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Objective To investigate the role of histone H4 in the polarization of alveolar macrophages (AM) in lipopolysaccharide (LPS)-induced acute respiratory distress syndrome (ARDS) in mice. Methods i) The specific pathogen free male C57BL/6 mice were randomly divided into control group and 2, 4, 6 and 8 mg/kg LPS groups, with six mice in each group. The mice in the LPS groups were intratracheally administered LPS according to their respective doses, while the mice in the control group received an equivalent volume of 0.9% saline. After 12 hours, the arterial blood gas was analyzed, and the pulmonary edema and histopathological changes in lung tissues of mice in each group were observed. The level of histone H4 in bronchoalveolar lavage fluid (BALF) of mice was detected using enzyme-linked immunosorbent assay , and mice AMs of the five group were isolated using adherent method. ii) AMs from normal mice were isolated using adherent method and randomly divided into control group, histone H4 injury group, BALF injury group and anti-histone H4 antibody (anti-H4) intervention group. In the histone H4 injury group, AMs were treated with histone H4 at a final concentration of 20 mg/L. In the BALF injury group and anti-H4 intervention group, AMs were treated with 200 μL BALF supernatant from mice intratracheally administered 6 mg/kg body weight LPS, with the latter group treated with 25 mg/L anti-H4 antibody. The control group AMs were treated with phosphate-buffered saline. iii) After 12 hours of stimulation, the cells were collected, and the relative expression of tumor necrosis factor-α (Tnfa), interleukin-1β (Il1b), differentiation antigen 206 (Cd206) and arginase 1 (Arg1) in AMs was detected using real-time quantitative polymerase chain reaction. Results i) Compared with the control group, mice in all four LPS groups exhibited rapid breathing, inflammatory reaction and lung edema in lung tissues, which were aggravated in a dose-dependent manner. The ratio of partial pressure of arterial oxygen to fraction of inspired oxygen in mice decreased with the increase of LPS dose (P<0.05). The wet/dry weight ratio of lung, the level of histone H4 in BALF and the relative expression of Tnfa and Il1b mRNA in AMs increased with the increase of LPS dose (all P<0.05). The mice in the 6 and 8 mg/kg LPS groups developed ARDS. The level of histone H4 in BALF and the relative expression of Tnfa and Il1b mRNA in AMs of mice in 6 and 8 mg/kg LPS groups were higher than those in the other three groups (all P<0.05). ii) The relative expression of Tnfa and Il1b mRNA increased (both P<0.05), and the relative expression of Cd206 and Arg1 mRNA decreased (both P<0.05) in AMs of histone H4 injury group and BALF injury group compared with the control group. Compared with BALF injury group, the relative mRNA expression of Tnfa and Il1b in AMs of anti-H4 intervention group decreased (both P<0.05), while the relative expression of Arg1 mRNA increased (P<0.05). Conclusion LPS can induce a dose-dependent increase in histone H4 levels in BALF in mice. Histone H4 drives the development of ARDS by activating AMs to M1 polarization. Antagonizing histone H4 to interfere with AM polarization to M1 could be a target for the treatment of ARDS.
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{L-End}Objective To analyze the changes of seven potential biomarkers in plasma of patients with occupational silicosis (hereinafter referred to as "silicosis"), and explore their clinical value in determining the stage of silicosis. {L-End}Methods A total of 100 male silicosis patients were selected as the silicosis group (63 cases in stage Ⅰ and 37 cases in stage Ⅱ subgroups), and 100 male healthy individuals were selected as the control group using the 1∶1 matched case-control study. Enzyme-linked immunosorbent assay was used to analyze the level of interleukin-17 (IL-17), monocyte chemoattractant protein-1 (MCP-1), matrix metalloproteinase-9 (MMP-9), Krebs von den Lungen-6 (KL-6), connective tissue growth factor (CTGF), platelet-derived growth factor (PDGF), and histone H4 in plasma. Their clinical value for diagnosing silicosis was evaluated using receiver operating characteristic (ROC) curve, discriminant analysis stepwise method, and Fisher discriminant function analysis. {L-End}Results The levels of IL-17, MCP-1, MMP-9, KL-6, CTGF, PDGF, and histone H4 in the plasma of the silicosis group, silicosis stage Ⅰ subgroups, and stage Ⅱ subgroups were higher than those in the control group (all P<0.05). The levels of IL-17, MCP-1, and MMP-9 in the plasma of the stage Ⅱ subgroup decreased (all P<0.05), while the levels of KL-6, CTGF and histone H4 increased (all P<0.05) compared with the stage Ⅰ subgroup. The area under the ROC curve for diagnosing silicosis using these seven potential biomarkers ranged from 0.761 to 1.000 (all P<0.01), with the sensitivity of 0.640-1.000, the specificity of 0.840-0.990, and the Youden index of 0.540-0.990. The Fisher discriminant function was formed by stepwise discriminant analysis, and the results showed that the coincidence rate was 99.5%, and the misdiagnosis rate was 0.5% for diagnosing and staging silicosis with these seven potential biomarkers. The coincidence rate of diagnosing control group, silicosis stageⅠsubgroup and the silicosis stage Ⅱ subgroup was 100.0%, 98.4% and 100.0%, respectively. {L-End}Conclusion IL-17, MCP-1, MMP-9, KL-6, CTGF, PDGF and histone H4 in plasma can be used as biomarkers for the diagnosis of silicosis, and the Fisher discriminant function based on the combination of these seven biomarkers can assist in staging silicosis.
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Objective:To observe the changes of serum histone H4 level and its predictive value in patients with septic cardiomyopathy.Methods:A prospective study was conducted. A total of 147 patients with sepsis and septic shock were collected in emergency department. The general data were recorded. Transthoracic echocardiography and plasma histone H4 were conducted within 24 hours and 7 days after admission.The scores of sequential organ failure assessment(SOFA), acute physiology and chronic health evaluationⅡ(APACHEⅡ), and nutritional risk screening 2002 (NRS2002) were evaluated within 24 hours. According to whether septic cardiomyopathy occurred, the patients were divided into two groups, and dynamic changes of histone H4 on the first and seventh day of the two groups were observed. The factors influencing the occurrence of septic cardiomyopathy were analyzed by multivariate logistic regression. The prediction ability of serum histone H4 on septic cardiomyopathy was evaluated by receiver operating curve (ROC).Results:The incidence of septic cardiomyopathy was 28.6% (42 / 147). The level of histone H4 in septic cardiomyopathy group was higher than that in non septic cardiomyopathy group ( Z = 4.449, P < 0.001), and dynamic detection showed that the level of histone H4 on the seventh day was lower than that on admission ( Z=3.057, P=0.002). Multivariate logistic regression showed that the high serum histone H4 level [Odd Ratio( OR)=1.337, 95% confidence interval (95% CI) was 1.173-1.522, P < 0.001], SOFA ( OR= 1.474, 95% CI 1.227-1.769, P < 0.001), older age ( OR = 1.074, 95% CI 1.019-1.132, P = 0.008) were independent risk factors for septic cardiomyopathy. The area of ROC curve for serum histone H4 to predict septic cardiomyopathy was 0.729 ( P < 0.001), the predictive cut-off value was 10.81 ng/ml, which yielded a sensitivity 0.524 and a specificity of 0.914. Conclusions:The level of histone H4 showed dynamic change in septic cardiomyopathy, and high serum histone H4 level has a good predictive value for the occurrence of septic cardiomyopathy.
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Objective • To study whether lysine crotonylation can be used as a candidate biomarker for prostate cancer diagnosis, and its correlation with clinical stages and pathologic grades. Methods • Seventy-three cases of tumor and 7 normal prostate tissues were included in the study. The global levels of lysine crotonylation and histone H4 acetylation were detected in each sample by immunohistochemistry. Statistical comparison and correlation analysis were performed. Results • Compared with normal prostate tissue, the global level of lysine crotonylation was significantly reduced in prostate cancer tissue (P=0.001), while histone H4 acetylation levels were close to each other in two groups (P=0.704). No statistical difference in the levels of lysine crotonylation or histone H4 acetylation were found in different clinical stages and pathologic grades (P>0.05). There was no correlation between histone H4 acetylation and clinical stages or pathologic grades of prostate cancer. There was a positive correlation between lysine crotonylation and the grading of prostate cancer (r=0.493, P=0.000). Conclusion • Compared to histone H4 acetylation, lysine crotonylation is a better candidate biomarker to diagnose prostate cancer.
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Objective@#To evaluate the expression of histone H4 acetylation(Ac-H4) during the cleft palates formation induced by 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin(TCDD) in C57BL/6J mice.@*Methods@#Forty-eight pregnant C57BL/6J mice were completely randomly divided into two groups: ① TCDD group, mice were treated with 20ug/kg of TCDD on gestation day (GD) 10.5 by gastric perfusion; ② control group, mice were treated with an equivalent of corn oil. The head samples were collected and sliced in coronal plane on GD13.5, GD14.5 and GD15.5 respectively. Histone H4 acetylation in the palates were evaluated by immunohistochemical staining and Western Blot in the two groups.@*Results@#Histone H4 acetylation was mainly expressed in the palatal epithelial cells and slightly expressed in mesenchymal cells. The expression level of histone H4 acetylation was 0.6002±0.2530, 0.9180±0.0941 and 0.8966±0.0908 respectively in control group on GD13.5, GD14.5 and GD15.5; while 1.0229±0.2779, 1.6095±0.2651 and 1.2758±0.1251 in TCDD group. There were statistically significant differences between the control group and TCDD group (P<0.05).@*Conclusions@#The histone H4 acetylation was involved in the cleft palate formation induced by TCDD in C57BL/6J mice.
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Objective To study the effects of missense mutation of H4 Lysine 20,expore the molecular mechanisms,and find some specific interaction proteins,such as H4K20me3 demethyhransferase.Methods Stable HeLaS3 cell line expressed H4 or H4 mutations that fuses a Flag-HA tag to carboxyl(C)-terminal was established by virus infection.Mutations include lysine mutates to methionine and alanine.Harveste cells expressed purpose protein.Staining with propidium iodide (PI) and analyze cell cycle analysis by fluorescence activating cell sorter (FACS).Prepare mono nuclesome (mono-N) of three cells samples and purify associated protein complex by immunoprecipitation assay.Distingwish protein differences by Siliver staining and cut different protein bands for mass spectrometry analysis.Results HeLa S3 overexpressed H4K20M exhibit abnormal phenotype when transfected virus 72h,mainly show abnormal cell and nuclei volume increases,stop growing and no longer split.FACS shows cells overexpressed H4K20M were blocked in G2/M period.Mass spectrometry data shows the expression of PRBP4/7 increases in cells overexpressed H4K20M.Conclusion Histone H4 lysine 20 mutation to methionine block cell cycle,this block is likely to be caused by protein PRBP4/7.
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Objective To identify the differences of circulating histone H4 (hH4) levels in different phrase of children under 1 year old with sepsis, investigating its possible role in the process of sepsis. Methods We retrospectively analyzed 40 sepsis patients aged between 1 and 12 months old who were ad-mitted in PICU of Shenzhen Maternity and Children Healthcare Hospital between November 2016 and December 2017. Their clinical data were reviewed,and then they were categorized into three groups,non-criti-cal group(n=21),critical group(n=11) and extremely critical group(n=8) by Pediatric Critical Illness Score ( PCIS) . Forty children of the same age group with no signs of infection were enrolled as control group. Plasma from 4 groups on admission day and the third day after admission were collected. Plasma hH4 levels of all samples were measured and all parameters were reconfigured and statistically analyzed. Results HH4 levels of samples were collected from sepsis group on admission day:non-critical group (97. 22 ± 16. 92)μg/L,criti-cal group (148. 09 ± 33. 00)μg/L,extremely critical group (195. 04 ± 45. 85)μg/L. HH4 levels of all three sub groups were significantly higher than those of control group [(46. 07 ± 18. 06)μg/L, H = 64. 16, P<0. 001]. Data of the third day were as follow:non-critical group (98. 96 ± 16. 29)μg/L,critical group (152. 57 ± 29. 04)μg/L,extremely critical group (239. 52 ± 49. 84)μg/L. Comparing the parameters of the first and the third admission day,hH4 levels of both non-critical group (Z= -1. 42,P=0. 16) and critical group (Z= -1. 48,P=0. 14) showed no significant difference. However,hH4 levels of extremely critical group were significantly increased on the third admission day (Z= -2. 67,P=0. 008). Setting the discharge day or the seventh day after admission as the endpoint of the observation, hH4 levels of the death group [241. 53(229. 19,245. 07)μg/L]were significantly higher than that of the survival group[115. 21(96. 38,136. 15)μg/L,Z= -2. 80,P=0. 005]. Among the sepsis,hH4 levels were negatively correlated with their PCIS results on admission day(r= -0. 853,P<0. 001). The hH4 levels were also negatively correlated with the patients'PCIS score changes on the third day after admission(r = -0. 554,P <0. 001). Conclusion The levels of circulating hH4 in sepsis children are co-related to the prognosis and severity suggesting that it may affect the process of the sepsis in certain ways.
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BACKGROUND: Epigenetic alteration may affect a patient's prognosis by altering the development and progression of the tumor. Some recent reports have identified a correlation between histone modification and patient outcome. However, no studies have been conducted on global histone modification in osteosarcomas. METHODS: We investigated histone modification in 54 cases of osteosarcoma by performing immunohistochemical staining. The immunohistochemical expression of four histone modification markers, acetylated H4 lysine 12 (H4K12Ac), acetylated H3 lysine 18, trimethylated H3 lysine 27, and dimethylated H3 lysine 4 were evaluated. RESULTS: High H4K12Ac expression was correlated with patient age (p=0.011). However, the other histone modification markers showed no correlation with any of the clinicopathological data such as survival, tumor grade, tumor site, metastasis, age, or gender. CONCLUSIONS: Our study showed that all four histone modification markers are expressed in osteosarcoma (median expression rate, 40 to 60%). However, we did not find a correlation with the clinicopathological factors except for age. Further study to evaluate the reason for the association between H4K12Ac and patient age is needed.