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Purification and Enzymatic Characteristics of the Bacillus pasteurii Urease Expressed in Escherichia coli (Escherichia coli에서 발현된 Recombinant Bacillus pasteurii Urease의 정제 및 효소학적 특성)

  • 이은탁;김상달
    • Microbiology and Biotechnology Letters
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    • v.20 no.5
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    • pp.519-526
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    • 1992
  • The gene coding for urease of alkalophilic Bacillus pasteurii had been cloned in Escherichia coli previously. The urease protein was purified 63.1-fold by TEAE-cellulose, DEAE-Sephadex A-50, Sephadex G-150 and Sephadex G-200 chromatographies with a 7.3% yield from the sonicated fluid of the E. coli HB1Ol(pBUll) encoding B. pasteurii urease gene. The ureases of E. coli (pBUll) and B. pasteurii possessed as a $K_m$ for urea, 42.1 mM and 40.4 mM, respectively. They hydrolyzed urea with $V_{max}$ of 86.9$\mu$mol/min and 160$\mu$mol/min, respectively. Both ureases were composed with four subunits (Mrs 67,000) and a subunit (Mr 20,000). The molecular weight of both native enzymes was Mr 280,OOO$pm$10,000 determined by gel filtration chromatography and Coomassie blue staining of the subunits. The optimal reaction pH of both ureases were pH 7.5. The ureases were stabled in pH 5.5-10.5. The optimal reaction temperature of both ureases were $60^{\circ}C$, and the ureases were stable for an hour at $50^{\circ}C$, 40min at $60^{\circ}C$ and 10 min at $70^{\circ}C$ The activity of both enzymes were inhibited completely by $Ag^{2+}$, $Hg^{2+}$, $Zn^{2+}$, $Cu^{2+}$, and were inhibited 60% by CoH, 30% by $Fe^{2+}$ and 10% by $Pb^{2+}$. However it was increased by the addition of $Sn^{2+}$, $Mn^{2+}$, $Mg^{2+}$ at concentration of $1{\times}10^{-3}$M. Both ureases were inhibited completely by p-CMB and acetohydroxamic acid. The urease expressed in E. coli (pBU11) was inhibited 70% by SDS. The urease of B. pasteurii was inhibited 40% by hydroxyurea, whereas the recombinant urease of E. coli strain was inhibited 17%. Both enzymes were not inhibited by cyclohexanediaminetetraacetic acid (CDTA) and ethylendiaminetetraacetic acid (EDTA).

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A Study Related to Adolescent Students' School Uniform Behavior and Attitude toward Appearance (청소년의 교복행동과 외모에 대한 태도와의 관련 연구)

  • Han, Mi-Hwa;Lee, Eun-Hee
    • Journal of Korean Home Economics Education Association
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    • v.21 no.2
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    • pp.23-43
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    • 2009
  • This study examined the relativity between school uniform behavior(especially attitude toward school uniform, school uniform satisfaction, school uniform alteration) and attitude toward appearance in the adolescence. From November to December in 2007, 898 middle school and high school students from four schools in Jeollabuk-do Province were analyzed and the influence of relevant variables was understood to gather basic information about the fashion life in the adolescence. This study used SPSS 11.5 for Windows Program to conduct frequency analysis, factor analysis, reliability analysis, ${\chi}^2$-test, t-test, One-way ANOVA, Duncan's multiple comparison test, and Pearson's correlation. The following summarizes the findings of this study: The results of analysing the factors to the response attitude toward uniforms and attitude toward appearance and clothing attitude emerged four dimensions(fashion, symbolism, cleanliness, alternation), three dimensions(needs value conformity toward appearance). In attitude toward uniform, female students were more actively altering their uniforms than male students and middle school students were more sensitive to trends than high school students. However, high school students were more aware of the cleanliness and alteration of uniforms. Overall, most students were not very satisfied with their uniforms (design, color, texture). When students' attitude toward appearance and demographical characteristics were examined, it was found that female students were more aware of desire, value, and conformity in appearances than male students. By age, it was found that high school students were more aware than middle school students. Also, students receiving KRW 30,000 or more for monthly allowance were more aware than others who receive a lower amount between KRW 10,000 and KRW 20,000. Therefore, most students(62.1%) have experiences in altering uniforms. Especially, more female students and more high school students had such experiences. Uniform alteration is more related to attitude to appearance. In other words, students who choose to alter their uniforms are highly aware of desire, value, and conformity toward appearance. Students who are satisfied with all factors regarding attitude toward uniform, except for alteration, did not alter their uniforms that much. In addition, when the relativity between students' attitude toward appearance with uniform was considered, students that are not very satisfied with their uniforms were more aware of desire, value, and conformity toward appearance. In conclusion, it was found that students' attitude toward appearance and school uniform behavior are closely related and their interests in appearance lead to alteration of uniforms, the clothing in which they spend most time of the day. From now on, students' opinions shall be considered when selecting or designing school uniforms.

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Relationships between Fatty Acids and Tocopherols of Conventional and Genetically Modified Peanut Cultivars Grown in the United States (미국산 전통품종과 유전자 재조합 땅콩품종의 지방산과 토코페롤의 상관관계)

  • Shin, Eui-Cheol
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.10
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    • pp.1618-1628
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    • 2013
  • Relationships between fatty acids and tocopherols in conventional and genetically modified peanut cultivars were studied by gas chromatography with flame ion detector and high performance liquid chromatography with fluorescence detection. Eight fatty acids and four tocopherol isomers in the sample set were identified and quantified. Oleic acid and linoleic acid are major fatty acids and the ratio of oleic and linoleic acids ranged from 1.11 to 16.26. Tocopherols contents were 6.76 to 12.24 for ${\alpha}$-tocopherol (T), 0.08 to 0.39 for ${\beta}$-T, 5.28 to 15.02 for ${\gamma}$-T, and 0.17 to 1.17 mg/100 g for ${\delta}$-T. Correlation coefficient (r) for fatty acids and tocopherols indicated a strong inverse relationship between oleic & linoleic acids (r=-0.97, P<0.05) and positive relationships between palmitic & linoleic acids (r=0.95, P<0.05) and ${\gamma}$-T & ${\delta}$-T (r=0.83, P<0.05). Principal component analysis (PCA) of fatty acids and tocopherols gave four significant principal components (PCs, with eigenvalues>1), which together account for 85.49% of the total variance in the data set with PC1 and PC2 contributing 45.27% and 21.33% of the total variability, respectively. Eigen analysis of the correlation matrix loadings of the four significant PCs revealed that PC1 was mainly contributed by palmitic, oleic, linoleic, and gondoic acids, while PC2 was by behenic acid, ${\beta}$-T, and ${\gamma}$-T. The score plot generated by PC1-PC2 identified sample clusters in the two spatial planes based on the oleic and linoleic acids. The score plot PC3-PC4 didn't separate sample groups.

A Study on the Care Needs of Family-Caregivers and the Level of Self Care for Patients of Cerebral Vascular Accident(CVA) (뇌졸중환자의 자가간호 수준과 가족구성원의 간호요구)

  • Cho Young-Hee
    • Journal of Korean Academy of Fundamentals of Nursing
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    • v.7 no.2
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    • pp.239-255
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    • 2000
  • The purpose of this study was to explore the care needs of family-caregivers caring for patients with a CVA and the level of self care of the patients. The subjects for the study were 112 patients with a CVA and their caregivers. These patients were seen in a hospital or out-patient-department(OPD) at two oriental medical hospital in Jeonbuk province. The survey instruments used in this study were Kang's ADL checklist for self care of patients and Kim's Likert-style checklist for care needs of family-caregivers to patient with CVA. The survey was conducted from July 4 to August 30, 1999. Internal validity by calculation of Cronbach's alpha was 0.95, which was regarded as high. The survey results were analyzed using the SPSS program, with percentages, means, t-test, ANOVA and Pearson's correlation coefficients. The results of this study are as follows : 1. The level of self care for patients with a CVA was : 1) complete dependence(M=14.9, 13.1%), 2) complete independence(M=23.6, 20.9%), 3) incomplete independence(M=23.9, 21.0%), 4) incomplete dependence(M=26.6, 25.0%), 5) dependence and independence(M=23.0, 20.0%). The items for which there was a high level of self care were : 1) drinking(M=3.62), 2) eating (M=3.25), 3) position returning(M=3.18) : and the items for which there was a low level of self care were : 1) ascending and descending stairs(M=2.08), 2) walking(M=2.47). 3) putting on and taking off trousers(M=2.55). 2. The mean score of the sum of the care needs of the family-caregivers was : 1) need for immediate care and help: 2) need of the way to communicate with patient: 3) need for education and assistance related to physical functional level: 4) need to be informed about the disease, treatment and care: 5) need for social support and consultation: 6) need for appreciation: 7) need for management of nursing problems related to immobility. The highest meed factor was the need for immediate care and help(M=3.47): and lowest need factor was the need for management of nursing problems related to immobility(M=2.80). 3. There were significant differences between the level of care need and general characteristic of the caregivers, there were family-caregivers age(P=0.001), marital status (P=0.276), occupation (P=0.006), monthly income(P=0.000), Patient's relationship to caregivers(P=0.004) and health(P=0.000). 4. There were significant differences between the level of self care and general characteristic of the patients, there were patient paralytic condition(P=0.01), blood pressure(P=0.01), and length of suffering(P=0.03). 5 There were significant differences between the level of care need and the general characteristic factors, which were CVA patient's blood pressure (P=0.05), problem of medical fee (P=0.05). 6. There was significantly correlation with the family-caregivers care need and the level of self care in the CVA patient(r=0.300, P=0.000). As a result, need to promote the level of self care in patients and to meet the care need of family-caregivers for more efficient nursing of CVA patients, is emphasized. Therefore more study is needed on an efficient way to provide rehabilitation and quality nursing interventions for family-caregivers and patients with CVA.

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Composition-Some Properties Relationships of Non-Alkali Multi-component La2O3-Al2O3-SiO2 Glasses (무알칼리 다성분 La2O3-Al2O3-SiO2 유리의 조성과 몇 가지 물성의 관계)

  • Kang, Eun-Tae;Yang, Tae-Young;Hwang, Jong-Hee
    • Journal of the Korean Ceramic Society
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    • v.48 no.2
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    • pp.127-133
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    • 2011
  • Non-Alkali multicomponent $La_2O_3-Al_2O_3-SiO_2$ glasses has been designed and analyzed on the basis of a mixture design experiment with constraints. Fitted models for thermal expansion coefficient, glass transition temperature, Young's modulus, Shear modulus and density are as follows: ${\alpha}(/^{\circ}C)=8.41{\times}10^{-8}x_1+5.72{\times}10^{-7}x_2+2.13{\times}10^{-7}x_3+1.09{\times}10^{-7}x_4+1.10{\times}10^{-7}x_5+1.15{\times}10^{-7}x_6+2.72{\times}10^{-8}x_7+2.41{\times}10^{-7}x_8-1.08{\times}10^{-8}x_1x_2+4.28{\times}10^{-8}x_3x_7-2.02{\times}10^{-8}x_3x_8-1.60{\times}10^{-8}x_4x_5-2.71{\times}10^{-9}x_4x_8-2.19{\times}10^{-8}x_5x_6-3.89{\times}10^{-8}x_5x_7$ $T_g(^{\circ}C)=7.36x_1+15.35x_2+20.14x_3+8.97x_4+13.85x_5+4.22x_6+28.21x_7-1.44x_8-0.84x_2x_3-0.45x_2x_5-1.64x_2x_7+0.93x_3x_8-1.04x_5x_8-0.48x_6x_8$ $E(GPa)=2.04x_1+14.26x_2-1.22x_3-0.80x_4-2.26x_5-1.67x_6-1.27x_7+3.63x_8-0.24x_1x_2-0.07x_2x_8+0.14x_3x_6-0.68x_3x_8+0.29x_4x_5+1.28x_5x_8$ $G(GPa)=0.35x_1+1.78x_2+1.35x_3+1.87x_4+9.72x_5+29.16x_6-0.99x_7+3.60x_8-0.48x_1x_6-0.50x_2x_5+0.08x_3x_7-0.66x_3x_8+0.94x_5x_8$ ${\rho}(g/cm^3)=0.09x_1+0.51x_2-4.94{\times}10^{-3}x_3-0.03x_4+0.45x_5-0.07x_6-0.10x_7+0.07x_8-9.60{\times}10^{-3}x_1x_2-8.20{\times}10^{-3}x_1x_5+2.17{\times}10^{-3}x_3x_7-0.03x_3x_8+0.05x_5x_8$ The optimal glass composition similar to the thermal expansion coefficient of Si based on these fitted models is $65.53SiO_2{\cdot}25.00Al_2O_3{\cdot}5.00La_2O_3{\cdot}2.07ZrO_2{\cdot}0.70MgO{\cdot}1.70SrO$.

Development of Optimal Culture Media for the Stable Production of Mushroom (버섯의 안정적 생산을 위한 최적배지의 개발)

  • Gal, Sang-Wan;Lee, Sang-Won
    • Applied Biological Chemistry
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    • v.45 no.2
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    • pp.71-76
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    • 2002
  • Several antagonistic bacteria, SD-1, 4, 10, 11, 14, 15, and 16, which have strong CMCase and amylase activities, were isolated from the fermented mushroom media. Among them, SD-1, 10, 11, and 15 have strong antibacterial activities against the mushroom pathogenic bacteria Pseudomonas sp., and SD-1, 10, 11, 14, and 16 have strong antifungal activities against the mushroom pathogenic fungi, Trichoderma sp. SD-14, 15, and 16 did not inhibit the growth of mushroom Pleurotus eryngii ASI-2302, and Pleurotus ostreatus ASI-2042 and ASI-2180. When the culture broth mixture of the seven bacterial strains was applied to the mushroom media, the growths of pathogens, Pseudomonas sp. and Trichoderma sp., were inhibited.

Enhancement of Immunomodulatory Activities of Low Molecular Weight Fucoidan Isolated from Hizikia fusiforme (톳 유래 저분자 푸코이단의 면역활성 증진)

  • Ha, Ji-Hye;Kwon, Min-Chul;Han, Jae-Gun;Jin, Ling;Jeong, Hyang Suk;Choi, Geun-Pyo;Park, Uk-Yeon;You, Sang-Guan;Lee, Hyeon-Yong
    • Korean Journal of Food Science and Technology
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    • v.40 no.5
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    • pp.545-550
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    • 2008
  • The anticancer and immunomodulatory activities of low molecular weight (Mw 11 kDa) fucoidan isolated from Hizikia fusiforme (H. fusiforme) via the ultrasonification extraction process were assessed in this study. Low molecular weight fucoidan improved the growth of human B and T cells, up to approximately 40% as compared to the controls (untreated) and 30% for commercially available fucoidan (Mw 150 kDa). IL-6 and TNF-$\alpha$ were secreted from human B cells at levels of $7.8\times10^{-4}$ pg/mL and $7.2\times10^{-4}$ pg/mL, respectively, and these levels were higher than the levels measured in the controls and with other high molecular weight fucoidan. It was also determined that the cytokine from human B and T cells cultivated with added fucoidan enhanced the growth of human NK cells. The fucoidan isolated from H. fusiforme showed low cytotoxicity, approximately 19%, after the addition of 1.0 mg/mL, the highest tested concentration. The growth of human lung cancer cells (A549) and human breast cancer cells (MCF-7) were inhibited by 69.8% and 83.3%, respectively. These results demonstrated that the low molecular weight fucoidan isolated from H. fusiforme has potential as a new functional food component that evidences immunomodulatory activities and anticancer activity. One of the primary positive features of this fucoidan is that low molecular weight polysaccharides can be readily handled during processing.

Epidemiology of central venous catheter related blood stream infections in pediatric patients (중심정맥 도관 관련 감염의 역학 고찰)

  • Kim, Jung Hyun;Eun, Ho Sun;Choi, Kyung Min;Kim, Dong Soo;Young, Dong Eun
    • Clinical and Experimental Pediatrics
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    • v.49 no.2
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    • pp.157-161
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    • 2006
  • Purpose : The purpose of this study is to investigate the pathogens of central venous catheter-related blood stream infections and search for the association among the insertion site, the duration and the underlying conditions with the prevalence of central venous catheter-related blood stream infections under 15 years old. Methods : A retrospective study was performed from Jan, 2003 to Dec, 2003 in Severance Hospital on 112 patients who undertook central venous catheter insertions. Results : We examined 112 patients who undertook central venous catheter insertion. The mean age of patients was $4.77{\pm}4.12$ years old. Coagulase negative Staphylococci was the most common organism of central venous catheter-related blood stream infections accounting for 33.9 percent, followed by Eenterococcus faecium(9.3 percent), and Staphylococcus aureus(7.5 percent), The most common insertion site was the right femoral vein, followed by the right jugular vein and the left femoral vein. The mean insertion period was $14.17{\pm}12.00$ days. Conclusion : Central venous catheter-related blood stream infections were not only related to the underlying conditions, but also to the insertion site. We need to study the clinical importance of coagulase negative Staphylococci as it is part of the normal flora of the skin. In future, more studies are needed to take preventive measures and improve treatment methods.

STUDY OF RAT EPIGASTRIC VESSELS ACCORDING TO THE FREEZING TIME : HISTOLOGIC, HISTOMORPHOMETRIC, IMMUNOHISTOCHEMICAL & SCANNING ELECTRON MICROSCOPIC STUDY (백서 상복부 혈관의 동결시간에 따른 변화에 대한 연구)

  • Kim, Woo-Chan;Lee, Chong-Heon;Kim, Kyung-Wook;Kim, Chang-Jin
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.21 no.2
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    • pp.89-109
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    • 1999
  • Vascular spasm which has been reported to occur in 25% of clinical cases continues to be a problem in microvascular surgery; When prolonged and not corrected, it can lead to low flow, thrombosis, and replant or free flap failure. Ischemia, intimal damage, acidosis and hypovolemia have been implicated as contributors to the vascular spasm. Although much work has been done on the etiology and prevention of vasospasm, a spasmolytic agent capable of firmly protecting against or reversing vasospasm has not been found. Therefore vascular freezing was introduced as a new safe method that immediately and permanently relieves the vasospasm and can be applied to microsurgical transfers. Cryosurgery can be defined as the deliberate destruction of diseased tissue or relief the vascular spasm in microvascular surgery by freezing in a controlled manner. 96 Sprague Dawley rats each weighing within 250g were used and divided into 2 group, experimental 1 and 2 group. In the experimental 1 group, right epigastric vessels (artery and vein) were freezed with a cryoprobe using $N_2O$ gas for 1 min. In the experimental 2 group, after freezing for 1 min, thawing for 30 secs and repeat freezing for 30 secs. Left side was chosen as control group in both group. We sacrified the experimental animals by 1 day, 3 days, 1 week, 2 weeks, 4 weeks & 5 months and observed the sequential change that occur during regeneration of epigastric vessels using a histologic, histomorphometric, immunohistochemical and SEM study after the vascular freezing. The results were as follows1. In epigastric arteries, internal diameters had statistically significant enlargement in 1 day, 3 days of Exp-1 group and 1 day, 3 days, 1 week & 2 weeks of Exp-2 group. Wall thickness had statistically significant thinning in 2 weeks of Exp-2 group. 2. In epigastric veins, internal diameters had enlargement of statistical significance in 1 day of Exp-1 and Exp-2 group. 3. The positive PCNA reactions in smooth muscle appeared in 1 week and increased until 2 weeks, decreased in 4 weeks. There was no statistical significance between Exp-1 and Exp-2 group. 4. The positive ${\alpha}$-SMA reaction in smooth muscles showed weak responses until 1 week and slowly increased in 2 weeks and showed almost control level in 4 weeks. 5. The positive S-100 reactions in the perivascular nerve bundles showed markedly decrease in 1 day, 3 days and increased after 1 week and showed almost control level in 4 weeks. Exp-1 group had stronger response than Exp-2 group. 6. In SEM, we observed defoliation of endothelial cell and flattening of vessel wall. Exp-2 group is more destroyed and healing was slower than Exp-1 group. To sum up, relief of vasospasm (vasodilatation) by freezing with cryoprobe was originated from the damage of smooth muscle layer and perivascular nerve bundle and the enlargement of internal diameter in vessels was similar to expeimental groups, but Exp-2 group had slower healing course and therefore vessel freezing in microsurgery can be clinically used, but repeat freezing time needs to be studied further.

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The Learning Curve of Laparoscopy-assisted Distal Gastrectomy (LADG) for Cancer (학습곡선을 기준으로 한 복강경 보조 원위절제술에 대한 결과)

  • Kim, Kab-Choong;Yook, Jeong-Hwan;Choi, Ji-Eun;Cheong, Oh;Lim, Jeong-Taek;Oh, Sung-Tae;Kim, Byung-Sik
    • Journal of Gastric Cancer
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    • v.8 no.4
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    • pp.232-236
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    • 2008
  • Purpose: Laparoscopic surgery for gastric cancer was introduced in the past decade because it was considered less invasive than open surgery, and this results in less postoperative pain, faster recovery and an improved quality of life. Several studies have demonstrated the safety and feasibility of this procedure. We examined the outcome of performing laparoscopic surgery for gastric cancer over the last two year. Materials and Methods: From April 2004 to December 2006, 329 patients with gastric adenocarcinoma underwent a laparoscopy-assisted distal gastrectomy with lymph node dissection. The data was retrospectively reviewed in terms of the clinicopathologic findings, the perioperative outcomes and the complications. Results: The total patient group was comprised 196 men (59.6%) and 133 women (40.4%). The mean BMI was 23.6 and the mean tumor size was 2.7 cm. The mean number of harvested lymph node was 22.7, and this was 18.6 before 30 cases and 23.1 after 30 cases, and the difference was significant (P=0.02). The mean operation time was 180.9 min, and this was than 287.9 min before 30 cases and 170.2 min after 30 cases. After 30 cases, there was a significant improvement of the operation time (P<0.01). The mean incision length after 30 cases was shorter than that before 30 cases (P<0.01). Postoperative complications occurred in 24 (7.3%) of 329 patients and there was no conversion to open surgery. Conclusion: Even though the LADG was accompanied by a difficult learning curve, we successfully performed 329 LADG procedures over the past 2 years and we believe that LADG is a safe, feasible operation for treating most early gastric cancers (EGC).

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