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1.
Chinese Journal of Biochemistry and Molecular Biology ; (12): 363-371, 2021.
Artigo em Chinês | WPRIM | ID: wpr-1015974

RESUMO

Krüppel-like factor 7 (KLF7) is a negative regulator of adipogenesis, whereas hypoxia-inducible factor 1 (HIF1) promotes anoxic-induced adipose tissue development in mammals. Our previous ChIP-seq analysis showed that one of the KLF7 binding peaks was present upstream of hypoxia-inducible factor 1 alpha (HIF1α), indicating that KLF7 may regulate HIF1α transcription. For this purpose, ChIP-PCR was used to verify ChIP-seq results, which showed that KLF7 directly bound to the HIF1α upstream region. Dual luciferase reporter and qRT-PCR results showed that KLF7 overexpression significantly decreased the luciferase reporter activity of HIF1α (- 4 432/- 4 182) (P < 0. 01) and inhibited HIF1α expression. After the deletion of KLF7 binding motif “TGCGCAGCAA” (- 4 300/-4 290) predicted by bioinformatics, the luciferase reporter activity of HIF1α (-4 432/-4 182) was significantly enhanced compared with wild-type plasmid (P<0. 01). Furthermore, Northeast Agricultural University broiler lines divergently selected for abdominal fat content (NEAUHLF) at the age of 1-7 weeks from the 19

2.
Chinese Journal of Applied Physiology ; (6): 5-8, 2019.
Artigo em Chinês | WPRIM | ID: wpr-776569

RESUMO

OBJECTIVE@#To investigate the expression of EGR1 gene and the localization of EGR1 protein in bovine skeletal muscle-derived satellite cells (MDSCs), as well as to investigate the mechanism that EGR1 protein enters the nucleus.@*METHODS@#Bovine MDSCs were cultured in differentiation medium for 1 day, 3 days and 5 days, respectively, and each group was triplicate. The expression of EGR1 gene and the localization of EGR1 protein were studied at different differentiation period in MDSCs by qRT-PC and Western blot. Moreover, the changes on the expression of endogenous EGR1 gene and EGR1 proteins were explored by CRISPRi, site-directed mutagenesis and laser confocal method.@*RESULTS@#The results from the qRT-PCR and Western blot showed that the expressions of EGR1 gene on transcription level and translation level were significantly higher in differentiated cells than those in undifferentiated cells. The highest expression was found on the third day after the differentiation, and then began to decline. Immunofluorescence assays showed that EGR1 proteins were preferentially expressed in differentiated MDSCs, and increased along with the increase of number of myotubes. Confocal observation revealed that some EGR1 proteins were transferred into the nucleus in the differentiation of cells, however, the EGR1 proteins would not be detected in the differentiated MDSCs nuclei if a site directed mutagenesis (serine) on EGR1 protein occurred.@*CONCLUSION@#During the differentiation of bovine skeletal muscle satellite cells, the transcriptional level of EGR1 gene is increased, and some EGR1 proteins are transferred into the nucleus. The serine phosphorylation at position 533 of the C terminal of EGR1 protein is necessary for the nucleus transfer.


Assuntos
Animais , Bovinos , Diferenciação Celular , Núcleo Celular , Células Cultivadas , Proteína 1 de Resposta de Crescimento Precoce , Genética , Metabolismo , Fibras Musculares Esqueléticas , Células Satélites de Músculo Esquelético , Metabolismo
3.
Acta Physiologica Sinica ; (6): 379-386, 2012.
Artigo em Chinês | WPRIM | ID: wpr-333188

RESUMO

This paper was aimed to study conserved motifs of voltage sensing proteins (VSPs) and establish a voltage sensing model. All VSPs were collected from the Uniprot database using a comprehensive keyword search followed by manual curation, and the results indicated that there are only two types of known VSPs, voltage gated ion channels and voltage dependent phosphatases. All the VSPs have a common domain of four helical transmembrane segments (TMS, S1-S4), which constitute the voltage sensing module of the VSPs. The S1 segment was shown to be responsible for membrane targeting and insertion of these proteins, while S2-S4 segments, which can sense membrane potential, for protein properties. Conserved motifs/residues and their functional significance of each TMS were identified using profile-to-profile sequence alignments. Conserved motifs in these four segments are strikingly similar for all VSPs, especially, the conserved motif [RK]-X(2)-R-X(2)-R-X(2)-[RK] was presented in all the S4 segments, with positively charged arginine (R) alternating with two hydrophobic or uncharged residues. Movement of these arginines across the membrane electric field is the core mechanism by which the VSPs detect changes in membrane potential. The negatively charged aspartate (D) in the S3 segment is universally conserved in all the VSPs, suggesting that the aspartate residue may be involved in voltage sensing properties of VSPs as well as the electrostatic interactions with the positively charged residues in the S4 segment, which may enhance the thermodynamic stability of the S4 segments in plasma membrane.


Assuntos
Arginina , Química , Ácido Aspártico , Química , Membrana Celular , Fisiologia , Sequência Conservada , Ativação do Canal Iônico , Canais Iônicos , Química , Potenciais da Membrana , Estrutura Terciária de Proteína
4.
Chinese Journal of Applied Physiology ; (6): 105-108, 2010.
Artigo em Chinês | WPRIM | ID: wpr-340222

RESUMO

<p><b>OBJECTIVE</b>To study the effect of chronic rapid eye movement sleep deprivation on energy metabolism, FT3, FT4 in serum.</p><p><b>METHODS</b>Rapid eye movement sleep deprivation of rats were deprived by flower pot, and then the energy metabolism were detected. The FT3, FT4 level in serum was determined by radioimmunoassay kit.</p><p><b>RESULTS</b>Rats after sleep deprivation displayed food intake increased from (75.06 +/- 25.37)g/(d x kg) to (122.30 +/- 20.43)g/(d x kg), body weight substantially decreased from (360.89 +/- 43.01) g to (295.97 +/- 37.95) g, body temperature from (37.62 +/- 1.12) degrees C up to the first (39.00 +/- 0.87) degrees C and then reduced to (37.72 +/- 0.84) degrees C, the basal metabolism rate increased significantly from (1.69 +/- 0.36) mlO2/(g x h) to (2.40 +/- 0.09) mlO2/(g x h), compared with the control group( P < 0.05). Sleep deprivation also resulted significantly lower serum thyroxine levels in comparison with the control, serum free triiodothyronine (FT3) level reduced from (3.38 +/- 0.88) pmol/L to (2.38 +/- 0.83) pmol/L, then free thyroxine(FT4) decreased from (14.62 +/- 3.62) pmol/L to (8.26 +/- 2.80) pmol/L (P < 0.05).</p><p><b>CONCLUSION</b>Rapid eye movement sleep deprivation can change energy metabolism remarkable, as well as the alteration of FT3, FT4 levels in serum.</p>


Assuntos
Animais , Masculino , Ratos , Metabolismo Energético , Fisiologia , Ratos Wistar , Privação do Sono , Sangue , Metabolismo , Sono REM , Fisiologia , Tiroxina , Sangue , Fatores de Tempo , Tri-Iodotironina , Sangue
5.
Chinese Journal of Cancer ; (12): 25-29, 2010.
Artigo em Chinês | WPRIM | ID: wpr-292645

RESUMO

<p><b>BACKGROUND AND OBJECTIVE</b>Epithelial ovarian cancer involves a number of factors. Recent studies have shown that osteopontin (OPN) is related to the occurrence and development of a variety of tumors, but few studies are on ovarian cancer. B7-H4 is a newly identified tumor marker in ovarian cancer. This study explored the expression of OPN and B7-H4 and their clinical significance in epithelial ovarian tumors.</p><p><b>METHODS</b>The expression of OPN and B7-H4 in 15 cases of normal ovarian tissue, 20 of benign ovarian tumor tissue, 20 of borderline ovarian tumor tissue, and 40 of ovarian cancer tissue were detected by immunohistochemistry, and the relationship of OPN and B7-H4 expression to clinical and pathologic features of ovarian cancer was analyzed.</p><p><b>RESULTS</b>The expression of OPN and B7-H4 were significantly higher in ovarian cancer than in borderline and benign tumors (P<0.05). The positive rates of OPN and B7-H4 were significantly higher in poorly differentiated ovarian cancer than in medium and highly differentiated ovarian cancer (P<0.05), and the levels of expression were significantly lower in tissue at stages I and III of ovarian cancer than in stages III and IV (P<0.05). The positive rate of OPN associated with a higher rate of lymph node metastasis (P<0.05), but did not relate to age and histologic type. The positive rate of B7-H4 were significantly higher in ovarian serous carcinoma than in the mucinous carcinoma (P<0.05), but did not relate to age and lymph node metastasis.</p><p><b>CONCLUSION</b>The expression of OPN and B7-H4 increased in epithelial ovarian cancer, which could be referenced in the diagnosis of ovarian malignant tumors.</p>


Assuntos
Adolescente , Adulto , Idoso , Feminino , Humanos , Pessoa de Meia-Idade , Adulto Jovem , Adenocarcinoma Mucinoso , Diagnóstico , Metabolismo , Patologia , Cistadenocarcinoma Seroso , Diagnóstico , Metabolismo , Patologia , Regulação Neoplásica da Expressão Gênica , Metástase Linfática , Estadiamento de Neoplasias , Neoplasias Epiteliais e Glandulares , Diagnóstico , Metabolismo , Patologia , Osteopontina , Metabolismo , Neoplasias Ovarianas , Diagnóstico , Metabolismo , Patologia , Ovário , Metabolismo , Inibidor 1 da Ativação de Células T com Domínio V-Set , Metabolismo
6.
Chinese Journal of Oncology ; (12): 697-700, 2007.
Artigo em Chinês | WPRIM | ID: wpr-298515

RESUMO

<p><b>OBJECTIVE</b>To investigate the expression of GLUT1, p63 and DNA-Pkcs in serous ovarian tumors and their significance.</p><p><b>METHODS</b>GTUL1, p63 and DNA-Pkcs expression at protein level was detected by immunohistochemistry in patients with serous ovarian tumors. Chi-square analysis was used to assess if their expression is associated with clinicopathologic characteristics of the tumors.</p><p><b>RESULTS</b>Cells in the normal ovarian tissues were not stained with GTUL1 and p63 antiserum, but DNA-Pkcs was positively stained. The intensity of GTUT1 and p63 expression was stronger in malignant ovarian serous tumors compared with other subtypes (P < 0.01). There were significant differences of DNA-PKcs among normal ovaries (100.0%), benign (95.0%), borderline (90.0%) and malignant (60.0%) serious ovarian neoplasms (P < 0.01). The level of GLUT-1 expression was correlated with FIGO staging, intraperitoneal implantation, ascites and lymph node metastasis (P < 0.05). p63 expression was associated with clinicopathologic characteristics except ascites (P < 0.05). DNA-PKcs was only correlated with FIGO staging and lymph node metastasis (P < 0.05).</p><p><b>CONCLUSION</b>The results suggest that the abnormal expression of GTUT1, p63 and DNA-Pkcs may perhaps participate in serous ovarian tumor occurrence and development and may be considered as a marker reflecting tumor malignant behavior.</p>


Assuntos
Adolescente , Adulto , Idoso , Feminino , Humanos , Pessoa de Meia-Idade , Adulto Jovem , Cistadenocarcinoma Seroso , Metabolismo , Patologia , Cistadenoma Seroso , Metabolismo , Patologia , Proteína Quinase Ativada por DNA , Metabolismo , Epitélio , Metabolismo , Regulação Neoplásica da Expressão Gênica , Transportador de Glucose Tipo 1 , Metabolismo , Imuno-Histoquímica , Metástase Linfática , Estadiamento de Neoplasias , Proteínas Nucleares , Metabolismo , Neoplasias Ovarianas , Metabolismo , Patologia , Ovário , Biologia Celular , Transativadores , Metabolismo , Fatores de Transcrição , Proteínas Supressoras de Tumor , Metabolismo
7.
Cancer Research and Clinic ; (6)2006.
Artigo em Chinês | WPRIM | ID: wpr-676536

RESUMO

Objective To study the expression and function relations of the p63 in ovarian serous carcinoma,and compare them with normal ovary tissue.Methods RT-PCR,in situ hybridization and im- munohistochemistry were used to detect the expression of the p63 in ovarian serous carcinoma,ovarian bor- derline serous tumors,ovarian benign serous tumors and normal ovary tissues.Results All samples ex- pressed p63 mRNA through RT-PCR and immunohistochemistry,while the levels of expression in ovarian tu- mors was higher than that of normal ovary.p63 in human borderline and malignant ovarian tumors was higher than that in human benign ovarian tumors with significant difference,which suggested p63 may play great roles in the origin and progression of ovarian tumors.The result of in situ hybridization showed that positive expression rate of p63 mRNA in ovarian cancer (92.1%) was significantly higher than that in normal ovary(0) or LWP ovarian cancer(50.0%).Conclusion Expression of p63 plays an important role in the development of varian carcinoma,and along with the difference of histological stage and clinico-pathological stage,the ex- pression of p63 gene and protein level were different.

8.
Chinese Journal of Epidemiology ; (12): 15-18, 2006.
Artigo em Chinês | WPRIM | ID: wpr-295614

RESUMO

<p><b>OBJECTIVE</b>To evaluate visual inspection with Lugol's iodine (VILI) in cervical cancer screening program and to provide evidence for designing a cervical cancer screening algorithm in high risk areas of existing low-resource settings to reduce the incidence and mortality of cervical cancer.</p><p><b>METHODS</b>Women in Yangcheng county, Shanxi province were screened with VILI, colposcopy, liquid-based cytology test and human papilloma virus (HPV) DNA test. The efficacy of different screening tests was compared by Youden's index based on the pathology as the gold standard.</p><p><b>RESULTS</b>In the population being screened, the mean age was 40.80 +/- 10.75 years old. Based on pathological findings, 4.35% (32/735) of the subjects had >or= CIN (cervical intraepithelial neoplasia) II. The sensitivity and specificity for the VILI test (>or= positive) were 53.13 and 82.19, while 56.25 and 79.09 were for colposcopy (>or= low grade dysplasia) respectively. Comparing by the Youden's indexs, there was no statistically significant difference (P > 0.05) between VILI and colposcopy. However, statistical significant difference (P < 0.05) was found between VILI and liquid-based cytology test and HPV DNA test. In addition, there was no statistically significant difference (P > 0.05) found between the experienced doctors and the newly-trained doctors working in the field station.</p><p><b>CONCLUSION</b>With low sensitivity when using microscope but low cost of equipments, VILI can be one of the primary screening tests in China's rural area with low-resource settings if the screening frequency is to be increased.</p>


Assuntos
Adulto , Feminino , Humanos , Displasia do Colo do Útero , Diagnóstico , Epidemiologia , China , Epidemiologia , Detecção Precoce de Câncer , Economia , Métodos , Recursos em Saúde , Incidência , Iodetos , Avaliação de Programas e Projetos de Saúde , Saúde da População Rural , Sensibilidade e Especificidade , Fatores Socioeconômicos , Neoplasias do Colo do Útero , Diagnóstico , Epidemiologia
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