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Establishment and Characterization of Bone Marrow Mesenchymal Stromal/Stem Cells (MSCs) Derived from ${\alpha}$-1,3-Galactosyltransferase Knock Out(GalT KO) Pig (${\alpha}$-1,3-Galactosyltransferase Knock Out(GalT KO) 돼지유래 골수 중간엽 줄기세포의 특성 규명)

  • Ock, Sun-A;Oh, Keon Bong;Hwang, Seongsoo;Im, Seoki;Kim, Youngim;Park, Jin-Ki
    • Journal of Embryo Transfer
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    • v.28 no.3
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    • pp.281-287
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    • 2013
  • A major barrier to progress in pig to primate organ transplantation or cell therapy is the presence of terminal ${\alpha}$-1,3-galactosyl epitopes on the surface of pig cells. Therefore, the purpose of this experiment was to establish and cha- racterize mesenchymal stromal/stem cells (MSCs) derived from ${\alpha}$-1,3-galactosyltransferase (GalT) knock out (GalT KO) pig to confirm their potential for cell therapy. Bone marrow (BM)-MSCs from GalT KO pig of 1 month old were isolated by Ficoll-Paque PLUS gradient and cultured with A-DMEM + 10% FBS on plastic dishes in 5% $CO_2$ incubator at 38.5. GalT KO BM-MSCs were analyzed for the expression of CD markers ($CD45^-$, $29^+$, $90^+$ and $105^+$) and in vitro differentiation ability (adiopogenesis and osteogenesis). Further, cell proliferation capacity and cell aging of GalT KO BM-MSCs were compared to Wild BM-MSCs by BrdU incorporation assay (Roche, Germany) using ELISA at intervals of two days for 7 days. Finally, the cell size was also evaluated in GalT KO and Wild BM-MSCs. Statistical analysis was performed by T-test (P<0.05). GalT KO BM-MSCs showed fibroblast-like cell morphology on plastic culture dish at passage 1 and exhibited $CD45^-$, $29^+$, $90^+$ and $105^+$ expression profile. Follow in ginduction in StemPro adipogenesis and osteogenesis media for 3 weeks, GalT KO BM-MSCs were differentiated into adipocytes, as demonstrated by Oilred Ostaining of lipid vacuoles and osteocytes, as confirmed by Alizarinred Sstaining of mineral dispositions, respectively. BrdU incorporation assay showed a significant decrease in cell proliferation capacity of GalT KO BM-MSCs compared to Wild BM-MSCs from 3 day, when they were seeded at $1{\times}10^3$ cells/well in 96-well plate. Passage 3 GalT KO and Wild BM-MSCs at 80% confluence in culture dish were allowed to form single cells to calculate cell size. The results showed that GalT KO BM-MSCs($15.0{\pm}0.4{\mu}m$) had a little larger cell size than Wild BM-MSCs ($13.5{\pm}0.3{\mu}m$). From the above findings, it is summarized that GalT KO BM-MSCs possessed similar biological properties with Wild BM-MSCs, but exhibited a weak cell proliferation ability and resistance to cell aging. Therefore, GalT KO BM-MSCs might form a good source for cell therapy after due consideration to low proliferation potency in vitro.

The Anti-inflammatory Effect of Skipjack Tuna (Katsuwonus pelamis) Oil in LPS-induced RAW 264.7 Cells and Mouse Models (LPS 유도 RAW 264.7 세포와 마우스 모델에서 참치(Katsuwonus pelamis) 유의 항염증 효과)

  • Kang, Bo-Kyeong;Kim, Min-Ji;Kim, Koth-Bong-Woo-Ri;Ahn, Na-Kyung;Choi, Yeon-Uk;Bark, Si-Woo;Pak, Won-Min;Kim, Bo-Ram;Park, Ji-Hye;Bae, Nan-Young;Ahn, Dong-Hyun
    • Microbiology and Biotechnology Letters
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    • v.43 no.1
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    • pp.45-55
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    • 2015
  • This study was carried out to demonstrate the anti-inflammatory effect of tuna oil (TO) using LPS-induced inflammation responses and mouse models. First, nitric oxide (NO) and pro-inflammatory cytokines levels were suppressed up to 50% with increasing concentrations of TO without causing any cytotoxicity. Also, the expression of a variety of proteins, such as inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) and nuclear factor kappa B (NF-κB), was suppressed in a dosedependent manner by treatment with TO. Furthermore, TO also inhibited the phosphorylation of mitogen-activated protein kinases (MAPKs), including c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase (ERK), and p38 protein kinase (p38). Moreover, in in vivo testing the formation of ear edema was reduced at the highest dose tested compared to that in the control, and a reduction of ear thickness and the number of mast cells was observed in histological analysis. In acute toxicity test, no mortalities occurred in mice administrated 5,000 mg/kg body weight of TO over a two-week observation period. Our results suggest that TO has a considerable anti-inflammatory property through the suppression of inflammatory mediator productions and that it could prove to be useful as a potential anti-inflammatory therapeutic material.

Retrospective Cohort Study on the Administration of Sedative for Delirium in Terminally Ill Cancer Patients and Survival Time (말기암환자의 섬망으로 인한 진정제 투약과 생존기간에 관한 후향적 코호트 연구)

  • Park, Hyoung Sook;Kim, Dae Sook;Bae, Eun Hee;Kim, Jung Rim;Seo, Jung Hwa;Yun, Jung Mi
    • Journal of Hospice and Palliative Care
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    • v.19 no.2
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    • pp.119-126
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    • 2016
  • Purpose: This study analyzed the difference in survival time of patients with delirium according to sedative medication. Methods: From January 2012 through December 2013, a retrospective cohort study was performed using the electronic medical records (EMR) of Pusan National University Hospital. Among 900 patients who died from cancer, we selected 240 who suffered delirium based on the EMR. The Nu-DESC delirium screening test was used to diagnose delirium. Results: The median length of delirium period was five days. Delirium characteristics were dominated by inappropriate behaviors (35.0%). Sedatives were administered in 72.1% of the cases. The most frequently used sedative was haloperidol which was used in 59.6% of cases. The delirium period significantly differed by patients' age (F=3.96, P=0.021), cancer type (F=3.31, P=0.010), chemotherapy (t=-3.44 P=0.001). The average survival time was 16.85 days for the sedative medication group and 9.37 days for the non-medication group, which, however, was not significant (t=1.766, P=0.079). Conclusion: In this study, the use of sedatives did not affect patients' survival time. Thus, appropriate sedative medication can be positively recommended to comfort terminal cancer patients and their families.

Cross-Protection Effectiveness of Cucumber mosaic virus (CMV) Isolates Associated with Satellite RNA for Prevention of CMV Disease in Pepper Plants (Satellite RNA 보유 Cucumber mosaic virus(CMV)의 고추 CMV병에 대한 교차방어 효과)

  • 최장경;성미영;정혜진;홍진성;이상용
    • Research in Plant Disease
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    • v.7 no.3
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    • pp.155-163
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    • 2001
  • Two attenuated Cucumber mosaic virus (CMV) isolates, Paf-CMV and Rs2-CMV that had been selected from CMV isolates associated with satellite RNA (satRNA) were tested for cross-protection effect in pepper plants. The viruses selected as attenuated strains appeared to be identical serologically and physically to the challenge virus (Mf-CMV), but they were lower in the dilution end-point of infectivity of crude sap than Mf-CMV When symptoms were observed in several indicator plants after inoculation, Paf-CMV and Rs2-CMV were symptomless or showed mild mosaic symptoms while another satRNA isolate Ap-CMV developed severe mosaic symptoms on the leaves as Mf-CMV. The nucleotide sequences of the satRNAs were determined by sequencing full-length cDNA clones. Paf-, Rs2- and Ap-satRNAs were 386, 335, and 347 nucleotides long, respectively, The sequences were then compared with the other known Y-satRNA, revealing that nucleotide sequences of the satRNAs consisted of 5'- and 3'-terminal conserved regions. However variations occurred on the middle regions of the sequences, especially those related to symptom interference, showing significant differences between Paf-satRNA and other isolates. Infectious transcripts of Paf-satRNA and Rs2-satRNA induced mild mosaic symptoms in pepper plants when supported by genomic RNAs of Mf-CMV. Under greenhouse conditions, Paf-CMV and Rs2-CMV were tested for cross-protection effect in pepper and tobacco (Nicotiana tabacum cv, Xanthi nc) plants against Mf-CMV. No symptoms were developed on the plants vaccinated with Paf-CMV until 3 weeks after inoculation with the virulent strain; however another attenuated isolate, Rs2-CMV, showed less effectiveness in cross-protection. Depending on the concentration of the challenged virus, symptoms sometimes appeared later in the upper leaves. However, in plants challenged with low concentrations (below 0.2 mg/ml) of the challenge inoculum, symptoms caused by the virulent strain did not develop on the plants vaccinated with Paf-CMV. In the field experiments, the number of pepper plants with severe mosaic symptoms in the control plots was progressively increased after transplanting and reached approximately 50% after 50 days. On the other hand, the incidence of mosaic disease appeared very low on the plants that had received the protective inoculation with Paf-CMV.

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The Need for Child Hospice Care in Families of Children with Cancer (암 환아 가족의 아동 호스피스 요구도)

  • Kang, Kyung-Ah;Kim, Shin-Jeong;Kim, Young-Soon
    • Journal of Hospice and Palliative Care
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    • v.7 no.2
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    • pp.221-231
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    • 2004
  • Purpose: The purpose of this study was to analyze the need for child hospice care programs in families of children with cancer. Methods: The survey of 104 families who were taking care of children with cancer was conducted. This survey was conducted from February 2004 to July 2004 at two general hospitals in Seoul. The data were collected through a self-reporting questionnaire of 22 items. The items were classified into five areas by factor analysis to identify the construct validity. The reliability of the tool was established by Cronbach's alpha as .94 and the data collected were analyzed by descriptive statistics, t-test and ANOVA. Results: 1) The degree of need for hospice care of the subjects showed a high average of 3.40 (${\pm}3.8$). The need for 'emotional care of children' showed the highest mean (M=3.55), 'management of terminal physical symptoms'(M=3.49), 'control of secondary physical problems' (M=3.41), 'acceptance of the family's difficulty' (M=3.20), 'spiritual care for preparing for death'(M=3.17), respectively. 2) With respect to the demographic characteristics of the subjects, there were statistically significant differences in hospice care needs, according to the child's mother's age (F==4.980, P=.009), whether or not there were cancer patients among their siblings or relatives (t=2.423, P=.017). Conclusion: The family of children with cancer have a heavy burden of ambivalence, especially in relieving the anxiety and fear of their children, communicating about death, and managing physical symptoms. Child hospice care must be provided considering the needs of families of children with cancer. Thus popular needs as well as hospice nurses' higher concern and support for hospice care of children require further education and program development to meet the current demands.

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의료인의 호스피스가정간호에 대한 지식과 태도 조사연구

  • Kim, Ok-Gyeom
    • Korean Journal of Hospice Care
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    • v.2 no.2
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    • pp.28-48
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    • 2002
  • The advances of medical technologies have not only prolonged human life span, but also extended suffering period for the patients with incurable medical diseases. Hospice movement was developed to help these patients keep dignity and lives peaceful at the end of their life. Since many patients prefer to spend the last moment of life at home with their family, hospice home care has become very popular worldwide. The purpose of this study for a promotion and development of hospice home care in Korea, and features basic research on medical profession's knowledge and attitudes to hospice home care. This study which was used for the research questionnaires developed by the researcher that were answered by 100 physicians and 127 nurses in a general hospital. Data were collected from April 22, 2002 to May 10, 2002. The SPSS was used to make a comparative analysis of the frequency, percentile, ANOVA, and x2-test. The results of the study were as follows; 1.The medical profession showed high level of knowledge of the definition and philosophy of hospice. However, the physician group of the examinees showed insufficient knowledge of the fact that hospice care includes bereavement care, while the nurse group's response to the same question showed a significant difference(x2=10.752, p=.001). 2.For whom the hospice home care is provided, 95.6% of the respondents showed very high level of knowledge as answering that the incurable terminal illness patients and their families are the beneficiaries of hospice care. The respondents counted nurses, volunteers, pastors, physicians and social workers, consecutively, as hospice care providers. More nurse were positive toward pastors than physicians in regarding as a hospice care provider by a significant difference(x2=11.634, p=.001). 3.For when to referral hospice home care to the patients, only 34.2% answered that patients with less than 6 months of survival time are advised to receive hospice care, reflecting very low level of knowledge. 23.0% of the physicians and 48.0% of the nurses answered that hospice care should be provided when death is imminent, making a significant difference between the two groups(x2=6.413, p=.000). 4.To promote hospice activities, 87.2% pointed out that it is crucial to make general people, including those engaging in the medical field, more aware of hospice. 79.7% answered that a national hospice management should be developed, marking a significant difference between the physician group and nurse group(x2=10.485, p=.001). 5.Advantages of hospice home care are 87.2% responded that patients can have better rest at home receiving hospice home care. Economical merit was brought forward as one of the advantages also, where there was a significant difference between the physicians group and nurse group(x2=7.009, p=.008). 6.The medical professions' attitude to hospice home care are 92.8% of the physicians answered that they would advise incurable terminally ill patients to be discharged from hospital, with 44.3% of them advising the patients to receive hospice home care after leaving the hospital. From the nurses' point of view, 20.9% of the terminally ill patients are being referred to hospice home care after discharge, which makes a significant difference from the physicians' response(x2=19.121, p=.001). 7. 30.6% of physicians have referred terminally ill patients to hospice home care, 75.9% of whom were satisfied with their decision. Those physicians who have never referred their patients to hospice home care either did not know how to do it(66.7%) or were afraid of losing trust by giving the patients an impression of giving up(27.3%). 94.9% of the physicians responded that they would refer their last stage patients to a doctor who is involving palliative care. 8.Only 36.2% of nurses have suggested to physicians that refer the terminally ill patients discharged from the hospital to hospice home care. Once suggested, 95.8% of the physicians have accepted the suggestion. Nurses were reluctant to suggest hospice home care to the physicians, as 48.8% of the nurses said they did not want to. From the result of this study the following conclusion can be drawn, the medical profession's awareness of general hospice care has been increased greatly compared to the results of the previously performed studies. However, this study result also shows that their knowledge of hospice home care is not good enough yet. There is a need for high recommended that medical education institute and develop regular courses on various types of hospice care. Medical field training courses for physicians and nurses will be very helpful as well. It is also important to train hospice experts such as palliative physicians and develop a national hospice management urgently in order to improve the hospice care in Korea.

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Anti-inflammatory effects of Ishige sinicola ethanol extract in LPS-induced RAW 264.7 cell and mouse model (LPS로 유도된 RAW 264.7 Cell과 마우스 모델에 대한 넓패(Ishige sinicola) 에탄올 추출물의 항염증 효과)

  • Kim, Ji-Hye;Kim, Min-Ji;Kim, Koth-Bong-Woo-Ri;Park, Sun-Hee;Cho, Kwang-Su;Kim, Go-Eun;XU, Xiaotong;Lee, Da-Hye;Park, Ga-Ryeong;Ahn, Dong-Hyun
    • Food Science and Preservation
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    • v.24 no.8
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    • pp.1149-1157
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    • 2017
  • Inflammation is the first response of the immune system to infection or irritation in our body. The use of medicinal plants has been widely applied as an alternative source for drug development. One of marine natural resources, the anti-inflammatory effect of Ishige sinicola ethanol extract (ISEE), was evaluated by using LPS-induced RAW 264.7 cell and mice model. As a result, the production of nitric oxide (NO) and pro-inflammatory cytokines (IL-6, IL-$1{\beta}$, TNF-${\alpha}$) were inhibited with increasing concentration of ISEE without any cytotoxicity. Furthermore, ISEE suppressed the expression of not only inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), nuclear factor-kappa B (NF-${\kappa}B$) p65, and mitogen-activated protein kinases (MAPKs), including extracellular signal-regulated kinase (ERK) 1/2, p38, and c-Jun N-terminal kinase (JNK) in a dose-dependent manner. In mice ear edema test, the formation of edema was reduced at the highest dosage of ISEE and the reduction of the number of infiltrated mast cells was observed in histological analysis. These results indicate that ISEE has a potent anti-inflammatory activity and can be used as a pharmaceutical material for many kinds of inflammatory disease.

Apoptotic Cell Death by Pectenotoxin-2 in p53-Deficient Human Hepatocellular Carcinoma Cells (종양억제유전자 p53 결손 인체간암세포에서 Pectenotoxin-2에 의한 Apoptosis 유도)

  • Shin, Dong-Yeok;Kim, Gi-Young;Choi, Byung-Tae;Kang, Ho-Sung;Jung, Jee-H.;Choi, Yung-Hyun
    • Journal of Life Science
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    • v.17 no.10
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    • pp.1447-1451
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    • 2007
  • Through the screening of marine natural compounds that inhibit cancer cell proliferation, we previously reported that pectenotoxin-2 (PTX-2) isolated from marine sponges exhibits selective cytotoxicity against several cell lines in p53-deficient tumor cells compared to those with functional p53. However, the molecular mechanisms of its anti-proliferative action on malignant cell growth are not completely known. To further explore the mechanisms of its anti-cancer activity and to test whether the status of p53 in liver cancer cells correlates with their chemo-sensitivities to PTX-2, we used two well-known hepatocarcinoma cell lines, p53-deficient Hep3B and p53-wild type HepG2. We have demonstrated that PTX-2 markedly inhibits Hep3B cell growth and induces apoptosis whereas HepG2 cells are much more resistant to PTX-2 suggesting that PTX-2 seems to act by p53-independent cytotoxic mechanism. The apoptosis induced by PTX-2 in Hep3B cells was associated with the modulation of DNA fragmentation factor (DFF) family proteins, up-regulation of pro-apoptotic Bcl-2 family members such as Bax and Bcl-xS and activation of caspases (caspase-3, -8 and -9). Blockade of the caspase-3 activity by caspase-3 inhibitor, z-DEVD-fmk, prevented the PTX-2-induced growth inhibition in Hep3B cells. Moreover, treatment with PTX-2 also induced phosphorylation of AKT and extracellular-signal regulating kinase (ERK), but not c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (MARK). Specific inhibitors of PI3K inhibitor (LY294002) and ERK1/2 inhibitor (PD98059) significantly blocks PTX-2-induced-anti-proliferative effects, whereas a JNK inhibitor (SP600125) and a p38 MAPK inhibitor (SB203580) have no significant effects demonstrating that the pro-apoptotic effect of PTX-2 mediated through activation of AKT and ERK signal pathway in Hep3B cells.

Role of PKR and EGR-1 in Induction of Interleukin-S by Type B Trichothecene Mycotoxin Deoxynivalenol in the Human Intestinal Epithelial Cells (B형 트리코테센 곰팡이 독소 데옥시니발레놀에 의한 인체 장관 상피세포 염증성 인터루킨 8유도에서의 PKR과 EGR-1의 상호 역할 규명)

  • Park, Seong-Hwan;Yang, Hyun;Choi, Hye-Jin;Park, Yeong-Min;Ahn, Soon-Cheol;Kim, Kwan-Hoi;Lee, Soo-Hyung;Ahn, Jung-Hoon;Chung, Duk-Hwa;Moon, Yu-Seok
    • Journal of Life Science
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    • v.19 no.7
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    • pp.949-955
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    • 2009
  • Mucosal epithelia sense external stress signals and transmit them to the intracellular cascade responses. Ribotoxic stress-producing chemicals such as deoxynivalenol (DON) or other trichothecene mycotoxins have been linked with gastrointestinal inflammatory diseases by Fusarium-contamination. The purpose of this study was to test the hypothesis that DON evokes the epithelial sentinel signals of RNA-dependent protein kinase (PKR) and early growth response gene 1 (EGR-1), which together contribute to the pro-inflammatory cytokine interleukin 8 (IL-8) in human intestinal epithelial cells. PKR suppression by the dominant negative PKR expression attenuated DON-stimulated interleukin-8 production. Moreover, 1L-8 transcriptional activation by DON was also reduced by PKR inhibition in the human intestinal epithelial cells. Treatment with the PKR inhibitor also suppressed EGR-1 promoter activity, mRNA and protein induction, although mitogen-activated protein (MAP) kinases such as extracellular signal-regulated protein kinases (ERK) 1/2, p38, c-Jun N-terminal Kinase (INK) were little affected or even enhanced in presence of a PKR inhibitor. These patterns were also compared in the EGR-1-suppressed cells, which showed much more suppressed production of 1L-8. All things taken into consideration, DON-activated sentinel signals of EGR-1 via PKR mediated interleukin-8 production in human intestinal epithelial cells, which provide insight into the possible general mechanism associated with mucosal inflammation as an intestinal toxic insult by ribotoxic trichothecene mycotoxins.

Gait Analysis of a Pediatric-Patient with Femoral Nerve Injury : A Case Study (대퇴신경 손상 환아의 보행분석 : 사례연구)

  • Hwang, S.H.;Park, S.W.;Son, J.S.;Park, J.M.;Kwon, S.J.;Choi, I.S.;Kim, Y.H.
    • Journal of Biomedical Engineering Research
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    • v.32 no.2
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    • pp.165-176
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    • 2011
  • The femoral nerve innervates the quadriceps muscles and its dermatome supplies anteromedial thigh and medial foot. Paralysis of the quadriceps muscles due to the injury of the femoral nerve results in disability of the knee joint extension and loss of sensory of the thigh. A child could walk independently even though he had injured his femoral nerve severely due to the penetrating wound in the medial thigh. We measured and analyzed his gait performance in order to find the mechanisms that enabled him to walk independently. The child was eleven-year-old boy and he could not extend his knee voluntarily at all during a month after the injury. His gait analysis was performed five times (GA1~GA5) for sixteen months. His temporal-spatial parameters were not significantly different after the GA2 or GA3 test, and significant asymmetry was not observed except the single support time in GA1 results. The Lower limb joint angles in affected side had large differences in GA1 compared with the normal normative patterns. There were little knee joint flexion and extension motion during the stance phase in GA1 The maximum ankle plantar/dorsi flexion angles and the maximum knee extension angles were different from the normal values in the sound side. Asymmetries of the joint angles were analyzed by using the peak values. Significant asymmetries were found in GA1with seven parameters (ankle: peak planter flexion angle in stance phase, range of motion; ROM, knee: peak flexion angles during both stance and swing phase, ROM, hip: peak extension angle, ROM) while only two parameters (maximum hip extension angle and ROM of hip joint) had significant differences in GA5. The mid-stance valleys were not observed in both right and left sides of vertical ground reaction force (GRF) in the GA1, GA2. The loading response peak was far larger than the terminal stance peak of vertical ground reaction curve in the affected side of the GA3, GA4, GA5. The measured joint moment curves of the GA1, GA2, GA3 had large deviations and all of kinetic results had differences with the normal patterns. EMG signals described an absence of the rectus femoris muscle activity in the GA1 and GA2 (affected side). The EMG signals were detected in the GA3 and GA4 but their patterns were not normal yet, then their normal patterns were detected in the GA5. Through these following gait analysis of a child who had selective injuries on the knee extensor muscles, we could verify the actual functions of the knee extensor muscles during gait, and we also could observe his recovery and asymmetry with quantitative data during his rehabilitation.