009

GCSB-5 preparation is a purified extract from a mixture of 6 medicinal plants(Acanthopanacis Cortex, Achyranthis Radix, Saposhnikoviae Radix, Cibotii Rhizoma, Glycine Semen Nigra, Eucommiae Cortex) that have been widely used for the treatment of various bone disorders. The aim of this study was to investigate HPLC analysis method and screening of standard compound on Saposhnikoviae Radix for quality standardization of a medicinal crude drug GCSB-5. Standard compound of Saposhnikoviae Radix was decided with cimifugin by isolation and instrumental analysis such as NMR. HPLC analysis method for the simultaneous determination of cimifugin was established for the quality control of the medicinal plants of Saposhnikoviae Radix species, GCSB-5 raw material and preparation. And validation of HPLC analysis methods were conformed for verification of HPLC methods by check to specificity, linearity, intra-day precision, inter-day precision and accuracy following ICH guideline.

• Journal of Embryo Transfer
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• v.30 no.1
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• pp.7-15
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• 2015

The objective of this study was to investigate the efficiency of nicotinic acid on sperm cryosurvival and fertilization ability in frozen-thawed boar semen. Boar semen was collected by glove-hand method and was frozen using freezing solution treated to 0, 5, 10 and 20 mM of nicotinic acid. The frozen sperm for sperm characteristic analysis was thawed such as viability, acrosome reaction, and mitochondrial integrity. The frozen-thawed sperm was estimated by SYBR14/PI double staining for viability, FITC-PNA/PI double staining for acrosome reaction and Rhodamine123/PI double staining for mitochondrial integrity using a flow cytometry. The embryo was estimated in vitro development and DCFDA staining for reactive oxygen species assessment. As results, frozen-thawed sperm viability was significantly higher in 5 and 10 mM ($61.1{\pm}1.5%$,$64.7{\pm}2.0%$) of nicotinic acid than other groups (0 mM, $52.1{\pm}2.3%$; 20 mM, $47.8{\pm}5.1%$, P<0.05). The live sperm with acrosome reaction was significantly higher in 5 and 10 mM of nicotinic acid ($26.1{\pm}1.8%$, $24.9{\pm}1.5%$) than other groups (0 mM, $35.3{\pm}0.8%$; 20 mM, $36.5{\pm}1.9%$, P<0.05). The live sperm with mitochondrial integrity was significantly higher in 5 and 10 mM ($84.2{\pm}3.6%$, $88.4{\pm}2.3%$) of nicotinic acid than other groups (0 mM, $77.3{\pm}4.4%$; 20 mM, $73.3{\pm}3.6%$, P<0.05). Blastocyst rate of in vitro development was significantly higher in 10 mM ($17.0{\pm}1.3%$) of nicotinic acid than other groups (0 mM, $9.4{\pm}0.5%$; 5mM, $12.6{\pm}0.8%$; 20 mM, $5.0{\pm}1.0%$, P<0.05). Moreover, total cell number was higher in 5 and 10 mM ($53.6{\pm}2.9%$, $57.9{\pm}2.8%$) of nicotinic acid than other groups (0 mM, $41.0{\pm}1.4%$; 20 mM, $23.2{\pm}2.8%$, P<0.05). Hydrogen peroxide in embryos was lower in 5 mM nicotinic acid ($0.7{\pm}0.1%$) than other groups (0 mM, $1.0{\pm}0.1%$; 10mM, $0.9{\pm}0.0%$; 20 mM, $1.4{\pm}1.0%$, P<0.05). In conclusion, nicotinic acid-treated semen improves cryosurvival and quality of spermatozoa. Also, the fertilized oocytes with nicotinic acid improve quality of embryo and blastocyst formation.

• ANNALS OF ANIMAL RESOURCE SCIENCES
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• v.29 no.4
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• pp.150-157
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• 2018

The Zygote arrest 1 (ZAR1) gene is known to affect early embryonic development in various vertebrates. In this study, we performed the association analysis to check whether there is any significant relationship between semen kinematic characteristics and the ZAR1 gene. To determine semen kinematic characteristics, we measured motility (MOT), straight-line velocity (VSL), curvilinear velocity (VCL), average path velocity (VAP), linearity (LIN), straightness (STR), amplitude of lateral head displacement (ALH), and beat cross frequency (BCF) of spermatozoa in boars. In order to detect single nucleotide polymorphisms (SNPs), we extracted genomic DNA from multiple Duroc boars, and then subsequently used them in sequencing reactions. As a result, three SNPs were detected in the intronic region of ZAR1 gene (g.2435T>C in intron 2, g.2605G>A and g.4633A>C in intron 3 ). SNPs g.2435T>C and g.2605G>A were significantly associated with MOT (p<0.01) and VSL (p<0.05), and g.4633A

• Korean Journal of Pharmacognosy
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• v.12 no.3
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• pp.119-124
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• 1981

In our country, in order to cure diseases, a large number of crude drug preparations has been available. Nevertheless, the development of crude drug preparations have been inhibited, because the quality control is not completed so far. Therefore, we have eontinued on studing the quality control method by Zig-zag TLC. profile analysis. The water extract of 'Samyo-Tang' and componental crude drug (Glycyrrhizae Radix, Ephedrae Herba, Armenicae Semen) were developed on Silica gel $60F_{254}\;plate\;(E.\;Merck)$ useing elution solvent. The developed plate were examined useing Dual Wavelength Zig-zag Scanner (Shimadzu). According to the results of the experiment, it could be summarized as follow: 1) Original patterns of TLC profiles of 'Samyo-Tang' componental crude drug and mixing two crude drugs of 'Samyo-Tang' were observed. 2) Original patterns TLC profile of each extract after spraying with 2% ninhydrine were observed. 3) In the extract of addition and subtraction of Ephedrae Herba, peak area of Rf 0.48 and Rf 0.60 were varied quantitatively.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.19 no.9
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• pp.116-123
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• 2018

The sperm quality is determined by the kinetic characteristics and acrosome integrity of the sperm. In the previous studies, analysis of semen quality had large errors because those experiments by using microscope had been conducted by people. In recent years, the molecular biological methods have been newly developed to complement the previous techniques. The ZAR1 gene is known to be a gene that affects early embryonic development in vertebrates, but there is no study of the association with semen. In this study, we analyzed the association between the kinetic characteristics and ZAR1 single nucleotide polymorphism (SNP) genotype. To detect the SNPs, we performed sequencing using genomic DNA from the whole bloods of Duroc pigs. We identified an SNP in the ZAR1 gene g.2540T>C. ZAR1 SNP genotypeing in 105 pigs revealed that the major and minor alleles were T and C, respectively. After we analyzed the association between the kinetic characteristics of sperm and the ZAR1 SNP genotype, we found a significant association in MOT (p<0.01), VSL (p<0.05) of the kinetic characteristics in the Duroc boars. It was confirmed that the boars with T allele were lower in MOT and VSL than C allele. Therefore, pigs with C allele are judged to be better at the MOT and VSL of semen. Based on these results, ZAR1 SNP genotyping may be a useful molecular biomarker to improve semen quality by applying molecular breeding technology.

• Journal of Embryo Transfer
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• v.26 no.4
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• pp.257-263
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• 2011

BPA, a diphenyl compound containing groups, that make it structurally similar to synthetic estrogen and is considered as one of the major endocrine disruptors. Silymarin has extensively been used to prevent and/or alleviate some human disease, especially for the treatment of adverse liver conditions. It has an antioxidative efficacy and cancer preventive efficacy. Therefore, we examined the hypothesis that silymarin can inhibit BPA-induced toxicity in boar sperm duing in vitro storage. Sperm characteristics (motility, viability, membrane integrity and mitochondrion activity) in semen exposed to BPA (10~200 uM) were sharply lowered, while it increase in a dose and time dependent manner due to silymarin addition (50~200 uM) into semen extender in the presence of BPA (100 uM). All of the evaluated characteristics were gradually improved in the groups that were treated with silymarin (50~200 uM) in the presence of BPA (100 uM) in comparison to BPA 100 uM alone group, irrespective of incubation periods (3 and 6 h). These results demonstrate that silymarin can ameliorate the toxicity of BPA on boar sperm characteristics during in vitro storage, suggesting that silymarin indirectly act as an antioxidant.

• Clinical and Experimental Reproductive Medicine
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• v.43 no.2
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• pp.97-101
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• 2016

Objective: Growth hormone and its mediator, insulin-like growth factor-1 (IGF-1), have been suggested to exert gonadotropic actions in both humans and animals. The present study was conducted to assess the relationship between serum IGF-1 concentration, seminal plasma concentration, and sperm parameter abnormalities. Methods: A total of 79 men were enrolled in this study from December 2011 to July 2012 and were prospectively analyzed. Patient parameters analyzed included age, body mass index, smoking status, urological history, and fertility history. Patients were divided into four groups based on their semen parameters: normal (A, n=31), abnormal sperm motility (B, n=12), abnormal sperm morphology (C, n=20), and two or more abnormal parameters (D, n=16). Patient seminal plasma and serum IGF-1 concentrations were determined. Results: Patient baseline characteristics were not significantly different between any of the groups. The serum IGF-1 levels in groups B, C, and D were significantly lower than the levels in group A; however, the seminal plasma IGF-1 levels were not significantly different between any of the groups. Conclusion: Men with abnormal sperm parameters had significantly lower levels of serum IGF-1 compared with men with normal sperm parameters. Seminal plasma IGF-1 levels, however, did not differ significantly between the groups investigated here. Further investigations will be required to determine the exact mechanisms by which growth hormone and IGF-1 affect sperm quality.

• Journal of the Korean Applied Science and Technology
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• v.33 no.4
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• pp.693-697
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• 2016

In this study, the electron donating ability(EDA) and total polyphenol content of herbal wine were examined. The herbal wine was obtained from extract concentration to evaluate its functional properties. The herbal wine were screened for their potential antioxidant activities using test such as electron donating ability(EDA) and total polyphenol content. The electron donating ability(EDA) were $21.81{\pm}0.56$ in herbal wine 15% and $40.45{\pm}1.60$ in herbal wine 35%. As the extract concentration was increased the electron donating ability(EDA) were significantly increased(p<0.05). The total polyphenol contents were measures $113.89{\pm}1.79{\mu}g\;GAE/m{\ell}$ in herbal wine 15%, $274.24{\pm}0.71{\mu}g\;GAE/m{\ell}$ in herbal wine 35%. As the extract concentration was increased the total polyphenol contents were significantly increased(p<0.05). Also, the total polyphenol contents were measures $61.75{\mu}g\;GAE/m{\ell}$ in herbal wine, the higher.

• Journal of Environmental Health Sciences
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• v.48 no.4
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• pp.244-253
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• 2022

Background: The indiscriminate use of pesticides in Nigeria may have harmful effects on reproductive health of farmers. Objectives: This study assessed the awareness of reproductive health, serum follicle stimulating hormone (FSH), luteinizing hormone (LH), testosterone, estradiol, progesterone and sperm characteristics of male farmers occupationally exposed to pesticides. Methods: Eighty four male farmers were recruited for the study. Structured questionnaire was used to obtain the socio-demographic data. Blood and semen samples were collected from the subjects in the morning for hormonal assays and semen analysis using enzyme linked immunosorbent assay (ELISA) method and SQAV sperm quality analyzer. Data were analyzed using chi square, Student's-t-test, and Regression analysis. Results: Serum FSH (p<0.01), LH (p<0.005) and Estradiol (p<0.001) were significantly higher while prolactin (p<0.02) and testosterone (p<0.001) were significantly lower among pesticides exposed farmers than nonexposed subjects. Some 34/84 (40.5%) of the pesticides exposed farmers had serum testosterone levels below the lower limit of the reference range. Those with low testosterone levels (p<0.001), also had FSH (p<0.05), LH (p<0.001) and Estradiol (p<0.002) significantly lower than those with normal testosterone levels. The sperm count among pesticides exposed farmers; total motility and percentage morphology were significantly lower than non-pesticides exposed subjects. Some 14/84 (16.7%) of the pesticides exposed farmers had sperm count below 15 million/mL (oligozoospermia). More than 70% of the farmers were not aware of the reproductive health risks associated with pesticides and only 23.8% of the farmers were using protective devices. Conclusions: Deliberate efforts to improve awareness, knowledge, personal hygiene, and interventions necessary to lessen both pesticides exposure and health risks by adopting safe practices are suggested.