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Effects of FSH and LH on Maturation of Bovine Preantral Follicle

  • Kim, D. J.;H. J. Chung;S. J. Uhm;Lee, H. T.;K. S. Chung
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.30-30
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    • 2001
  • The culture of preantral follicles has important biotechnological implications through its potential to produce the large quantity of oocytes for embryo production, transgenesis research, conservation of rare breed, and a potential source of ovarian genetic material. The present study was conducted to establish the optimal conditions of in vitro culture for intact bovine preantral follicles; and to examine the developmental ability of oocytes derived from the in vitro-grown preantral follicles; and to investigate the effects of various concentrations of FSH and LH on these processes. Bovine preantral follicles (150 $\pm$ 1.2${\mu}{\textrm}{m}$), surrounded by theca cell, were isolated enzymetically and mechanically from ovarian cortical slides in Leibovitz L-15 medium containing 1 mg/$m\ell$ collagens and 0.2 mg/$m\ell$ DNase I and cultured for 25 days in the presence of different concentrations of bovine FSH and LH in $\alpha$MEM medium with insulin, transferrin, and selenite. The survival was tested by frypan Blue and Hematoxylin. The survival and growth rates of follicles were higher in FSH treatment groups than these in control (P<0.001), but there were no significant differences between the LH treatment groups and the control. In 25 days, the survival and growth rates of follicles in FSH and LH treatment group (50%, 300$\pm$1.0${\mu}{\textrm}{m}$) were higher than in FSH treatment group (40%, 244$\pm$0.5${\mu}{\textrm}{m}$) and the control group (25%, 160$\pm$ 1.0${\mu}{\textrm}{m}$). Fifty-five percent of healthy antral follicles were obtained, and 60% of the oocytes complete meiotic maturation to the metaphase II stage. Twenty-two percent of the mature oocytes underwent cleavage, and 9% developed to the blastocyst stage. In this study, in vitro-grown oocytes (111 $\pm$ $1.5mutextrm{m}$), under our culture conditions, were not equivalent in size to the in vivo-grown oocytes (130$\pm$1.3${\mu}{\textrm}{m}$). Therefore, these results suggest that bovine preantral follicles with intact theca cell can grow to the antral stage in 25days, and that oocytes from those follicles can acquire the meiotic competence and normally undergo fertilization and development to the blastocyst stage. However, the developmental capacity of in vitro-grown oocytes is presumably not comparable to those of the in vivo counterparts.

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Bone marrow stem cells incubated with ellipticine regenerate articular cartilage by attenuating inflammation and cartilage degradation in rabbit model

  • Mohammad Amjad Hossain;Soyeon Lim;Kiran D. Bhilare;Md Jahangir Alam;Baicheng Chen;Ajay Vijayakumar;Hakyoung Yoon;Chang Won Kang;Jong-Hoon Kim
    • Journal of Veterinary Science
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    • v.24 no.6
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    • pp.83.1-83.12
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    • 2023
  • Background: Ellipticine (Ellip.) was recently reported to have beneficial effects on the differentiation of adipose-derived stem cells into mature chondrocyte-like cells. On the other hand, no practical results have been derived from the transplantation of bone marrow stem cells (BMSCs) in a rabbit osteoarthritis (OA) model. Objectives: This study examined whether autologous BMSCs incubated with ellipticine (Ellip.+BMSCs) could regenerate articular cartilage in rabbit OA, a model similar to degenerative arthritis in human beings. Methods: A portion of rabbit articular cartilage was surgically removed, and Ellip.+BMSCs were transplanted into the lesion area. After two and four weeks of treatment, the serum levels of proinflammatory cytokines, i.e., tumor necrosis factor α (TNF-α) and prostaglandin E2 (PGE2), were analyzed, while macroscopic and micro-computed tomography (CT) evaluations were conducted to determine the intensity of cartilage degeneration. Furthermore, immuno-blotting was performed to evaluate the mitogen-activated protein kinases, PI3K/Akt, and nuclear factor-κB (NF-κB) signaling in rabbit OA models. Histological staining was used to confirm the change in the pattern of collagen and proteoglycan in the articular cartilage matrix. Results: The transplantation of Ellip.+BMSCs elicited a chondroprotective effect by reducing the inflammatory factors (TNF-α, PGE2) in a time-dependent manner. Macroscopic observations, micro-CT, and histological staining revealed articular cartilage regeneration with the downregulation of matrix-metallo proteinases (MMPs), preventing articular cartilage degradation. Furthermore, histological observations confirmed a significant boost in the production of chondrocytes, collagen, and proteoglycan compared to the control group. Western blotting data revealed the downregulation of the p38, PI3K-Akt, and NF-κB inflammatory pathways to attenuate inflammation. Conclusions: The transplantation of Ellip.+BMSCs normalized the OA condition by boosting the recovery of degenerated articular cartilage and inhibiting the catabolic signaling pathway.

The Results of Hyperfractionated Radiotherapy on Locally Advanced Non-Small Cell Lung Cancer (국소적으로 진행된 비소세포 폐암에 대한 과분할 방사선 치료의 성적)

  • Hur, Won-Joo;Lee, Hyung-Sik;Kim, Jeong-Ki;Choi, Young-Min;Lee, Ho-Jun;Youn-Seon-Min;Kim, Jae-seok;Kim, Hyo-Jin;Woo-Jong-Soo;Choi, Pill-Jo;Lee, Ki-Nam
    • Radiation Oncology Journal
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    • v.16 no.3
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    • pp.275-282
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    • 1998
  • Purpose : The effect of hyperfractionated radiotherapy on locally advanced non-small lung cancer was studied by a retrospective analysis. Materials & Methods : We analyzed sixty one patients of biopsy-confirmed, IIIA and IIIB non-small cell lung cancer. Using the ECOG performance scale, all the patients were scored less than 2. They were treated by curative hyperfractionated radiotherapy alone from Oct. 1992 to Oct. 1995 at the Department of Radiation Oncology. All the patients received 120cGy b.i.d with more than 6 hours interval between each fraction. The total dose of radiation was reached up to 6400-7080 cGy with a mean dose of 6934 cGy. The results were analyzed retrospectively. Results : The overall survival rate was 53 1$\%$ in 1 year, 9.9$\%$ in 2 years with a median survival time (MST) of 13.9 months. The progression free survival (PFS) rate was 37.0$\%$ in 1 year, 8.9$\%$ in 2 years. Twenty two Patients were classified as complete responders to this treatment and their MST was 19.5 months When this was compared with that of partial responders (MST: 11 7months), it was statistically significant (p=0.0003). Twenty nine patients of stage IIIA showed a better overall survival rate (1yr 63.3$\%$, 2yr 16.8$\%$) than IIIB patients (1yr 43.3$\%$, 2yr 3.6$\%$), which was also statistically significant (p=0.003). Patients with adenocarcinoma showed a better survival rate (1yr 64.3$\%$, 2yr 21.4$\%$) than that of squamous cell counterpart (1yr 49.4$\%$, 2yr 7.4$\%$), although this was not significant statistically (p=0.61). Two patients developed fatal radiation-induced pneumonia right after the completion of the treatment which progressed rapidly and they all died within 2 months. One patient developed radiation-induced fibrosis after 13 months. He refused further treatment and died soon after the development of fibrosis. Conclusion : Among locally advanced NSCLC, hyperfractionated radiotherapy was effective on stage IIIA patients by increasing MST with acceptable toxicities. Acute radiation-induced pneumonia should be carefully monitored and must be avoided during or after this treatment.

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Culture Conditions of Aspergillus oryzae in Dried Food-Waste and the Effects of Feeding the AO Ferments on Nutrients Availability in Chickens (건조한 남은 음식물을 이용한 Aspergillus oryzae균주 배양조건과 그 배양물 급여가 닭의 영양소 이용률에 미치는 영향)

  • Hwangbo J.;Hong E. C.;Lee B. S.;Bae H. D.;Kim W.;Nho W. G.;Kim J. H.;Kim I. H.
    • Korean Journal of Poultry Science
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    • v.32 no.4
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    • pp.291-300
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    • 2005
  • Two experiments were carried out to assess the appropriate incubation conditions namely; duration, moisture content and the ideal microbial inoculant for fermented dried food waste(EW) offered to broilers. The nutrient utilization of birds fed the FW diets at varying dietary inclusion rates was also compared with a control diet. In Experiment 1, different moisture contents(MC) of 30, 40, 50 and $60\%$ respectively were predetermined to establish the ideal duration of incubation and the microbial inoculant. A 1mL Aspergillus oryzae(AO) $(1.33\times10^5\;CFU/mL)$ was used as the seed inoculant in FW. This results indicated that the ideal MC for incubation was $40\~50\%$ while the normal incubation time was > 72 hours. Consequently, AO seeds at 0.25, 0.50, 0.75 and 1.00mL were inoculated in FW to determine its effect on AO count. The comparative AO count of FW incubated for 12 and 96 hours, respectively showed no significant differences among varying inoculant dosage rates. The FW inoculated with lower AO seeds at 0.10, 0.05 and 0.01mL were likewise incubated for 72 and 96 hours, respectively and no changes in AO count was detected(p<0.05). The above findings indicated that the incubation requirements for FW should be $%40\~50\%$ for 72 hours with an AO seed incoulant dosage rate of 0.10mL. Consequently, in Experiment II, after determining the appropriate processing condition for the FW, 20 five-week old male Hubbard strain were used in a digestibility experiment. The birds were divided into 4 groups with 5 pens(1 bird per pen). The dietary treatments were; Treatment 1 : Control(Basal diet), Treatment 2 : $60\%$ Basal+4$40\%$ FW, Treatment 3 : $60\%$ $Basal+20\%\;FW+20\%$ AFW(Aspergillus oryzae inoculate dried food-waste diet) and Treatment 4: $60\%$ Basal+$40\%$ Am. Digestibility of treatment 2 was lowed on common nutrients and amino acids compared with control(p<0.05) and on crude fat and phosphorus compared with AFW treatments(T3, T4)(plt;0.05). Digestibility of treatment 3 and 4 increased on crude fiber and crude ash compared treatment 2 (p<0.05). Digestibility of control was high on agrinine, leucine, and phenylalnine of essential amino acids compared with treatment 3 and 4(p<0.05), and diestibility of treatment 3 and 4 was improved on arginine, lysine, and threonine of essential amino acids. Finally, despite comparable nutrient utilization among treatments, birds fed the dietary treatment containing AO tended to superior nutrient digestion to those fed the $60\%$ Basa1+$40\%$ FW.

Linkage Map and Quantitative Trait Loci(QTL) on Pig Chromosome 6 (돼지 염색체 6번의 연관지도 및 양적형질 유전자좌위 탐색)

  • Lee, H.Y.;Choi, B.H.;Kim, T.H.;Park, E.W.;Yoon, D.H.;Lee, H.K.;Jeon, G.J.;Cheong, I.C.;Hong, K.C.
    • Journal of Animal Science and Technology
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    • v.45 no.6
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    • pp.939-948
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    • 2003
  • The objective of this study was to identify the quantitative traits loci(QTL) for economically important traits such as growth, carcass and meat quality on pig chromosome 6. A three generation resource population was constructed from cross between Korean native boars and Landrace sows. A total of 240 F$_2$ animals were produced using intercross between 10 boars and 31 sows of F$_1$ animals. Phenotypic data including body weight at 3 weeks, backfat thickness, muscle pH, shear force and crude protein level were collected from F$_2$ animals. Animals including grandparents(F$_0$), parents(F$_1$) and offspring(F$_2$) were genotyped for 29 microsatellite markers and PCR-RFLP marker on chromosome 6. The linkage analysis was performed using CRI-MAP software version 2.4(Green et al., 1990) with FIXED option to obtain the map distances. The total length of SSC6 linkage map estimated in this study was 169.3cM. The average distance between adjacent markers was 6.05cM. For mapping of QTL, we used F$_2$ QTL Analysis Servlet of QTL express, a web-based QTL mapping tool(http://qtl.cap.ed.ac.uk). Five QTLs were detected at 5% chromosome-wide level for body weight of 3 weeks of age, shear force, meat pH at 24 hours after slaughtering, backfat thickness and crude protein level on SSC6.

Report on the Effects Lipemic Specimen in Anti-ds DNA Antibody Test (Anti-ds DNA 항체 검사 시 Lipemic 검체의 영향에 관한 보고)

  • Cheon, Jun Hong;Kim, Whe Jung;Kim, Sung Ho;Moon, Hyoung Ho;Yoo, Seon Hee
    • The Korean Journal of Nuclear Medicine Technology
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    • v.18 no.1
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    • pp.153-157
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    • 2014
  • Purpose: SLE (systemic lupus erythematosus) is an inflammatory autoimmune disease, characterized by various autoantibody. The detection of Anti double-stranded DNA (Anti-ds DNA) is important in the diagnostics of SLE, and include the American College of Rheumatology (ACR) diagnostic criteria for SLE. Also SLE disease activity and correlativity with the level Anti-ds DNA antibody have been reported and Anti-ds DNA antibody quantitative test is very useful for tracing before and after SLE treatment. When These Anti-ds DNA antibody test (Farr assay: $^{125}I$ labeled ds-DNA and bound Anti-ds DNA antibodies complex in serum is precipitated by ammonium sulfate and used to centrifugation, measured it) inhaled supernatant after centrifugation, a lipemic specimen does not facilitate the formation of precipitate and also occurs situation was inhaled with precipitate. To solve these problems, The Influence of the degree of lipemic specimen was evaluated. Materials and Methods: September 2012 to February 2013, We selected lipemic samples (n=81) of specimen commissioned by Anti-ds DNA antibody test. Lipemic samples were done pre-treatment (high-speed centrifugation: 14,000 rpm 5 mins) used a micro-centrifuge (Eppendorf Model 5415D). At the same time lipemic specimen and pre-treatment samples were performed Anti-ds DNA antibody test (Anti-ds DNA kit, Trinity Biotech, Ireland). Statistical analysis were analyzed Pearson's correlation coefficients and regression and paired t-test, and Difference (%). Results: Experimental group 1 (Lipemic Specimen Anti-ds DNA Ab concentration ${\leq}7IU/mL$) at y=0.368X+4.732, $R^2=0.023$, Pearson's correlation coefficient was 0.154, paired t-test (P=0.003), Difference (%) mean 65.7 and showed a statistically significant difference. Experimental group 2 (Lipemic Specimen Anti-ds DNA Ab concentration ${\geq}8IU/mL$) at y=0.983X+0.298, $R^2=0.994$, Pearson's correlation coefficient showed 0.997, paired t-test (P=0.181), Difference (%) mean -5.53 made no statistically significant difference. Conclusion: Lipemic sample of low Anti-ds DNA Ab concentration (2.5-7 IU/mL) and the result is obtained pre-treatment (high-speed centrifugation: 14,000 rpm 5 mins) were made a significant difference statistically. Anti-ds DNA is one of the primary auto-antibodies present in patients with SLE, and remain an important diagnostic test for SLE. Therefore, we recommend preprocessing (high-speed centrifugation: 14,000 rpm 5 mins) in order to exclude the influence of lipemic specimen.

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Discrimination of Geographical Origin for Astragalus Root (Astragalus membranaceus) by Capillary Electrophoresis and Near-Infrared Spectroscopy (Capillary electrophoresis 및 근적외선분광분석기를 이용한 황기의 원산지 판별)

  • Kim, Eun-Young;Kim, Jung-Hyun;Lee, Nam-Yun;Kim, Soo-Jeong;Rhyu, Mee-Ra
    • Korean Journal of Food Science and Technology
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    • v.35 no.5
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    • pp.818-824
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    • 2003
  • Capillary electrophoresis (CE) and near-infrared spectroscopy (NIRS) were performed to discriminate astragalus roots (Astragalus membranaceus) according to geographical origin (domestic or foreign). Two-hundred-and-four astragalus roots were extracted with 30% methanol in 0.1 M phosphate buffer (pH 2.5) and separated in a uncoated fused-silica $(50\;{\mu}m{\times}27\;cm)$ capillary. Conditions for optimal analysis included: temperature $-45^{\circ}C$, voltage -14 kV, and pressure injection time -8 sec. The optimal separation buffer was 0.1 M phosphate buffer (pH 2.5) containing 40 mM hexane sulfonic acid with 20% 2-methoxy ethanol. Raw NIR spectra were obtained using NIRS, and modified partial least square regression was used to develop the prediction model. The correlation coefficient and standard error of prediction were 0.915 and 14.3%, respectively. Under the optimal conditions established for CE and NIRS, the geographical origins of the astragalus roots were correctly identified in 80 and 97%, respectively. Astragalus roots that were not discriminated by NIRS were correctly discriminated by CE. Hence, CE and NIRS are potential methods for discriminating the geographical origins of astragalus roots that complement one another.

Effects of Lime and NPK Application Rates on the Soil Charateristics after a 10-year Experiment in Oversown Hilly Pasture of Mixed Grass-Clover Sward III. Change in the mutural ratios of exchaegeable cations by the soil depth, and the visible characteristics of soil conservation (겉뿌림 산지초지에서 석회 및 3요소 시용수준이 10년후 토양특성에 미치는 영향 III. 토심별 염기간 상호비율 및 가시적 토양보존의 특성변화)

  • 정연규;이혁호
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.18 no.3
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    • pp.223-232
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    • 1998
  • The main experiment related to this report was undertaken to assess the effects of two rates of $Ca(OH)_2$ (0, 250 kg/10a only at establishment) and five rates of $N-P_20_5-K_20$ (0-0-0, 0-10-10, 6-15-15, 12-20-20, 24-25-20 kg/10a/year) on the pasture establishment, forage yield and quality, and vegetation etc. After this 10-year main e experiment pasture had been used to assess the effects of the above treaments on the soil characteristics in oversown hilly pasture of a grass-clover sward. soil properties of mutual ratios of exchangeable cations at d different soil depth, and the visible characteristics of soil conservation obtained are summarized as follows; 1. Compared with the properties of soil fertility and the level for the likelihood grass tetany, the mutual ratios of exchangeable cations in soils; Ca:Mg:K(% of CEC), Ca:Mg:K(K=I), $K/\sqrt{Ca+Mg}$, Mg/K, K/Mg, and Ca/Mg, were discussed at different soil depth. Before and after experiment, these ratios were generally unbalance and unsuitable, and were rather worsened in the order of Mg>Ca>K under the conditions of liming and NPK fertilization without Mg. 2. The ratios of Ca and Ca/Mg were increaby liming, whereas it of $K/\sqrt{Ca+Mg}$ was redused. The ratios of K and $K/\sqrt{Ca+Mg}$ in control and the heavy fertillization of NPK(especially N) were rather lowered than those in the low and medium fertilizations of NPK. 3. The prperties of consevation were closely related with the forage productivity/vegetation rates, as affected by liming and the application rates of NPK. In control of NPK, It was shown to be the worst soil conservation; severly eroded(3rd grade), exporsure of subsoil, redish brown in soil colour, and 18.8% of cobble and stony in covering rate. 4. The improvement of soil conservation was greatly enhanced by increasing the NPK rate. In the medium and heavy fertilizations of NPK, it was shown to be the favorable improvement of soil conservation; slightly eroded(lst grade), covered with humus layer/grass residues in surface soil, dark brown in surface soil colour, under 1% of cobble and stony in covering rate. The increasing of legume yield/vegetation rate by liming rather enhanced the soil conservation of grassland.

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Evaluation of Ventilation Systems in an Enclosed Nursery Pig House (무창자돈사의 환기시스템 정립 및 환기효율 평가)

  • Song, J.I.;Choi, H.L.
    • Journal of Animal Science and Technology
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    • v.44 no.1
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    • pp.123-134
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    • 2002
  • An experiment was conducted to establish the most suitable ventilation system for the enclosed nursery pig house in Korea, comparing four different ventilation systems ; i) air enters through perforated ceiling and exhausts through chimney (NA), ii) air enters through perforated ceiling and exhausts through side walls (NB), iii) air enters through perforated ducts and exhausts through side walls (NC) and iv) air enters through perforated ducts and exhausts through chimney(ND). The experiment was carried out during winter and summer separately. The experimental pigs were weaned at fourteen days old in winter (December-February) and at twenty one days old in summer (June-August). The main results of the experiment are as follows : A preliminary experiment showed that in the NC system during summer, air can reach all the pig rooms in the house and the air flow rates of the upper, middle (1.2 m height of the room) and low (at the height of pig stature) parts of the room were measured at 7.0-8.08, over 0.5 and over 0.2 m/s, respectively, which flow rates were much higher(p$<$0.05) than those in other system. At the minimum ventilation efficiency during winter, air flow rates of upper, middle and low parts of the room equipped with the NC system were detected at over 1, less than 0.5 and around 0.07 m/s, respectively. It is concluded that the separated ventilation system air-entering through ducts is the most suitable for the ventilation system of the enclosed nursery pig house and the exhausting system through side walls is more efficient for ventilation than the system through roof. Furthermore, to sustain proper temperature and reduce energy waste as well as heat consumption, a future research should be carried out to develop the environmental control system in relation to developing a heat regulator.

A STUDY ON THE PARENTAL MARITAL RELATIONSHIP OF CHILD PSYCHIATRIC PATIENTS (소아정신과 환아 부모의 부부관계에 대한 연구)

  • Lim, Ke-Won;Hong, Kang-E;Rhee, Kun-Hoo
    • Journal of the Korean Academy of Child and Adolescent Psychiatry
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    • v.2 no.1
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    • pp.160-175
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    • 1991
  • The purpose of this study is to investigate the parental marital relationships and the parent-child relationships of child psychiatric parents and its control group. This study was carried out two questionnaire instruments ; The marital satisfaction inventory(MSI) and the dyadic adjustment scale(DAS). The subjects are parents of the child psychiatric patients. A matched control group and parents of child psychiatric parents in Seoul area which were collected from July 1987 to September 1987, and classified into five subgroups : 1 Psychiatric disorder 2) Neurotic disorder 3) Tic disorder 4) Autistic disorder 5) Mental retardation. The results are as following ; 1) M.S.I scale scores of parents of patients group are lower than that of control group. 2) D.A.S scale score of parents of patients group is significantly lower than that of the control group(P<0.01). 3) The global distress scale(GDS) of the M.S.I. was most positively correlated with affective communication(AFC) and problem-sloving communication(PSC). 4) Female shoed more modern concept of role identification than male but tend to have heavier role assignment especially in child rearing practices which could be characterized by maternal domination. 5) Affective communication and sexual relationship between married couple and child rearing practices are influenced by their own family history of distress. 6) The marital global distress scale(GDS) score was highest in the parents of psychosis, the next in the parents of neurosis, autism, mental retardation, and tic disorder in descending order of severity. 7) The dyadic maladjustment score was highest in the parents of psychosis, the next in the parents of neurosis, tic, autism and mental retardation in descending order of severity. 8) Conflict in child rearing and parenting problems were particularly prominent in parents of the tic patients, and their marital relationship was not significantly disturbed. The above finding suggested that couple adjustment and marital dissatisfaction were closely related with child rearing problems and the children's disorder. So marital dissatisfaction and marital maladjustment seem to play a significant role in the genesis of psychosis and neurosis not much in autism and mental retardation.

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