• Title/Summary/Keyword: in-vitro

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Plasma Activity of Lysosomal Enzymes in Active Pulmonary Tuberculosis (활동성 폐결핵 환자에서 혈중 리소솜 효소의 활성도)

  • Koh, Youn-Suck;Choi, Jeong-Eun;Kim, Mi-Kyung;Lim, Chae-Man;Kim, Woo-Sung;Chi, Hyun-Sook;Kim, Won-Dong
    • Tuberculosis and Respiratory Diseases
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    • v.42 no.5
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    • pp.646-653
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    • 1995
  • Background: The confirmative diagnosis of pulmonary tuberculosis(Tb) can be made by the isolation of Mycobacterium Tuberculosis(MTb) in the culture of the sputum, respiratory secretions or tissues of the patients, but positive result could not always be obtained in pulmonary Tb cases. Although there are many indirect ways of the diagnosis of Tb, clinicians still experience the difficulty in the diagnosis of Tb because each method has its own limitation. Therefore development of a new diagnostic tool is clinically urgent. It was reported that silica cause some lysosomal enzymes to be released from macrophages in vitro and one of these enzymes is elevated in workers exposed to silica dust and in silicotic subjects. In pulmonary Tb, alveolar macrophages are known to be activated after ingestion of MTb. Activated macrophages can kill MTb through oxygen free radical species and digestive enzymes of lysosome. But if macrophages allow the bacilli to grow intracellularly, the macrophages will die finally and local lesion will enlarge. Then it is assumed that the lysosomal enzymes would be released from the dead macrophages. The goal of this investigation was to determine if there are differences in the plasma activities of lysosomal enzymes, ($\beta$-glucuronidase(GLU) and $\beta$-N-acetyl glucosaminidase(NAG), among the groups of active and inactive pulmonary Tb and healthy control, and to see if there is any possibility that the plasma activity of GLU and NAG can be used as diagnostic indicies of active pulmonary Tb. Methods: The plasma were obtained from 20 patients with bacteriologically proven active pulmonary Tb, 15 persons with inactive Tb and 20 normal controls. In 10 patients with active pulmonary Tb, serial samples after 2 months of anti-Tb medications were obtained. Plasma GLU and NAG activities were measured by the fluorometric methods using 4-methylumbelliferyl substrates. All data are expressed as the mean $\pm$ the standard error of the mean. Results: The activites of GLU and NAG in plasma of the patients with active Tb were $21.52{\pm}3.01$ and $325.4{\pm}23.37$(nmol product/h/ml of plasma), respectively. Those of inactive pulmonary Tb were $24.87{\pm}3.78$, $362.36{\pm}33.92$ and those of healthy control were $25.45{\pm}4.05$, $324.44{\pm}28.66$(nmol product/h/ml of plasma), respectively. There were no significant differences in the plasma activities of both enzymes among 3 groups. The plasma activities of GLU at 2 months after anti-Tb medications were increased($42.18{\pm}5.94$ nmol product/h/ml of plasma) in the patients with active pulmonary Tb compared with that at the diagnosis of Tb(P-value <0.05). Conclusion: The results of the present investigation suggest that the measurement of the plasma activities of GLU and NAG in the patients with active pulmonary Tb could not be a useful method for the diagnosis of active Tb. Further investigation is necessary to define the reasons why the plasma activities of the GLU was increased in the patients with active pulmonary Tb after Tb therapy.

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Comparison of Serum Cytokines($IL-1{\beta}$, IL-6, and $TNF-{\alpha}$) between Terminal Cancer Patients Treated with Vitamin C and Them without Vitamin C Therapy (Anorexia-Cachexia Syndrome을 가진 말기 암 환자에서 비타민 C 사용여부에 따른 사이토카인 변화 비교)

  • Yeom, Chang-Hwan;Suh, Sang-Youn;Cho, Kyung-Hee;Sun, Young-Gyu;Park, Yong-Gyu;Lee, Hye-Ree
    • Journal of Hospice and Palliative Care
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    • v.6 no.1
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    • pp.51-57
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    • 2003
  • Purpose : Anorexia-cachexia syndrome is one of the most common symptoms and main cause of death in terminal cancer patients. This symptom is due to the enlarged cancer mass as well as tumor released cytokines. Some doctors have suggested that vitamin C was preferentially toxic to tumor cells in vitro and in vivo, and improved clinical symptoms in terminal cancer patients. Therefore, we measured cytokines in serum of terminal cancer patients to determine whether vitamin C treatment improved the anorexia-cachexia syndrome. Methods : We investigated that 49 terminal cancer patients admitted to the department of family medicine, National Health Insurance Corporation Ilsan hospital from March 1, 2002 to August 31, 2002. The study was done on 22 patients who were given 10 g/day of vitamin C infusions during 1 week and 27 patients who were not infused. We measured the cytokines levels ($IL-1{\beta}$, IL-6, and $TNF-{\alpha}$) before and after 1 week between terminal cancer patients treated vitamin C and without vitamin C. Results : Out of 49 patients, patents treated with vitamin C infusions were 22 (12 male, 10 female), and these without vitamin C were 27 (18 male, 9 female). In patients treated with vitamin C, $IL-1{\beta}\;were\;6.19{\pm}5.17$ before day and $8.76{\pm}5.72$ after 1 week, IL-6 were $3.07{\pm}8.09$ before day and $1.31{\pm}2.36$ after 1 week, and $TNF-{\alpha}\;were\;2.74{\pm}14.24$ before day and $0.50{\pm}2.00$ after 1 week. In patients treated without vitamin C, $IL-1{\beta}\;were\;2.50{\pm}3.58$ before day and $6.49{\pm}12.01$ after 1 week, IL-6 were $1.00{\pm}2.19$ before day and $17.16{\pm}81.55$ after 1 week, and $TNF-{\alpha}\;were\;1.19{\pm}2.98$ before day and $1.27{\pm}1.52$ after 1 week. The level of cytokines in patients treated with vitamin C decreased more than those without vitamin C. However, this represented no statistical value (P=0.0598 in $IL-1{\beta}$, P=0.1664 in IL-6, and P=0.5395 in $TNF-{\alpha}$). Conclusion : In terminal cancer, even if there was no statistical difference in the cytokines levels between patients treated with vitamin C and those not treated, those who were treated had a decrease all cytokines levels. Vitamin C is very safe with almost no side effects. Therefore, vitamin C treatment in terminal cancer patients can be seen as beneficial and helpful for clinical symptoms.

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Studies on Garlic Mosaic Virus -lts isolation, symptom expression in test plants, physical properties, purification, serology and electron microscopy- (마늘 모자이크 바이러스에 관한 연구 -마늘 모자이크 바이러스의 분리, 검정식물상의 반응, 물리적성질, 순화, 혈청반응 및 전자현미경적관찰-)

  • La Yong-Joon
    • Korean journal of applied entomology
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    • v.12 no.3
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    • pp.93-107
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    • 1973
  • Garlic (Allium sativum L.) is an important vegetable crop for the Korean people and has long been cultivated extensively in Korea. More recently it has gained importance as a source of certain pharmaceuticals. This additional use has also contributed to the increasing demand for Korean garlic. Garlic has been propagated vegetatively for a long time without control measures against virus diseases. As a result it is presumed that most of the garlic varieties in Korea may have degenerated. The production of virus-free plants offers the most feasible way to control the virus diseases of garlic. However, little is known about garlic viruses both domestically and in foreign countries. More basic information regarding garlic viruses is needed before a sound approach to the control of these diseases can be developed. Currently garlic mosaic disease is most prevalent in plantings throughout Korea and is considered to be the most important disease of garlic in Korea. Because of this importance, studies were initiated to isolate and characterize the garlic mosaic virus. Symptom expression in test plants, physical properties, purification, serological reaction and morphological characteristics of the garlic mosaic virus were determined. Results of these studies are summarized as follows. 1. Surveys made throughout the important garlic growing areas in Korea during 1970-1972 revealed that most of the garlic plants were heavily infected with mosaic disease. 2. A strain of garlic mosaic virus was obtained from infected garlic leaves and transmitted mechanically to Chenopodium amaranticolor by single lesion isolation technique. 3. The symptom expression of this garlic mosaic virus isolate was examined on 26 species of test plants. Among these, Chenopodium amaranticolor, C. quince, C. album and C. koreanse expressed chlorotic local lesions on inoculated leaves 11-12 days after mechanical inoculation with infective sap. The remaining 22 species showed no symptoms and no virus was recovered from them whet back-inoculated to C. amaranticolor. 4. Among the four species of Chtnopodium mentioned above, C. amaranticolor and C. quinoa appear to be the most suitable local lesion test plants for garlic mosaic virus. 5. Cloves and top·sets originating from mosaic infected garlic plants were $100\%$ infected with the same virus. Consequently the garlic mosaic virus is successively transmitted through infected cloves and top-sets. 6. Garlic mosaic virus was mechanically transmitted to C, amaranticolor when inoculations were made with infective sap of cloves and top-sets. 7. Physical properties of the garlic mosaic virus as determined by inoculation onto C. amaranticolor were as follows. Thermal inactivation point: $65-70^{\circ}C$, Dilution end poiut: $10^-2-10^-3$, Aging in vitro: 2 days. 8. Electron microscopic examination of the garlic mosaic virus revealed long rod shaped particles measuring 1200-1250mu. 9. Garlic mosaic virus was purified from leaf materials of C. amaranticolor by using two cycles of differential centrifugation followed by Sephadex gel filtration. 10. Garlic mosaic virus was successfully detected from infected garlic cloves and top-sets by a serological microprecipitin test. 11 Serological tests of 150 garlic cloves and 30 top-sets collected randomly from seperated plants throughout five different garlic growing regions in Korea revealed $100\%$ infection with garlic mosaic virus. Accordingly it is concluded that most of the garlic cloves and top-sets now being used for propagation in Korea are carriers of the garlic mosaic virus. 12. Serological studies revealed that the garlic mosaic virus is not related with potato viruses X, Y, S and M. 13. Because of the difficulty in securing mosaic virus-free garlic plants, direct inoculation with isolated virus to the garlic plants was not accomplished. Results of the present study, however, indicate that the virus isolate used here is the causal virus of the garlic mosaic disease in Korea.

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A Report on the Shigella Cultures Isolated in Korea(1971) (1971년(年) 한국(韓國)에서 분리(分離)한 이질균(痢疾菌)에 관(關)한 보고(報告))

  • Ryu, Young-Hat;Kim, Soon-Hee
    • The Journal of the Korean Society for Microbiology
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    • v.7 no.1
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    • pp.9-15
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    • 1972
  • The authors identified 45 Shigella cultures among 63 suspectable cultures obtained from the cases showing dysentery or the like symptoms in various parts of the country during the period from February to September, 1971. Of 45 cultures, 36 cultures belonged to Subgroup B, 7 cultures to Subgroup C and 2 cultures to Subgroup D. There was none of cultures belonging to Subgroup A in 1971, although the authors detected one culture of that among 41 identified Shigella in 1967. Of 36 cultures belonging to Subgroup B, 27 cultures were $B_{2a}$ and 3 cultures of each $B_{3c},\;B_{4a}$ and $B_y$ were serotyped, respectively. Of 7 cultures beloning to Subgroup C, one was $C_{11}$ and other six cultures were $C_{15}$ and all S. boydii were isolated in Seoul area. It might be possible to suggest that there was a tendency of decreasing prevalence of S. dysenteriae and of increasing prevalence of S. boydii in Korea, although the number of cultures. tested were very much limited to conclude, if it was compared with the results obtained by the authors in 1967 that the fourty-one identified Shigella were composed of one culture of S. dysenteriae, 36 cultures of S. flexneri, 4 cultures of S. sonnei and none of S. boydii. According to the results obtained from the biochemical tests, the positive ratio of Indol tests in Subgroup B was 31/36, which could be higher than 9/36 observed in 1961, but which was nearly correlated with the result published by Ewing and his colleagues in U.S.A. The positive rates of both sorbitol and raffinose were lower than that observed by Ewing and his colleagues in S. flexneri. Regarding with the sensitivity of Shigella cultures to the antibiotics being widely used in the country, 46.2% were sensitive to chloramphenicol, which was very much higher than that observed by Park, and 85.8% were sensitive to ampicillin, which would be the drug of choice according to the results from the In Vitro tests.

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Effect of Early Pregnant Heifer as Donor on the Ovum Pick-Up Derived Oocyte Aspiration and Embryo Production (초기 임신우의 공란우 활용이 초음파 유도 난자 채취 및 수정란 생산에 미치는 영향)

  • Jin, Jong-In;Kwon, Tae-Hyeon;Choi, Byeong-Hyun;Kim, Sung-Soo;Jo, Hyun-Tea;Bang, Jae-Il;Kim, Sam-Chul;Cho, Kyu-Woan;Lee, Jung-Gyu;Kong, Il-Keun
    • Journal of Embryo Transfer
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    • v.26 no.1
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    • pp.19-25
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    • 2011
  • This study was carried out to evaluate the effect of early pregnant cow as donor for Ovum Pick-Up (OPU) derived oocyte aspiration and embryo production in Holstein heifers. Four non-pregnant and 2 pregnant Holstein heifers were used as donor and then carried out total 17 OPU session for 10 weeks (2 times per week). Recovered cumulus-oocyte-complexes (COCs) were classified into 4 grade by oocyte cytoplasm and cumulus cells and matured in vitro in TCM-199 supplemented with 10% FBS, 10 mg/ml FSH and 1 mg/ml estradiol in 5% $CO_2$ and over 99% humidity for 24 h. After 24 h co-incubation with post-thaw sperm, the presumed zygotes were cultured in CR1aa medium with 4 mg/ml BSA for 3 days and then changed CR1aa medium with 10% of FBS for another 3~4 days. The Mean number of aspirated follicles and collected oocytes in the early stage pregnant and non-pregnant heifers were $13.0{\pm}4.3$ and $10.6{\pm}3.9$, $5.4{\pm}3.4$ and $7.7{\pm}3.6$ per session, respectively. Rate of collected oocyte from aspirated follicles were 59.2% and 50.5%, respectively. The average number of good quality oocytes (Grade I and II) in the early stage pregnant and non-pregnant heifers was $3.7{\pm}2.7$ and $4.9{\pm}2.6$ (Mean${\pm}$SD). Cleavage and blastocyst developmental rates in Grade I and II were 22.2% and 25.5%, and then $1.7{\pm}0.9$ and $1.4{\pm}1.1$ blastocyst per session, respectively. In conclusion, OPU technology can be used in early stage pregnant and non-pregnant heifers without any problem and so applied OPU derived embryo production to maximize the ability of genetically valuable females.

Factors Affecting Pregnancy Rates on Transfer of Pronuclear Microinjected Embryos in Korean Black Goats (전핵 미세 주입법으로 생산된 한국흑염소 수정란의 이식 조건이 수태율에 미치는 영향)

  • Choi, Y.S.;Shin, H.G.;Jang, S.K.;Yang, H.S.;Lee, O.K.;Lee, D.S.;Cho, J.K.;Shin, S.T.
    • Journal of Embryo Transfer
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    • v.22 no.1
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    • pp.53-61
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    • 2007
  • This study was investigated factors affecting the pregnancy rates after transfer of pronuclear microinjected embryos for the production of transgenic Korean black goats. Embryo transfer was carried out in 343 recipient Korean black goats from September 1999 to June 2000. Estrus was induced by the insertion of intravaginal progesterone devices $CIDR^(R)$ for 2 weeks. A single injection of 400 IU equine chorionic gonadotropin was administered at 48h before $CIDR^(R)$ removal to increase the proportion of does cycling and ovulation rate. Good quality embryos were prepared by microinjection of DNA into the pronuclei of fertilized goat oocyte and cultured in vitro. Pronuclear microinjected $1{\sim}8$ cell stage embryos were surgically transferred into the oviducts of the recipient at day 4 or 5 following $CIDR^(R)$ removal, and morula to blastocyst stage embryos were surgically transferred into uterus at day 9. Pregnancy was diagnosed by transrectal ultrasound scanning at $20{\sim}30d$ and 8 weeks following embryo transfer. The pregnancy rate was affected by several factors, such as estrus induction, the number of previous transfer, transfer site, stage of CL (corpus luteum), the number of recipient CL, stage of embryos and the number of transferred embryo. The pregnancy rate was significantly higher in recipients that came into estrus naturally than recipients that induced to come into estrus with $CIDR^(R)$(59.1% vs. 36.8%; P<0.05). The pregnancy rate was higher when the embryos were transferred into the left oviduct than transferred into the right oviduct (42.9% vs. 35.3%; P<0.05). The pregnancy rate of recipients with $CH_1$ (early) stage corpus hemorrhagicum in ovary was hi틴or than recipient with $CH_3$ (late) stage hemorrhagicum (47.5% vs. 17.9%; P<0.01). Higher pregnancy rates were obtained by transfer of 1-cell stage embryos into oviduct while late blastocysts (51.6% vs. 66.7%; P<0.01) into uterus. The pregnancy rates when 3 embryos were transferred to recipients were significantly higher than when 2 embryos we.e transferred (47.6% vs. 27.0%; P<0.05). Although there were no significant difference among the group, adhesion of reproductive organs, uterine size, ovulation rate of recipients, presence of large follicle and difficulty of transfer affected pregnancy rate of recipient. Higher pregnancy rates were obtained in the recipients with $8{\sim}15m$ diameter uterine horn as compared to the recipients with <5m diameter or >20mm diameter uterine hem (38.9%, 20% vs. 18.2%), in the recipients with large follicle in the ovulated ovary ipsilaterally (53.6% vs. 37.1%) and in the transfer which was carried out easily (39.2% vs. 27.8%, 0%). In conclusion, the high rate of pregnancy was achieved following transfer of pronuclear microinjected embryos when three or four 1-cell stage embryos were transferred into oviduct with $CH_1$ stage corpus hemorrhagicum in the ovary of recipient which came into estrus naturally.

A Phase I/II Trial of $DCVac/IR^{(R)}$ Dendritic Cell Immunotherapy Combined with Irradiation in Cases of Refractory Colorectal Cancer with Multiple Liver Metastases (기존의 치료에 반응하지 않는 다발성 간전이 대장암 환자에서 방사선조사와 병합한 수지상세포 면역치료의 1, 2상 임상시험)

  • Choi, Young-Min;Lee, Hyung-Sik;Kwon, Hyuk-Chan;Han, Sang-Young;Choi, Jong-Cheol;Chung, Ju-Seop;Kim, Chang-Won;Kim, Dong-Won;Kang, Chi-Duk
    • Radiation Oncology Journal
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    • v.26 no.2
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    • pp.104-112
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    • 2008
  • Purpose: To assess the toxicity and tumor response induced by $DCVac/IR^{(R)}$ dendritic cell(DC) immunotherapy combined with irradiation for refractory colorectal cancer patients with multiple liver metastases. Materials and Methods: Between May 2004 and November 2006, applicants from a pool of refractory colorectal cancer patients with multiple liver metastases were enrolled. The patients were registered after having signed the informed consent form, which had been approved by the Institutional Review Board from the Dong-A University and Busan National University Hospital. DCs were obtained from peripheral blood of each patient, and then cultured in vitro. A total of $6{\times}10^6$ DCs were packed into a vial($DCVac/IR^{(R)}$, 0.5 ml) at the convenience of each patient's schedule. On the day before and on the day of each vaccination, each patient received a 4 Gy radiation dose to the target tumor. On the day of vaccination, the indicated dose of autologous DCs was injected into the irradiated tumor using ultrasound-guided needle injection procedures. A total of four vaccinations were scheduled at three 2-week intervals and one 4 week interval at the Dong-A University and Busan National University Hospital. If the tumor status was deemed to be stable or responding to therapy, an additional vaccination dose or two was approved at 4 week intervals beyond the fourth immunization. A tolerance test for DCs was conducted by injecting a range of doses($3{\times}10^6\;to\;12{\times}10^6$ DCs) after the 3rd injection. Moreover, the maximal tolerable dose was applied to additional patients. Treatment safety was evaluated in all patients who had at least one injection. Treatment feasibility was evaluated by the 10th week by assessing the response of patients having at least 4 injections. For systemic toxicities, the evaluation was performed using the National Cancer Institute Common Toxicity Criteria, whereas adverse effects were recorded using common WHO toxicity criteria. Results: Of the 24 registered patients, 22 received the DCs injections. Moreover, of the 14 patients that applied for the tolerance test, only 11 patients completed it because 3 patients withdrew their testing agreement. A grade 3 or more side effect, which was possibly related to the DC injection, did not occur in additional patients. The $12{\times}10^6$ DC injection was identified as the maximum tolerable dose, and was then injected in an additional 8 patients. Patients tolerated the injection fairly well, with no fatal side effects. In order to assess the feasibility of DC immunotherapy, the response was evaluated in other hepatic lesions outside of the targeted hepatic lesion. The response evaluation was performed in 15 of the 17 patients who received at least 4 injections. Stable and progressive disease was found in 4 and 11 patients, respectively. Conclusion: The DC-based immunotherapy and radiotherapy is theoretically synergistic for the local control and systemic control. The $DCVac/IR^{(R)}$ immunotherapy combined with irradiation was tolerable and safe in the evaluated cases of refractory colorectal cancer with multiple liver metastases. Future work should include well designed a phase II clinical trials.

ART Outcomes in WHO Class I Anovulation: A Case-control Study (저성선자극호르몬 성선저하증 여성에서 보조생식술의 임신율)

  • Han, Ae-Ra;Park, Chan-Woo;Cha, Sun-Wha;Kim, Hye-Ok;Yang, Kwang-Moon;Kim, Jin-Young;Koong, Mi-Kyoung;Kang, Inn-Soo;Song, In-Ok
    • Clinical and Experimental Reproductive Medicine
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    • v.37 no.1
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    • pp.49-56
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    • 2010
  • Objective: To investigate assisted reproductive technology (ART) outcomes in women with WHO class I anovulation compared with control group. Design: Retrospective case-control study. Methods: Twenty-three infertile women with hypogonadotropic hypogonadism (H-H) who undertook ART procedure from August 2003 to January 2009 were enrolled in this study. A total of 59 cycles (H-H group) were included; Intra-uterine insemination with super-ovulation (SO-IUI, 32 cycles), in vitro fertilization with fresh embryo transfer (IVF-ET, 18 cycles) and subsequent frozenthawed embryo transfer (FET, 9 cycles). Age and BMI matched 146 cycles of infertile women were collected as control group; 64 cycles of unexplained infertile women for SO-IUI and 54 cycles of IVF-ET and 28 cycles of FET with tubal factor. We compared ART and pregnancy outcomes such as clinical pregnancy rate (CPR), clinical abortion rate (CAR), and live birth rate (LBR) between the two groups. Results: There was no difference in the mean age ($32.7{\pm}3.3$ vs. $32.6{\pm}2.7$ yrs) and BMI ($21.0{\pm}3.1$ vs. $20.8{\pm}3.1kg/m^2$) between two groups. Mean levels of basal LH, FSH, and $E_2$ in H-H group were $0.62{\pm}0.35$ mIU/ml, $2.60{\pm}2.30$ mIU/ml and $10.1{\pm}8.2$ pg/ml, respectively. For ovarian stimulation, H-H group needed higher total amount of gonadotropin injected and longer duration for ovarian stimulation (p<0.001). In SO-IUI cycles, there was no significant difference of CPR, CAR, and LBR between the two groups. In IVF-ET treatment, H-H group presented higher mean $E_2$ level on hCG day ($3104.8{\pm}1020.2$ pg/ml vs. $1878.3{\pm}1197.7$ pg/ml, p<0.001) with lower CPR (16.7 vs. 37.0%, p=0.11) and LBR (5.6 vs. 33.3%, p=0.02) and higher CAR (66.7 vs. 10.0%, p=0.02) compared with the control group. However, subsequent FET cycles showed no significant difference of CPR, CAR, and LBR between the two groups. Conclusion: H-H patients need higher dosage of gonadotropin and longer duration for ovarian stimulation compared with the control groups. Significantly poor pregnancy outcomes in IVF-ET cycles of H-H group may be due to detrimental endometrial factors caused by higher $E_2$ level and the absence of previous hormonal exposure on endometrium.

A Study of Guidelines for Genetic Counseling in Preimplantation Genetic Diagnosis (PGD) (착상전 유전진단을 위한 유전상담 현황과 지침개발을 위한 기초 연구)

  • Kim, Min-Jee;Lee, Hyoung-Song;Kang, Inn-Soo;Jeong, Seon-Yong;Kim, Hyon-J.
    • Journal of Genetic Medicine
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    • v.7 no.2
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    • pp.125-132
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    • 2010
  • Purpose: Preimplantation genetic diagnosis (PGD), also known as embryo screening, is a pre-pregnancy technique used to identify genetic defects in embryos created through in vitro fertilization. PGD is considered a means of prenatal diagnosis of genetic abnormalities. PGD is used when one or both genetic parents has a known genetic abnormality; testing is performed on an embryo to determine if it also carries the genetic abnormality. The main advantage of PGD is the avoidance of selective pregnancy termination as it imparts a high likelihood that the baby will be free of the disease under consideration. The application of PGD to genetic practices, reproductive medicine, and genetic counseling is becoming the key component of fertility practice because of the need to develop a custom PGD design for each couple. Materials and Methods: In this study, a survey on the contents of genetic counseling in PGD was carried out via direct contact or e-mail with the patients and specialists who had experienced PGD during the three months from February to April 2010. Results: A total of 91 persons including 60 patients, 49 of whom had a chromosomal disorder and 11 of whom had a single gene disorder, and 31 PGD specialists responded to the survey. Analysis of the survey results revealed that all respondents were well aware of the importance of genetic counseling in all steps of PGD including planning, operation, and follow-up. The patient group responded that the possibility of unexpected results (51.7%), genetic risk assessment and recurrence risk (46.7%), the reproduction options (46.7%), the procedure and limitation of PGD (43.3%) and the information of PGD technology (35.0%) should be included as a genetic counseling information. In detail, 51.7% of patients wanted to be counseled for the possibility of unexpected results and the recurrence risk, while 46.7% wanted to know their reproduction options (46.7%). Approximately 96.7% of specialists replied that a non-M.D. genetic counselor is necessary for effective and systematic genetic counseling in PGD because it is difficult for physicians to offer satisfying information to patients due to lack of counseling time and specific knowledge of the disorders. Conclusions: The information from the survey provides important insight into the overall present situation of genetic counseling for PGD in Korea. The survey results demonstrated that there is a general awareness that genetic counseling is essential for PGD, suggesting that appropriate genetic counseling may play a important role in the success of PGD. The establishment of genetic counseling guidelines for PGD may contribute to better planning and management strategies for PGD.

The Mechanism of Interferon-$\gamma$ Induced Cytotoxicity on the Lung Cancer Cell Line, A549 (인터페론감마에 의한 A549 폐암세포주 세포독성의 기전)

  • Oh, Yeon-Mok;Yoo, Chul-Gyu;Chung, Hee-Soon;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.1
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    • pp.63-68
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    • 1996
  • Background: Interferon-$\gamma$ has various biologic effects, including antiviral effect, antitumor proliferative effect, activation of macrophage and B lymphocyte, and increased expression of major histocompatibility complex. Especially, antitumor proliferative effect of interferon-$\gamma$ has already been proved to be important in vivo as well as in vitro. And, clinical studies of interferon-$\gamma$ have been tried in lung cancer patients. However, the mechanism of antitumor effect of interferon-$\gamma$ has not yet been established despite of many hypotheses. "Necrosis" is a type of cell death which is well known to occur in the circumstances of severe stresses. In contrast, "apoptosis" is another type of cell death which occurs in such biological circumstances as embryonic development, regression of organs, and self-tolerance of lymphocytes. And, apoptosis is an active process of cell death in which cells are dying with fragmentations of their cytoplasms and nuclei. And, in the process of apoptosis the DNAs of cells are cleaved between nucleosomes by unidentified endonuclease and therefore DNAs of apoptotic cells result in a typical electrophoresis pattern known as DNA ladder pattern. Recently it has been suggested that cytotoxic effect of interferon-$\gamma$ occurs via apoptosis. To elucidate the mechanism of antitumor cytotoxic effect of interferon-$\gamma$, we microscopically observed a lung cancer cell line, A549 which was treated with interferon-$\gamma$. We observed A545 treated with interferon-$\gamma$ was dying fragmented. And so, we performed this study to find out that the mechanism of antitumor cytotoxic effect of interferon-$\gamma$ be apoptosis. Method: We treated A549, human lung cancer cell line with various concentration of interferon-$\gamma$ and quantified its cytotoxic effect of various periods, 24 hours, 72 hours and, 120 hours by MTT(dimethylthiazolyl diphenyltetrazolium bromide) bioassay. Also, after we treated A549 with 100 units/mi of interferon-$\gamma$ for 120 hours, we observed the pattern of cell death with inverted microscope and we extracted DNAs from the dead A549 cells and observed the pattern of 1.5% agarose gel electrophoresis with ethidium bromide staining. Result: 1) Cytotoxic effect of interferon-$\gamma$ on A549: For the first 24 hours, threre was little cytotoxic effect and for between 24 hours and 72 hours, there was the beginning of cytotoxic effect and for 120 hours there was increased cytotoxic effect. 2) Pattern of A549 cell death by interferon-$\gamma$: We observed with inverted microscope that A549 cells were dying fragmented. 3) DNA ladder pattern of gel electrophoresis: We observed DNA ladder pattern of gel electrophoresis of extracted DNAs from dead A549 cells. Conclusion: We concluded that the mechanism of interferon-$\gamma$induced cytotoxicity on lung cancer cell line, A549 be via apoptosis.

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