Insights into autotrophic carbon fixation strategies through metagonomics in the sediments of seagrass beds.

Seagrass beds contributes up to 10% ocean carbon storage. Carbon fixation in seagrass bed greatly affect global carbon cycle. Currently, six carbon fixation pathways are widely studied: Calvin, reductive tricarboxylic acid (rTCA), Wood-Ljungdahl (WL), 3-hydroxypropionate (3HP), 3-hydroxypropionate/4-hydroxybutyrate (3HP/4HB) and dicarboxylate/4-hydroxybutyrate (DC/4-HB). Despite the knowledges about carbon fixation increase, the carbon fixation strategies in seagrass bed sediment remain unexplored. We collected seagrass bed sediment samples from three sites with different characteristics in Weihai, a city in Shandong, China. The carbon fixation strategies were investigated through metagenomics. The results exhibited that five pathways were present, of which Calvin and WL were the most dominant. The community structure of microorganisms containing the key genes of these pathways were further analyzed, and those dominant microorganisms with carbon fixing potential were revealed. Phosphorus significantly negatively corelated with those microorganisms. This study provides an insight into the strategies of carbon fixation in seagrass bed sediments.

CGB5, INHBA and TRAJ19 Hold Prognostic Potential as Immune Genes for Patients with Gastric Cancer.

BACKGROUND: Gastric cancer (GC) seriously threatens people's health and life quality worldwide. AIM: The current study sought to explore prognostic immune genes and their regulatory network in GC. METHODS: First, expression data in GC and normal samples were analyzed based on bioinformatics analysis. Immune-related genes were identified and confirmed with univariate/multivariate Cox analysis and receiver-operating characteristic curve. The upstream transcription factors of immune genes were subsequently predicted, and their regulatory network was constructed. GC and adjacent normal tissues were obtained from 76 patients with GC to determine the expression patterns of immune genes and their correlation with overall prognosis. CD8+ T-cell infiltration of patients with high or low risk was detected by means of immunohistochemistry. RESULTS: Bioinformatics analysis highlighted 3689 differentially expressed genes in GC, including 87 immune genes, 8 of which were significantly associated with patient survival. CGB5 and INHBA were high-risk genes, while TRAJ19 was identified as a low-risk gene, all of which were found to be regulated by 11 different transcription factors. Furthermore, CGB5 and INHBA exhibited negative correlation with the prognosis of GC patients; however, TRAJ19 was positively correlated with GC patient prognosis. The incidence of lymph node metastasis was higher, the pathological stage was advanced and the infiltrated CD8+ T cells were fewer in the high-risk GC group. CONCLUSIONS: Overall, our findings identified the key roles of CGB5, INHBA and TRAJ19 in prognosis GC patients, serving as an important gene set for prognostic prediction.

Jujuboside A ameliorates tubulointerstitial fibrosis in diabetic mice through down-regulating the YY1/TGF-ß1 signaling pathway.

Diabetic nephropathy (DN) is one of the most common complications of diabetes mellitus, which is characterized in renal tubulointerstitial fibrosis (TIF). The current study was designed to investigate the protective effect of Jujuboside A (Ju A) on TIF in type 2 diabetes (T2DM) mice, and explore its underlying anti-fibrosis mechanism. A mouse T2DM model was established using high fat diet (HFD) feeding combined with intraperitoneal injection of streptozotocin (STZ). Then, diabetic mice were treated with Ju A (10, 20 and 40 mg·kg-1·d-1, i.g.) for 12 weeks. Results showed that administration of Ju A not only down-regulated fasting blood glucose (FBG) levels, but also improved hyperlipidemia and renal function in diabetic mice. Moreover, the reduced ECM accumulation was observed in the renal cortex of Ju A treated diabetic mice, while the TIF progression was also attenuated by Ju A through blocking the epithelial-to-mesenchymal transition (EMT) of renal tubular epithelial cells (RTECs). Further mechanism studies showed that Ju A treatment effectively down-regulated the protein expression and subsequent nuclear translocation of Yin Yang 1 (YY1) in the renal cortex of diabetic mice, and reduced the levels of transforming growth factor-ß1 (TGF-ß1) in the serum and renal cortex of Ju A treated mice. According to invitro studies, the up-regulated YY1/TGF-ß1 signaling pathway was restored by Ju A in high glucose (HG) cultured HK-2 cells. Taken together, these findings demonstrated that Ju A can ameliorate the TIF of DN through down-regulating the YY1/TGF-ß1 signaling pathway.

KDM4B promotes gastric cancer metastasis by regulating miR-125b-mediated activation of Wnt signaling.

Emerging evidence has demonstrated that the aberrant expression of histone-modifying enzymes such as histone demethylases contributes to gastric carcinogenesis and progression. The role of KDM4B in cancer progression has been gradually revealed. However, the underlying mechanisms regulating gastric cancer metastasis of KDM4B remain unclear. In the present study we determined KDM4B expression in gastric cancer and its biologic function in vitro and in vivo. We found that KDM4B expression was significantly increased in most gastric cancer tissues compared with the adjacent normal tissues. Upregulated expression of KDM4B in human gastric cancer was correlated with poor prognosis. In vitro, KDM4B overexpression in AGS cells promoted cell invasion, whereas knockdown of KDM4B inhibited cell invasion. Furthermore, KDM4B overexpression also promoted tumor metastasis in vivo. Mechanistically, KDM4B upregulated miR-125b expression and activated Wnt signaling pathway. More important, miR-125b partially mediated KDM4B-induced activation of Wnt signaling. Finally, we demonstrated that KDM4B promoted gastric cancer cell invasion in vitro and cancer metastasis in vivo, at least in part, by upregulating miR-125b expression. These data provided novel insights on the role of KDM4B-driven gastric cancer metastasis and indicated that KDM4B may be served as a potential target for gastric cancer.

Meta-Analysis for the Therapeutic Effect of Neoadjuvant Therapy in Resectable Esophageal Cancer.

We aimed to review the therapeutic effects of neoadjuvant chemoradiotherapy (NCRT), chemotherapy (NCT), and radiotherapy (NRT) on patients with resectable Esophageal cancer (EsC) by comparison with surgery alone (SA). PubMed, EMBASE and Cochrane were searched for eligible studies published up to March 2015. Cochrane reviews were used for quality assessment. Eight primary outcomes were analyzed. Risk ratios (RRs)/ hazard ratios (HRs) and corresponding 95% confidence intervals (95% CIs) were calculated using the random- or fixed- effects model. Heterogeneity was assessed using the Chi-square-based Q statistic and the I 2 test. Publication bias was examined by the Begg's funnel plot. Totally 24 articles including 4718 EsC cases were eligible for this meta-analysis. The quality of the literatures was relatively high. Significant difference was found in five-year survival rate (RR = 1.45, 95% CI: 1.17-1.79, P < 0.01) between patients treated with NCT and SA, while the eight enrolled primary outcomes were all statistically different between NCRT and SA, and significant difference was identified in three-year survival between NCRT and NCT (RR = 1.35, 95% CI: 1.14-1.60, P < 0.01). No obvious publication bias was observed. NCRT and NCT provide an obvious benefit for EsC treatment over SA, and NCRT possesses a clear advantage compared with NCT.

Na+/H+ exchanger 1, Na+/Ca2+ exchanger 1 and calmodulin complex regulates interleukin 6-mediated cellular behavior of human hepatocellular carcinoma.

Interleukin 6 (IL6) is a key cytokine involved in the development and progression of inflammation-associated hepatocellular carcinoma (HCC). However, the mechanisms of IL6 action on HCC remain largely unknown. Proton and Ca(2+) are two intracellular messenger ions, which are believed to play a central role in tumorigenesis and tumor progression. In this study, we found that IL6 stimulation markedly increased intracellualr pH recovery rates of human HCC cells, Huh7 and HepG2, after NH4Cl acidification, and the NH4Cl acidification induced transient intracellular Ca(2+) increases in the HCC cells. The inhibition of Na(+)/H(+) exchanger 1 (NHE1), Na(+)/Ca(2+) exchanger 1 (NCX1) and calmodulin (CaM) inhibited the IL6 stimulation-induced intracellular pH recovery increases and NH4Cl acidification-induced intracellular Ca(2+) increases. IL6 stimulation also induced the structural interaction of NHE1, NCX1 and CaM proteins. The protein expression levels of NHE1, NCX1 and CaM in native human HCC tissues were markedly higher than those in normal liver tissues. IL6 upregulated the expressions of NHE1, NCX1 and CaM in Huh7 and HepG2 cells. NHE1, NCX1 and CaM mediated the promotion of IL6 on the proliferation, migration and invasion of Huh7 and HepG2 cells and the growth of HCC in nude mice. In conclusion, IL6 activates the functional activity of NHE1 and induces the functional and structural interaction of NHE1, NCX1 and CaM. The interaction of NHE1, NCX1 and CaM mediates the effects of IL6 on human HCC.

Growth and biochemical composition of filamentous microalgae Tribonema sp. as potential biofuel feedstock.

Filamentous oleaginous microalgae Tribonema minus have advantages in relatively easy harvesting and grazers resistance in mass cultivation due to its filaments in previous study. To evaluate whether the genus Tribonema is a valuable candidate for use in biofuel production, the morphology, growth, biochemical composition and fatty acid profile of six filamentous microalgae strains Tribonema sp. were investigated. All the strains are unbranched filament in single row of elongated cylinder, attaining 0.5-3 mm in length. The growth rates of tested strains were 0.35-0.42 g L(-1) d(-1). Generally, for all strains, decrease in protein content was followed by a slight increase in lipid and significant increase in carbohydrate in early phase, afterwards, lipid increased constantly inversely to decrease in carbohydrate content. After 15-day cultivation, total lipid contents of tested strains ranged from 38-61 %, of which TAG were the majority and palmitic acid (C16:0) and palmitoleic acid (C16:1) were the dominant components. The study confirmed that the genus Tribonema is the potential for biodiesel and bioethanol production upon culture time.

The P2Y2 nucleotide receptor mediates the proliferation and migration of human hepatocellular carcinoma cells induced by ATP.

ATP is an abundant biochemical component of the tumor microenvironment and a physiologic ligand for the P2Y2 nucleotide receptor (P2Y2R). In this study, we investigated the effect of ATP on the cellular behavior of human hepatocellular carcinoma (HCC) cells and the role of P2Y2R in ATP action and aimed to find a new therapeutic target against HCC. The experiments were performed in native isolated human HCC cells, normal hepatocytes, human HCC cell lines, and nude mice. We found that the mRNA and protein expression levels of P2Y2R in native human HCC cells and the human HCC cell lines HepG2 and BEL-7404 were enhanced markedly compared with human normal hepatocytes and the normal hepatocyte line LO2, respectively. ATP induced intracellular Ca(2+) increases in HCC cells and promoted the proliferation and migration of HCC cells and the growth of HCC in nude mice. The P2Y receptor antagonist suramin, P2Y2R-specific shRNA, the store-operated calcium channel inhibitors 2-aminoethoxydiphenyl borate (2-APB) and 1-(ß-3-(4-methoxy-phenyl) propoxyl-4-methoxyphenethyl)1H-imidazole-hydrochloride (SKF96365), and stromal interaction molecule (STIM1)-specific shRNA inhibited the action of ATP on HCC cells. In conclusion, P2Y2R mediated the action of ATP on the cellular behavior of HCC cells through store-operated calcium channel-mediated Ca(2+) signaling, and targeting P2Y2R may be a promising therapeutic strategy against human HCC.

A rapid thin-layer chromatography bioautographic method for detecting the monoamine oxidase inhibitors in plants.

It is well known that the isolation of monoamine oxidase (MAO) inhibitors from natural sources is an important strategy for drug development in the treatment of depression, Parkinson's disease and Alzheimer's disease. The present work describes developing a thin-layer chromatography (TLC) bioautographic method for detecting MAO inhibitors from plant extracts. The basic principle of the method is that the enzyme oxidises tryptamine into an aldehyde which in turn reacts with 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide to form a blue formazan which makes a blue-coloured background on the TLC plates. Inhibitors of MAO produced white spots on the background. The new TLC bioautographic method has several advantages such as lower consumption of the enzyme, shorter experimental time, more easily observed background of TLC plate and better reproducibility. The detection limits were 10 ng for several known MAO inhibitors.

Biofilm cultivation of the oleaginous microalgae Pseudochlorococcum sp.

The cultivation of microalgae in biofilm has been a potential way to overcome the shortcoming of conventional algal culture modes of open pond and photobioreactors in liquid suspension. However, the growth characteristics and related effect factors of the biofilm are still far from being understood. In this work, oleaginous microalgae species Pseudochlorococcum was cultured in an attached biofilm and influential factors on the growth rate of biofilm were investigated. The results showed that Pseudochlorococcum sp. preferred to accumulate more biomass on hydrophilic substrata than on hydrophobic one. The photon flux density of 100 µmol m(-2 )s(-1) was its light saturation point. The optimal inoculum density was about 3-5 g m(-2). The appropriate concentrations of nitrogen, phosphorus in medium and CO(2) in aerated gas were determined as 8.8, 0.22 mmol L(-1) and 1 %, respectively.

The growth, lipid and hydrocarbon production of Botryococcus braunii with attached cultivation.

The green alga Botryococcus braunii is regarded as a potential source of renewable fuel due to its high lipid and hydrocarbon contents. However, the slow growth rate damaged its feasibility for biofuel production. In this study, a novel method of 'attached cultivation' was introduced to incubate B. braunii FACHB 357 (B race). A high biomass productivity of 6.5 gm(-2)d(-1) was achieved in single layer attached system at early stage of cultivation. At day 10, the biomass, lipid and hydrocarbon productivities were 5.5, 2.34 and 1.06 gm(-2)d(-1), respectively. Under nitrogen starvation condition, both of the contents of lipid and hydrocarbon were increased, whereas the profile of hydrocarbon kept almost unchanged, while the content for oleic acid (18:1) increased and linolenic acid (18:3) decreased. With a multi-layer photobioreactor, a biomass productivity of 49.1 gm(-2)d(-1) or a photosynthetic efficiency of 14.9% (visible light) were obtained under continuous illumination of 500 µmolm(-2)s(-1).

Attached cultivation technology of microalgae for efficient biomass feedstock production.

The potential of microalgae biofuel has not been realized because of low productivity and high costs associated with the current cultivation systems. In this paper, an attached cultivation method was introduced, in which microalgae cells grew on the surface of vertical artificial supporting material to form algal film. Multiple of the algal films were assembled in an array fashion to dilute solar irradiation to facilitate high photosynthetic efficiency. Results showed that a broad range of microalgae species can grow with this attached method. A biomass productivity of 50-80 g m(-2) d(-1) was obtained outdoors for Scenedesmus obliquus, corresponding to the photosynthetic efficiency of 5.2-8.3% (total solar radiation). This attached method also offers lots of possible advantages over traditional open ponds, such as on water saving, harvesting, contamination controlling and scale-up. The attached cultivation represents a promising technology for economically viable production of microalgae biofuels.

Expression and functional role of vacuolar H(+)-ATPase in human hepatocellular carcinoma.

Tumor cells often exist in a hypoxic microenvironment, which produces acidic metabolites. To survive in this harsh environment, tumor cells must exhibit a dynamic cytosolic pH regulatory system. Vacuolar H(+)-adenosine triphosphatase (V-ATPase) is considered to play an important role in the regulation of the acidic microenvironment of some tumors. In this study, we made an investigation on the expression and functional role of V-ATPase in native human hepatocellular carcinoma (HCC). The results showed that the messenger RNA and protein expression levels of V-ATPase subunit ATP6L in native human HCC tissues were markedly increased, compared with normal liver tissues. Immunohistochemical analysis further confirmed the enhanced expression of V-ATPase ATP6L in human HCC cells and revealed that V-ATPase ATP6L was distributed in the cytoplasm and plasma membrane of HCC cells. The results from immunofluorescence and biotinylation of cell surface protein showed that V-ATPase ATP6L was conspicuously located in the plasma membrane of human HCC cells. Bafilomycin A1, a specific V-ATPase inhibitor, markedly slowed the intracellular pH (pHi) recovery after acid load in human HCC cells and retarded the growth of human HCC in orthotopic xenograft model. These results demonstrated that V-ATPase is up-regulated in human HCC and involved in the regulation of pHi of human HCC cells. The inhibition of V-ATPase can effectively retard the growth of HCC, indicating that V-ATPase may play an important role in the development and progression of human HCC, and targeting V-ATPase may be a promising therapeutic strategy against human HCC.

Evaluating human liver reserve function by measuring serum concentrations of phenacetin and its metabolites.

OBJECTIVES: To evaluate human liver reserve function (LRF) by a simple and efficient method for measuring serum concentrations of phenacetin and its metabolites. METHODS: Overall 20 patients with liver cirrhosis (Child-Pugh score ≥ 7, aged 48-79 years), 30 healthy young volunteers (aged 18-40 years), and 20 healthy elderly volunteers (aged 61-80 years) were enrolled. All participants received a single oral dose of 0.5 g phenacetin. Liquid chromatography tandem mass spectrometry was used to determine the serum concentrations of phenacetin and its metabolites, including acetaminophen, acetaminophen glucuronide and acetaminophen sulfate. RESULTS: The serum concentration of phenacetin was significantly higher in cirrhotic patients than those in either of the healthy volunteer groups (P < 0.001). It was higher in healthy elderly volunteers than that in healthy young ones but there was no statistically significant difference (P > 0.05) between them. The serum concentrations of acetaminophen, acetaminophen glucuronide and acetaminophen sulfate were significantly lower in cirrhotic patients than in the healthy controls (P < 0.001). The serum concentrations of these three metabolites in healthy elderly volunteers were lower than those in healthy younger volunteers but again, there was no statistical significant difference (P > 0.05). The serum concentration of acetaminophen in healthy male volunteers was significantly higher than that in the women (P < 0.05). CONCLUSION: Monitoring cytochrome P450 1A2 (CYP450 1A2)-mediated phenacetin metabolism is a simple and efficient method for evaluating human LRF. This method would warrant further validation in a large cohort clinical study.

BaNbO(IO3)5: a new polar material with a very large SHG response.

By combination of Nb(5+) (having a d(0) electronic configuration) and the lone-pair-containing iodate anion, a new SHG material, BaNbO(IO(3))(5), has been prepared. It exhibits a very large SHG response (approximately 14 times that of KH(2)PO(4) and approximately 660 times that of alpha-SiO(2)) and is phase-matchable. The material has high thermal stability and a wide transparent region.