Negative regulation of TGF-ß by AMPK and implications in the treatment of associated disorders.

Transforming growth factor beta (TGF-ß) regulates a large number of biological processes, including proliferation, differentiation, immune response, and development. In addition, TGF-ß plays important roles in some pathological processes, for instance, it is upregulated and activated in fibrosis and advanced cancer. Adenosine monophosphate-activated protein kinase (AMPK) acts as a fuel gauge that is activated when cells sense shortage of ATP and increase in AMP or AMP:ATP ratio. Activation of AMPK slows down anabolic processes and stimulates catabolic processes, leading to increased production of ATP. Furthermore, the functions of AMPK have been extended beyond energy homeostasis. In fact, AMPK has been shown to exert a tumor suppressive effect. Recent studies have demonstrated negative impacts of AMPK on TGF-ß function. Therefore, in this review, we will discuss the differences in the biological functions of TGF-ß and AMPK, and some pathological processes such as fibrosis, epithelial-mesenchymal transition (EMT) and cancer metastasis, as well as angiogenesis and heterotopic ossifications where TGF-ß and AMPK exert opposite effects.

Expression profile and histological distribution of IFITM1 and IFITM3 during H9N2 avian influenza virus infection in BALB/c mice.

The H9N2 avian influenza virus is a pandemic threat which has repeatedly caused infection in humans and shows enhanced replication and transmission in mice. Previous reports showed that host factors, the interferon-inducible transmembrane (IFITM) protein, can block the replication of pathogens and affect their pathogenesis. BALB/c mice are routine laboratory animals used in influenza virus research, but the effects of H9N2 influenza virus on tissue distribution and expression pattern of IFITM in these mice are unknown. Here, we investigated the expression patterns and tissue distribution of IFITM1 and IFITM3 in BALB/c mice by infection with H9N2 AIV strains with only a PB2 residue 627 difference. The results showed that the expression patterns of ITITM1 and IFITM3 differ in various tissues of BALB/c mice at different time points after infection. IFITM1 and IFITM3 showed cell- and tissue-specific distribution in the lung, heart, liver, spleen, kidney and brain. Notably, the epithelial and neuronal cells all expressed the proteins of IFITM1 and IFITM3. Our results provide the first look at differences in IFITM1 and IFITM3 expression patterns in BALB/c mice infected by H9N2 influenza viruses. This will enhance research on the interaction between AIV and host and further will elucidate the pathogenesis of influenza virus infection based on the interferon-inducible transmembrane (IFITM) protein.

Expression pattern of NLRP3 and its related cytokines in the lung and brain of avian influenza virus H9N2 infected BALB/c mice.

BACKGROUND: H9N2 avian influenza virus (AIV) becomes the focus for its ability of transmission to mammals and as a donor to provide internal genes to form the new epidemic lethal influenza viruses. Residue 627 in PB2 has been proven the virulence factor of H9N2 avian influenza virus in mice, but the detailed data for inflammation difference between H9N2 virus strains with site 627 mutation is still unclear. The inflammasome NLRP3 is recently reported as the cellular machinery responsible for activation of inflammatory processes and plays an important role during the development of inflammation caused by influenza virus infection. METHODS: In this study, we investigated the expression pattern of NLRP3 and its related cytokines of IL-1ß and TNF-α in BALB/c mice infected by H9N2 AIV strains with only a site 627 difference at both mRNA and protein levels at different time points. RESULTS: The results showed that the expression level of NLRP3, IL-1ß and TNF-α changed in the lung and brain of BALB/c mice after infection by VK627 and rVK627E. The immunohistological results showed that the positive cells of NLRP3, IL-1ß and TNF-α altered the positive levels of original cells in tissues and infiltrated inflammatory cells which caused by H9N2 infection. CONCLUSIONS: Our results provided the basic data at differences in expression pattern of NLRP3 and its related cytokines in BALB/c mice infected by H9N2 influenza viruses with only a site 627 difference. This implied that NLRP3 inflammasome plays a role in host response to influenza virus infection and determines the outcome of clinical manifestation and pathological injury. This will explain the variable of pathological presentation in tissues and enhance research on inflammation process of the AIV H9N2 infection.

The correlation study between TOP2A gene expression in circulating tumor cells and chemotherapeutic drug resistance of patients with breast cancer.

BACKGROUND: Patients with breast cancer (BC) at advanced stages have poor outcomes because of high rate of recurrence and metastasis. Biomarkers for predicting prognosis remain to be explored. This study aimed to evaluate the relationships between circulating tumor cells (CTCs) and outcomes of BC patients. PATIENTS AND METHODS: A total of 50 female were enrolled in this study. Their diagnoses were determined by clinical characteristics, image data, and clinical pathology. CTC subtypes and TOP2A gene expression on CTCs were detected by CanPatrol™ technology and triple color in situ RNA hybridization (RNA-ISH), which divided into epithelial CTCs (eCTCs), mesenchymal CTCs (MCTCs), and hybrid CTCs (HCTCs) based on their surface markers. Hormone receptor, including estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER-2) expression, was measured by immunohistochemistry (IHC) method before treatment. The risk factors for predicting recurrence and metastasis were calculated by COX risk regression model. The progression-free survival (PFS) of patients was determined using Kaplan-Meier survival curve. RESULTS: The patients with a large tumor size (≥ 3 cm) and advanced tumor node metastasis (TNM) stages had high total CTCs (TCTCs) (P < 0.05). These patients also had high TOP2A expression level. COX risk regression analysis indicated that TOP2A expression levels in TCTCs, ER + , HER-2 + , and TNM stages were critical risk factors for recurrence and metastasis of patients (P < 0.05). The PFS of patients with ≥ 5 TCTCs, ≥ 3 HCTCs, and positive TOP2A expression in ≥ 3 TCTCs was significantly longer than that in patient with < 5 TCTCs, < 3 HCTCs, and TOP2A expression in < 3 TCTCs (P < 0.05). In contrast, the PFS of patients with positive hormone receptors (ER + , PR + , HER-2 +) also was dramatically lived longer than that in patients with negative hormone receptor expression. CONCLUSIONS: High TCTC, HCTCs, and positive TOP2A gene expression on CTCs were critical biomarkers for predicting outcomes of BC patients. Positive hormone receptor expression in BC patients has significant favor PFS.

Development of an Optoelectronic Integrated Sensor for a MEMS Mirror-Based Active Structured Light System.

Micro-electro-mechanical system (MEMS) scanning micromirrors are playing an increasingly important role in active structured light systems. However, the initial phase error of the structured light generated by a scanning micromirror seriously affects the accuracy of the corresponding system. This paper reports an optoelectronic integrated sensor with high irradiance responsivity and high linearity that can be used to correct the phase error of the micromirror. The optoelectronic integrated sensor consists of a large-area photodetector (PD) and a receiving circuit, including a post amplifier, an operational amplifier, a bandgap reference, and a reference current circuit. The optoelectronic sensor chip is fabricated in a 180 nm CMOS process. Experimental results show that with a 5 V power supply, the optoelectronic sensor has an irradiance responsivity of 100 mV/(µW/cm2) and a -3 dB bandwidth of 2 kHz. The minimal detectable light power is about 19.4 nW, which satisfies the requirements of many active structured light systems. Through testing, the application of the chip effectively reduces the phase error of the micromirror to 2.5%.

Evaluation and characterization of biochar on the biogeochemical behavior of polycyclic aromatic hydrocarbons in mangrove wetlands.

As the inter-tidal regions between land and ocean, mangrove ecosystems have high polycyclic aromatic hydrocarbons (PAHs) content, and the over accumulation of PAHs in mangrove wetland poses a serious ecological risk to the health of plant and living creatures. Comparison to the agricultural sources -biochar, biochar produced from wetland plant has lower O/C (molar ratio), larger N contents, higher stability and more benefits. However, whether the rhizosphere action occurs in biochar- amended sediment and how to influence the biogeochemical behavior of PAH have rarely been reported. In this context, a leaching procedure and pot experiment (60-d) were performed on migration and transformation of PAH at the sediment, and toxicity and their bioavailability in plant affected by the presence of Kandelia obovate-derived biochar in Southeast China. Root exudates amendments significantly increased the cumulative leaching-loss of pyrene by 36-51 % with or without biochar amendment via continuous diffusion and partition process, and biochar amendments decreased the bioavailability of pyrene (16.8-25.8 %) probably due to a faster pyrene sorption on inter-phase transport against desorption. The regression analysis indicated a significant relationship (p < 0.05) between leachate pH and pyrene concentrations. Notably, the bioaccumulation of pyrene on K. obovate parts had significant negative correlation (p < 0.05) to biochar. The activities of four key antioxidizes (phenylalanine ammonia-lyase, dismutases, peroxidases and catalases) were significantly decreased with the application of biochar. Moreover, biochar plays a positive role in cytochrome C release and phosphatidylserine secretion, and a combined biochar-rhizosphere approach can improve the stress tolerance and resistance of K. obovate with an enhanced synergetic effect, which could be a feasible remediation strategy for alleviating the mangrove sediment contaminated by PAH.

CEA cell adhesion molecule 5 enriches functional human hematopoietic stem cells capable of long-term multi-lineage engraftment.

Hematopoietic stem cell (HSC) surface markers improve the understanding of cell identity and function. Here, we report that human HSCs can be distinguished by their expression of the CEA Cell Adhesion Molecule 5 (CEACAM5, CD66e), which serves as a marker and a regulator of HSC function. CD66e+ cells exhibited a 5.5-fold enrichment for functional long term HSCs compared to CD66e- cells. CD66e+CD34+CD90+CD45RA- cells displayed robust multi-lineage repopulation and serial reconstitution ability in immunodeficient mice compared to CD66e-CD34+CD90+CD45RA-cells. CD66e expression also identified almost all repopulating HSCs within the CD34+CD90+CD45RA- population. Together, these results indicated that CEACAM5 is a marker that enriches functional human hematopoietic stem cells capable of long-term multi-lineage engraftment.

The Iron-Inflammation Axis in Early-Stage Triple-Negative Breast Cancer.

The iron-related homeostasis and inflammatory biomarker have been identified as prognostic factors for cancers. We aimed to explore the prognostic value of a novel comprehensive biomarker, the iron-monocyte-to-lymphocyte ratio (IronMLR) score, in patients with early-stage triple-negative breast cancer (TNBC) in this study. We retrospectively analysed a total of 257 early-stage TNBC patients treated at Sun Yat-sen University Cancer Center (SYSUCC) between March 2006 and October 2016. Their clinicopathological information and haematological data tested within 1 week of the diagnosis were collected. According to the IronMLR score cutoff value of 6.07 µmol/L determined by maximally selected rank statistics, patients were stratified into the low- and high-IronMLR groups, after a median follow-up of 92.3 months (95% confidence interval [CI] 76.0-119.3 months), significant differences in 5-years disease-free survival (DFS) rate (81.2%, 95% CI 76.2%-86.5% vs. 65.5%, 95% CI 50.3%-85.3%, p = 0.012) and 5-years overall survival (OS) rate (86.0%, 95% CI 81.6%-90.7% vs. 65.5%, 95% CI 50.3%-85.3%, p = 0.011) were seen between two groups. Further multivariate Cox regression analysis revealed the IronMLR score as an independent predictor for DFS and OS, respectively, we then established a prognostic nomogram integrating the IronMLR score, T stage and N stage for individualized survival predictions. The prognostic model showed good predictive performance with a C-index of DFS 0.725 (95% CI 0.662-0.788) and OS 0.758 (95% CI 0.689-0.826), respectively. Besides, calibration curves for 1-, 3-, 5-DFS, and OS represented satisfactory consistency between actual and nomogram predicted survival. In conclusion, the Iron-inflammation axis might be a potential prognostic biomarker of survival outcomes for patients with early-stage TNBC, prognostic nomograms based on it with good predictive performance might improve individualized survival predictions.

Metformin Inhibits Chemokine Expression Through the AMPK/NF-κB Signaling Pathway.

Inflammation is mediated by cytokines and chemokines, which are considered targets of inflammatory diseases. Mounting evidence has demonstrated the anti-inflammatory benefits of metformin. However, the underlying mechanisms are not completely understood. In this study, we aim to elucidate the regulatory effects of metformin on chemokine expression and the possible mechanisms using RAW264.7 cells, a mouse macrophage cell line, as a model. First, we treated the cells with lipopolysaccharide (LPS), and found that the expression of CXCL10 and CXCL11 was markedly induced in a dose- and time-dependent fashion concurrent with the inhibition of AMPK activity. Then, we treated the cells with metformin, and analyzed the expression of CCL2, CXCL10, and CXCL11 by quantitative real-time polymerase chain reaction (PCR). We observed that metformin prevented the stimulating effect of LPS on these chemokines as well as IL-1 and IL-6. Second, the inhibitory effects of metformin on LPS-induced chemokine expression were diminished by Compound C, a chemical inhibitor of AMPK. Finally, we investigated whether the NF-κB signaling pathway is regulated by metformin in this setting. Our results showed that metformin inhibited the phosphorylation of I-κBα and p65 while it activated AMPK. Therefore, the results suggest that metformin inhibits LPS-induced chemokine expression through the AMPK and NF-κB signaling pathways.

Tissue-specific expression pattern and histological distribution of NLRP3 in Chinese yellow chicken.

Pathogen-associated molecular patterns (PAMPs) play important role in inflammation which means response of the host to stimuli. NOD-like receptor protein 3 (NLRP3) inflammasome is involved in the onset and development of inflammation. NLRP3, as one of the most important inflammasome sensors, has significant effect on the regulation of inflammasome activation to avoid the consequences of over activation. Up to date, there are no detailed tissue specific expression and distribution data about NLPR3 in chicken. Here, NLRP3 of Chinese yellow chicken was cloned and sequence analyzed, the polyclonal antibody was produced by purified protein of recombinant prokaryotic expression. Relative expression levels and tissue distribution of NLRP3 were investigated by real-time quantitative PCR and immunohistochemical analysis, respectively. The results showed that NLRP3 gene is highly variable between mammalian and avian. The nucleotide homology of NLRP3 between yellow chicken and Bos taurus, Hainan black goat, Sus scrofa, Callithrix jacchus, Homo sapiens, Macaca mulatta, Mus musculus and Rattus norvegicus were 54.2%, 53.9%, 53.7%, 55.4%, 54.3%, 54.5%, 53.5% and 53.7%. NLRP3 expressed in all detected tissues and higher in the trachea are lung than in other tissues. Cytoplasmic expression of NLRP3 was detected in ciliated epithelial cells, basal cells and cells in lamina propria of trachea, alveolar epithelial cells, cardiac muscle cells, cerebral cortex neurons, epithelial reticular cells of the spleen, and lymphocytes of medulla in stannius follicle, liver cells and the renal tubule epithelial cells. The results will help to elucidate the role of NLRP3 of different tissues in inflammatory diseases of chicken and provide a basis for further investigations in the function and evolution of NLRP3 in different species, which would be helpful for further research on avian inflammatory diseases.