WS0701: a novel sedative-hypnotic agent acting on the adenosine system.

To characterize the sedative and hypnotic profile of the novel adenosine derivative ((3S,4R,5R)-3,4-dihydroxy-5-(6-((4-hydroxy-3-methoxybenzyl)amino)-9H-purin-9-yl)tetrahydrofuran-2-yl) methyl diaconate (WS0701), we performed a variety of behavioural tests and investigated the influence of WS0701 on various sleep stages. In mice, WS0701 significantly increased the number of entries and time spent in open arms in the elevated plus maze test, indicating an anxiolytic effect. WS0701 decreased locomotor activity counts and head dips in the hole-board test and enhanced sodium pentobarbital-induced hypnosis. However, WS0701 did not induce the loss of the righting reflex or amnesic effects in behavioural models. In rats, WS0701 exerted a sedative effect and markedly prolonged the time spent in non-rapid-eye-movement sleep, especially slow-wave sleep, but reduced the time spent in rapid-eye-movement sleep (REMS). Pretreatment with the selective adenosine A2a receptor antagonist SCH58261 attenuated the sedative and hypnotic effects of WS0701. WS0701 did not protect mice against picrotoxin-induced seizures, but inhibited adenosine deaminase activity and increased adenosine levels in the frontal cortex and hypothalamus of mice. In conclusion, WS0701 shows anxiolytic, sedative as well as sleep stage alterative effects, which may be related to the adenosine system.

Protective effects of the novel adenosine derivative WS0701 in a mouse model of posttraumatic stress disorder.

AIM: To investigate the effects of the novel N6-substituted adenosine derivative {(2R,3S,4R,5R)-3,4-dihydroxy-5-[6-[(4-hydroxy-3-methoxybenzyl)amino]-9H-purin-9-yl]tetrahydrofuran-2-yl} methyl decanoate (WS0701) on stress-induced excessive fear, anxiety, and cognitive deficits in a mouse model of posttraumatic stress disorder (PTSD). METHODS: Male mice underwent a conditioned foot shock and single prolonged stress procedure to induce PTSD. Contextual/cued fear, elevated plus-maze, open field and novel object recognition tests were conduced to assess PTSD-like behaviors. From d 1, the mice were orally administered WS0701 (7.5, 15, or 30 mg·kg(-1)·d(-1)) or paroxetine (10 mg·kg(-1)·d(-1)) for two weeks. Apoptosis of hippocampal neurons was detected using flow cytometry and TUNEL staining, and expression of Bcl-2 and Bax in the hippocampus was measured with Western boltting and qPCR assays. RESULTS: WS0701 administration significantly alleviated fear, anxious behaviors and memory deficits in the mouse model of PTSD. Furthermore, WS0701 administration significantly reduced the stress-induced apoptosis of hippocampal neurons, and increased the Bcl-2/Bax ratio in the hippocampus. The positive control drug paroxetine exerted similar effects on PTSD-like behaviors and hippocampal neuron apoptosis in the mouse model of PTSD, which were comparable to those caused by the high dose of WS0701. CONCLUSION: WS0701 effectively mitigates stress-induced PTSD-like behaviors in mice, partly via inhibition of neuronal apoptosis in the hippocampus.

Role of beta1-adrenoceptor in increased lipolysis in cancer cachexia.

Increased production of hormone-sensitive lipase (HSL) protein has been demonstrated to be the major cause behind enhanced lipolysis in cancer cachexia. The mechanism governing this alteration is unknown and was presently investigated. This study was conducted to detect the expression of relevant receptors in the adipocytes of cancer cachexia patients, and to elucidate their implication in the increased lipolysis. Gene expressions of beta1-adrenoceptor (ADRB1), beta2-adrenoceptor (ADRB2), beta3-adrenoceptor (ADRB3), alpha2C-adrenoceptor (ADRA2C), natriuretic peptide receptor A (NPRA), insulin receptor (INSR), and HSL were determined in adipose tissues of 34 patients by real-time PCR. Protein levels of ADRB1 and HSL were determined by western blot analysis. beta1-Adrenoceptor (ADRB1) was also detected by immunofluorescence staining. mRNA expressions of both ADRB1 and HSL were approximately 50% elevated selectively in the cachexia group, whereas mRNA levels of the other receptors were unchanged. beta1-Adrenoceptor (ADRB1) protein expression was 1.5-fold increased in cachexia as compared with the cancer controls, and 3-fold increased as compared with nonmalignant controls, and was confirmed as a membrane protein in adipocytes by immunofluorescence. Hormone-sensitive lipase (HSL) protein expression was 2-2.5-fold increased selectively in cachectic patients. There was a positive correlation between the protein expressions of ADRB1 and HSL. As much as approximately 50% of the variations in HSL protein expression could be explained by variations in ADRB1 protein expression. There was a link between ADRB1 protein level and lipolytic rate. Increased ADRB1 expression may account for some of the functional changes of HSL in patients with cancer cachexia.

Flavonoids from Rosaroxburghii Tratt prevent reactive oxygen species-mediated DNA damage in thymus cells both combined with and without PARP-1 expression after exposure to radiation in vivo.

This study aimed to evaluate the role of FRT in ROS/DNA regulation with or without PARP-1 in radiation-injured thymus cells. The administration of FRT to PARP-1-/- (KO) mice demonstrated that FRT significantly increased the viability of thymus cells and decreased their rate of apoptosis through PARP-1. Radiation increased the levels of ROS, γ-H2AX and 53BP1, and induced DNA double strand breaks. Compared with wild type (WT) mice, levels of ROS, γ-H2AX and 53BP1 in KO mice were much less elevated. The FRT treatment groups also showed little reduction in these indicators in KO mice compared with WT mice. The results of the KO mice study indicated that FRT reduced ROS activation through inhibition of PARP-1. Furthermore, FRT reduced the concentrations of γ-H2AX by decreasing ROS activation. However, we found that FRT did not regulate 53BP1, a marker of DNA damage, because of its elimination of ROS. Levels of apoptosis-inducing factor (AIF), exhibited no significant difference after irradiation in KO mice. To summarize, ROS suppression by PARP-1 knockout in KO mice highlights potential therapeutic target either by PARP-1 inhibition combined with radiation or by treatment with a drug therapy alone. AIF-induced apoptosis could not be activated in KO mice.

Protective effect of Xingnaojia formulation on rats with brain and liver damage caused by chronic alcoholism.

The aim of this study was to observe the effect of a formulation of traditional Chinese medicine extracts known as Xingnaojia (XNJ) on the liver function, learning ability and memory of rats with chronic alcoholism and to verify the mechanism by which it protects the brain and liver. A rat model of chronic alcoholism was used in the study. The spatial learning ability and memory of the rats were tested. The rats were then sacrificed and their brains and hepatic tissues were isolated. The activity of superoxide dismutase (SOD) and levels of glutamate (Glu), N-methyl D-aspartate receptor subtype 2B (NR2B), cyclin-dependent kinase 5 (CDK5) and cannabinoid receptor 1 (CB1) in the hippocampus were analyzed. The ultrastructure of the hepatic tissue was observed by electron microscopy. In addition, the activities of alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) in serum were tested and the levels of low-density lipoprotein (LDL), high-density lipoprotein (HDL), triglycerides (TG) and total cholesterol (TCHOL) were analyzed. XNJ enhanced the learning and memory of rats with chronic alcoholism. Treatment with XNJ increased the activity of SOD, and decreased the expression levels of NR2B mRNA and NR2B, CB1 and CDK5 proteins in the brain tissues compared with those in the model rats. It also increased the activity of ALDH in the serum and liver, decreased the serum levels of LDL, TG and TCHOL and increased the serum level of HDL. These results indicate that XNJ exhibited a protective effect against brain and liver damage in rats with chronic alcoholism.

[Impact of leachate recirculation loadings on efficiency of landfill gas (LFG) generation].

Effects of leachate recirculation loading on the efficiency of aerogenesis or methanogenesis of municipal solid wastes (MSWs) was investigated in four simulated anaerobic bioreactors (R1-R4), which were filled with 30 tons of wet weight waste each and recirculated weekly with 1.6, 0.8 and 0.2 m3 leachate and 0.1 m3 pure water, respectively. The results indicated that R1, with the highest recirculation ratio of 5.3%, began to produce landfill gas (LFG) largely after 5 weeks of leachate recirculation, while the other columns took 7-13 more weeks of lag phase time of LFG production. And LFG generation rates had good relationships with pollution loadings, such as COD and VFA in the leachate. By the 50th week, the waste in R1 was more stabilized with the highest loading rate. The accumulative transfer ratios to gas phase of TOC and COD were 28.96% and 14.57%, respectively, which meant large mount of organic matter was carried out by the effluent of the early stage and thus the potential of LFG generation was reduced. Therefore, to enhance the efficiency of LFG generation, the regimes of leachate recirculation in bioreactor landfills should be adjusted timely according to the phases of waste stabilization.

[Pilot scale research on impacts of leachate recirculation on its ultimate treatment].

Two pilot scale simulated columns, with and without leachate recirculation, were erected to study impacts of leachate recirculation of traditional anaerobic bioreactor landfill on leachate ultimate treatment methods. The results indicate that recirculation can remove organic pollutants visibly, but it isn't effective to inbiodegradable components, nitrogen (N) and phosphorus (P) pollutants. After recircualted for 44 weeks, test column has a BODs removal ratio of 98.5%. BOD5/COD value of outflow is only 0.07. BOD5/TN and BOD5/TP are 0.13 and 11 respectively, which are much lower than the adequate value for anaerobic microorganisms. It's difficult to treat this kind of leachate by traditional biological methods. When a bioreactor landfill is being designed, leachate characteristics after recirculated should be well considered and adequate leachate treatment, landfill and recirculation schemes should be chosen to take full advantage of waste stack decontamination effects.

[Effects of leachate recirculation volume on its characteristic and landfill stabilization rate].

Impacts of recirculation volume on leachate characteristic and landfill stabilization rate were studied. Four simulated bioreactor landfill columns were operated weekly with different recirculation ratios, respectively 5.3%, 2.7%, 0.67% leachate and 0.33% water, in this comparative research. The results indicate that simulated reactor with 5.3% recirculation ratio has the most rapidly stabilization rate and release the most organic pollutant. The shortest methane generation delay was also observed in this column. While simulated reactor with 2.7% recirculation ratio formed the best microbe environment and kept the highest reactor temperature 35 degrees C. It also had the best impactive load capacity and treating efficiency to leachate, and removal of COD was 77% and BOD5 was 88% respectively. In actual projects, appropriate leachate recirculation volume should be chosen according to design purpose of landfill sites.