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1.
Osteoarthritis Cartilage ; 18(7): 934-41, 2010 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-20346402

RESUMO

OBJECTIVE: The role of postmenopause on the pathogenesis of cartilage degeneration has been an open question. We assessed cartilage degeneration in estrogen receptor (ER)alpha null mice and examined the role of glucocorticoid receptor-interacting protein 1 (GRIP1) in the ERalpha-dependent transcription of a type II collagen gene (col2a1) with special reference to a crosstalk with the transforming growth factor (TGF)-beta signaling pathway. METHODS: The vertebral cartilaginous endplate from female ERalpha null mice was subjected to histological analyses. Col2a1 expression of primary chondrocytes (PCs) obtained from ERalpha null mice after 17beta-estradiol (E(2)) and TGF-beta1 stimulation was examined by reverse transcription polymerase chain reaction (RT-PCR). Estrogen response element (ERE) or col2a1 promoter-enhancer luciferase reporter system was used to investigate the crosstalk among ERalpha, GRIP1, and MKK6. Col2a1 expression and glycosaminoglycan (GAG) content were measured in ATDC5 cells treated with GRIP1 small interfering RNA (siRNA). RESULTS: ERalpha deficiency clearly accelerated impairment of the vertebral cartilaginous endplate. E(2) and TGF-beta1 stimulation increased col2a1 expression in PC from wild-type mice, but not that from ERalpha null mice. The same stimulation increased the col2a1 promoter-enhancer reporter activity, and the elevated activity was decreased by dominant-negative ERalpha and p38 mitogen-activated protein kinase (MAPK) inhibitor. GRIP1 increased the E(2)-dependent ERE activation in the presence of ERalpha and constitutive-active MKK6. GRIP1 siRNA repressed col2a1 expression and GAG production in ATDC5 cells. CONCLUSIONS: Crosstalks between ERalpha/GRIP1 and TGF-beta/MKK6/p38 MAPK pathway have protective roles on cartilage metabolism via regulating the extracellular matrices expression. The finding may lead to the development of a novel therapeutic approach for cartilage degeneration.


Assuntos
Proteínas de Transporte/genética , Cartilagem/metabolismo , Condrócitos/metabolismo , Receptor alfa de Estrogênio/genética , MAP Quinase Quinase 6/genética , Proteínas do Tecido Nervoso/genética , Fatores Etários , Animais , Receptor alfa de Estrogênio/metabolismo , Feminino , Regulação da Expressão Gênica , Disco Intervertebral/metabolismo , MAP Quinase Quinase 6/metabolismo , Camundongos , Modelos Animais , Gravidez , Reação em Cadeia da Polimerase Via Transcriptase Reversa
2.
Minim Invasive Neurosurg ; 53(2): 69-73, 2010 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-20533137

RESUMO

INTRODUCTION: A lumbar discal cyst is a relatively rare cystic lesion that communicates with lumbar intervertebral discs. Surgical resection of the cyst is the reported treatment of choice. In this study, the authors report the minimally invasive surgical resection of lumbar discal cysts using a microendoscopy. PATIENTS AND METHODS: Seven male patients with lumbar discal cysts underwent microendoscopic resections (mean age: 25.1+/-3.2 years and the mean follow-up period: 27.9 months). During the surgeries, the cysts were subtotally resected in a piecemeal fashion, and the fistulas forming the communications between the cysts and the corresponding intervertebral discs were coagulated using a bipolar coagulator. RESULTS: All the patients obtained relief from their pain after surgery, and no recurrences occurred during a mean follow-up period of 28 months. The mean operation time was 72.6+/-20.2 min, and the mean blood loss was 44.4+/-13.7 grams. No intra- or peri-operative complications were noted in any of the patients. CONCLUSIONS: Microendoscopic resection appears to be a minimally invasive and feasible surgical option for the treatment of lumbar discal cysts.


Assuntos
Cistos/cirurgia , Disco Intervertebral/cirurgia , Vértebras Lombares/cirurgia , Doenças da Coluna Vertebral/cirurgia , Adulto , Cistos/patologia , Endoscopia , Humanos , Disco Intervertebral/patologia , Vértebras Lombares/patologia , Masculino , Doenças da Coluna Vertebral/patologia , Resultado do Tratamento
3.
J Clin Invest ; 107(9): 1127-35, 2001 May.
Artigo em Inglês | MEDLINE | ID: mdl-11342576

RESUMO

To study the effects of IL-1 alpha in arthritis, we generated human IL-1 alpha (hIL-1 alpha). Transgenic mice expressed hIL-1 alpha mRNA in various organs, had high serum levels of hIL-1 alpha, and developed a severe polyarthritic phenotype at 4 weeks of age. Not only bone marrow cells but also synoviocytes from knee joints produced biologically active hIL-1 alpha. Synovitis started 2 weeks after birth, and 8-week-old mice showed hyperplasia of the synovial lining layer, the formation of hyperplastic synovium (pannus) and, ultimately, destruction of cartilage. Hyperplasia of the synovial lining was due to the accumulation of macrophage-like cells expressing F4/80 molecules. hIL-1 alpha was widely distributed in macrophage- and fibroblast-like cells of the synovial lining cells, as well as synovial fluid monocytes. T and B cells were rare in the synovial fluid, and analysis of marker expression suggests that synoviocytes were directly histolytic and did not act as antigen-presenting cells. In the joints of these mice, we found elevated levels of cells of the monocyte/macrophage and granulocyte lineages and of polymorphonuclear neutrophils (PMNs), most of which expressed Gr-1, indicating that they were mature, tissue-degrading PMNS: Cultured synoviocytes and PMNs from these animals overexpress GM-CSF, suggesting that the hematopoietic changes induced by IL-1 and the consequent PMN activation and joint destruction are mediated by this cytokine.


Assuntos
Artrite/etiologia , Interleucina-1/biossíntese , Macrófagos , Neutrófilos , Membrana Sinovial/patologia , Animais , Artrite/genética , Artrite/imunologia , Artrite/patologia , Linhagem da Célula , Fator Estimulador de Colônias de Granulócitos e Macrófagos , Células-Tronco Hematopoéticas , Humanos , Interleucina-1/genética , Camundongos , Camundongos Transgênicos , Monócitos , Fenótipo , Proteínas Recombinantes/biossíntese
4.
Biochim Biophys Acta ; 1350(3): 253-8, 1997 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-9061018

RESUMO

Expression patterns for the two isoforms of alpha 1(II) mRNA in various cartilaginous tissues were examined using newly isolated cDNA clones encoding rabbit type II procollagen amino- and carboxy-terminal propeptide regions. In nonchondrogenic nucleus pulposus, the switching of the mRNA from the long form to the short form was accompanied by disc maturation after birth. Interestingly, the short transcript was also expressed preferentially in human chordoma tissues as aberrant chordal vestiges. These results suggest an abundance of the differentiated chondrocyte-like phenotype in the heterogeneous notochordal remnants.


Assuntos
Processamento Alternativo , Cartilagem/química , Regulação da Expressão Gênica no Desenvolvimento , Notocorda/química , Pró-Colágeno/genética , RNA Mensageiro/análise , Idoso , Animais , Animais Recém-Nascidos , Sequência de Bases , Cartilagem/embriologia , Cordoma/química , DNA Complementar/genética , Humanos , Mesoderma/química , Dados de Sequência Molecular , Notocorda/embriologia , Especificidade de Órgãos , RNA Mensageiro/genética , Coelhos , Homologia de Sequência do Ácido Nucleico
5.
Eur J Cancer ; 36(4): 496-502, 2000 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-10717526

RESUMO

The aim of the study was to clarify the role of telomerase component genes in hepatocarcinogenesis and to examine both the relationship between the expression of telomerase component genes and histological differentiation in hepatocellular carcinoma (HCC) and the relationship between expression levels of telomerase component genes and telomerase activity in HCCs. Telomerase is a ribonucleoprotein enzyme composed of a template RNA and several proteins. Recently, three such telomerase component genes have been identified: human telomerase reverse transcriptase (hTERT); human telomerase RNA component (hTERC); and telomerase-associated protein 1 (TEP1). The expression of these components was evaluated in 34 HCCs and 24 non-cancerous liver tissues by reverse transcriptase-polymerase chain reaction (RT-PCR). Expression of hTERT mRNA was detected in most HCCs, but not in the non-cancerous tissues (P<0.01). Expression of hTERC was detected in both HCCs and non-cancerous tissues, but the expression level in HCCs was higher than that in non-cancerous tissues (P<0.01) and tended to increase as histological differentiation became less marked. The expression level of hTERT mRNA correlated with relative telomerase activity (P<0.01). These results suggest that telomerase reactivation during hepatocarcinogenesis might be regulated by only hTERT and an increase in telomerase activity level in tumour progression might be regulated by both hTERT and hTERC.


Assuntos
Carcinoma Hepatocelular/enzimologia , Neoplasias Hepáticas/enzimologia , Telomerase/genética , Adulto , Idoso , Carcinoma Hepatocelular/genética , Feminino , Regulação Enzimológica da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Hepatite/metabolismo , Humanos , Fígado/metabolismo , Cirrose Hepática/metabolismo , Neoplasias Hepáticas/genética , Masculino , Pessoa de Meia-Idade , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Telomerase/metabolismo
6.
J Orthop Res ; 15(4): 528-38, 1997 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-9379262

RESUMO

To clarify phenotypic alterations of intervertebral disc cells during the repair process, we cloned partial type-II collagen cDNA from rabbits and analyzed the level of expression of type-II collagen mRNA in disc degeneration. An animal model was created by surgical denucleation of rabbit intervertebral discs through an extraperitoneal approach. Eight animals each from an experimental and a control group were killed at 2, 4, 8, or 16 weeks postoperatively, and the disc samples were used for this study. Round chondrocyte-like cells that filled the herniated space showed intense signal of type-II collagen mRNA and significant pericellular immunostaining of type-II collagen but no clear staining of type-I collagen. Northern blot analysis revealed that the expression of type-II collagen mRNA of the repair disc cells was transiently increased at 4 weeks postoperatively. The cells were able to change their morphology in response to mechanical stimulation by surgical denucleation and to induce a significant increase in the gene expression of type-II collagen at an early phase of disc degeneration. The present results indicate the transient enhancement of repair activity in the degenerative process of injured fibrocartilage.


Assuntos
Colágeno/genética , Disco Intervertebral/patologia , Animais , Northern Blotting , Cartilagem Articular/química , Cartilagem Articular/patologia , Condrócitos/química , Condrócitos/fisiologia , Clonagem Molecular , Colágeno/análise , DNA Complementar/isolamento & purificação , Feminino , Expressão Gênica/fisiologia , Hibridização In Situ , Disco Intervertebral/citologia , Disco Intervertebral/cirurgia , Fenótipo , Pró-Colágeno/análise , Pró-Colágeno/genética , RNA Mensageiro/análise , Coelhos , Regulação para Cima
7.
J Gastroenterol ; 30 Suppl 8: 121-3, 1995 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-8563872

RESUMO

Low doses of 6-mercaptopurine (6MP) were used for the treatment of inflammatory bowel disease, and 20-30 mg/day was found to be effective for patients with ulcerative colitis who were corticosteroid-dependent or corticosteroid-resistant. Corticosteroid was tapered in 20 of 21 patients with ulcerative colitis. Of 15 patients who were refractory to conventional therapy, 11 responded to 6MP treatment. The same doses of 6MP were given to patients with Crohn's disease who were corticosteroid-dependent or who had associated fistula. Treatment with 6MP did not influence the changes in colonic or ileac lesions in Crohn's disease. However, the fistulas were closed or improved in 70% of 10 patients by 6MP treatment. The adverse effects of small doses of 6MP were minimal. These results confirm that immunosuppressive agents are effective for patients with inflammatory bowel disease. In a rat colitis model induced by immunization with trinitrobenzene (TNB), we used anti-CD4 monoclonal antibodies to prevent colonic inflammation; these antibodies were effective for this colitis model, suggesting that a novel therapy targeting CD4 intestinal lymphocytes may be feasible in the treatment of Crohn's disease.


Assuntos
Colite Ulcerativa/tratamento farmacológico , Doença de Crohn/tratamento farmacológico , Imunossupressores/uso terapêutico , Mercaptopurina/uso terapêutico , Animais , Anticorpos Monoclonais/uso terapêutico , Antígenos CD4/imunologia , Colite/induzido quimicamente , Colite/terapia , Humanos , Ratos , Indução de Remissão , Ácido Trinitrobenzenossulfônico
8.
J Gastroenterol ; 30 Suppl 8: 73-5, 1995 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-8563896

RESUMO

To analyze the nature of intestinal mucosal lymphocytes in Crohn's disease, we established T cell lines of patients' intraepithelial lymphocytes. T cell lines from the affected terminal ileum of the patients showed an increased proportion of CD4+V beta 5.2/5.3+ T cells. These cells were increased in number after stimulation with staphylococcal enterotoxins C1 and D, showed an increase in cytolytic activity, and produced a large amount of interferon-gamma. To clarify the role of CD4+ mucosal lymphocytes in the intestinal inflammation, we then developed a novel colitis model by immunizing a rat with trinitrobenzenesulfonic acid (TNB) emulsion with adjuvant. Deep ulceration and granuloma formation in this colitis model resembled the histopathological findings of human Crohn's disease. Immunohistochemical and flow cytometric analysis demonstrated that the number of CD45RC(high)CD4+ mucosal lymphocytes was increased. Interestingly, the administration of anti-CD4 Abs prevented severe inflammation in the model. After treatment with anti-CD4 Abs, the anti-TNB Ab titer, the number of CD45RC(high)CD4+ cells, and interferon-gamma mRNA expression were significantly decreased in the mucosa of the model. These results suggest that some subsets of CD4+ mucosal lymphocytes play an important role in the triggering and progression of inflammation in Crohn's disease.


Assuntos
Linfócitos T CD4-Positivos/imunologia , Doença de Crohn/etiologia , Animais , Células Cultivadas , Colite/imunologia , Colo/imunologia , Colo/patologia , Doença de Crohn/imunologia , Doença de Crohn/patologia , Feminino , Humanos , Íleo/imunologia , Íleo/patologia , Mucosa Intestinal/imunologia , Mucosa Intestinal/patologia , Ratos , Ratos Wistar
9.
J Gastroenterol ; 35(1): 20-7, 2000.
Artigo em Inglês | MEDLINE | ID: mdl-10632536

RESUMO

To clarify the role of colonic mucin in the autoimmune process of ulcerative colitis, circulating antibodies against human colonic mucin were investigated. Purified colonic mucin, obtained from human colonic mucosa by gel filtration, using a Bio-Gel A-1.5-m column and CsCl equilibrium density gradient, was divided into soluble mucin (S-mucin) secreted extracellularly and membranous mucin (M-mucin) binding to cell membrane. Sodium dodecylsulfate polyacrylamide gel electrophoresis and Western blotting analysis showed that antibodies in the serum samples of some patients with ulcerative colitis recognized purified S- and M-mucin of >180-kD. By enzyme-linked immunosorbent assay (ELISA), anti-mucin antibodies were detected in 11 of 60 patients with ulcerative colitis (18%). In contrast, the antibodies were not detected in 22 patients with Crohn's disease. The titers of antimucin antibodies against S-mucin and M-mucin were not different in each patient. By ELISA using mucin in which the sugar chains were destroyed by neuraminidase or NaIO4 treatment, it was demonstrated that anti-mucin antibodies recognized the epitopes of either the sugar chain or the core protein exposed through destruction of the sugar chains. We then investigated the relationship between anti-mucin antibodies and the patients' clinical features. Anti-mucin antibodies were detected in 6 of 15 patients with chronic continuous type ulcerative colitis (40%) and in 5 of 26 patients with relapsing-remitting type (19%), but there was no antimucin antibody-positive serum in patients who had had only one attack without any relapse. These results suggest that anti-mucin antibodies could be a disease marker for ulcerative colitis and that immunological abnormalities in colonic mucin contribute to the persistence of colonic mucosal inflammation.


Assuntos
Autoanticorpos/imunologia , Colite Ulcerativa/imunologia , Mucinas/imunologia , Western Blotting , Colo/imunologia , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Masculino
10.
Nihon Rinsho ; 56(5): 1239-43, 1998 May.
Artigo em Japonês | MEDLINE | ID: mdl-9613130

RESUMO

Telomerase activity and terminal restriction fragment (TRF) length were examined in hepatocellular carcinoma (HCC). Telomerase activity was assayed by telomeric repeat amplification protocol (TRAP) connected with an internal telomerase assay standard (ITAS). The incidence of strong telomerase activity (highly variable level compared with the activity of non-cancerous liver tissue) was 79% in well, 84% in moderately, and 100% in poorly differentiated HCC, while 0% in non-cancerous liver tissues. The incidence of TRF length alteration (reduction or elongation) was 53% in HCC. The incidence of TRF alteration was significantly higher in HCC exceeding 3 cm in diameter, moderately or poorly differentiated in histology. Telomerase activity was not associated with TRF length alteration in HCC. In conclusion, strong telomerase activity and TRF length alteration increased with HCC tumor progressions.


Assuntos
Carcinoma Hepatocelular/genética , Neoplasias Hepáticas/genética , Telomerase/análise , Telômero/genética , Carcinoma Hepatocelular/patologia , Diferenciação Celular , Humanos , Neoplasias Hepáticas/patologia
18.
Spinal Cord ; 46(4): 282-6, 2008 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-17909556

RESUMO

STUDY DESIGN: Retrospective case series. OBJECTIVE: To evaluate our recent treatment strategy for intramedullary spinal cord tumors. SETTING: Department of Orthopaedic Surgery, Keio University, Japan. METHODS: We reviewed 68 cases of intramedullary tumors (ependymoma, 33; astrocytoma, 23; hemangioblastoma, 12), treated surgically between 1994 and 2003. There were 42 males and 26 females whose mean age at the time of surgery was 43 years. The mean follow-up period was 6.2 years. The tumor malignancy grade according to the WHO classification was astrocytoma grade I, 3; grade II, 8 (low-grade: 11 cases); grade III, 10; grade IV, 2 (high-grade: 12 cases). All ependymomas were grade II. Three of the 12 hemangioblastomas were associated with von Hippel-Lindau disease. RESULTS: Total excision was achieved in 90% of the ependymomas and functional improvement was obtained when the preoperative neurological deficit was mild. Approximately 50% of low-grade astrocytomas could be totally excised with favorable survival outcomes, suggesting that total excision should be attempted for low-grade astrocytomas. However, total excision of high-grade tumors was difficult and the functional outcomes were poor. Cordotomy should be considered in patients with a thoracic high-grade astrocytoma. Total resection was possible in 92% of hemangioblastoma, and the functional outcomes were good, however, more attention should be paid for tumors with feeding arteries on the ventral side and for those associated with von Hippel-Lindau disease. CONCLUSIONS: Predictors of good surgical outcome for intramedullary spinal cord tumors were histological grades of the tumors, surgical margins, and neurological status of the patient before surgery.


Assuntos
Astrocitoma/cirurgia , Ependimoma/cirurgia , Hemangioblastoma/cirurgia , Neoplasias da Medula Espinal/cirurgia , Adolescente , Adulto , Idoso , Astrocitoma/mortalidade , Astrocitoma/patologia , Vértebras Cervicais , Criança , Pré-Escolar , Estudos de Coortes , Ependimoma/mortalidade , Ependimoma/patologia , Feminino , Hemangioblastoma/mortalidade , Hemangioblastoma/patologia , Humanos , Masculino , Pessoa de Meia-Idade , Complicações Pós-Operatórias , Estudos Retrospectivos , Neoplasias da Medula Espinal/mortalidade , Neoplasias da Medula Espinal/patologia , Taxa de Sobrevida , Vértebras Torácicas , Resultado do Tratamento
19.
Minim Invasive Neurosurg ; 50(6): 335-9, 2007 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-18210355

RESUMO

The purpose of this study was to elucidate the feasibility of microendoscopic discectomy (MED) for the treatment of lumbar disc herniation with a bony fragment due to apophyseal separation. Eighteen patients with low back pain and unilateral sciatic pain due to lumbar disc herniation with a bony fragment were treated by MED using the unilateral approach (15 males and three females; mean age, of 28.9 years; mean follow-up period, 21.1 months); 18 age-and sex-matched patients with lumbar disc herniation without a bony fragment treated by MED served as the control group. The clinical outcomes were evaluated using the Japanese Orthopedic Association Score for Low Back Pain (JOA scores; maximum score, 29 points). Evaluation of the results revealed that good surgical outcomes equivalent to those in the control group were obtained in the subjects of LDH with a bony fragment (JOA scores; 14.1+/-3.5 in the patient group vs.15.4+/-2.6 in the control group before surgery; 26.3+/-1.8 in the patient group vs. 26.9+/-1.3 at follow-up after the surgery). Although the mean surgical time was significantly longer in the patient group, there were no intra- or postoperative complications in either group. We conclude that MED using the unilateral approach is a feasible minimally invasive surgical option for patients of lumbar disc herniation with an apophyseal bony fragment.


Assuntos
Endoscopia/métodos , Deslocamento do Disco Intervertebral/cirurgia , Disco Intervertebral/cirurgia , Vértebras Lombares/cirurgia , Microcirurgia/métodos , Fraturas da Coluna Vertebral/cirurgia , Adolescente , Adulto , Idoso , Criança , Endoscopia/estatística & dados numéricos , Estudos de Viabilidade , Feminino , Humanos , Disco Intervertebral/patologia , Deslocamento do Disco Intervertebral/complicações , Deslocamento do Disco Intervertebral/patologia , Vértebras Lombares/patologia , Imageamento por Ressonância Magnética , Masculino , Microcirurgia/estatística & dados numéricos , Pessoa de Meia-Idade , Procedimentos Neurocirúrgicos/métodos , Procedimentos Neurocirúrgicos/estatística & dados numéricos , Complicações Pós-Operatórias/etiologia , Fraturas da Coluna Vertebral/complicações , Fraturas da Coluna Vertebral/patologia , Resultado do Tratamento
20.
Spinal Cord ; 44(12): 740-5, 2006 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-16670687

RESUMO

STUDY DESIGN: Retrospective case series. OBJECTIVES: To analyze prognostic factors for patients with spinal cord astrocytomas. SETTING: Department of Orthopaedic Surgery, Keio University, Japan. METHODS: Seven patients received total excisions (group T), eight underwent partial excisions (group P), and 15 had excisional biopsies (group B). Impacts of the tumor histological grade, the level of the tumor, the types of surgical interventions, and the use of adjuvant radiotherapies on the survival and functional outcomes of 30 patients (18 in low-grade and 12 high-grade malignancy tumors) were analyzed. RESULTS: The survival rate of the low-grade malignancy group was significantly higher than that of the high-grade group. The survival rate of the patients with thoracic astrocytomas was significantly higher than those with cervical astrocytomas. In both the low- and high-grade groups, the survival rates in groups P/T were significantly higher than those in group B. In the low-grade group, five patients, whose preoperative functional statuses were classified as 'fair' or better, remained 'fair' or better after surgery. In the high-grade group, the postoperative functional statuses were classified as 'no change' or 'aggravated' in all except two patients. No significant difference in the survival rates was detected between patients with and without adjuvant radiotherapy. CONCLUSIONS: The tumor grade and the extent of tumor resection were significant prognostic factors for survival rate. In low-grade malignancy group, good motor function was retained when surgeries were performed before substantial neurological deterioration. The efficacy of postoperative radiotherapy has yet to be determined and needs further study.


Assuntos
Astrocitoma/cirurgia , Neoplasias da Medula Espinal/cirurgia , Adolescente , Adulto , Astrocitoma/patologia , Criança , Feminino , Humanos , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Prognóstico , Neoplasias da Medula Espinal/patologia , Estatísticas não Paramétricas , Taxa de Sobrevida , Resultado do Tratamento
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