• 한국동물생명공학회지
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• 제34권1호
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• pp.10-19
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• 2019

Morphology of cumulus-oocyte-complexes (COCs) at germinal vesicle (GV) stage as one of the evaluation criteria for oocyte maturation quality after in vitro maturation (IVM) plays important roles on the meiotic maturation, fertilization and early embryonic development in pigs. When cumulus cells of COCs are insufficient, which is induced the low oocyte maturation rate by the increasing of reactive oxygen species (ROS) in porcine oocyte during IVM. The ROS are known to generate including superoxide and hydrogen peroxide from electron transport system of mitochondria during oocyte maturation in pigs. To regulate the ROS production, the cumulus cells is secreted the various antioxidant enzymes during IVM of porcine oocyte. Our previous study showed that Mito-TEMPO, superoxide specific scavenger, improves the embryonic developmental competence and blastocyst formation rate by regulating of mitochondria functions in pigs. However, the effects of Mito-TEMPO as a superoxide scavenger to help the anti-oxidant functions from cumulus cells of COCs on meiotic maturation during porcine oocyte IVM has not been reported. Here, we categorized experimental groups into two groups (Grade 1: G1; high cumulus cells and Grade 2: G2; low cumulus cells) by using hemocytometer. The meiotic maturation rate from G2 was significantly (p < 0.05) decreased (G1: $79.9{\pm}3.8%$ vs G2: $57.5{\pm}4.6%$) compared to G1. To investigate the production of mitochondria derived superoxide, we used the mitochondrial superoxide dye, Mito-SOX. Red fluorescence of Mito-SOX detected superoxide was significantly (p < 0.05) increased in COCs of G2 compared with G1. And, we examined expression levels of genes associated with mitochondrial antioxidant such as SOD1, SOD2 and PRDX3 using a RT-PCR in porcine COCs at 44 h of IVM. The mRNA levels of three antioxidant enzymes expression in COCs from G2 were significantly (p < 0.05) lower than COCs of G1. In addition, we investigated the anti-oxidative effects of Mito-TEMPO on meiotic maturation of porcine oocyte from G1 and G2. Meiotic maturation and mRNA levels of antioxidant enzymes were significantly (p < 0.05) recovered in G2 by Mito-TEMPO ($0.1{\mu}M$, MT) treatment (G2: $68.4{\pm}3.2%$ vs G2 + MT: $73.9{\pm}1.4%$). Therefore, our results suggest that reduction of mitochondria derived superoxide by Mito-TEMPO may improves the meiotic maturation in IVM of porcine oocyte.

• 한국수정란이식학회지
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• 제25권3호
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• pp.171-177
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• 2010

Plasminogen activators (PAs) are serine proteases that convert plasminogen to plasmin. The PA/plasmin system has been associated with a number of physiological processes such as fibrinolysis, ovulation and fertilization. Although correlations have been reported between reactive oxygen species (ROS) and oocyte maturation, the relationship between PA activity and ROS is unknown. The present study was undertaken to determine the effects of cumulus cells on PA activity in matured porcine oocytes under xanthine (X)-xanthine oxidase (XO) system. When oocytes were matured under the X-XO system, the proportion of oocytes remaining GV stage was higher (p<0.05) in oocytes without cumulus cells. The incidence of degenerated oocytes was higher (p<0.05) in the X+XO ($11.1{\pm}6.1$ and $21.6{\pm}3.4%$) than in the control group ($2.9{\pm}1.8$ and $4.0{\pm}1.6%$). The proportion of TUNEL-positive oocytes and activity of caspase-3 were higher (p<0.05) in cumulus-free oocytes and oocytes exposed to ROS. Tissue-type plasminogen activator-plasminogen activator inhibitor (tPA-PAI) and tissue-type plasminogen activator (tPA) activity were detected in oocytes that were separated from cumulus-oocytes complexs (COCs) at 44 h of maturation culture, and only tPA was produced in oocytes that were denuded before the onset of maturation culture. On the other hand, the activities of PA were increased (p<0.05) when oocytes were cultured under the X-XO system. The higher activity of tPA was observed in denuded oocytes (DOs) underwent apoptotic changes by oxidative stress. In COCs, however, tPA-PAI as well as tPA activity was detected and apoptotic changes such as DNA cleavage or caspase-3 activation were not observed. These results suggest that tP A may be relevant to apoptotic cell death in porcine oocytes by oxidative stress.

• 동의신경정신과학회지
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• 제25권2호
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• pp.179-190
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• 2014

Objectives: This study was performed to review the research trends regarding the treatment of Postpartum Depression (P.P.D) in the Traditional Chinese Medicine (TCM). Methods: We searched articles in CNKI (China National Knowledge Infrastructure) from 2004 to 2013. Search key words were "产后抑郁症", "Postpartum Depression", and we selected 27 studies except for non-clinical studies, insufficient number of studies, unrelated studies and tests on animals. Results: We selected 27 articles with following results: 1) One study of Jadad Quality Assessment Scale score showed 3 points, 4 studies showed 2 points and 22 studies showed less than 1 point. Overall, the quality of the studies was low. 2) DSM - IV was frequently used as a diagnostic criteria and HAMD was mostly used as evaluation criteria. Diagnostic criteria and evaluation criteria were the same in HAMD and EPDS. 3) In TCM, there are various ways, such as herbal medicine, acupuncture, electro acupuncture, auricular acupuncture and/or massage to treat the postpartum depression. 4) The roots of Bupleurum falcatum Linne, Glycyrrhiza uralensis Fisch, Angelica gigas Nakai and Paeonia lactiflora Pall, the rhizome of Atractylodis macrocephala Koidz, the semen of Zizyphus jujuba Mill, the radix of Curcuma aromatica Salisb, the sclerotium of Poria cocos Wolf and the rhizomas of Cyperus rotundus L and of Cnidium officinale Makino are mainly used in the herbal medicine. BaekHoi (GV20) and Yindang (Ex-HN3) are mainly used in acupuncture points. 5) Most of the studies showed effective results. Some of the results were similar to those of the control groups which were cured by western medicine. Other results were better than those of control groups. Conclusions: Research on postpartum depression has been actively conducted in TCM and all studies showed effective results in the study group patients. Therefore, with this study, we hope to activate more clinical research on the treatment of patients in Korean traditional medicine.

• Journal of Acupuncture Research
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• 제22권1호
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• pp.223-234
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• 2005

우울증(憂鬱症)의 철구치료(鐵灸治療)에 관(關)한 중의학(中醫學) 문헌(文獻)을 조사(調査)하여 다음과 같은 결론(結論)을 얻었다. 1. 우울증(憂鬱症)을 통치(通治)하는 혈(穴)에서 다용(多用)된 혈(穴)은 내관(內關), 신문(神門), 족삼리(足三里), 백회(百會), 태충(太衝), 삼음교(三陰交)였다. 2. 우울증(憂鬱症)을 실증(實證)과 허증(虛證)으로 구분했을 때 다용(多用) 된 혈(穴)은 책증(責證)에서는 태충(太衝), 전중, 양릉천(陽陵泉), 풍륭(豊隆), 내관(內關)이었고, 허증(虛證)에서는 내관(內關), 삼음교(三陰交), 신문(神門), 심유(心兪)였다. 실증(實證)에는 사법(瀉法), 허증(虛證)에는 보법(補法)을 사용(使用)하였다. 3. 우울증(憂鬱症)에 다용(多用)된 혈(穴)은 영심안신(寧心安神), 소간해울(疏肝解鬱), 건장화위(健將和胃), 관흉화담(寬胸化痰) 등(等)의 특성(特性)이 있다. 4. 이철료법(耳鐵療法)에서는 다용(多用)된 혈(穴)은 신문(神門), 심(心), 침(枕), 피질하(皮質下)였고, 전침료법(電鍼療法)에서는 족삼리(足三里), 삼음교(三陰交), 신문(神門), 내관(內關), 통리(通里), 용천혈(涌泉穴) 등(等)이 사용(使用)되었으며, 혈위주사료법(穴位注射療法)에서는 심유(心兪), 전유, 족삼리(足三里)가 다용(多用) 되었다. 5. 피부침(皮膚鍼)은 주로 항배부(項背部) 독맥(督脈)과 방광경위주(膀胱經爲主) 혈위(穴位)에 피부(皮膚)가 홍윤(紅潤)해질 정도로 고자(叩刺)하는 방법(方法) 을 사용(使用)하였다. 6. 우울증(憂鬱症)의 치료에는 약물(藥物), 침구치료(鍼灸治療) 외에 정신요법(精神療法), 음악요법(音樂療法), 광선용법(光線療法) 등이 응용(應用)될 수 있다고 사려(思慮)된다.

• Clinical and Experimental Reproductive Medicine
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• 제31권3호
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• pp.155-168
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• 2004

Objective: Melatonin, which is secreted by pineal gland play an important role in the regulation of ovarian function via seasonal rhythm and sleep in most mammals. It also has a role in the protection of cells by removing toxic oxygen free radicals brought about by metabolism. In the present study, effects of melatonin on the mouse oocyte maturation were examined using two different culture conditions provided with 5% or 21% oxygen concentration. Material and Method: Immature mouse oocytes were obtained from the ovarian follicles of $3{\sim}4$ weeks old ICR strain mice intraperitoneally injected with 5 I.U. PMSG 44 hour before. Under stereomicroscope, morphologically healthy oocytes with distinct germinal vesicle (GV) were liberated from the graafian follicles and collected using mouth-controlled micropipette. They were then cultured for 17 hour at $37^{circ}C$, 5% $CO_2$ and 21% $O_2$ (95% air) or 5% $CO_2$, 5% $O_2$ and 90% $N_2$. New modified Hank's balanced salt solution (New MHBS) was used as a culture medium throughout the experiments. Effects of melatonin were examined at a concentration of $0.0001{\mu}M$, $0.01{\mu}M$ or $1.0{\mu}M$. For the prevention of spontaneous maturation of immature oocytes during culture, dibutyryl cyclic AMP (dbcAMP) and/or hypoxanthine were included in the medium. Results: Under 21% oxygen condition, oocytes cultured in the presence of $0.01{\mu}M$ melatonin showed a significantly higher maturation rates, in terms of germinal vesicle breakdown (95.0% vs 89.0%) and polar body formation (88.1% vs 75.4%), compared to those cultured with $0.0001{\mu}M$ or $1.0{\mu}M$ melatonin. However, no difference was observed in oocytes cultured under 5% oxygen whether they were treated with melatonin or not. In the presence of $0.01{\mu}M$ melatonin, oocytes either cultured under 21% or 5% oxygen exhibited no difference in the polar body formation (85.6% vs 86.7%). However, in the absence of melatonin, oocytes cultured under 21% oxygen exhibited lower polar body formation (74.7%). When oocytes were cultured in the presence of dbcAMP alone or with varying concentrations of melatonin, those treated with both compounds always showed better maturation, i.e., germinal vesicle breakdown and polar body formation, compared to those cultured with dbcAMP alone. At the same concentration of melatonin, however, oocytes exposed to 21% oxygen showed poor maturation than those to 5% oxygen. Similar results were obtained from the experiments using hypoxanthine instead of dbcAMP. Conclusion: Based upon these results, it is suggested that melatonin could enhance the meiotic maturation of mouse oocytes under 21% oxygen concentration, and release oocytes from the meiotic arrest by dbcAMP or hypoxanthine regardless of the concentration of oxygen, probably via the removal of oxygen free radicals.

• 한국수정란이식학회지
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• 제32권3호
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• pp.201-207
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• 2017

소 난자의 체외 성숙 및 발달과정에서 항산화제의 첨가는 발생과정에 생성될 수 있는 ROS를 조절하여 체외발생에 도움을 주는 것으로 알려져 있으나 이에 대한 연구는 아직 미흡하다고 판단된다. 본 연구에서는 소 수정란의 성숙과정과 발생과정에서 ROS에 대한 방어 기작에 필요한 물질로 -SH기(thiol group)을 함유하고 있는 NAC, NACA, GSH 및 CYS를 첨가하여 COCs의 난자 성숙율과 체외 수정 후 수정란의 발생율을 조사하였다. 도축장 유래 난소의 성숙율은 항산화제 처리군과 대조군에서 차이를 보여주지 않았으나(p>0.05), 배반포 형성율은 0.1 mM CYS을 처리한 실험군에서 $32.3{\pm}5.0%$로 유의적으로 높게 관찰되었다(p<0.05). 항산화제 0.3 mM NAC, 0.2 mM NACA 또는 0.5 mM GSH를 처리하는 실험군에서 배반포 형성율은 각각 $18.8{\pm}3.7%$, $21.2{\pm}3.9%$ 및 $26.5{\pm}5.0%$로 조사되었다. 그러므로, 항산화 물질인 NAC, NACA, GSH 및 CYS을 난자의 성숙 및 수정란 배양과정에 첨가하면 난자의 성숙에 영향이 없으나, CYS처리군이 배반포형성율에 효과가 있음을 밝혔다(p<0.05).

• Korean Journal of Acupuncture
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• 제40권3호
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• pp.79-89
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• 2023

Objectives : The purpose of this study is to investigate various application methods of focused ultrasound and apply them to clinical use in Korean medicine. Methods : Search was performed using the search engines of electronic databases, including PubMed, ScienceDirect, Cumulative Index to Nursing and Allied Health Literature (CINAHL), ScienceON, Oriental Medicine Advanced Searching Integrated System (OASIS), China National Knowledge Infrastructure (CNKI), Wanfang Data, Japan Science Technology Information Aggregator, Electronic (J-STAGE) and Citation Information by NII (CiNii), from inception to July 2023 without language limitation. Inclusion criteria were clinical studies including randomized controlled trials (RCTs), and animal experimental studies related with focused ultrasound treatments for acupoints or meridian sinews. Results : Total 17 papers, 7 for RCT, 6 for in vivo animal studies, and other experimental studies, were finally selected. Indications used in studies were shoulder pain, back pain, chronic back pain, and degenerative knee arthritis. In experimental studies, studies on animal models of hypoxic ischemic brain damage and hyperlipidemia were also conducted. As for the acupoints, LR3, LI4, and ST36 were used in clinical studies and, in animal experimental studies, GV20, KI1, and ST36 were used. As for the dose, 4 studies below 3 W/cm² and 3 studies in the range of 0.625 to 5 W/cm² in clinical studies, and all studies did not exceed 5 W/cm². In animal experimental studies, 0.5 W/cm², 2 W/cm², 7.5 WW/cm², 15 W/cm², 10~20 W/cm² were used. In all three studies describing the penetration depth during irradiation, it was less than 1 cm. Conclusions : We suggest that focused ultrasound is an appropriate treatment tool for stimulating the acupoints to transfer heat energy. Future studies with rigorous and well-designed RCTs for various diseases will be required to ascertain the focused ultrasound stimulate acupoints or meridian sinews.

• 한국수정란이식학회지
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• 제19권3호
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• pp.219-227
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• 2004

본 연구는 체외 성숙액인 TCM-199 배지를 기초로 무혈청첨가물인 PVA, PVP 및 pFF의 한정배지를 이용하여 돼지 미성숙난자의 체외성숙, 수정 및 배양 후 난자의 수정율과 배발달에 미치는 영향과 나아가 BSA 대체물로서의 이용 가능성을 알아보았다. 1. 무혈청 첨가물을 이용하여 난포란의 체외 성숙을 유기한 결과 PVA, PVP, pFF, BSA의 전체 분할율은 각각 82.4%, 78.6%, 89.4%, 90.0%로 나타났으며, GV, MI-MII율은 PVA 첨가시 각각 15.1%, 84.9%, PVP는 각각 26.5%, 73.5%, pFF는 각각 11.8%, 88.2%, BSA는 각각 11.1%, 88.9%로 PVA 혹은 pFF 첨가시에는 모두 BSA와 유사한 결과를 보였으나 PVP와는 유의적인 차이를 나타내었다 (P<0.05). 2. 체외 성숙된 난자를 수정시킨 후의 배발달율을 확인한 결과 전체 난할율은 PVA가 73%, PVP는 64.1% , pFF가 77.2%, BSA가 73%로 PVA와 pFF는 BSA와 유사하게 나타났으나, PVP는 그 발달율이 다른 처리군 들과 비교하여 유의적(P<0.05)으로 낮은 결과를 보였다. 3. 각 처리구간의 morulae와 blastocyst의 합을 비교한 결과 체외성숙 첨가물 중 PVA와 pFF는 각각 63%, 69%로 BSA(65%)와 유사한 결과를 나타내었으나 PVP는 54%로 유의적으로 낮은 결과를 나타내었다(P<0.05). 4. 전체 성숙율과 수정율을 비교해 본 결과 성숙 율은 PVA, pFF, BSA가 82.4%, 89.4%, 90.0%인 반면 PVP는 72.4%로 유의적으로(P<0.05) 낮은 결과를 보였고, 수정율은 pFF, BSA가 각각 87.1%, 89.1%로 PVA, PVP의 78.0%, 70.6%에 비해 각각 유의적(P<0.05)으로 높았다. 이상의 연구결과 돼지 난자의 체외성숙 및 배양 배지에 있어 PVA, pFF 첨가는 BSA 대체물로서 이용이 가능하나 PVP는 그 이용에 제한이 따를 것으로 판단된다.

• Clinical and Experimental Reproductive Medicine
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• 제36권3호
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• pp.175-186
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• 2009

목 적: Hypoxanthine (Hx)과 FSH가 미성숙난자의 배양에 미치는 영향을 관찰하기 위해 미성숙난자를 배양하여 GVBD, MII기 발생률을 비교 관찰하였다. 연구방법: 단순배양액인 BSAL-XI-HTF 배양액을 사용하여 (1) 0.3% BSA mBASAL-XI-HTF (0.3% BSA 배양액), (2) 0.1 IU/ml FSH를 첨가한 0.3% BSA mBASAL-XI-HTF (FSH 0.3% BSA 배양액), (3) 10% FBS mBASAL-XI-HTF (10% FBS 배양액), (4) 0.1 IU/ml FSH를 첨가한 10% FBS mBASAL-XI-HTF (FSH 10% FBS 배양액)의 4종류의 배양액을 만들었고, 각 종류의 배양액에서 생쥐 난구세포로 둘러싸인 미성숙난자의 성숙을 3, 6, 18시간별로 비교 관찰하였다. 각 배양액에 1 mM, 2 mM, 4 mM 농도의 미성숙난자성숙억제제인 Hx을 섞어 난자의 성숙억제양상을 관찰하였고 GVBD와 MII기 발생률을 비교 관찰하였다. 결 과: Hx을 첨가하지 않은 4종류의 배양액에서 미성숙난자의 자연성숙은 3시간 내에 대부분 GVBD가 발생하였고 MII기로의 발육은 6시간 이후 발생하였다. 18시간 후 각 군의 배양에서 모두 유사한 GVBD 발생률을 보였다. 1 mM, 2 mM, 4 mM Hx 농도의 0.3% BSA 배양액과 10% FBS 배양액에서 난자의 성숙억제양상을 비교해 보면 4 mM 농도의 Hx 배양액에서 18시간 동안 완전한 성숙억제를 보였다. 2 mM 농도의 Hx 배양액에서도 18시간 배양까지 억제를 보였으나 4 mM 농도의 Hx 배양액보다 작은 난자성숙억제양상을 보였다. 1 mM 농도의 Hx 배양액에서는 모두 난자성숙억제를 보이지 못했다. FSH를 첨가한 배양액에서는 2 mM 농도의 Hx 뿐만 아니라 1 mM 농도의 Hx 첨가에서도 초기 3시간까지 GVBD 성숙이 억제되었다. 또 같은 농도의 Hx을 함유하고 FSH를 첨가하지 않은 배양액에 비해 FSH를 첨가한 배양액에서 3시간, 6시간 동안 GVBD 발생을 더 억제하였다. 그러나 18시간 배양 후 난자성숙이 회복됨을 보였다. 이 결과로 FSH가 배양초기에 난자성숙을 억제하나 후기에는 성숙을 촉진함을 알 수 있었다. 18시간 후 MII기발육은 4 mM 농도의 Hx을 함유한 모든 배양액에서 낮은 발생률을 보였고, FSH를 함유한 10% FBS 배양액에 1 mM, 2 mM의 낮은 농도의 Hx을 첨가한 배양액은 다른 배양액에 비해 통계학적으로 높은 MII기 발생률을 보였다. 결 론: 본 실험에서 낮은 농도의 Hx과 FSH는 FBS를 함유한 배양액에서 미성숙난자의 초기배양 동안 성숙을 억제한 후 MII기 성숙을 촉진함을 알 수 있었다.

• Korean Journal of Acupuncture
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• 제29권4호
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• pp.519-536
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• 2012

Objectives : The purpose of this study is to figure out which acupoints are selected to treat low back pain and attempt to analyze the effectiveness of those acupoints in current clinical trials. Methods : We searched the three electronic databases(PUBMED, RISS, KISS) and manually checked related Korean journals and reference lists up to April 2012. We included randomized controlled trials, clinical controlled trials, and case reports/series using needle type acupuncture( manual acupuncture, electronic acupuncture) to treat low back pain in English and Korean. We investigated the frequency of selected acupoints, change of visual analogue scale of low back pain and statistical significance in each study among trials. Results : We included 37 articles(Domestic 17, International 20) 53 studies(Domestic 28, International 25) in this study. The most frequently adopted acupoints were BL23, BL24, BL25, GB30, BL26, BL60, BL32, BL40, KI3, GV3, ST36 in domestic studies and BL23, BL25, BL40, BL60, GB30, GB34, BL32, BL26 in international studies. There were differential effectiveness of acupoints between domestic and the international studies using statistical significance of visual analog scale for low back pain. Conclusions : These results suggest that both proximal and distal acupoints based on meridian theory were used in clinical trials to treat low back pain. It would be helpful to provide clinical guideline, evaluate the results of clinical trials appropriately, and reveal the effectiveness of acupoints.