• 한국동물생명공학회지
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• 제37권4호
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• pp.255-265
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• 2022

The cellular communication network factor (CCN) family proteins regulate many biological events such as angiogenesis, tumor growth, placentation, implantation, and embryogenesis. The expression and function of CCN1, CCN2, and CCN3 at the maternal-conceptus interface are established in humans and rodents, but little is known about the role of CCN4 to CCN6 in the reproductive organs in any other species. Several studies in transcriptome analysis in pigs have shown that the expression of CCN4 and CCN6 increases in the endometrium during early pregnancy. However, their expression, regulation, and function in the endometrium throughout the estrous cycle and pregnancy have not been fully understood in pigs. Thus, we determined the expression, localization, and regulation of CCN4 and CCN6 during the estrous cycle and at the maternal-conceptus interface in pigs. We found that the levels of CCN4, but not CCN6, changed during the estrous cycle. The levels of CCN4 were greater during mid- to late pregnancy than in the early stage, and the levels of CCN6 were greatest on Day 15 of pregnancy. CCN4 and CCN6 were detected in conceptus tissues during early pregnancy and in chorioallantoic tissues during the later stage of pregnancy. CCN4 mRNA was mainly localized to epithelial cells, CCN6 mRNAs to epithelial and stromal cells in the endometrium. In endometrial explant cultures, CCN4 expression was increased by progesterone, and CCN6 expression by interferon-𝛾. These results suggest that CCN4 and CCN6 may play roles in the establishment and maintenance of pregnancy by regulating the endometrial epithelial cell functions in pigs.

• Clinical and Experimental Reproductive Medicine
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• 제49권4호
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• pp.248-258
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• 2022

Objective: This research investigated the effects of human chorionic gonadotropin (HCG)-producing peripheral blood mononuclear cells (PBMCs) on the implantation rate and embryo attachment in mice. Methods: In this experimental study, a DNA fragment of the HCG gene was cloned into an expression vector, which was transfected into PBMCs. The concentration of the produced HCG was measured using enzyme-linked immunosorbent assay. Embryo attachment was investigated on the co-cultured endometrial cells and PBMCs in vitro. As an in vivo experiment, intrauterine administration of PBMCs was done in plaque-positive female mice. Studied mice were distributed into five groups: control, embryo implantation dysfunction (EID), EID with produced HCG, EID with PBMCs, and EID with HCG-producing PBMCs. Uterine horns were excised to characterize the number of implantation sites and pregnancy rate on day 7.5 post-coitum. During an implantation window, the mRNA expression of genes was evaluated using real-time polymerase chain reaction. Results: DNA fragments were cloned between the BamHI and EcoRI sites in the vector. About 465 pg/mL of HCG was produced in the transfected PBMCs. The attachment rate, pregnancy rate, and the number of implantation sites were substantially higher in the HCG-producing PBMCs group than in the other groups. Significantly elevated expression of the target genes was observed in the EID with HCG-producing PBMCs group. Conclusion: Alterations in gene expression following the intrauterine injection of HCG-producing PBMCs, could be considered a possible cause of increased embryo attachment rate, pregnancy rate, and the number of implantation sites.

• 한국임상수의학회지
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• 제28권6호
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• pp.610-612
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• 2011

9 년령 암컷 콜리견이 복부 팽만, 식욕 부진 및 변비를 주증으로 본원에 내원하였다. X-ray, 초음파 및 CT 검사를 통하여 복강내 연부조직종괴를 확인하였으나 세포학적 검사에서 종양 세포는 관찰 할 수 없었다. 탐색적 개복술을 계획하였다. 크고 둥근 모양의 종괴가 우측 자궁각에 위치하는 것을 확인하고 난소자궁적출술을 실시하였다. 종괴의 크기는 $40{\times}25{\times}18$ cm 였으며 무게는 9.5 kg 이었다. 조직학적 검사를 통해 낭상자궁내막증식증(CEH)으로 진단하였다. CEH는 자궁 내용물이 감염되거나 자궁 축농증으로 이어짐에도 불구하고 임상 증상이 없는 경우가 많으므로 정확한 진단이 어렵다. 본 증례는 자궁 축농증을 동반하지 않은 매우 큰 크기의 국소성 낭상자궁내막증식증의 드문 예이다.

• Asian-Australasian Journal of Animal Sciences
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• 제25권1호
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• pp.44-51
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• 2012

Calcium ions play an important role in the establishment and maintenance of pregnancy, but molecular and cellular regulatory mechanisms of calcium ion action in the uterine endometrium are not fully understood in pigs. Previously, we have shown that calcium regulatory molecules, transient receptor potential vanilloid type 5 (TRPV6) and calbindin-D9k (S100G), are expressed in the uterine endometrium during the estrous cycle and pregnancy in a pregnancy status- and stage-specific manner, and that estrogen of conceptus origin increases endometrial TRPV6 expression. However, regulation of S100G expression in the uterine endometrium and conceptus expression of S100G has been not determined during early pregnancy. Thus, we investigated regulation of S100G expression by estrogen and interleukin-$1{\beta}$ (IL1B) in the uterine endometrium and conceptus expression of S100G during early pregnancy in pigs. We obtained uterine endometrial tissues from day (D) 12 of the estrous cycle and treated with combinations of steroid hormones, estradiol-$17{\beta}$ ($E_2$) and progesterone ($P_4$), and increasing doses of IL1B. Real-time RT-PCR analysis showed that $E_2$ and IL1B increased S100G mRNA levels in the uterine endometrium, and conceptuses expressed S100G mRNA during early pregnancy, as determined by RT-PCR analysis. To determine if endometrial expression of S100G mRNA during the implantation period was affected by the somatic cell nuclear transfer (SCNT) procedure, we compared S100G mRNA levels in the uterine endometrium from gilts with SCNT-derived conceptuses with those from gilts with conceptuses derived from natural mating on D12 of pregnancy. Real-time RT-PCR analysis showed that levels of S100G mRNA in the uterine endometrium from gilts carrying SCNT-derived conceptuses was significantly lower than those from gilts carrying conceptuses derived from natural mating. These results showed that S100G expression in the uterine endometrium was regulated by estrogen and IL1B of conceptus origin, and affected by the SCNT procedure during early pregnancy. These suggest that conceptus signals regulate S100G, an intracellular calcium transport protein, for the establishment of pregnancy in pigs.

• 한국수정란이식학회지
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• 제27권4호
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• pp.245-252
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• 2012

The purpose of the present study was to investigate the effect of IFN-${\tau}$ on prostaglandin synthesis, cyclooxygenase-2 (COX-2) gene expression in vitro and concentration of progesterone (P4) in endometrial cells. Epithelial and stromal cells cultured in vitro were isolated from bovine endometrium and stimulated with increasing doses of IFN-${\tau}$ (0, 0.02, 0.2 and 2 ug/ml). Human chorionic gonadotropin (hCG, 1.5 IU/ml) was used as a positive control. Prostaglandin $E_2$ and $F_{2{\alpha}}$ levels in the culture media were analyzed by enzyme immunoassays and total RNA was extracted from the cells for RT-PCR. P4 concentrations of blood samples were assayed by chemiluminescent immuno assays system. In epithelial cells, COX-2 gene expression was increased in the presence of IFN-${\tau}$ (p<0.05), but it was not significantly different in all groups of stromal cells except for 2 ug/ml IFN-${\tau}$ group (p<0.05). Although IFN-${\tau}$ did not affect $PGE_2$ and $PGF_{2{\alpha}}$ production in epithelial cells, it decreased $PGE_2$ and $PGF_{2{\alpha}}$ production significantly in stromal cells (p<0.05). In vivo experiment, blood concentration of P4 was significantly increased after addition of IFN-${\tau}$ (1 ug/ml). The results indicate that PG production was mediated by COX-2 expression in stromal cells but it was not affected in epithelial cells and this suggest that treatment of IFN-${\tau}$ could improve the implantation environment of uterine by maintenance of high P4 concentration.

• Radiation Oncology Journal
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• 제26권3호
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• pp.149-159
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• 2008

목 적: 본 연구는 자궁내막암 병기 $1{\sim}2$ 환자에 수술 후 방사선치료의 결과와 예후인자를 분석하여 향후 치료원칙을 결정하는 데 도움을 얻고자 시행하였다. 대상 및 방법: 1991년 1월부터 2005년 12월까지 이화여자대학교 의과대학부속병원에서 자궁내막암으로 진단받고 수술 후 방사선치료를 받은 병기 $1{\sim}2$환자를 대상으로 후향적 분석을 하였다. 전체 환자는 35명이었으며 17명은 자궁절제술과 양측 난소제거술을 시행하였고 18명은 완전한 수술적 병기의 수술을 시행하였다. 골반강 방사선치료를 받은 환자는 17명, 질강내근접치료를 받은 환자는 12명, 골반강 방사선치료와 질강내근접치료를 받은 환자는 6명이었다. 결 과: 추적기간의 중앙값은 54개월이었다. 전체 환자의 5년 생존율은 91.4%, 무병생존율은 81.7%이었다. 저위험군, 중간위험군, 고위험군의 위험군에 따른 5년 생존율은 각각 100%, 100%, 55.6%이었으며 무병생존율은 각각 100%, 70.0%, 45.7%이었다. 국소재발된 환자는 없었으며 5명(14%)에서 원격전이가 발생하였으며 호발부위는 폐, 뼈, 간, 부신, 전복강의 순이었다. 원격전이에 대한 단변량 통계분석에 따르면 위험군, 조직학적 세포형태와 조직학적 등급이 유의성이 있었으며, 다변량 통계분석에 따르면 조직학적 세포형태로서 유두형, 장액형, 그리고 투명세포암은 자궁내막양암과 샘암편평암에 비하여 원격전이율이 높고 생존율이 낮았다. 골반강 방사선치료에 의한 후유증은 장염이 30%에서 나타났으며 다음으로 직장염이었으나 대부분 등급 $1{\sim}2$로 등급 $3{\sim}4$의 후유증은 발생하지 않았다. 결 론: 수술 후 골반강 방사선치료 혹은 질강내근접치료를 시행한 자궁내막암 병기 $1{\sim}2$환자들의 경우 저위험군과 중간위험군은 국소제어율과 생존율이 높았으나, 고위험군은 골반강 방사선치료를 시행하여 국소제어율은 높으나 원격전이율이 높고 생존율이 낮았다. 따라서 고위험군은 방사선치료와 동시에 항암화학요법이 요구된다. 중간 위험군의 더 효율적인 치료를 위하여 많은 환자를 대상으로 골반강 방사선치료와 강내근접치료를 비교하는 전향적 무작위연구가 필요하다.

• 한국콘텐츠학회:학술대회논문집
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• 한국콘텐츠학회 2013년도 춘계 종합학술대회 논문집
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• pp.361-362
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• 2013

As one of the steps toward understanding how the plant is well adapted to strongly saline habitats, the purple pigment compound that is accumulated in Suaeda japonica was extracted and characterized. The extracted pigment compound exhibited typical characteristics of betacyanin that were represented by water solubility, pH- and temperature-dependent color changes, sensitivity to light, UV-Vis spectra, and gel electrophoretic migration pattern. LC-MS of the extracted pigment compound showed the presence of two major protonated molecular ions ($[M+H]^+$) at m/z 651.1 and m/z 827.1. According to the DPPH assay, it was found to have an antioxidant activity that is linearly increased in proportion to the reaction time for up to 30 min, and the activity was comparable to that of control BHA at 9.0 mg/ml. The cytotoxic activity against several tumor cell lines was also examined following the MTT assay. The significant growth inhibitory effect was observed on two tumor cell lines, SW-156 (human kidney carcinoma) and HEC-1B (human endometrial adenocarcinoma). Probably, the pigment compound may function as an osmolyte to uphold halotolerant physiological processes in saline environment.

• 한국자원식물학회지
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• 제31권4호
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• pp.342-354
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• 2018

Suaeda japonica is a halophytic plant that lives in mudflat at intertidal zone of western and southern coastal areas of Korea. The seawater-living plants showed a purple color during their whole life. In contrast, freshwater-living plants displayed a green color in leaves. When seawater-living plants were transferred to potting soil, the purple color was gradually changed to green in the leaves. The extracted purple pigment compound exhibited typical characteristics of betacyanin that were represented by water solubility, pH- and temperature-dependent color changes, sensitivity to light, UV-Vis spectra, and gel electrophoretic migration pattern. The LC-MS analysis of the extracted pigment compound showed the presence of two major protonated molecular ions ($[M+H]^+$) at m/z 651.1 and m/z 827.1. Antioxidant activity of the pigment compound was determined using stable free radical DPPH assay. It was found to have an antioxidant activity that is linearly increased in proportion to the reaction time for up to 30 min, and the activity was comparable to that of control BHA at 9.0 mg/ml. The anticancer activity against several tumor cell lines was also examined following the MTT assay. The significant growth inhibitory effect was observed on two tumor cell lines, SW-156 (human kidney carcinoma) and HEC-1B (human endometrial adenocarcinoma). Probably, the pigment compound may function as an osmolyte to uphold halotolerant physiological processes in saline environment.

• Asian-Australasian Journal of Animal Sciences
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• 제24권7호
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• pp.919-928
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• 2011

The pig exhibits true epitheliochorial placentation, where the fetal membrane maintains attachment throughout pregnancy but does not invade into the maternal uterine endometrium. Accordingly, the expression and function of cell adhesion molecules are very important for embryo implantation and the establishment of pregnancy. In our recent microarray analysis, we found that activated leukocyte cell adhesion molecule (ALCAM) was expressed in the uterine endometrium during pregnancy in pigs. To better understand the roles of ALCAM in the establishment and maintenance of pregnancy, we examined ALCAM expression in the uterine endometrium during the estrous cycle and pregnancy in pigs. Real-time RT-PCR analysis showed that ALCAM was differentially expressed in the uterine endometrium during the estrous cycle and pregnancy, with the highest levels on D12 of pregnancy. ALCAM mRNA was localized to the luminal and glandular epithelial cells and to the trophectoderm of conceptuses during early pregnancy. The steroid hormones estrogen and progesterone had no effect on ALCAM expression in an endometrial explant culture study. Further, we found that ALCAM expression in the uterine endometrium from gilts with somatic cell nuclear transfer-derived embryos was not different from that in gilts with embryos from natural mating. ALCAM was expressed in a pregnancy stage- and cell type-specific manner in the uterine endometrium and conceptuses during pregnancy. These findings suggest that ALCAM may play a role in the establishment of pregnancy. Further analysis of ALCAM will provide insight into the implantation process and establishment of pregnancy in pigs.

• 대한한방부인과학회지
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• 제20권3호
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• pp.1-12
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• 2007

Purpose: Uterine leiomyomas (fibroids) are common estrogen-dependent uterine tumors. Houttuynia cordata thunberg has cancer-preventing properties and often used in Chinese medicine. In the present study we used Houttuynia cordata thunberg to determine its effect on cell proliferation and apoptosis in human uterine leiomyoma cells. Methods: Primary cultured human uterine leiomyoma cells were treated with Houttuynia cordata thunberg. Cell viability analysis was analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTS) assay and Flow cytometry was performed to ascertain the effects Houttuynia cordata thunberg. Expression of cell cycle related proteins and apoptosis related proteins were evaluated by Western blot analysis. Results: Cell viability was significantly influenced by Houttuynia cordata thunberg treatment in a dose-dependent manner in leiomyoma cells compare to normal myometrial cells. Flow cytometric analysis showed that Houttuynia cordata thunberg induced Sub G1 arrest. DNA fragmentation assay was carried out and apoptosis was detected. Activation of caspase-3, down-regulation of Bcl-2, with concomitant increase in p21 was observed. Houttuynia cordata thunberg treatment of uterine leiomyoma cellsresulted in a concentration-dependent cell death induced via the caspase dependent mechanism. Conclusion: These results suggest that Houttuynia cordata thunberg treatment in uterine leiomyoma cells leads to growth inhibition and induced apoptosis. These results suggest that Houttuynia cordata thunberg will be a promising agent for use in therapeutics agents against human uterine endometrial cancer.