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The Correction Factor of Sensitivity in Gamma Camera - Based on Whole Body Bone Scan Image - (감마카메라의 Sensitivity 보정 Factor에 관한 연구 - 전신 뼈 영상을 중심으로 -)

  • Jung, Eun-Mi;Jung, Woo-Young;Ryu, Jae-Kwang;Kim, Dong-Seok
    • The Korean Journal of Nuclear Medicine Technology
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    • v.12 no.3
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    • pp.208-213
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    • 2008
  • Purpose: Generally a whole body bone scan has been known as one of the most frequently executed exams in the nuclear medicine fields. Asan medical center, usually use various gamma camera systems - manufactured by PHILIPS (PRECEDENCE, BRIGHTVIEW), SIEMENS (ECAM, ECAM signature, ECAM plus, SYMBIA T2), GE (INFINIA) - to execute whole body scan. But, as we know, each camera's sensitivity is not same so it is hard to consistent diagnosis of patients. So our purpose is when we execute whole body bone scans, we exclude uncontrollable factors and try to correct controllable factors such as inherent sensitivity of gamma camera. In this study, we're going to measure each gamma camera's sensitivity and study about reasonable correction factors of whole body bone scan to follow up patient's condition using different gamma cameras. Materials and Methods: We used the $^{99m}Tc$ flood phantom, it recommend by IAEA recommendation based on general counts rate of a whole body scan and measured counts rates by the use of various gamma cameras - PRECEDENCE, BRIGHTVIEW, ECAM, ECAM signature, ECAM plus, IFINIA - in Asan medical center nuclear medicine department. For measuring sensitivity, all gamma camera equipped LEHR collimator (Low Energy High Resolution multi parallel Collimator) and the $^{99m}Tc$ gamma spectrum was adjusted around 15% window level, the photo peak was set to 140-kev and acquirded for 60 sec and 120 sec in all gamma cameras. In order to verify whether can apply calculated correction factors to whole body bone scan or not, we actually conducted the whole body bone scan to 27 patients and we compared it analyzed that results. Results: After experimenting using $^{99m}Tc$ flood phantom, sensitivity of ECAM plus was highest and other sensitivity order of all gamma camera is ECAM signature, SYMBIA T2, ECAM, BRIGHTVIEW, IFINIA, PRECEDENCE. And yield sensitivity correction factor show each gamma camera's relative sensitivity ratio by yielded based on ECAM's sensitivity. (ECAM plus 1.07, ECAM signature 1.05, SYMBIA T2 1.03, ECAM 1.00, BRIGHTVIEW 0.90, INFINIA 0.83, PRECEDENCE 0.72) When analyzing the correction factor yielded by $^{99m}Tc$ experiment and another correction factor yielded by whole body bone scan, it shows statistically insignificant value (p<0.05) in whole body bone scan diagnosis. Conclusion: In diagnosing the bone metastasis of patients undergoing cancer, whole body bone scan has been conducted as follow up tests due to its good points (high sensitivity, non invasive, easily conducted). But as a follow up study, it's hard to perform whole body bone scan continuously using same gamma camera. If we use same gamma camera to patients, we have to consider effectiveness of equipment's change by time elapsed. So we expect that applying sensitivity correction factor to patients who tested whole body bone scan regularly will add consistence in diagnosis of patients.

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Monitoring North Korea Nuclear Tests: Comparison of 1st and 2nd Tests (북한 핵실험 모니터링 : 1, 2차 비교)

  • Chi, Heon-Cheol;Park, Jung-Ho;Kim, Geun-Young;Che, Il-Young;Sheen, Dong-Hoon;Shin, Jin-Soo;Cho, Chang-Soo;Lee, Hee-Il
    • Geophysics and Geophysical Exploration
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    • v.13 no.3
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    • pp.243-248
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    • 2010
  • Two suspicious events, which were claimed as underground nuclear tests by North Korea, were detected in the northern Korean Peninsula on October 9, 2006 and May 25, 2009. The KIGAM and Korea-China Joint seismic stations are distributed uniformly along the boundaries between North Korea and adjacent countries. In this study, the data from broadband stations with the distance of 200 to 550 km from the test site are used to analyze and compare two nuclear tests of North Korea. By comparing the time differences of the Pn-wave arrival times of 1st and 2nd tests at multiple stations, the relative locations of two test sites could be calculated precisely. From the geometrical calculation with the velocity of Pn wave $V_{Pn}$ = 8 km/s, the 2nd test site is estimated to move in the WNW direction from 1st one with the distance of 2 km. Body wave magnitude, mb of the 2nd test, which was announced officially as the network average of 4.5, varies widely with the directional location of stations from 4.1 to 5.2. The magnitude obtained from Lg wave, $m_b$(Lg), shows less variation between 4.3 to 4.7 with the average of 4.6. The moving-window spectra of time traces of 1st and 2nd tests show very similar pattern with different scale level. In addition, the corner frequencies of P wave of 1st and 2nd tests at each station show no or negligible difference. This indicates the burial depths of two tests might be very similar. The relative yield amount of the 2nd test is estimated 8 times larger than that of the 1st from the weighted average of ground-velocity amplitude ratios.

현대여성(現代女性)의 의복의식(衣服意識)에 관한 조사(調査) 연구(硏究) - 서울 지역(地域)의 양복(洋服) 착용자(着用者)를 중심(中心)으로 -

  • Lee, Hee-Myung
    • Journal of the Korean Society of Costume
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    • v.2
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    • pp.73-88
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    • 1978
  • This article is an attempt to explain, at least in part, the contemporary Korean women's consciousness of Western Dreasses. As time changes, the role of clothing undergoes varisous transitions, while values and ways of life are constantly in change. It is, therefore, proper and appropriate to recognize as among the major aspects of social psychology such phenomenon as interests, understanding of clothing, the choice of a dress, and attitudes toward clothing, etc. The purpose of this study is to discover problems concerning and their clothing and their solutions, by means of a surveying approach. The method of research used is based upon questionares distributed to parents of first-year pupils in elementary schools and to female clerks working in offices, covering the period from August through October, 1976. The number of the questionares distrubuted totalled 600, and 526 were returned to the research to be utilized for analysis. The contents of the survey included such things as values concerning clothing, kinds of clothing and their practical use, the selection of clothing and the method of purchase, fashions, etc. The classification of aquisition are self-made clothing, clothing made to order and ready-made materials. It is composed of 25 items, including affirmative reasons as well as negative ones. The processing of the material returned was made by using the computer, and based upon classifications such as ages, monthly income, occupations; thus diagraming the result in percentages. The conclusion made and the improvements proposed are as follows: 1. The values of clothing were placed on the expression of the wearer's personality (32.7) and on eauty(28. 6%). The lower age group places is stress upon the expression of personality, while the higher age group stresses beauty. About 50% of wearers are contented with their clothing, their clothing, the rest of whom them indicating their dissatisfaction with what they wear. As to designs at the time of selection, about 46% indicated their preference of personal expression, 31.8% on usefulness. In selecting material, practicality is emphasized; in selecting patterns, single color is preferred. In short, personal expression and esthetic values are primary, with consideration of practicality in mind. 2. The classification of clothing according to their uses indicates the highest numbers in normal wear (home wears) and clothings to be worn outside home. As to evening dresses, (party dress) only one or two articles were checked by many, and no such article was clamed to be possessed by most. The highest ratio of wearing was shown in the case of home wear (47.3%) and clothing to be worn outside the home, which is 55.8%. The budget for one article of clothing was greatest in the case of home wear, and clothing worn outside the home. Many used both kinds of articles for the same purpose. It is desirable, therefore, that the kinds of clothing should be varied according to the purpose for which they are worn, and that clothing appropriate for that purpose should be worn. 3. The motivation for purchasing clothing was highly chosen in the item of seasonal change, which was 55.7%; Clothing deliberately made was indicated by 45.2%. In the mothods of purchasing clothing, clothing made to order and ready-made was indicated by 44.4%, which is the highest; Clothing made to order was 25.4%, and self-sewing was 1.1%, which is the lowest. (1) In the case of self-sewing, "I like it but it is very hard," was checked by 43.6%; "It is so difficult that I cannot wear such clothing" was checked by 13.3%. From these, we can conclude that the questionees are willing to make clothing by themselves, but techniques involved in sewing and at her problems involved in the skill are complicated but when those problems are eliminated there is a possibility for practice. The response checked by questionees concerning the self-sewing was, "It's economical", which is a clear indication that many questionees are positive for self-sewing. It is generally believed that ready-made clothing is cheaper, but it is not necessarily so. In consideration of the quality of clothing, self-sewing is a necessity, and it is desirable that it should be encouraged. (3) Problems involved in ready-made clothing, such as designs, skills, size (fitting) should be eliminated. When these problems are scientifically gotten rid of, it is possible that affirmative returns will be expected. Affirmative responses such as "Ready-made clothing is economical," "You can select there on the spot," are good signs that many women expect to wear ready-made clothing. It is in this sense that the prospect for ready-made clothing is brighter when much development for ready-made clothing is on the way. 4. Much concern for fashion are checked in such item of questions as "Fashionable clothing in the show window," "Clothes worn by women." The first item was checked by 50.1 %, and the second was checked by 48.6%. The reason for following fashion is "Because many people wear them," which was indicated by 30.4%. The reason for not following fashion is "It is too expensive," which was checked by 29.6%. The 26.2% of the answers indicated that "Fashionable clothing is devoid of personality," The influences of fashion over the development of fashion over the development of clothing are two-fold: Esthetic and active. It is not to be deniable that people follow fashion more or less. 1978.9>

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Flip Angle of the Optimal T1 Effect Using FLASH Pulse Sequence at 3T Abdominal MRI (FLASH를 이용한 3T 복부검사에 있어서 최적의 T1효과를 위한 적정 Flip Angle)

  • Han, Jae-Bok;Choi, Nam-Gil
    • Journal of radiological science and technology
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    • v.32 no.1
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    • pp.101-106
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    • 2009
  • Purpose of this study is to compare the signal intensity (SI) and CNR with T1 weighted image using FLASH at 3T abdominal MRI by varying flip angle (FA). Totally 20 patients (male : 12, female : 8, Age : $28{\sim}63$ years with mean : 51) were examined by 3 Tesla MR scanner (Magnetom Tim Trio, SIEMENS, Germany) with 8 channel body array coil between september and October 2008. Imaging parameters were as follows : FLASH sequence, TR : 120 ms, TE : minimum, FOV (field of view) : $360{\times}300\;mm$, Matrix : $256{\times}224$, slice : 6 mm, scan time : 15 sec and Breath-hold technique. Abdominal image, with a 50 ml syringe filled with water placed in the FOV measuring the water signal, were acquired with varying FA through $10^{\circ}$ to $90^{\circ}$ with $10^{\circ}$ interval. SI's were measured three times at liver parenchyme, water, spleen and background and averaged. The CNR's were measured between the ROIs (region of interest). Statistic analysis was performed with ANOVA test using SPSS software (version 17.0). Less than FA $30^{\circ}$, abdominal images were severely inhomogeneity. Especially, T1 effect of water signal was weak. As the flip angle increased, the signal intensity decreased at all the regions. Especially, flip angle of the highest signal intensity was observed with $40^{\circ}$ at the liver parenchyme, $20^{\circ}$ at water, $30^{\circ}$ at the spleen, respectively. The CNR between liver and water was -60.92 at FA $10^{\circ}$ and 15.16 at FA $80^{\circ}$. The CNR between liver and spleen was -3.18 at FA $10^{\circ}$ and 9.65 at $80^{\circ}$. In conclusion, FA $80^{\circ}$ is optimal for T1 weighted effect using FLASH pulse sequence at 3.0 T abdominal MRI.

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Development of Nutritional Counseling Internet Program for Weight Reduction Using Expert System (전문가 시스템을 이용한 인터넷 체중 감량 상담 프로그램 개발)

  • 박선민;박수진;최선숙
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.5
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    • pp.993-999
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    • 2001
  • The purpose of the study was to develop a nutritional counseling program using expert system to assist obese people to lose weight through behavior modification in the internet. The counseling internet program for weight loss was developed by the accumulation of knowledge for dealing with eating habits and exercising behaviors into expert system tool, Knowledge Engineering Agent (KEA) by a dietitian without any help of computer expert. KEA was built based on the theory of Multiple Classification Ripple Down Rules. To accumulate knowledge into KEA, survey was performed in 150 obese people, the dietitian reviewed and consulted each survey case, and the consulted contents were learned and accumulated into KEA. Survey questionnaires were the same as those of the internet consulting program, and they included general characteristics, dietary habits, lifestyle, and exercise patterns related to obesity. KEA was used for nutritional counseling of obese people after KEA had enough knowledge for weight loss based on behavior modification by the dietitian. To accumulate knowledge to KEA, the dietitian selected proper factors inferred from the survey questionnaire of each case, and added the conclusions for them. Conclusions were made for helping clients to correct bad eating behaviors and accumulate good behaviors for losing weight. When clients answered survey questionnaires in a counseling internet program, KEA gave the recommendation how to eat, to exercise and the deal with stress in a real time for each case. If KEA did not have enough knowledge for a specific case, the conclusion window wrote no conclusion and the dietitian needed to add conclusions for the case. The conclusions for the new case added to the KEA knowledge base. In conclusions, a counseling internet program for weight reduction can be used for give advices how to deal with obesity in a man-to-man way in a real time using KEA where nutritional knowledge based on behavior modification for weight loss was accumulated.

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Analysis of ethyl glucuronide (EtG) in Hair for the diagnosis of chronic alcohol abuse of Korean (한국인의 만성 알코올 중독 진단을 위한 모발에서 Ethyl Glucuronide (EtG) 분석법 연구)

  • Gong, Bokyoung;Jo, Young-Hoon;Ju, Soyeong;Min, Ji-Sook;Kwon, Mia
    • Analytical Science and Technology
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    • v.33 no.3
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    • pp.151-158
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    • 2020
  • Alcohol, which can easily be obtained in the same way as ordinary beverages, is harmful enough to cause death due to excessive drinking and chronic alcohol intake, so it is important to maintain a proper amount of drinking and healthy drinking habits. In addition, the incidence of behavioral disturbances and impaired judgments that can be caused by chronic alcohol drinking of more than adequate amounts of alcohol is also significant. Accordingly it is very useful for forensic science to check whether the person involved is drunken or is alcoholism state in various accidents. Currently, in Korea, alcohol consumption is determined by detecting the level of alcohol or alcohol metabolism 'ethyl glucuronide (EtG)' in blood or urine samples. However, analysis of alcohol or EtG in blood or urine can only provide information about the current state of alcohol consumption because of a narrow window of detection time. Therefore, it is important to analyze the EtG as a long-term direct alcohol metabolite bio-marker in human hair and to investigate relationship between alcohol consumption and EtG concentration for the evaluation of chronic ethanol consumption. In this study, we established an analytical method for the detection of EtG in Korean hair efficiently and validated selectivity, linearity, limits of detection (LOD), limits of quantification (LOQ), matrix effect, recovery, process efficiency, accuracy and precision using liquid chromatography tandem mass spectrometry (LC-MS/MS). In addition, the assay performance was evaluated in Korean social drinker's hair and the postmortem hair of a chronic alcoholism. The results of this study can be useful in monitoring the alcohol abuse of Korean in clinical cases and legal procedures related to custody and provide a useful tool to evaluate postmortem diagnosis of alcoholic ketoacidosis in forensics.

Effects of Maturation Duration and Activation Treatments on Activation and Development of Porcine Follicular Oocytes (돼지 난모세포의 단위발생에 있어서 성숙시간과 활성화 처리가 활성화와 발달에 미치는 영향)

  • Kim H. J.;Choi S. H.;Han M. H.;Son D. S.;Ryu I. S.;Kim I. C.;Lee J. H.;Kim I. H.;Im K. S.;Cho S. R.
    • Journal of Embryo Transfer
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    • v.20 no.1
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    • pp.25-33
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    • 2005
  • This study is a part of research that development of effective genetic resources preservation system using the in vitro spermatogenesis, in vitro insemination and culture system. We aimed for establishment of in vitro culture system with in vitro activated porcine oocytes. The porcine oocytes were matured for 48 hours in $TCM199+10\%$ FCS and activated with $7\%$ ethanol. The activated oocytes were cultured for 7 days in $TCM199+10\%$ FCS or $NCSU23+0.4\%$ BSA medium. The activated oocytes were not developed to the blastocyst stage in $TCM199+10\%$ FCS medium. However in $NCSU23+0.4\%$ medium, those were developed to blastocyst with $3\%$ of treated oocytes. We extended maturation duration of porcine follicular oocytes fur 48, 52, 56, 60, 64, 68, and 72 hours and activated with $7\%$ ethanol and cultured using $NCSU23+0.4\%$ BSA medium. The six percents of activated oocytes were developed to blastocyst in 48 hours and $10\%$ in 52 hours with comparatively low rates suggested to be not fully activated by regenerated MPF. Maturation durations from 56 hours to 68 hours supported to develop upto $11.9\~18.3\%$ of blastocysts. However the developmental rate was declined to $7.2\%$ at 72 hours of maturation duration because of cytoplasmic deterioration. The assumed time window for activation will be $56\~68$ hours of maturation duration. When the matured oocytes were activated with electric pulse of 1, 1.2, 1.4, 1.6, 1.8 and 2.0kV/cm for $80{\mu}s$, although appling the electric current once was not enough for activation, appling twice with 1.6kV/cm for $80{\mu}s$ was shown the highest developmental rate with $11.3\%$. When those were compared with activating methods, $15.7%$ of blastocyst rate was obtained in the $7\%$ ethanol. That was higher than those in electric pulse with $9.5\%$ and calcium ionophore method with $5.8\%$. In this experimental condition, the $7\%$ ethanol treatment was the most effective method for activating porcine oocytes.

One-stage Repair of Aortic Arch Anomalies and Intracardiac Defects through Median Sternotomy (정중 흉골절개를 통한 대동맥궁 기형과 심기형의 일차교정술)

  • Chang Yun Hee;Lee Sang Kwon;Lee Hyung Doo;Kim Siho;Yie Kilsoo;Woo Jong Soo;Lee Young Seok;Sung Si Chan
    • Journal of Chest Surgery
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    • v.38 no.4 s.249
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    • pp.291-300
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    • 2005
  • One-stage repair of aortic arch anomalies and intracardiac defects through median sternotomy has been recently adopted by many institutions since it is known to be safer than the staged repair. The early and midterm results of the one-stage repair of aortic arch anomalies and intracardiac defects were retrospectively evaluated. Material and Method: 45 patients who underwent one-stage repair of aortic arch anomalies and intracardiac defects performed by one surgeon from January 1996 to July 2003 were included in this retrospective study. The median age of repair was 16 days (range, 3 days$\~$23.7 months) and the mean weight was $3.62\;\pm\;1.30 kg$. Thirty one $(68.9\%)$ had coarctation and 14 $(31.1\%)$ had interrupted aortic arch. Associated intracardiac anomalies were VSD in 31 patients (VSD group), TGA or Taussig-Bing anomaly in 10 (TGA group), and others in 4 (ASD in 1, aortopulmonary window 1, truncus arteriosus 1, aortic and mitral stenoses 1, miscellaneous group). The arch obstruction was repaired with end-to-side anastomosis in 23 patients and end-to-end anastomosis in 22. Result: Overall postoperative hospital mortality was $22.2\%\;(10/45);\;16.1\%$ (5/31) in VSD group, $40\%$ (4/10) in TGA group, and$25\%$ (1/4) in miscellaneous group. There was no mortality in VSD group since 1998, and the mortality in TGA group has remarkably reduced since technical modification for coronary transfer was adopted $(75\%\;vs\;16.7\%)$. There was no postoperative seizure or other neurological complications. Postoperative aortic restenosis occurred in 5 patients $(5/35,\;14.3\%)$. Two patients underwent balloon aortoplasty with successful results. There was no reoperation. There was one late death caused by pneumonia 5 months after the operation. Two-year actuarial survival rate including operative death was $72.9\%$. Conclusion: The operative mortality of one-stage repair has been reduced with time and aortic restenosis rate was also acceptable. We concluded that this procedure is a reproducible procedure for aortic arch anomalies associated with intracardiac defects.

Neurochemical Profile Quantification of Regional Adult Mice Brain Using: ex vivo $^1H$ High-Resolution Magic Angle Spinning NMR Spectroscopy (생체 외 조직 고 분해능 Magic Angle Spinning을 이용한 정상 Adult Mice에서의 뇌 부위별 뇌 신경화학 대사물질 정량분석)

  • Lee, Do-Wan;Woo, Dong-Cheol;Lee, Sung-Ho;Kim, Sang-Young;Kim, Goo-Young;Rhim, Hyang-Shuk;Choi, Chi-Bong;Kim, Hwi-Yool;Lee, Chang-Wook;Choe, Bo-Young
    • Progress in Medical Physics
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    • v.21 no.1
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    • pp.35-41
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    • 2010
  • The purpose of this study is to quantitate regional neurochemical profile of regional normal adult mice brain and assess regional metabolic differences by using ex vivo $^1H$ high-resolution magic angle spinning nuclear magnetic resonance spectroscopy ($^1H$ HR-MAS NMRS). The animals were matched in sex and age. The collected brain tissue included frontal cortex, temporal cortex, thalamus, and hippocampus. Quantitative 1D spectra were acquired on 40 samples with the CPMG pulse sequence (8 kHz spectral window, TR/TE = 5500/2.2 ms, NEX = 128, scan time: 17 min 20 sec). The mass of brain tissue and $D_2O$+TSP solvent were 8~14 mg and 7~13 mg. A total of 16 metabolites were quantified as follow: Acet, NAA, NAAG, tCr, Cr, tCho, Cho, GPC + PC, mIns, Lac, GABA, Glu, Gln, Tau and Ala. As a results, Acet, Cho, NAA, NAAG and mIns were showed significantly different aspects on frontal cortex, hippocampus, temporal cortex and thalamus respectively. The present study demonstrated that absolute metabolite concentrations were significantly different among four brain regions of adult mice. Our finding might be helpful to investigate brain metabolism of neuro-disease in animal model.

Implantation Rate and Clinical Pregnancy Rate According to Dosage and Timing of Progesterone Administration for Secretory Endometrial Preparation in Frozen-Thawed Embryo Transfer Cycles (동결보존 배아이식에서 분비기 자궁내막 유도시 프로게스테론 투여 방법에 따른 착상율과 임신율의 비교)

  • Park, Chan-Woo;Hur, Kuol;Kim, Moon-Young;Song, Hyun-Jung;Kim, Hye-Ok;Yang, Kwang-Moon;Kim, Jin-Yeong;Song, In-Ok;Yoo, Keun-Jae;Cheon, Kang-Woo;Byun, Hye-Kyung;Koong, Mi-Kyoung;Kang, Inn-Soo
    • Clinical and Experimental Reproductive Medicine
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    • v.30 no.3
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    • pp.193-202
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    • 2003
  • Objective: To evaluate the difference of implantation rate (IR) and clinical pregnancy rate (CPR) between two protocols of endometrial preperation in women undergoing frozen-thawed embryo transfer (FET) cycles. Methods: This study was performed during the different time periods: A retrospective study from January 2000 to June 2001 (phase I) and a prospective study from July 2001 to March 2002 (phase II). All the patients received estradiol valerate (6 mg p.o. daily) starting from day 1 or 2 of the menstrual cycle without pituitary down regulation. Progesterone was administered around day 14 after sonographic confirmation of endometrial thickness $\geq$7 mm and no growing follicle. In Group A (n=88, 99 cycles) of phase I, progesterone was administered i.m. at a dose of 50 mg daily from one day prior to thawing of pronuclear (PN) stage frozen embryo or three days prior to thawing of 6-8 cell stage frozen embryo and then each stage embryos were trasnsferred 2 days or 1 day later after thawing. In Group B (n=246, 299 cycles) of phase I, patients recieved progesterone 100 mg i.m. from one day earlier than group A; two days prior to PN embryo thawing, four days prior to of 6-8 cell embryo thawing. During the phase II, to exclude any differences in embryo transfer procedures, in Group 1 (n=23, 28 cycles) of phase II embryo was transfered by one who have used the progesterone protocol since the phase I. In Group 2 (n=122, 139 cycles) of phase II embryo was transfered by one who use the progesterone protocol from the phase II. Results: When compared across the phase and group, there were no significant differences in the characteristics. During the phase I, there were significant increase in IR (14.4% vs 5.9%, p=0.001) and CPR (28.3% vs 14.5%, p=0.000) in group A. During the phases II, IR (11.8% vs 10.6%) and CPR (27.6% vs 27.3%) show no differences between two groups. Conclusions: In FET cycles, IR and CPR are increased significantly by the change of dosage and timing of progesterone administraton. And the timing is considered to be more important factor because the dosage of progesterone did not affect implantation window in previous studies. Therefore, we suggest that progesterone administration in FET cycle should begin from one day prior to PN stage embryo thawing and three days prior to 6-8 cell stage embryo thawing.