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.

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.

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 To investigate the feasibility and applicability of free airbags,free fixed totally extraperitoneal laparoscopic inguinal hernia repair (TEP) treatment of inguinal hernia.Methods In Baotou Medical College,Department of General Surgery,the First Affiliated Hospital,the 40 cases of patients confirmed with inguinal hernia surgery were choosed from Jan.2010 to Dec.2012 as the observed object,depending on the surgical methods.Divided the cases into the TEP group and the open surgery group to observe statistics including postoperative pain,extent of gastrointestinal function recovery time,the time of bedside movement,the number of days of hospitalization,the incidence of postoperative complications,hospital fees indicators.Results The operation time of TEP group was about 20 minutes longer than the open group and the difference was statistically significant (P ＜ 0.05).Compared with the open group,TEP group' s postoperative pain was milder,postoperative feeding time,the time of bedside movement and postoperative exhaust time were earlier,hospital stays were shoter,incidence of postoperative complications was lower,the differences were statistically significant (P ＜ 0.05).The difference was not statistically significant in hospital costs(P ＞ 0.05).Conclusion TEP inguinal hernia greater advantages compared with traditional open surgery,may be appropriate to carry out in my area.

Schizophrenia, described as the worst disease affecting mankind, is a severe and disabling mental disorder. Schizophrenia is characterized by complicated symptoms and still lacks a diagnostic neuropathology, so developing schizophrenia animal models which have quantifiable measures tested in a similar fashion in both humans and animals will play a key role in new therapeutic approaches. According to the symptoms of cognitive impairment and emotional disorder, the N-methyl-d-aspartate (NMDA)-receptor antagonist MK-801 was applied to induce schizophrenia-like behavior in mice. Locomotor activity and prepulse inhibition (PPI) were selected as indices and the effect of clozapine was also investigated in this model. The results showed that compared with the normal group, MK-801-treated mice exhibited significantly increased locomotor activity and impaired PPI, and pre-exposure to clozapine could ameliorate the abnormality and make it back to normal level. These findings suggest that the model we established could be a useful tool for antipsychotic drug screening.

Aim To investigate the changes in the expression of protein kinase C gamma (PKC?), phosphorylated cAMP response element binding protein(pCREB)and immediate-early gene(c-fos and c-jun) in the spinal cord in formalin-induced inflammatory pain and study the effect of agmatine on the changes of PKC? activation, phosphorylation of CREB and expression of c-fos and c-jun.Methods Rats were decapitated at 10, 20 min or 2 h after intraplantar injection of 50 ?l 5% formalin and L_4, 5 spinal cords were dissected. Immunohistochemistry and western blotting analyses were used to observe the expression of PKC?, pCREB, c-fos and c-jun in the spinal dorsal horn and the effect of agmatine on the changes of their expression. Results Unilateral peripheral inflammation induced PKC? activation and CREB phosphorylation bilaterally while c-fos and c-jun expression ipsilaterally in rat spinal cord. PKC activity increased in membrane fractions with unchanged levels in the cytosolic fractions. Pretreatment intraperitoneally with 160 mg?kg-1 agmatine 15 min before inflammation significantly inhibited the activation of PKC? in the membrane fraction, suppressed the phosphorylation of CREB and the expression of c-fos and c-jun. Conclusion The mechanism of the analgesic effect of agmatine may be associated with inhibiting PKC? activation in the plasma membrane, CREB phosphorylation, c-fos and c-jun up-regulation which play roles in the hyperalgesia with peripheral inflammation.

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.

Objective To investigate the relationship between serum uric acid and traditional risk factors of coronary artery disease. Basic information was obtained by using questionnaire and measured for blood pressure , height ,weight. Venous blood was collected for detecting fasting plasma glucose ,serum lipid and uric acid. All data were analyzed with SPSS 20.0. Correlation analysis was used to assess the association of serum uric acid and traditional risk factors of coronary artery disease in patients with coronary artery disease. Results The mean value of serum uric acid in coronary artery disease is 6.3 mg/dL. The level of serum uric acid was significantly higher in male than that in female. Serum uric acid and body mass index(r = 0.137,P < 0.001),systolic blood pressure (r = 0.053,P = 0.025)and triglycerides(r = 0.188,P < 0.001)had a positive correlation. Serum uric acid was negatively correlated with high density lipoprotein cholesterol(r =-0.146,P < 0.001)and estimated glomerular filtration rate(r=-0.340,P<0.001). Serum uric acid was higher in patients with men,smoking,drinking,and chronic CAD. The serum uric acid level was lower in patients with exercise time more than 30 minutes than no movement or movement time less than 30 minutes. Conclusion Elevated serum uric acid were associated with body mass index,systolic blood pressure,triglyceride,high density lipoprotein cholesterol,estimated glomerular filtration rate,male,smoking,and chronic CAD.

Objective To evaluate T cell and CD4+ CD28-T in the development and progression of abdominal aortic aneurysms (AAAs).Methods Fifty Wistar rats were randomly divided into 5 groups (n =10 each).An AAA animal model is established by enhancing perfusing elastase to the infrarenal abdominal aorta of the rats.The levels of T cell,B cell and macrophage cell of abdominal aorta of the rats on days 3,7,14 and 28,were detected by enzyme linked immunosorbent assay(ELISA).CD4+ CD28-T cell of the peripheral blood were measured by flow cytometry.Result There was significant T-lymphocyte infiltration both in middle and outer membrane of the artery of the rats on day 7 after surgery.T-lymphocyte,B-lymphocyte and macrophage cell infiltration were on the peak in middle and outer membrane of the artery on day 14 after surgery.The ratio of CD4+ CD28-T cell in rat peripheral blood reached peak on day 7(P ＜0.05).Conclusions T cell and CD4+ CD28-T cell expression increased in peripheral blood and abdominal aorta in AAA rat model,suggesting a potential role of T cell and CD4+ CD28-T cells in the pathogenesis of AAAs,especially during the early development of AAAs.

Objective To observe the correlation of histologicalactivity(HAI) of chronic hepatitis B (CHB) with CCL20 expression, and to investigate the impact of CCL20 expression in CHB infection. Methods On the basis of established competitive quantitative RT-PCR with an internal standard, the expression of the CCL20 in the hepatocytes in different infected patterns of HBV infected cells and liver biopsies were quantified and at the same time its correlation to HAI were explored. Results In the cell levels, the expression quantity of CCL20 in control cells (HepG2), persistent HBV infected hepatocytes( HepG2. 2. 15) are (2. 65 ± 0. 02) pg/106 cells, ( 1.22± 0. 04) pg/106 cells, respectively. There were significantly differences between them ( t = 39. 66, P < 0. 01 ). The expression of CCL20 was enhanced in hepatocytes stimulated by PMA but their expression pauern was not changed. Moreover, CCL20 expression in liver biopsies with CHB was (3.54 ± 0. 65 ) pg/20 mg and CCL20 expression in control groups was ( 8. 74±0. 56) pg/20 mg. The expression of CCL20 between two groups was different (t =30. 09,P <0. 01 ) and correlation lied in between HAI and CCL20 expression in liver biopsies of CHB patients ( r = 0. 675, P =0. 023 ). Conclusion CCL20 expression was down-regulated and it was correlated to HAI of liver biopsies in CHB patients.