Background: Water deer and sika deer, which breed in the wild environment, are known to have similar reproductive physiology mechanisms. Therefore, this study aimed to analyze the differences in uterine development between water deer and sika deer during estrus. Methods: MMPs and uterine development-related factors were analyzed and morphological differences were compared in the uterus of sika deer captured near Russia near Korea and water deer captured in the wild in Korea. Results: In terms of morphological differences in the uterus, the glands that form villus within the endometrium of the water deer were newly developed, and the formation of small glands was high, but the villus and glands of the sika deer were expanded, and the stroma zone in the myometrium was higher than that of the water deer. Development has increased. Additionally, the expression of PAPP-A and VEGF factors was increased in the endometrium of water deer than in sika deer, but the actions of MMPs were increased in sika deer. Conclusions: As a result of this study, there is a significant difference in the development of glands in the endometrium of water deer and sika deer during estrus, and it is believed that there is a significant difference in the development of the uterus due to the physiological effects of estrus between water deer and sika deer. Additionally, it is believed that there will be differences in the timing at which pregnancy can be decided.

Background: A breast cancer is the second leading cause of cancer death in women worldwide and among different types of breast cancers, triple-negative breast cancer (TNBC) has a poor prognosis. Methods: We investigated the potential of ginsenoside compound K (CK), an active ingredient in the bio-transformed ginsenoside, to be used as a therapeutic ingredient by examining the effects of CK on cell proliferation, apoptosis, and cancer-related gene expressions in breast cancer cells. Results: From the results of treating MCF-7, an ER and PR-positive breast cancer cells, and MDA-MB-231 (TNBC) with CK at a concentration of 0-100 μM, the half maximal inhibitory concentration (IC50) values for each cell were 52.17 μM and 29.88 μM, respectively. And also, it was confirmed that cell migration was inhibited above the IC50 concentration. In addition, fluorescence analysis of Apoptosis/Necrosis showed that CK induced apoptosis rather than necrosis of breast cancer cells. Through qPCR, it was confirmed that the expression of genes related to apoptosis and cell cycle arrest was increased in CK-treated breast cancer cells, and it acted more effectively on TNBC. However, the expression of genes related to tumor invasion and metastasis is also increased, so it is necessary to consider the timing of application of CK as a potential therapeutic anticancer compound. Conclusions: CK showed a stronger inhibitory effect in TNBC with poor prognosis but considering the high tumor invasion and metastasis-related gene expression, the timing of application of CK should be considered.

Background: This study has mainly focused on finding pharmacological effects of ginsenosides that can reduce the unwanted side effects of the cytotoxic anticancer drugs and are highly effective on prostate cancer, colorectal cancer, liver cancer, hormone-dependent breast cancer, triple-negative breast cancer, and brain cancer (neuroblastoma). Methods: Minor and rare ginsenosides (GS) of Rh2 which have a high absorption ability and excellent pharmacological actions were treated with the 6 different types of cancer cell lines and their anticancer activities were investigated by analyzing gene expressions associated with various cancers through qPCR and other relevant methods. Results: In cancer cells exposed to Rh2, cell viability and cell migration were reduced, and apoptosis was induced. Each cancer cell was divided into three groups according to the cell proliferation response by Rh2; 1) A group in which the cell viability decreases inversely to an increase in Rh2 treatment concentration; 2) A group in which the cell viability rapidly decreases in Rh2 treatment above a certain level of concentration; 3) A group in which the cell viability was not suppressed below 20-30% even with 100 μL of Rh2, the highest concentration used in this study. Conclusions: It was shown that Rh2 has a significant effect on inhibiting the proliferation of prostate cancer cells and hormone-dependent breast cancer cells.

Background: Research on the reproductive physiology of Water and Sika deer, an endemic in Korea, still needs to be completed. This study analyzed the ovarian development and morphological characteristics of wild Water deer and Sika deer. Methods: Water deer and Sika deer ovaries were collected from the Korean Peninsula and Russia–Korean Peninsula border during the estrus and pregnancy seasons, respectively. And, morphological and physiological analysis and immunohistochemistry were conducted to confirm the detection of Ca2+ and assess the morphological changes in the ovaries. Results: The results of morphological analysis of ovaries during pregnancy and estrus, the development of the corpus luteum and follicles of Water deer showed similar patterns to other mammals. In contrast, the corpus luteum of Sika deer differed in tissue morphology and composition from Water deer. Ca2+ related to tissue metabolism was detected in the theca cells zone of Water deer on the estrus and was highly detected in the luteum cells zone during pregnancy. The hormone receptor protein expression patterns were generally higher in the ovaries of Water deer on the estrus and the pregnancy than in Sika deer. The expression of LH receptor was relatively low in the lutein cell zone, unlikely that of Water deer. The expression of VEGF was also different from Water deer, and the response in Sika deer was relatively very low compared to Water deer in expressing all proteins-related development. Conclusions: Therefore, the results of the study were shown that the composition of the corpus luteum of Sika deer is not clear compared to Water deer, and there are many differences in the functional and morphological formation of the corpus luteum.

The sheep can be reproduced by natural mating as well as applied reproductive biotechnology, embryo transfer (ET). However, this method in sheep is influenced by several factors such as season, photoperiod, latitude, temperature, nutrition, and breed. In addition, there is still less research on assisted reproductive technologies in small ruminants, compared to other livestock species such as cattle and pigs. Because there has been a need for an optimization and a continuous improvement of ET techniques in small ruminants. the main objective of this study was to evaluate the conception rate obtained after ET in Mongolian sheep (Dorper breed). After embryo recover, code 1 and 2 embryos (morula or blastocyst stage) for ET in the present study were 63% (63/100) and 24% (24/100), respectively. Then Each single embryo was transferred to a synchronized recipient who prepared by estrous synchronization protocol with fluorogestone acetate-cloprostenol sodium. The results demonstrated that an average conception rate and lambing rate was 35.6% (31/87) and 33.3% (29/87), respectively. Further study is still necessary, but these results indicated that single embryo of Mongolian sheep with the present protocol was enough to conducting ET when the genetically superior sheep were necessary to be expanded.

This study aimed to determine whether hormonal hypersecretion could cause morphological problems in the mouse vagina and affect the ovaries and nearby extra uterine organs. All mice were synchronized to estrus before the experiment. Then human chorionic gonadotropin (hCG), progesterone, and testosterone were continuously administered for about 6 days to maintain hormone hypersecretion, and then morphological changes were analyzed, and Matrix metalloproteinases (MMPs) activity and Casp-3 expression were evaluated. As a result of the analysis, in the case of hCG, the morphological change did not show a significant difference from the vagina of normal estrus. In the case of progesterone, changes were observed in the mucosa zone and basal membrane, and it was confirmed that the activity of MMPs was increased in squamous epithelium cells. On the other hand, in the case of testosterone, overall changes in vaginal tissues were observed, and MMPs activity was increased to a very high level in all sections. The expression of Casp-3 was also the highest compared to other groups. Therefore, as a result of this study, it is thought that hormone hypersecretion affects the morphological changes of the vagina other than the ovaries and uterus and induces the activity of MMPs to cause morphological degeneration of tissues.

In this study, to analyze whether artificial regulation of apoptosis in the development of somatic cells can affect the stable growth and development of cells, 20 alpha-hydroxysteroid dehydrogenase (20α-HSD) and rapamycin were treated to induce apoptosis and autophagy in the both skin and muscle cells. Respectively, and 3-methyladenine was supplemented to inhibit cell death. Our results show that stimulation with rapamycin activated autophagy in both types of cells, but increased apoptosis more than autophagy in the case of skin cells. These results indicate that there was a difference in the expression of survival factors and cell development depending on the type of cell. In particular, in the expression of autophagy-related gene (MAP1LC3A) was higher than that of Casp-3, an apoptosis factor. Furthermore, cell development was the highest in all cell groups cultured by artificially inducing autophagy, however the lowest in the apoptosis-inhibited group. Especially, the noteworthy result of this study was that when apoptosis was induced using 20α-HSD, it was possible to induce apoptosis in both skin and muscle cells. Therefore, the main point of this study is that apoptosis induced during cell culture plays a pivotal role in cell remodeling.

In the early development of parthenogenetic embryo, cytoplasm and nucleic acid fragmentation may be a cause of lower embryo development. The purpose of this study was to evaluate whether embryonic development and apoptosis factors can be reduced by controlling the in-vitro culture environment by the addition of hormones, pregnancy serum and uterine milk. Our study showed that the activity of Casp-3 increased within the cytoplasm when artificially used hormones to induce the incubation environment, and PCNA's manifestation was low. However, the addition of pregnant serum appeared to lower the Casp-3 activity compared to the other groups. In addition, MMP-9 activity was increased and early embryo development and cytoplasmic fidelity were also increased. Therefore, the results of the present study showed that the use of gestational serum in the development of parthenogenetic embryo inhibit apoptosis and increases cytoplasmic reorganization by natural environmental control in in vitro culture.

In this study, we investigated whether infusion of colorectal cancer cell line and PMSG could increase endometrial cancer. As a result, our study confirmed that the injection of colorectal cancer can cause inflammation and cancer in the uterus and increase the VEGF gene in the uterus. The study also found that endometrial cancer was associated with PMSG.

Our study has analyzed whether inappropriate gonadotropin secretion affects the morphological changes due to the activation of intrauterine MMP. Methods A total of each 6 mice were injected with PMSG, Progesterone, and Androgen in 5 IU of intraperitoneal injection every 2 days after estrus synchronization, and morphological and MMPs expression patterns were compared after inducing hormone secretion. Also, cell survival and death related genes were compared and analyzed. The endometrium was highly developed in the PMSG, and the androgen was not developed at all. In particular, the diameter of the uterus of the Androgen group was also very narrow. MMPs activity assay in the case of PMSG was confirmed that showed low activity, whereas, progesterone and androgen In showed high activity and, in particular, very high activity of MMPs in the case of androgen in glandular cell. The expression of VEGF in the tissues of each group was different from that of MMPs. In the PMSG group, the activity of VEGF was increased in both the Myo-metrium and the endo-metrium, whereas the progesterone group showed low overall expression in the endo-metrium. Therefore, the present study showed that the activities of the endo-metrial cells and the restructuring of the endometrial cells differed according to the type of the abnormal secretory hormone. In particular, the secretion of androgen increased the activity of MMPs throughout the uterus, The endo-metrial epithelial cells are affected by the progesterone group. In conclusion, this study suggests that inappropriate gonadotropin secretion increases the functional changes of the uterus and this reconstruction may be caused by increased activity of MMPs

The expression of MMPs in the development of the fertilized egg has a very important role in cell configuration. Objective To evaluate the clinical, the effect of differentially expressed MMPs on serum and serum - free medium on the maturation of blastocysts. The expression patterns of MMPs in serum and serum-free medium were compared at 6 h, 18 h and at the blastocyst stage using real-time PCR, ELISA and immunofluorescence. The results showed that the expression of MMPs was increased in the embryos of the serum medium, as a result of analysis of MMPs and TIMPs, MMP-2 was expressed in the cytoplasm of embryos in the serum-free medium, And it was found to be higher in expression than MMP-9. The serum medium was different from the bloodless badge: overall, TIMPs showed a higher expression in the ovarian cells than cyanosis, and TIMP-3 was more pronounced. Development rate of blastocyst according to in vitro culture method was higher than that of serum - free medium (61.22% 60/98) and serum - free medium (48.28% 28/58). Analysis of the protein release locations of MMPs and TIMPs showed that MMPs and TIMPs are highly expressed in serum mediums, focusing on the inner cell mass. However, very low expression appeared in the tropoblast. On the other hand, serum - free medium showed different expression from serum medium and TIMPs expression was generally low. Therefore, in the case of serum media, the expression of MMPs is highly expressed in the cytoplasm of the fertilized egg, increasing the reconstruction of cells.

In the present study of this experiment was to understand the expression of apoptotic gene expression in the ovary of miniature pigs and pigs on the 15th day of estrus. Also the compare and analyze of programmed cell death type(Apoptosis and autophagy) expression pattern during mature oocyte on the miniature and normal pig cells. Analysis of mRNA gene expression of ovary in miniature and normal pigs on the 15th day of estrus showed that the expression of genes related to Autophagy (ATG13, MAP1LC3, Beclin1) was high in normal pigs but the expression of ATG1 and ATG5 genes was low. In addition, the expression of genes related to apoptosis (Casp-3, BAX) was high in the mini pigs, and the gene related to the LH hormone was high in the miniature pigs, whereas the expression of the gene related to the FSH hormone was high in the normal pigs. On the other hand, the result of muture oocyte on the miniature and normal pig cells is the expression of Casp-3 protein was moust high from treatment of FL+rapa (FSH+LH and Rapamycin) of the oocyte on the miniature pig cell. However, MAP1LC3 expression was higher in the oocytes of treatment of rapanycine treatment on the nomal pig cells. There was no gene expression in cumulus cells of matured oocytes in mini pig cells, whereas MAP1LC3 expression was higher in oocyte cumulus cells matured in normal pig cells. It was confirmed that the miniature and normal pigs showed different programmed cell death patterns, In the case of oocytes matured in miniature pig cells, MAP1LC3 gene expression was found to be low in spite of treatment with Autophagy regulator.

The purpose of this study is to expression pattern of melanogenesis associate genes on cultured melanocyte layer cells in Korean Brindle Cattle(Dark, Brindle and Yellow) were analyzed to evaluate the effects of sex hormones on the control of melanogenesis pathways. Korean Brindle Cattle(Dark, Brindle and Yellow) melanocyte in the skin cells was collected. after the addition of estrogen and testosterone, the culture was analyzed for expression of cell activity and melanin genes for 72 hours. For the analysis of estrogen in different coat color other than the melanogenesis-related genes it is increasingly yellow showed low expression. in particular, the cells of the brindle coat color is low active and expression of genes. However, the testosterone was low, the expression of cell activity inhibiting MMP-2. the expression of melanin genes actually showed a tendency to increase gradually, which is testosterone compared with the estrogen to be considered that affect the skin cell layer brindle coat color. In this study, stimulation with estrogen triggered the inhibition of MC1R of the melanocyte in brindle coat color, but testosterone is induced MC1R in melanocyte. Therefore, considered the eumelanin or phaeomelanin activation are controlled caused by differential expression of sex hormones on melanocyte in Korean Brindle Cattle.

ompared the expression of MMPs in these oocytes and cumulus cell throughout oocytes maturated. In an attempt to investigate the effect of MMP activation and inhibitors in total protein of cumulus cell and, oocytes during oocytes maturation, we examined and monitored the localization and expression of MMPs (MMP-2 and MMP-9), TIMPs (TIMP-2 and TIMP-3), as well as their expression profiles (Real-time PCR, Gelatin Zymography and ELISA). Our results that the bovine oocytes MMP-2 and MMP-9 level was significantly associated with the rate of maturity of oocytes (P<0.05). In cumulus cell, MMP-2 was highly expressed in all stages of the oocyte’s maturation. The final oocytes maturation exhibited strong gelatinase activity. There was no significant correlation between cumulus cell MMP-9 and the maturation rate of oocytes. However, for the oocyte cytoplasm MMP-9 expression was significant correlation to the maturation oocytes. There was no significant correlation between cumulonimbus cells MMP-9 and oocyte maturation rates; however, for oocyte cytoplasm, MMP-9 expression was significantly correlated with mature oocyte. However, the TIMP-1 and TIMP-2 protein expression patterns are not correlated with the maturation rate of the oocyte. Our results suggest that MMP different expression pattern may regulate the morphological remodeling of oocyte's in the cumulus cell. Further, the MMP-2 expression has a strong relation with a higher maturation rate of the oocyte.

The major focus of this study is to analyze the expression of bovine MMPs and to monitor their activity during the estrus cycle and pregnancy. During pregnancy, MMP-2 expression was detectable around 30 days but became insignificant by 60 days, then started to increase again around 90 days and reached the maximum at 250 days. The activity of MMP-2 protein changed in accordance with its expression level. As expected, the level of TIMP-2 exhibited a reverse pattern. About MMP-9, high level expression was observed as early as 30 days and gradually increase until 90 days. Then started to decrease after 250 days. Again, the sites of MMP-9 expression were similar to those of MMP-2. On the other hand, expression of TIMP-3 remained low until 90 days but showed a small and temporal increase around 250 days. In summary, expression of different MMPs were differentially regulated during estrus cycle and pregnancy. While the expression of MMP-2 was high in estrus cycle, MMP-9 slowly takes over with the progression of pregnancy. These results indicated that the luteal tissue perform distinct functions during pregnancy and estrus. Perhaps the activity of MMP-2 is required for the structural remodeling of luteum, resulting the suppression of P4 inflow from blood. On the other hand, steady maintenance of MMP-9 throughout luteal development is important for the activation of cell proliferation, maturation and angiogenesis.

The nature of molecular mechanisms governing embryo development is largely unknown, but recent reports have demonstrated that proper execution of programmed cell death is crucial for this process. The main objective of this study is to examine the mode of programmed cell death during nuclear transfer embryos development in porcine. In particular, the relative employment of two major pathways in programmed cell death; e.g. apoptosis (type I) and autophagy (type II) was compared. Oocytes use in the study was matured in vitro in the presence of 10% FBS maturation medium. After nuclear transfer embryos were cultured for each programmed cell death control factor [Cysteamine(Cyst : 0.4mM), 3-methyladenine(3MA : 2.5mM) and Rapamycin(RP : 100nM)] in TCM-199 medium supplemented with 0.1% BSA. In this study results of among the blastocysts development in 3MA; PCNA, MAP1LC3A and ATG5 RNA gene expression level increased in the order of IVF<Cyst < 3MA < RP. However Casp-3 and TNF-r RNA gene expression level decreased in the order of IVF < 3MA and RP< Cyst. The expression of mTOR showed a pattern opposite to that of MAP1LC3A. That is, its expression was the lowest in Cyst group. And next experiments analysis of MMP expression patterns. Analysed this MMPs enzyme activation to evaluate the effectiveness of high quality brastocyst culture in porcine. In this results of the enzymatic activity of MMP-2 and MMP-9 was assessed in culture, the level of active MMP-9 was higher expression in the medium of each 3MA and RP treatment group, with the level of another treatment group being relatively higher. These results suggest that the autophagy activation culture medium is more effective for stable and innovative nuclear transfer embryos development.

The coat color of mammals is determined by the melanogenesis pathway, which is responsible for maintaining the balance between black-brown eumelanin and yellow-reddish pheomelanin. The melanogenesis-associated genes controlling pigmentation act as a complex and interact with each other to cause phenotypic and genotypic variations in cattle. That the MC1R genotype of Korean native cattle with dark muzzle was e/e or E+/e, while the genotype of Korean native cattle with light muzzle was E+/E+, which is a variant of the MC1R genotype in the Korean native cattle. Especially, the MC1R expression type is shows how much pigmentation, important factor in deciding its status in the coat and nose colours. However, information regarding the coat or nose colours-associated gene regulation of korean cattle is not yet unknown. Therefore, in this study was to investigate the expression patterns of melanogenesis-associated genes in black dot nose(korea brindle cattle) and normal nose(korea native cattle). Using microarray clustering and real-time polymerase chain reaction techniques, we analysed that the expression of genes involved in the mitogen-activated protein kinase (MAPK) and Wnt signaling pathways is distinctively regulated in the dark and light muzzle tissues. Differential expression of tyrosinase was also noticed, although the difference was not as distinct as those of MAPK and Wnt. We hypothesize that emphasis on the MAPK pathway in the Korea brindle cattle induces eumelanin synthesis through the activation of cAMP response elementbinding protein and tyrosinase, while activation of Wnt signaling counteracts this process and raises the amount of pheomelanin in the native cattle. Regarding the increasing interest in the genetic diversity of cattle stocks, genes we identified for differential expression in the brindle cattle vs. native cattle may serve as novel markers for genetic diversity among cows based on the coat and muzzle color phenotype.

The tyrosinase (TYR) genes have been accepted as major genes involved in the plumage pigmentation of chickens. Tyrosinase (TYR) gene is located on chromosome 1 in chicken and it is composed of five exons and four introns. TYR gene is described as a key enzyme in melanin biosynthesis. Especially, most examples of color patterns in chicken have been due to differential in the tyrosinase gene. This study was conducted to the association of feather color and sequence polymorphism in the Tyrosinase(TYR) gene was investigated using Korean native chickens(red plumage, red-line plumage, Ogol = KNC) and white leghorn(WL). From WL and KNC breed analyses, 232 differential SNPs were detected in 4th exon and 4th intron of TYR gene respectively. The genotype frequencies for 50 SNPs were compared between KCR, KCRD and KCO represented homozygous SNP types in all the analyzed SNP positions while KNC displayed various SNP types. In this study, we conclude that the variation of a wild type sequence in intron 4 of the tyrosinase gene is pigmentation of the original native chickens in korean. This work was supported by a grant from the “Livestock Preservation of Genetic Resources", Rural Development Administration, Republic of Korea.

Recently, the consumption of chickens meat has been gradually increased in Korea. However, most of the chickens breeds in Korea were imported from overseas. This study was conducted to evaluate the genetic distances and single nucleotide polymorphism by the mtDNA D-loop control analysis method for the Korean native chickens(red plumage, red-line plumage, Ogol) and white leghorn. For the initial investigation of the relationships between Korean native chickens(red plumage(KCR), red-line plumage(KCRD), Ogol(KCO)) with white leghorn breeds, the sequences from D-loop control region in mitochondrial DNA (mtDNA) was used. The results from phylogenetic analysis indicated that both KNC and white leghorn breeds were classified well with wild duck breeds. However, KCR was not discriminated well with KCRD. The haplotype analysis indicated that KCR and KCO have ten different haplotypes with nineteen SNPs. Three haplotypes (haplotype 1, 2, 3, 5, 17) were shared both in KNC(KCR, KCRD) and KCO. On the other hand, haplotype 4, 6, 7 was appeared only KCR and haplotype 13, 16, 18 were identified only in KCRD population. In addition, haplotype 8, 9, 11, 12, 14, 15, 19 was appeared only KCO. With further verifications, the results presented here can be used for the on servation and commercialization of the Korean native chickens. This work was supported by a grant from the “Livestock Preservation of Genetic Resources", Rural Development Administration, Republic of Korea.

This study was conducted to evaluate the genetic distances and specific DNA makers by the randomly amplified polymorphic DNA (RAPD)-PCR method for the Korean native chickens(red plumage, red-line plumage, Ogol) and white leghorn. Genomic DNA was extracted from plumage from chickens after they were slaughtered. The extracted DNA was observed by nano-spectrometer. RAPD analysis was performed using 13 different primers. Statistical analysis was made for the estimation of the genetic distance among the chicken’s and the cluster tree was drawn by using MEGA 5.05 software. Genetic relations among them were determined by RAPD analysis. The polymorphic bands were observed 72% and the rest of 28% was monomorpic. The largest genetic distance (2.266) was found between the native chickens(red, red-line) and the ogol chickens by UPGMAP method and the closest distance was observed between the ogol in korean chickens as expected. The highest genetic distance between them was estimated 2.266 and in the dendrogram analysis, among I and II within cluster II, most of the ogol chickens were in IIB, indicating the expression of the ogol color could be due to original and the ancestral genetic crossing. Thus, this genetic distance can be useful as the differential genomic information in the normal(red plumage, red-line plumage) and ogol of korean native chickens. This work was supported by a grant from the “Livestock Preservation of Genetic Resources", Rural Development Administration, Republic of Korea.