• Journal of Embryo Transfer
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• v.20 no.1
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• pp.63-69
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• 2005

This study were carried out to evaluate the possibility of nuclear progression of canine immature oocytes, of which was cultured in a reproductive tract, such as oviduct, ovarian bursa and uterus of estrus bitch for 4, 5 and 6 days following immediately collection. Cumulus intact oocytes(COC) fore collected from domestic dog following ovariohysterectomy at local veterinary clinics. In Exp. 1, COCs $of>110\;{\mu}m$ diameter were selected and cultured in vitro at $39^{\circ}C$, $5\%\;CO\_{2} $ in air atmosphere. The nuclear progression of canine oocytes checked at 24, 48 or 72 h after in vitro maturation. There was not increased the nuclear progression to the M II stage depending on culture periods at 24, 48 and 72h $(1.3\%,\;3.7\%\;and\;4.7\%)$. In Exp. 2, to evaluate of nuclear progression of immature oocytes, collected or in vitro cultured oocytes were transfer into a canine reproductive tract (oviduct, ovarian bursa and uterus). The recovery rates of canine oocytes from a reproductive tract after 4 days $(33.7\%)$ in vivo culture were significantly higher than those 5 $(17.7\%)$ 6 day $(3.4\%)$ (P<0.05). The survival rates of collected oocytes after 4 days $(60.0\%)$ were also significantly higher than those of 5 days $(30.2\%)$ and 6 days $(38.9\%)$ (P<0.05). The meiotic resumption rates of canine oocytes were not significantly difference among the culture periods at 4 days $(5.9\%)$, 5 days $(0.0\%)$ and 6 days $(0.0\%)$. These results show that the nuclear progression of canine immature oocytes from in in vivo culture was not affect the nuclear resumption of oocytes.

• Journal of Embryo Transfer
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• v.25 no.3
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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.

• Journal of Acupuncture Research
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• v.22 no.1
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• pp.223-234
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• 2005

Objective : The objective of this study was to research depression with acupuncture & moxibustion treatment. Methods : We searched Chinese Medical Literature(published from 1985~2002) related to depression and acupuncture. Results : 1) PC6(內關), HT7(神門), ST36(足三里), GV20(百會), LR3(太衝), SP6(三陰交) are much used for the treatment depression. 2. LR3(太衝), CV17(전중), GB34(陽陵泉), ST40(豊隆), PC6(內關) are much used for the excess type of depression and PC6(內關), SP6(三陰交), HT7(神門), BL15(心兪) are much used for deficiency type of depression. 3. ST36(足三里), SP6(三稜交), HT7(神門), PC6(內關), HT5(通里), KI1(通泉) are used for the treatment depression in electroacupuncture therapy. BL15(心兪), BL20(脾兪),ST36(足三里) are used for the treatment depression in acupoint injection therapy. 4. In dermal needle therapy, it is used to tap slightly until the skin becomes congested in the region of Bladder meridian and nape, back of Governor Vessel. 5. It seems that psychotherapy, music therapy and light therapy will be applied for the treatment of depression. Conclusion : We expect that acupuncture treatment of depression will be applied practically in clinical medicine due to further study on depression with acupuncture.

• Korean Journal of Acupuncture
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• v.22 no.1
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• pp.33-41
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• 2005

Objectives : This study was to investigate the effect of the Carthami Semen Herbal acupuncture therapy on a Cauda equina syndrome patient who has a complex of low back pain, bilateral sciatica, saddle anesthesia and motor weakness in the lower extremity and paraplegia with bladder and bowel incontinence. Methods : Oriental Medical Therapy was performed on the Cauda equina syndrome patient from July 15th 2004 to July 29th 2004. The patient was treated with Carthami Semen Herbal acupuncture at BL22, BL23, BL25, BL28 and GV3 in combination with herbal medicine and conventional body acupuncture. We evaluated The Visual Analog Scale(VAS), Improvement index, The Oswestry Diability Index(ODI), gaiting, dyschezia, bladder incontinence, duration of urination and area of anesthesia, Digital Infrared Thermographic Imaging(DITI) before and after treatment. Results : 1. After treatment, VAS, Improvement index, ODI were improved each from 10 to 2, from 21 to 73, from 333 to 166. 2. After treatment, gaiting, bladder incontinence, duration of urination and area of anesthesia and DITI were improved well, but dyschezia was remained. Conclusions : From this case it is thought Carthami Semen herbal acupuncture therapy is very effective to Cauda Equina Syndrome and further study is needed for the confirmation of the effect of Carthami Semen Herbal acupuncture.

• Korean Journal of Animal Reproduction
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• v.14 no.1
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• pp.1-8
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• 1990

These experiments were carried out to investigate the effects of cumulus cells for in vitro fertilization and development of bovine follicular oocytes matured in vitro. The bovine ovaries were obtained at a slaughter house and the follicular oocytes surrounded by cumulus cells were collected by puncturing follicles with 2~6 mm of diameter. Bovine oocytes were matured in vitro for 24~26 hours in a CO2 incubator with 5% CO2 in air at 39$^{\circ}C$. The medium used for maturation was TCM-199 supplemented with hormones, pyruvate, FCS and antibiotics. Epididymal spermatozoa were capacitated by in vitro culture for 2~3 hours in BO solution containing BSA(5mg/ml) and caffeine(2.5mM). Insemination was made by introducing about 10~15 matured oocytes into the suspension of capacitated spermatozoa. Six hour after insemination the eggs were transferred to TCM-199 supplemented with FCS(10%) and HEPES(25mM), cultured for 7~8 days with 10~15 eggs/well in 4-well multidishes(Nunc Co.) forming cumulus cell monolayer. The results obtained in these experiments were summarized as follows ; 1. The majority of the follicular oocytes with compacted cumulus cells existed in GV stage while those with dispersed or denuded cumulus cells existed GVBD and M II stage. 2. After 24~26 hours maturation, the maturation rates of the follicular oocytes cultured in TCM-199 containing hormones were slightly higher than those of oocytes cultured in medium without hormones, and the frequency of cumulus compacted or denuded oocytes reaching M II stage cultured in medium containing hormones was 75.7% or 51.7%, respectively(P<0.05). 3. After 20 hours in vitro insemination, percentages of ova fertilized were 61.4% or 51.4%, respectively, for cumulus oophorus intacted or removed, and increased frequency of ova with both male and female pronuclei was found when cumuli were present(P<0.05). 4. The rates of embryos developed to 2-, 4-, 8-, 16-cell and morula or blastocyst stage after cocultured with cumulus cells were 65.0%, 45.3%, 34.7%, 28.0% and 22.7%, respectively. The results for momla or blastocyst stage were significantly higher than those of the embryos cultured in the basic medium(P<0.05).

• Clinical and Experimental Reproductive Medicine
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• v.32 no.4
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• pp.331-336
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• 2005

Objective: To investigate the effects of female age on in vitro maturation and fertilization of immature oocytes from controlled ovarian hyperstimulation (COH) in human IVF-ET program. Method: A total of 96 immature oocytes (GV & metaphase I) obtained from 40 cycles of IVF-ET (29 patients). The mean age of female patients was $31.8{\pm}3.1years$. Ovulation was triggered by urinary or recombinant hCG. Immature oocytes were cultured with YS medium containing 30% of patients' human follicular fluids, LH (1 IU/mL), FSH (1 IU/mL) and EGF (10 ng/mL), and then matured oocytes were fertilized by ICSI. In vitro maturation and fertilization of immature oocytes were analyzed according to age of female (< 34 or ${\geq}34years$). Results: The maturation rate was similar between two groups (68% vs 64%). The fertilization rate of in?vitro-matured oocytes was higher in patients < 34 years old, but there was no statistical significance (64% vs 50%, p=0.347). The fertilization rate of in-vitro-matured oocytes was significantly lower compared with those of in-vivo-matured oocytes in both age groups (64% vs 79%, p=0.035, 50% vs 86%, p=0.007). Conclusion: In older female group, fertilization rate of in-vitro-matured oocytes seems to be decreased. Further investigations should be warranted to increase fertilization potential of in-vitro-matured oocytes.

• Clinical and Experimental Reproductive Medicine
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• v.31 no.3
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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.

• Journal of Embryo Transfer
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• v.32 no.3
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• pp.201-207
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• 2017

To increase the productivity of in vitro development, the antioxidants have been used for culture system of bovine oocytes and embryos. However, comparative studies on these molecules are rare and direct beneficial effects on blastocyst production cannot be discriminated for best results. The study was conducted to determine the influence of N-acetyl-L-cysteine (NAC), N-acetyl-L-cysteine amide (NACA), glutathione (GSH) and cysteamime (CYS) on maturation competence of COCs from GV to MII stage and productivity of blastocyst formation during in vitro fertilization and culture. There was no difference among maturation rates of oocytes to metaphase II with polar body with antioxidants for any of the treatment groups (p>0.05). However, the significant improvement on the rate of blastocysts ($32.3{\pm}5.0%$) was found in 0.1 mM CYS treatment than 0.3 mM NAC, 0.2 mM NACA or 0.5mM GSH (p<0.05). The addition of NAC ($18.8{\pm}3.7%$) or NACA ($21.2{\pm}3.9%$) did not improve development competence to morula and blastocysts than control ($24.4{\pm}4.1%$) and GSH ($26.5{\pm}5.0%$) (p>0.05). Our study showed that medium supplementation with CYS during IVM and IVC improved the rate of bovine embryo development but not with NAC, NACA and GSH addition.

• Journal of Oriental Neuropsychiatry
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• v.25 no.2
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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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• v.32 no.3
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• pp.175-183
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• 2015

Objectives : The purpose of this study is to investigate the effects of Sumsu (Bufonis venenum) pharmacopuncture on insomnia and depression. Methods : We investigated 32 patients with a sleep disorder who had been recruited at a Korean medical hospital from March $1^{st}$, 2014 to April $16^{th}$, 2015. We divided subjects into two groups : The control group(N=15) was treated with Normal Saline and the experimental group(N=17) was treated with Sumsu(Bufonis venenum) pharmacopuncture. The subjects were injected with 0.05 ml~0.1 ml, for a total of 0.6 ml on each acupuncture point. These were Pungbu ($GV_{16}$), Pungi ($GB_{20}$), Gyeonjeong ($GB_{21}$), and Anmyun(EX-$HN_{20}$). Both groups were evaluated with the beck depression inventory(BDI) and given an insomnia score before treatment, after one week, two weeks and four weeks from the initial visit. Results : The Insomnia score and BDI decreased significantly in the experimental group. In comparing the control group with the experimental group, there was a more significant improvement of the insomnia score in the experimental group during all periods of this study. However, the difference of change in BDI between the control group and experimental group was only significant in the first week. Conclusions : Sumsu (Bufonis venenum) pharmacopuncture can be used as an effective treatment for patients with insomnia and depression.