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Background. Systemic lupus erythematosus (SLE) is characterized by poor regulation of the immune response leading to chronic inflammation and multiple organ dysfunction. Glucocorticoid (GC) is currently one of the main treatments. However, a high dose or prolonged use of GC may result in glucocorticoid-induced osteoporosis (GIOP). Jiedu Quyu Ziyin decoction (JP) is effective in treating SLE and previous clinical studies have proved that JP can prevent and treat SLE steroid osteoporosis (SLE-GIOP). We aim to examine JPs main mechanism on SLE-GIOP through network pharmacology and molecular docking. Methods. TCMSP and TCMID databases were used to screen potential active compounds and targets of JP. The SLE-GIOP targets are collected from GeneCards, OMIM, PharmGkb, TTD, and DrugBank databases. R software was used to obtain the cross-targets of JP and SLE-GIOP and to perform GO and KEGG enrichment analysis. Cytoscape software was used to make the Chinese Medicines-Active Ingredient-Intersection Targets network diagram. STRING database construct protein–protein interaction network and obtain the core targets. Auto Dock Tools and Pymol software were used for docking. Results. Fifty eight targets overlapped between JP and SLE-GIOP were suggested as potential targets of JP in the treatment of SLE-GIOP. Network topology analysis identified five core targets. GO enrichment analysis was obtained 1,968 items, and the top 10 biological process, closeness centrality, and molecular function were displayed. A total of 154 signaling pathways were obtained by KEGG enrichment analysis, and the top 30 signaling pathways were displayed. JP was well bound by MAPK1, TP53, and MYC according to the molecular docking results. Conclusion. We investigated the potential targets and signaling pathways of JP against SLE-GIOP in this study. It shows that JP is most likely to achieve the purpose of treating SLE-GIOP by promoting the proliferation and differentiation of osteoblasts. A solid theoretical foundation will be provided for the future study of clinical and experimental topics.

Supplementary Figure S1. Decreased survival rate of lung cancer patients who are older than 65 and with high STAT3 activation in TAMs Supplementary Figure S2. STAT3 knockout in myeloid cells in STAT3DMye mice Supplementary Figure S3. STAT3 knockout in myeloid cells in STAT3DMye mice Supplementary Figure S4. Decreased lung tumorigenesis in urethane-treated STAT3DMye mice Supplementary Figure S5. Same pathology of the lung tumors in WT and STAT3DMye mice treated with urethane Supplementary Figure S6. Different changes in the total numbers of lung macrophages, neutrophils and lymphocytes in urethane-treated STAT3DMyc mice Supplementary Figure S7. Increased M1 macrophages but decreased M2 macrophages in the lungs of urethane-treated STAT3DMye mice Supplementary Figure S8. Lung myeloid and lymphoid cell changes in STAT3DMye mice treated with urethane Supplementary Figure S9. Expression changes of chemokines in mouse lungs by urethane Supplementary Figure S10. T-cell apoptosis by MDSCs Supplementary Figure S11. Defect in T-cell apoptosis induction by STAT3 KO macrophages Supplementary Figure S12. No obvious effect on T-cell proliferation and lung recruitment by myeloid STAT3 deletion Supplementary Figure S13. Treg cell differentiation by WT macrophages but not by STAT3 KO macrophages Supplementary Figure S14. Reversal of the decreased lung tumorigenesis in urethane-treated STAT3DMye mice by CD8 deletion Supplementary Figure S15. Quantification of the iNOS and arginase staining in the Figure 7F Supplementary Figure S16. Reduced STAT3 activation in lung tumor cells in STAT3DMye mice Supplementary Table S1. Antibodies used in this study Supplementary Table S2. Clinicopathological characteristics of lung cancer patients Supplementary Table S3. Primer pairs used for real-time PCR in the study

One of the most fundamental and challenging questions in the cancer field is how immunity in patients with cancer is transformed from tumor immunosurveillance to tumor-promoting inflammation. Here, we identify the transcription factor STAT3 as the culprit responsible for this pathogenic event in lung cancer development. We found that antitumor type 1 CD4+ T-helper (Th1) cells and CD8+ T cells were directly counter balanced in lung cancer development with tumor-promoting myeloid-derived suppressor cells (MDSCs) and suppressive macrophages, and that activation of STAT3 in MDSCs and macrophages promoted tumorigenesis through pulmonary recruitment and increased resistance of suppressive cells to CD8+ T cells, enhancement of cytotoxicity toward CD4+ and CD8+ T cells, induction of regulatory T cell (Treg), inhibition of dendritic cells (DC), and polarization of macrophages toward the M2 phenotype. The deletion of myeloid STAT3 boosted antitumor immunity and suppressed lung tumorigenesis. These findings increase our understanding of immune programming in lung tumorigenesis and provide a mechanistic basis for developing STAT3-based immunotherapy against this and other solid tumors. Cancer Immunol Res; 5(3); 257–68. ©2017 AACR.

P-TEFb modulates RNA polymerase II elongation through alternative interaction with negative and positive regulation factors. While inactive P-TEFbs are mainly sequestered in the 7SK snRNP complex in a chromatin-free state, most of its active forms are in complex with its recruitment factors, Brd4 and SEC, in a chromatin-associated state. Thus, switching from inactive 7SK snRNP to active P-TEFb (Brd4/P-TEFb or SEC/P-TEFb) is essential for global gene expression. Although it has been shown that cellular signaling stimulates the disruption of 7SK snRNP, releasing dephosphorylated and catalytically inactive P-TEFb, little is known about how the inactive released P-TEFb is reactivated. Here, we show that the Cdk9/CycT1 heterodimer released from 7SK snRNP is completely dissociated into monomers in response to stress. Brd4 or SEC then recruits monomerized Cdk9 and CycT1 to reassemble the core P-TEFb. Meanwhile, the binding of monomeric dephosphorylated Cdk9 to either Brd4 or SEC induces the autophosphorylation of T186 of Cdk9. Finally, the same mechanism is employed during nocodazole released entry into early G1 phase of cell cycle. Therefore, our studies demonstrate a novel mechanism by which Cdk9 and CycT1 monomers are reassembled on chromatin to form active P-TEFb by its interaction with Brd4 or SEC to regulate transcription.

Compared to conventional sheet forming operations, incremental sheet forming (ISF) is a flexible forming technique that can achieve higher formability in terms of localized deformation. Due to excellent mechanical properties and X-ray penetration, polyether-ether-ketone (PEEK) is an ideal alternative to titanium alloy and stainless steel in orthopedic applications. In this study, a 3-axis desktop manufacturing system has been fabricated to investigate the temperature-dependent formability of PEEK in terms of manufacturing the cranial plate by using the ISF technique. Meanwhile, the forming force, temperature distribution, geometrical accuracy, and thermal properties were obtained and analyzed. The findings indicate that the ISF technique provides technological and economic advantages in cranial reconstruction by using PEEK.

Posttranslational modifications of NF-κB, including phosphorylation, acetylation, and methylation, have emerged as important regulatory mechanisms to control the transcriptional outcomes of this important transcription factor. These modifications work independently, sequentially or in combination to modulate the diverse biological functions of NF-κB in cancer and inflammatory response. Here, we describe some experimental methods to detect the in vitro and in vivo phosphorylation and acetylation of NF-κB, specifically focusing on the RelA subunit of NF-κB. These methods include labeling the phospho- or acetyl- groups with radioisotopes in vitro and immunoblotting with site-specific anti-phospho-serine or acetyl-lysine antibodies in culture cells and tissue samples.

Macrophage-mediated inflammatory response has been implicated in the pathogenesis of obesity and insulin resistance. Brd4 has emerged as a key regulator in the innate immune response. However, the role of Brd4 in obesity-associated inflammation and insulin resistance remains uncharacterized. Here, we demonstrated that myeloid lineage-specific Brd4 knockout (Brd4-CKO) mice were protected from high-fat diet-induced (HFD-induced) obesity with less fat accumulation, higher energy expenditure, and increased lipolysis in adipose tissue. Brd4-CKO mice fed a HFD also displayed reduced local and systemic inflammation with improved insulin sensitivity. RNA-Seq of adipose tissue macrophages (ATMs) from HFD-fed WT and Brd4-CKO mice revealed that expression of antilipolytic factor Gdf3 was significantly decreased in ATMs of Brd4-CKO mice. We also found that Brd4 bound to the promoter and enhancers of Gdf3 to facilitate PPARγ-dependent Gdf3 expression in macrophages. Furthermore, Brd4-mediated expression of Gdf3 acted as a paracrine signal targeting adipocytes to suppress the expression of lipases and the associated lipolysis in cultured cells and mice. Controlling the expression of Gdf3 in ATMs could be one of the mechanisms by which Brd4 modulates lipid metabolism and diet-induced obesity. This study suggests that Brd4 could be a potential therapeutic target for obesity and insulin resistance.

Dong et al. show that the epigenetic regulator Brd4 forms a complex with transcription factors IRF8/PU.1 to activate the transcription of NLRC4 inflammasome component Naips in macrophages. Brd4-dependent inflammasome activation protects mice from Salmonella infection via cytokine release and pyroptosis., NLRC4 inflammasome activation and the subsequent maturation of IL-1β and IL-18 are critical for protection against infection by bacterial pathogens. The epigenetic regulator Brd4 has emerged as a key player in inflammation by regulating the expression of inflammatory cytokines. However, whether Brd4 has any role in inflammasome activation remains undetermined. Here, we demonstrated that Brd4 is an important regulator of NLRC4 inflammasome activation in response to Salmonella typhimurium infection. Brd4-deficient bone marrow–derived macrophages (BMDMs) displayed impaired caspase-1 activation, ASC oligomerization, IL-1β maturation, gasdermin-D cleavage, and pyroptosis in response to S. typhimurium infection. RNA sequencing and RT-PCR results revealed that the transcription of Naips was decreased in Brd4-deficient BMDMs. Brd4 formed a complex with IRF8/PU.1 and bound to the IRF8 and PU.1 binding motifs on the promoters of Naips to maintain the expression of Naips. Furthermore, myeloid lineage–specific Brd4 conditional knockout mice were more susceptible to S. typhimurium infection with increased mortality, bacterial loads, and tissue damage; impaired inflammasome-dependent cytokine production; and pyroptosis. Our studies identify a novel function of Brd4 in innate immunity by controlling inflammasome-mediated cytokine release and pyroptosis to effectively battle S. typhimurium infection.

Posttranslational modifications of NF-κB, including phosphorylation, acetylation, and methylation, have emerged as important regulatory mechanisms to control the transcriptional outcomes of this important transcription factor. These modifications work independently, sequentially or in combination to modulate the diverse biological functions of NF-κB in cancer and inflammatory response. Here, we describe some experimental methods to detect the in vitro and in vivo phosphorylation and acetylation of NF-κB, specifically focusing on the RelA subunit of NF-κB. These methods include labeling the phospho- or acetyl- groups with radioisotopes in vitro and immunoblotting with site-specific anti-phospho-serine or acetyl-lysine antibodies in culture cells and tissue samples.

H. pylori infection is one of the leading causes of gastric cancer and the pathogenicity of H. pylori infection is associated with its ability to induce chronic inflammation and apoptosis resistance. While H. pylori infection-induced expression of pro-inflammatory cytokines for chronic inflammation is well studied, the molecular mechanism underlying the apoptosis resistance in infected cells is not well understood. In this study, we demonstrated that H. pylori infection-induced apoptosis resistance in gastric epithelial cells triggered by Raptinal, a drug that directly activates caspase-3. This resistance resulted from the induction of cIAP2 (encoded by BIRC3) since depletion of BIRC3 by siRNA or inhibition of cIAP2 via BV6 reversed H. pylori-suppressed caspase-3 activation. The induction of cIAP2 was regulated by H. pylori-induced BIRC3 eRNA synthesis. Depletion of BIRC3 eRNA decreased H. pylori-induced cIAP2 and reversed H. pylori-suppressed caspase-3 activation. Mechanistically, H. pylori stimulated the recruitment of bromodomain-containing factor Brd4 to the enhancer of BIRC3 and promoted BIRC3 eRNA and mRNA synthesis. Inhibition of Brd4 diminished the expression of BIRC3 eRNA and the anti-apoptotic response to H. pylori infection. Importantly, H. pylori isogenic cagA-deficient mutant failed to activate the synthesis of BIRC3 eRNA and the associated apoptosis resistance. Finally, in primary human gastric epithelial cells, H. pylori also induced resistance to Raptinal-triggered caspase-3 activation by activating the Brd4-dependent BIRC3 eRNA synthesis in a CagA-dependent manner. These results identify a novel function of Brd4 in H. pylori-mediated apoptosis resistance via activating BIRC3 eRNA synthesis, suggesting that Brd4 could be a potential therapeutic target for H. pylori-induced gastric cancer.

Activation of farnesoid X receptor (FXR) by obeticholic acid (OCA) reduces hepatic inflammation and fibrosis in patients with primary biliary cholangitis (PBC), a life-threatening cholestatic liver failure. Inhibition of bromodomain-containing protein 4 (BRD4) also has antiinflammatory, antifibrotic effects in mice. We determined the role of BRD4 in FXR function in bile acid (BA) regulation and examined whether the known beneficial effects of OCA are enhanced by inhibiting BRD4 in cholestatic mice. Liver-specific downregulation of BRD4 disrupted BA homeostasis in mice, and FXR-mediated regulation of BA-related genes, including small heterodimer partner and cholesterol 7 alpha-hydroxylase, was BRD4 dependent. In cholestatic mice, JQ1 or OCA treatment ameliorated hepatotoxicity, inflammation, and fibrosis, but surprisingly, was antagonistic in combination. Mechanistically, OCA increased binding of FXR, and the corepressor silencing mediator of retinoid and thyroid hormone receptor (SMRT) decreased NF-κB binding at inflammatory genes and repressed the genes in a BRD4-dependent manner. In patients with PBC, hepatic expression of FXR and BRD4 was significantly reduced. In conclusion, BRD4 is a potentially novel cofactor of FXR for maintaining BA homeostasis and hepatoprotection. Although BRD4 promotes hepatic inflammation and fibrosis in cholestasis, paradoxically, BRD4 is required for the antiinflammatory, antifibrotic actions of OCA-activated FXR. Cotreatment with OCA and JQ1, individually beneficial, may be antagonistic in treatment of liver disease patients with inflammation and fibrosis complications., Treatment with obeticholic acid, a farnesoid X receptor agonist, or JQ1, a bromodomain-containing protein 4 inhibitor, has antiinflammatory, antifibrotic effects in cholestatic mice, but beneficial effects are lost in combination.

To investigate the related factors influencing plasma transfusion efficacy so as to improve the plasma transfusion efficiency.According to the clinical symptoms and the laboratorial results, the patients were divided into transfusion efficient and inefficient groups. A total of13090.8 units of plasma were transfused to 4423 patients. The clinical symptoms and the hemorrhage related index per- and pro-transfusion, plasma components sorts, storage time, and the dose of plasma (kg/ml) transfusion were analyzed.The largest transfusion volume of plasma were in intensive care unit (ICU) accounted for 30.36%, the largest blood plasma per patient transfusion was in cardiac surgery (3.96 U). The analysis of transfusion efficiency showed that in terms of patient age, there were difference in transfusion efficiency among the patients with different ages (P＜0.001). The effective transfusion rate in the group of age ＜18 was 53%, which was higher than that in group of age 18-60(41%) and group of age ＞60 (30%); in terms of sex, the effective transfusion rate in female group was higher than that in male group (42% vs 37%) (P＜0.001); in terms of transfusion plasma volume/body weight, there were differences in transfusion efficiency (P＞0.05). The multi-factor logistic regression analysis showed that there was no significant correlation among the plasma sorts, storage time of the plasma pre-transfusion and transfusion efficiency(P＞0.05). The analysis of the non-hemolytic fever reaction caused by plasma transfusion revealed that there was no statistical difference between the plasma and the leukocyte-depleted plasma groups (P＞0.05).The plasma transfusion effectiveness relates with age and sex, but not relates with the transfusion plasma voume/body weight, plasma sorts, and the duration of storage.4 423例次血浆输注效果的影响因素分析研究.探讨影响血浆输注效果的相关因素，以期进一步提高患者血浆输注有效率和临床输注的合理性.根据输血后患者临床症状改善情况和实验室凝血检测值将患者分为血浆输注有效和无效2组。对4 423例患者的基本临床资料和13 090.8 U的血浆输注过程中涉及的主要因素如输血浆前PT、APTT、血浆种类、保存时间、输注血浆体重比（kg/ml）等进行分类统计分析.外科监护室患者血浆用量最多，占总血浆输注比例最大（30.36%），人均用血浆量最大的科室为心外科（3.96 U）。从输注效果上看，不同年龄组患者的输血效果有差别（P＜0.001），＜18岁组患者血浆输注有效率（53%）高于18-60岁和＜60岁组，18-60岁组和＞60岁组患者血浆输注有效率分别为41%和30%；女性患者输血浆有效率（42%）高于男性（37%）（P＜0.001）；按不同血浆体重比进行血浆输注效果比较显示无显著差异（P＞0.05）；通过多因素Logistic回归分析，普通冰冻血浆、去白细胞血浆和新鲜冰冻血浆3类血浆的临床输注效果间没有差异；输注前血浆保存时间与输注效果之间无显著相关性。去白细胞血浆与未经去白的血浆相比，并未降低非溶血性发热性输血反应的发生率.血浆输注效果的主要影响因素有患者年龄、性别，但与血浆体重比、血浆种类和血浆保存时间关系不大；只要于保存期内输注，保存时间的长短与输注效果间没有显著相关性.

The BWH (broadband excitation receiving, wide azimuth observation, high density acquisition) seismic data size deployed in the deep complex tectonic area of the northern Songliao Basin was 7.5T, and the surface elevation of the study area varies greatly. The single point receiving observing system with high density and small cells presents a new challenge to static processing. The pick-up precision of the first break wave directly determines whether the static correction is accurate or not. The conventional method can pick up the first break wave in the shot domain, and the precision of the pick-up quantity cannot be required by the static correction of tomography. In this paper, according to the characteristics of BHW mass data, a new method is proposed to separate the data into small pick-up units by using detection lines as picking units and dividing offset groups into small pick-up units. The trend line which controls the pick-up accuracy is adapted to the influence of the far and near offset at the same time, and the pick-up accuracy is greatly improved. The static correction effect is achieved by the method of model-constrained travel-time tomography static correction. This method is especially suitable for BWH seismic data and other big offset data, the first break wave pick-up of high-density 3D seismic data received by single detect point can improve the pick-up accuracy to the maximum extent, and has wide application and popularization space.

Numerical simulations of turbulent flows in a stirred dead-end membrane bioreactor are performed by using the RNG k - e model based on the finite volume method with the software Fluent. Comparisons of numerical and experimental results confirm the reliability and the feasibility of the constructed model. The flow structures such as the wake flows and the circulation loops in the stirred flows are well simulated. An increase of stirring speed is proposed to minimize the low velocity region. The single vane stirrer is found to be beneficial for biological separations. Results suggest that the increase of the vane number can enhance the mixing effect in the flow domains. However, a circular disk stirrer goes against

Pin1 is a peptidyl-prolyl cis-trans isomerase that specifically binds to a phosphorylated serine or threonine residue preceding a proline (pSer/Thr-Pro) motif and catalyzes the cis-trans isomerization of proline imidic peptide bond, resulting in conformational change of its substrates. Pin1 regulates many biological processes and is also involved in the development of human diseases, like cancer and neurological diseases. Many Pin1 substrates are transcription factors and transcription regulators, including RNA polymerase II (RNAPII) and factors associated with transcription initiation, elongation, termination and post-transcription mRNA decay. By changing the stability, subcellular localization, protein-protein or protein-DNA/RNA interactions of these transcription related proteins, Pin1 modulates the transcription of many genes related to cell proliferation, differentiation, apoptosis and immune response. Here, we will discuss how Pin regulates the properties of these transcription relevant factors for effective gene expression and how Pin1-mediated transcription contributes to the diverse pathophysiological functions of Pin1.

Metabolic syndrome (MetS) has been shown to be associated with an increased risk of gastric cancer. However, the impact of MetS on gastric cancer mortality remains largely unknown. Here, we prospectively examined the prediction of preoperative MetS for gastric cancer mortality by analyzing a subset of data from the ongoing Fujian prospective investigation of cancer (FIESTA) study. This study was conducted among 3012 patients with gastric cancer who received radical gastrectomy between 2000 and 2010. The latest follow-up was completed in 2015. Blood/tissue specimens, demographic and clinicopathologic characteristics were collected at baseline. During 15-year follow-up, 1331 of 3012 patients died of gastric cancer. The median survival time (MST) of patients with MetS was 31.3 months, which was significantly shorter than that of MetS-free patients (157.1 months). The coexistence of MetS before surgery was associated with a 2.3-fold increased risk for gastric cancer mortality (P, Highlights • We analyzed the impact of preoperative metabolic syndrome on the survival of 3012 gastric cancer patients after surgery over 15-year follow-up. • This is by far the largest cohort study to evaluate the association between the preoperative metabolic syndrome and gastric cancer mortality. • Preoperative metabolic syndrome, especially hyperglycemia, was predictive of significant gastric cancer mortality in patients after surgery. On the basis of 3012 patients with gastric cancer from the FIESTA study, we analyzed the impact of the preoperative MetS status on the survival of patients after radical gastrectomy over a period of 15-year follow-up. We found that preoperative MetS, especially hyperglycemia, was predictive of significant gastric cancer mortality in patients with radical gastrectomy, especially for early stage of gastric cancer. Pending its future validation in other large cohort studies, our findings could provide the basis for future personalized medicine, namely, gastric cancer patients with preoperative MetS should be identified early and treated with optimal regimens since these patients could have a poor survival probability.

The innate immune sensor NLRP3 assembles an inflammasome complex with NEK7 and ASC to activate caspase‐1 and drive the maturation of proinflammatory cytokines IL‐1β and IL‐18. NLRP3 inflammasome activity must be tightly controlled, as its over‐activation is involved in the pathogenesis of inflammatory diseases. Here, we show that NLRP3 inflammasome activation is suppressed by a centrosomal protein Spata2. Spata2 deficiency enhances NLRP3 inflammasome activity both in the macrophages and in an animal model of peritonitis. Mechanistically, Spata2 recruits the deubiquitinase CYLD to the centrosome for deubiquitination of polo‐like kinase 4 (PLK4), the master regulator of centrosome duplication. Deubiquitination of PLK4 facilitates its binding to and phosphorylation of NEK7 at Ser204. NEK7 phosphorylation in turn attenuates NEK7 and NLRP3 interaction, which is required for NLRP3 inflammasome activation. Pharmacological or shRNA‐mediated inhibition of PLK4, or mutation of the NEK7 Ser204 phosphorylation site, augments NEK7 interaction with NLRP3 and causes increased NLRP3 inflammasome activation. Our study unravels a novel centrosomal regulatory pathway of inflammasome activation and may provide new therapeutic targets for the treatment of NLRP3‐associated inflammatory diseases.

This paper studied the inhibitory effect of pomegranate peel (PP) extract on the growth of Microcystis aeruginosa, the model of harmful algal blooms in aquatic environment. The allelochemicals were identified by HPLC–MS/MS from PP and tested by batch experiment through measurement of algal density, chlorophyll a (Chl-a) concentration, maximum quantum yield of photosystem II (Fv/Fm), superoxide dismutase (SOD), and malondialdehyde (MDA) contents. Results showed that both PP powder and PP extract had obvious inhibitory effect on M. aeruginosa growth. Quercetin and luteolin were identified as the allelochemicals to M. aeruginosa growth. However, the inhibitory capacity of luteolin was stronger than that of quercetin. The growth inhibition ratio of luteolin can reach up to 98.7 and 99.1% of the control on day 7 at the dosages of 7 and 10 mg/L, respectively. Moreover, the changes of Chl-a, Fv/Fm, SOD, and MDA in M. aeruginosa confirmed jointly that the allelochemicals cause inhibition of photosystem and oxidative damage to M. aeruginosa cells with the antioxidant defense system being activated, which leads to the aggravation of membrane lipid peroxidation. Thus, luteolin could be used as a promising algaecide for emergency handling of M. aeruginosa blooms. This study might provide a new direction in the management of eutrophication in the future.

The association of DSIF and NELF with initiated RNA Polymerase II (Pol II) is the general mechanism for inducing promoter-proximal pausing of Pol II. However, it remains largely unclear how the paused Pol II is released in response to stimulation. Here, we show that the release of the paused Pol II is cooperatively regulated by multiple P-TEFbs which are recruited by bromodomain-containing protein Brd4 and super elongation complex (SEC) via different recruitment mechanisms. Upon stimulation, Brd4 recruits P-TEFb to Spt5/DSIF via a recruitment pathway consisting of Med1, Med23 and Tat-SF1, whereas SEC recruits P-TEFb to NELF-A and NELF-E via Paf1c and Med26, respectively. P-TEFb-mediated phosphorylation of Spt5, NELF-A and NELF-E results in the dissociation of NELF from Pol II, thereby transiting transcription from pausing to elongation. Additionally, we demonstrate that P-TEFb-mediated Ser2 phosphorylation of Pol II is dispensable for pause release. Therefore, our studies reveal a co-regulatory mechanism of Brd4 and SEC in modulating the transcriptional pause release by recruiting multiple P-TEFbs via a Mediator- and Paf1c-coordinated recruitment network.

This study investigated how efficiently CaO2 could treat black-odor landscape water caused by low dissolved oxygen (DO) in a field experiment of 600 m2. The study demonstrated that CaO2 could significantly elevate the DO concentration in waters and the oxidation–reduction potential (ORP) level in sediments (p = 0.003 and p = 0), which is conducive to improving the anoxic environment of landscape water. The concentrations of total chemical oxygen demand (TCOD) and S2− in overlying and interstitial waters were considerably decreased. The average concentrations of TCOD in the overlying and interstitial waters of the test zone (TZ) were 52.98% and 66.05% of those of the control zone (CZ), and the average concentrations of S2− in the overlying and interstitial waters of TZ were 29.63% and 39.79% of those of CZ. Meanwhile, CaO2 could obviously reduce turbidity but increase the transparency in the overlying water. The mean value of turbidity in the overlying water of TZ was 39.46% of that of CZ, whereas the transparency in the overlying water of TZ was 2.07 times that of CZ. Furthermore, CaO2 changed the microbial community structure in the sediments, where the relative abundance of anaerobic bacteria was decreased but that of the aerobic bacteria was increased with some functional bacteria. In summary, CaO2 could significantly increase the DO and ORP in black-odor landscape water, obviously inhibit the release of pollutants from sediment, and increase the diversity of microbial strains. Consequently, the black-odor phenomenon of landscape water could be alleviated effectively by adding CaO2.

Inhibition of excessive osteoclast differentiation and activity is a valid approach for the treatment of osteoporosis. T63 is a small-molecule compound identified from a high throughput screening based on RUNX2 transcriptional activity, and has been reported to stimulate osteoblast formation. However, whether the compound has any effect on osteoclast differentiation remains unknown. Here, we examined the in vitro effect of T63 on osteoclastogenesis. T63 was found to inhibit the number of TRAP-positive cells in an osteoblast-osteoclast co-culture system, and inhibited Rankl expression in the preosteoblast MC3T3-E1 cells. The compound also directly suppressed RANKL-induced osteoclast differentiation in both dose- and time-dependent manner, as evidenced by the decrease of TRAP activity, F-actin formation and osteoclastogenesis-related genes expression in RAW264.7 cells. Moreover, pretreatment with T63 markedly decreased the activation of mitogen-activated protein kinases and Akt, both of which are positively involved in the regulation of osteoclastogenesis. Collectively, our findings suggest T63 has a protective effect against bone loss by inhibiting bone resorption. Its regulatory effect on bone metabolism makes the compound a more promising candidate for the potential application in the treatment of osteoporosis.

Current clinical treatment of Helicobacter pylori infection, the main etiological factor in the development of gastritis, gastric ulcers, and gastric carcinoma, requires a combination of at least two antibiotics and one proton pump inhibitor. However, such triple therapy suffers from progressively decreased therapeutic efficacy due to the drug resistance and undesired killing of the commensal bacteria due to poor selectivity. Here, we report the development of antimicrobial polypeptide-based monotherapy, which can specifically kill H. pylori under acidic pH in the stomach while inducing minimal toxicity to commensal bacteria under physiological pH. Specifically, we designed a class of pH-sensitive, helix–coil conformation transitionable antimicrobial polypeptides (HCT-AMPs) (PGA) m - r -(PHLG-MHH) n , bearing randomly distributed negatively charged glutamic acid and positively charged poly(γ-6- N -(methyldihexylammonium)hexyl-l-glutamate) (PHLG-MHH) residues. The HCT-AMPs showed unappreciable toxicity at physiological pH when they adopted random coiled conformation. Under acidic condition in the stomach, they transformed to the helical structure and exhibited potent antibacterial activity against H. pylori , including clinically isolated drug-resistant strains. After oral gavage, the HCT-AMPs afforded comparable H. pylori killing efficacy to the triple-therapy approach while inducing minimal toxicity against normal tissues and commensal bacteria, in comparison with the remarkable killing of commensal bacteria by 65% and 86% in the ileal contents and feces, respectively, following triple therapy. This strategy renders an effective approach to specifically target and kill H. pylori in the stomach while not harming the commensal bacteria/normal tissues.

Osteoporosis results from the imbalance between bone resorption and bone formation, and restoring the normal balance of bone remodeling is highly desirable for identification of better treatment. In this study, using a cell-based high-throughput screening model representing Runt-related transcription factor 2 (RUNX2) transcriptional activity, we identified a novel small-molecular-weight compound, T63, as an efficient up-regulator of osteogenesis. T63 increased the alkaline phosphatase (ALPL) activity and mineralization as well as gene expression of Alpl and other osteogenic marker genes in mouse osteoblasts and mesenchymal stem cell-like cells. Upon induction of osteoblast differentiation, T63 inhibited adipogenic differentiation in the pluripotent mesenchymal cells. Consistently, T63 up-regulated RUNX2 mRNA and protein levels, and knockdown of RUNX2 reduced the osteogenic role of T63. Mechanistically, T63 activated both BMPs and WNT/β-catenin signaling pathways. Inhibition of either signaling pathway with specific inhibitor suppressed T63-induced RUNX2 expression and the osteogenic phenotypes. Moreover, T63 markedly protected against bone mass loss in the ovariectomized and dexamethasone treated rat osteoporosis model. Collectively, our data demonstrate that T63 could be a promising drug candidate and deserves further development for potential therapeutics in osteoporosis.

Bromodomain-containing factor Brd4 has emerged as an important transcriptional regulator of NF-κB-dependent inflammatory gene expression. However, the in vivo physiological function of Brd4 in the inflammatory response remains poorly defined. We now demonstrate that mice deficient for Brd4 in myeloid-lineage cells are resistant to LPS-induced sepsis but are more susceptible to bacterial infection. Gene-expression microarray analysis of bone marrow-derived macrophages (BMDMs) reveals that deletion of Brd4 decreases the expression of a significant amount of LPS-induced inflammatory genes while reversing the expression of a small subset of LPS-suppressed genes, including MAP kinase-interacting serine/threonine-protein kinase 2 (Mknk2). Brd4-deficient BMDMs display enhanced Mnk2 expression and the corresponding eukaryotic translation initiation factor 4E (eIF4E) activation after LPS stimulation, leading to an increased translation of IκBα mRNA in polysomes. The enhanced newly synthesized IκBα reduced the binding of NF-κB to the promoters of inflammatory genes, resulting in reduced inflammatory gene expression and cytokine production. By modulating the translation of IκBα via the Mnk2-eIF4E pathway, Brd4 provides an additional layer of control for NF-κB-dependent inflammatory gene expression and inflammatory response.

Aesculin (AES), a coumarin compound derived from Aesculus hippocasanum L, is reported to exert protective role against inflammatory diseases, gastric disease and cancer. However, direct effect of AES in bone metabolism is deficient. In this study, we examined the effects of AES on osteoclast (OC) differentiation in receptor activator of NF-κB ligand (RANKL)-induced RAW264.7 cells. AES inhibits the OC differentiation in both dose- and time-dependent manner within non-toxic concentrations, as analyzed by Tartrate Resistant Acid Phosphatase (TRAP) staining. The actin ring formation manifesting OC function is also decreased by AES. Moreover, expressions of osteoclastogenesis related genes Trap, Atp6v0d2, Cathepsin K and Mmp-9 are decreased upon AES treatment. Mechanistically, AES attenuates the activation of MAPKs and NF-κB activity upon RANKL induction, thus leading to the reduction of Nfatc1 mRNA expression. Moreover, AES inhibits Rank expression, and RANK overexpression markedly decreases AES's effect on OC differentiation and NF-κB activity. Consistently, AES protects against bone mass loss in the ovariectomized and dexamethasone treated rat osteoporosis model. Taken together, our data demonstrate that AES can modulate bone metabolism by suppressing osteoclastogenesis and related transduction signals. AES therefore could be a promising agent for the treatment of osteoporosis.

Background: Increased occurrence of chronic syndromes has prompted researchers to investigate and develop drugs and methods for controlling chronic syndromes with a view to improve human health and reduce early aging. Material and methods: Human trials: After the allotted multivitamin pills or placebo pills had been taken for a stipulated period of about 2 months, the volunteers filled out feedback forms on curative effects of the pills in line with the health examination reports. The effects of the multivitamin on various symptoms or diseases and dysfunctions of the chronic metabolic syndromes were noted and evaluated based on the information provided in forms. Animal experiments: Mouse aging model induced by D-galactose were administered the multivitamin by oral gavage every morning. At the end of the sixth week, activity or content of the components associated with ageing and anti-aging in the brain and liver of the aging mice were determined to investigate the mechanisms of the new multivitamin on chronic metabolic syndromes and aging. Results: We found that multivitamin can eliminate or attenuate 38 types of symptoms or dysfunctions of the investigated metabolic syndromes; and that it has both preventive and curative/adjunctive therapeutic effects on the metabolic syndromes. The effects of this multivitamin on components associated with aging and anti-aging were significantly decreased - malondialdehyde content and monoamine oxidase activity but significantly increased activity of superoxide dismutase and glutathione peroxidase. This multivitamin has significant anti-aging effects. Conclusion: Supplementing with this multivitamin can prevent and provide treatment/adjunctive therapy for these chronic metabolic syndromes and delay the aging process. List of Abbreviations BW body weight; Cu/Zn-SOD, cuprum/zinc-superoxide dismutase MAO monoamine oxidase MDA malondialdehyde; Mn-SOD, manganese-superoxide dismutase; T-SOD, total superoxide dismutase; TP, total protein

Acetylation of the RelA subunit of NF-κB at lysine-310 regulates the transcriptional activation of NF-κB target genes and contributes to maintaining constitutively active NF-κB in tumors. Bromodomain-containing factor Brd4 has been shown to bind to acetylated lysine-310 (AcLys310) and to regulate the transcriptional activity of NF-κB, but the role of this binding in maintaining constitutively active NF-κB in tumors remains elusive. In this study, we demonstrate the structural basis for the binding of bromodomains (BDs) of bromodomain-containing protein 4 (Brd4) to AcLys310 and identify the BD inhibitor JQ1 as an effective small molecule to block this interaction. JQ1 suppresses TNF-α-mediated NF-κB activation and NF-κB-dependent target gene expression. In addition, JQ1 inhibits the proliferation and transformation potential of A549 lung cancer cells and suppresses the tumorigenicity of A549 cells in severe combined immunodeficiency mice. Furthermore, we demonstrate that depletion of Brd4 or treatment of cells with JQ1 induces the ubiquitination and degradation of the constitutively active nuclear form of RelA. Our results identify a novel function of Brd4 in maintaining the persistently active form of NF-κB found in tumors, and they suggest that interference with the interaction between acetylated RelA and Brd4 could be a potential therapeutic approach for the treatment of NF-κB-driven cancer.

The aim of this study was to establish and to optimize the preparation technology of whole blood internal quality control (IQC) products for blood transfusion compatibility testing. Several B-type RhD-negative blood samples collected from healthy donors were mixed. Two groups of whole blood IQC products, namely, the preservative solution group (PS group) and the saline group, were prepared. The agglutination intensity of IQC sample red cells and anti-B antibody, IgM anti-A antibody and reverse-typing A cell, IgG anti-D and O-type RhD-positive red cells, as well as free hemoglobin concentration in the supernatant of the two groups were detected. The erythrocytes in both groups were damaged to a certain extent during storage, but no evident (above moderate) hemolysis was observed in the stored sample within 42 days. The red cells remained structurally complete and the reaction activity of IgG anti-D reagent remained generally unchanged (P>0.05). Although the reaction activity oscillation of IgM anti-A reagent was observed, the agglutination intensity varied within an acceptable range of 1+. No difference was observed between the preparation methods of the samples, i.e., between the erythrocyte washed with saline and the one washed with red cell preservative solution (P>0.05). The long shelf life, low variance between tubes and stable antigen-antibody reaction activity of the whole blood IQC products prepared using the proposed method can meet the requirements of blood transfusion compatibility testing.

DNA repair helicases function in the cell to separate DNA duplexes or remodel nucleoprotein complexes. These functions are influenced by sensing and signaling; the cellular pool of a DNA helicase may contain subpopulations of enzymes carrying different post-translational modifications and performing distinct biochemical functions. Here, we report a novel experimental strategy, single-molecule sorting, which overcomes difficulties associated with comprehensive analysis of heterologously modified pool of proteins. This methodology was applied to visualize human DNA helicase F-box– containing DNA helicase (FBH1) acting on the DNA structures resembling a stalled or collapsed replication fork and its interactions with RAD51 nucleoprotein filament. Individual helicase molecules isolated from human cells with their native post-translational modifications were analyzed using total internal reflection fluorescence microscopy. Separation of the activity trajectories originated from ubiquitylated and non-ubiquitylated FBH1 molecules revealed that ubiquitylation affects FBH1 interaction with the RAD51 nucleoprotein filament, but not its translocase and helicase activities.

Helicobacter pylori infection causes chronic gastritis and peptic ulceration. H. pylori–initiated chronic gastritis is characterized by enhanced expression of many NF-κB–regulated inflammatory cytokines. Brd4 has emerged as an important NF-κB regulator and regulates the expression of many NF-κB–dependent inflammatory genes. In this study, we demonstrated that Brd4 was not only actively involved in H. pylori–induced inflammatory gene mRNA transcription but also H. pylori–induced inflammatory gene enhancer RNA (eRNA) synthesis. Suppression of H. pylori–induced eRNA synthesis impaired H. pylori–induced mRNA synthesis. Furthermore, H. pylori stimulated NF-κB–dependent recruitment of Brd4 to the promoters and enhancers of inflammatory genes to facilitate the RNA polymerase II–mediated eRNA and mRNA synthesis. Inhibition of Brd4 by JQ1 attenuated H. pylori–induced eRNA and mRNA synthesis for a subset of NF-κB–dependent inflammatory genes. JQ1 also inhibited H. pylori–induced interaction between Brd4 and RelA and the recruitment of Brd4 and RNA polymerase II to the promoters and enhancers of inflammatory genes. Finally, we demonstrated that JQ1 suppressed inflammatory gene expression, inflammation, and cell proliferation in H. pylori–infected mice. These studies highlight the importance of Brd4 in H. pylori–induced inflammatory gene expression and suggest that Brd4 could be a potential therapeutic target for the treatment of H. pylori–triggered inflammatory diseases and cancer.

One of the most fundamental and challenging questions in the cancer field is how immunity in patients with cancer is transformed from tumor immunosurveillance to tumor-promoting inflammation. Here, we identify the transcription factor STAT3 as the culprit responsible for this pathogenic event in lung cancer development. We found that antitumor type 1 CD4+ T-helper (Th1) cells and CD8+ T cells were directly counter balanced in lung cancer development with tumor-promoting myeloid-derived suppressor cells (MDSCs) and suppressive macrophages, and that activation of STAT3 in MDSCs and macrophages promoted tumorigenesis through pulmonary recruitment and increased resistance of suppressive cells to CD8+ T cells, enhancement of cytotoxicity toward CD4+ and CD8+ T cells, induction of regulatory T cell (Treg), inhibition of dendritic cells (DC), and polarization of macrophages toward the M2 phenotype. The deletion of myeloid STAT3 boosted antitumor immunity and suppressed lung tumorigenesis. These findings increase our understanding of immune programming in lung tumorigenesis and provide a mechanistic basis for developing STAT3-based immunotherapy against this and other solid tumors. Cancer Immunol Res; 5(3); 257–68. ©2017 AACR.

To analyze the data about red blood cell alloantibodies in patients from mainland China and to provide evidence for formulating a management guideline.The Chinese and English literatures about Chinese patients in mainland China published in periodicals were retrieved by CHKD, CNKI, CMJD and PubMed using the key words as unexpected antibody, irregular antibody, blood group antibody, hemolytic transfusion reaction (HTR), hemolytic disease of the newborn (HDN), hemolytic disease of the fetus and newborn (HDFN).A total of 5582 red blood cell alloantibodies were retrieved from 4800 patients. The average prevalence of alloantibody in 89 retrospective analysis reports was 0.34 %. Among all study patients, the 10 most common antibodies were anti-E (33.9%), anti-D (18.3%), anti-c (10.9%), anti-M (9.9%), anti-C (8.1%), anti-e (4.8%), anti-Le(a) (3.4%), anti-P1 (2.0%), anti-Mur (1.6%), and anti-Jk(a) (1.2%). Out of all 136 patients with HTR, the most frequentl alloantibodies were Rhesus antibodies (71.7%), and other antibodies included anti-Jk(b) (5.9%), anti-Le(a) (5.1%), anti-Jk(a) (3.7%), anti-M (1.5%), and anti-Mur (1.5%). A total of 644 alloantibodies contributing to HDFN come primarily from the Rhesus (93.1%) and MNS (6.0%) blood group systems.The postnatal Rh prophylaxis should become a routine procedure in mainland China. The use of blood matched for C, E, c, e, Jk(a) and Jk(b) should be recommended for Chinese patients with a history of multiple transfusions. Patients with MNS alloantibodies should be given sufficient attention, and Mur+ red blood cells should be included in antibody screening panels.

Transcription factor RUNX3 is inactivated in a number of malignancies, including breast cancer, and is suggested to function as a tumor suppressor. How RUNX3 functions as a tumor supressor in breast cancer remains undefined. Here, we show that about 20% of female Runx3+/− mice spontaneously developed ductal carcinoma at an average age of 14.5 months. Additionally, RUNX3 inhibits the estrogen-dependent proliferation and transformation potential of ERα-positive MCF-7 breast cancer cells in liquid culture and in soft agar and suppresses the tumorigenicity of MCF-7 cells in severe combined immunodeficiency (SCID) mice. Furthermore, RUNX3 inhibits ERα-dependent transactivation by reducing the stability of ERα. Consistent with its ability to regulate the levels of ERα, expression of RUNX3 inversely correlates with the expression of ERα in breast cancer cell lines, human breast cancer tissues and Runx3+/− mouse mammary tumors. By destabilizing ERα, RUNX3 acts as a novel tumor suppressor in breast cancer.

The eukaryotic transcription factor NF-kappaB regulates a wide range of host genes that control the inflammatory and immune responses, programmed cell death, cell proliferation and differentiation. The activation of NF-kappaB is tightly controlled both in the cytoplasm and in the nucleus. While the upstream cytoplasmic regulatory events for the activation of NF-kappaB are well studied, much less is known about the nuclear regulation of NF-kappaB. Emerging evidence suggests that NF-kappaB undergoes a variety of posttranslational modifications, and that these modifications play a key role in determining the duration and strength of NF-kappaB nuclear activity as well as its transcriptional output. Here we summarize the recent advances in our understanding of the posttranslational modifications of NF-kappaB, the interplay between the various modifications, and the physiological relevance of these modifications.

Posttranslational modifications of the RelA subunit of NF-kappaB, including acetylation and methylation, play a key role in controlling the strength and duration of its nuclear activity. Whether these modifications are functionally linked is largely unknown. Here, we show that the acetylation of lysine 310 of RelA impairs the Set9-mediated methylation of lysines 314 and 315, which is important for the ubiquitination and degradation of chromatin-associated RelA. Abolishing the acetylation of lysine 310 either by the deacetylase SIRT1 or by mutating lysine 310 to arginine enhances methylation. Conversely, enhancing the acetylation of lysine 310 by depleting SIRT1 or by replacing lysine 310 with acetyl-mimetic glutamine inhibits methylation, thereby decreasing ubiquitination, prolonging the stability of chromatin-associated RelA, and enhancing the transcriptional activity of NF-kappaB. The acetylation of lysine 310 of RelA interferes with its interaction with Set9. Based on structural modeling of the SET domain of Set9 with RelA, we propose that the positive charge of lysine 310 is critical for the binding of RelA to a negatively charged "exosite" within the SET domain of Set9. Together, these findings demonstrate for the first time an interplay between RelA acetylation and methylation and also provide a novel mechanism for the regulation of lysine methylation by acetylation.

Many of the pathologies linked to Helicobacter pylori are caused by the ability of the bacteria to induce chronic inflammation in the stomach of the host. One of the major transcription factors that regulate inflammation is NFκB, which is constitutively activated in many cancers including some gastric cancers. H. pylori has been shown to activate NFκB using several different bacterial components and host signaling pathways in cell-type and strain-specific ways. Our recent studies demonstrate that H. pylori utilizes its virulence factor CagA to target signaling molecule TAK1 for the activation of NFκB. In this article, we will summarize our findings together with other recent progress in the H. pylori-mediated activation of NFκB and discuss the role of CagA and TAK1 in the H. pylori-mediated activation of NFκB and gastric diseases.

Modification of eukaryotic proteins is a powerful strategy used by pathogenic bacteria to modulate host cells during infection. Previously, we demonstrated that Helicobacter pylori modify an unidentified protein within mammalian cell lysates in a manner consistent with the action of a bacterial ADP-ribosylating toxin. Here, we identified the modified eukaryotic factor as the abundant nuclear factor poly(ADP-ribose) polymerase-1 (PARP-1), which is important in the pathologies of several disease states typically associated with chronic H. pylori infection. However, rather than being ADP-ribosylated by an H. pylori toxin, the intrinsic poly(ADP-ribosyl) polymerase activity of PARP-1 is activated by a heat- and protease-sensitive H. pylori factor, resulting in automodification of PARP-1 with polymers of poly(ADP-ribose) (PAR). Moreover, during infection of gastric epithelial cells, H. pylori induce intracellular PAR-production by a PARP-1-dependent mechanism. Activation of PARP-1 by a pathogenic bacterium represents a previously unrecognized strategy for modulating host cell signaling during infection.

Helicobacter pylori-initiated chronic gastritis is characterized by the cag pathogenicity island-dependent upregulation of proinflammatory cytokines, which is largely mediated by the transcription factor nuclear factor (NF)-κB. However, the cag pathogenicity island-encoded proteins and cellular signalling molecules that are involved in H. pylori-induced NF-κB activation and inflammatory response remain unclear. Here, we show that H. pylori virulence factor CagA and host protein transforming growth factor-β-activated kinase 1 (TAK1) are essential for H. pylori-induced activation of NF-κB. CagA physically associates with TAK1 and enhances its activity and TAK1-induced NF-κB activation through the tumour necrosis factor receptor-associated factor 6-mediated, Lys 63-linked ubiquitination of TAK1. These findings show that polyubiquitination of TAK1 regulates the activation of NF-κB, which in turn is used by H. pylori CagA for the H. pylori-induced inflammatory response.