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The purpose of the study was to translate the athlete burnout questionnaire (ABQ) into Simplified Chinese and examine its psychometric properties in Chinese collegiate athletes and elite athletes. Firstly, the factor structure, internal consistency reliability and nomological validity of the Chinese translated ABQ was examined in a sample of Chinese collegiate athletes (n = 214, 58.9% females). Secondly, abovementioned psychometric properties were examined in a sample of Chinese elite athletes (n = 505, 52.7% females). Finally, measurement invariance of the Chinese translated ABQ was examined across the two samples. It was found that the 12-item three-correlated-factors model outperformed the one factor model and bi-factor model in collegiate athlete sample whereas the 12-item bi-factor model best represented the factor structure of the Chinese translated ABQ in elite athlete sample. Satisfactory internal consistency reliabilities of the Chinese translated ABQ were evidenced in the two samples. Nomological validity was also supported by the results of the two samples that the three subscales of the ABQ were significantly associated with its theoretically related variables. Results of multiple-group confirmatory factor analysis revealed that weak measurement invariance of the Chinese translated ABQ (three-correlated-factors model) was evidenced across the two samples. Collectively, results of this study indicated that the 12-item Chinese translated ABQ could be used for measuring burnout of Chinese collegiate and elite athletes. Significance and implication of the current study as well as recommendations for future study were discussed.
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OBJECTIVE: To observe the effect of warming needling therapy on gastrointestinal reaction after hyperthermic intraperitoneal chemotherapy (HIPEC) in patients of spleen and stomach deficiency syndrome after colon cancer surgery. METHODS: A total of 120 cases of HIPEC were randomized into observation group and control group, 60 cases in each. The patients of the two groups all received HIPEC. In the observation group, 1 h before HIPEC, warming needling technique was applied to Zusanli (ST36), Sanyinjiao (SP6) and Yinlingquan (SP9) and the even-needing technique of acupuncture was applied to Neiguan (PC6) for 30 min, and then the intravenous injection with Ondansetron was given 30 min before HIPEC. In the control group, The intravenous injection with Ondansetron was given 30 min before HIPEC. In the two groups, the changes in nausea, vomiting, abdominal distention, diarrhea, total bilirubin (TB), alanine aminotransferase (ALT), alkaline phosphatase (ALP) and KPS score, as well as the average hospitalization length of stay were observed. RESULTS: The total effective rates in the treatment of nausea, vomiting, abdominal distention and diarrhea were 91.67%(55/60), 93.33%(56/60), 80.00%(48/60) and 88.33%(53/60) in the observation group and were 78.33%(47/60), 78.33%(47/60), 63.33%(38/60) and 70.00%(42/60) in the control group respectively, and the total effective rate in the treatment of gastrointestinal reaction of HIPEC in the observation group was obviously higher than that in the control group(P<0.05). The KPS score and curative effect in the observation group was obviously higher than those of the control group(P<0.05), and the average hospitalization length of stay in the observation group was obviously reduced as compared with the control group (P<0.05). There were no significant differences in serum TB, ALT and ALP contents between the two groups after treatment (P>0.05). CONCLUSION: The warming needling technique of acupuncture and moxibustion alleviates the gastrointestinal reaction, improves KPS score and reduces the hospitalization length of stay in HIPEC patients after the surgery of colon cancer and differentiated as the spleen and stomach deficiency in traditional Chinese medicine.
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Terapia por Acupuntura , Neoplasias do Colo , Pontos de Acupuntura , Neoplasias do Colo/terapia , Humanos , Náusea , Resultado do TratamentoRESUMO
Relationship between methylenetetrahydrofolate reductase (MTHFR) A1298C gene polymorphism and type 2 diabetic nephropathy (T2DN) risk is still unclear. This study was performed to evaluate if there is an association between the MTHFR A1298C gene polymorphism and T2DN risk using meta-analysis. The relevant reports were searched and identified from PubMed, Cochrane Library on 1 October 2013, and eligible studies were included and synthesized. Eight reports were recruited into this meta-analysis for the association of the MTHFR A1298C gene polymorphism with T2DN risk. The MTHFR A1298C C allele or CC genotype was shown to be not associated with T2DN risk (C allele: OR = 0.76, 95% CI: 0.43-1.34, p = 0.34; CC genotype: OR = 1.18, 95% CI: 0.63-2.22, p = 0.60). Interestingly, AA genotype was associated with the T2DN risk (OR = 0.68, 95% CI: 0.49-0.96, p = 0.03). In the sensitivity analysis according to the Hardy-Weinberg equilibrium (HWE), the results were consistent with those in non-sensitivity analysis. However, in the sensitivity analysis according to the control source from hospital, sample size of case (≥ 100), sample size of case (<100), the MTHFR A1298C gene polymorphism was not associated with T2DN risk. In conclusion, the MTHFR A1298C gene polymorphism was not associated with T2DN risk. However, additional studies are required to firmly establish a correlation between the MTHFR A1298C gene polymorphism and T2DN risk.
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Diabetes Mellitus Tipo 2/complicações , Nefropatias Diabéticas/genética , Metilenotetra-Hidrofolato Redutase (NADPH2)/genética , Predisposição Genética para Doença , Humanos , Polimorfismo de Nucleotídeo ÚnicoRESUMO
OBJECTIVE: To investigate the efficacy and advantages of laparoscopy-assisted total gastrectomy (LATG) with D2 dissection of lymph nodes versus conventional open D2 total gastrectomy (OTG) in advanced gastric cancer. METHODS: One hundred and twenty-five patients with advanced gastric cancer in the middle or upper third of the stomach were operated on from July 2005 to March 2007. Of the patients, 59 cases received LATG and 66 OTG with D2 lymph nodes dissection. Clinical data were recorded and compared between the two groups. RESULTS: No patient in the LATG group converted to conventional operation with laparotomy. No operation mortality and no severe morbidity occurred in LATG group. As compared with OTG group, in LATG group operation time was longer [(330 +/- 71) min vs. (261 +/- 54) min, P =0.005] in LATG group, but with similar number of lymph node retrieval (36 +/- 13 vs. 34 +/- 16, P =0.450), less operation blood loss [(175 +/- 101) ml vs. (359 +/- 210) ml, P =0.003], earlier recovery of bowel activity (P = 0.015), and a shorter duration of fever after operation (P = 0.024). CONCLUSIONS: LATG with D2 lymph node dissection in advanced gastric cancer is safe and technically feasible with better operative access and visual field, less operation blood loss and earlier recovery.
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Gastrectomia/métodos , Laparoscopia , Neoplasias Gástricas/cirurgia , Adulto , Idoso , Estudos de Viabilidade , Feminino , Humanos , Excisão de Linfonodo , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Neoplasias Gástricas/patologia , Resultado do TratamentoRESUMO
OBJECTIVE: To investigate the management of chylous leakage after radical operation of gastric carcinoma. METHODS: 161 patients with gastric carcinoma underwent D2-D4 dissection. A double catheterization cannula was employed in each patient around the abdominal aorta above the celiac trunk and crus of diaphragm. Postoperatively, the chylous fluid from the drainage tube was observed, smeared and cultured; infection of chylous fluid was treated. The development of chylous leakage was observed and the optimal time to remove the drainage tube was determined. RESULTS: Chylous leakage occurred in 19 patients. The volume of chylous leakage was less than 250 ml/24 h in 8 patients, 250 - 500 ml/24 h in 7, and 500 - 1500 ml/24 h in 4. Candida albicans was founded in the fluid of chylous leakage in 8 patients, and bacterial infection was found simultaneously in 5 of them. The patients with chylous leakage were healed within 10 - 90 postoperative days. The drainage tube was removed when there was no fluid in the tube and no hydrops in peritoneal cavity by B ultrasound, and the patient were in good condition without signs and symptoms of infections. CONCLUSION: Chylous leakage after D2 - D4 dissection for gastric carcinoma can be cured by immediate diagnosis, thorough drainage, and anti-infectious treatment with regional and continuative washout when the chylous fluid is infected by Candida or bacteria.
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Ascite Quilosa/terapia , Gastrectomia/efeitos adversos , Complicações Pós-Operatórias/terapia , Neoplasias Gástricas/cirurgia , Adulto , Idoso , Anti-Infecciosos/uso terapêutico , Bactérias/efeitos dos fármacos , Candida/efeitos dos fármacos , Ascite Quilosa/diagnóstico , Ascite Quilosa/etiologia , Drenagem , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Cuidados Pós-Operatórios/métodos , Complicações Pós-Operatórias/diagnóstico , Resultado do TratamentoRESUMO
OBJECTIVE: To clone novel gene from suppression subtraction library established for screening down-regulated genes in gastric carcinoma, and the effects of novel gene on gastric tumorigenicity were analyzed. METHODS: Sequencing results of 860 positive colonies chosen randomly were compared by Blast program in GenBank. Novel gene fragment was amplified by rapid amplification of cDNA ends (RACE). The mRNA expression of novel gene was detected by Northern blot and semi-quantitative PCR in 25 cases of gastric carcinoma tissue and counterpart normal gastric mucosa. The structure and chromosomal location of novel gene were investigated by Bio-message technique. RESULTS: A 233 bp novel gene fragment was screened out from 860 clones and a 802 bp novel gene was obtained by RACE. The novel gene was named as GDDM, registered in the number of AF494508 by GenBank. The mRNA expression of GDDM in gastric carcinoma tissue (4.496+/-0.637) was significantly lower than that in the counterpart normal gastric mucosa (36.919+/-6.290)(P<0.01). Chromosomal location of GDDM gene was at 4q31. CONCLUSION: The cloned novel gene, GDDM, is down-regulated in gastric carcinoma, and it is likely to be involved in gastric tumorigenicity.
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Clonagem Molecular , Genes Neoplásicos , Neoplasias Gástricas/genética , Sequência de Bases , DNA Complementar , Regulação para Baixo , Amplificação de Genes , Regulação Neoplásica da Expressão Gênica , Biblioteca Gênica , Humanos , Dados de Sequência Molecular , Neoplasias Gástricas/metabolismoRESUMO
Lycopene liposomes were prepared by conventional rotary-evaporated film-ultrasonication method. The release of lycopene from lycopene liposome was evaluated in vitro. The pharmacokinetic parameters of lycopene liposomes (L-LYC) and lycopene (LYC) oil, the effect of LYC and L-LYC on antioxidation were also investigated in rats. HPLC method was used to assay the concentration of lycopene in rat's plasma. Pharmacokinetic parameters were estimated by 3P97 program. The release of L-LYC and LYC were measured in the artificial stomach liquid and bowel liquid. After 4 weeks of L-LYC or LYC feeding, the activity of SOD, T-AOC, GSH-Px, MDA and CAT in serum and liver were measured separately. The pharmacokinetic parameters of LYC oil and L-LYC in a single dose were 4.45 and 7.45 h for Tmax; 0.473 and 0.654 microg x mL(-1) for Cmax; 12.38 and 21.67 mirog x h x mL(-1) for AUC,respectively. The activities of GSH-Px and T-AOC in serum and liver of the L-LYC group increased (P < 0.05) and the concentrations of MDA and CAT decreased significantly (P < 0.05). It could be concluded that lycopene liposomes could prolong the time of absorption. L-LYC could increase antioxidative effect and reduce lipid peroxidation obviously compared with LYC in rats.
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Antioxidantes/farmacocinética , Carotenoides/farmacocinética , Portadores de Fármacos , Lipossomos/química , Administração Oral , Animais , Antioxidantes/administração & dosagem , Área Sob a Curva , Carotenoides/administração & dosagem , Catalase/sangue , Catalase/metabolismo , Cromatografia Líquida de Alta Pressão/métodos , Glutationa Peroxidase/sangue , Glutationa Peroxidase/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Fígado/metabolismo , Licopeno , Masculino , Malondialdeído/sangue , Malondialdeído/metabolismo , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Superóxido Dismutase/sangue , Superóxido Dismutase/metabolismoRESUMO
OBJECTIVE: To confirm the GDDR cDNA property of novel down-regulated full-length gene in gastric cancer, structure of genomic GDDR DNA and its promotor region. To predict its transcription factors and transcription factor binding sites. To explore function of GDDR gene in vitro. METHODS: GDDR mRNA was located by in situ mRNA hybridization of gastric mucous membranes, and was amplified in 13 human organs and tissues. The structure and location of GDDR on chromosome, property of protein encoded by full-length GDDR were investigated by Bio-message technique. Promotor region of GDDR was confirmed, and transcription factors or their binding sites were predicted in software Gene2promoter and Matinspector of Genomatix. Both of vector pcDNA3.1/Myc-His(-)A inserted by GDDR ORF and control vector pcDNA3.1/Myc-His(-)A were respectively transfected into gastric cell lines 7901 by lipofectamin. Growth curve, MTT test and a morphological analysis were respectively performed. RESULTS: GDDR mRNA was located in gastric mucous epithelial cells, and only was expressed in gastric tissue. 7739 bp genomic GDDR DNA located on chromosome 2p13.3, 21701 bp away from CA11-one stomach-specific gene related to gastric cancer. 618 bp promotor region of GDDR located at position +96 bp,and -419 bp of transcription start site of GDDR. The structure of genomic DNA or cDNA between gene GDDR and CA11 was mostly similar. Sequences of their promotor region were different, transcription factors and their binding sites also varied between gene GDDR and CA11. GDDR encoded protein including a trans-membrane peptide homologed to CA11 that have been proven to encode secrete protein. GDDR was another new member of BRCHOS family just was found. Gastric cell lines 7901 transfected by GDDR showed a marked decrease in growth rate by growth curve and MTT test (72 h, 0.341 +/- 0.014 vs 0.488 +/- 0.015 A, P < 0.01). CONCLUSIONS: Stamoch-specific, novel down-regulated gene GDDR in gastric cancer locates in gastric mucous epithelial cells can markedly inhibit growth of gastric cancer cell lines 7901, GDDR is another new member of BRICHOS family related to gastric cancer except CA11.
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Proteínas de Membrana/genética , Neoplasias Gástricas/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Proteínas de Transporte , Cromossomos Humanos Par 2/genética , DNA Complementar/química , DNA Complementar/genética , Regulação para Baixo , Humanos , Técnicas In Vitro , Proteínas de Neoplasias , Regiões Promotoras Genéticas/genética , Células Tumorais CultivadasRESUMO
Following contents were reviewed in this article: More and more experimental studies related with chronic uratic nephropathy were carried out in recent years. In most of these studies, the animal models were established mainly from viewpoints of gene recombination, urinary uric acid inhibition and blood uric acid production promoting. TCM showed good effects in lowering blood uric acid, regulating levels of cytokines and postponing interstitial fibrosis. However, further studies on Chinese herbs and their extracts is necessary.
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Modelos Animais de Doenças , Medicamentos de Ervas Chinesas/farmacologia , Nefropatias , Animais , Medicamentos de Ervas Chinesas/uso terapêutico , Hiperuricemia/sangue , Nefropatias/etiologia , Camundongos , Fitoterapia , Purinas/metabolismo , Ratos , Ácido Úrico/sangueRESUMO
AIM: To investigate the expression of human telomerase reverse transcriptase(hTERT) gene and its relationship with proliferating cell nuclear antigen and P53. METHODS: Immunohistochemical staining was used to detect the expressions of hTERT, PCNA and P53 in 42 hepatocellular carcinoma(HCC) tissues. The relationship between hTERT expression, pathological characters of HCC and PCNA and P53 expression was analyzed. RESULTS: The expression rates of hTERT, PCNA and P53 in HCC tissues were 71.4%(30/42),76.2%(32/42) and 73.8%(31/42), respectively. hTERT was not expressed in normal liver tissues. The expression rates of hTERT gene in HCC tissues of different pathological grades (I, II and III)were 40.0%(4/10), 70.0%(14/20) and 100%(12/12), respectively. The expression of hTERT was correlated with HCC recurrence. CONCLUSION: The expression of hTERT gene may relate to the genesis and progression of HCC. There is no significant correlation between the expression of hTERT and PCNA and P53. The detection of hTERT gene expression may be regarded as a marker for recurrence of HCC.
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Carcinoma Hepatocelular/metabolismo , Proteínas de Ligação a DNA/metabolismo , Neoplasias Hepáticas/metabolismo , Antígeno Nuclear de Célula em Proliferação/metabolismo , Telomerase/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Idoso , Biomarcadores Tumorais , Carcinoma Hepatocelular/patologia , Feminino , Humanos , Fígado/metabolismo , Neoplasias Hepáticas/patologia , Masculino , Pessoa de Meia-Idade , Recidiva Local de Neoplasia/metabolismoRESUMO
OBJECTIVE: To set up a stand for surgical classification of pancreatic duct stone and evaluate the benefits of different management according to the classification. METHODS: Retrospectively analysis the diagnosis and prognosis of different management of 33 cases pancreatic duct stones to establish a new standard of classification and strategy of management of pancreatic duct stone. RESULTS: According to the results of imaging examination (B-US, CT, ERCP) and finding during surgery, pancreatic duct stone can be classified into four different types: Type I: The stones mainly located in the head of pancreas. Endoscopic pancreas drainage and remove of stones is the first line choice of treatment. If it fail the Whipple procedure should be applied. Type II, The stones mainly located in the body of pancreas. It can be treated by Pusetow procedure. Type III, The stones mainly located in the tail of pancreas. The resection of the tail of pancreas or combined with spleenectomy was recommended for the management of this type stones. Type IV, The stones can be found from the head to tail of the main duct of pancreas. The Pusetow-Gillesby procedure or dividing of the neck of pancreas removing stones from both ends of pancreatic duct and reconstructed by two ends pancreatic duct-ileostomy in Roux-en-Y fashion are the choice of management. CONCLUSION: The invadulaized strategy of the management based upon correct diagnosis and classification play the most important role in the treatment of pancreatic duct stone.
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Cálculos/classificação , Cálculos/cirurgia , Ductos Pancreáticos/patologia , Adulto , Idoso , Cálculos/diagnóstico , Colangiopancreatografia Retrógrada Endoscópica , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Pancreatopatias/classificação , Pancreatopatias/diagnóstico , Pancreatopatias/cirurgia , Ductos Pancreáticos/diagnóstico por imagem , Estudos Retrospectivos , Tomografia Computadorizada por Raios X , UltrassonografiaRESUMO
AIM: To investigate the hepatocellular apoptosis after hepatectomy in obstructive jaundice and biliary decompression rats. METHODS: After bile duct ligation for 7 days, rats were randomly divided into OB group in which the rats underwent 70% hepatectomy, OB-CD group in which the rats underwent hepatectomy accompanied by choledochoduodenostomy, CD-Hx group in which the rats underwent choledochoduodenostomy and then received 70% hepatectomy on the fifth day after biliary decompression. The control group (Hx group) only underwent hepatectomy. RESULTS: The level of total serum bilirubin and serum enzymes was significantly lower in CD-Hx group than in OB-CD and OB groups on day 1, 3 and 5 after hepatectomy. The apoptotic index was significantly lower in CD-Hx group than in OB-CD and OB groups on day 3 and 5. The oligonucleosomal DNA fragments and Caspase-3 activity were also lower in CD-Hx group than in OB-CD and OB groups 3 days after hepatectomy, without differences between CD-Hx and Hx groups. CONCLUSION: Hepatocellular apoptosis plays vital roles in jaundice rats, and biliary decompression is more effective in treatment of patients with severe jaundice before operation.
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Apoptose/fisiologia , Icterícia Obstrutiva/patologia , Icterícia Obstrutiva/cirurgia , Animais , Bilirrubina/sangue , Caspase 3 , Caspases/metabolismo , Coledocostomia , Modelos Animais de Doenças , Hepatectomia , Icterícia Obstrutiva/sangue , Icterícia Obstrutiva/fisiopatologia , Testes de Função Hepática , Masculino , Ratos , Ratos Wistar , Fatores de TempoRESUMO
OBJECTIVE: To evaluate the results of palliative surgical treatment of hilar cholangiocarcinoma in terms of quality of life, survival period and cholangitis rate. METHODS: The clinical data on 232 patients with hilar cholangiocarcinoma in the last 22 years were analyzed retrospectively. Palliative operations included extrahepatic or intrahepatic choledochojejunostomy (123 patients), bridge internal drainage (15), endoscopic biliary drainage (49), percutaneous transhepatic biliary drainage or celiotomy biliary drainage (29), and exploratory celiotomy external drainage (16). RESULTS: In this series, the operative mortality rate was 9.1%, and no significant difference was observed between groups. The rate of cholangitis after operation was significantly lower in Roux-en-Y choledochojejunostomy group (16.2%) and bridge internal drainage group (15.4%) than in internal drainage group (35.5%, P<0.01), including percutaneous transhepatic biliary drainage (PTBD), endoscopic retrograde biliary drainage (ERBD), and celiotomy (or PTBD) external biliary drainage group (39.1%, P<0.01). No significant difference in survival was observed between the Roux-en-Y choledochojejunostomy group (9.3+/-1.8 months) and PTBD (or ERBD) internal drainage group (8.7+/- 2.2 months), but the survivals of the above groups were significantly longer than those of the bridge internal drainage group (6.5+/-1.7 months, P<0.05) and celiotomy (or PTBD) external biliary drainage group (4.4+/-2.1 months, P<0.01). CONCLUSIONS: In unresectable cholangiocarcinomas, either operative bilioenteric bypass or percutaneous transhepatic biliary drainage can achieve significant palliation. Roux-en-Y choledochojejunostomy is the best choice for palliative operation. The use of U-tube is recommended for internal radiation therapy.
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Neoplasias dos Ductos Biliares/cirurgia , Ductos Biliares Intra-Hepáticos/cirurgia , Colangiocarcinoma/cirurgia , Procedimentos Cirúrgicos do Sistema Digestório/métodos , Cuidados Paliativos/métodos , Adulto , Idoso , Idoso de 80 Anos ou mais , Neoplasias dos Ductos Biliares/complicações , Colangiocarcinoma/complicações , Colangite/epidemiologia , Colangite/etiologia , Procedimentos Cirúrgicos do Sistema Digestório/efeitos adversos , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Qualidade de Vida , Estudos Retrospectivos , Análise de SobrevidaRESUMO
OBJECTIVE: To clone gene fragments from suppression substructive library established for screening down-regulated genes in gastric carcinoma, and obtain the full-length novel gene. METHODS: Gene fragments were identified by sequencing plasmids of positive colonies chosen randomly. One gene fragment was amplified by RACE, and the full-length novel gene was obtained. Expression of novel gene mRNA was respectively detected by semi-quantitative PCR in the gastric carcinoma tissues and counterpart normal gastric mucous membrane of 25 patients with gastric cancer. The structure of the full-length novel gene, location on chromosome, property of protein encoded by full-length novel gene and its function were investigated by Bio-message technique. RESULTS: One 331 bp gene fragment was cloned, and its full-length novel gene obtained by RACE. The novel full-length gene was named GDDR, registered in the number of AF494509 by GenBank. Significant down-regulated expression of GDDR gene mRNA in gastric carcinoma tissues was confirmed (GDDR/beta-actin 13.474 +/- 5.059 vs 1.041 +/- 0.202, P < 0.01). GDDR was located in chromosome 2p13 with 5 exons. As one member of new BRICHO family as CA11, GDDR encoding protein with transmembrane peptide revealed homology to protein encoded by CA11. CONCLUSION: A novel full-length gene GDDR is obtained. GDDR likely is another gene of BRICHO family related to gastric cancer.
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Proteínas de Membrana/genética , Proteínas de Neoplasias/genética , Neoplasias Gástricas/genética , Sequência de Aminoácidos , Proteínas de Transporte , Mapeamento Cromossômico , Cromossomos Humanos Par 2/genética , Clonagem Molecular , DNA Complementar/química , DNA Complementar/genética , Regulação para Baixo , Regulação Neoplásica da Expressão Gênica , Humanos , Dados de Sequência Molecular , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Alinhamento de Sequência , Análise de Sequência de DNA , Homologia de Sequência de AminoácidosRESUMO
AIM: To investigate the expression of NGF family and their receptors in gastric carcinoma and normal gastric mucosa, and to elucidate their effects on gastric carcinoma. METHODS: RNA of gastric cancer tissues and normal gastric tissues was respectively isolated and mRNA was purified. Probes of both mRNA reverse transcription product cDNAs labeled with alpha-(33)P dATP were respectively hybridized with Atlas Array membrane where NGF and their family genes were spotted on. Hybridized signal images were scanned on phosphor screen with ImageQuant 5.1 software after hybridization. Normalized values on spots were analyzed with ArrayVersion 5.0 software. Differential expression of NGF family and their receptors mRNA was confirmed between hybridized Atlas Array membranes of gastric cancer tissues and normal gastric mucosa, then their effects on gastric carcinoma were investigated. RESULTS: Hybridization signal images on Atlas Array membrane appeared in a lower level of nonspecific hybridization. Both of NGF family and their receptors Trk family mRNA were expressed in gastric cancer and normal gastric mucosa. But adversely up-regulated expression in other tissues and organs. NGF, BDGF, NT-3, NT-4/5, NT-6 and TrkA, B and C were down-regulated simultaneously in gastric carcinoma in comparison with normal gastric mucosa. Degrees of down-regulation in NGF family were greater than those in their receptors Trk family. Down-regulation of NT-3 and BDGF was the most significant, and TrkC down-regulation level was the lowest in receptors Trk family. CONCLUSION: Down-regulated expression of NGF family and their receptors Trk family mRNA in gastric cancer is confirmed. NGF family and their receptors Trk family probably play a unique role in gastric cancer cell apoptosis by a novel Ras or Raf signal transduction pathway. Their synchronous effects are closely associated with occurrence and development of gastric carcinoma induced by reduction of signal transduction of programmed cell death.
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Carcinoma/genética , Fatores de Crescimento Neural/genética , Análise de Sequência com Séries de Oligonucleotídeos , Receptores de Fator de Crescimento Neural/genética , Neoplasias Gástricas/genética , Regulação Neoplásica da Expressão Gênica , Humanos , Hibridização de Ácido Nucleico , RNA Mensageiro/análiseRESUMO
AIM: To establish cDNA suppression subtraction library with a high subtraction efficiency by counterpart normal gastric mucosa mixture mRNA subtracting gastric cancer cells mixture mRNA for screening down-regulated genes in gastric carcinoma. METHODS: RNA of gastric cancer tissues and counterpart normal gastric mucosa were respectively isolated in five patients with gastric cancer, and their mRNA was purified. cDNA suppression subtraction library was established by counterpart normal gastric mucosa mixture mRNA (tester) subtracting gastric cancer tissues mixture mRNA (driver) of five patients with gastric carcinoma. The library plasmids were transformed into competent bacteria DH5a after ligation of the library cDNA fragments with T vectors. Library plasmids were extracted after picking colonies and shaking bacteria overnight. Its subtraction efficiency was confirmed by PCR and reverse hybridization of a nylon filter onto which the colonies of bacteria were transferred with probes of reverse transcription products cDNA of gastric cancer tissues mRNA and counterpart normal gastric mucosa mRNA labeled with alpha- (32)P dCTP. RESULTS: mRNA purified from total RNA of gastric cancer tissues and counterpart normal gastric mucosa in five patients with gastric carcinoma revealed a good quality. cDNA suppression subtraction library constructed for screening down-regulated genes in gastric carcinoma represented a high subtraction efficiency. 86 % of differential expression in down-regulated genes between counterpart normal gastric mucosa and gastric carcinoma was confirmed. CONCLUSION: cDNA suppression subtraction library with a high subtraction efficiency for screening down-regulated genes in gastric carcinoma is successfully established.