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Trimethylamine N-oxide (TMAO) and phenylacetylglutamine (PAGln) are associated with acute myocardial infarction (AMI) and type 2 diabetes mellitus (T2DM). This study developed and validated a simple method firstly for simultaneously quantifying serum TMAO and PAGln using liquid chromatography-tandem mass spectrometry (LC-MS/MS).Serum samples from patients with T2DM, AMI, and healthy subjects were analyzed using a new LC-MS/MS method to evaluate TMAO and PAGln levels. Statistical analyses were performed to evaluate TMAO and PAGln distributions among different groups.Retention and separation of the two metabolites were achieved within 5 min. For both analytes, the assay was linear in a 0.02-10 µg/mL range, with 0.99 average linear correlation coefficients, and quantification limit values of approximately 0.010 µg/mL. The average recoveries of TMAO and PAGln were 96.3 % and 96.4 %, respectively. The intra-run and total coefficient variations were 3.5-4.8 % and 3.9-5.7 % respectively for TMAO; and 4.0-5.1 % and 4.6-6.3 % respectively for PAGln. TMAO and PAGln showed a moderate correlation (P 0.001) and their levels in patients with T2DM were significantly higher than those in healthy individuals (P 0.001). TMAO levels were higher in patients with T2DM than in patients with AMI (P 0.01). Patients with AMI had higher PAGln levels than healthy individuals (P 0.05). After adjusting for sex and age, the top tertile of PAGln was positively correlated with T2DM and AMI while that of TMAO was positively correlated with T2DM.Overall, a robust isotope dilution LC-MS/MS method was established, which may be beneficial for assessing the association between two metabolites with AMI and T2DM.

Glioma is a common type of malignancy in the central nervous system. The pathogenesis of glioma is complex and the underlying mechanisms remain largely unknown. In our study, exosomes were exacted from patient samples, and the isolated exosomes were confirmed by transmission electron microscope. The expression of circRNA_104948, miR-29b-3p and DNMT3B were determined using RT-qPCR. Proliferative activity of cell was examined using CCK-8 assay. Cell apoptotic rate was evaluated by flow cytometry. The expression levels of proliferation or apop-tosis markers were determined using western blotting. Our data suggested that circRNA_104948 was upregulated in plasma exosomes/tissue samples of glioma patients and glioma cell lines. Furthermore, cell proliferation was enhanced and apoptosis was suppressed in normal astrocytes treated with exosomal circRNA_104948, and the effects were reversed by sh-circRNA_104948. In addition, miR-29b-3p is a novel target of circRNA_104948, and DNMT3B is a putative downstream molecule of miR-29b-3p. circRNA_104948 could regulate the proliferation/apoptosis of astrocytes through miR-29b-3p/DNMT3B/MTSS1 signaling, and the biological behavior changes induced by glioma-Exo were reversed by miR-29b-3p mimics; upregulated cell growth caused by miR-29b-3p inhibitors was abrogated by the knockdown of DNMT3B; the effects induced by miR-29b-3p mimics were abolished by the overexpression of DNMT3B. Our findings revealed the important roles of circRNA_104948 on the development of glioma, and circRNA_104948/miR-29b-3p/MTSS1/DNMT3B pathway may be a potential candidate for the target therapy of glioma patients.

Gastric cancer with muscle weakness and pain as the first manifestation is rare. This paper reports the clinical features of a case of gastric cancer misdiagnosed as polymyositis, and to differentiate from similar diseases. Treatment remedies: Admission examination showed fever, muscle strength decline, muscle tenderness, muscle enzyme significantly increased. Polymyositis was first diagnosed, but the patient had significant decrease in whole blood cells. Combined with the examination results, it was found that the disease involved respiratory system, circulatory system, blood system, nervous system, urinary system and other systems, and symptomatic and supportive treatment was given. Post treatment evaluating: The symptoms of patients with progressive aggravation, not in line with the clinical outcome of polymyositis after treatment. After the general consultation of the whole hospital, bone marrow puncture and abdominal CT examination were completed, and the final diagnosis was extensive metastasis of gastric cancer.Conclusions: The early symptoms of gastric cancer are atypical, leading to misdiagnosis. This paper reports the clinical features of this case and differentiates it from similar diseases, so as to accumulate clinical data for the diagnosis of the disease and call for early gastric cancer screening.

Stroke is a significant cardiovascular disease that influences the health of human beings all over the world, especially the elderly population. It is reported that the blood-brain barrier (BBB) can be easily destroyed by stroke, which is one of the main factors responsible for macrophage infiltration and central nervous inflammation. Here, we report the protective effects of Trelagliptin against BBB injury and macrophage infiltration. Our results indicate that the infraction volume, the neurological score, and macrophage infiltration staining with CD68 were increased in middle cerebral artery occlusion (MCAO) mice but significantly reversed by treatment with Trelagliptin. Additionally, Trelagliptin reduced the permeability of the BBB by increasing the expression of the tight junction zonula occludens protein-1 (ZO-1) in the cerebral cortex. In an in vitro hypoxia model of endothelial cells, the increased migration of macrophages, enlarged permeability of endothelial monolayer, downregulation of ZO-1, and elevated expression level of CXCL1 by hypoxic conditions were all reversed by treatment with Trelagliptin in a dose-dependent manner. Our results demonstrate that Trelagliptin might mitigate macrophage infiltration by preventing the breakdown of the blood-brain barrier in the brains of MCAO mice.

Background Stroke serves as a prevalent cerebrovascular disorder with severe cerebral ischemia/reperfusion (CIR) injury, in which neural stem cells (NSCs) play critical roles in the recovery of cerebral function. Circular RNAs (circRNAs) have been widely found to participate in stroke and NSC modulation. However, the role of circRNA TTC3 (circTTC3) in the regulation of CIR injury and NSCs remains elusive. Here, we aimed to explore the impact of circTTC3 on CIR injury and NSCs. Methods The middle cerebral artery occlusion/repression (MCAO/R) model was established in C57BL/6J mice. The primary astrocytes were isolated from the cerebellum from C57BL/6J mice. The primary NSCs were obtained from rat embryos. The effect of circTTC3 on CIR injury and NSCs was analyzed by TTC staining, qPCR, Western blot, LDH colorimetric kits, MTT assays, Annexin V-FITC Apoptosis Detection Kit, luciferase reporter gene assays, and others in the system. Results Significantly, the expression of circTTC3 was elevated in the MCAO/R mice and oxygen and glucose deprivation (OGD)-treated astrocytes. The depletion of circTTC3 attenuated cerebral infarction, neurological score, and brain water content. The OGD treatment induced apoptosis and the levels of lactate dehydrogenase (LDH) in the astrocytes, in which circTTC3 depletion reduced this phenotype in the system. Moreover, the depletion of circTTC3 promoted the proliferation and upregulated the nestin and β-tubulin III expression in NSCs. Mechanically, circTTC3 was able to sponge miR-372-3p, and miR-372-3p can target Toll-like receptor 4 (TLR4) in NSCs. The miR-372-3p inhibitor or TLR4 overexpression could reverse circTTC3 depletion-mediated astrocyte OGD injury and NSC regulation. Conclusion Thus, we conclude that circTTC3 regulates CIR injury and NSCs by the miR-372-3p/TLR4 axis in cerebral infarction. Our finding presents new insight into the mechanism by which circTTC3 modulates CIR injury and NSC dysfunction. CircTTC3, miR-372-3p, and TLR4 may serve as potential targets for the treatment of CIR injury during stroke.

A 12-year-old castrated male dog with nonambulatory tetraplegia was diagnosed with spinal stenosis at C3-C4 through X-ray examination and with ventral extradural spinal compression at C3-C4 through myelography and computed tomography. The diagnosis of traditional Chinese veterinary medicine was local Qi and blood stagnation, spleen Qideficiency, blood deficiency, and kidney Yang deficiency. We initiated treatment using a combination of acupuncture and Chinese medicine iontophoresis with laser therapy. After 12 treatment days, there was a significant improvement in the dog's ambulation function, which was indicated by proper walking and flexible head-turning.This indicates that combining acupuncture and Chinese medicine iontophoresis could be a potential treatment for chronic progressive cervical intervertebral disk disease in dogs.Fallbericht: Bei einem 12-jährigen kastrierten Rüden mit Tetraplegie und Gehunfähigkeit wurde mittels Röntgenuntersuchung eine Spinalkanalstenose bei C3–C4 diagnostiziert und mittels Myelographie und Computertomographie eine ventrale extradurale Spinalkanalkompression bei C3–C4 nachgewiesen. Die Diagnose gemäß der traditionellen chinesischen Tiermedizin lautete lokale Qi- und Blutstagnation, Mangel an Milz-Qi, Blutmangel und ein Nieren-Yang-Mangel. Wir leiteten eine Kombinationstherapie mit Akupunktur und Iontophorese der chinesischen Medizin sowie Lasertherapie ein. Nach 12 Behandlungstagen war eine signifikante Verbesserung der Gehfunktion des Hundes zu beobachten, die sich durch regelrechtes Gehen und flexibles Drehen des Kopfes zeigte. Schlussfolgerung: Dies spricht dafür, dass die Kombination von Akupunktur und Iontophorese der chinesischen Medizin eine potenzielle Behandlungsmöglichkeit bei chronisch progredienter zervikaler Bandscheibenerkrankung bei Hunden sein könnte.

Background Hepatitis B virus (HBV) causes acute and chronic infection in the clinic. Hepatocellular carcinoma (HCC) is closely linked to HBV infection. Serum Golgi protein 73 (GP73) increases during HBV infection. However, the role of GP73 during HBV infection and the occurrence of HBV-related HCC is still poorly understood. Methods The underlying role of HBV-induced GP73 in regulating HCC development was investigated in this study. GP73 expression in HBV-related clinical HCC tissues and in HBV infected hepatoma cells and primary human hepatocytes were evaluated by immunohistochemistry, ELISA, Western Blotting and Quantitative real-time PCR (QRT-PCR) analysis. GP73 mediated tumorigenesis were detected by flow cytometry, QRT-PCR, xenograft nude mice and sphere formation assay. The effects of GP73 and HBV infection on host's innate immune responses in hepatocytes were further investigated by Western Blotting and Quantitative real-time PCR (QRT-PCR) analysis. Results Initially, we discovered that HBV-positive HCC tissues had significantly higher expression of GP73. Ectopic expression of the HBV gene could induce GP73 expression in primary human hepatocytes and hepatoma cells in vitro. In addition, we discovered that GP73 promotes HCC in both normal liver cells and hepatoma cells. We also found that ectopic expression of HBV genes increases GP73 expression, suppressing the host's innate immune responses in hepatocytes. Conclusions Our results demonstrate that HBV facilitates HCC development by activating GP73 to repress the host's innate immune response. This study adds to our understanding of the pathogenesis of HBV infection-induced HCC. The findings also provide preclinical support for GP73 as a potential HCC prevention or treatment target.

Bo Yang,1 Li-E Zang,1 Jing-Wen Cui,2 Ming-Yuan Zhang,2 Xue Ma,3 Lin-Lin Wei4 1Department of Neurology, The First Affiliated Hospital of Jinzhou Medical University, Jinzhou, Liaoning, People’s Republic of China; 2Department of Neurosurgery, The First Affiliated Hospital of Jinzhou Medical University, Jinzhou, Liaoning, People’s Republic of China; 3Department of Emergency, The First Affiliated Hospital of Jinzhou Medical University, Jinzhou, Liaoning, People’s Republic of China; 4Department of Gynaecology, The First Affiliated Hospital of Jinzhou Medical University, Jinzhou, Liaoning, People’s Republic of ChinaCorrespondence: Lin-Lin WeiDepartment of Gynaecology, The First Affiliated Hospital of Jinzhou Medical University, No. 2, Section 5, Renmin Street, Jinzhou, Liaoning 121000, People’s Republic of ChinaTel +86-416-4605293Email linlinweii@hotmail.comBackground: Melatonin (MT) has potential protective effect on cerebral ischemia-reperfusion injury (CIRI), but its underlying regulatory mechanism has not been identified.Purpose: This study aimed to explore the role of miR-26a-5p-neuron-restrictive silencing factor (NRSF/REST), Janus kinase-2 (JAK2)-signal transducer and activator of transcription-3 (STAT3) pathway in the protection mechanism of MT against CIRI in vivo and in vitro.Methods: Sprague Dawley rats were induced with ischemia-reperfusion (IR) in vivo model; PC12 cells were induced with oxygen-glucose deprivation/reperfusion (OGD/R) in vitro model; and MT intervention was conducted before the model was established. The effect of MT on autophagy factors (LC3II/LC3I, P62), inflammatory factors (TNF-α, IL-6, IL-10) and oxidative stress indexes (MDA, GSHPx, SOD) was explored, and then the above three indexes were determined by real-time quantitative PCR, ELISA, and detection kit corresponding to oxidative stress indexes. The neuroprotective effect of MT pretreatment on brain IR injury was evaluated by neurological deficit scores and TUNEL method. The levels of miR-26a-5p and NRSF were detected by real-time quantitative PCR and Western blot, and the interaction between them was evaluated by dual luciferase report. The role of JAK2-STAT3 pathway in MT protection mechanism was verified by pathway blocker (AG490) and Western blot.Results: MT pretreatment can significantly reduce neurological deficit score and neuronal apoptosis, inhibit CIRI autophagy, inflammation and oxidative stress in vivo and in vitro, reduce LC3II/LC3I, TNF-α, IL-6, MDA and increase P62, IL-10, GSHPx, SOD. Further analysis identifies that downregulating miR-26a-5p or upregulating NRSF can eliminate the protective effect of MT, and NRSF is the direct target of miR-26a-5p. The protective effect of MT can also be eliminated under AG490 intervention.Conclusion: MT plays a protective role by regulating miR-26a-5p-NRSF and JAK2-STAT3 pathway to improve CIRI autophagy, inflammation and oxidative stress.Keywords: cerebral ischemia-reperfusion injury, melatonin, miR-26a-5p, NRSF, JAK2-STAT3

Threatened by the energy crisis and environmental pollution, most countries in the world are vigorously developing new energy vehicles to promote low-carbon environmental protection and boost a green transportation system. Based on the intelligent manufacturing standard system, this study constructed a new energy vehicle intelligent manufacturing development-influencing factor model. Taking the intelligent manufacturing development ability as the dependent variable, taking external environment factors, commonalities among the new energy vehicle enterprises, and industry progress as independent variables, five hypotheses are proposed. This study used a structural equation model to test the hypotheses and reveal the mechanism of how factors influence the new energy vehicle intelligent manufacturing. The results show that external environment factors and industry progress directly and positively affect the development capability of intelligent manufacturing of new energy vehicles, while the commonalities among the NEV enterprises have an indirect effect through industry progress on intelligent manufacturing of new energy vehicles. Based on the analysis, this study puts forward some suggestions for better development of new energy vehicle intelligent manufacturing.

Enterococcus faecalis is a Gram-positive facultative anaerobe involved in many fatal or refractory infections of humans. Silver, often used as silver ions (Ag+) or nanoparticles (AgNPs), is a strong and broad-spectrum antibacterial agent, but E. faecalis shows resistance against it. Despite this, the knowledge about the resistance of E. faecalis against silver is still lacking. In this study, the silver-resistant E. faecalis strains (AgR and ANR E. faecalis) were established through a serial selection method. Their biological and silver-resistant features as well as the Gene Ontology (GO) in comparison with the original E. faecalis were evaluated. The results showed that the silver-resistant E. faecalis could proliferate as original bacteria and had strong resistance against both Ag+ and AgNPs. The minimum bactericidal concentrations (MBCs) of AgNO3 on original, AgR, and ANR E. faecalis were 400 mg/L, 600 mg/L, and 500 mg/L, and the MBCs of AgNPs on these strains were 80 mg/L, 110 mg/L, and 130 mg/L, respectively. GO analysis revealed significant difference (P

The potential of probiotics for treating ulcerative colitis (UC) has attracted increasing attention. However, more studies are still needed to guide physicians on the proper selection and use of probiotics. Here, we propose that combination of multiple probiotics with different functions can reduce intestinal inflammation. In this study, the effects of probiotics (Lactobacillus reuteri, Bacillus coagulans, Bifidobacterium longum, and Clostridium butyricum) on the physiology and histopathology of colon were evaluated in a dextran sulfate sodium (DSS)-induced colitis mouse model. The combined species, as well as the species individually, were tested and compared with sulfasalazine (SASP) and two Chinese herbal therapies. Results show that the functions of the four probiotic strains were different in regulating intestinal immunity and barrier function. The four-species probiotic cocktail was more effective than the species individually and anti-inflammatory drugs in repairing the dysbiosis of mucosal microbial ecology and reducing intestinal inflammation. The multi-strain probiotic mixture increased the proportion of beneficial bacteria and decreased the proportion of pro-inflammatory bacteria in the colonic mucosa. In addition, probiotic mixture significantly enhanced the expression of IL-10 and intestinal barrier function. These results suggest that a combination of multiple probiotics with different functions has synergistic effects and can restore the balance of interactions between microorganisms and immunological niches.

As one of the conventional treatment methods, acupuncture is an indispensable component of Traditional Chinese Medicine. Currently, acupuncture has been partly accepted throughout the world, but the mechanism of acupuncture is still unclear. Since the theory of the neuro-endocrine-immune network was put forward, new insights have been brought into the understanding of the mechanism of acupuncture. Studies have proven that acupuncture is a mechanical stimulus that can activate local cell functions and neuroreceptors. It also regulates the release of related biomolecules (peptide hormones, lipid hormones, neuromodulators and neurotransmitters, and other small and large biomolecules) in the microenvironment, where they can affect each other and further activate the neuroendocrine-immune network to achieve holistic regulation. Recently, growing efforts have been made in the research on the mechanism of acupuncture. Some researchers have transitioned from studying the mechanism of acupuncture as a single linear pathway to using systems approaches, including metabolomics, genomics, proteomics and biological pathway analysis. This review summarizes the research progress on the neuro-endocrine-immune network related mechanism of acupuncture and discusses its current challenges and future directions.

This work explores the use of water-in-supercritical carbon dioxide (scCO2) microemulsions for the extraction of polar metabolites from plants. Stevia rebaudiana Bertoni leaves and polyethylene glycol trimethylnonyl ether (TMN) surfactants were selected for a case study. A CO2-water-TMN 10 mixture at 35 °C and 30.0 MPa extracted 7 mg target analyte/g dry leaves. The extraction was proven to occur due to a water-surfactant liquid solution rather than a water-in-CO2 microemulsion. Using a modified extraction setup, the microemulsion was created prior to extraction. TMN 6 was able to dissolve enough water in CO2 to extract steviol glycosides, in detectable but very small amounts.

CO2-expanded liquids (CXLs) is a class of solvent systems giving relatively low viscosity in comparison to neat organic solvents. The challenge is to achieve a high overall polarity of the CXL, despite the presence of compressed liquid CO2. In this study, the relative static permittivity (epsilon(r)) was measured for binary and ternary one-phase mixtures of CO2 + green solvent using an in-line microfluidic device. In addition, Kamlet-Taft solvatochromic parameters were experimentally determined, allowing the characterization of the polarizability, acidity, and basicity. Novel data is shown for binary and ternary systems of CO2 expanded ethanol and ethyl lactate, with or without glycerol or water added, at moderate conditions of pressure and temperature (8 MPa and 35 degrees C). One-phase CXLs systems of a wide relative permittivity range (7.7-75.3) at lowered viscosity were enabled, by mixing different green solvents with CO2.

Enterococcus faecalis (E. faecalis) is a Gram-positive bacterium closely related to many refractory infections of human and shows the resistant ability against the antibacterial effects of silver. Simvastatin is a semisynthetic compound derived from lovastatin and a hydroxymethyl glutaryl coenzyme A(HMG-COA) reductase inhibitor showing certain inhibitive effects on bacteria. The main purpose of this study was to establish and characterize the Ag+/silver nanoparticles (AgNPs)-resistant E. faecalis, and further evaluate the function of extracellular polymeric substances (EPS) in the silver resistance and the effect of simvastatin on the silver-resistance of E. faecalis. The results showed that the established silver-resistant E. faecalis had strong resistance against both Ag+ and AgNPs and simvastatin could decrease the silver-resistance of both original and Ag+/AgNPs-resistant E. faecalis. The Transmission electron microscopy (TEM), High-angle annular dark-field (HAADF) and mapping images showed that the silver ions or particles aggregated and confined in the EPS on surface areas of the cell membrane when the silver-resistant E. faecalis were incubated with Ag+ or AgNPs. When the simvastatin was added, the silver element was not confined in the EPS and entered the bacteria. These findings may indicate that the silver resistance of E. faecalis was derived from the entrapping function of EPS, but simvastatin could compromise the function of EPS to decrease the silver resistant ability of E. faecalis.

In present work, a polysaccharide (GCP-WS) was successfully produced by Chaetomium globosum CGMCC 6882 from wheat straw. Monosaccharide composition of GCP-WS was rhamnose, glucosamine, galactose, glucose, xylose, fructose and glucuronic acid in a molar ratio of 21.46:1.58:1.11:55.15:36.37:7.04:7.34. Meanwhile, molecular weight of GCP-WS was 3.538 × 104 Da with a polydispersity of 1.043. Antioxidant assays revealed GCP-WS had strong scavenging effects on DPPH radical, ABTS radical, superoxide anions and hydroxyl radical in a dose-dependent manner. Results displayed that at the concentration of 5 mg/mL, the corresponding scavenging abilities of GCP-WS reached 83.08 ± 2.58%, 85.69 ± 1.62%, 79.38 ± 2.03% and 81.39 ± 1.47%, respectively. GCP-WS shows the potential for development as an effective antioxidant agent.

As the most abundant macromolecules in nature, lignocelluloses are served as a promising and renewable source for sustainable production of high value chemical compounds. In present work, extrusion pretreatment with 23% (w/w) distilled water, 2% (w/w) glycerol and 1 g/L NaHCO3 as moisture agent, not only reduced the particle size, crystallinity and component contents (cellulose, hemicelluloses and lignin) of rice straw, but also effectively enhanced chaetoglobosin A yield and degradation rate of rice straw by C. globosum CGMCC 6882. Meanwhile, mycelial biomass of C. globosum CGMCC 6882 increased from 2.9 g/L to 7.0 g/L, mycelia growth time reduced by 2 days and chaetoglobosin A titer increased from 108.4 mg/L to 270.2 mg/L, representing an increase of 149.3%. Furthermore, degradation rate of rice straw by C. globosum CGMCC 6882 increased from 28.93% to 65.38%. This work provides a good guidance for production of chaetoglobosin A from lignocelluloses.

In present work, a polysaccharide (GCP) was produced by Chaetomium globosum CGMCC 6882 and characterized. GCP was composed of arabinose, galactose, glucose, xylose, mannose and glucuronic acid in a molar ratio of 0.64: 2.58: 23.53: 0.90: 2.47: 0.27 with molecular weight of 8.093 × 104 Da and polydispersity (Mw/Mn) of 1.014. Antibacterial characteristics and mechanism of GCP against Escherichia coli and Staphlococcus aureus were investigated by analysis of inhibition zones, minimum inhibitory concentration (MIC), alkaline phosphatase and β-galactosidase activities, electrical conductivity and bacterial morphology. Results showed that the MIC of GCP against E. coli and S. aureus were 1.75 mg/mL and 0.67 mg/mL, respectively. Moreover, GCP exerted antibacterial activities by disrupting the inner membrane and increasing the cell permeability, but had no influences on cell wall. This work indicated that GCP could be explored as a promising antibacterial agent in food and pharmaceutical industries.

The aim of this study was to develop a novel fermented soybean food (FSF) using selected Bacillus subtilis GD1, Bacillus subtilis N4, Bacillus velezensis GZ1, Lactobacillus delbrueckii subsp. bulgaricus and Hansenula anomala, as well as to assess its antioxidant and anti-fatigue activity. These Bacillus strains had excellent enzyme producing and soybean transformation capacity. FSF showed the highest peptide, total phenol, total flavonoid content, antioxidant activity, and suitable organic acid and biological amine content. In intense exercise mice, FSF treatment markedly increased hepatic glycogen level, decreased metabolite accumulation, improved the activities of antioxidant enzymes and decreased malondialdehyde (MDA) level in serum and liver, respectively. Furthermore, FSF treatment increased nuclear factor-erythroid 2-related factor 2 (Nrf2) and antioxidant response element (ARE)-dependent gene expression. Together, the selection of microbial starter culture and mixed culture fermentation are essential for the effective enrichment of bioactive compounds, and FSF has stronger antioxidant and anti-fatigue activity.

Aims To screen and identify safe micro-organisms used during Douchi fermentation, and verify the feasibility of producing high-quality Douchi using these identified micro-organisms. Methods and Results PCR-denaturing gradient gel electrophoresis (DGGE) and automatic amino-acid analyser were used to investigate the microbial diversity and free amino acids (FAAs) content of 10 commercial Douchi samples. The correlations between microbial communities and FAAs were analysed by statistical analysis. Ten strains with significant positive correlation were identified. Then an experiment on Douchi fermentation by identified strains was carried out, and the nutritional composition in Douchi was analysed. Results showed that FAAs and relative content of isoflavone aglycones in verification Douchi samples were generally higher than those in commercial Douchi samples. Conclusions Our study indicated that fungi, yeasts, Bacillus and lactic acid bacteria were the key players in Douchi fermentation, and with identified probiotic micro-organisms participating in fermentation, a higher quality Douchi product was produced. Significance and Impact of the Study This is the first report to analyse and confirm the key micro-organisms during Douchi fermentation by statistical analysis. This work proves fermentation micro-organisms to be the key influencing factor of Douchi quality, and demonstrates the feasibility of fermenting Douchi using identified starter micro-organisms.

In this work, paper-based microfluidic sampling was used together with potentiometric determination of ions (Cd2+, Cl−, Pb2+) and pH. Calibration solutions and samples were absorbed into paper substrates and potentiometric detection was performed by placing solid-state ion-selective electrodes (ISEs) and a reference electrode in direct contact with the paper substrate. The paper substrate was found to influence the determination of Pb2+ at low concentrations, indicating complexation of Pb2+ by chemically active groups present in paper. Paper-based microfluidic sampling was successfully applied in measurements of pH in various foodstuffs and environmental samples by employing a conventional pH glass electrode. However, some influence of the paper substrate on the measured pH was observed for samples with pH close to neutral. Despite some limitations, paper-based microfluidic sampling with potentiometric sensing opens new possibilities for direct chemical analysis of specific sample types. The method described in this work is particularly interesting for analysis of small sample volumes with a high content of solid impurities and for analysis of e.g. liquid or moisture from polluted surfaces and foodstuff.

Background: Titanium dioxide nanoparticles (TiO2 NPs) have been widely used in industry and daily life; their potential neurotoxic effects are of great concern. The aim of this study is to determine whether exposure to TiO2 NPs mediates neurotrophins and related receptor expression in the hippocampus of mice under TiO2 NPs-induced neuroinflammation. Methods: Mice were nasally administered with 0.25, 0.5, or 1 mg per kg body weight TiO2 NPs for nine months. How neurotrophin-related factors and signaling pathways might be affected by exposure to TiO2 NPs for nine months using real-time PCR and ELISA methods were investigated. Results: The results suggest that exposure to TiO2 NPs caused TiO2 NPs deposition, excessive proliferation of all glial cells and tissue necrosis in the hippocampus. The hippocampal injury due to TiO2 NPs exposure was closely associated with significant reduction in nerve growth factor, brain-derived neurotrophic factor, tyrosine kinases (Trk A, B), calcium/calmodulin-dependent protein kinases (CaMKII, CaMKIV), cyclic-AMP responsive element binding proteins (CREB-1, CREB-2), dopamine receptors (D1, D2) and Tyrosine hydroxylase expression in the hippocampus. Conclusions: The findings imply that long-term exposure to TiO2 NPs may induce neuroinflammation via impairing neurotrophin-mediated signaling pathways in animals.

Titanium dioxide nanoparticles (TiO2 NPs) have been used in environmental management, food, medicine, and industry. But TiO2 NPs have been demonstrated to cross the blood-brain barrier and store up in the brain organization, leading to glutamate-mediated neurotoxicity. However, the neurotoxicity in the brain is not well understood. In this study, mice were exposed to 1.25, 2.5, or 5 mg/kg body weight TiO2 NPs for 9 months, and the glutamate-glutamine cyclic pathway and expressions of glutamate receptors associated with the hippocampal neurotoxicity were investigated. Our findings showed elevations of glutamate release and phosphate-activated glutaminase activity, and reductions in glutamine and glutamine synthetase in the hippocampus following exposure to TiO2 NPs. Furthermore, TiO2 NPs significantly inhibited the expression of N-methyl-d-aspartate receptor subunits (including NR1, NR2A, and NR2B) and metabotropic glutamate receptor 2 in mouse hippocampus. These findings suggest that the imbalance of glutamate metabolism triggered inhibitions of glutamate receptor expression in the TiO2 NP-exposed hippocampus. © 2014 Wiley Periodicals, Inc. Environ Toxicol 31: 655-662, 2016.

Solvent injection has been given attention to enhance oil recovery by sole use or in combination with a thermal method to develop light, medium, and heavy-oil fields. To make this process efficient, retrieval of expensive solvent efficiently is required. This can be achieved by alternative injection of water (WAG) if the reservoir is homogeneous. In case of heterogeneous reservoirs (fractured carbonates or sands with wormholes), one needs to develop techniques other than viscous displacement to retrieve the solvent diffused into less permeable matrix portion. A method of injecting steam/hot water to heat the solvent to vaporize and retrieve it was introduced recently (steam-over-solvent injection in fractured reservoirs) and tested through core experiments. Although these tests provide valuable data to design the optimal temperature of injected water to make the process viable, the mechanics of the nucleation of the solvent vapor and its entrapment in the pores at the micro scale requires further research. A series of experiments using a 2-D etched glass micromodel (sandstone replica with a fracture) were carried out to investigate the mechanics of solvent retrieval and entrapment at variable temperature conditions. The micromodel was saturated with dyed processed oils and different solvents were injected through fracture. After the solvent was diffused into matrix completely to recover the oil, the model was heated mimicking a thermal method to reach the boiling point of the solvent and retrieve it. The wettability of the micromodel was also altered to achieve water-wet and oil-wet conditions as wettability dictates the phase distribution in the pores. Following the heating phase, water was injected to retrieve the remaining solvent in the liquid or vapor phase. Visual observations of solvent diffusion/dispersion into matrix and its retrieval from the matrix clarified the miscibility process in the presence of an immiscible phase, interaction between different phases in a complex heterogeneous system, and phase distributions as a function of temperature. This information can be used to determine the efficiency of solvent retrieval process and optimal application conditions for EOR applications in heterogeneous sands and carbonates.

Solvent injection has been given attention to enhance oil recovery by sole use or in combination with a thermal method to develop light and heavy-oil fields. To make this process efficient, one needs to retrieve the expensive solvent. This can be achieved by alternative injection of water if the reservoir is homogeneous. In case of heterogeneous reservoirs (fractured carbonates or sands with wormholes), one needs to develop techniques other than viscous displacement to retrieve the solvent diffused into less permeable matrix portion. A method of injecting low temperature steam/water to heat the solvent to vaporize and retrieve it was introduced recently. An alternative is to inject chemical solution to change the wettability and displace the matrix oil/solvent by capillary imbibition. Although it yields lower recoveries, injection of chemical solutions for the same purpose without pre-solvent treatment might be an efficient (more economical) EOR method. A series of 2-D etched glass micromodel (sandstone replica with a fracture) experiments were designed to investigate the mechanics of chemical injection with and without pre-solvent injection. Conventional surfactants (sulfonate series) as well as new generation chemicals (nanofluids, ionic liquids) were tested for this purpose. After testing and screening effective chemicals without pre-solvent injection, the same chemicals were used to retrieve the solvent and recovery additional oil for pre-solvent injected systems. The micromodel was saturated by dyed processed oils and a selected solvent was injected through the fracture. After the solvent was diffused into matrix completely to recover the oil in it, the model was heated mimicking a thermal method to reach the boiling point of the solvent and retrieve it. Following the heating phase, aqueous phase was injected to retrieve the remaining solvent in the liquid or vapor phase. Visual observations on chemical flooding process clarified the complex interactions among different phases considering small-scale heterogeneities. This information can help screen promising chemical and determine the efficiency of chemical injection with and without pre-solvent injection for EOR in heterogeneous sands and carbonates.