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SCIATIC NERVE REGENERATION USING CALCIUM PHOSPHATE COATED CONDUIT AND BRAIN-DERIVED NEUROTROPHIC FACTOR GENE-TRANSFECTED SCHWANN CELL IN RAT (인회석 박막 피복 도관과 Brain-derived neurotrophic factor(BDNF) 유전자 이입 슈반세포를 이용한 백서 좌골신경 재생에 관한 연구)

  • Choi, Won-Jae;Ahn, Kang-Min;Hwang, Soon-Jeong;Choung, Pill-Hoon;Kim, Myung-Jin;Kim, Nam-Yeol;Yoo, Sang-Bae;Jahng, Jeong-Won;Kim, Hyun-Man;Kim, Joong-Soo;Kim, Yun-Hee;Kim, Soung-Min;Lee, Jong-Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.31 no.3
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    • pp.199-218
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    • 2005
  • Purpose of Study: Peripheral nerve regeneration depends on neurotrophism of distal nerve stump, recovery potential of neuron, supporting cell like Schwann cell and neurotrophic factors such as BDNF. Peripheral nerve regeneration can be enhanced by the conduit which connects the both sides of transected nerve. The conduit maintains the effects of neurotrophism and BDNF produced by Schwann cells which can be made by gene therapy. In this study, we tried to enhance the peripheral nerve regeneration by using calcium phosphate coated porous conduit and BDNF-Adenovirus infected Schwann cells in sciatic nerve of rats. Materials and Methods: Microporous filter which permits the tissue fluid essential for nerve regeneration and does not permit infiltration of fibroblasts, was made into 2mm diameter and 17mm length conduit. Then it was coated with calcium phosphate to improve the Schwann cell adhesion and survival. The coated filter was evaluated by SEM examination and MTT assay. For effective allogenic Schwann cell culture, dorsal root ganglia of 1-day old rat were extracted and treated with enzyme and antimitotic Ara-C. Human BDNF cDNA was obtained from cDNA library and amplified using PCR. BDNF gene was inserted into adenovirus shuttle vector pAACCMVpARS in which E1 was deleted. We infected the BDNF-Ad into 293 human mammary kidney cell-line and obtained the virus plaque 2 days later. RT-PCR was performed to evaluate the secretion of BDNF in infected Schwann cells. To determine the most optimal m.o.i of BDNF-Ad, we infected the Schwann cells with LacZ adenovirus in 1, 20, 50, 75, 100, 250 m.o.i for 2 hours and stained with ${\beta}$-galactosidase. Rats(n=24) weighing around 300g were used. Total 14mm sciatic nerve defect was made and connected with calcium phosphate coated conduits. Schwann cells$(1{\times}10^6)$ or BDNF-Ad infected Schwann cells$(1{\times}10^6)$ were injected in conduit and only media(MEM) was injected in control group. Twelve weeks after surgery, degree of nerve regeneration was evaluated with gait analysis, electrophysiologic measurements and histomorphometric analysis. Results: 1. Microporous Millipore filter was effective conduit which permitted the adhesion of Schwann cells and inhibited the adhesion of fibroblast. We could enhance the Schwann cell adhesion and survival by coating Millipore filter with calcium phosphate. 2. Schwann cell culture technique using repeated treatment of Ara-C and GDNF was established. The mean number of Schwann cells obtained 1 and 2 weeks after the culture were $1.54{\pm}4.0{\times}10^6$ and $9.66{\pm}9.6{\times}10^6$. 3. The mRNA of BDNF in BDNF-Ad infected Schwann cells was detected using RT-PCR. In Schwann cell $0.69\;{\mu}g/{\mu}l$ of DNA was detected and in BDNF-Adenovirus transfected Schwann cell $0.795\;{\mu}g/{\mu}l$ of DNA was detected. The most effective infection concentration was determined by LacZ Adenovirus and 75 m.o.i was found the most optimal. Conclusion: BDNF-Ad transfected Schwann cells successfully regenerated the 14mm nerve gap which was connected with calcium phosphate coated Millipore filter. The BDNF-Ad group showed better results compared with Schwann cells only group and control group in aspect to sciatic function index, electrophysiologic measurements and histomorphometric analysis.

The Effects of Prostaglandin and Dibutyryl cAMP on Osteoblastic Cell Activity and Osteoclast Generation (Prostaglandin과 Dibutyryl cAMP가 조골세포의 활성과 파골세포 형성에 미치는 영향)

  • Mok, Sung-Kyu;You, Hyung-Keun;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.26 no.2
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    • pp.448-468
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    • 1996
  • To maintain its functional integrity, bone is continuously remodelled by a process involving resorption by osteoeclasts and formation by osteoblasts, In order to respond to changes in the physical environment or to trauma with the relevant action, this process is strictly regulated by locally synthesized or systemic fators, Prostaglandin $E_2(PGE_2$) is perhaps one of the best studied factors, having been known to affect bone cell function for several decades.$PGE_2$ has both anabolic and catabolic activities. Excess of $PGE_2$ has been implicated in a number of pathological states associated with bone loss in a number of chronic inflammatory conditions such as periodontal disease and rheumatoid arthritis. $PGE_2$ and other arachidonic acid metabolites have been shown to be potent stimulators of osteoclastic bone resorption in organ culture. The anabolic effects of $PGE_2$ were first noticed when an increase in periosteal woven bone formation was seen after the infusion of $PGE_2$ into infants in order to prevent closure of the ductus arteriosus. The cellular basis for the catabolic actions of $PGE_2$ has been well characterized. $PGE_2$increases osteoclast recruitment in bone marrow cell cultures. Also $PGE_2$ has a direct action on osteoclast serving to inhibit activity and can also indirectly activate osteoclast via other cells in the vicinity, presumably osteoblast. The cellular mechanisms for the anabolic actions of $PGE_2$ are not nearly so well understood. The purpose of this paper was to study the effects of $PGE_2$ and dibutyl(DB)cAMP on osteoblastic clone MC3T3El cells and on the generation of osteoclasts from their precursor cells. The effect of $PGE_2$ and DBcAMP on the induction of alkaline phoaphatase(AlP) was investigated in osteoblastic clone MC3T3El cells cultured in medium containing 0.4% fetal bovine serum. $PGE_2$ and DBcAMP stimulated ALP activity and MTT assay in the cells in a dose-dependent manner at concentrations of lO-SOOng/ml. Cycloheximide, protein synthesis inhibitor, inhibited the stimulative effect of $PGE_2$ and DBcAMP on ALP activity in the cells. $PGE_2$also increased the intracellular cAMP content in a dose-dependent fashion with a maximal effect at 500ng/ml. The effect of $PGE_2$ on the generation of osteoclasts was investigated in a coculture system of mouse bone marrow cells with primary osteoblastic cells cultured in media containing 10% fetal bovine serum.After cultures, staining for tartrate-resistant acid phosphatase(TRAP)-marker enzyme of osteoclast was performed. The TRAP(+) multinucleated cells(MNCs), which have 3 or more nuclei, were counted. More TRAP(+) MNCs were formed in coculture system than in control group. $PGE_2(10^{-5}10^{-6}M)$ stimulated the formation of osteoclast cells from mouse bone marrow cells in culture. $PGE_2(10^{-6}M)$ stimulated the formation of osteoclast cells from mouse bone marrow cells in coculture of osteoblastic clone MC3T3E1 cells This results suggest that $PGE_2$ stimulates the differentiation of osteoblasts and generation of osteoclast, and are involved in bone formation, as well as in bone resorption.

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Chemical Compositions of Fermented Codonopsis lanceolata (발효더덕의 화학성분)

  • Park, Sung-Jin;Seong, Dong-Ho;Park, Dong-Sik;Kim, Seung-Seop;Gou, Jing-Yu;Ahn, Ju-Hee;Yoon, Won-Byung;Lee, Hyeon-Yong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.3
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    • pp.396-400
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    • 2009
  • Dodok (Codonopsis lanceolata Bench. et Hook) root contains abundant pharmaceutical substances and is widely used as a food and a medicinal herb. To identify the major components, fermented Codonopsis lanceolata was analyzed for its chemical compositions prior to their pharmaceutical substances, which were used as the fundamental data. The contents of carbohydrate, crude protein, crude lipid and ash are 79.3%, 13.0%, 2.40% and 5.3%, respectively. The calories of fermented Codonopsis lanceolata was 390.5 kcal. Total dietary fiber was 47.4% of total carbohydrates. The protein was composed of 18 different amino acids. The contents of essential and non-essential amino acids were 8,118.18 mg and 10,913.42 mg. The K was the largest mineral followed by P, Ca and Mg, which means fermented Codonopsis lanceolata is alkali material. The contents of saturated fatty acids, monounsaturated fatty acids and polyunsaturated fatty acids were 37.76%, 3.84%, and 35.64%, respectively. Therefore, the amount of the total unsaturated fatty acid was higher than that of any other plant. The content of crude saponin in fermented Codonopsis lanceolata was 60.1 mg/g. It is expected that a follow up study on fermented Codonopsis lanceolata through development and evaluation of processed foods for their functional properties would provide useful information as a source of medicinal foods.

Biological Activities of Extracts from Gamma-irradiated Aralia elata Cortex (감마선 조사한 총목피(Aralia elata Cortex) 추출물의 생리활성)

  • Park, Hye-Jin;Lee, Eun-Ho;Kim, Myung-Uk;Lee, Seon-Ho;An, Dong-Hyun;An, Bong-Jeun;Kwon, Joong-Ho;Cho, Young-Je
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.8
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    • pp.1236-1247
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    • 2014
  • Gamma irradiated-treatment of natural medicinal plants can be used to improve extraction transference number and for qualitative improvement of color when applied to functional material exploration. This study investigated the biological activities of Aralia elata cortex extracts upon gamma irradiation. In addition, different physical techniques [photostimulated luminescence (PSL) and thermoluminescence (TL)] were used for irradiation identification of Aralia elata cortex. In PSL analysis, non-irradiated (0 kGy) sample showed a negative result of 400 photon counts (PCs), whereas irradiated (5, 10, and 30 kGy) samples showed positive results of 90,100.00, 312,614.33, and 321,661.67 PCs, respectively. In the TL method, growth curve showed very unusual behaviors around $200^{\circ}C$ upon natural-irradiation of the non-irradiated (0 kGy) sample and around $150{\sim}250^{\circ}C$ for the irradiated (5, 10, and 30 kGy) samples. The TL ratio was 0.1 in non-irradiated samples at 0.011, whereas the values of irradiated samples (5, 10, and 30 kGy) were 0.1 at 1.105, 1.009, and 2.206, respectively. For phenolics of gamma-irradiated Aralia elata cortex, water and 50% ethanol extracts had the highest amounts, $17.30{\pm}0.40mg/g$ and $18.87{\pm}0.46mg/g$ at 10 kGy irradiation, respectively. The inhibitory activities of angiotensin-converting enzyme and xanthin oxidase were higher in both irradiated water and 50% ethanol extracts than in non-irradiated ones. For pancreatin ${\alpha}$-amylase and ${\alpha}$-glucosidase inhibitory activities, water and 50% ethanol extracts containing $200{\mu}g/mL$ of phenolics showed high inhibitory activities of 60~100% at all irradiation doses (0~30 kGy). This result confirmed that Aralia elata cortex extracts have greater anti-diabetic effects than acabose as a diabetic remedy. Gamma-irradiated Aralia elata cortex extracts are useful as a functional material with anti-diabetic effects. Thus, Aralia elata cortex extracts can be used as a functional material with various biological activities, and gamma-irradiation can be used to amplify biological activities in plants.

Gastrointestinal Stromal Tumor (GIST) of the Stomach: Clinicopathologic Analysis and Outcome (위에 발생한 위장관 간질성 종양의 임상병리학적 특성과 치료성적)

  • Ryu Je-Seock;Lee Sung-Ryul;Choi Sae-Byeol;Park Sung-Soo;Lee Ju-Han;Kim Seung-Joo;Kim Chong-Suk;Chae Yang-Seok;Mok Young-Jae
    • Journal of Gastric Cancer
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    • v.5 no.1
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    • pp.40-46
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    • 2005
  • Purpose: Gastrointestinal stromal tumors (GISTs) are mesenchymal neoplasms of the gastrointestinal tract. GISTs are positive for the expression of c-Kit protein at immunohistochemistry, and their clinical presentations vary. This retrospective study was performed to evaluate the clincopathologic characteristics of GISTs and to define the prognostic factors. Materials and Methods: 40 patients who underwent a complete resection of a GIST during the period $1996\~2003$ at the Department of Surgery, Korea University College of Medicine, were studied. We divided them into low- and high-risk. groups by using tumor size and mitotic count: 23 cases were low risk, and 17 were high risk. Clinicopathologic features, immunohistochemical findings, and prognoses were compared between the low- and the high-risk groups. Results: The mean age of the 40 patients was $61.3\pm11.1$years, and the male-to-female ratio was 1:1.1. There was no significant difference in age and sex between the groups. A comparative analysis revealed tumor size, mitotic count, clinical symptoms, preoperative pathologic diagnosis, ulceration, and necrosis to be variables that had statistically significant differences between the high- and the low-risk groups. In the univariate analysis, tumor size, mitotic count, ulceration, necrosis, and abnormal endoscopic ultrasound findings were associated with disease-free survival, but in the multivariate analysis, mitotic activity was the only independent factor associated with disease-free survival. 8 patients had recurrences during the follow-up period, and four of them were treated with STI-571 (imatinib mesylate, $Gleevec^{(R)}$). The treated patients have survived until now; however, two of non-treated patients died from disease progression. Conclusion: Based on this study, tumor size, ulceration, and necrosis are significant factors affecting survival, and mitotic activity may be a useful prognostic marker. STI-571 may be used in an adjuvant setting because the drug has shown anticancer activity in patients with recurrence or metastasis.

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ULTRASTRUCTURAL ANALYSIS OF TOOTH PULP AFFERENTS TERMINALS IN THE MEDULLARY DORSAL HORN OF THE RAT (치수유래 구심성 신경섬유의 삼차신경 감각핵군에서의 연접특성)

  • Bae, Yong-Chul;Lee, Eun-Hee;Choy, Min-Ki;Hong, Su-Hyung;Kim, Hyun-Jung;Na, Soon-Hyeun;Kim, Young-Jin
    • Journal of the korean academy of Pediatric Dentistry
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    • v.28 no.2
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    • pp.219-227
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    • 2001
  • Little is known about processing mechanism of pain sensation of the oral cavity at the 1st synapse of trigeminal sensory nuclei. Serial ultrathin sections of tooth pulp afferent terminals, identified by the transganglionic transport of 1% wheatgerm agglutinin conjugated horseradish peroxidase, were investigated with electron microscope. Quantitative ultrastructural analysis was performed on digitizing tablet connected to Macintoshi personal computer (software; NIH Image 1.60, NIH, Bethesda, MD). Labeled boutons could be classified into two types by the shapes of containing vesicles : S bouton, which contained mainly spherical vesicles (Dia. 45-55 nm) and few large dense cored vesicles (Dia, 80-120nm), and LDCV bouton, which contained spherical vesicles as well as large number of large dense cored vesicles. Most of the parameters on the ultrastructural characteristic and synaptic organization of labeled boutons were similar between S and LDCV boutons, except shapes of containing vesicles. Majority of the labeled boutons showed simple synaptic arrangement. The labeled boutons were frequency presynaptic to dendritic spine, and to a lesser extent, dendritic shaft. They rarely synapsed with soma and adjacent proximal dendrite. A small proportion of labeled boutons made synaptic contacts with presynaptic, pleomorphic vesicles containing endings and synaptic triad. Morphometric parameters of labeled boutons including volume and surface area, total apposed area, mitochondrial volume, active zone area, vesicle number and density showed wide variation and these were not significantly different between S and LDCV boutons. The present study revealed characteristic features on ultrastructure and synaptic connection of pulpal afferents which may involved in transmission of oral pain sensation.

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Intestinal Atresia - The Second National Survey (선천성 장폐쇄증 2차 조사 - 대한소아외과학회 회원을 주 대상으로 한 전국조사 -)

  • Kim, Seong-Chul;Kim, D.Y.;Kim, S.Y.;Kim, I.K.;Kim, I.S.;Kim, J.E.;Kim, J.C.;Kim, H.Y.;Park, K.W.;Park, W.H.;Park, J.Y.;Seo, J.M.;Seol, J.Y.;Oh, S.M.;Oh, J.T.;Lee, N.H.;Lee, M.D.;Lee, S.K.;Lee, S.C.;Chung, S.Y.;Jung, S.E.;Jeong, Y.J.;Jung, P.M.;Choi, K.J.
    • Advances in pediatric surgery
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    • v.16 no.1
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    • pp.1-10
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    • 2010
  • The members of the Korean Association of Pediatric Surgeons conducted a retrospective study of two hundred and twenty-two cases of intestinal atresia for the period from January 1, 2007 to December 31, 2009. Seventeen hospitals were involved. There were 76 duodenal, 65 jejunal, and 81 ileal atresias (3 colonic). The male to female ratio was 0.85:1 in DA and 1.34:1 in JIA. Ninety-four patients(43.3 %) were premature babies (DA 40.3 %, JA 64.6 %, IA 28.8 %), and 70 babies (32.0 %) had low birth weight (DA 38.7 %, JA 44.4 %, IA 16.0 %). Antenatal diagnosis was made in 153 cases (68.9 %). However, 27 infants (17.6 %) with antenatal diagnosis were transferred to the pediatric surgeon's hospitals after delivery. Maternal polyhydramnios was observed in 81 cases (36.59 %) and most frequent with proximal obstruction. In forty-four cases (19.8 %), only simple abdominal film was taken for diagnostic study. The associated malformations were more frequently observed in DA - 61.8 % in DA and 22.6 % in JIA. Meconium peritonitis, small bowel volvulus and intussusception were more frequently associated with ileal atresia. The overall mortality rate was 3.6 %. (Abbreviations: DA;duodenal atersia, JA;jejunal atresia, JIA;jejunoileal atresia, IA;ileal atrsia).

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Acute Appendicitis -A Survey by the Korean Association of Pediatric Surgeons in 2006- (급성 충수염 - 2006년 대한소아외과학회회원 대상 전국조사 -)

  • Lee, Suk-Koo;Kim, D.Y.;Kim, S.Y.;Kim, S.C.;Kim, S.G.;Kim, W.K.;Kim, I.K.;Kim, J.E.;Kim, J.C.;Park, K.W.;Park, W.H.;Seo, J.M.;Song, Y.T.;Oh, J.T.;Lee, N.H.;Lee, D.S.;Lee, M.D.;Lee, S.C.;Chang, S.I.;Jun, Y.S.;Chung, S.Y.;Chung, S.E.;Chung, U.S.;Jung, P.M.
    • Advances in pediatric surgery
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    • v.13 no.2
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    • pp.203-211
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    • 2007
  • A nationwide survey on acute appendicitis (AAP) was undertaken among members of the Korean Association of Pediatric Surgeons. The members were required to complete a questionnaire and the case registration form for each patient during the 6 months period from the $1^{st}$ October, 2005 to $31^{st}$ March, 2006. Questionnaires were collected from 23 members. Four hundred seventy six patients were registered from 21 hospitals where the members were working. The male to female ratio was 1.67:1. Average age was 9 years and 5 months (range 21 months-20 years). Operations were performed on average 10.4 hours (range 1-230 hours) from arrival at hospital. The average operation time was 59.1 minutes. The average admission days were 5.8 days (range 2-45 days). The most frequent symptoms were abdominal pain (95.1 %), vomiting (50.6 %) and fever (43.7%). The average duration of symptoms was 42.2 hours (range 1 hour-22 days). Leukocytosis (WBC count > 10,000) was found in 85.9%. The most popular diagnostic tools were ultrasonography and CT. Open surgery was performed in 72.1 % and laparoscopic surgery in 27.5 %. Two laparoscopic surgeries were converted to open surgery (0.4 %). Simple appendicitis was found in 54.5 % and complicated appendicitis such as abscess, gangrenous change and perforation in 45.5%. According to the questionnaires 12 hospitals were performing open surgery only in all patients. Four hospitals were performing laparoscopic surgery in all patients. Seven hospitals are performing both surgical methods according to surgeon or occasion. Regarding the use of antibiotics in acute appendicitis, three kinds of antibiotics were used in 40 % of total simple appendicitis patients. The results showed the trend of management in acute appendicitis in Korea. Especially it is necessary for the members to discuss the use of antibiotics for prophylaxis in the simple appendicitis.

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Nutrition Intake according to Food and Exercise Habits in Female College Students of Yang-Ju si (경기도 양주시 여대생의 식습관, 운동습관, 건강관련행동이 영양섭취수준에 미치는 영향)

  • Rha, Young Ah;Kang, Min Jeong;Lee, Sun Ho;Kim, Jung Yun
    • Culinary science and hospitality research
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    • v.21 no.4
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    • pp.284-293
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    • 2015
  • This study was intended to investigate food, exercise habits, perception of health status and the effects of nutrient intake of female college students. Food habits such as regularity of meal time, repast of breakfast, numbers of night snack per week and duration of meal were assessed via a self reporting questionnaire, and 24hrs recall method for nutrient intake was obtained from 155 female college students in Yangju-si. The mean height and weight of the subjects were $161.25{\pm}3.52cm$ and $52.26{\pm}4.52kg$, respectively. The average intakes of energy and nutrients were below the Korea recommended dietary allowance(RDA). Especially, The nutrient intakes(energy, protein, fat, fiber, calcium, iron, phosphorus, zinc, sodium, potassium, vitamin B6, niacin, vitamin C, folate, vitamin E, cholesterol) of the group in which the regularity of meal time per week was 5~7 days was significantly higher than the group in which the regularity of meal time was < 3 days (p<0.005). The nutrient intakes influenced by the food habit such as the regularity of meal time, skipping breakfast, and the numbers of night snack per week but did not differ in accordance with the regular exercise habits and perception of health status. These findings suggest that nutritional education based on female college students' food habits(regularity of meal time, skipping breakfast and night snacks per week, and duration of meal) may be required to improve dietary variety. It is considered that the improvement in dietary habits will contribute to the improvement of nutrition.

The Relation between Mother's LOHAS Propensity and Healthy Dietary Life Attitude perceived by Chinese University Students (대학생 자녀가 지각한 어머니의 로하스(LOHAS) 성향과 건강식생활 태도와의 관련성 연구 - 중국대학생을 중심으로 -)

  • Lee, Yeon-Jung;Ahn, Gee-Jung
    • Culinary science and hospitality research
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    • v.21 no.4
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    • pp.101-120
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    • 2015
  • This study was conducted to examine the relation between a mother's LOHAS lifestyle propensity, healthy dietary life, and nutrient intake perceived by Chinese university students. The subjects consisted of 376 Chinese university students. The research results are summarized below. The mother's LOHAS attitude index was 76.02 points out of a perfect score of 100 points. High LOHAS attitude factors of mothers were 'health', whereas 'social welfare' scored low. The highest LOHAS attitude items of mothers that Chinese university students perceived was 'My mother uses health food, organic food, clean food'(4.26 points). Males who lived on their own in a large city and have normal BMI ($18.5{\sim}22.9kg/m^2$) perceived a high mother's LOHAS score. The highest food habit variables are 'eating without spilling food' 'cleaning up after meals around' and 'washing my hands before meals'. The highest healthy dietary life item is 'I try to maintain a pleasant and enjoyable life'(3.83 points). The highest nutrient intake attitude score is 'I tend to eat yellowish-green vegetables every day.'(3.92 points). According to mother's LOHAS lifestyle score, it was verified that the higher mother's LOHAS lifestyle score, the higher children's food habit, healthy dietary life, and nutrient intake score are. Mother's environment-friendly LOHAS attitude index has a meaningful impact on the child's food habits, healthy dietary life, and nutrient intake.