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Purpose: Diffuse large B-cell lymphoma (DLBCL) is the most common pathological type of non-Hodgkin lymphoma (NHL) and is closely associated with hepatitis B virus (HBV) infection and hepatitis B X (HBx) gene integration. This project investigates the cellular biological effects and molecular mechanisms responsible for lymphomagenesis and the progression of HBx gene integration in DLBCL. Methods: The integration of the HBx gene was monitored using sequencing technology, and variations in cell biological effects and related molecular mechanisms were investigated using exvivocell experiments, which were verified in experimental animals and clinical cases. Results: The data shows that clinical DLBCL cells demonstrate HBx integration, and the sequencing analysis validated enabled successfully constructed HBx transfected cells. Compared with control cells, HBx transfected cells had a significantly reduced proportion of mitochondrial membrane potential, obviously reduced signals of chromosome DNA breaks, and proportion of apoptotic cells. Therefore, HBx integration was able to cause the biological outcome of apoptosis reduction. Further studies found that this decreased apoptosis level is associated with a significantly reduced downstream of cleaved Caspase-3 and poly ADP-ribose polymerase (PARP) proteins, revealing the molecular mechanisms of HBx-associated apoptosis in DLBCL. The animal experiments also demonstrated that the protein expression of cleaved Caspase-3 and PARP was prominently reduced in HBx transfected cells from a subcutaneous tumor in nude mice. Overall, the clinically observed expression of cleaved PARP proteins in tissues of HBx-positive DLBCL patients is lower than in HBx-negative patients. Conclusion: In DLBCL, HBx gene integration inhibits cell apoptosis through the Caspase-3-PARP pathway.

Background In recent years, the application of immunotherapy combined with chemotherapy in the first-line lung cancer has showed significant benefit in improving long-term survival. Immunotherapy also has risks of immune-related pneumonitis (IRP) after long-term treatment. Despite the treatment strategy of the IRP has been very clear. However, the mechanism is unclear. Case presentation A 73-year-old male patient was diagnosed with left lung adenocarcinoma IVa, EGFR, ALK, ROS1 negative. The patient received anti-PD1 antibody combined with pemetrexed and cisplatin. After 5 cycles of treatment, partial response was obtained. Subsequently, the patient continued the treatment of anti-PD1 antibody combined with pemetrexed. Before the 7th cycle, the CT found a new lesion in the basal segment of the right lower lobe. It was diagnosed with IRP and pneumocystis jirovecii. The patient did not give trimethoprim–sulphamethoxazole (TMP–SMX) and corticosteroids, symptoms and radiological lesions had improved. We describe the report of immune-related pneumonitis trigged by anti PD-1 and monitored the dynamic changes of CD4+, CD8+ T lymphocytes, MDSC and Treg cells in the bilateral bronchoalveolar alveolar lavage fluid. From the point of view of immune cells, the mechanism of immune reconstitution inflammatory syndrome is confirmed. Based on the current case report and literature, this study proposes a potential mechanism of the onset. Conclusion Immune reconstitution inflammatory syndrome may be potential mechanism of IRP. This study may improve our understanding of the pathogenesis underlying IRP. We believe the detection and dynamic monitoring CD4+, CD8+ T lymphocytes, MDSC and Treg cells can provide more accurate procedures.

Inflammatory cytokine‐induced cell apoptosis is important for initiation and progression of chronic heart failure (CHF). Non‐coding RNAs, including long non‐coding RNAs and microRNAs, have emerged as critical regulators of this pathological process. The role in regulating inflammation and induction to cell apoptosis in CHF is not well understood. This study found CHF patients had elevated serum miR‐939‐5p, with greater increase in New York Heart Association (NYHA) I‐II patients than in NYHA III‐IV. Moreover, miR‐939‐5p was positively correlated with B‐type natriuretic peptide (BNP) in NYHA III‐IV patients, while not in NYHA I‐II. Further study showed miR‐939‐5p mimics promoted cell proliferation and inhibited inflammatory cytokine‐induced apoptosis of HUVECs and H9C2, while inhibition of endogenous miR‐939‐5p produced the opposite effects. Induced nitric oxide synthase (iNOS) and tumour necrosis factor α (TNFα) were identified as target genes of miR‐939‐5p. Additionally, lncRNA‐NOS2P3 acted as an endogenous sponge RNA to inhibit miR‐939‐5p expression, regulate the expression of iNOS/TNFα and control inflammation‐induced cells apoptosis. These suggest that CHF patients exhibited elevated serum miR‐939‐5p level especially in NYHA I‐II grades. And lnc‐NOS2P3‐miR‐939‐5p‐iNOS/TNFα pathway regulated inflammatory cytokine‐induced endothelial and myocardial cells apoptosis and provided a promising strategy for diagnosis and treatment of CHF.

Purpose This study aims to elucidate the biological behavior of Neuritin abnormal expression in pulmonary vascular endothelial cells (VECs) of non-small cell lung cancer (NSCLC), and explore its possible underlying mechanisms. Patients and methods Primary NSCLC-VECs were isolated from 10 cancer tissues from NSCLC patients, purified and identified by CD34 and Factor VIII staining. Real-time PCR and Western-blot were adopted for detecting the expression levels of Neuritin, Notch1, and VEGFR in NSCLC-VECs and HPMECs. Neuritin-overexpression, Neuritin-knockdown NSCLC-VECs and HPMECs were constructed by transfection of pcDNA3, 1-Neuritin vector, and pBS/U6-Neuritin siRNA. Changes in cell proliferation, migration, cell cycle, and apoptosis were determined by using the MTT assay, scratch assay, transwell migration assay, and flow cytometry, respectively. Post-transfection changes in cell morphology were examined by scanning electron microscopy. Results The expression of Neuritin in NSCLC-VECs was significantly higher compared to that in HPMECs (p

Background: Hypoxia, a common feature of rheumatoid arthritis (RA), induces the overexpression of peptidyl arginine deiminase 4 (PADI4) in fibroblast-like synoviocytes (FLSs) and macrophages. However, the roles of PADI4 and its inducer hypoxia in the regulation of macrophage polarization remain unclear. This study aimed to investigate the role of hypoxia-PADI4 for macrophage polarization in RA patients.Methods: synovial tissue(ST) and synovial fluid(SF) were collected from 3 OA patients and 6 RA patients. The distribution of M1 and M2 in ST and cytokines in SF were examined by immunohistochemical analysis and BioPlex immunoassays. THP-1 macrophages and BMDM polarization were determined under normoxic (21% oxygen) or hypoxic (3% oxygen) conditions. The effects of PADI4 on macrophages were determined by transfection of adenovirus vector-coated PADI4 (AdPADI4) and the use of PADI4 inhibitor. To mimic the environment of RA joints, THP-1 macrophages polarization was examined after coculturing with RA-FLSs. Finally, the roles of PADI4 in joint synovial lesions on macrophage polarization were investigated in collagen-induced arthritis(CIA) rats.Results: We found increased macrophage polarization of M1 and M2 in the RA ST, compared with OA ST. The ratio of M1/M2 for RA and OA was 1.633 ± 0.1443 and 2.544 ± 0.4429, respectively. The concentration of M1- and M2-type cytokines was higher in RA than that in OA patients. Hypoxia contributed to the increase of the gene and protein expression of M1 and M2 markers. M1- but not M2-type gene expression showed a positive relationship with PADI4 expression while the level of expression of M2-type genes showed no significant difference. RA-FLSs could promote the copolarization of M1 and M2, and PADI4 inhibitor reversed M1 activation. The degree of joint swelling and destruction was effectively alleviated, and the number of macrophages especially M1 decreased in CIA rats after downregulating PADI4 expression.Conclusion: Hypoxia is responsible for the copolarization of M1 and M2. Hypoxia-associated PADI4 is responsible for M1 macrophage activation, implying that inflammatory environment can be eased by decreasing PADI4 expression and improving the hypoxic environment.

Room-temperature superconductivity has been attracting increasing attention in recent years. Recent studies have proved the potential of compressed H-rich materials for achieving room-temperature superconductivity. In this paper, we study the phononic thermal transport in the rare earth yttrium hydrides allotropes under 0 GPa, 50 GPa, and 300 GPa by using Boltzmann transport equation. We find that the lattice thermal conductivity of yttrium hydrides increases with the pressure among different allotropes, which is attributed to the increase of bond strength and the decrease of phonon-phonon scattering due to structural compression. Yttrium hydrides structure at high pressure of 300 GPa is the superconducting phase, and has high thermal conductivity around 1360 Wm-1K-1 at room temperature. Comparison of phonon properties with existing high thermal conductivity materials further uncovers the origin for the observed high thermal conductivity. For the zero pressure allotrope, a large number of optical flat bands mix with the low-frequency acoustic phonons, which significantly increases the phonon scattering channel and effectively suppresses the phonon lifetime. As for yttrium hydrides allotropes under 50 GPa and 300 GPa, there are two obvious band gaps in the phonon dispersion relation, and the band gap of the structure at 300 GPa is significantly wider. The occurrence of the band gap effectively inhibits the absorption and emission process of the three-phonon interactions, leading to the decrease of phonon scattering and thus the increase of the phonon lifetime and thermal conductivity at high pressure. Our work reveals the physical mechanism of the thermal transport behaviors in yttrium hydrides structures under different pressures.

Background: Inflammatory cytokines induced cells apoptosis is important for initiation and progression of chronic heart failure (CHF). Long noncoding RNAs (lncRNAs) and microRNAs (miRNAs) were critical in this pathogenesis. However, their roles in inflammation and apoptosis of CHF remain unclear. Methods: A total of 75 CHF and 36 non-CHF control patients were collected and the level of miR-939-5p was detected. Bioinformatics analysis and luciferase reporter assay was used to predict and verify pairs of lncRNA-miRNA-mRNA. RT-qPCR was used to evaluate the expression of lncRNA, miRNAs and mRNAs. CCK8 and flow cytometry were applied to determine cells vitality and apoptosis. Western blot was used for protein level analysis. Results : CHF patients had elevated serum miR-939-5p, with greater increase in NYHA I-II patients than in NYHA III-IV. Moreover, miR-939-5p was positively correlated with BNP in NYHA III-IV patients, while not in NYHA I-II, suggesting that miR-939-5p might be an additional supplement for diagnosis of heart failure. Further study showed miR-939-5p mimic promoted proliferation and inhibited inflammatory cytokines induced apoptosis of HUVECs and H9C2, while inhibition of endogenous miR-939-5p by antagomir produced the opposite effects. INOS and TNFα were identified and confirmed as target genes of miR-939-5p. Moreover, we identified lncRNA-NOS2P3 as a sponge RNA to inhibit miR-939-5p expression, regulate the expression of iNOS/TNFα, and control inflammation induced myocardial and endothelial cells apoptosis. Conclusions : Together, CHF patients exhibited elevated serum miR-939-5p especially in NYHA I-II grades. Lnc-NOS2P3-miR-939-5p-iNOS/TNFα pathway played an important role in regulating inflammatory cytokines induced cells apoptosis and provided a promising strategy for diagnosis and treatment of CHF.

Qiao Zhang,1 Juan Zhang,2 Jian Zhang,3 Patiguli Aerxiding,1 Amina Quhai,1 Cuncun Chen,4 Li Shan1 1Department of Thoraciconcology, The Third Affiliated Hospital of Xinjiang Medical University, Tumor Hospital Affiliated to Xinjiang Medical University, Urumqi, Xinjiang 830011, People’s Republic of China; 2Department of Respiratory and Critical Care Medicine, First People’s Hospital of Kashgar, Xinjiang 844000, People’s Republic of China; 3Health Corps of the People’s Liberation Army 69260 Troops, Urumqi, Xinjiang, 830002, People’s Republic of China; 4Department of Thoracic Surgery, Chest Hospital of Xinjiang Uyghur Autonomous Region, Urumqi, Xinjiang 830049, People’s Republic of ChinaCorrespondence: Cuncun ChenDepartment of Thoracic Surgery, Chest Hospital of Xinjiang Uyghur Autonomous Region, Urumqi, Xinjiang 830049, People’s Republic of ChinaTel +0991-7500419Fax +860991-7860085Email chenpuyingxiong@yeah.net Li ShanDepartment of Thoraciconcology, The Third Affiliated Hospital of Xinjiang Medical University, Tumor Hospital Affiliated to Xinjiang Medical University, Urumqi, Xinjiang 830011, People’s Republic of ChinaTel +8613609989394Fax +860991-7819342Email shanlinew319@163.comPurpose: This study aims to elucidate the biological behavior of Neuritin abnormal expression in pulmonary vascular endothelial cells (VECs) of non-small cell lung cancer (NSCLC), and explore its possible underlying mechanisms.Patients and methods: Primary NSCLC-VECs were isolated from 10 cancer tissues from NSCLC patients, purified and identified by CD34 and Factor VIII staining. Real-time PCR and Western-blot were adopted for detecting the expression levels of Neuritin, Notch1, and VEGFR in NSCLC-VECs and HPMECs. Neuritin-overexpression, Neuritin-knockdown NSCLC-VECs and HPMECs were constructed by transfection of pcDNA3, 1-Neuritin vector, and pBS/U6-Neuritin siRNA. Changes in cell proliferation, migration, cell cycle, and apoptosis were determined by using the MTT assay, scratch assay, transwell migration assay, and flow cytometry, respectively. Post-transfection changes in cell morphology were examined by scanning electron microscopy.Results: The expression of Neuritin in NSCLC-VECs was significantly higher compared to that in HPMECs (p

Background: Inflammatory cytokines induced cells apoptosis is important for initiation and progression of chronic heart failure (CHF). Long noncoding RNAs (lncRNAs) and microRNAs (miRNAs) were critical in this pathogenesis. However, their roles in inflammation and apoptosis of CHF remain unclear. Methods: Bioinformatics analysis and luciferase reporter assay was used to predict and verify pairs of lncRNA-miRNA-mRNA. RT-qPCR was used to evaluate the expression of lncRNA, miRNAs and mRNAs. CCK8 and flow cytometry were applied to determine cells vitality and apoptosis. Western blot was used for protein level analysis. Findings: CHF patients had elevated serum miR-939-5p, with greater increase in NYHA I-II patients than in NYHA III-IV. Moreover, miR-939-5p was positively correlated with BNP in NYHA III-IV patients, while not in NYHA I-II, suggesting that miR-939-5p might be an additional supplement for diagnosis of heart failure. Further study showed miR-939-5p mimic promoted proliferation and inhibited inflammatory cytokines induced apoptosis of HUVECs and H9C2, while inhibition of endogenous miR-939-5p by antagomir produced the opposite effects. INOS and TNFα were identified and confirmed as target genes of miR-939-5p. Moreover, we identified lncRNA-NOS2P3 as a sponge RNA to inhibit miR-939-5p expression, regulate the expression of iNOS/TNFα, and control inflammation induced myocardial and endothelial cells apoptosis. Interpretation: Together, CHF patients exhibited elevated serum miR-939-5p especially in NYHA I-II grades. Lnc-NOS2P3-miR-939-5p-iNOS/TNFα pathway played an important role in regulating inflammatory cytokines induced cells apoptosis and provided a promising strategy for diagnosis and treatment of CHF. Funding: National Natural Science Foundation of China, Foundation of Shanghai Municipal Health Commission, Project of Tongji University. Declaration of Interest: The authors declared that they have no conflict of interest. Ethical Approval: The Institutional Review Board of Tongji Medical School affiliated Shanghai East Hospital approved the study protocol and the written informed consent was obtained from each participant before any sample or data collection.

Background Lipoprotein (a) [Lp(a)], which is genetically determined by the LPA gene kringle IV type 2 (KIV-2) repeat copy number, has previously been reported in different populations. However, it is uncertain if the same occurs in the Chinese Han population. This study explored the correlation of Lp(a) mass or particle concentration with KIV-2 repeat copy number and application for coronary atherosclerotic heart disease (CAHD) risk assessment. Methods A cross-sectional study including 884 subjects was conducted. The Lp(a) level and routine risk factors of CAHD were compared. The KIV-2 copy number distribution, relationship with Lp(a), and assessment for CAHD risk were explored. Results The mean of Lp(a) mass or particle concentration in the CAHD group was higher than that in the non-CAHD group, while the KIV-2 copy number in the CAHD group was lower. Lp(a) had auxiliary values in gauging the type of plaque and was significantly higher in the soft-plaque group than that in the other two groups (200 mg/L [21.5 nmol/L], 166 mg/L [18.6 nmol/L], 149 mg/L [17.1 nmol/L], respectively, P

Multi-layered mechanisms of virus host interaction exist for chronic hepatitis B virus (HBV) infection, which have been typically manifested at the microRNA level. Our previous study suggested that miRNA-939 (miR-939) may play a potential role in regulating HBV replication. Here we further investigated the mechanism by which miR-939 regulates HBV life cycle. We found that miR-939 inhibited the abundance of viral RNAs without direct miRNA-mRNA base pairing, but via host factors. Expression profiling and functional validation identified Jmjd3 as a target responsible for miR-939 induced anti-HBV effect. Jmjd3 appeared to enhance the transcription efficiency of HBV enhancer II/core promoter (En II) in a C/EBPα-dependent manner. However, the demethylase activity of Jmjd3 was not required in this process. Rather, Jmjd3’s transactivation activity depended on its interaction with C/EBPα. This coordinated action further recruited the Brm containing SWI/SNF chromatin remodeling complex which promoted the transcription of HBV RNAs. Taken together, we propose that the miR-939-Jmjd3 axis perturbs the accessibility of En II promoter to essential nuclear factors (C/EBPα and SWI/SNF complex) therefore leading to compromised viral RNA synthesis and hence restricted viral multiplication.

Background Interferon (IFN) and pegylated interferon (PEG-IFN) treatment of chronic hepatitis B leads to a sustained virological response in a limited proportion of patients and has considerable side effects. To find novel markers associated with prognosis of IFN therapy, we investigated whether a pretreatment plasma microRNA profile could be used to predict early virological response to IFN. Methods We performed microRNA microarray analysis of plasma samples from 94 patients with chronic hepatitis B who received IFN therapy. The microRNA profiles from 13 liver biopsy samples were also measured. The OneR feature ranking and incremental feature selection method were used to rank and optimize the number of features in the model. Support vector machine prediction engine and jack-knife cross-validation were used to generate and evaluate the prediction model. Results The optimized model consisting of 11 microRNAs yielded a 74.2% overall accuracy in the training group and was independently confirmed in the test group (71.4% accuracy). Univariate and multivariate logistic regression analyses confirmed its independent association with early virological response (OR=7.35; P=2.12x10-5). Combining the microRNA profile with the alanine aminotransferase level improved the overall accuracy from 73.4% to 77.3%. Co-transfection of an HBV replicative construct with microRNA mimics revealed that let-7f, miR-939 and miR-638 were functionally associated with the HBV life cycle. Conclusions The 11 microRNA signatures in plasma, together with basic clinical variables, might provide an accurate method to assist in medication decisions and improve the overall sustained response to IFN treatment.

The hepatitis B virus (HBV) is a DNA virus that can cause chronic hepatitis B (CHB) in humans. Current therapies for CHB infection are limited in efficacy and do not target the pre-existing viral genomic DNA, which are present in the nucleus as a covalently closed circular DNA (cccDNA) form. The transcription activator-like (TAL) effector nucleases (TALENs) are newly developed enzymes that can cleave sequence-specific DNA targets. Here, TALENs targeting the conserved regions of the viral genomic DNA among different HBV genotypes were constructed. The expression of TALENs in Huh7 cells transfected with monomeric linear full-length HBV DNA significantly reduced the viral production of HBeAg, HBsAg, HBcAg, and pgRNA, resulted in a decreased cccDNA level and misrepaired cccDNAs without apparent cytotoxic effects. The anti-HBV effect of TALENs was further demonstrated in a hydrodynamic injection-based mouse model. In addition, an enhanced antiviral effect with combinations of TALENs and interferon-α (IFN-α) treatment was observed and expression of TALENs restored HBV suppressed IFN-stimulated response element-directed transcription. Taken together, these data indicate that TALENs can specifically target and successfully inactivate the HBV genome and are potently synergistic with IFN-α, thus providing a potential therapeutic strategy for treating CHB infection.

Treatment with exogenous interferon (IFN)-α is not effective in the majority of patients with chronic hepatitis B virus (HBV) infection. Recent evidence suggests that HBV has evolved strategies to block the nuclear translocation of signal transducer and activator of transcription (STAT) 1 to limit IFN-α–induced cellular antiviral responses. However, it remains unclear whether STAT1 translocation is impaired in chronic hepatitis B patients and what mechanisms are involved. Here we report that the expression of HBV polymerase (Pol) in human hepatic cell lines inhibited induction of IFN-stimulated genes and resulted in a weakened antiviral activity of IFN-α. Ectopic expression of Pol suppressed IFN-α–induced STAT1 serine 727 phosphorylation and STAT1/2 nuclear accumulation, whereas STAT1 tyrosine 701 phosphorylation, and STAT1-STAT2 heterodimer formation were not affected. Further studies demonstrated that Pol interacted with the catalytic domain of protein kinase C-δ (PKC-δ) and perturbed PKC-δ phosphorylation and its association with STAT1, which resulted in the suppression of STAT1 Ser727 phosphorylation. Moreover, Pol was found to interfere with nuclear transportation of STAT1/2 by competitively binding to the region of importin-α5 required for STAT1/2 recruitment. Truncation analysis suggested that the terminal protein and RNase H domains of Pol were able to bind to PKC-δ and importin-α5, respectively, and were responsible for the inhibition of IFN-α signaling. More importantly, the inhibition of STAT1 and PKC-δ phosphorylation were confirmed in a hydrodynamic-based HBV mouse model, and the blockage of IFN-α–induced STAT1/2 nuclear translocation was observed in HBV-infected cells from liver biopsies of chronic HBV patients. Conclusions: These results demonstrate a role for Pol in HBV-mediated antagonization of IFN-α signaling and provide a possible molecular mechanism by which HBV resists the IFN therapy and maintains its persistence. (HEPATOLOGY 2013;)