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1.
Chongqing Medicine ; (36): 2261-2265, 2018.
Artigo em Chinês | WPRIM | ID: wpr-692087

RESUMO

Objective To explore the effect of oridonin on the endoplasmic reticulum stress (ERS) in colonic epithelium of ulcerative colitis (UC) mice.Methods The UC mice model was established by sodium dextran sulfate (DSS),and the intervention group of oridonin and sulfasalazine was set up,the disease activity index (DAD was measured,the colonic tissue was evaluated by histopathologidscore (HPS),and RT-qPCR was used to detect the expression of inflammatory cytokines tumor factor-α (TNF-α),interleukin 6 (IL-6),cyclooxygenase 2 (COX-2),glucose-regulated protein 78 (GRP78),transcription factor EBP homologous protein (CHOP),activator transcription factor 6 (ATF6),protein kinase R like endoplasmic reticulum kinase (PERK) and inositol requiring enzyme 1α (IRE1α).Results The expression of TNF-α,IL-6,COX-2,GRP78,ATF6,CHOP,PERK and IRE1α mRNA in the colonic epithelium of the model group were all up-regulated obviously as compared with the healthy control group(P<0.05,P<0.01).When compared with the model group,DAI and HPS in oridonin-treated group were significantly decreased (P<0.05,P<0.01),which the curative effect was similar to that of the sulfasalazine group(P>0.05,P<0.01).The expression of TNF-α,IL-6,COX-2,GRP78,CHOP,ATF6 and PERK mRNA levels were significantly reduced in oridonin-treated group(P< 0.05,P<0.01).Conclusion Oridonin can alleviate colonic inflammation induced by DSS and its mechanism may be related to ERS of colonic epithelial tissue.

2.
Artigo em Chinês | WPRIM | ID: wpr-239192

RESUMO

<p><b>OBJECTIVE</b>To investigate the effect of mTOR signal transduction pathway and down-regulating anti-oncogene PTEN on the growth of breast cancer MCF-7 cells.</p><p><b>METHODS</b>MCF-7 cells were transfected with the eukaryotic expression plasmid pcDNA3.1-mTOR and non-loaded plasmid, and the expression of mTOR in the cells was detected using Western blotting. Flow cytometry was used to analyze apoptosis and cell cycle of the transfected cells, and the expression of PTEN was detected after transfection.</p><p><b>RESULTS</b>The cells transfected with pcDNA3.1-mTOR showed a increased growth rate than those transfected with the non-loaded plasmid and those without transfection. The expression of the protein PTEN decreased obviously in the cells after mTOR trasnfection.</p><p><b>CONCLUSION</b>mTOR can regulate the expression of PTEN via PI3K/AKT/PTEN pathways through a negative feedback mechanism. Increased mTOR expression promotes MCF-7 cell growth, suggesting the potential value of mTOR specific inhibitor in the treatment of breast cancer.</p>


Assuntos
Humanos , Apoptose , Neoplasias da Mama , Patologia , Ciclo Celular , Regulação para Baixo , Regulação Neoplásica da Expressão Gênica , Células MCF-7 , PTEN Fosfo-Hidrolase , Metabolismo , Plasmídeos , Transdução de Sinais , Serina-Treonina Quinases TOR , Genética , Transfecção
3.
Artigo em Chinês | WPRIM | ID: wpr-355245

RESUMO

<p><b>OBJECTIVE</b>To modify the level set method for precise and fast segmentation of B-type ultrasound image lesions.</p><p><b>METHODS</b>Based on the best of region level set method, entropy in the information theory was introduced into image processing to define a dynamic weighting factor that responded to the gradient change of the local gray levels to evaluate the dynamic degree of driven force on each pixel on the contour to the target and background areas. The dynamic weighting factors were reconciled into the regional level set model and led the contour to deform and move during the iterations. As lesion segmentation was classified as local segmentation of a specific area, the calculation was restrained to the local sphere abide by the contour, which greatly reduced the calculation complex.</p><p><b>RESULTS</b>Experiments on B-type ultrasound images showed improved results of the proposed method with a better accuracy and less time consumption compared with several current level set methods.</p><p><b>CONCLUSION</b>Level set method reconciled with dynamic weighting factor allows a better evaluation of the lesion contour pixels, and the local calculation strategy results in an enhanced segmentation efficiency.</p>


Assuntos
Algoritmos , Processamento de Imagem Assistida por Computador , Modelos Teóricos , Ultrassonografia
4.
Artigo em Chinês | WPRIM | ID: wpr-249354

RESUMO

<p><b>OBJECTIVE</b>To screen differentially expressed genes and identify potential signaling pathway in Asian people with breast cancer.</p><p><b>METHODS</b>Five gene microarray datasets of Asian people with breast cancer, GSE6367, GSE9309, GSE15852, GSE33447 and GSE45255, were downloaded from GEO. Microarrays with 318 breast cancer and 60 normal breast tissues were used for analysis of differentially expressed genes and pathway. 32 pairs of breast cancer patients' specimens were used to validate the differentially expressed genes by real-time PCR.</p><p><b>RESULTS</b>Analysis of the large sample of microarray data identified 436 differentially expressed genes in breast cancer tissues, while 259 of these genes were up-regulated and the other 177 down-regulated. Pathway analysis showed that metabolism-related signaling pathway may be involved in the development of breast cancer in Asian people. The expressions of KRT19, ADIPOQ, CFD, RBP4, LPL, ABCA8 and CD36 genes were confirmed by real-time PCR.</p><p><b>CONCLUSION</b>This study shows differential gene expression profile and potential signaling pathway in Asian people with breast cancer. CD36 gene may be closely related to the Asian breast cancer. ABCA8 gene may be a new disease gene in Asian breast cancer.</p>


Assuntos
Feminino , Humanos , Povo Asiático , Neoplasias da Mama , Genética , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Análise de Sequência com Séries de Oligonucleotídeos , Reação em Cadeia da Polimerase em Tempo Real , Transdução de Sinais , Transcriptoma
5.
Artigo em Chinês | WPRIM | ID: wpr-312636

RESUMO

Axillary interventions, represented by axillary lymph node dissection (ALND), has been a key component in radical surgeries for breast cancer since the proposal of radical mastectomy. ALND substantially affects the quality of life of the patients. In recent years, many studies suggested that axillary interventions may not be necessary for all breast cancer patients, and for early early-stage patients, it brings more harm than benefit. Sentinel lymph node biopsy (SLNB) has provided good guidance to indicate the necessity of ALND, but several studies revealed that not all patients with positive SLNB results benefited from ALND. In this review, the authors summarize the recent progress of researches of these two hot issues.


Assuntos
Feminino , Humanos , Axila , Neoplasias da Mama , Cirurgia Geral , Excisão de Linfonodo , Linfonodos , Qualidade de Vida , Biópsia de Linfonodo Sentinela
6.
Artigo em Chinês | WPRIM | ID: wpr-447489

RESUMO

Objective:This study aimed to establish a mouse model of breast cancer by inoculating human breast cancer cells into mice with normal immune function. Methods:Forty female BALB/C mice were randomized into four groups, with 10 mice in each group. The four groups were established according to the dosage of cyclophosphamide and prednisone, namely, the control group, low dose group, medium dose group, and high dose group. The mouse models of breast cancer were established by injecting human breast cancer cells into the fat pad of the right second breast of mice in the groups. Mice in the four groups were observed based on the time of tumorigenesis, rate of tumor formation, tumor imaging and pathological features, and metastasis of vital internal organs. Results:In the high dose group, the time of tumor formation was lower than that of the other groups, but the rate of tumor formation was high. Some visceral metastases occurred in the mice. By contrast, the medium dose group revealed completely opposite results. No death and tumor formation in both the control and low dose groups were reported. Conclusion:A human breast carcinoma model in mice was successfully established. Using this model, the onset and development of breast cancer could be much better imitated in the normal immune system of mice.

7.
Artigo em Inglês | WPRIM | ID: wpr-634782

RESUMO

Hypermethylation in the promoter region is an important epigenetic mechanism for the transcriptional repression of a number of cancer-associated genes, and over-expression and/or increased activity of DNA methyltransferases are considered to be the main cause of promoter hypermethylation. In order to explore the roles of two methyltransferase members (DNMT1 and DNMT3b) in the cholangiocarcinoma tumorigenesis, antisense eukaryotic expression plasmid of DNMT1 and DNMT3b gene was constructed respectively, and were co-transfected into the human cholangiocarcinoma cell line QBC-939 to observe their biological effects on the cell growth and proliferation ability, apoptosis, cell cycle alteration, and the tumorigenesis ability in the subcutaneous tissue of nude mouse. The results demonstrated that co-transfection with antisense eukaryotic expression plasmid of DNMT1 and DNMT3b gene and single transfection with antisense eukaryotic expression plasmid of DNMT1 gene can suppress the growth and proliferation of QBC-939, block the cell cycle at G1 phase, increase the apoptosis rate, minimize the tumor size in the subcutaneous tissue of nude mouse. The suppressing biological effect of co-transfection is stronger than single transfection with antisense DNMT1. Meanwhile, single transfection with antisense eukaryotic expression plasmid of DNMT3b gene has no effects on the biological characteristics of QBC-939. This study suggests that DNMT1 gene plays a key role in DNA methylation and DNMT3b gene may act as an accessory to support its function in inactivation of tumor suppressor genes. Combination DNMT1 and DNMT3b will increase their biological effects and have the synergistic effect on suppressing the growth of human cholangiocarcinoma cell line QBC-939.


Assuntos
Apoptose , Neoplasias do Sistema Biliar/metabolismo , Linhagem Celular Tumoral , Proliferação de Células , Colangiocarcinoma/metabolismo , DNA (Citosina-5-)-Metiltransferases/biossíntese , DNA (Citosina-5-)-Metiltransferases/genética , Regulação para Baixo , Regulação Neoplásica da Expressão Gênica , Vetores Genéticos , Camundongos Nus , Transplante de Neoplasias
8.
Artigo em Chinês | WPRIM | ID: wpr-234549

RESUMO

Hypermethylation of the promoter region is one of the major mechanism of tumor suppressor gene inactivation. In order to provide a research tool for the study on the function of MBD1gene in DNA methylation and tumorigenesis, antisense MBD1 gene eukaryotic expression plasmid was constructed and transfected into human biliary tract carcinoma cell line QBC-939 to observe its effect on the expression of MBD1 mRNA and protein by using RT-PCR and FCM respectively. Following the transfection, the mRNA level of MBD1 gene decreased from 0. 912±0. 022 to 0. 215±0.017, and the protein level of MBD1 gene also decreased from (80.19±5.05) % to (35.11±4.05)%. There were very significant differences in the expression both at the transcription and post-transcription levels of MBD1 gene between non-tranfection group and the antisense MBD1 gene eukary otic expression plasmid transfection group (P<0.01). It was suggested that transfection with the antisense MBD1 gene eukaryotic expression plasmid can significantly reduce the expression level of MBD1 gene in QBC-939, and this study may provide a valid tool for the investigation of the function of MBD1 gene and its role in biliary tract carcinoma.

9.
Artigo em Inglês | WPRIM | ID: wpr-634286

RESUMO

Hypermethylation of the promoter region is one of the major mechanism of tumor suppressor gene inactivation. In order to provide a research tool for the study on the function of MBD1 gene in DNA methylation and tumorigenesis, antisense MBD1 gene eukaryotic expression plasmid was constructed and transfected into human biliary tract carcinoma cell line QBC-939 to observe its effect on the expression of MBD1 mRNA and protein by using RT-PCR and FCM respectively. Following the transfection, the mRNA level of MBD1 gene decreased from 0. 912 +/- 0.022 to 0.215 +/- 0. 017, and the protein level of MBD1 gene also decreased from (80.19 +/- 5.05) % to (35.11 +/- 4.05) %. There were very significant differences in the expression both at the transcription and post-transcription levels of MBD1 gene between non-tranfection group and the antisense MBD1 gene eukaryotic expression plasmid transfection group (P < 0.01). It was suggested that transfection with the antisense MBD1 gene eukaryotic expression plasmid can significantly reduce the expression level of MBD1 gene in QBC-939, and this study may provide a valid tool for the investigation of the function of MBD1 gene and its role in biliary tract carcinoma.


Assuntos
Neoplasias do Sistema Biliar/metabolismo , Neoplasias do Sistema Biliar/patologia , Linhagem Celular Tumoral , Metilação de DNA , Proteínas de Ligação a DNA/biossíntese , Proteínas de Ligação a DNA/genética , Células Eucarióticas/metabolismo , Regulação Neoplásica da Expressão Gênica , Vetores Genéticos , Oligonucleotídeos Antissenso/genética , Plasmídeos/genética , Fatores de Transcrição/biossíntese , Fatores de Transcrição/genética , Transfecção
10.
Artigo em Chinês | WPRIM | ID: wpr-559844

RESUMO

Objective:To investigate the influences of IGF-1 on the stress homone,InsR?、? gene expression and protein content in pyaemia rats.Methods:SD rats with abdominal infection models established by cecal ligation and perforation method were randomly divided into three groups: pyaemic group(n=10),parenteral nutrition group(PN group,n=10) and IGF-1 Group (n=10),and 10 SD rats were used as normal group(n=10).On the sixth day,Vena Cava blood was sampled to determine the level of glucose,insulin,glucagon,and IGF-1.In addition,the expression of the InsR?、? in liver and muscle were detected by immunohistochemistry and Real-Time PCR methods.Results:The plasma glucose,insulin,glucagon in the IGF-1 group were significantly lower than that of the pyaemic group and PN group(P

11.
Artigo em Chinês | WPRIM | ID: wpr-543901

RESUMO

Objective To study the effects of glutamine (Gln) combined with growth hormone (GH) on the levels of cytokine (TNF-?, IL-1, IL-6), coritsol and amino acid metabolism in septic rats. Methods Ten out of 79 SD rats were randomly collected as the control group. Thirty of 69 septic SD rats, which were made by cecal ligation and perforation (CLP) method and were given parenteral nutrition (PN) lived to day 6. They were also randomly divided into three groups as follows: septic group (n=10), parenteral supplemented glutamine group (Gln group, n=10), and Gln combined with GH (Gln+GH group, n=10). On the 6th day, blood drew from portal veins of the dead rats was used to detect the levels of TNF-?, IL-1, IL-6 and cortisol by ELISA. The plasma concentrations of free amino acids were determined by amino acid auto-analyzer. The muscle tissue of extensor digitorum longus was used to determine 3-methyl-histidine (3-MH) by high performance liquid chromatographic (HPLC). Results Except for the control group, most rats developed celiac abscess, hepatic abscess and pulmonary infection. The serum levels of TNF-?, IL-1, IL-6 and cortisol were significantly higher in the septic group than those of the other three groups, and they were significantly lower in the Gln+GH group than those of the Gln group, P

12.
Artigo em Chinês | WPRIM | ID: wpr-526392

RESUMO

Objective To study the effect of transfection of antisense MBD1 gene eukaryotic expression vector on the expression of MBD1 gene in human cholangiocarcinoma cell line QBC-939.Methods The(constructed) antisense MBD1 gene eukaryotic expression vector was transfected into the human(cholangiocarcinoma) cell line QBC-939 using lipofectamine transfection reagents,and positive cell clones were obtained using G418 selection after transfection.The constructed recombinant vector was transfected into(QBC-939) cells successfully and was confirmed by amplifying the exogenous neo~R gene with PCR method.The expression level of MBD1 gene mRNA and protein was detected by RT-PCR and FCM methods respectively.Results Following the transfection,the MBD1 gene mRNA level in human cholangiocarcinoma cell line QBC-939 decreased from 0.912?0.022 to 0.215?0.017,and the MBD1 gene protein level also(decreased) from(80.19?5.05)% to(35.11?4.05)%.There were very significant differences on the expression both at the transcription and post-transcription levels of MBD1 gene between non-tranfection group and the antisense MBD1 gene eukaryotic expression vector transfection group(P

13.
Artigo em Chinês | WPRIM | ID: wpr-527451

RESUMO

Objective To explore the expression and relationship of PTEN and mTOR in the development of cholangiocarcinoma,determine the effect of the PI3K/PTEN/AKT/mTOR signal transduction on the(development) of cholangiocarcinoma.Methods The expression of mTOR and PTEN in the cholangiocarcinoma was detected by the immunohistochemistry and RT-PCR method.Results Compared to normal tissues,the(expression) of mTOR gene in cholangiocarcinoma significantly increased,but the expression of PTEN gene(decreased).There was a negative correlation between mTOR gene and PTEN gene expression in(cholangiocarcinoma). Conclusions The expression of mTOR gene in cholangiocarcinoma was increased and the expression of PTEN was decreased.It suggested that the mTOR gene and PTEN gene could play an important role in the process of development of cholangiocarcinoma.

14.
Artigo em Chinês | WPRIM | ID: wpr-558303

RESUMO

Objective: To investigate the influences of IGF-1 on the stress hormone,GLUT-4 and its mRNA expression.Methods: The SD rats with abdominal infection established by cecal ligation and perforation method were randomly divided into three groups: pyaemic group(n=10),parenteral nutrition group(PN group,n=10) and IGF-1 Group(n=10).10 healthy rats was used as the normal group(n=10).On the sixth day,blood was sampled to determine the level of glucose,insulin,glucagon and IGF-1.The expressions of GLUT-4 and its mRNA in muscle were detected by immuno-histochemistry and Real-Time PCR methods.Results: The plasma levels of glucose,insulin and glucagon in the IGF-1 group were significantly lower than those in the pyaemic group and PN group(P

15.
Artigo em Chinês | WPRIM | ID: wpr-524370

RESUMO

Objective To investigate the effect of enteral and parenteral nutrition on gut microecology in rats with abdominal infection. Methods Fourteen Sprague-Dawley (SD) model rats of abdominal infection, which had survived for more than 6 days were divided into two groups: PN group ( n =7) and PN+EN group ( n =7) via jejunostomy and jugular vein respectively for another 5 days. The nutrition support in the two groups was isonitrogen and isocaloric. At sacrifice on the sixth day, occludin and IgA level in plasma cells of intestine epithelium of the gut were measured by immunohistochemistry. Vena cava blood and homogenated tissue of liver, lung and mesenteric lymph nodes were cultured to determine bacterial translocations, and portal vein blood was tested for endotoxin. Results The expression of occludin and IgA in the small and large intestine in PN+EN group were stronger than PN group ( P

16.
Artigo em Chinês | WPRIM | ID: wpr-527148

RESUMO

Objective To investigate the effects of the tumor suppressor gene PTEN in growing inhibition and down-regulating mTOR in cholangiocarcinoma QBC939 cells in vitro.Methods QBC939 cells were transfected with plasmids wild-type PTEN and C124S-PTEN in vitro.After transfection,the expression of the PTEN and phosphorylation of AKT and mTOR was detected by Western blot.Flow cytometry was used to analyze apoptosis and cell cycle of the transfected cells.Results Compared with the control,the expression of phosphorylation AKT was decreased and mTOR were down-regulated respectively when transfected with the wild-type PTEN.However,after transfection with mutation-type PTEN,the level of PTEN in the cells by increased,but phosphorylation AKT level and mTOR expression had no significant change.Conclusions PTEN can be actived by phosphorylated AKT.Actived AKT decreased the mTOR which led to tumor cells apoptosis and regulation of the tumor cell cycle.In the pathway of signal transmission of PI3K/AKT/PTEN/mTOR,PTEN and mTOR are closely related through phosphorylation of AKT.

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