Acidic Environment Induces Pyroptosis of NLRP3 Inflammatory Vesicle-Mediated Nucleus Pulposus Cells Through Up-Regulation of POSTN Expression

SUMMARY: Intervertebral disc degeneration (IVDD) is induced by nucleus pulposus (NP) dysfunction as a result of massive loss of NP cells. It has been reported that the acidic microenvironment of the intervertebral disc (IVD) can induce NP cell pyroptosis, and that up-regulation of periostin (POSTN) expression has a negative effect on NP cell survival. However, the relationship between the acidic environment, POSTN expression level and NP cell pyroptosis is unclear. Therefore, the aim of this study was to explore the relationship between acidic environment and POSTN expression level in NP cells, as well as the effect of POSTN in acidic environment on NP cell pyroptosis. NP cells were obtained from the lumbar vertebrae of Sprague Dawley (SD) male rats. These cells were divided into normal and acidic groups according to whether they were exposed to 6 mM lactic acid solution. And NP cells in the acidic group were additionally divided into three groups: (1) Blank group: no transfection; (2) NC group: cells transfected with empty vector plasmid; (3) sh-POSTN group: cells transfected with sh-POSTN plasmid to knock down the expression level of POSTN. Quantitative real-time PCR (qRT-PCR) and western blot was performed to assess the expression of POSTN at the mRNAand protein levels. CCK8 was used to evaluate cell survival. Western blot, in addition, was performed to examine acid-sensing ion channels (ASIC)-related proteins. And pyroptosis was detected by ELISA and western blot. The expression level of POSTN was significantly increased in NP cells in acidic environment. Knockdown of POSTN expression promoted the survival of NP cells in acidic environment and reduced the protein levels of ASIC3 and ASIC1a in NP cells. Moreover, knockdown of POSTN expression decreased the pyroptosis proportion of NP cells and the levels of pro-inflammatory cytokines interleukin (IL)-1β and IL-18. The levels of pyroptosis-related proteins NLRP3, ASC, cleaved-Caspase-1, and cleaved-GSDMD were also affected by the decreased POSTN expression. The extracellular acidic environment created by lactic acid solution activated NLRP3 inflammatory vesicle-induced caspase-1 to get involved in NP cell pyroptosis by up-regulating POSTN expression.

La degeneración del disco intervertebral (DDIV) es inducida por una disfunción del núcleo pulposo (NP) como resultado de una pérdida masiva de células NP. Se ha informado que el microambiente ácido del disco intervertebral (DIV) puede inducir la piroptosis de las células NP y que la regulación positiva de la expresión de periostina (POSTN) tiene un efecto negativo en la supervivencia de las células NP. Sin embargo, la relación entre el ambiente ácido, el nivel de expresión de POSTN y la piroptosis de las células NP es poco clara. Por lo tanto, el objetivo de este estudio fue explorar la relación entre el ambiente ácido y el nivel de expresión de POSTN en células NP, así como el efecto de POSTN en ambiente ácido sobre la piroptosis de las células NP. Las células NP se obtuvieron de las vertebras lumbares de ratas macho Sprague Dawley (SD). Estas células se dividieron en grupos normales y ácidos según se expusieron a una solución de ácido láctico 6 mM. Las células NP en el grupo ácido se dividieron adicionalmente en tres grupos: (1) Grupo en blanco: sin transfección; (2) grupo NC: células transfectadas con plásmido vector vacío; (3) grupo sh-POSTN: células transfectadas con plásmido sh-POSTN para reducir el nivel de expresión de POSTN. Se realizó una PCR cuantitativa en tiempo real (qRT-PCR) y una transferencia Western para evaluar la expresión de POSTN en los niveles de ARNm y proteína. Se utilizó CCK8 para evaluar la supervivencia celular. Además, se realizó una transferencia Western para examinar las proteínas relacionadas con los canales iónicos sensibles al ácido (ASIC). La piroptosis se detectó mediante ELISA y Western blot. El nivel de expresión de POSTN aumentó significativamente en células NP en ambiente ácido. La eliminación de la expresión de POSTN promovió la supervivencia de las células NP en un ambiente ácido y redujo los niveles de proteína de ASIC3 y ASIC1a en las células NP. Además, la eliminación de la expresión de POSTN disminuyó la proporción de piroptosis de las células NP y los niveles de citocinas proinflamatorias interleucina (IL) - 1β e IL-18. Los niveles de proteínas relacionadas con la piroptosis NLRP3, ASC, Caspasa-1 escindida y GSDMD escindida también se vieron afectados por la disminución de la expresión de POSTN. El ambiente ácido extracelular creado por la solución de ácido láctico activó la caspasa-1 inducida por vesículas inflamatorias NLRP3 para involucrarse en la piroptosis de las células NP mediante la regulación positiva de la expresión de POSTN.

Detection of DNA methylation of HYAL2 gene for differentiating malignant from benign thyroid tumors / 南方医科大学学报

OBJECTIVE@#To assess the value of DNA methylation level of HYAL2 gene as a molecular marker for differential diagnosis of malignant and benign thyroid tumors.@*METHODS@#DNA methylation of HYAL2 gene in tissue specimens of 190 patients with papillary thyroid cancer (PTC) and 190 age- and gender-matched patients with benign thyroid tumors was examined by mass spectrometry, and the protein expression of HYAL2 was detected immunohistochemically for another 55 pairs of patients. Logistic regression analysis was performed to calculate the odds ratio (OR) and evaluate the correlation of per 10% reduction in DNA methylation with PTC. Receiver operating characteristic (ROC) curve analysis was performed and the area under curve (AUC) was calculated to assess the predictive value of alterations in HYAL2 methylation.@*RESULTS@#Hypomethylation of HYAL2_CpG_3 was significantly correlated with early-stage PTC (OR=1.51, P=0.001), even in stage I cancer (OR=1.42, P=0.007). Age-stratified analysis revealed a significantly stronger correlation between increased HYAL2_CpG_ 3 methylation and early-stage PTC in patients below 50 years than in those older than 50 years (OR: 1.89 vs 1.37, P < 0.05); ROC analysis also showed a larger AUC of 0.787 in younger patients. The results of immunohistochemistry showed that patients with PTC had significantly higher protein expressions of HYAL2 than patients with benign tumors.@*CONCLUSION@#The alterations of DNA methylation level of HYAL2 gene is significantly correlated with early-stage PTC, suggesting the value of DNA methylation level as a potential biomarker for differentiation of malignant from benign thyroid tumors.

Aqueous humor heat-shock protein 70, periostin, and irisin levels in patients with pseudoexfoliation syndrome / Níveis de proteína de choque térmico 70, de periostina e de irisina no humor aquoso em pacientes com pseudoexfoliação

ABSTRACT Purpose: To measure humor heat-shock protein 70, periostin, and irisin levels in patients with pseudoexfoliation syndrome and cataract (without glaucoma), and compare them with those of patients with cataract but without pseudoexfoliation. Methods: We examined 31 eyes of 31 patients with pseudoexfoliation and cataract (without glaucoma) and 30 eyes of 30 patients with cataract. We collected aqueous humor samples from all patients at the time of cataract surgery through a limbal paracentesis via a 25-gauge cannula mounted on a tuberculin syringe that received 100 to 150 µL of aqueous humor. We measured levels of aqueous humor Heat shock protein 70, periostin, and irisin using enzyme-linked immunosorbent assay methods. Results: The age (p=0.221) and gender (p=0.530) means were similar between the pseudoexfoliation and control groups. The mean Heat shock protein 70 level (29.22 ± 9.46 ng/mL; 17.88-74.46) in the pseudoexfoliation group was significantly higher than that in the control group (19.03 ± 7.05 ng/mL; 9.93-35.52; p<0.0001). The mean periostin level was significantly higher (6017.32 ± 1271.79 pg/mL; 3787.50-10803.57) in the pseu doexfoliation group than that in the control group (4073.63 ± 1422.79 pg/mL; 2110.44-7490.64; p<0.0001). The mean irisin level (53.77 ± 10.19 ng/mL; 29.46-71.16) was significantly higher than that in the control group (39.29 ± 13.58 ng/mL; 19.41-70.56; p<0.0001). Conclusions: Heat shock protein 70, periostin, and irisin levels increase in the aqueous humor of patients with pseudoexfoliation without glaucoma.

RESUMO Objetivo: Comparar os níveis de proteína de choque térmico 70, de periostina e de irisina no humor aquoso de pacientes com pseudoexfoliação com catarata sem glaucoma e compará-los com pacientes com catarata sem pseudoexfoliação. Métodos: Trinta e um olhos de 31 pacientes com pseudoexfoliação com catarata sem glaucoma e 30 olhos de 30 indivíduos com catarata foram incluídos neste estudo. Amostras de humor aquoso foram coletadas de todos os pacientes no momento da cirurgia de catarata e obtidas através de uma paracentese límbica por meio de uma cânula de calibre 25 acoplada a uma seringa com tuberculina. Foram coletados 100 a 150 µL de humor aquoso. Os níveis de proteína de choque térmico 70, de periostina e de irisina no humor aquoso foram medidos usando o método de ensaio imunossorvente ligado a enzima. Resultados: A média da idade (p=0,221) e sexo (p=0,530) foram semelhantes entre os grupos pseudoexfoliação e controle. Os níveis médios de proteína de choque térmico 70 foram 29,22 ± 9,46 ng/mL (17,88-74,46) e 19,03 ± 7,05 ng/ mL (9,93-35,52) nos grupos pseudoexfoliação e controle, respectivamente. Os níveis de proteína de choque térmico 70 foram maiores no grupo pseudoexfoliação (p<0,0001). O nível médio de periostina foi de 6017,32 ± 1271,79 pg/mL (3787,50-10803,57) no grupo pseudoexfoliação e 4073,63 ± 1422,79 pg/mL (2110,44-7490,64) no grupo controle. O nível médio de periostina também foi maior no grupo pseudoexfoliação (p<0,0001). Os níveis médios de irisina foram 53,77 ± 10,19 ng/mL (29,46-71,16) e 39,29 ± 13,58 ng/mL (19,41-70,56) nos grupos pseudoexfoliação e controle, respectivamente. O nível médio de irisina foi maior no grupo pseudoexfoliação do que no grupo controle (p<0,0001). Conclusões: Os níveis de proteína de choque térmico 70, de periostina e de irisina aumentam no humor aquoso de pacientes com pseudoexfoliação sem glaucoma.

MiR-128 suppresses metastatic capacity by targeting metadherin in breast cancer cells

BACKGROUND: Breast cancer, the most common cancer in women worldwide, causes the vast majority of cancer-related deaths. Undoubtedly, tumor metastasis and recurrence are responsible for more than 90 percent of these deaths. MicroRNAs are endogenous noncoding RNAs that have been integrated into almost all the physiological and pathological processes, including metastasis. In the present study, the role of miR-128 in breast cancer was investigated. RESULTS: Compared to the corresponding adjacent normal tissue, the expression of miR-128 was significantly suppressed in human breast cancer specimens. More importantly, its expression level was reversely correlated to histological grade of the cancer. Ectopic expression of miR-128 in the aggressive breast cancer cell line MDA-MB-231 could inhibit cell motility and invasive capacity remarkably. Afterwards, Metadherin (MTDH), also known as AEG-1 (Astrocyte Elevated Gene 1) and Lyric that implicated in various aspects of cancer progression and metastasis, was further identified as a direct target gene of miR-128 and its expression level was up-regulated in clinical samples as expected. Moreover, knockdown of MTDH in MDA-MB-231 cells obviously impaired the migration and invasion capabilities, whereas re-expression of MTDH abrogated the suppressive effect caused by miR-128. CONCLUSIONS: Overall, these findings demonstrate that miR-128 could serve as a novel biomarker for breast cancer metastasis and a potent target for treatment in the future.

Hippocampal overexpression of Down syndrome cell adhesion molecule in amyloid precursor protein transgenic mice

Down syndrome cell adhesion molecule (DSCAM) is located within the Down syndrome critical region of chromosome 21. DSCAM is a broadly expressed neurodevelopmental protein involved in synaptogenesis, neurite outgrowth, and axon guidance. We previously demonstrated DSCAM overexpression in the cortex of amyloid precursor protein (APP) transgenic mice, suggesting possible regulatory interactions between APP and DSCAM. APP mice exhibit deficits in hippocampus-dependent learning and memory. In this preliminary study, we examined age-related changes in DSCAM expression within the hippocampus in 16 APP transgenic mice (1, 3, 6 and 12 months old). Hippocampus-dependent spatial memory was assessed in APP mice and age-matched wild type littermates (WTs) using the Morris water maze (MWM). The cellular distribution of hippocampal DSCAM and total expression at both mRNA and protein levels were measured by immunohistochemistry, qRT-PCR, and western blotting, respectively. APP mice exhibited spatial memory deficits in the MWM. Intense DSCAM immunoreactivity was observed in the dentate gyrus granule cell layer and hippocampal stratum pyramidale. Total hippocampal DSCAM mRNA and protein expression levels were substantially higher in APP mice than WTs at 1 and 3 months of age. Expression decreased with age in both groups but remained higher in APP mice. DSCAM is overexpressed in the hippocampus over the first 12 months of life in APP mice, but especially during maturation to adulthood. In conclusion, these results suggest an association between DSCAM and APP mice, which is characterized by neuropathology and behavioral deficits. These results provide some clues for future studies on the role of DSCAM overexpression in the precocious cognitive decline observed in APP transgenic mice.

Circulatory and Ventilatory Power: Characterization in Patients with Coronary Artery Disease / Potência Circulatória e Ventilatória: Caracterização em Pacientes com Doença Arterial Coronariana

Background: Circulatory power (CP) and ventilatory power (VP) are indices that have been used for the clinical evaluation of patients with heart failure; however, no study has evaluated these indices in patients with coronary artery disease (CAD) without heart failure. Objective: To characterize both indices in patients with CAD compared with healthy controls. Methods: Eighty-seven men [CAD group = 42 subjects and healthy control group (CG) = 45 subjects] aged 40–65 years were included. Cardiopulmonary exercise testing was performed on a treadmill and the following parameters were measured: 1) peak oxygen consumption (VO2), 2) peak heart rate (HR), 3) peak blood pressure (BP), 4) peak rate-pressure product (peak systolic HR x peak BP), 5) peak oxygen pulse (peak VO2/peak HR), 6) oxygen uptake efficiency (OUES), 7) carbon dioxide production efficiency (minute ventilation/carbon dioxide production slope), 8) CP (peak VO2 x peak systolic BP) and 9) VP (peak systolic BP/carbon dioxide production efficiency). Results: The CAD group had significantly lower values for peak VO2 (p < 0.001), peak HR (p < 0.001), peak systolic BP (p < 0.001), peak rate-pressure product (p < 0.001), peak oxygen pulse (p = 0.008), OUES (p < 0.001), CP (p < 0.001), and VP (p < 0.001) and significantly higher values for peak diastolic BP (p = 0.004) and carbon dioxide production efficiency (p < 0.001) compared with CG. Stepwise regression analysis showed that CP was influenced by group (R2 = 0.44, p < 0.001) and VP was influenced by both group and number of vessels with stenosis after treatment (interaction effects: R2 = 0.46, p < 0.001). Conclusion: The indices CP and VP were lower in men with CAD than healthy controls. .

Fundamento: Os índices da Potência Circulatória (PC) e Potência Ventilatória (PV) têm sido utilizados para avaliação clínica de pacientes com insuficiência cardíaca, mas nenhum estudo avaliou esses índices em pacientes com Doença Arterial Coronariana (DAC). Objetivo: Caracterizar ambos os índices em pacientes com DAC comparados a indivíduos saudáveis. Métodos: Oitenta e sete homens [grupo DAC = 42 sujeitos e, grupo controle (GC) = 45 sujeitos] com idade entre 45 e 65 anos foram incluídos. Um Teste de Exercício Cardiopulmonar (TECP) foi realizado em esteira e as seguintes variáveis foram obtidas: 1) consumo de oxigênio (VO2) pico; 2) Frequência Cardíaca (FC) pico; 3) Pressão Arterial (PA) pico; 4) duplo produto pico (PA sistólica pico x FC pico); 5) pulso de oxigênio pico (VO2 pico dividido pela FC pico); 6) eficiência ventilatória para o consumo de oxigênio (OUES); 7) eficiência ventilatória para a produção de dióxido de carbono (VE/VCO2 slope); 8) PC (VO2 pico x PA sistólica pico); e 9) PV (PA sistólica pico dividido pelo VE/VCO2 slope). Resultados: O grupo DAC apresentou valores significativamente menores das seguintes variáveis no pico do exercício: VO2 (p < 0,001), FC (p < 0,001), PA sistólica (p < 0,001), duplo produto (p < 0,001), pulso de oxigênio (p = 0,008), OUES (p < 0,001), PC (p < 0,001) e PV (p < 0,001), e valores significativamente maiores de PA diastólica (p = 0,004) e VE/VCO2 slope (p < 0,001) em relação ao GC. Uma análise de regressão pelo método stepwise mostrou que a PC foi influenciada pelo grupo (R2 = 0,44, p < 0,001) e a PV tanto pelo grupo quanto pelo número de vasos com estenose pós tratamento (efeito de interação: R2 = 0,46, p < 0,001). Conclusion: Os índices da PC e PV foram menores em homens com DAC comparados ao GC, podendo dessa forma ser utilizados na caracterização dessa população. .

Adhesion molecules in breast carcinoma: a challenge to the pathologist / Moléculas de adesão no carcinoma mamário: um desafio ao patologista

The role of adhesion molecules is very important both in the activation of carcinogenesis and in the differentiation of subtypes of breast carcinoma, aiding in diagnosis, prognosis and therapeutic choice in these tumors. Therefore, understanding the functions and interrelationships among these molecules is crucial to the pathologist, who often uses these factors as a resource to differentiate tumors and further classify them according to a molecular point of view. Our goal is to describe the applicability and the difficulties encountered by the pathologist in the diagnosis of breast carcinoma, discussing the most commonly used markers of adhesion in routine analyses.

O papel das moléculas de adesão é de suma importância tanto na ativação da carcinogênese quanto na diferenciação dos subtipos de carcinomas mamários, auxiliando no diagnóstico, no prognóstico e na escolha terapêutica nessas neoplasias. Portanto, a compreensão das funções e das inter-relações entre essas moléculas é de suma importância para o patologista, que, muitas vezes, as utiliza como recurso na diferenciação dos tumores e, consequentemente, elas auxiliam em uma posterior classificação do ponto de vista molecular. O objetivo é descrever a aplicabilidade e as dificuldades encontradas pelo médico patologista no diagnóstico de carcinoma mamário, discutindo os marcadores de adesividade mais utilizados na rotina.

The Effect of Bortezomib on Expression of Inflammatory Cytokines and Survival in a Murine Sepsis Model Induced by Cecal Ligation and Puncture

PURPOSE: Although the proteasome inhibitor known as bortezomib can modulate the inflammatory process through the nuclear factor-kappa B signaling pathway, the immunomodulatory effect of pre-incubated bortezomib has not been fully evaluated for inflammation by infectious agents. Therefore, we evaluated the effect of bortezomib on the expression of inflammatory cytokines and mediators in macrophage cell lines and on survival in a murine peritonitis sepsis model. MATERIALS AND METHODS: Bortezomib was applied 1 hr before lipopolysaccharide (LPS) stimulation in RAW 264.7 cells. The cecal ligation and puncture (CLP) experiments were performed in C57BL/6J mice. RESULTS: Pre-incubation with bortezomib (25 nM or 50 nM) prior to LPS (50 ng/mL or 100 ng/mL) stimulation significantly recovered the number of viable RAW 264.7 cells compared to those samples without pre-incubation. Bortezomib decreased various inflammatory cytokines as well as nitric oxide production in LPS-stimulated cells. The 7-day survival rate in mice that had received bortezomib at 0.01 mg/kg concentration 1 hr prior to CLP was significantly higher than in the mice that had only received a normal saline solution of 1 mL 1 hr prior to CLP. In addition, the administration of bortezomib at 0.01 mg/kg concentration 1 hr before CLP resulted in a significant decrease in inflammation of the lung parenchyma. Collectively, pretreatment with bortezomib showed an increase in the survival rate and changes in the levels of inflammatory mediators. CONCLUSION: These results support the possibility of pretreatment with bortezomib as a new therapeutic target for the treatment of overwhelming inflammation, which is a characteristic of severe sepsis.

Role of acute alcohol poisoning and craniocerebral trauma in the mechanism of death caused by subarachnoid hemorrhage / 法医学杂志

OBJECTIVE@#To investigate the relation between the expression of tPA, MMP-2, MMP-9 and AEG-1 in the human brain tissue and the ethanol concentration under the acute alcohol poison, and to analyze the role of alcohol and trauma in the mechanism of death of subarachnoid hemorrhage.@*METHODS@#Fifteen real cases were collected in this study. The brain tissues were researched by histological examination and the concentration of ethanol in heart blood were detected. The tPA, MMP-2, MMP-9 and AEG-1 in brainstem, brain and cerebellum were observed respectively by immunohistochemistry.@*RESULTS@#In alcohol poisoning groups with or without trauma, the acute alcohol toxicity resulted in the swelling of brain tissues. The tPA, MMP-2, MMP-9 and AEG-1 of brainstem, brain and cerebellum showed high expression in alcohol victims, and the tPA in cerebellum showed no difference. The expression of the MMP-2, MMP-9 and AEG-1 showed good relation with the ethanol concentration in blood (P < 0.05, r > 0.6).@*CONCLUSION@#The expressions of tPA, MMP-2, MMP-9 and AEG-1 are significant higher in alcohol victims, and expressions of MMP-2 and MMP-9 and AEG-1 have positive correlation with the alcohol concentration. The alcohol has acute toxicity to brain cells.

Prognostic impact of epithelial cell adhesion molecule in ovarian cancer patients / 부인종양

No abstract available.

Overexpression of the epithelial cell adhesion molecule is associated with a more favorable prognosis and response to platinum-based chemotherapy in ovarian cancer / 부인종양

OBJECTIVE: Epithelial cell adhesion molecule (EpCAM) has experienced a renaissance lately as a binding site for targeted therapy as well as a prognostic marker in epithelial malignancies. Aim of this study was to study EpCAM as a potential prognostic marker in epithelial ovarian cancer (EOC). METHODS: EpCAM expression was assessed by immunohistochemistry on paraffin-embedded primary EOC-tissue samples. EpCAM overexpression was defined as an expression of EpCAM of 76% to 100%. Tissue samples and clinical data were systematically collected within the international and multicenter "Tumorbank Ovarian Cancer" network. RESULTS: Seventy-four patients, diagnosed with EOC between 1994 and 2009, were included in the study (median age, 56 years; range, 31 to 86 years). The majority of the patients (81.1%) presented with an advanced stage International Federation of Gynecology and Obstetrics (FIGO) III/IV disease. Histology was of the serous type in 41 patients (55.4%), endometrioid in 19 (25.6%), and mucinous in 14 (19%). EpCAM was overexpressed in 87.7%. Serous tumors overexpressed EpCAM significantly more often than mucinous tumors (87.8% vs. 78.6%, p=0.045); while no significant difference was noted between the other histological subgroups. EpCAM overexpression was significantly associated with a better progression free survival and higher response rates to platinum based chemotherapy (p=0.040 and p=0.048, respectively). EpCAM was identified as an independent prognostic marker for overall survival (p=0.022). CONCLUSION: Our data indicate a significant association of EpCAM overexpression with a more favorable survival in EOC-patients. Serous cancers showed a significant EpCAM overexpression compared to mucinous types. Larger multicenter analyses are warranted to confirm these findings.

Periostin as a modulator of chronic cardiac remodeling after myocardial infarction

OBJECTIVE: After acute myocardial infarction, during the cardiac repair phase, periostin is released into the infarct and activates signaling pathways that are essential for the reparative process. However, the role of periostin in chronic cardiac remodeling after myocardial infarction remains to be elucidated. Therefore, the objective of this study was to investigate the relationship between tissue periostin and cardiac variables in the chronic cardiac remodeling induced by myocardial infarction. METHODS: Male Wistar rats were assigned to 2 groups: a simulated surgery group (SHAM; n = 8) and a myocardial infarction group (myocardial infarction; n = 13). After 3 months, morphological, functional and biochemical analyses were performed. The data are expressed as means±SD or medians (including the lower and upper quartiles). RESULTS: Myocardial infarctions induced increased left ventricular diastolic and systolic areas associated with a decreased fractional area change and a posterior wall shortening velocity. With regard to the extracellular matrix variables, the myocardial infarction group presented with higher values of periostin and types I and III collagen and higher interstitial collagen volume fractions and myocardial hydroxyproline concentrations. In addition, periostin was positively correlated with type III collagen levels (r = 0.673, p = 0.029) and diastolic (r = 0.678, p = 0.036) and systolic (r = 0.795, p = 0.006) left ventricular areas. Considering the relationship between periostin and the cardiac function variables, periostin was inversely correlated with both the fractional area change (r = -0.783, p = 0.008) and the posterior wall shortening velocity (r = -0.767, p = 0.012). CONCLUSIONS: Periostin might be a modulator of deleterious cardiac remodeling in the chronic phase after myocardial infarction in rats. .

The role of micro RNAs let7c, 100 and 218 expression and their target RAS, C-MYC, BUB1, RB, SMARCA5, LAMB3 and Ki-67 in prostate cancer

OBJECTIVE: The aim of this study is to verify the expression of proteins that are controlled by miR-let7c, 100 and 218 using immunohistochemistry in tissue microarray representative of localized and metastasized the lymph nodes and bone prostate cancer. METHODS: To verify the expression of proteins that are controlled by miR-let7c (C-MYC, BUB1, RAS) 100 (SMARCA5, RB) and 218 (LAMB3) and cell proliferation (Ki-67) we used immunohistochemistry and computerized image system ImageJ MacBiophotonics in three tissue microarrays representative of localized prostate cancer and lymph node and bone metastases. miRNA expression was evaluated by qRT-PCR using 60 paraffin blocks to construct the tissue microarray representative of localized disease. RESULTS: RAS expression was increased in localized prostate cancer and bone metastases compared to the lymph nodes (p=0.017). RB showed an increase in expression from localized prostate cancer to lymph node and bone metastasis (p=0.036). LAMB3 was highly expressed in localized and lymph node metastases (p<0.001). Cell proliferation evaluated by Ki-67 showed an increase from localized prostate cancer to metastases (p<0.001). We did not found any relationship between C-MYC (p=0.253), BUB1 (p=0.649) and SMARCA5 (p=0.315) protein expression with prognosis or tumor behavior. CONCLUSION: We found that the expression of RAS, RB, LAMB3 and Ki-67 changed in the different stages of prostate cancer. Furthermore, we confirmed the overexpression of the miRNAs let7c, 100 and 218 in localized prostate cancer but failed to show the control of protein expression by the putative controller miRNAs using immunohistochemistry. .

Localización de las proteínas específicas del cemento radicular CEMP1 y CAP en células neoplásicas / Immunolocalization of cementum specific proteins CEMP1 and CAP in tumor cells

Introducción: Las proteínas CEMP1 y CAP presentes en los cementoblastos y sus progenitores contribuyen a los procesos de mineralización en tejidos del ligamento periodontal, incluyendo la migración y la proliferación de fibroblastos gingivales; sin embargo su papel y relación con procesos neoplásicos no se han estudiado a profundidad. Para lograr un mejor entendimiento de la posible contribución de estas proteínas en los procesos tumorales, particularmente en las metástasis óseas, se investigó su expresión y localización en tejidos y líneas celulares de cáncer humano. Materiales y métodos: Trece casos de cáncer de próstata y mama que desarrollaron enfermedad metastásica ósea fueron analizados por medio de inmunohistoquímica; mientras que la expresión de las proteínas en dos líneas celulares de carcinoma de próstata (PC-3) y mama (MCF-7) se estudió por medio de ensayos de Western Blot. Resultados: Los tejidos de cáncer revelaron expresión citoplasmática y ocasionalmente nuclear de CAP en células tumorales y estructuras glandulares pequeñas, así como en el citoplasma de los fibroblastos estromales adyacentes al frente de invasión tumoral. En lo correspondiente a CEMP1, su expresión se localizó en el citoplasma de las células tumorales de 5 casos, pero no en el estroma. Ensayos de Wester Blot mostraron expresión de CEMP1 en las células PC-3 y MCF-7; y de CAP en las MCF-7. Conclusiones: Los resultados muestran que las proteínas de cemento radicular CEMP1 y CAP se expresan en tejidos neoplásicos y células neoplásicas, y que posiblemente contribuyen en ciertas condiciones patológicas como el cáncer metastásico en humanos.

Introduction: CEMP1 and CAP are recognized as cementum proteins, they appear to be limited to cementoblasts and their progenitors, and participate in the mineralization process of periodontal ligament tissues, including the proliferation and migration of periodontal ligament fibroblasts. However, their contribution in neoplastic processes had not been explored. In the present study, we investigated their protein expression and localization in cancer tissues and cells. Materials and Methods: CEMP1 and CAP expressions were analyzed immunohistochemically in 13 cancer cases with bone metastasis. In addition, Wester Blot essays were use to detect expression of the proteins in the prostate (PC-3) and mama (MCF-7) cancer cell lines. Results: CAP expression was detected in all tissues examined. Strong cytoplasmatic and rarely nuclear staining was found in small tumor nests, glandular structures and, in the stromal fibroblasts at the immediate vicinity of the tumor nests. CEMP1 was found in the cytoplasm of tumor cells in 5 cases, but its expression was negative in the stromal tissues. Also, cancer lines PC-3 and MCF-7 showed CEMP1 expression; however, CAP expression was observed only in MCF-7 cells. Conclusions: The results suggest that CEMP1 and CAP are present in tissues other that cementum and possibly contribute to pathological conditions such as metastatic cancer.

Retroperitoneal Extraskeletal Ewing's Sarcoma / 대한소화기학회지

No abstract available.

Exophytic combined hepatocellular carcinoma and cholangiocarcinoma

No abstract available.

In vitro effect of glucocorticoids on nasal polyps / Efeito in vitro de glicocorticosteróides sobre pólipos nasais

Glucocorticoids are considered the main treatment option for nasal polyps, but their effect is only recently being understood. AIM: To evaluate whether fluticasone propionate (FP) inhibits the inflammatory process induced by TNF-alpha in vitro, and to assess if NF-kappaB is associated to this inhibition. STUDY DESIGN: Experimental in vitro study. MATERIALS AND METHODS: Nasal polyp fibroblasts were cultured during 24 hours. Three different concentrations of FP (1, 10 and 100 nM, added to TNF-alpha) were compared to negative (without additive) and positive (TNF-alpha) controls. Gene expression (RTQ-PCR) and protein concentration (ELISA) of VCAM-1, ICAM-1, eotaxin and RANTES were measured, as well as the nuclear translocation of NF-kappaB. RESULTS: TNF-alpha significantly increased protein concentration and RNA expression of all the studied molecules, as well as the nuclear translocation of NF-kappaB, when compared to the negative control. FP decreased these parameters in a dose-dependent manner, statistically different from positive control up to 100nM. CONCLUSIONS: FP extensively inhibited inflammatory recruiters, at both protein and RNA levels, confirming the ability of glucocorticoids to modulate the inflammatory process in nasal polyps. This inhibition was associated to decreased NF-kappaB nuclear translocation, demonstrating that this is an important mechanism of glucocorticoids action for nasal polyps.

Glicocorticoides são considerados a principal opção terapêutica para polipose nasossinusal, mas seus efeitos estão sendo descobertos apenas recentemente. OBJETIVO: Avaliar se proprionato de fluticasona (FP) inibe in vitro o processo inflamatório induzido por TNF-alfa, e se NF-kappaB está associado a esta inibição. FORMA DE ESTUDO: Experimental in vitro. MATERIAIS E MÉTODOS: Fibroblastos de pólipos nasais foram cultivados por 24 horas. Três concentrações diferentes de FP (1, 10 e 100nM, além do TNF-alfa) foram comparados a controles negativo (sem aditivo) e positivo (TNF-alfa). Expressão gênica (RTQ-PCR) concentração proteica (ELISA) de VCAM-1, ICAM-1, eotaxin e RANTES foram medidos, assim como a translocação nuclear de NF-kappaB. RESULTADOS: TNF-alfa aumentou significativamente a concentração proteica e expressão gênica de todas molé¬culas estudadas, assim como a translocação nuclear de NF-kappaB, quando comparado ao controle negativo. O FP diminuiu estes parâmetros numa forma dose-dependente, diferente estatisticamente do controle positivo até 100nM. CONCLUSÕES: O FP extensivamente inibiu os recrutadores inflamatórios, em níveis proteicos e gênicos, confirmando a habilidade dos glicocorticoides em modular o processo inflamatório na polipose nasossinusal. Esta inibição esteve associada à diminuição da translocação nuclear de NF-kappaB, demonstrando que este é um importante mecanismo de ação dos glicocorticoide na polipose nasossinusal.

GM1 improves neurofascin155 association with lipid rafts and prevents rat brain myelin injury after hypoxia-ischemia

White matter injury characterized by damage to myelin is an important process in hypoxic-ischemic brain damage (HIBD). Because the oligodendrocyte-specific isoform of neurofascin, neurofascin 155 (NF155), and its association with lipid rafts are essential for the establishment and stabilization of the paranodal junction, which is required for tight interaction between myelin and axons, we analyzed the effect of monosialotetrahexosyl ganglioside (GM1) on NF155 expression and its association with lipid rafts after HIBD in Sprague-Dawley rats, weighing 12-15 g, on day 7 post-partum (P7; N = 20 per group). HIBD was induced on P7 and the rats were divided into two groups: one group received an intraperitoneal injection of 50 mg/kg GM1 three times and the other group an injection of saline. There was also a group of 20 sham-operated rats. After sacrifice, the brains of the rats were removed on P30 and studied by immunochemistry, SDS-PAGE, Western blot analysis, and electron microscopy. Staining showed that the saline group had definite rarefaction and fragmentation of brain myelin sheaths, whereas the GM1 group had no obvious structural changes. The GM1 group had 1.9-2.9-fold more GM1 in lipid rafts than the saline group (fraction 3-6; all P < 0.05) and 0.5-2.4-fold higher expression of NF155 in lipid rafts (fraction 3-5; all P < 0.05). Injection of GM1 increased the content of GM1 in lipid rafts as well as NF155 expression and its lipid raft association in HIBD rat brains. GM1 may repair the structure of lipid rafts, promote the association of NF155 (or other important proteins) with lipid rafts, stabilize the structure of paranodes, and eventually prevent myelin sheath damage, suggesting a novel mechanism for its neuroprotective properties.

Morphine Attenuates Endothelial Cell Adhesion Molecules Induced by the Supernatant of LPS-Stimulated Colon Cancer Cells

A large reservoir of bacterial lipopolysaccharide (LPS) is available in the colon and this could promote colon cancer metastasis by enhancing tumor cell adhesion, intravasation, and extravasation. Furthermore, adhesion molecules like ICAM-1, VCAM-1, and E-selectin play important roles in the adhesion of tumor cells to endothelium. This study was designed to determine whether morphine can attenuate the expressions of adhesion molecules up-regulated by the supernatant of LPS-stimulated HCT 116 colon cancer cells (LPS-Sup). In this study, we divided to three groups by cell-growth medium of human umbilical vascular endothelial cells (HUVECs): the control group was incubated in growth factor-free endothelial medium, the Sup group was incubated in the supernatant of HCT 116 cells (Sup), and the LPS-Sup group was incubated in LPS-Sup. To observe effect of morphine to the adhesion molecules expressions in the LPS-Sup group, we co-treated morphine with LPS or added it to LPS-Sup. Adhesion molecule expressions on HUVECs in all three groups were measured during incubation period. Consquentially, ICAM-1, VCAM-1, and E-selectin expressions on HUVECs were significantly lower when morphine was co-treated with LPS than not co-treated. Thus, we suggest that morphine affects the expressions of adhesion molecules primarily by attenuating LPS stimuli on tumor cells.

Clinicopathologic Study on Combined Hepatocellular Carcinoma and Cholangiocarcinoma: with Emphasis on the Intermediate Cell Morphology

Combined hepatocellular carcinoma and cholangiocarcinoma (combined HCC-CC) is a rare subtype of primary liver cancer. We investigated the histopathologic features of transitional or intermediate areas in 21 combined HCC-CCs and immunophenotypes using different hepatic progenitor cell markers (CK7, CK19, c-kit, NCAM, and EpCAM). Major histologic findings of transitional or intermediate areas of 21 combined HCC-CCs included strands/trabeculae of small, uniform, oval-shaped cells with scant cytoplasm and hyperchromatic nuclei embedded within an abundant stroma, small cells with an antler-like anastomosing pattern, and solid nests of intermediate hepatocyte-like cells surrounded by small cells in periphery, in order of frequency. The intermediate area of one tumor was composed predominantly of spindle cells arranged in short fascicles. Immunophenotype of tumor cells with intermediate morphology suggested a progenitor cell origin for this tumor. Clinical findings of combined HCC-CC showed a closer resemblance with those of HCC than those of CC. In univariate analysis, tumor size, TNM stage, and serum alpha-fetoprotein levels showed a significant association with poor patient survival. Serum alpha-fetoprotein level was an independent prognostic indicator in multivariate analysis. In conclusion, an awareness of the clinicopathologic features, specifically the various morphologic features of intermediate areas in this tumor, is essential for prevention of potential misdiagnosis as another tumor.