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Genomic analysis of Mycobacterium fortuitum by pulsed-field gel electrophoresis (Pulsed-field Gel Electrophoresis를 이용한 Mycobacterium fortuitum의 유전형 분석)

  • Lee, Tae-Yoon;Do, In-A;Kim, Sung-Kwang
    • Journal of Yeungnam Medical Science
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    • v.12 no.2
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    • pp.366-385
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    • 1995
  • Epidemiological studies are important in both the prevention and treatment of mycobacterial infections. This study was initiated to establish the pulsed-field gel electrophoresis (PFGE) method, which are not yet extensively studied. The most apprpriate restriction endonucleases included DraI, AsnI, and XbaI. The optimal PFGE condition was different according to the enzymes used. Two stage PFGE was performed, in case of DraI first stage was performed with 10 seconds of initial pulse and 15 seconds of final pulse, while the second stage was performed with 60 seconds of initial pulse and 70 seconds of final pulse. The electrophoresis time for DraI-PFGE was 14 hours for each stage. Electrophoresis was performed for 22 hours, in case of XbaI, with 3 seconds of initial pulse and 12 seconds of final pulse. Electrophoresis was performed for 22 hours, in case of AsnI, with 5 seconds of initial pulse and 25 seconds of final pulse. In all cases the voltage of the electrophoresis was maintained constantly at 200 voltage. Standard mycobacterial strains, which included Mycobacterium bovis BCG, M. tuberculosis, and M. fortuitum, could not be differentiated by PFGE analysis. PFGE analysis was performed to differentiate 9 clinically isolated M. fortuitum strains using AsnI. All M. fortuitum strains showed different genotypes except 2 strains. Cluster analysis divided M. fortuitum strains into 2 large groups. PFGE analysis was performed to further differentiate M. fortuitum isolates using XbaI. The undifferentiated 2 M. fortuitum strains showed different PFGE patterns with Xba I. Cluster analysis of the XbaI-PFGE patterns showed more complex grouping than AsnI-PFGE patterns, which showed that XbaI-PFGE analysis was better than AsnI-PFGE in M. fortuitum genotyping. The top dissimilarity values of AsnI-PFGE and XbaI-PFGE were 0.74 and 0.75, respectively. This value was higher than that of arbitrarily primed polymerase chain reaction (AP-PCR) analysis and lower than that of restriction fragment length polymorphism (RFLP) analysis. This suggested that PFGE can be used as a supportive or alternative genotyping method to RFLP analysis.

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Manufacturing and Characteristics of Fruit Wine from Acanthapanax sessiliflorus (오가피 열매 발효주의 제조 및 특성)

  • Choi, Jae-Myoung;Kim, Kwang-Yup;Lee, Sang-Hwa;Ahn, Jun-Bae
    • Food Engineering Progress
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    • v.14 no.1
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    • pp.1-6
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    • 2010
  • Fruit wine from Acanthopanax sessiliflorus(A. sessiliflorus) including many pharmacological components was manufactured and its functional properties were investigated. The fruit part of A. sessiliflorus contained 75.74${\pm}$0.49%(w/w) moisture, 12.51${\pm}$1.23%(w/w) crude protein, 4.20${\pm}$0.51.%(w/w) crude fat and 5.21${\pm}$1.64%(w/w) crude ash. Minerals of fruit were potassium(12.94${\pm}$0.08 mg/g), calcium(1.53${\pm}$0.06 mg/g) and magnesium(1.12${\pm}$0.05 mg/g). Initial soluble solid and fermentation temperature were 24-30$^{\circ}$Brix and 20${^{\circ}C}$ for manufacturing fruit wine from A. sessiliflorus. When initial soluble solid of a must was adjusted to more than 30$^{\circ}$Brix, ethanol production was suppressed slightly. The polyphenol content of the fruit wine fermented at 20${^{\circ}C}$(125.24${\pm}$1.86 mg/mL) was higher than those at 25${^{\circ}C}$(99.69${\pm}$2.11 mg/mL) and 30${^{\circ}C}$(95.55${\pm}$1.54 mg/mL). Electron donating activities of wines fermented at 20, 25 and 30${^{\circ}C}$ were 85.9${\pm}$2.3, 55.7${\pm}$2.5 and 55.2${\pm}$3.4%, respectively. The content of eleutheroside B increased up to 146.58${\pm}$4.10 $\mu$g/mL during fermentation. There was no significant effect of fermentation temperature on eleutheroside B content. The fruit part of A. sessiliflorus can be used as a valuable resource for development of nutraceutical foods.

Induction of Apoptosis by Water Extract of Glycyrrhizae radix in Human Bladder T24 Cancer Cells (인체 방광암 T24 세포에서 감초(Glycyrrhizae radix) 열수추출물에 의한 apoptosis 유도)

  • Lee, Ki Won;Kim, Jeong Il;Lee, Seung Young;Choi, Kyung-Min;Oh, Young Taek;Jeong, Jin-Woo
    • Korean Journal of Plant Resources
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    • v.32 no.4
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    • pp.255-263
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    • 2019
  • Glycyrrhizae radix is one of the most frequently prescribed ingredients in Oriental medicine, and Glycyrrhizae radix extract has been shown to exert anti-cancer effects. However, the cellular and molecular mechanisms of programed cell death (apoptosis) by Glycyrrhizae radix are poorly defined. In the present study, it was examined the molecular mechanisms of apoptosis by water extracts of Glycyrrhizae radix (GRW) in human bladder T24 cancer cells. It was found that GRW could inhibit the cell growth of T24 cells in a concentration-dependent manner, which was associated with the induction of apoptotic cell death, as evidenced by the formation of apoptotic bodies, DNA fragmentation and increased populations of annexin-V positive cells. The induction of apoptotic cell death by GRW was connected with an up-regulation of pro-apoptotic Bax protein expression and down-regulation of anti-apoptotic proteins (Bcl-2 and Bcl-xL), and inhibition of apoptosis family proteins (XIAP, cIAP-1 and cIAP-2). In addition, apoptosis-inducing concentrations of GRW induced the activation of caspase-9, an initiator caspase of the mitochondrial-mediated intrinsic pathway, and caspase-3, accompanied by proteolytic degradation of PARP. GRW also induced apoptosis via a death receptor-mediated extrinsic pathway by caspase-8 activation, resulting in the down-regulation of total Bid and suggesting the existence of cross-talk between the extrinsic and intrinsic pathways. Taken together, the present results suggest that GRW may be a potential chemotherapeutic agent for the control of human bladder cancer cells.

Influences of the devastated forest lands on flood damages (Observed at Chonbo and the neighbouring Mt. Jook-yop area) (황폐임야(荒廢林野)가 수해참상(水害慘狀)에 미치는 영향(影響) (천보산(天寶山)과 인접(隣接) 죽엽산(竹葉山)을 중심(中心)으로))

  • Chung, In Koo
    • Journal of Korean Society of Forest Science
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    • v.5 no.1
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    • pp.4-9
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    • 1966
  • 1. On 13 September 1964 a storm raged for 3 hours and 20 minutes with pounding heavy rainfalls, and precipitation of 287.5 mm was recorded on that day. The numerous landslides were occured in the eroded forest land neighbouring Mt. Chunbo, while no landslides recorde at all on Mt. Jookyup within the premise of Kwangnung Experiment Station, the Forest Experiment Station. 2. Small-scalled Landslides were occured in 43 different places of watershed area (21.97 ha.) in which the survey had already been done, in and around Mt. Chunbo (378 m a.s.l.). The accumulated soil amount totaled $2,146,56m^3$ due to the above mentioned landslides, while soil accumulated from riverside erosion has reached to $24,168.79m^3$, consisting of soils, stones, and pebbles. However, no landslides were reported in the Mt. Jook yup area because of dense forest covers. The ratio of the eroded soil amount accumulated from the riversides to that of watershed area was 1 to 25. On the other hand, the loss and damage in the research area of Mt. Chonbo are as follows: 28 houses completly destroyed or missing 7 houses partially destroyed 51 men were dead 5 missing, and 57 wounded. It was a terrible human disaster However, no human casualties were recorded at all, 1 house-completly destroyed and missing, 2 houses-partially destroyed. Total:3 houses were destroyed or damaged, in The area of Mt. Jookyup 3. In the calculation of the quanty of accumulated soil, the or mula of "V=1/3h ($a+{\sqrt{ab}}+b$)" was used and it showed that 24, 168.79m of soil, sands, stones and pebbles carried away. 4. Average slope of the stream stood 15 at the time of accident and well found that there was a correlation between the 87% of cross-area sufferd valley erosion and the length of eroded valley, after a study on regression and correlation of the length and cross-area. In other works, the soil erosion was and severe as we approached to the down-stream, counting at a place of average ($15^{\circ}1^{\prime}$) and below. We might draw a correlation such as "Y=ax-b" in terms of the length and cross-area of the eroded valley. 5. Sites of char-coal pits were found in the upper part of the desert-like Mt. Chunbo and a professional opinion shows that the mountain was once covered by the oak three species. Furthermore, we found that the soil of both mountains have been kept the same soil system according to a research of the soil cross-area. In other words, we can draw out the fact that, originally, the forest type and soil type of both Mt. Chunbo (378m) and Mt. Jookyup (610m) have been and are the same. However, Mt. Chunbo has been much more devastated than Mt. Jookyup, and carried away its soil nutrition to the extent that the ratios of N. $P_2O_5K_2O$ and Humus C.E.C between these two mountains are 1:10;1:5 respectively. 6. Mt. Chunbo has been mostly eroded for the past 30 years, and it consists of gravels of 2mm or larger size in the upper part of the mountain, while in the lower foot part, the sandy loam was formulated due to the fact that the gluey soil has been carried and accumulated. On the hand, Mt. Jookyup has consitantly kept the all the same forest type and sandy loam of brown colour both in the upper and lower parts. 7. As for the capability of absorbing and saturating maximum humidity by the surface soil, the ratios of wet soil to dry soil are 42.8% in the hill side and lower part of the eroded Mt. Chunbo and 28.5% in the upper part. On the contrary, Mt. Jookyup on which the forest type has not been changed, shows that the ratio in 77.4% in the hill-side and 68.2% in the upper part, approximately twice as much humidity as Mt. Chunbo. This proves the fact that the forest lands with dense forest covers are much more capable of maintaining water by wood, vegitation, and an organic material. The strength of dreventing from carring away surface soil is great due to the vigorous network of the root systems. 8. As mentioned above, the devastated forest land cause not only much greater devastation, but also human loss and property damage. We must bear in mind that the eroded forest land has taken the valuable soil, which is the very existance of origin of both human being and all creatures. As for the prescription for preventing erosion of forest land, the trees for furtilization has to be planted in the hill,side with at least reasonable amount of aertilizer, in order to restore the strength of earth soil, while in the lower part, thorough erosion control and reforestation, and establishments along the riversides have to be made, so as to restore the forest type.

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Effect on Shear Strength of Ceramic Surface Treatment Materials and Three Resin Cements to IPS Empress 2 (표면 처리재와 레진 시멘트가 IPS Empress 2의 전단결합강도에 미치는 영향)

  • Yae, Sun-Hae;Lee, Kyubok;Lee, Cheong-Hee
    • Journal of Dental Rehabilitation and Applied Science
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    • v.18 no.3
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    • pp.157-170
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    • 2002
  • The purpose of this study is to analyze the shear bond strength according to kinds of surface treatment agents and resin cements after acid etching of IPS Empress 2. For this purpose, test groups were classified into silane-treated bonding groups, silica-coated group and Targis link applied group. Then, nine bonding groups in total, each three groups per kind, were prepared by using three kinds of resin cements-Panavia F, Variolink II and Rely-X ARC, and thirty test specimens per group were prepared. To examine any changes in the oral environment, the shear bond strength of each test specimen was measured after dipping test for 24 hours and for five weeks, respectively, in distilled water at $37^{\circ}C$ and performing heat cycle 10,000 times in total, each 2,000 times per week, during a five weeks of dipping, under the condition similar to the oral environment. The bond failure modes were also observed by means of a scanning electron microscope. The results are summarized as follows 1. Statically significant differences between the surface conditioning materials were observed. The shear strength of the silane treatment was the highest of all three types of surface treatments(p<0.001). 2. Shear strengths varied significantly for different types of resin luting agents. But bond strength of Targis link surface treatments were not significantly different(p<0.05). 3. No significant difference of bonding strengths was found between storage time conditions(24 hours and 5 weeks). Only group II, IV, VII, IX were significantly different(p<0.05). 4. After thermocycling, the shear bond strengths of all groups were significantly decreased (p<0.05). Group III, V, VI were no significantly different. 5. On the SEM observation of fractured surfaces, all groups were shown complex failure.

Ionic currents elicited by the hypotonic solution in hamster eggs (저장성 용액에 노출된 햄스터 난자에 관찰되는 이온전류의 변화)

  • Choi, Won-yeong;Kim, Yang-mi;Haan, Jae-hee;Huh, Il-oh;Park, Choon-ok;Hong, Seong-geun;Pyu, Pan-dong;Kim, Jong-shu
    • Korean Journal of Veterinary Research
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    • v.36 no.2
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    • pp.305-312
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    • 1996
  • Cell volume regulatory mechanisms are usually disclosed by exposure of cell to anisotonic media. If a cell is suddenly exposed to hypotonic media, it swells initially like an osmometer but within minutes regains its original cell volume. This behavior has been labelled as regulatory cell volume decrease(RVD). RVD is believed to result from the loss of permeable ions through the membrane. In this study, we examined hypotonically induced changes in the membrance currents involved in RVD by using whole cell voltage clamp technique in the unfertilized hamster egg. At -40mV of the holding potential, the stationary current was maintained in the hamster egg exposed to isotonic solution composed of, mainly, 115mM NaCl and 40mM mannitol. Hypotonic solution was prepared by removing mannitol. Therefore, the concentrations of $Na^+$ and $Cl^-$ in this hypotonic media were the same as those in the isotonic solution. Following 30 to 60 sec after applying the hypotonic media to the egg, the inward current was evoked. This inward current was eliminated by $100{\mu}M$ 4-acetamido-4'-isothiocyanostil-bene-2,2'-disulfonic acid(SITS), an anion channel blocker, leaving the small outward current component. Further addition of 2mM $Ba^{2+}$, a broad $K^+$ channel blocker, completely abolished the small outward current left even in the presence of SITS during hypotonic stress. These results suggest that $K^+$ and $Cl^-$ move out of cells, resulting in RVD. To test the involvement of $Na^+$ in RVD, 20mM Na-isethionate was substituted for mannitol in isotonic media(135mM $Na^+$) and Na-isethionate (20mM) was freed the hypotonic solution. Only $Cl^-$ concentration in both isotonic and hypotonic media was kept constant at 115mM, whereas concentration of $Na^+$ was lowered in hypotonic solution to 115mM from 135mM in isotonic solution. Hypotonic medium induced the outward current in the egg equilibrated isotonically. This current was reduced by $100{\mu}M$ SITS but was augmented by 2 mM $Ba^{2+}$. In terms of RVD, these results imply that $Cl^-$ efflux is coupled with $K^+$, maybe for electroneutrality during hypotonic stress and/or with $Na^+$ via unknown transport mechanism(s). From the overall results, the hypotonic stress facilitates the movement of $Cl^-$ and $K^+$ out of the hamster egg to regain cellular volume with electroneutrality. If there exist a difference in $[Na^+]_0$ between isotonic and hypotonic solution, another transport mechanism concerned with $Na^+$ may, at least partly, participate in regulatory volume decrease.

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Quantitative Comparisons between CT and $^{68}Ge$ Transmission Attenuation Corrected $^{18}F-FDG$ PET Images: Measured Attenuation Correction vs. Segmented Attenuation Correction (CT와 $^{68}Ge$ 감쇠보정 $^{18}F-FDG$ PET 영상의 정량적 비교: 측정감쇠보정대 분할감쇠보정)

  • Choi, Joon-Young;Woo, Sang-Keun;Choi, Yong;Choe, Yearn-Seong;Lee, Kyung-Han;Kim, Byung-Tae
    • Nuclear Medicine and Molecular Imaging
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    • v.41 no.1
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    • pp.49-53
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    • 2007
  • Purpose: It was reported that CT-based measured attenuation correction (CT-MAC) produced radioactivity concentration values significantly higher than $^{68}Ge$-based segmented attenuation correction (Ge-SAC) in PET images. However, it was unknown whether the radioactivity concentration difference resulted from different sources (CT vs. Ge) or types (MAC vs. SAC) of attenuation correction (AC). We evaluated the influences of the source and type of AC on the radioactivity concentration differences between reconstructed PET images in normal subjects and patients. Material and Methods: Five normal subjects and 35 patients with a known or suspected cancer underwent $^{18}F-FDG$ PET/CT. In each subject, attenuation corrected PET images using OSEM algorithm (28 subsets, 2 iterations) were reconstructed by 4 methods: CT-MAC, CT-SAC, Ge-MAC, and Ge-SAC. The physiological uptake in normal subjects and pathological uptake in patients were quantitatively compared between the PET images according to the source and type of AC. Results: The SUVs of physiological uptake measured in CT-MAC PET images were significantly higher than other 3 differently corrected PET images. Maximum SUVs of the 145 foci with abnormal FDG uptake in CT-MAC images were significantly highest among 4 differently corrected PET images with a difference of 2.4% to 5.1% (p<0.001). The SUVs of pathological uptake in Ge-MAC images were significantly higher than those in CT-SAC and Ge-MAC PET images (p<0.001). Conclusion: Quantitative radioactivity values were highest in CT-MAC PET images. The adoption of MAC may make a more contribution than the adoption of CT attenuation map to such differences.

Peritoneal Protein Loss in Nephrotic Syndrome on Peritoneal Dialysis (복막 투석 중인 신증후군 환자의 복막을 통한 단백 소실)

  • Ahn, Yo-Han;Jung, Eui-Seok;Lee, Se-Eun;Lee, Hyun-Gyung;Lee, So-Hee;Kang, Hee-Gyung;Ha, Il-Soo;Jung, Hae-Il;Choi, Yong
    • Childhood Kidney Diseases
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    • v.13 no.2
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    • pp.189-196
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    • 2009
  • Purpose : The pathophysiologic mechanism of nephrotic syndrome is not yet known clearly. At least in some cases, certain 'circulating factors' are thought to increase the glomerular protein permeability. Considering the systemic effect of the circulating factor on peritoneal membrane, we evaluated the loss of protein through peritoneal membrane in patients on peritoneal dialysis due to the end stage renal disease (ESRD) caused by steroid resistant nephrotic syndrome (SRNS). Methods : We retrospectively reviewed the medical records of 26 pediatric patients on peritoneal dialysis ensued during the period from 2001 to 2007 at our clinic. Twelve patients had SRNS, while 14 patients had ESRD caused by the congenital anomalies of urinary system. Results : While the other parameters including nPNA indicating the adequacy of protein intake were similar between the two groups, serum albumin was lower in SRNS patients than the non-SRNS patients ($3.7{\pm}0.3$ g/dL vs. $4.0{\pm}0.4$ g/dL, P=0.021). Peritoneal protein loss was higher in SRNS patients than in non-SRNS patients ($3,044.4{\pm}837.6\;mg/m^2$/day vs. $1,791.6{\pm}1,244.0\;mg/m^2$/day, P=0.007). The protein permeability of the peritoneal membrane measured by the ratio of total protein concentration in dialysate to plasma was twice as high in SRNS patients as the non-SRNS ($1.06{\pm}0.46%$ vs. $0.58{\pm}0.43%$, P=0.010). After 1 year, peritoneal protein loss increased in both patient groups, but to a significantly greater degree in non-SRNS patient (P=0.023). Conclusion : The results of our study support the notion that in nephrotic syndrome there are some 'circulating factors' with the systemic effect. Since the greater protein loss through peritoneal membrane in SRNS was confirmed in this study, more meticulous nutritional support and close monitoring on the nutrition are required in these patients.

Changes in the Functional Components of Lactobacillus acidophilus-Fermented Red Ginseng Extract and Its Application to Fresh Cheese Production (Lactobacillus acidophilus로 발효한 홍삼 농축액의 기능성 성분 변화 및 이를 이용한 신선치즈 제조)

  • Park, Jong-Hyuk;Moon, Hye-Jung;Oh, Jeon-Hui;Lee, Joo-Hee;Jung, Hoo-Kil;Choi, Kyung-Min;Cha, Jeong-Dan;Lim, Ji-Ye;Han, Su-Beom;Lee, Tae-Bum;Lee, Min-Jung;Choi, Hye-Ran
    • Journal of Dairy Science and Biotechnology
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    • v.32 no.1
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    • pp.47-53
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    • 2014
  • In this study, our aim was to investigate the changes in ginsenosides and polyphenols in red ginseng extract fermented by Lactobacillus acidophilus and to manufacture fresh cheese using fermented red ginseng extract. Red ginseng extract (3%, w/v) was fermented by L. acidophilus for 24 h. On performing lactic acid bacteria counts, we determined that L. acidophilus reached its maximum growth phase after 16 h; this was followed by decrease in growth. During fermentation, the levels of ginsenosides Rg3 (20S) and Rg3 (20R) as well as protopanaxadiol (20R), F1, and compound K increased, while those of s Rb2, Rd, Rf, and Rg1 decreased. The pH, titratable acidity, and viable cell counts in fresh cheese prepared using fermented red ginseng extract were measured during the storage period. The pH decreased over time, while titratable acidity and viable cell counts increased with increase in the duration of the storage period. Sensory tests showed that the overall sensory properties of fresh cheese prepared using 1% fermented red ginseng extract were similar to those of the control groups. This result suggests that L. acidophilus-fermented red ginseng has potential for development as a new bioactive material.

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Evaluation on Usefulness of Applying Body-fix to Liver Cancer Patient in Tomotherapy (간암환자의 토모치료시 Body-fix 사용유무에 따른 유용성 평가)

  • Oh, Byeong-Cheon;Choi, Tae-Gu;Kim, Gi-Chul
    • The Journal of Korean Society for Radiation Therapy
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    • v.22 no.1
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    • pp.11-18
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    • 2010
  • Purpose: In every time radiation therapy set up errors occur because internal anatomical organs move due to breathing and change of patient's position. These errors may affect the change of dose distribution between target area and normal structure. This study investigates the usefulness of body-fix in clinical treatment. Materials and Methods: Among 55~60 aged male patients who has hepatocellular carcinoma in area of liver's couinaud classification, we chose 10 patients and divided two groups by using body-fix or not. When applying body-fix, we maintained a vacuum of 80 mbar pressure by using vacuum pump (Medical intelligence, Germany). Patients had free breathing with supine position. After working to fuse and consist MV-CT (megavoltage computed tomography) with KV-CT (kilovoltage computed tomography) obtained by 5 times treatments, we compared and analyzed set up errors occurred to (Right to Left, RL) of X axis, (Anterioposterio, AP) of Z axis, (Cranicoudal, CC) of Y axis. Results: Average Set up errors through image fusion showed that group A moved $0.3{\pm}1.1\;mm$ (Cranicoudal, CC), $-1.1{\pm}0.7\;mm$ (Right to Left, RL), $-0.2{\pm}0.7\;mm$ (Anterioposterio, AP) and group B moved $0.62{\pm}1.94\;mm$ (Cranicoudal, CC), $-3.62{\pm}1.5\;mm$ (Right to Left, RL), $-0.22{\pm}1.2\;mm$ (Anterioposterio, AP). Deviations of X, Y and Z axis directions by applying body-fix indicated that maximum X axis was 5.5 mm, Y axis was 19.8 mm and Z axis was 3.2 mm. In relation to analysis of error directions, consistency doesn't exist for every patient but by using body-fix showed that the result of stable aspect in spite of changes of everyday's patient position and breathing. Conclusion: Using body-fix for liver cancer patient is considered effectively for tomotherapy. Because deviations between group A and B exist but they were stable and regular.

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