Opioid peptides and their receptors play an important r ole in the pathophysiologic process of stroke. The concentration of endogenous o pioid peptides elevates, while the density of opioid receptors decreases in CNS during cerebral ischemia; on the other hand, opioids influence the development o f stroke through regulating the contraction of cerebral vessel, immuno-endocrin e system and neuroprotection against injury. Thus, drugs that regulate the opioi d system may be a new therapeutic approach for prevention and treatment of cereb ral ischemia.

As an important messenger molecule, NO exists in both peripheral and central nervous system (CNS). Recently, NO was proved to play a critical role in CNS, especially in synaptic plasticity. As a kind of retrograde messenger, it plays some roles in the induction and maintenance of LTP through activation of downstream pathway or molecules, such as NO-cGMP-PKG, ADPRT and C-FOS.

Aims To establish several behavioral paradigms to characterize the psychotropic effects of hallucinogens which ze-brafish was utilized as a model animal, and then to investigate the effects of potent hallucinogenic drugs on these models. Methods With the video record and track system, the behavior was recorded and quantified automatically. In the experiments, the bottom dwelling test, social behavior and mirror test were performed to test the hallucinogenic effects of drugs. Metham-phetamine (METH, 2 mg·L-1) and ketamine (20 mg·L-1) were selected as experimental challenges. The 30 min pre-treat-ment time was chosen based on our prior experience in zebrafish models. Results Compared to the normal group, in dwelling test, acute exposure of zebrafish to METH and ketamine de-creased transitions significantly, and in mirror reflection test, the drug-treated fish changed the preference for mirror zone, and ex-hibited aggressive for their mirror images. The pretreatment of METH and ketamine significantly reduced the contact durations, and the ketamine inhibited the contact frequency each other, the results indicated that the social interaction of zebrafish was im-paired. Conclusion The results confirm high sensitivity of ze-brafish models to hallucinogenic compounds with complex behav-ioral and physiological effects.

Psychostimulant is a type of psychoactive substances that stimulate central and peripheral nervous system as well as induce drug dependence.A series of studies indicate that monoamine especially dopamine plays an important role in behavior and drug dependence,moreover,dopamine transporter(DAT)controls homeostasis of dopamine in neuron and transmission of dopamine pathways.Thus DAT might play an important role in the reward and behavioral stimulation of psychostimulant.

OBJECTIVE Toestablisharecombinantcelllinethatcanstablyexpressratpurinergic P2receptorP2X4(rP2X4R).METHODS ToconstructgreenfluorescentproteinandrP2X4recombinant plasmid (pEGFP-N1-rP2X4),lipofectamine was used to transfect pEGFP-N1-rP2X4 into human embry-onic kidney (HEK293)cells that were screened with G41 8 (1 g·L-1 ).The quantitative expression of rP2X4 receptor was verified by qRT-PCR and Western blotting analysis.Whole-cell patch clamp record-ing was used to investigate the function of the stably expressed rP2X4 receptor. RESULTS The sequence of plasmid pEGFP-N1-rP2X4 was verified by PubMed Blastn comparison.qRT-PCR and Western blotting analysis demonstrated that the expression of rP2X4 receptor in HEK293-pEGFP-N1-rP2X4 cell lines remained stable after 25 generations (P1 ,P3,P5,P10,P15,P20 and P25).Whole-cell patch clamp recording experiments showed that the rP2X4 receptor agonist,purine-5′-triphosphate (ATP,3.0 μmol·L-1 ),could activate rP2X4 receptors in HEK293-pEGFP-N1-rP2X4 cell lines.Specific activating current could be blocked by non-selective rP2X4 receptor antagonist TNP-ATP (30.0μmol·L-1).CONCLUSION rP2X4receptorisstablyexpressedinHEK293-pEGFP-N1-rP2X4cell line and maintains stable expression and function within 25 continuous generations.The establish ment of HEK293-pEGFP-N1-rP2X4 cell line can contribute to further investigations of the roles of rP2X4 receptors in neuropathic pain.

25% signified analgesic effect. Part II (the effect of agmatine on analgesic effect of morphine). Fifty-six animals with neuropathic pain were randomly divided into 4 groups ( n = 14 each) . Each group received NS or agmatine 20, 40 or 60 mg? kg-1 . 30 min later each group received either NS or morphine 1 mg?kg-1(n=7 each). PWT was measured and PMAP was calculated Part III (the effects of agmatine on the development of morphine tolerance). Morphine tolerance was incuced by morphine 10 mg?kg-1, 3 times a day for 5 days. Thirty-five animals with neuropathic pain were randomly divided into 5 groups ( n = 7 each): group A received NS: group B received NS + morphine; grouop C, D, E received agmatine 5, 10, 20 mg ? kg-1 + morphine 3 times a day for 5 days. PWT was measured on the 5th day and PMAP was calculated. Results PMAP of agmatine (40, 60 mg?kg-1 i.p.) and morphine (1 mg?kg-1 s.c.) were lower than 40% . Agmatine 60 mg?kg-1 increased the PMAP of morphine 1 mg?kg-1 to 84% compared with 34% for morphine alone (P

AIMTo further elucidate the role of agmatine on the pharmacological effects of opioids. METHODSThe effect of L-arginine and L-arginine decarboxylase(L-ADC) antibodies on pain threshold, morphine ntinociception and tolerance were investigated in mouse acetic acid writhing test, mouse radiant heat tail flick test and mouse hot plate test. RESULTSIn mouse acetic acid writhing test, intracerebroventricular injection of L-arginine dose-ependently inhibited the writhing of mice compared with saline control. L-arginine did not influence the tail flick latency itself in mouse radiant heat tail flick test, but enhanced antinociceptive effect of morphine in a dose-dependent manner. The possible maximal analgesia percentage of morphine 2.5 mg*kg-1 was increased from 23% to 71%. Furthermore, L-arginine inhibited acute tolerance induced by morphine 100 mg*kg-1in mouse radiant heat tail flick test. The effect of L-arginine as mentioned above could be antagonized by idazoxan (3 mg*kg-1, ip), which is a selective antagonist of imidazoline receptors. L-ADC specific antibodies inhibited morphine antinociception and promoted the development of tolerance to morphine in mouse radiant heat tail flick test and 55℃ hot plate test. CONCLUSIONL-Arginine and L-ADC play important roles in the formation of pain threshold, morphine antinociception and tolerance.

Cas9 is a RNA-guided double stranded DNA nuclease that participates in the CRISPR/Cas9 system. Wide-type Cas9 directly silences the expression of target gene by gene splicing. The engineered dCas9 protein with the mutation at D10A and H840A lacks the Cas9' s endonuclease function but keeps its DNA binding activity. dCas9 can activate special genes by fusing with transcription activator. Meanwhile,it can inhibit the gene transcription by directly binding to the target gene and stop gene transcrip?tion. Combination of light sensitive structures and CRISPR can produce light-inducible CRISPR/Cas9 system for control of gene expres?sion. This system is able to activate or inhibit gene expression via the use of controlling blue light(470 nm). In this review,we mainly discuss the development of the light inducible CRISPR/Cas9 system as well as its application in the control of gene expression.

OBJECTIVE To explore the antidepressant effect and the underlying mechanisms of schisandrin (SCH), a component of the fruits of Schizandra chinesis. METHODS The forced swimming test (FST) and tail suspension test (TST) in mice were used to evaluate the antidepressant activity of SCH (5, 10, and 30 mg · kg-1) following single administration intragastrically, and the locomotor activity was investigated to exclude its neural excitatory effects. Effects of SCH on neural monoamine systems were studied in two pharmacological models, including reserpine induced monoamine depletion test and yohimbine toxicity potentiation test. RESULTS In behavioral despair models, SCH (30 mg·kg-1) signif?icantly decreased the immobility time in the TST and FST (P<0.05) compared with normal control group. Results of the locomotor activity experiment showed that SCH had no excitatory or inhibitory actions on the central nervous system. In the reserpine reversal experiment, SCH (30 mg · kg-1) antagonized thepalpebral ptosis and akinesia symptoms caused by reserpine(2.5 mg · kg-1) treatment (P<0.05) compared with model group, but had little effect on the drop of the anal temperature. Moreover, SCH did not increase the lethality caused by subcutaneous injection of yohimbine (30 mg · kg-1)at the threshold lethal dosage. CONCLUSION SCH exerts potential antidepressant-like effect in mice.

AIM: To investigate the effect of ecdysterone (EDS) on H9c2 cardiomyocytes after oxidative stress.METHODS:H9c2 cells were cultured in vitro and divided into control group, high dose (2 μmol/L) of EDS group, middle dose (1.5 μmol/L) of EDS group, low dose (1 μmol/L) of EDS group, and H2O2 group.H9c2 cardio-myocytes in H 2 O2 group and high , middle and low doses of EDS groups were exposed to H 2 O2 for 6 h to establish the model of oxidative stress.The viability of the H9c2 cells was detected by CCK-8 assay.The apoptosis of H9c2 cells was analyzed by flow cytometry.The levels of lactate dehydogenase (LDH) and creatine kinase-MB (CK-MB) in the culture medium, and the levels of superoxide dismutase (SOD) and malondialdehyde (MDA) in the H9c2 cells were measured by colorime-try.The generation of reactive oxygen species (ROS) and the mitochondrial membrane potential were evaluated by flow cy-tometry and confocal laser scanning microscopy .The protein levels of Bax , Bcl-2 and cleaved caspase-3 in the H9c2 cells were determined by Western blot .RESULTS:Ecdysterone at the selected concentrations had no effect on the viability of H9c2 cells.Compared with control group, the levels of LDH, CK-MB, ROS and MDA, and the apoptotic rates of the H9c2 cells were significantly increased after treated with H 2 O2 , but were decreased by EDS treatment in a dose-dependent man-ner.The levels of SOD and mitochondrial membrane potential of the H 9c2 cells in H2 O2 group were reduced significantly compared with control group , but high, middle and low doses of EDS treatments up-regulated the levels of SOD and mito-chondrial membrane potential in H 2 O2-treated H9c2 cells.The protein levels of Bax and cleaved caspase-3 in the H9c2 cells in H2 O2 group showed significant elevation in comparison with control group , and the protein expression of Bcl-2 de-clined in H2 O2 group compared with control group , but high, middle and low doses of ecdysterone treatments down-regula-ted the protein levels of Bax , cleaved caspase-3 and up-regulated the expression of Bcl-2 in H2 O2-treated H9c2 cells. CONCLUSION:Ecdysterone attenuates the effect of H 2O2-induced oxidative stress on H9c2 cardiomyocytes.The mecha-nism may be involved in scavenging oxidative stress products , increasing antioxidant enzyme activity and improving mito-chondrial function .