Association between gaming disorder and regional homogeneity in highly involved male adult gamers: A pilot resting-state fMRI study.

BACKGROUND: Gaming behavior can induce cerebral changes that may be related to the neurobiological features of gaming disorder (GD). Additionally, individuals with higher levels of depression or impulsivity are more likely to experience GD. Therefore, the present pilot study explored potential neurobiological correlates of GD in the context of depression and impulsivity, after accounting for video gaming behavior. METHODS: Using resting-state functional magnetic resonance imaging (fMRI), a cross-sectional study was conducted with 35 highly involved male adult gamers to examine potential associations between GD severity and regional homogeneity (ReHo) in the entire brain. A mediation model was used to test the role of ReHo in the possible links between depression/impulsivity and GD severity. RESULTS: Individuals with greater GD severity showed increased ReHo in the right Heschl's gyrus and decreased ReHo in the right hippocampus (rHip). Furthermore, depression and impulsivity were negatively correlated with ReHo in the rHip, respectively. More importantly, ReHo in the rHip was found to mediate the associations between depression/impulsivity and GD. CONCLUSIONS: These preliminary findings suggest that GD severity is related to ReHo in brain regions associated with learning/memory/mood and auditory function. Higher levels of depression or impulsivity may potentiate GD through the functional activity of the hippocampus. Our findings advance our understanding of the neurobiological differences behind GD symptoms in highly involved gamers.

[Determination of 39 fatty acids in liver of rats by gas chromatography-mass spectrometry].

Fatty acids not only form phospholipids that contribute to the formation of cell membranes but also participate in many metabolic activities, such as energy storage and cell signal transduction. The liver plays a key role in the synthesis and metabolism of fatty acids. The composition and contents of fatty acids in the liver are closely related to body health. Most fatty acid-detection methods require a large sample size and can detect only a small number of fatty acids. Therefore, a sensitive and efficient method to determine fatty acids in the liver is urgently required. Herein, a method based on gas chromatography-mass spectrometry (GC-MS) was established for the simultaneous determination of 39 fatty acids in 1.1 mg of liver tissue. Different extraction methods and derivatization conditions were compared to develop an optimal sample-treatment method. The performance of two different columns in separating the target fatty acids were also compared. A total of 10 mg of liver was added to 450 µL of normal saline and ground at -35 ℃ to obtain a homogenate. Next, 50 µL of the homogenate (equivalent to 1.1 mg of liver) was added with 750 µL of chloroform-methanol (1∶2, v/v) to extract total fatty acids. The fatty acid extracts were dried under nitrogen, and then derivatized at 100 ℃ for 90 min after being added with methanol containing 5% sulfuric acid. The fatty acid methyl esters were extracted with hexane and then separated on an SP-2560 capillary column (100 m×0.25 mm×0.2 µm; Supelco, USA) via GC-MS. The results revealed that all 39 fatty acid methyl esters detected had good linearities in the certain mass concentration ranges with correlation coefficients (R2) greater than 0.9940. The limits of detection (LOD) and quantification (LOQ) of these methyl esters in the liver were 2-272 ng/mg and 7-906 ng/mg, respectively. The accuracy and precision of the method were evaluated by spiking the liver homogenate with tridecylic acid and eicosanoic acid at low (0.09 µg/mg), moderate (0.90 µg/mg), and high (5.40 µg/mg) concentration levels. The recoveries ranged from 82.4% to 101.0% with an intraday relative standard deviations (RSDs) (n=5) of 3.2%-12.0% and interday RSDs (n=3) of 5.4%-13.4%. The method was successfully applied to detect fatty acids in the livers of four healthy male Sprague-Dawley (SD) rats and four male SD rats with abnormal liver function induced by perfluorooctane sulfonate (PFOS). PFOS is a persistent organic pollutant. Twenty-six fatty acids were detected in the livers of both groups. Among the fatty acids investigated, pentadecanoic acid (C15∶0), γ-linolenic acid (C18∶3n6), and elaidic acid (C18∶1n9t) cannot be detected by the methods reported in the literature. By contrast, the method developed in this study could separate the isomers of oleic acid (elaidic acid, C18∶1n9t; oleic acid, C18∶1n9c) and linolenic acid (linolelaidic acid, C18∶2n6t; linoleic acid, C18∶2n6c). In conclusion, the developed method is simple and can detect a large number of fatty acids using small sample amounts and few reagents. More importantly, it could successfully separate fatty acid isomers. These findings indicate that the developed method is suitable for the detection of fatty acid composition and contents in the liver in clinical and experimental research.

Solasonine induces apoptosis of the SGC-7901 human gastric cancer cell line in vitro via the mitochondria-mediated pathway.

Solasonine, a steroidal glycoalkaloid isolated from the herbal plant Solanum nigrum Linn., has shown active against multiple human cancers; however, there is little knowledge on the activity of solasonine against gastric cancer until now. This study aimed to examine the effect of solasonine on the biological behaviours of human gastric cancer SGC-7901 cells. The results showed that solasonine suppressed SGC-7901 cell proliferation in a dose-dependent manner. Solasonine treatment mainly induced the cell cycle arrest at G2 phase in SGC-7901 cells. Treatment with solasonine resulted in significant down-regulation of Bcl-2 and Caspase-3 protein expression and reduced Bax and Bcl-xL protein expression in SGC-7901 cells. Solasonine shows a comparable inhibitory effect on the proliferation of human gastric cancer SGC-7901 cells with cisplatin, and solasonine induces of SGC-7901 cell apoptosis through triggering the endoplasmic reticulum stress pathway and the mitochondrial pathway. Our data indicate that solasonine may be a promising agent for the treatment of gastric cancer.

Microbial community structure and co-occurrence are essential for methanogenesis and its contribution to phenanthrene degradation in paddy soil.

Although polycyclic aromatic hydrocarbons (PAHs) degradation under methanogenesis is an ideal approach to remediating PAH-polluted soil, the contribution of methanogenesis to soil PAH elimination and the relationships between microbial ecological characteristics and PAH degradation during this process remain unclear. Here, we conducted a short-term (60 days) incubation using a paddy soil amended with phenanthrene and examined the effects of a specific methanogenic inhibitor (2-bromoethanesulfonate, BES) on this process. As treatment assessments, the methane production activity (MPA), phenanthrene degradation rate (PDR), and microbial ecological characteristics were determined. The results indicated that BES significantly inhibited both soil MPA and PDR, and we detected a positive relationship between MPA and PDR. Furthermore, BES significantly altered the soil microbial community structure, and it was the microbial community structure but not α-diversity was significantly correlated with soil MPA and PDR. BES decentralized the co-occurrence of bacterial genera but intensified the co-occurrence of methanogens. Moreover, certain bacterial taxa, including Bacteroidetes-vadinHA17, Gemmatimonas, and Sporomusaceae, were responsible for the MPA and PDR in this paddy soil. Collectively, these findings confirm the role of methanogenesis in PAH elimination from paddy soil, and reveal the importance of microbial co-occurrence characteristics in the determination of soil MPA and pollutant metabolism.

Sleep disorders in Alzheimer's disease: the predictive roles and potential mechanisms.

Sleep disorders are common in patients with Alzheimer's disease, and can even occur in patients with amnestic mild cognitive impairment, which appears before Alzheimer's disease. Sleep disorders further impair cognitive function and accelerate the accumulation of amyloid-ß and tau in patients with Alzheimer's disease. At present, sleep disorders are considered as a risk factor for, and may be a predictor of, Alzheimer's disease development. Given that sleep disorders are encountered in other types of dementia and psychiatric conditions, sleep-related biomarkers to predict Alzheimer's disease need to have high specificity and sensitivity. Here, we summarize the major Alzheimer's disease-specific sleep changes, including abnormal non-rapid eye movement sleep, sleep fragmentation, and sleep-disordered breathing, and describe their ability to predict the onset of Alzheimer's disease at its earliest stages. Understanding the mechanisms underlying these sleep changes is also crucial if we are to clarify the role of sleep in Alzheimer's disease. This paper therefore explores some potential mechanisms that may contribute to sleep disorders, including dysregulation of the orexinergic, glutamatergic, and γ-aminobutyric acid systems and the circadian rhythm, together with amyloid-ß accumulation. This review could provide a theoretical basis for the development of drugs to treat Alzheimer's disease based on sleep disorders in future work.

Pharmacological Treatment of Vascular Dementia: A Molecular Mechanism Perspective.

Vascular dementia (VaD) is a neurodegenerative disease, with cognitive dysfunction attributable to cerebrovascular factors. At present, it is the second most frequently occurring type of dementia in older adults (after Alzheimer's disease). The underlying etiology of VaD has not been completely elucidated, which limits its management. Currently, there are no approved standard treatments for VaD. The drugs used in VaD are only suitable for symptomatic treatment and cannot prevent or reduce the occurrence and progression of VaD. This review summarizes the current status of pharmacological treatment for VaD, from the perspective of the molecular mechanisms specified in various pathogenic hypotheses, including oxidative stress, the central cholinergic system, neuroinflammation, neuronal apoptosis, and synaptic plasticity. As VaD is a chronic cerebrovascular disease with multifactorial etiology, combined therapy, targeting multiple pathophysiological factors, may be the future trend in VaD.

CircRNA_102179 promotes the proliferation, migration and invasion in non-small cell lung cancer cells by regulating miR-330-5p/HMGB3 axis.

Non-small cell lung cancer (NSCLC) accounting for 85 % of all lung cancer was one of the main causes of death worldwide. In this study, we investigated the role of circRNA_102179 in NSCLC development. The levels of circRNA_102179 in NSCLC tissues and cell lines were determined by quantitative real-time PCR assay (qRT-PCR). CCK8 and colony formation assays were applied to explore the effect of circRNA_102179 on the growth of NSCLC cells in vitro. Transwell assay was utilized to analyze the impact of circRNA_102179 on the migration and invasion of NSCLC cells. Target prediction and luciferase reporter assay were used to identify the interacting miRNA of circRNA_102179. The interaction among circRNA_102179/ miR-330-5p/HMGB3 was further validated by colony formation and Transwell invasion assays. Finally, the mouse xenograft NSCLC model was used to explore the role of circRNA_102179 in the tumor growth of NSCLC cells in vivo. CircRNA_102179 was overexpressed in NSCLC tissues and cells compared with normal lung tissues and human bronchial epithelial cells (HBEs). The down-regulation of circRNA_102179 markedly reduced the proliferation, migration, and invasion of NSCLC cells. Moreover, down-expression of circRNA_102179 significantly increased the level of miR-330-5p/HMGB3 in NSCLC cells. Further functional experiments indicated that over-expression of miR-330-5p reversed the inhibitory effect of circRNA_102179 on NSCLC cells growth, migration, and invasion. Our results reveal that circRNA_102179 facilitates the proliferation, migration, and invasion of NSCLC cell via modulating miR-330-5p/ HMGB3 axis in NSCLC cells.

Roles of pattern recognition receptors in diabetic nephropathy.

Diabetic nephropathy (DN) is currently the most common complication of diabetes. It is considered to be one of the leading causes of end-stage renal disease (ESRD) and affects many diabetic patients. The pathogenesis of DN is extremely complex and has not yet been clarified; however, in recent years, increasing evidence has shown the important role of innate immunity in DN pathogenesis. Pattern recognition receptors (PRRs) are important components of the innate immune system and have a significant impact on the occurrence and development of DN. In this review, we classify PRRs into secretory, endocytic, and signal transduction PRRs according to the relationship between the PRRs and subcellular compartments. PRRs can recognize related pathogen-associated molecular patterns (PAMPs) and danger-associated molecular patterns (DAMPs), thus triggering a series of inflammatory responses, promoting renal fibrosis, and finally causing renal impairment. In this review, we describe the proposed role of each type of PRRs in the development and progression of DN.

The linkage between methane production activity and prokaryotic community structure in the soil within a shale gas field in China.

Soil methane generation mainly driven by soil prokaryotic microbes can be coupled with the degradation of petroleum hydrocarbons (PHCs); however, the relationship between prokaryotic community structure and methane production activity in soil with the potential risk of PHC contamination is seldom reported. In this study, 3 soil samples (CS-1 to CS-3) in the area nearby an exploratory gas well and 5 soil samples (DC-1 to DC-5) in a drill cutting dump area were obtained from the Fuling shale gas field (Chongqing City, China). Then, the prokaryotic community structure was examined by Illumina Miseq sequencing, and the linkage between soil methane production rate (MPR) and prokaryotic community composition was analyzed. The results indicated that 2 samples (DC-4 and DC-5) collected from the drill cutting dump area had significantly higher MPR than the other samples, and a significant and positive relationship (r = 0.44, P < 0.05) was found between soil MPR and soil organic matter (OM) content. The prokaryotic community composition in the sample (DC-5) with the highest MPR was different from those in the other samples, and soil OM and MPR were the major factors significantly correlated with the prokaryotic community structure in this soil. The samples (DC-4 and DC-5) with higher MPR had a higher relative abundance of Archaea and different archaeal community structures from the other samples, and the MPR was the sole factor significantly correlated with the archaeal genus composition in this soil. Therefore, both the prokaryotic and archaeal community structures are essential in the determination of soil MPR, and the bacterial genus of Saccharibacteria and the archaeal genus of Methanolobus might be the key contributors for methane generation in this soil from the shale gas field.

Intratumoral IL-12 Gene Therapy Inhibits Tumor Growth In A HCC-Hu-PBL-NOD/SCID Murine Model.

PURPOSE: This study aimed to evaluate the efficacy and safety of intratumoral IL-12 gene therapy in an HCC-hu-PBL-NOD/SCID mouse model. MATERIALS AND METHODS: The HCC murine model was generated in NOD/SCID mice, and mice with grafted tumors were injected intraperitoneally with 2 × 107 human peripheral blood lymphocytes 14 days after modeling. After 4 days, mice were randomly divided into the 9597/IL-12 group, the 9597/plasmid group and the PBS group. The changes of tumor volume were measured and mouse peripheral blood was sampled post-treatment for ELISA and CBA analyses, and the grafted tumors were collected 28 days post-treatment for immunohistochemistry, ELISA, CBA and detection of cell cycle and apoptosis. RESULTS: The tumor volume was smaller in the 9597/IL-12 group than in the 9597/plasmid and PBS groups on days 7, 14, 21, and 28 post-treatment (P < 0.05). Higher IL-12 levels were detected in the peripheral blood and the supernatants of grafted tumor homogenates in the 9597/IL-12 group than in the 9597/plasmid and PBS groups 7, 14, 21 and 28 days post-treatment (P < 0.05). IHC revealed higher counts of CD3+T cells, CD4+T helper cells, IFN-γ Th1 cells+ and S-100 protein positive dentric cells and lower MVD in the 9597/IL-12 group than in the 9597/plasmid and PBS groups (P < 0.05). Flow cytometry showed a significantly higher proportion of HCC cells at the G0/G1 phase and a significantly lower proportion of HCC cells at the S phase in the 9597/IL-12 group than in the PBS group (P < 0.05) and a greater apoptotic rate of HCC cells in the 9597/IL-12 group than in the 9597/plasmid and PBS groups (P < 0.05). CONCLUSION: Intratumoral IL-12 gene therapy may inhibit tumorigenesis with mild adverse effects in a HCC-hu-PBL-NOD/SCID murine model through inhibiting angiogenesis, arresting cells in G0/G1 phase and inducing apoptosis.

[Investigation of the Coupling Mechanism Between Naphthalene Degradation and Denitrification of a Naphthalene Degraded Bacterial Consortium Under Denitrification].

To investigate the coupling mechanism between naphthalene degradation and denitrification using a liquid media containing naphthalene (sole carbon source) and nitrate, a naphthalene degradation bacterial consortium under denitrification was enriched from a soil with potential risk of PAH (polycyclic aromatic hydrocarbon) contamination. The bacterial community composition of the enriched consortium was analyzed by Illumina MiSeq Sequencing. Subsquently, the enriched consortium was cultured under anaerobic conditions for 9 days, and the concentrations of electron acceptors (nitrate and nitrite) for denitrification, gaseous reduction products (N2O and N2) involved in denitrification, and abundances of denitrification concerned genes (narG:periplasmic nitrate reeducates gene; nirS:cd1-nitrite reductase gene) were detected at days 1, 3, 7, and 9. The result of Illumina MiSeq Sequencing showed that Pseudomonas (Proteobacteria) was the most dominant genus in this enriched consortium. Under anaerobic conditions, the naphthalene removal rate of this enriched consortium was 49.11% within 9 days. Relative higher naphthalene degradation rates were found both at the beginning (day 1-3) and the end (day 7-9) of incubation, and these were significantly higher than at the middle (day 3-7) of the incubation stage (P<0.05). The concentration of nitrate decreased during the whole culture period, while the concentration of nitrite increased during the initial incubation (day 1-3) and rapidly decreased from day 3 to 9. Furthermore, obvious productions of N2O[3.39 µg ·(L ·h)-1] and N2[8.97 µg ·(L ·h)-1] were only measured at the end of incubation (day 7-9). The abundances of both narG and nirS increased during the incubation, indicating the continuous growth of denitrifiers in the enriched consortium during the incubation period. In summary, this study illustrated that both the nitrate reducing stage and gas producing steps of denitrification could be coupled with anaerobic naphthalene degradation, which might be helpful for a deeper investigation regarding the coupling mechanism between denitrification and anaerobic PAH degradation.

[Effect of Nitrate Amendment on Soil Denitrification Activity and Anthracene Anaerobic Degradation].

The degradation of soil polycyclic aromatic hydrocarbons (PAHs) under denitrification is one of the most important pathways for anaerobic PAH elimination, but little is known about the effect of nitrate (the terminal electron acceptor for denitrification) on soil denitrification activity and PAH degradation under anaerobic conditions. In this study, the effect of nitrate on soil anthracene anaerobic degradation and denitrification activity was investigated through an anaerobic microcosm experiment. Two groups of treatments without (N0) and with (N30) nitrate (30 mg·kg-1) amendment were conducted. Each group contained three treatments with different anthracene concentrations (0, 15, and 30 mg·kg-1, denoted as A0, A15, and A30, respectively). Therefore, a total of six treatments (N0A0, N0A15, N0A30, N30A0, N30A15, and N30A30) were incubated in darkness at 25℃ for 45 days, and the production rates of N2O and CO2, abundances of denitrification related genes (narG:periplasmic nitrate reductase gene; nirK:copper-containing nitrite reductase gene; and nirS:cd1-nitrite reductase gene), and soil anthracene content were measured at 3, 7, 14, and 45 days. The results indicated that the intensive denitrification enzyme activity in each treatment was only detected at day 3, which could be significantly enhanced by both nitrate and anthracene amendments. Subsequently, a sharp decline of denitrification enzyme activity was observed in each treatment, while anthracene showed an obvious inhibition of soil denitrification enzyme activity. The result of a two-way ANOVA also indicated that nitrate, anthracene, and their interactions had significant effects on soil denitrification enzyme activity. The result of a quantitative-PCR indicated that, during the incubation, the abundances of narG and nirS exhibited an increasing tendency, but the abundance of nirK was relatively constant compared with its former counterparts. The final removal rate of anthracene under anaerobic soil environment was in the range of 33.83%-55.01%, and neither the final removal rate nor the degradation rate of anthracene could be significantly affected by nitrate amendment during incubation. The anthracene degradation rates in the higher anthracene containing treatments (N0A30 and N30A30) were significantly higher than those in the lower anthracene containing treatments (N0A15 and N30A15). In summary, nitrate amendments had no effect on soil anthracene anaerobic degradation but could significantly affect soil denitrification enzyme activity and the abundance of denitrification related narG and nirS genes.

Effect of the nitrification inhibitor (3, 4-dimethylpyrazole phosphate) on the activities and abundances of ammonia-oxidizers and denitrifiers in a phenanthrene polluted and waterlogged soil.

Through a 60-day microcosm incubation, the effect of 3, 4-dimethylpyrazole phosphate (DMPP) on the activities and abundances of ammonia-oxidizers and denitrifiers in phenanthrene-polluted soil was investigated. Five treatments were conducted for clean soil (CK), phenanthrene added (P), phenanthrene and DMPP added (PD), phenanthrene and urea added (PU), and phenanthrene, urea, and DMPP added (PUD) soils. The results indicate that the potential nitrification rate (PNR) in the P treatment was significantly higher than that in the PD treatment only on day 7, whereas the PNR in the PU treatment was significantly higher than that in the PUD treatment on each sampling day. The abundance of soil ammonia-oxidizing bacteria (AOB) in the PU treatment was significantly higher than that in the PUD treatment on each sampling day. Moreover, the abundance of AOB but rather than the ammonia-oxidizing archaea (AOA) had significantly positive correlation with soil PNR (P < 0.05). DMPP showed no obvious effect on the soil denitrification enzyme activity (DEA), which could have inhibited the abundances of denitrification-related narG, nirS, and nirK genes. The results of this study should provide a deeper understanding of the interaction between soil polycyclic aromatic hydrocarbons (PAH) contamination, ammonia oxidization, and denitrification, and offer valuable information for assessing the potential contribution of denitrification for soil PAH elimination.

[Effects of Wetland Types on Distribution of Soil Methylmercury Based on the Region of Nanweng River in the Greater Xing'an Mountains].

Methylmercury (MeHg) is the most toxic organic matter of all mercury (Hg) compounds. Its toxicity is far higher than that of inorganic Hg. Methylmercury can enter the human body through food, threatening human health. Based on the Nanweng River National Nature Reserve in the Greater Xing'an Mountains, the differences in the distribution of MeHg in soil samples collected from four types of wetlands, including medium swamp, low swamp, island forest, and forest soil, and the effect of the soil properties on the MeHg content were investigated in this study. The results show that:① the trends of the total Hg and MeHg levels are inconsistent. The order of the mean total Hg content is island forest (138.76 mg·kg-1±101.97 mg·kg-1) > forest soil (117.57 mg·kg-1±32.44 mg·kg-1) > low swamp (71.8 mg·kg-1±1.42 mg·kg-1) > median swamp (65.11 mg·kg-1±26.69 mg·kg-1), while the mean MeHg content is in the order of island forest (1.14 µg·kg-1±1.15 µg·kg-1) > medium swamp (0.87 µg·kg-1±1.06 µg·kg-1) > low swamp (0.28 µg·kg-1±0.06 µg·kg-1) > forest soil (0.1 µg·kg-1±0.05 µg·kg-1); ② the contents of MeHg in the island forest and medium swamp were relatively high and fluctuated dramatically, whereas the contents of MeHg in the low swamp and forest soil were lower and showed little change between each sampling point; ③ the longer the marsh wetland formation was, the higher was the MeHg content, that is, median swamp (0.87 µg·kg-1±1.06 µg·kg-1) > low swamp (0.28 µg·kg-1±0.06 µg·kg-1); the soil pH was positively correlated with MeHg (P<0.05); in the medium swamp area, the content of MeHg was positively related to the water content (WC), organic matter (OM%), carbon (C%), and total mercury (THg; P<0.05); in the island forest areas, the MeHg content showed a linear relationship with the soil pH and NH4+-N (P<0.05). This study reveals the distribution characteristics of MeHg and its influencing factors for different wetland ecosystems, providing data supporting the comprehensive evaluation of MeHg accumulation in wetland ecosystems.

Co-effects of pyrene and nitrate on the activity and abundance of soil denitrifiers under anaerobic condition.

It has previously been confirmed that polycyclic aromatic hydrocarbons (PAHs) could be degraded by soil microbes coupling with denitrification, but the relationships among soil denitrifiers, PAHs, and nitrate under obligate anaerobic condition are still unclear. Here, co-effects of pyrene and nitrate on the activity and abundance of soil denitrifiers were investigated through a 45-day incubation experiment. Two groups of soil treatments with (N30) and without (N0) nitrate (30 mg kg-1 dry soil) amendment were conducted, and each group contained three treatments with different pyrene concentrations (0, 30, and 60 mg kg-1 dry soil denoted as P0, P30, and P60, respectively). The pyrene content, abundances of denitrification concerning genes (narG, periplasmic nitrate reductase gene; nirS, cd 1-nitrite reductase gene; nirK, copper-containing nitrite reductase gene), and productions of N2O and CO2 were measured at day 3, 14, 28, and 45, and the bacterial community structures in four represented treatments (N0P0, N0P60, N30P0, and N30P60) were analyzed at day 45. The results indicated that the treatments with higher pyrene concentration had higher final pyrene removal rates than the treatments with lower pyrene concentration. Additionally, intensive emission of N2O was detected in all treatments only at day 3, but a continuous production of CO2 was measured in each treatment during the incubation. Nitrate amendment could enhance the activity of soil denitrifiers, and be helpful for soil microbes to sustain their activity. While pyrene seemed had no influence on the productions of N2O and CO2, and amendment with pyrene or nitrate both had no obvious effect on abundances of denitrification concerning genes. Furthermore, it was nitrate but not pyrene had an obvious influence on the community structure of soil bacteria. These results revealed that, under anaerobic condition, the activity and abundance of soil denitrifiers both were insensitive to pyrene, but nitrate could improve the activity of soil denitrfiers and induce the shifts in soil bacterial community structure.

Comparative Investigation of Bacterial, Fungal, and Archaeal Community Structures in Soils in a Typical Oilfield in Jianghan, China.

Agricultural soils in oilfields have high risk for polycyclic aromatic hydrocarbon (PAH) pollution. In this study, from the Jianghan Oilfield (Hubei Province, China) with a history of >50 years, 7 soil samples (OS-1 to OS-7) were collected. Subsequently, the bacterial, archaeal, and fungal community structures were investigated by Illumina MiSeq sequencing, and the relationship between microbial community structure and soil PAH content was analyzed. The results indicated that bacterial and archaeal Chao 1 indices showed a significantly negative relationship with soil PAH content, and only the bacterial Shannon index had a significantly negative relationship with soil PAH content. Moreover, the community structure of bacteria (r 2 = 0.9001, p = 0.013) showed a stronger correlation with PAH content than that of fungi (r 2 = 0.7357, p = 0.045), and no significant relationship was found between archaeal community structure (r 2 = 0.4553, p = 0.262) and soil PAH content. In addition, the relative greater abundances of some bacterial genus belonging to Actinobacteria (Mycobacterium and Micromonospora) and Proteobacteria (Pseudomonas, Lysobacter, Idiomarina, Oxalobacteraceae, and Massilia), fungal genus belonging to Ascomycota (Sordariales and Pleosporales), and archaeal phylum (Euryarchaeota) were detected in the soil samples (OS-3 and OS-5) with greater PAH content. In summary, soil PAHs showed an obvious influence and selectivity on the soil microbiota. Furthermore, compared with fungi and archaea, bacteria was more sensitive to soil PAH pollution, and the diversity indices and community structure of bacteria both might be suitable indicators for assessment of soil PAH stress on the soil ecosystem.

[Inducing Effect of Modified Cytokine Cocktail on Dendritic Cells].

OBJECTIVE: To investigate the inducing effect of 'modified' cytokine cocktail on the dendritic cell maturation and migration capability. METHODS: PBMNC were isolated from human peripheral blood stem cell (PBSC) by using density gradient centrifugation, the immature DC (imDC) were induced by using GM-CSF and IL-4 in vitro. Total A549 RNA was transfected into imDC by using electroporation, which was stimulated to matuation by the "gold standard" cytokine cocktail and "modified" cytokine cocktail, respectively. The expression of DC surface markers (CD11c, HLA-DR, CD80, CD83 and CD86) and chemokine receptor (CCR5, CCR7 and CXCR4) were detected by flow cytometry; the mRNA expression levels of DC chemokine receptor (CCR2, CCR5, CCR7, CXCR3 and CXCR4) and chemokine (CCL2, CCL3, CCL5, CCL19, CCL21, CXCL10 and CXCL12) were detected by RT-PCR. RESULTS: As compared with "gold standard cytokine cocktail", the "modified" cytokine cocktail-induced DC expressed higher levels of surface markers (CD11c, HLA-DR, CD80, CD83 and CD86), chemokine receptors (CXCR4) and chemokine (CCL2, CCL3, CCL5, CCL19, CCL21, CXCL10 and CXCL12). CONCLUSION: The "modified" cytokine cocktail can more effectively induce the DC maturation, enhace the migratory capability of DC and more generate the immunostimulatory DC, when compared with the "gold standard" cytokine cocktail effect.

Tumor MICA status predicts the efficacy of immunotherapy with cytokine-induced killer cells for patients with gastric cancer.

In this study, we determine the relationship between the expression of major histocompatibility complex class I chain-related gene A (MICA) in gastric cancer tumors after D2 gastrectomy and the clinical outcome of a CIK-containing adjuvant therapy. Ninety-five consecutive patients with gastric cancer after D2 gastrectomy who received adjuvant chemotherapy combined with CIK cell therapy were enrolled. The MICA expression of their tumors was determined by immunohistochemistry (IHC). High expression of MICA protein was documented by IHC in 38 of 95 tumor samples (40.0 %). The MICA status was significantly associated with the age and stage, p = 0.008 and 0.023, respectively. Analysis of NKG2D on in vitro expanded CIK cells showed that the percentages of NKG2D+ in CD3+/CD56+, CD3-/CD56+, and CD3+/CD8+ cells populations were 97.2 ± 1.4, 97.9 ± 1.8, and 95.6 ± 2.1 %, respectively. For patient with high MICA-expressing tumors, the median DFS and OS were longer than for the patients with tumors with low expression of MICA; 46.0 versus 41.0 months (p = 0.027), and 48.0 versus 42.0 months (p = 0.031), respectively. In a multivariate analysis, stage and MICA expression were independent prognostic factors for DFS and OS. Our findings show that adjuvant chemotherapy plus CIK therapy treatment is a promising modality for treating gastric cancer patients after D2 gastrectomy. Especially, those who have tumors with high expression of MICA were more likely to benefit from such a treatment strategy. Subsequent studies in clinical trial cohorts will be required to confirm the clinical utility of these markers.

[Effects of PAHs Pollution on the Community Structure of Denitrifiers in a Typical Oilfield].

Agricultural soils in the oilfields have the potential risk of PAHs (polycyclic aromatic hydrocarbons) pollution, and the denitrification process with nitrate as the terminal electron acceptor might be important for soil PAHs elimination under anaerobic condition. In this study, 9 soil samples listed as JH-1 to JH-9 were collected from the JiangHan oilfield with a history of more than 50 years. Using the functional genes (nirK: Cu-nitrite reductase gene; nirS: cd1-nitrite reductase gene) involved in denitrification as biomarkers, the community structure of soil denitrifiers was investigated by quantitative-PCR and T-RFLP (terminal-restriction fragment length polymorphism) combined with clone library, and the relationship between soil properties and community structure of soil denitrifers was discussed. The result indicated that the copy numbers of nirK were higher than those of nirS in all soil samples, and the lowest copy numbers of nirK and nirS were both detected in the JH-4 with the highest PAHs content. Meanwhile, the correlation analysis also showed a negative correlation between the copy numbers of those functional genes and soil PAHs content (nirK: R2=0.54, P<0.05; nirS: R2=0.58, P<0.05). Furthermore, the result of T-RFLP indicated that the nirK community structures in different soil samples varied significantly, which was obviously unique in the sample (JH-4) with the highest PAHs content. The subsequent RDA (redundancy analysis) also demonstrated that soil PAHs content as well as the available nitrogen and phosphorus belonged to the most important factors affecting the nirK community structure in this oilfield soil. Compared with nirK, little variation was shown about the nirS community structure among the soil samples. However, the abundance of nirS-harboring pseudomonas had a remarkably positive relation with the soil PAHs content, which indicated that pseudomonas, a well known bacterial genus with strong ability to degrade organic pollutants, might be an essential driver for PAHs degradation via denitrification process in this oilfield soil.