[Effect of Substrate Concentration on SAD Collaborative Nitrogen and Carbon Removal Efficiency in an ABR Reactor].

In order to solve the problem of declining total nitrogen (TN) removal caused by anaerobic ammonium oxidation (ANAMMOX) and the suppression of organic matter for ANAMMOX, the anaerobic baffled reactor (ABR), inoculating ANAMMOX sludge and anaerobic sludge from a municipal WWTP, was selected to construct system of ANAMMOX coupled denitrification (SAD) by the control of different substrate concentration. The SAD was constructed to study the effects of different influent substrates (COD, NO2--N, NH4+-N) on the performance of nitrogen and carbon removal in the coupled system and pollutant removal rules. The results showed that the coupling reaction was achieved in the ABR reactor and the inhibitory effect of organic compounds on anaerobic ammonium oxidation bacteria (AAOB) was relieved. When influent concentrations of COD, NO2--N, and NH4+-N were 260, 185, and 100 mg·L-1, respectively, which equates to a ratio of 2.6∶1.85∶1, the concentrations of these substances in the effluent decreased to 10, 1.0, and 0.9 mg·L-1, respectively. The TN removal rate reached 99%, hence stable system operation and ultra-low emissions of carbon and nitrogen pollutants were achieved. Under different conditions of substrate concentrations and ratios, the targeted pollutants were generally eliminated in the first compartment, in which the removal rate reached higher than 75%, and ANAMMOX held the dominant position in the SAD coupled system.

[Start-up and Characteristics of the Microbial Community Structure of ANAMMOX].

An anaerobic ammonium oxidation (ANAMMOX) reactor was successfully started up in 17 days, with the up-flow anaerobic sludge blanket (UASB) reactor being seeded with mixed anaerobic sludge from laboratory cultures with an ANAMMOX function and aerobic activated sludge from a municipal sewage treatment plant in a volume ratio of 1:2. The processes could be divided into two phases of hydrolysis, enhanced and steady. Anaerobic ammonium oxidation bacteria (AAOB) were enriched by improving the reactor volume load gradually after the steady phase. When the volume load increased from 0.10 kg·(m3·d)-1 to 0.44 kg·(m3·d)-1, the removal of total nitrogen (TN) also increased from 0.09 kg·(m3·d)-1 to 0.42 kg·(m3·d)-1. The color of the sludge changed from a light red that deepened gradually in the UASB reactor. At that time, the proportion of the sludge particle size greater than 0.2 mm increased from 10.90% to 38.37%.The sludges from the inoculation phase and from the phase when the volume load was increasing were analyzed by high-throughput sequencing, indicating that Chloroflexi, Proteobacteria, WWE3, Actinobacteria, Planctomycetes, and so on were the dominant species. The proportion of Proteobacteriain the denitrification bacteria was gradually reduced from 21.60% to 14.20% with an increase in the degree of AAOB enrichment, while the Planctomycetes increased from 0.73% to 15.50%. Candidatus Brocadia, Candidatus Jettenia, and Candidatus Kuenenia were the main species of Planctomyceteswhen the volume load increased to 0.44 kg·(m3·d)-1 in the reactor, and the Candidatus Brocadia was the main species of AAOB, which accounted for 13.40%.

[Characteristics of Microbial Community in Each Compartment of ABR ANAMMOX Reactor Based on High-throughput Sequencing].

In order to investigate the characteristics of microbial community in each compartment of ABR anammox reactor, a five-compartment ABR reactor was used to analyze the microbial community by Miseq High-throughput Sequencing during the steady operational process. The results indicated that the denitrifying bacteria coexisted in the reactor, such as Proteobacteria, Planctomycete, and Nitrospirae bacteria, and the percentages of these three microbial populations in the sludge were 11.66%-20.28%, 2.18%-7.94% and 0.19%-6.30%, respectively. In addition, there were four dominant genera in the phylum Proteobacteria:Rhodoplanes, Dok59, Rubrivivax and Bdellovibrio. Furthermore, Candidatus brocadia and Candidatus kuenenia were the main genera in the phylum Planctomycete. The color of sludge in the five compartments, in turn, varied from red to black. In addition, the biodiversity index of Chao, ACE, Shannon and Simpson indicated that the richness and diversity of microbial community increased gradually, and at the same time, the relative abundance of Proteobacteria increased while that of Planctomycetes gradually decreased. The above conclusion was consistent with the laws of substrate degradation and enrichment of functional microorganisms.

Modified glasgow prognostic score predicting high conversion ratio in opioid switching from oral oxycodone to transdermal fentanyl in patients with cancer pain.

The aim of this study was to identify predictive factors for higher conversion ratio in opioid switching from oral oxycodone to transdermal fentanyl (TDF) in patients with cancer pain. The participants of this study were 156 hospitalized cancer patients who underwent opioid switching from oral oxycodone to TDF at the Affiliated Hospital of Binzhou Medical University between January 1st, 2010 and March 31st, 2014. Patient characteristics, modified Glasgow Prognostic Score (mGPS), daily oxycodone dose, and reasons for opioid switching were retrospectively collected. The effect of variables on the conversion ratio was analyzed by multiple regression analysis to identify the predictive factors for higher conversion ratio in opioid switching from oral oxycodone to TDF. The results showed that the mGPS (odds ratio [OR], 2.358; 95% CI 1.379-4.031; P = 0.002), the reason for opioid switching (OR, 0.497; 95% CI, 0.298-0.828; P = 0.007) and equivalent oral morphine dose (OR, 1.700; 95% CI, 1.008-2.867; P = 0.046) were found to be significant predictors requiring higher conversion ratio in opioid switching. This study indicates that higher mGPS, poor pain control before switching and higher equivalent oral morphine dose are significant predictors of a need for higher conversion ratio in opioid switching from oral oxycodone to TDF. These results could contribute to the establishment of evidence-based medicine in cancer pain relief.

Poloxamer-188 attenuates TBI-induced blood-brain barrier damage leading to decreased brain edema and reduced cellular death.

Plasmalemma permeability plays an important role in the secondary neuronal death induced by traumatic brain injury (TBI). Previous works showed that Poloxamer 188 (P188) could restore the intactness of the plasma membrane and play a cytoprotective action. However, the roles of P188 in blood-brain barrier (BBB) integrity and TBI-induced neural cell death are still not clear. In this study, mice were induced TBI by controlled cortical impact (CCI), and cerebral water content was measured to explore the profile of brain edema after CCI. Further, the regimen of P188 in mouse CCI models was optimized. The neurological test and BBB integrity assessment were performed, and the numbers of TBI-induced neural cell death were counted by propidium iodide (PI) labeling. The expression of apoptotic pathway associated proteins (Bax, cyt-c, caspase-8, caspase-9, caspase-3, P53) and aquaporin-4 (AQP4) was assessed by RT-PCR or immunoblotting. The data showed that the brain edema peaked at 24 h after TBI in untreated animals. Tail intravenous injection of P188 (4 mg/ml, 100 µl) 30 min before TBI or within 30 min after TBI could attenuate TBI-induced brain edema. P188 pre-treatment restored BBB integrity, suppressed TBI-induced neural cell death, and improved neurological function. TBI induced an up-regulation of Bax, cyt-c, caspase-8, caspase-9, caspase-3, and the expression of p53 was down-regulated by P188 pre-treatment. AQP4 mainly located on endothelial cells and astrocytes, and its expression was also regulated by P188 pretreatment. All these results revealed that P188 attenuates TBI-induced brain edema by resealing BBB and regulating AQP4 expression, and suppressed apoptosis through extrinsic or intrinsic pathway. Plasmalemma permeability may be a potential target for TBI treatment.

Necrostatin-1 suppresses autophagy and apoptosis in mice traumatic brain injury model.

Traumatic brain injury (TBI) results in neuronal apoptosis, autophagic cell death and necroptosis. Necroptosis is a newly discovered caspases-independent programmed necrosis pathway which can be triggered by activation of death receptor. Previous works identified that necrostatin-1 (NEC-1), a specific necroptosis inhibitor, could reduce tissue damage and functional impairment through inhibiting of necroptosis process following TBI. However, the role of NEC-1 on apoptosis and autophagy after TBI is still not very clear. In this study, the amount of TBI-induced neural cell deaths were counted by PI labeling method as previously described. The expression of autophagic pathway associated proteins (Beclin-1, LC3-II, and P62) and apoptotic pathway associated proteins (Bcl-2 and caspase-3) were also respectively assessed by immunoblotting. The data showed that mice pretreated with NEC-1 reduced the amount of PI-positive cells from 12 to 48 h after TBI. Immunoblotting results showed that NEC-1 suppressed TBI-induced Beclin-1 and LC3-II activation which maintained p62 at high level. NEC-1 pretreatment also reversed TBI-induced Bcl-2 expression and caspase-3 activation, as well as the ratio of Beclin-1/Bcl-2. Both 3-MA and NEC-1 suppressed TBI-induced caspase-3 activation and LC3-II formation, Z-VAD only inhibited caspase-3 activation but increased LC3-II expression at 24 h post-TBI. All these results revealed that multiple cell death pathways participated in the development of TBI, and NEC-1 inhibited apoptosis and autophagy simultaneously. These coactions may further explain how can NEC-1 reduce TBI-induced tissue damage and functional deficits and reflect the interrelationship among necrosis, apoptosis and autophagy.

[Role of extracellular-signal regulated kinase cascade on cerebral ischemia and ischemic preconditioning in hippocampal neuron in gerbils].

AIM: To investigate the role of extracellular-signal regulated kinase (ERK) cascade on cerebral ischemia and ischemic preconditioning in hippocampal neuron. METHODS: Male gerbils were randomly divided into sham group (SH), ischemia/reperfusion group (I/ R), ischemia preconditioning group (IP), specific antagonist of ERK-PD98059 (PD), solvent control groups (VE group), PD98059 combined with IP group (PIP). Forebrain ischemia was induced by occlusion of bilateral common carotid arteries and confirmed by isoelectricity of EEG. Observations were carried out in each group 15 min, 2 h, 4 h, 6 h, 1 d, 3 d, 5 d and 7 d after ischemia. Open field test was used to examine the spontaneous motor activity, the survival and apoptotic neurons, Fos and NF-kappaB masculine neurons in hippocampal CA1 region were counted, the expression of HSP70 in hippocampal CA1 region and p-ERK in hippocampal CA3/DG regions were detected by SABC immunocytochemical technique. RESULTS: The spontaneous motor activity, the number of apoptotic neurons and NF-kappaB masculine neurons at 1 d, 3 d, 5 d, 7 d in CA1 region were much less in IP group than in I/R group (P < 0.01). The number of Fos masculine neurons at 15 min, 2 h, 4 h, 6 h, 1 d in CA1 region were significant more in IP group than in I/R group (P < 0.01). The expressions of p-ERK and HSP70 were significantly higher in IP group than in I/R group. The number of Fos masculine neurons at each point were more and apoptotic neurons at 1 d, 3 d were less in PD group than in I/R group. Results of observation in PIP group were within IP group and I/R group. CONCLUSION: Activation of ERK in CA3/DG regions were related to ischemic tolerance. Induction of the expression of Fos and HSP70, decreasing of the product of NF-kB which might be one of the molecule mechanisms playing an important role in neural protection of ischemic preconditioning.

[The relationship between the effects of curcumin on cerebral ischemia/reperfusion injury and immediately genic expressions of fos, Jun and NF-kappaB in hippocampal CA1 area and its significance in gerbils].

AIM: To explore the relationship between the effects of curcumin on cerebral ischemic/reperfusion injury and immediately genic expressions of Fos, Jun and NF-kappaB in hippocampal CA1 area. METHODS: Gerbils were randomly divided into sham group (SH), ischemia/reperfusion group (I/R), curcumin group (CU) and solvent control group (SC). Forebrain ischemia was induced by occlusion of bilateral common carotid arteries. Observations were carried out in each group 15 min, 1 h, 2 h, 6 h, 1 d, 3 d, 5 d and 7 d after ischemia: open field test was used to examine the behavioral change, the apoptosis neurons in hippocampal CA1 region was counted, the expression of Fos, Jun and NF-kappaB in hippocampal CA1 was detected by SABC immunocytochemical technique. RESULTS: The behavioral mark and the number of apoptosis neurons in hippocampal (CA1 region was much less in CU group than in I/R group (P < 0.01) The expression of Fos was more and the expression of Jun and NF-kappaB was less in CA1 area in CU group than in I/R group (P < 0.01). CONCLUSION: Curcumin can significantly protect neurons against cerebral ischemia, increasing the expression Fos and decreasing the expression of Jun and NF-kappaB may be the protective mechanisms.

A new peptide with membrane-permeable function derived from human circadian proteins.

Basic peptides such as human immunodeficiency virus type 1 (HIV-1) Tat-(48-60) and Drosophila Antennapedia-(43-58) have been reported to have a membrane permeability and a carrier function for intracellular protein delivery. Based on the fluorescence microscopic observations of the vascular endothelial cells (ECV-304) and the primary cultured neuroglial cells, we found that human Clock protein DNA-binding peptide [residue 35-47, hClock-(35-47)] had a translocation activity very similar to Tat-(48-60). The cellular uptake of hClock-(35-47) increases with the increase of incubation time and concentration. The internalization effect at 4 degrees was same as that at 37 degrees C. Internalization of hClock-(35-47) was saturable and could be inhibited by the excess of the other MPPs. Moreover, the uptake of these peptides were significantly inhibited in the presence of heparan sulfate. These results strongly suggested that the hClock-(35-47) shared a common or very similar internalization pathway with other MPPs. Furthermore, we injected rat through the common carotid artery with hClock-(35-47)-FITC peptide, and cryostat sections of the brain were prepared and observed using a fluorescence microscope. Result showed that the peptide had the ability to translocate through the blood-brain barrier. It is promising to provide a new safe carrier for the intracellular and encephalic treatment.