Silence Research Articles

An Agent-based Model for an Act of Expression or Silence: Focused on College Students’ Behavior in Class

An Agent-based Model for an Act of Expression or Silence: Focused on College Students’ Behavior in Class

Philip Roth in His Silence

Philip Roth in His Silence

Breaking the Silence of Bathsheba: Imagining Younghye and Aphrodite Taking off the Patriarchy

Breaking the Silence of Bathsheba: Imagining Younghye and Aphrodite Taking off the Patriarchy

Poéticas experimentales en la resistencia cultural vasca: 'Isturitzetik Tolosan Barru' (1969)

Durante las décadas de los 60 y los 70, tras largos años de represión y silencio, se crearon movimientos populares que establecieron las bases de una red social y cultural, y contribuyeron a la estabilización y reconfiguración de la comunidad cultural vasca. Este artículo ofrece una aproximación a ese contexto sociocultural mediante el análisis detallado del primer libro de artista publicado en euskera: Isturitzetik Tolosan barru (Itxaropena, 1969) de JosAnton Artze. Para comprender tanto el contexto en el que emerge este poemario, como su aportación a la poética experimental, por una parte, se presentan las dinámicas colectivas y participativas que propiciaron la aparición de este primer libro artístico y colectivo; por otra parte, se analiza cómo se materializa lo experimental en este libro artístico. En ese contexto, se presenta un análisis textual y contextual de este poemario, incidiendo en su dimensión sonora, visual y arquitectónica. Finalmente, se reflexiona sobre cómo han tratado los discursos historiográficos de la literatura vasca esta aportación poética.

PFKFB3促进腺样囊性癌ACC-2细胞增殖、迁移、侵袭和转移

Objective To evaluate the expression of 6-phosphofructo-2-kinase/fructose-2, 6-bisphosphatase 3 (PFKFB3) in adenoid cystic carcinoma and the effects of PFKFB3 on the proliferation, migration, invasion and tumorigenesis in vivo of adenoid cystic carcinoma cells. Methods Immunohistochemistry and Western blotting were applied to detect the expression of PFKFB3 in 29 cases of adenoid cystic carcinoma tissues and 10 cases of para-carcinoma normal salivary gland tissues. PFKFB3 in adenoid cystic carcinoma ACC-2 cells was silenced by adenovirus vector. The effects of PFKFB3 silence on cell proliferation, migration, invasion and tumorigenesis in vivo and distant metastasis of adenoid cystic carcinoma cells were evaluated by cell proliferation assay, wound healing assay, Transwell assay, xenografted mice model and lung metastasis mice model. Results The results of immunohistochemical staining showed that the positive expression rate of PFKFB3 in adenoid cystic carcinoma tissues was 93.1% (27/29), and the positive expression rate of PFKFB3 in normal salivary gland tissues was 20.0% (2/10), with a significant difference (χ2=20.84, P<0.001). The results of methyl thiazolyl tetrazolium (MTT) assay showed that, compared with ACC-2 group, the proliferation activity of cancer cells with silence of PFKFB3 (PFKFB3--ACC-2) was significantly suppressed for 72 h (1.8±0.2 vs. 4.7±0.8, t=3.582, P=0.001). The results of wound healing assay showed that after scraping cells away for 24 h, the number of cells in the scratch area was 99.8±13.2 in the PFKFB3--ACC-2 group, which was significantly less than that in the ACC-2 group (263.0±97.4, t=2.868, P=0.029). Transwell results indicated that the number of cells passing through matrigel was 17.6±2.1 in the PFKFB3--ACC-2 group, which was less than that in the ACC-2 group (28.6±3.8, t=4.452, P=0.004). The tumor volume in the PFKFB3--ACC-2 [(623.5±134.1) mm3] was smaller than that in the ACC-2 group [(1 621.0±278.1) mm3,t=4.213, P=0.001]. More pulmonary metastases were found in the ACC-2 group than PFKFB3--ACC-2 group (18.1±3.2 vs. 4.1±2.2, t=6.322, P=0.001). Conclusion The expression of PFKFB3 is higher in adenoid cystic carcinoma tissue than normal salivary gland tissue, and the highly expressed PFKFB3 plays a driving role in the proliferation, migration, invasion and metastasis in adenoid cystic carcinoma. Key words: Carcinoma, adenoid cystic; Glycolysis; PFKFB3; Tumor development

MiR-205-3p、miR-205-5p对头颈鳞状细胞癌细胞增殖、迁移及侵袭的影响

Objective To investigate the effects of microRNA-205-3p (miR-205-3p) and microRNA-205-5p (miR-205-5p) on the cell proliferation, migration and invasion of head and neck squamous cell carcinoma (HNSCC). Methods The miRNA expression data were obtained from The Cancer Genome Atlas, and were used to determine miR-205 expression in HNSCC and paracancerous tissues. Quantitative real-time fluorescent polymerase chain reaction (qRT-PCR) was used to detect the expressions of miR-205-3p and miR-205-5p in HNSCC cell lines 5-8F, 6-10B, CNE2, Tu686 and mucosal tissues of nasopharynx. MiR-205-3p inhibitor, miR-205-5p inhibitor and miR-NC were transfected into Tu686 cells, and qRT-PCR was employed to evaluate the silence efficiency. In the following study, the cells were divided into four groups: miR-205-3p inhibitor and control, miR-205-5p inhibitor and control. Then the cell counting kit (CCK-8) assay, Transwell migration and invasion assays were carried out to examine the proliferation, migration and invasion abilities of the cells in the four groups. Results Compared with paracancerous tissues, the higher expression of miR-205 in HNSCC tissues was observed [M (QR): 61 012 (51 448) vs. 28 579 (35 959), Z=-6.420, P<0.001)]. The expression levels of miR-205-3p in HNSCC cell lines 5-8F, 6-10B, CNE2, Tu686 and mucosal tissues of nasopharynx were 0.36±0.07, 0.20±0.06, 0.15±0.04, 0.25±0.04 and 1.00±0.00 respectively, with a significant difference (F=162.71, P<0.001). The expression levels of miR-205-5p in cell lines 5-8F, 6-10B, CNE2, Tu686 and mucosal tissues of nasopharynx were 0.20±0.01, 0.21±0.01, 1.06±0.18, 23.61±2.07 and 1.00±0.00 respectively, with a significant difference (F=371.81, P<0.001). Compared with 5-8F, 6-10B, CNE2 cells, the expression of miR-205-3p in Tu686 cells was higher than those in 6-10B, CNE2 cells (P=0.195; P=0.020), and lower than that in 5-8F cells (P=0.023), and the expression of miR-205-5p in Tu686 cells was the highest (P<0.001; P<0.001; P<0.001). The expressions of miR-205-3p and miR-205-5p in Tu686 cells were effectively downregulated by inhibitors, and the silence efficiencies were 87% and 83%, respectively. The absorbance (A) values of miR-205-3p inhibitor group on the first, second, third, fourth and fifth day were 0.26±0.06, 0.55±0.11, 1.52±0.13, 1.91±0.07, 2.14±0.24, and those of miR-NC group were 0.29±0.07, 0.78±0.11, 1.59±0.15, 1.95±0.08, 2.02±0.12. There were no significant differences between the two groups (t=0.506, P=0.639; t=2.459, P=0.070; t=0.573, P=0.597; t=0.655, P=0.548; t=-0.759, P=0.490). The cell proliferations of miR-205-5p inhibitor group on the first, second, third, fourth and fifth day were 0.30±0.08, 0.61±0.08, 0.85±0.08, 1.08±0.12, 1.16±0.18, and those of miR-NC group were 0.41±0.10, 0.78±0.14, 1.33±0.28, 1.87±0.09, 2.08±0.19. The cell proliferations of miR-205-5p inhibitor group on the third, fourth and fifth day were significantly lower than those of miR-NC group (t=3.665, P=0.017; t=12.223, P<0.001; t=7.825, P<0.001). Transwell migration assay demonstrated that downregulation of miR-205-3p had no significant effect on migration abilities of Tu686 cells. The numbers of cell permeating septum of miR-205-3p inhibitor group and miR-NC group were 192.00±28.49 and 188.40±22.52, respectively. There was no significant difference between the two groups (t=-0.160, P=0.877). Downregulation of miR-205-5p significantly decreased the migration abilities of Tu686 cells. The numbers of cell permeating septum of miR-205-5p inhibitor group and miR-NC group were 109.40±27.63 and 183.60±31.63, respectively, and the difference was statistically significant (t=3.951, P=0.004). Transwell invasion assay showed that the numbers of cell permeating septum of miR-205-3p inhibitor group and miR-NC group were 93.40±10.24 and 96.20±16.56, respectively. There was no significant difference between the two groups (t=0.322, P=0.756). The numbers of cell permeating septum of miR-205-5p inhibitor group and miR-NC group were 53.00±17.80 and 94.40±14.38, respectively, and the difference was statistically significant (t=4.045, P=0.004). Conclusion Downregulation of miR-205-5p can inhibit the proliferation, migration and invasion abilities of Tu686 cells. However, downregulation of miR-205-3p has no significant effect on the proliferation, migration and invasion of Tu686 cells. Key words: Head and neck neoplasms; MicroRNAs; Cell proliferation; Cell movement; Neoplasm invasiveness

Silence were now a sin

Silence were now a sin

The regulation mechanism of silencing the DUSP1 gene on the release of proinflammatory cytokines in mice with acute pancreatitis

Objective To investigate the regulation mechanism of silencing the DUSP1 gene on the release of proinflammatory cytokines in mice with acute pancreatitis(AP). Methods Two DUSP1-siRNA and one scramble siRNA sequences were designed, and the sequence with higher silence efficiency was selected.Mice models with AP were established, and KM mice were divided into 6 groups: control group, AP group, AP+ PD98059 group, AP+ scramble group, AP+ siRNA group and AP+ PD98059+ siRNA group.Expressions of proinflammatory cytokines tumor necrosis factor-α(TNF-α), interleukin(IL)-1β and IL-6 in serum were detected by using enzyme linked immunosorbent assay(ELISA) after 12 h, 24 h, 48 h of modeling.Serum amylase levels were detected.The mRNA expression levels of DUSP1, TNF-α, IL-1β and IL-6 in pancreatic tissues were detected by using quantitative real time polymerase chain reaction (qPCR). The protein expression levels of DUSP1, extracellular regulated protein kinases(ERK), c-Jun N-terminal kinase(JNK), p38, p-ERK, p-JNK and p-p38 in pancreatic tissues were detected by using Western blot. Results Compared with the control group, the other 5 groups displayed the increased expressions of TNF-α, IL-1β, IL-6 and amylase in serum, and expressions of DUSP1, TNF-α, IL-1β, IL-6, p-ERK, p-JNK, p-p38 in tissues, and there was a statistical significance (all P<0.05). Compared with the AP group, the AP+ PD98059+ siRNA group showed the decreased DUSP1 expression in tissues, and there was a statistical significance (all P<0.05); the AP+ PD98059 group had decreased expressions of TNF-α, IL-1β, IL-6 and amylase in serum, and expressions of TNF-α, IL-1β, IL-6, p-ERK, p-JNK, p-p38 in tissues, and there were statistical significances (all P<0.05); while the opposite results were observed in the AP+ siRNA group with DUSP1 expression decreased. Conclusions The results support that silencing the DUSP1 gene promotes the release of proinflammatory cytokines through activating the mitogen-activated protein kinase signaling pathway in mice with AP. Key words: Mitogen-activated protein kinase signaling pathway; Acute pancreatitis; Proinflammatory cytokines; Lentiviral vectors; DUSP1 gene

"Minha história não é minha história. Ela é a história de uma geração" - A trajetória de uma militante dos movimentos sociais

Apesar da abertura do campo historiográfico para novas temáticas e novas abordagens, construir uma história das mulheres ainda é um desafio e emerge como um campo marcado por silêncios e lacunas. Nesse sentido, podemos compreender a história oral como uma estratégia para acessar a memória de agentes históricos silenciados e negligenciados. O presente artigo tem como objetivo resgatar elementos referentes à importantes movimentos sociais ocorridos no Brasil a partir da década de 1960, utilizando como fio condutor os relatos de militância de Ana Maria Gomes, socióloga, docente da Universidade Federal de Mato Grosso do Sul, integrante do denominado Grupo de Osasco e de movimentos de resistência à ditadura militar e reconhecida pela sua expressiva atuação no movimento de mulheres.História das mulheres; movimentos sociais; militância

Speaking with Silence

Speaking with Silence

Silence and Familial Homophobia in Colm Tóibín’s “Entiendes” and “One Minus One”

The present study focuses on two of Colm Toibin’s gay short-stories – “Entiendes” (1993) and “One Minus One” (2010) – in which the homosexual son meditates on his attachment to the dead mother. In both texts, Toibin characterises the mother-son bond as being fraught with silence, resentment and lack of communication. In “One Minus One” and “Entiendes”, the son’s closeted homosexuality coexists with familial legacies of shame, uneasiness and duplicity. The central characters in the two texts are similar, as they experience the same type of existential exile, solitude and alienation derived from their complex attachments to home and family. As shall be explained, the author dwells on the damaging effects of familial homophobia, highlighting the limitations of the dominant heteronormative family model to accommodate gay sensibilities.

Community radio news coverage in the European media : mapping popular issues and important silences

Una de les maneres adequades d’entendre la imatge social de les institucions, les personalitats i els temes mes importants per a una societat exigeix l’observacio del discurs mediatic (Macedo i Cabecinhas, 2014; Carvalho, 2010). Aquest article busca identificar els continguts que s’han publicat en els mitjans periodistics sobre el tema de les radios comunitaries per avaluar la presencia d’aquest tema en el discurs periodistic. No obstant aixo, les radios comunitaries viuen en un ambient totalment paradoxal a Europa, amb diferents percepcions sobre la utilitat que tenen aquests mitjans de comunicacio, aixi com les formes de regulacio legal d’aquests projectes en el mateix marc territorial (Carpentier i Dahlgren, 2011). Aquest treball analitza la projeccio mediatica de les radios comunitaries en el discurs periodistic avaluant les tematiques mes comunes, com tambe les que queden silenciades. A partir d’una analisi textual amb mes de cent articles periodistics de diferents mitjans de comunicacio generalistes europeus, aquest article conclou que les radios comunitaries son un tema que apareix mes als paisos on aquests projectes estan assenyalats i permesos per la llei. S’arriba a la conclusio que els exemples tractats en les noticies pertanyen al continent europeu i no s’incideix tant en els aspectes socials o legals de la repercussio social d’aquests mitjans.

The vitamin D hormone and its nuclear receptor: molecular actions and disease states.

Vitamin D plays a major role in bone mineral homeostasis by promoting the transport of calcium and phosphate to ensure that the blood levels of these ions are sufficient for the normal mineralization of type I collagen matrix in the skeleton. In contrast to classic vitamin D-deficiency rickets, a number of vitamin D-resistant rachitic syndromes are caused by acquired and hereditary defects in the metabolic activation of the vitamin to its hormonal form, 1,25-dihydroxyvitamin D3 (1,25(OH)2D3), or in the subsequent functions of the hormone in target cells. The actions of 1,25(OH)2D3 are mediated by the nuclear vitamin D receptor (VDR), a phosphoprotein which binds the hormone with-high affinity and regulates the expression of genes via zinc finger-mediated DNA binding and protein-protein interactions. In hereditary hypocalcemic vitamin D-resistant rickets (HVDRR), natural mutations in human VDR that confer patients with tissue insensitivity to 1,25(OH)2D3 are particularly instructive in revealing VDR structure function relationships. These mutations fall into three categories: (i) DNA binding/nuclear localization, (ii) hormone binding and (iii) heterodimerization with retinoid X receptors (RXRs). That all three classes of VDR mutations generate the HVDRR phenotype is consistent with a basic model of the active receptor as a DNA-bound, 1,25(OH)2D3-liganded heterodimer of VDR and RXR. Vitamin D responsive elements (VDREs) consisting of direct hexanucleotide repeats with a spacer of three nucleotides have been identified in the promoter regions of positively controlled genes expressed in bone, such as osteocalcin, osteopontin, beta 3-integrin and vitamin D 24-OHase. The 1,25(OH)2D3 ligand promotes VDR-RXR heterodimerization and specific, high affinity VDRE binding, whereas the ligand for RXR, 9-cis retinoic acid (9-cis RA), is capable of suppressing 1,25(OH)2D3-stimulated transcription by diverting RXR to form homodimers. However, initial 1,25(OH)2D3 liganding of a VDR monomer renders it competent not only to recruit RXR into a heterodimer but also to conformationally silence the ability of its RXR partner to bind 9-cis RA and dissociate the heterodimer. Additional probing of protein-protein interactions has revealed that VDR also binds to basal transcription factor IIB (TFIIB) and, in the presence of 1,25(OH)2D3, an RXR-VDR-TFIIB ternary complex can be created in solution. Moreover, for transcriptional activation by 1,25(OH)2D3, both VDR and RXR require an intact short amphipathic alpha-helix, known as AF-2, positioned at their extreme C-termini. Because the AF-2 domains participate neither in VDR-RXR heterodimerization nor in TFIIB association, it is hypothesized that they contact, in a ligand-dependent fashion, transcriptional coactivators such as those of the steroid receptor coactivator family, constituting yet a third protein-protein interaction for VDR. Therefore, in VDR-mediated transcriptional activation, 1,25(OH)2D3 binding to VDR alters the conformation of the ligand binding domain such that it: (i) engages in strong heterodimerization with RXR to facilitate VDRE binding, (ii) influences the RXR ligand binding domain such that it is resistant to the binding of 9-cis RA but active in recruiting coactivator to its AF-2 and (iii) presents the AF-2 region in VDR for coactivator association. The above events, including bridging by coactivators to the TATA binding protein and associated factors, may position VDR such that it is able to attract TFIIB and the balance of the RNA polymerase II transcription machinery, culminating in repeated transcriptional initiation of VDRE-containing, vitamin D target genes. Such a model would explain the action of 1,25(OH)2D3 to elicit bone remodeling by stimulating osteoblast and osteoclast precursor gene expression, while concomitantly triggering the termination of its hormonal signal by inducing the 24-OHase catabolizing enzyme.

Effects of short hairpin RNA-mediated hypoxia inducible factor-1α gene silence on the proliferation of osteosarcoma cells under hypoxic condition

Objective To explore the effect of hypoxic condition on the proliferation of primary osteosarcoma cells, and the proliferation change of primary osteosarcoma cells under hypoxia after hypoxia inducible factor (HIF)-1α gene silence with short hairpin RNA (shRNA). Methods Primary osteosarcoma cells were isolated from osteosarcoma tumor and cultured under hypoxic (37℃, 1% O2, 5% CO2) or normoxic (37℃, 21% O2, 5% CO2) condition separately. Firstly, the cell proliferation, cell cycle and the HIF-1α protein expression of the cells were detected by methyl thiazol tetrazolium (MTT) assay, flow cytometry and Western blotting separately. Secondly, the primary osteosarcoma cells were transfected by lentiviral vectors carrying HIF-1α or scramble (SCR) shRNA, which was respectively denoted as HIF-1α/shRNA group or SCR/shRNA group. HIF-1α mRNA expression in both groups was quantified using quantitative polymerase chain reaction (qPCR) and the HIF-1α protein expression was detected by Western blotting. Finally, the transfected cells were cultured under hypoxic condition, and subsequently MTT assay was carried out for the evaluation of their proliferation. Results Hypoxia could inhibit the proliferation of primary osteosarcoma cells within 24 h, and this inhibition attenuated with the continually hypoxic condition. The expression of HIF-1α increased under continually hypoxic condition, indicating that the osteosarcoma cell proliferation might be related to the HIF-1α expression, and the cell proliferation rate of hypoxia cells decreased by 30.5% in 12 h and 25.0% in 24 h. As compared with SCR/shRNA group, the mRNA and protein expression levels of HIF-1α in HIF-1α/shRNA group significantly decreased (P=0.000), suggesting that the specific gene silence of HIF-1α could inhibit the osteosarcoma cell proliferation. Conclusion shRNA-mediated HIF-1α gene silence can effectively reverse the promoting effect of HIF-1α on osteosarcoma cell proliferation under hypoxic condition, which may further interfere with the osteosarcoma development and growth. Key words: Osteosarcoma; Hypoxia-inducible factor-1α; Gene silence; Short hairpin RNA

LA CONSTITUCIÓN SENTIMENTAL. PROSTITUCIÓN, TRABAJO SEXUAL Y TRATA DE PERSONAS EN COLOMBIA

El propósito central de este ensayo consiste en contextualizar el debate contemporáneo entre la abolición de la prostitución o la regulación el trabajo sexual en la dinámica de un humanitarismo sexual altamente sentimentalizado. El texto muestra, en primer lugar, que la compasión —como fuerza central que mueve al humanitarismo sexual a “rescatar” a las víctimas de explotación sexual— no es necesariamente, como sostienen algunos, una fuerza que silencia a quien la recibe. Las diferencias de contexto y de dinámicas políticas, pueden determinar que, en ciertas ocasiones, la compasión efectivamente silencie, mientras que en otras puede dar lugar a dinámicas sociales verdaderamente emancipatorias. El artículo propone, así, que, en cada caso concreto en que aparece la compasión como fuerza movilizadora, se haga un análisis pragmático de los costos y beneficios de este uso. En segundo lugar, el ensayo traslada el marco conceptual anterior a la discusión constitucional sobre la prostitución y el trabajo sexual en Colombia y muestra cómo ésta ocurre en un terreno dominado por un humanitarismo sexual compasivo que ha determinado la “sentimentalización” de los valores de dignidad, autonomía e igualdad. Un análisis pragmático de la jurisprudencia de la Corte Constitucional colombiana sobre prostitución, trabajo sexual y tráfico de personas permite concluir, al menos provisionalmente, que la señalada sentimentalización constitucional sustenta, de manera simultánea, el proyecto político de la regulación del trabajo sexual y el de la abolición de la prostitución.

Effects of resveratrol on cognitive dysfunction and the expression of parvalbumin in depression model mice

Objective To examine the antidepressant effects of resveratrol (RSV), and its possible mechanism specialized on improving cognitive function. Methods Thirty-two C57BL/6J mice were randomly divided into four groups: Control, Model, Model+ RSV and Model+ NA+ RSV groups.The mice were subjected corticosterone (20 mg/(kg·d)) intraperitoneal injection for 21 consecutive days except the control mice.From the 22nd to 42nd day, the mice in different groups received further treatment with vehicle/ RSV (400 mg/(kg·d), op)/ NA (100 mg/(kg·d), ip)+ RSV (400 mg/(kg·d), op) .The sugar preference test, novel object recognition test, novel location recognition test and water maze test were applied to evaluate the cognitive effects of RSV on mice.Subsequently, the silence information regulation factor 1(SIRT1), peroxisome proliferator-activated receptor-γ coactivator-1α(PGC-1α), parvalbumin(PV) transcription and translation level were evaluated by RT-PCR and Western blotting. Results The sugar preference test showed that the depression model mediated by CORT injection was successfully established(F(1, 30)=6, P=0.038). In in the novel object learning test, resveratrol significantly increased the proration on the frequency((-0.20±0.37) vs ( 0.16±0.29) )and duration((0.10±0.45) vs ( 0.62±0.29)) and decreased the proration on the distance((0.09±0.36) vs (0.55±0.27)). In the water maze test, resveratrol reduced the time((41±9 )s; (26±8)s) and distance ((295±70)cm; (224±43)cm)to find the platform.All the results were accompanied with the increased expression of protein SIRT1 (F(3, 29)=15.60, P<0.01), PGC-1α (F(3, 29)=7.51, P=0.0006) and PV (F(3, 29)=17.87, P=0.0004). While pretreatment with nicotinamide, resveratrol could not rescue the cognitive impairment and could not reverse the iecreased expression level of protein SIRT1, PGC-1α and PV. Conclusion Resveratrol can reverse the cognitive dysfunction of depressant mice, which may be achieved by activating the SIRT1/ PGC-1α signaling pathway and increasing the transcription and protein expression of PV. Key words: Resveratrol; Depression; Cognition; SIRT1; PGC-1α; Parvalbumin; Mice

The regulation of hypoxia inducible factors on glucose transporter 1, pyruvate kinase M2 and vascular endothelial growth factor in BGC-823 cells

Objective To compare the expression of glucose transporter 1 (GLUT1), pyruvate kinase M2 (PKM2), vascular endothelial growth factor (VEGF) in BGC-823 cells after transfected with small interfering RNA of hypoxia-inducible factors (HIF-1α, HIF-2α, HIF-1β) and investigate the effect of hypoxia inducible factors (HIFs) on the regulation of GLUT1, PKM2 and VEGF in gastric cancer. Methods Small RNA interference (RNAi) was used to silence BGC-823 cells. The mRNA and protein expression levels of HIFs, GLUT1, PKM2 and VEGF were detected by quantitative real-time reverse transcription polymerase chain reaction (PCR) and Western blotting. Results The mRNA (0.372±0.097) and protein (0.351±0.041) expression of HIF-1α was lower than in empty plasmid group (0.980±0.115 and 0.520±0.030) and blank control group (1.000±0.000 and 0.478±0.062) after infection with RNAi, the differences were all statistically significant (F=50.358, P=0.000; F=10.931, P=0.010). The mRNA (0.391±0.091) and protein (0.293±0.031) expression of HIF-2α was lower than in empty plasmid group (0.980±0.115 and 0.520±0.030) and blank control group (1.000±0.000 and 0.430±0.056) after infection with RNAi, the differences were all statistically significant (F=72.416, P=0.000; F=9.016, P=0.016). The mRNA (0.383±0.091) and protein (0.323±0.038) expression of HIF-1β was lower than in empty plasmid group (0.987±0.100 and 0.445±0.059) and blank control group (1.000±0.000 and 0.457±0.040) after infection with RNAi. The differences were all statistically significant (F=78.273, P=0.000; F=7.636, P=0.022). The mRNA (GLUT1: 0.721±0.067, 0.833±0.059, 0.493±0.067; VEGF: 0.711±0.057, 0.826±0.054, 0.793±0.068) and protein (GLUT1: 0.263±0.072, 0.303±0.042, 0.198±0.057; VEGF: 0.391±0.049, 0.451±0.054, 0.473±0.086) expression of GLUT1 and VEGF was all decreased after silencing HIFs (HIF-1α, HIF-1β, HIF-2α) when compared with empty plasmid group (mRNA: 1.046±0.094, 1.039±0.092; protein: 0.391±0.064, 0.690±0.066) and blank control group (mRNA: 1.000±0.000, 1.000±0.000; protein: 0.438±0.055, 0.707±0.040) and the differences were all statistically significant (VEGF mRNA: F=35.232, P=0.000; GLUT1 mRNA: F=15.364, P=0.000; VEGF protein: F=8.150, P=0.003, GLUT1 protein: F=17.109, P=0.000). The levels of VEGF decreased the most obvious when transfection with HIF-2α siRNA, while the levels of GLUT1 decreased the most obvious when transfection with HIF-1α siRNA. The mRNA (0.611±0.036) and protein (0.351±0.047) expression of PKM2 was all decreased after silencing HIF-1α when compared with empty plasmid group (1.037±0.197 and 0.731±0.057) and blank control group (1.000±0.000 and 0.737±0.043), the differences were all statistically significant, (F=12.480, P=0.007; F=60.403, P=0.000). Whlie only mRNA (0.793±0.067) decreased after silencing HIF-2α, the differences were statistically significant (F=12.480, P=0.007). The protein changed a little (0.675±0.030, F=1.211, P=0.373), and both mRNA (0.980±0.115) and protein (0.641±0.076) had no significant change after transfection with HIF-1β (F=0.144, P=0.869; F=2.404, P=0.171). Conclusion The results suggest that HIF-1α, HIF-2α and HIF-1β are all involved in regulating GLUT1 and VEGF. HIF-1α pathway plays a major role in regulating PKM2 and HIF-2α may regulate PKM2 in transcription level. Key words: Gastric cancer; Hypoxia-inducible factors; Pyruvate kinase M2; Glucose transporter 1; Vascular endothelial growth factor

Listen and the question of silence

Abstract Listen is a film about words, but around words. The words become useless and are surrounded by silence. And the whole film is constructed on this silence, which builds up like an unbreakable wall. The question is thus: what are we listening to? What should we listen to? And maybe, even more crucial: who should be listening?

Behind our silence: Refugees feel that they are not allowed to give their views in public in case they upset their new nation, they tell our interviewer

Behind our silence: Refugees feel that they are not allowed to give their views in public in case they upset their new nation, they tell our interviewer

The effects of targeted inhibition of hedgehog signaling pathway on proliferation, apoptosis and metastasis of hepatocellular carcinoma

Objective To observe Hedgehog signaling pathways of liver cancer cell growth and the influence of the metastatic potential targeted inhibit Hedgehog. Methods Construction of Smo shRNA plasmid, The stable and low-expressed Smo-expressing HCC QGY-7701 cell line was screened after lipofection. The stable and low-expressed Smo-expressing HCC QGY-7701 cell line was screened, The cell cycle, apoptosis, invasion and metastasis of QGY-7701 cells were detected by Western blot, flow cytometry, CCK8 and transwell assay. Subcutaneous implantation of hepatocarcinoma cells in nude mice. Study on the growth and metastasis of hepatocarcinoma cells with low expression of Smo in. The ultrastructural changes of hepatoma cells with low expression of Smo were observed under electron microscope. Results RT-PCR and Western blot showed stable shR-Smo cell line was successfully constructed. Cell cycle test showed that compared with the control group, G0/G1 cells increased in shR-Smo, cells in S phase decreased; apoptosis, CCK8 and Transwell tests showed that Smo-gene silencing could significantly increase the apoptosis percentage of the hepatic cancer cells to (5.46%±1.46%), proliferation activity decreasedand and the migration rates reduced to (7.82%±2.14)%; nude mice model showed that Smo-gene silencing could inhibit the growth of hepatocellular carcinoma cells in vivo, electron microscopy revealed that lysosomes increased significantly in Smo-gene silence cells. Conclusions Blocking Hh signaling pathways , liver cancer cells in vitro malignant degree of decline. Hedgehog in treating liver cancer have hidden meaning. Key words: Hepatocelluar Carcinoma; Signal transduction; Hedgehog signaling pathway