Efecto de la epigenética en la artritis reumatoide / Effect of epigenetics on rheumatoid arthritis

La artritis reumatoide es una enfermedad autoinmune e inflamatoria que afecta de manera predominante a las articulaciones diartrodiales. En esta patología los factores ambientales o conductuales pueden actuar en sinergia con la predisposición genética, acelerando el inicio y la gravedad de la enfermedad. Este vínculo entre el medio ambiente y el genoma está mediado por marcas epigenéticas en el ácido desoxirribonucleico, incluyendo su metilación, la modificación de histonas y la regulación mediada por ácido ribonucleico no codificante. La epigenética puede generar cambios fenotípicos hereditarios, que no están determinados por modificaciones en la secuencia del ácido desoxirribonucleico y, en consecuencia, son reversibles. Por lo tanto la dieta, los medicamentos y otros factores ambientales, tendrían la capacidad de modularlos. La identificación de una desregulación epigenética específica, puede ofrecer una mayor comprensión de la fisiopatología de la enfermedad e influenciar positivamente en la prevención, diagnóstico y desarrollo de nuevas dianas terapéuticas.

Rheumatoid arthritis is an autoimmune and inflammatory disease that predominantly affects the diarthrodial joints. In this pathology, environmental or behavioral factors can act in synergy with genetic predisposition, accelerating the onset and severity of the disease. This link between the environment and the genome is mediated by epigenetic marks on deoxyribonucleic acid, including its methylation, histone modification, and noncoding ribonucleic acid-mediated regulation. Epigenetics can generate heritable phenotypic changes, which are not determined by modifications in the deoxyribonucleic acid sequence and are therefore reversible. Therefore, diet, medications and other environmental factors would have the ability to modulate them. The identification of a specific epigenetic dysregulation can offer a better understanding of the pathophysiology of the disease and positively influence the prevention, diagnosis and development of new therapeutic targets.

Expression of monoubiquitination histone H2B in esophageal carcinoma and its clinical significance / 肿瘤研究与临床

Objective:To investigate the expression of monoubiquitination histone H2B (H2Bub) in esophageal cancer tissues and its correlation with the prognosis of patients.Methods:A total of 75 patients who underwent thoracic esophagectomy in Shanxi Province Cancer Hospital from May 2010 to December 2015 were selected. The expression of H2Bub protein in esophageal carcinoma and para-carcinoma tissues was detected by using immunohistochemical method. The relationship between H2Bub expression level and the clinicopathological characteristics was analyzed, Kaplan-Meier method was used to analyze the relationship H2Bub expression level and the survival.Results:H2Bub was positively expressed in esophageal carcinoma and para-carcinoma tissues, and weakly positive expressed H2Bub was found in para-carcinoma tissues, while not found in esophageal carcinoma tissues. The strongly positive expression rate of H2Bub in esophageal carcinoma tissues was higher than that in para-carcinoma tissues [84.0% (63/75) vs. 22.7% (17/75), χ2 = 34.68, P < 0.001]. Compared with para-carcinoma tissues, 64.0% (48/75) of H2Bub expression level in carcinoma tissues was up-regulated, and 2.7% (2/75) of H2Bub expression level was down-regulated. The up-regulated expression of H2Bub in esophageal carcinoma tissues compared with para-carcinoma tissues was not related with the gender, age, tumor diameter, lymph node metastasis and T staging (all P > 0.05). The proportion of patients with up-regulated expression of H2Bub in poorly differentiated carcinoma tissues was lower than that in moderately and highly differentiated carcinoma tissues [43.8% (7/16) vs. 66.7% (34/51), 87.5% (7/8), P = 0.037]. The median overall survival time was 70 months (95% CI 45-95 months) and 68 months (95% CI 54-82 months), respectively in 12 esophageal carcinoma patients with moderately positive expressed H2Bub and 63 esophageal carcinoma patients with strongly positive expressed H2Bub, and the difference was statistically significant ( P = 0.606). Among 48 patients with up-regulated expression of H2Bub in esophageal carcinoma tissues compared with para-carcinoma tissues, the median overall survival time of poorly differentiated esophageal carcinoma group (7 cases) was shorter than that of highly differentiated (7 cases) and moderately differentiated (34 cases) esophageal carcinoma group [36 months (95% CI 24-37 months) vs. 68 months (95% CI 38-98 months), 68 months (95% CI 44-91 months)], and the differences were statistically significant (all P < 0.05). Conclusions:The expression level of H2Bub in esophageal carcinoma tissues is up-regulated compared with that in para-carcinoma tissues. The up-regulated H2Bub expression level of patients with poorly differentiated esophageal carcinoma with poor prognosis is obvious.

Epigenetics and drug resistance in lung cancer / 国际肿瘤学杂志

Lung cancer has the highest mortality rate among malignant tumors, and drug resistance in lung cancer treatment is currently a major challenge for clinical practitioners. Recent studies have found that epigenetics is closely linked to drug resistance in lung cancer, especially in DNA methylation, histone modifications and non-coding RNA regulation. By exploring the mechanisms affecting drug resistance in lung cancer through the above three features and exploring drugs that can effectively improve drug resistance in lung cancer in response to the mechanisms, it can provide ideas for solving the problems related to drug resistance in lung cancer in clinical work and provide a reference basis for prognostic assessment of lung cancer patients.

Activating effect of extracellular histones on heparinase in chlorine-induced acute lung injury / 中国职业医学

OBJECTIVE: To investigate the induction and activation of heparinase by extracellular histones in acute respiratory distress syndrome(ARDS) induced by chlorine in mice.METHODS: The specific pathogen free adult male C57 BL/6 mice were randomly divided into control group, chlorine injured group, histone injured group, anti-histone antibody group and heparinase inhibitor group, with six mice in each group.The mice in the control group and histone injured group were exposed to clean air, and the mice in the other three groups were exposed to chlorine gas at a dose of 580.0 mg/m~3 for 30 minutes by systemic dynamic inhalation.Mice in the histone injured group were injected with 50 mg/kg body weight calf thymus histone by tail vein.One hour before exposure, mice in the anti-histone antibody group were pretreated with 20 mg/kg body weight anti-histone H4 antibody by tail vein injection, and mice in the heparinase inhibitor group were injected with 2 mg/kg body weight OGT2115(heparinase inhibitor). The other three groups were given equal volume of 0.9% sodium chloride solution by tail vein injection. After 24 hours of exposure, arterial blood was collected for blood gas analysis and the lung tissue was collected for histopathological examination. The protein level of heparinase in lung tissue were detected using enzyme-linked immunosorbent assay, and the activity of heparinase were detected by measuring the product of heparan degradation. The protein expression of pro-heparinase and active heparinase were detected by Western blotting.RESULTS: The dyspnea developed of mice in the chlorine injured group and histone injured group, diffuse inflammation occurred in lung tissue, the oxygenation index in arterial blood decreased(all P<0.05), and the protein level and activity of heparinase in lung tissue, as well as the relative expression of pro-heparinase and active heparinase were increased compared with the control group(all P<0.05). The dyspnea, hypoxemia and acute lung injury of mice in the anti-histone antibody group were alleviated, and the protein level of heparinase in lung tissue, as well as the relative expression levels of pro-heparinase and active heparinase were decreased(all P<0.05), compared with chlorine injury group and histone injury group.The dyspnea, hypoxemia and acute lung injury were alleviated in the heparinase inhibitor group, and the activity of heparinase and the relative expression of pro-heparinase in the lung tissue were decreased compared with the chlorine injury group(all P<0.05). CONCLUSION: During the occurrence and development of chlorine-induced ARDS in mice, extracellular histones aggravate lung injury by inducing the expression and activation of heparinase. Acute lung injury can be alleviated by inhibiting the expression and activation of heparinase.

Trampas extracelulares de neutrófilos en sepsis / Neutrophil extracellular traps in sepsis / Armadilhas extracelulares de neutrófilos na sepse

Resumen: Introducción: Los neutrófilos son uno de los componentes celulares más importantes de la inmunidad innata, debido a que estas células se reclutan rápidamente a los sitios de infección y pueden eliminar patógenos por múltiples métodos. En diferentes entidades, de entre las que se destaca la sepsis, los neutrófilos mejoran sus propiedades antimicrobianas mediante la liberación de trampas extracelulares de neutrófilos (TEN), constituidas por cromatina, histonas y proteínas granulares, proceso que es conocido por NETosis. Objetivo: El objetivo de esta revisión es examinar los conceptos actuales, relacionados con los mecanismos subyacentes de la formación de TEN y su impacto en sepsis. Conclusión: La desregulación de la NETosis causada por sepsis puede tener efectos deletéreos en sepsis, destacando inflamación, trombosis y disfunción multiorgánica.

Abstract: Introduction: Neutrophils are one of the most important cellular components of innate immunity because these cells are rapidly recruited to sites of infection and can eliminate pathogens by multiple methods. In different entities from which sepsis disables neutrophils improve their antimicrobial properties by releasing neutrophils extracellular traps (NET), constituted by chromatin, histones and granular proteins, a process that is known by NETosis. Objective: The objective of this paper is to review current concepts related to the underlying mechanisms of the formation of NETs, as well as the beneficial and harmful effects of them. Conclusions: The dysregulation of NETosis caused by sepsis can have detrimental effects that cause inflammation, thrombosis and multi-organ failure.

Resumo: Introdução: Os neutrófilos são um dos componentes celulares mais importantes da imunidade inata, porque essas células se recrutam rapidamente para locais de infecção e podem matar patógenos por vários métodos. Em diferentes entidades das quais se destaca a sepse, os neutrófilos melhoram suas propriedades antimicrobianas liberando as armadilhas extracelulares de neutrófilos (del inglés: neutrophil extracelular traps, NET), compostas por cromatina, histonas e proteínas granulares, um processo conhecido como NETosis. Objetivo: O objetivo desta revisão é revisar os conceitos atuais relacionados aos mecanismos subjacentes à formação das NET e seu impacto na sepse. Conclusão: A desregulação da NETosis causada pela sepse pode ter efeitos deletérios na sepse, destacando inflamação, trombose e disfunção de múltiplos órgãos.

The conserved aspartate in motif III of b family AdoMet-dependent DNA methyltransferase is important for methylation

S-adenosyl-L-methionine (AdoMet)-dependent methyltransferases (MTases) are involved in diverse cellularfunctions. These enzymes show little sequence conservation but have a conserved structural fold. The DNAMTases have characteristic motifs that are involved in AdoMet binding, DNA target recognition and catalysis.Motif III of these MTases have a highly conserved acidic residue, often an aspartate, whose functionalsignificance is not clear. Here, we report a mutational study of the residue in the b family MTase of the Type IIIrestriction-modification enzyme EcoP15I. Replacement of this residue by alanine affects its methylationactivity. We propose that this residue contributes to the affinity of the enzyme for AdoMet. Analysis of thestructures of DNA, RNA and protein MTases reveal that the acidic residue is conserved in all of them, andinteracts with N6 of the adenine moiety of AdoMet. Interestingly, in the SET-domain protein lysine MTases,which have a fold different from other AdoMet-dependent MTases, N6 of the adenine moiety is hydrogenbonded to the main chain carbonyl group of the histidine residue of the highly conserved motif III. Our studyreveals the evolutionary conservation of a carbonyl group in DNA, RNA and protein AdoMet-dependentMTases for specific interaction by hydrogen bond with AdoMet, despite the lack of overall sequenceconservation

Glioma difuso de línea media H3K27M positivo en adulto. Caso clínico / H3K27 positive diffuse midline glioma. Report of one case

The 2016 WHO Classification of Tumours of the Central Nervous System incorporates a new diagnostic entity: the mutant diffuse midline glioma H3K27, a tumor with a characteristic location and special molecular biology. We report the case of a 51-year-old male patient with progressive diplopia. The imaging study showed a mesencephalic tumor; the stereotacic biopsy disclosed an Anaplastic Astrocytoma Isocitrate dehydrogenase (IDH) wild type. The molecular study concludes H3K27 mutation. The patient was treated with radiotherapy with concurrent and adjuvant chemotherapy (temozolomide) with partial recovery of the diplopia.

Angiotensin II and TGF-β1 Induce Alterations in Human Amniotic Fluid-Derived Mesenchymal Stem Cells Leading to Cardiomyogenic Differentiation Initiation

BACKGROUND AND OBJECTIVES: Human amniotic fluid-derived mesenchymal stem cells (AF-MSCs) may be a valuable source for cardiovascular tissue engineering and cell therapy. The aim of this study is to verify angiotensin II and transforming growth factor-beta 1 (TGF-β1) as potential cardiomyogenic differentiation inducers of AF-MSCs. METHODS AND RESULTS: AF-MSCs were obtained from amniocentesis samples from second-trimester pregnant women, isolated and characterized by the expression of cell surface markers (CD44, CD90, CD105 positive; CD34 negative) and pluripotency genes (OCT4, SOX2, NANOG, REX1). Cardiomyogenic differentiation was induced using different concentrations of angiotensin II and TGF-β1. Successful initiation of differentiation was confirmed by alterations in cell morphology, upregulation of cardiac genes-markers NKX2-5, TBX5, GATA4, MYH6, TNNT2, DES and main cardiac ion channels genes (sodium, calcium, potassium) as determined by RT-qPCR. Western blot and immunofluorescence analysis revealed the increased expression of Connexin43, the main component of gap junctions, and Nkx2.5, the early cardiac transcription factor. Induced AF-MSCs switched their phenotype towards more energetic and started utilizing oxidative phosphorylation more than glycolysis for energy production as assessed using Agilent Seahorse XF analyzer. The immune analysis of chromatin-modifying enzymes DNMT1, HDAC1/2 and Polycomb repressive complex 1 and 2 (PRC1/2) proteins BMI1, EZH2 and SUZ12 as well as of modified histones H3 and H4 indicated global chromatin remodeling during the induced differentiation. CONCLUSIONS: Angiotensin II and TGF-β1 are efficient cardiomyogenic inducers of human AF-MSCs; they initiate alterations at the gene and protein expression, metabolic and epigenetic levels in stem cells leading towards cardiomyocyte-like phenotype formation.

Histone variants: critical determinants in tumour heterogeneity / 医学前沿

Malignant cell transformation could be considered as a series of cell reprogramming events driven by oncogenic transcription factors and upstream signalling pathways. Chromatin plasticity and dynamics are critical determinants in the control of cell reprograming. An increase in chromatin dynamics could therefore constitute an essential step in driving oncogenesis and in generating tumour cell heterogeneity, which is indispensable for the selection of aggressive properties, including the ability of cells to disseminate and acquire resistance to treatments. Histone supply and dosage, as well as histone variants, are the best-known regulators of chromatin dynamics. By facilitating cell reprogramming, histone under-dosage and histone variants should also be crucial in cell transformation and tumour metastasis. Here we summarize and discuss our knowledge of the role of histone supply and histone variants in chromatin dynamics and their ability to enhance oncogenic cell reprogramming and tumour heterogeneity.

Activation of lung endothelial cells by extracellular histone in mice with acute respiratory distress syndrome / 中华劳动卫生职业病杂志

Objective@#To observe the changes of extracellular histones and pulmonary microvascular endothelial cells, and study the activating role of extracellular histones to pulmonary microvascular endothelial cells in the pathogenesis of acute respiratory distress syndrome (ARDS) .@*Methods@#The correlation of the severity of acute lung injury with extracellular histones and pulmonary endothelial damage was studied through mice model, and acute lung injury was produced by aspiration of different concentrations of hydrochloric acid (0.01、0.1、0.3 and 0.5 mol/L, 2 ml/kg). Tumor necrosis factor-α (TNF-α), soluble thrombomodulin (sTM) and lung pathological change were measured. The pro-inflammatory role of extracellular histones was tested by injecting calf thymus histones (CTH) or specific anti-H4 antibody through tail vein. The direct activating role of extracellular histones to pulmonary microvascular endothelial cells was studied through pulmonary endothelial model.@*Results@#The extracellular histones in plasma were increased obviously 6h after aspiration of different concentrations of hydrochloric acid in mice. A positive correlation was seen between extracellular histones and concentrations of aspirated hydrochloric acid (r=0.9180, P<0.05). The sTM in plasma also showed a positive correlation with concentrations of aspirated hydrochloric acid (r=0.8701, P<0.05). Merely administering CTH could not only increase TNF-α and sTM in plasma but also cause obvious lung injury, while specific anti-H4 antibody could relieve the inflammation and lung damage caused by CTH. Extracellular histones could directly damage pulmonary endothelial cells to release sTM in pulmonary endothelial model in vitro, while anti-H4 antibody could protect the endothelial cells.@*Conclusion@#Extracellular histones are the key endogenic inflammatory mediators during the pathogenesis of ARDS caused by aspiration of hydrochloric acid, which could promote inflammation by directly activating pulmonary endothelial cells.

Prediction value of plasma histone in prognosis of sepsis patients / 中华危重病急救医学

Objective To explore the value of plasma histones in predicting the prognosis of sepsis patients. Methods The patients with sepsis admitted to intensive care unit (ICU) of Subei People's Hospital of Jiangsu Province Affiliated to Yangzhou University from May 2016 to June 2018 were enrolled as the research subjects, and healthy volunteers were selected as healthy control at the same period. The plasma levels of histones, cardiac troponin I (cTnI), N-terminal pro-brain natriuretic peptide (NT-proBNP), sequential organ failure assessment (SOFA) score, lactate (Lac), procalcitonin (PCT) on admission 24 hours, and use of vasoconstrictor agents, the length of ICU stay and ICU mortality were recorded. The patients were divided into survival group and death group according to the prognosis, and the differences of each index between the two groups were compared. Multivariate binary Logistic regression analysis was carried out to identify the independent risk factors of death. The correlation between histone and the levels of cTnI, NT-proBNP, PCT and Lac was analyzed. The value of plasma histone, cTnI, NT-proBNP, PCT and Lac in predicting the prognosis of patients was analyzed by receiver operating characteristic (ROC) curve. According to the threshold value of histone in predicting prognosis, the patients were divided into two groups, and the differences of various indicators between the two groups were compared. Results ① A total of 93 sepsis patients were included, with 29 cases of ICU death, and the mortality was 31.2%. ② Compared with the healthy control group, histones, cTnI, NT-proBNP were significant increased, besides, histones, cTnI in the death group were further increased compared with the survival group;in addition, SOFA, proportion of vasoconstrictor use were also significant higher than those in the survival group [histones (mg/L): 0.33 (0.28,0.45) vs. 0.22 (0.17,0.29), cTnI (μg/L): 0.25±0.13 vs. 0.20±0.08, SOFA: 11 (8, 12) vs. 9 (8, 11), the rate of vasopressor use: 93.1% (27/29) vs. 68.8% (44/64), all P < 0.05]. Statistically significant indicators between the two groups were included in multivariate binary Logistic regression analysis. The result showed that the independent risk factors affecting the prognosis of patients were the rate of vasopressor use [odds ratio (OR) = 5.277, P = 0.043] and the level of histone (OR = 79.244, P = 0.036). ③ The plasma histone level were positively correlated with cTnI (r = 0.577, P = 0.000), SOFA (r = 0.469, P = 0.000), NT-proBNP (r = 0.349, P = 0.001) and Lac (r = 0.357, P = 0.000), while there was no significant correlation between histone and PCT (r = 0.133, P = 0.205). ④ ROC curve analysis showed that the area under ROC curve (AUC) of histone predicting prognosis was 0.769 (P = 0.000); when the cut-off point was 0.30 mg/L, the sensitivity and specificity were 72.4% and 81.2% respectively. The AUC of SOFA score was 0.653 (P = 0.018), and the sensitivity and specificity were 58.6% and 70.3% respectively when the cut-off point was 10.50; while cTnI, NT-proBNP, Lac and PCT had little value in predicting the prognosis of patients. ⑤ Compared with the group with histone level lower than 0.3 mg/L, the group with histones level greater than 0.3 mg/L had higher SOFA score, more doses of vasopressor, higher cTnI, NT-proBNP, Lac and PCT levels, and higher ICU mortality [SOFA: 11 (10, 12) vs. 9 (8, 10), use of vasopressor: 84.8% (28/33) vs. 76.7% (46/60), cTnI (μg/L): 0.28 (0.19, 0.32) vs. 0.18 (0.12, 0.22), NT-proBNP (ng/L): 3 624.0 (2 800.0, 5 260.0) vs. 2 512.0 (1 361.8, 3 590.8), Lac (mmol/L): 2.25 (1.85, 3.50) vs. 1.60 (1.25, 2.35), PCT (μg/L): 2.10 (1.30, 4.03) vs. 1.60 (1.26, 2.33), ICU mortality: 48.5% (16/33) vs. 21.7% (13/60), all P < 0.05], while no statistical difference in the length of ICU stay was found. Conclusions The independent risk factors for ICU mortality of sepsis patients were high histone level and the use of vasopressor. Plasma histone can be regarded as an indicator in predicting the prognosis of patients with sepsis.

Research progress about neutrophil extracellular trapping network and histone‐mediated thrombogene‐sis／ / 心血管康复医学杂志

:Platelets and neutrophils are now considered the key factor to thrombus initiation and progression .The present article describes the latest discovery of neutrophil extracellular trapping network (NETs) in thrombogenesis and unknown field to be explore ,including biological process of NET formation (NETosis) ,and how extracellular release of DNA and histone of NETs help coagulation and platelet aggregation .Cell biology of NETosis is still ac‐tively characterized ,which may provide new specific inhibition therapy insights .

The comparison of heparan sulfate and its fragments on the protection against extracellular histones during the pathogenesis of acute respiratory distress syndrome / 中华劳动卫生职业病杂志

Objective@#In order to explore the role of heparan sulfate (HS) during the pathogenesis of acute respiratory distress syndrome (ARDS) , the protective effect of HS and its fragments against extracellular histones was compared.@*Methods@#Calf thymus histones (CTH) were injected via femoral vein to induce ARDS in rats. HS, HS fragments or saline was intraperitoneally injected (10mg/kg, Q6h, 24h) to test the protective effect against CTH. The ratio of wet/dry lung weight, protein content in bronchoalveolar lavage fluid (BALF) , total leukocyte and neutrophil count in BALF were measured.@*Results@#After CTH injection, the ratio of wet/dry lung weight (5.7±0.95) was much higher than the saline control group (3.1±0.15). The protein content (0.47±0.086mg/ml) , total leukocyte[ (97.4±15.6l) ×104/ml] and neutrophil (18±3.4/LPF) in BALF were obviously increased compared with the saline control group. The intervention of HS evidently decreased ratio of wet/dry lung weight (4.2±0.41) , protein content[ (0.26±0.019) mg/ml], leukocyte[ (61.3±5.74) ×104/ml] and neutrophil (12±1.8/LPF) in BALF. HS fragments also decreased ratio of wet/dry lung weight, protein content, leukocyte and neutrophil count in BALF though the strength was much less than HS.@*Conclusion@#HS and its fragments could provide protection against extracellular histones during the pathogenesis of ARDS. For the protective effect full length HS was much better than HS fragments.

Expression and clinical significance of PHH3 and β-catenin in the tissue of non-small cell lung cancer / 中国基层医药

Objective To investigate the expression and clinical significance of PHH 3 and β-catenin in non-small cell lung cancer ( NSCLC) .Methods 78 cases of NSCLC tissue removed from June 2013 to June 2017 in Rizhao Central Hospital were collected .The expression of PHH3 and β-catenin protein in NSCLC and 30 cases of normal lung tissues were detected by immunohistochemical MaxVision method .The relationship between the expres-sion of PHH3 andβ-catenin protein was analyzed .The relationship between the expression of PHH 3,β-catenin and the clinicopathological features of NSCLC was analyzed .Results The positive rate of PHH3 in NSCLC tissues ( 57/78,73.1％) was significantly higher than that in normal lung tissues (2/30,6.7％)(χ2 =38.553,P<0.05).The expression level of PHH3 was significantly correlated with tumor differentiation (χ2 =5.248,P<0.05),lymph node metastasis(χ2 =7.747,P<0.05),but it was not related to the age (χ2 =0.209,P>0.05),gender(χ2 =0.033,P>0.05) and pathological type (χ2 =0.190,P>0.05).The positive rate of β-catenin in NSCLC tissues (54/78, 69.2％) was significantly higher than that in normal lung tissues (3/30,10.0％) (χ2 =30.499,P <0.05).The expression level of β-catenin was significantly correlated with tumor differentiation (χ2 =4.934,P<0.05),lymph node metastasis(χ2 =8.098,P<0.05),but it was not related to the age (χ2 =0.006,P>0.05),gender(χ2 =0.595,P>0.05) and pathological type (χ2 =0.071,P>0.05).The expression of PHH3 was positively correlated withβ-catenin(r=0.597,P<0.05).Kaplan-Meier analysis showed that PHH3 and β-catenin expression was negatively correlated with prognosis .Conclusion The overexpression of PHH3 and β-catenin in NSCLC may be related to the development ,invasion and metastasis of NSCLC ,which is expected to be an indicator of NSCLC ,and to predict invasion and metastasis .

Effect of resuscitation with hypertonic sodium chloride hydroxyethyl starch 40 injection mixed with suberoylanilide hydroxamic acid on oxidative stress responses and histone acetylation in lung tissues in a rat model of lethal hemorrhagic shock after entering high altitude for the first time / 中华麻醉学杂志

Objective To evaluate the effect of resuscitation with hypertonic sodium chloride hydroxyethyl starch 40 injection (HSH40) mixed with suberoylanilide hydroxamic acid (SAHA) on oxidative stress responses of lung tissues and histone acetylation in a rat model of lethal hemorrhagic shock after entering high altitude for the first time.Methods Forty-five healthy male Wistar rats,aged 3-4 months,weighing 250-300 g,were transported from the breeding area at altitude 1500 m to the experimental area at altitude 3 780 m.The rats were divided into 3 groups (n=15 each) using a random number table:sham operation group (group Sham),hemorrhagic shock group (group HS),and resuscitation with HSH40 mixed with SAHA group (group HSH/SAHA).Lethal hemorrhagic shock was induced by removing 40％ of blood volume from the left femoral artery at a constant speed within 10 min,followed by removing 15％ of blood volume from the right femoral vein at a constant speed within 50 min.Only cannulation was performed,and the rats received no blood letting or resuscitation in group Sham.The animals were resuscitated via the right femoral artery after successful establishment of the model,SAHA 7.5/Kg dissolved in HSH40 4 ml/kg was infused within 5 min in group HSH+SAHA.Immediately before blood letting,immediately after blood letting and at 3 h after resuscitation (at the time of death for the rats survived less than 3 h),arterial blood samples were obtained for blood gas analysis,and pH value,partial pressure of arterial carbon dioxide (PaCO2),partial pressure of arterial oxygen (PaO2) and arterial oxygen saturation (SaO2) were recorded.The rats were sacrificed after blood samples were collected from the abdominal aorta at 3 h after resuscitation (at the time of death for the rats died within 3 h after resuscitation),and lungs were removed for examination of the pathologic changes which were scored (with a light microscope) and for determination of wet to dry weight ratio (W/D ratio),activity of superoxide dismutase (SOD) and content of malondialdehyde (MDA) and expression of histone H3 acetylation at lysine 9 (Ac-H3K9) in lung tissues (by Western blot).Results Compared with group Sham,the lung injury score,W/D ratio and content of MDA were significantly increased,and the activity of SOD was decreased in HS and HSH+SAHA groups,pH value and PaCO2 were significantly decreased and PaO2 and SaO2 were increased immediately after blood letting and at 3 h after resuscitation in group HS,and PaO2 and SaO2 were significantly increased immediately after blood letting and at 3 h after resuscitation,pH value and PaCO2 were decreased immediately after blood letting,and the expression of Ac-H3K9 was up-regulated in group HSH+SAHA (P＜0.05).Compared with group HS,pH value,PaCO2,PaO2 and SaO2 were significantly increased at 3 h after resuscitation,the lung injury score,W/D ratio and content of MDA were decreased,the activity of SOD was increased,and the expression of Ac-H3K9 was up-regulated in group HSH+SAHA (P＜0.05).Conclusion The mechanism by which resuscitation with HSH40 mixed with SAHA exerts lung protection may be related to inhibiting oxidative stress responses and histone acetylation in lung tissues in a rat model of lethal hemorrhagic shock after entering high altitude for the first time.

Role of spinal histone acetylation in persistent postoperative pain in rats / 中华麻醉学杂志

Objective To evaluate the role of spinal histone acetylation in persistent postoperative pain in rats. Methods Pathogen?free healthy male Sprague?Dawley rats, weighing 200-250 g, aged 2 months, in which intrathecal catheters were implanted at the lumbar level according to an improved method, were used in the study. Eighty?four rats, in which intrathecal catheters were successfully implanted, were divided intoⅠ-Ⅵgroups(n=14 each)using a random number table. Artificial cerebrospinal fluid 20 μl was intrathecally administered at 1, 2, 3 and 4 days before operation and 1 day after operation inⅠandⅣgroups. At 1, 2, 3 and 4 days before operation and 1 day after operation, dimethyl sulfoxide 10 μl and SAHA(50 μg∕10μl)were intrathecally injected inⅡandⅤgroups and inⅢandⅥgroups, respective?ly, followed by artificial cerebrospinal fluid(10 μl)flush after each injection. Rats underwent sham oper?ation inⅠ?Ⅲ groups. Persistent postoperative pain was evoked by skin∕muscle incision and traction in Ⅳ?Ⅵ groups. The mechanical paw withdrawal threshold(MWT)was measured at 1 day before operation(T0)and 1, 3, 7, 14 and 21 days after operation(T1?5). Four rats were sacrificed in each group after measurement of MWT at T4, and the lumbar segments(L4?6)of the spinal cord were removed for determi?nation of the expression of acetylated histone H3(Ac?H3)and Ac?H4 by Western blot. Results There was no significant difference in each index amongⅠ?Ⅲ groups(P>0.05). Compared with group Ⅰ, the MWT was significantly decreased at T2?5, and the expression of Ac?H3 and Ac?H4 was down?regulated at T4 in group Ⅳ(P<0.05). Compared with group Ⅳ, the MWT was significantly increased at T2?5, and the expression of Ac?H3 and Ac?H4 was up?regulated at T4in group Ⅵ(P<0.05). Conclusion Histone acetylation is involved in the development and maintenance of persistent postoperative pain in rats.

Induced pluripotent stem cells reprogramming: Epigenetics and applications in the regenerative medicine / Células-tronco de pluripotência induzida: papel da epigenética na reprogramação e sua aplicabilidade clínica

Summary Induced pluripotent stem cells (iPSCs) are somatic cells reprogrammed into an embryonic-like pluripotent state by the expression of specific transcription factors. iPSC technology is expected to revolutionize regenerative medicine in the near future. Despite the fact that these cells have the capacity to self-renew, they present low efficiency of reprogramming. Recent studies have demonstrated that the previous somatic epigenetic signature is a limiting factor in iPSC performance. Indeed, the process of effective reprogramming involves a complete remodeling of the existing somatic epigenetic memory, followed by the establishment of a "new epigenetic signature" that complies with the new type of cell to be differentiated. Therefore, further investigations of epigenetic modifications associated with iPSC reprogramming are required in an attempt to improve their self-renew capacity and potency, as well as their application in regenerative medicine, with a new strategy to reduce the damage in degenerative diseases. Our review aimed to summarize the most recent findings on epigenetics and iPSC, focusing on DNA methylation, histone modifications and microRNAs, highlighting their potential in translating cell therapy into clinics.

Resumo As células-tronco de pluripotência induzida (CTPI) ou do inglês induced pluripotent stem cells (iPSCs) são células somáticas reprogramadas para o estado embrionário por meio da expressão de fatores ectópicos de transcrição específicos, tornando-as um alvo promissor para a medicina regenerativa. Apesar das CTPI compartilharem características embrionárias, como pluripotência e capacidade de autorrenovação, elas possuem uma baixa eficiência de reprogramação, sendo a memória epigenética uma das principais barreiras nesse processo. A epigenética é caracterizada por alterações reversíveis e herdáveis no genoma funcional que não alteram a sequência de nucleotídeos do DNA. Dentre as diferentes modificações epigenéticas, destacam-se metilação de DNA, alterações em histonas e microRNA. Atualmente, sabe-se que o processo de reprogramação efetivo das CTPI envolve um completo remodelamento da memória epigenética somática existente, seguido pelo estabelecimento de uma "assinatura epigenética" que esteja de acordo com o novo tipo de célula a ser diferenciada. Modificações epigenéticas personalizadas são capazes de melhorar o rendimento e a efetividade das CTPI geradas, abrindo uma nova perspectiva para a terapia celular. Nesta revisão reunimos as principais informações sobre os fatores epigenéticos que afetam a reprogramação das CTPI, bem como seus benefícios na aplicação da terapia celular.

Histone modification in breast cancer / 国际肿瘤学杂志

Epigenetic regulation plays an important role in breast cancer research.The epigenetic modification of histone mainly affects the occurrence and development of breast cancer by adding and removing methyl,acetyl and phosphate groups under the action of various enzymes.Because of its reversible regulatory process,it can provide advantage for the affected region to return to the normal genome state.Clinically,this can be used to develop a variety of drugs for patient diagnosis and treatment and to provide treatment targets.

Effect of heparin pretreatment on the level of neutrophil extracellular traps of serum and lung tissue in septic mice / 中华危重病急救医学

Objective To investigate the influence of heparin pretreatment on serum and lung tissue level of neutrophil extracellular traps (NETs) in septic mice model and its molecular mechanism.Methods Ninety male C57BL/6J mice were randomly divided into control group (n = 30), lipopolysaccharides (LPS) group (n = 30, 30 mg/kg LPS in 100μL normal saline was intraperitoneally injected) and LPS+heparin group (n = 30, 8 U of heparin in 20μL normal saline was subcutaneously injected 30 minutes before the injection of LPS). Six hours later of LPS injection, blood was collected and lung tissue was harvested. Enzyme linked immunosorbent assay (ELISA) was used to assess the concentration of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and histones 2AX (H2AX), neutrophil elastase (NE), which reflected NETs concentration. PicoGreen fluorescent dyes was used to detect serum circulating free DNA (cf-DNA/NETs) concentration. The protein expression levels of H2AX and NE in lung tissue were examined by Western Blot.Results The serum concentrations of TNF-α, IL-6, H2AX, NE, cf-DNA/NETs, and the protein expression levels of H2AX and NE in lung tissue of septic mice were significantly higher than those of control group [TNF-α (ng/L): 133.0±14.1 vs. 2.7±1.0, IL-6 (ng/L): 3911.2±189.2 vs. 298.9±52.5, H2AX (ng/L): 545.5±40.0 vs. 21.9±8.3, NE (μg/L): 6.48±0.12 vs. 0.47±0.15, cf-DNA/NETs (μg/L): 846.3±137.5 vs. 152.7±36.4, H2AX protein (gray value): 1.14±0.09 vs. 0.68±0.04, NE protein (gray value): 0.56±0.03 vs. 0.32±0.04, allP 0.05).Conclusion Heparin pretreatment could significantly decrease the level of NETs in serum and lung tissue, and can be the potential mechanism of its organ protection in sepsis.

Relationship between O6-methylguanine-DNA methyltransferase gene regulated by H3K36me3 and DNA damage induced by arsenic in HaCaT Cells / 中华地方病学杂志

Objective To observe the influences of NaAsO2 on H3K36me3 modifications,mRNA transcription of O6-methylguanine-DNA methyltransferase gene (MGMT) in HaCaT cells,and to explore the relationship between the transcription of MGMT gene regulated by H3K36me3 and DNA damage induced by arsenic,in order to provide new ideas and scientific basis for prevention and intervention of arsenism.Methods HaCaT cells were treated with 1.25,2.50,5.00 and 10.00 μmol/L NaAsO2 for 24 h,and were also treated with 10.00 μmol/L NaAsO2 for 6,12 and 24 h.HaCaT cells that treated with 0.00 pmol/L NaAsO2 and 0 h were used as blank control group.The degree of DNA damage in peripheral blood cells was detected by single cell gel electrophoresis.The level of H3K36me3 modifications was detected using Western blotting.Quantitative real-time polymerase chain reaction was used to detect the mRNA levels of MGMT gene.Quantitative chromatin immuno-precipitation was used to detect the level of H3K36me3 modifications in the coding regions (ChIP1 and ChIP2) of MGMT gene.Results ①Among the groups of HaCaT cells treated with 2.50,5.00 and 10.00 μmol/L NaAsO2,the levels of tail DNA％ (11.83 ± 1.15,16.85 ± 2.52,24.23 ± 2.75) and olive tail moment (OTM,10.90 ± 1.13,16.19 ± 2.26,23.83 ± 2.79)were significantly increased compared with those of the control group (0.00 μmol/L,2.40 ± 0.51,2.26 ± 0.40,all P ＜ 0.05).After treated with 10.00 μmoFL NaAsO2 for 12 and 24 h,compared with the control group (0 h,3.66 ± 1.02,3.38 ± 1.00),the degrees of tail DNA％ (15.51 ± 1.92,24.18 ± 2.42) and OTM (13.58 ± 2.04,23.14 ± 2.11)were significantly increased (all P ＜ 0.05).②Compared with the control group (0.00 μmol/L,100.00 ± 0.00),the levels of H3K36me3 modifications (60.59 ± 9.75,57.82 ± 11.28,39.45 ± 7.09) were lower at the dosages of 2.50,5.00 and 10.00 μmol/L NaAsO2 (all P ＜ 0.05).Compared with the control group (0 h,100.00 ± 0.00),the levels of H3K36me3 modifications (48.47 ± 9.67,47.75 ± 6.98) were lower after treated with 10.00 μ mol/L NaAsO2 for 12 and 24 h (all P ＜ 0.05).③The levels of H3K36me3 modifications in HaCaT cells exposed to different doses of NaAsO2 were negatively associated with the tail DNA％ and OTM (r =-0.897,-0.903,all P ＜ 0.05).④Compared with the control group (0.00 μmol/L,100.00 ± 0.00),the mRNA levels of MGMT gene were lower at the dosages of 2.50,5.00 and 10.00 pmol/L NaAsO2 (78.20 ± 3.50,61.40 ± 2.60,49.15 ± 4.70,all P ＜ 0.05).⑤There was no observed H3K36me3 enrichmem regularity in the gene encoding ChIP1 and ChIP2 regions of MGMT gene in all doses of NaAsO2 groups (all P ＞ 0.05).Conclusions H3K36me3 may be involved in the regulation of arsenicinduced DNA damage in HaCaT cell.Amenic could inhibit the mRNA transcription of MGMT gene in HaCaT cells,but the transcription of MGMT gene regulate by H3K36me3 is not closely related to DNA damage induced by arsenic.