Objective： To investigate the effects of CysLT receptor agonist leukotriene D₄(LTD₄) and antagonists on activation of microglia BV2 cells. Methods： The expression of CysLT₁ and CysLT₂ protein was determined by Western blotting and immunostaining in microglia BV2 cells.BV2 cells were pretreated with or without CysLT₁ receptor selective antagonist montelukast，CysLT₂ receptor selective antagonist HAMI 3379，or CysLT₁/CysLT₂ receptor dual antagonist BAY u9773 for 30 min，then the cells were treated with LTD₄ for 24 h.Cell viability was detected by MTT reduction assay.Phagocytosis and mRNA expression of IL-6 were determined by fluorescent bead tracking and RT-PCR，respectively. Results： In BV2 cells，LTD₄ did not affect proliferation but significantly enhanced phagocytosis and increased IL-6 mRNA expression in a concentration-dependent manner.LTD₄ at 100 nmol/L induced a 1.4-fold increase of phagocytic index and a 2-fold up-regulation of IL-6 mRNA expression（P<0.01）.HAMI 3379 and BAY u9773 (100 nmol/L) further increased LTD4-induced phagocytosis; BAY u9773 and montelukast decreased LTD4-induced IL-6 mRNA expression，while HAMI 3379 had no effect on that. Conclusion： LTD₄ activates BV2 cells in vitro and enhances IL-6 mRNA expression mediated by CysLT₁ receptor，LTD₄ induces phagocytosis which might be negatively regulated by CysLT₂ receptor in BV2 cells.

Objective： To evaluate the effects of aminguanidine on methylglyoxal-mediated oxygen-glucose deprivation (OGD) injury in the cultured human brain microvascular endothelial cells (HBMEC). Methods： Cultured HBMEC cells were pretreated with methylglyoxal before oxygen-glucose deprivation injury.Cell vitality was determined by MTT method，cell mortality was assessed by LDH release method，cell apoptosis was examined by AnnexinV/PI formation method，and the advanced glycation end products (AGEs) were detected by Western-blot. Results： Methylglyoxal induced HBMEC injury in a dose-dependent manner.At 2 mmol/L of methylglyoxa，the cell viability was 56.1%; when methylglyoxa-pretreated cells exposed to oxygen-glucose deprivation, the cell inhibition rate was 90.0%.Aminguanidine (1 mmol/L) inhibited methylglyoxa and OGD induced LDH release and AnnexinV/PI formation.Furthermore，aminguanidine(1 mmol/L) also decreased advanced glycation end products (AGEs) formation induced by methylglyoxal and oxygen-glucose deprivation. Conclusion： Aminguanidine protected methylglyoxal mediated-oxygen-glucose deprivation injury in the cultured HBMEC，which may be associated with anti-glycation activity.

Objective： To investigate the effects of aquaporin-4 (AQP4) gene knockout on the behavior changes and cerebral morphology during aging in mice,and to compare that of young and aged mice between AQP4 knockout mice (AQP4^-/-) and wild type mice (AQP4^+/+). Methods： Fifty-eight CD-1 mice were divided into four groups:young (2-3 months old) AQP4^-/-，aged (17-19 months old) AQP4^-/-，young AQP4^+/+ and aged AQP4^+/+.The activity levels and exploring behavior of mice were tested in open field.The neurons were stained with toluidine blue and NeuN，the astrocytes and microglias were stained with GFAP and Iba-1，respectively.The morphological changes of neuron，astrocyte and microglia were then analyzed. Results： Compared with young mice，the total walking distance in open field of aged AQP4^+/+ mice and aged AQP4^-/- mice decreased 41.2% and 44.1%，respectively (P<0.05); while there was no difference in the ratio of distance and retention time in the central area of open field.The density of neuron in cortex of aged AQP4^+/+ mice and aged AQP4^-/- mice decreased 19.6% and 15.8%,respectively (P<0.05)，while there was no difference in the thickness of neuron cell body in hippocampus CA1 region.The density of astrocyte in hippocampus CA3 region of aged AQP4^+/+ mice and aged AQP4^-/- mice increased 57.7% and 64.3%，respectively (P<0.001)，while there was no difference in the area of astrocyte.The area of microglia in hippocampus CA3 region of aged AQP4^+/+ mice and aged AQP4-/- mice increased 46.9% and 52.0%，respectively (P<0.01)，while there was no difference in the density of microglia.Compared with AQP4^+/+ mice，the young and aged AQP4-/- mice showed smaller area of astrocyte in hippocampus CA3 region，reduced 18.0% in young mice and 23.6% in aged mice.There was no difference between AQP4^+/+ mice and AQP4^-/- mice for other observed indexes. Conclusions： AQP4 may be involved in change of astrocyte and astrocyte-related behaviors during aging.AQP4 gene knockout may have limited effects on the change of neuron，microglia and most neuronal behaviors in aging process.

Objective： To identify a HEK293 cell line containing stably-transfected H3R gene，and to screen the novel non-imidazole compounds with H3R antagonist activity. Methods： The expression of rat H₃ receptor in cell line was detected by RT-PCR and Western blot.An elevation of intercellular cAMP concentration induced by forskolin was measured as the index for screening compounds with H3R antagonist activity. Results： The H3R-transfected HEK-293 cells stably expressed high level of rat H3 receptor mRNA and protein.Forskolin significantly increased intercellular cAMP concentration in the H3R-transfected HEK-293 cells.H3R agonist (R)-α-methyhistamine inhibited the forskolin-induced production of intercellular cAMP.H3R antagonist thioperamide and newly synthesized non-imidazole compounds XHA23 and XHA25 blocked (R)-α- methyhistamine reversal of forskolin-induced cAMP formation in a concentration-dependent manner，and the IC50 values were 3.62 μmol/L，0.49 μmol/L，0.14 μmol/L，respectively. Conclusions： The H3R-transfected HEK293 cells stably express high level of rat H3 receptor，and can be used for screening compounds with H3R antagonist activity．The non-imidazole compounds XHA23 and XHA25 may have H3R antagonist activity.

Objective： To investigate the roles of protease-activated receptors (PARs) in thrombin-induced brain injury and neurogenesis in rats. Methods： Ninety male SD rats were randomly assigned to receive intra-hippocampus injection of NS，thrombin or specific agonists of 3 protease-activated receptors (PAR-1，PAR-3 and PAR-4)，respectively.At 1，3 and 7 d after injection，the area of the hippocampus was determined with HE staining，the density and morphology of astrocyte were detected with GFAP staining，degenerated neurons were detected with Flouro-Jade C staining，and the neurogenesis was examined with DCX staining. Results： Compared to NS injection，the area of the hippocampus significantly increased at 1-3 d and decreased at 7 d after the injection of thrombin and PAR-1 agonist(P<0.05).In addition，injection of thrombin and PAR-1 agonist significantly increased the density of astrocyte and Flouro-Jade C positive cells at 1-7 d after injection(P<0.05)，and significantly increased the density of DCX positive cells at 3-7 d after injection(P<0.05).The injection of PAR-3 agonist and PAR-4 agonist had no affect on the area of the hippocampus，the density of astrocyte，Flouro-Jade C positive cells and DCX positive cells. Conclusion： The activation of protease-activated receptor-1 may be related to the thrombin-induced brain injury and neurogenesis in rat hippocampus.

Objective： To investigate the protective effects of carnosine against experimental closed head injury (CHI) in mice. Methods： The CHI model was established by free-falling weight-drop.Carnosine (250 mg/kg or 500 mg/kg) was administered intraperitoneally 30 min before brain trauma，then q.d for 7 d; while normal saline was administrated for control group.The neurological defect was evaluated by neurological severity score (NSS) within 7 d; the survival rate and the histological alternations were observed. Results： Carnosine prevented the body weight loss of mice at dose of 500 mg/kg; significantly increased the survival rate，and reduced the neurological defect and histological damage at dose of 250 and 500 mg/kg. Conclusion： Carnosine can attenuate closed head injury in mice.

Objective： Quantitative EEG and event-related potential P300 were used to evaluate impairment of cerebral function in patient with partial epilepsy. Methods： W value was calculated (power of EEG δ and θ rhythm divided by power of α and β rhythm) for the extent of focal cortical dysfunction.The W values in left partial epilepsy group，right partial epilepsy group and control group during interictal period were compared.The latency，amplitude and reaction time of P300 potential change were observed in each group. Results： The W values in F₈，T₄ and T₆ regions in patients with right partial epilepsy were higher than those in patients with left partial epilepsy（P<0.05）.The W values in T₃，O₁ regions of patients with left partial epilepsy were higher than those in patients with right partial epilepsy（P<0.05）.Furthermore，the W value in T₆ regions of patients with a disease course longer than 5y was significantly higher than that of patients with a disease course 1-5y or less than 1y; the W value in O2 regions of patients with a disease course longer than 5y was significantly higher than that of patients with a disease course between 1-5y（P<0.05）.In patients with right or left partial epilepsy，the total abnormal rate of P300 was 54.76%，the latency，amplitude and reaction time were significantly different to the control group.The abnormal rate of P300 in left and right partial epilepsy groups were 77.78% and 37.50%，respectively，and the former is significantly higher than the latter.The amplitudes of P300 in CZ and PZ of left partial epilepsy were significantly lower than those of right partial epilepsy and control group（P<0.05）.The latency and reaction time of P300 in CZ and PZ of all partial epilepsy were significantly longer than those of control group（P<0.05），however，no difference was found between left and right partial epilepsy. Conclusion： In partial epilepsy the cortical dysfunction occurs ipsilaterally to the epileptogenic zone，and extent of cortical dysfunction is positively correlated with duration of disease course.Cerebral dysfunction in left partial epilepsy is more severe than that in right partial epilepsy.

Objective： To observe the effect of recombinant human nicotinamide phosphoribosyl-transferase (NAMPT) on physiological/biochemical indexes and brain structure in mice. Methods： Wild type human recombinant NAMPT (10，30 and 100 μg/kg) or H247A mutant NAMPT (with very weak enzymatic activity) were administrated by intravenous injection in mice once every 3 d for 32 d.The changes of body weight，blood pressure，heart rate，serum glucose，serum total cholesterol and triglyceride were determined，and the morphology of neuron，astrocyte and microglia in hippocampus were observed. Results： The injection of wild and mutated type NAMPT had no significant effect on body weight，blood pressure，heart rate，blood glucose，total cholesterol and triglyceride，and did not affect the morphology of neuron，astrocyte and microglia in hippocampus of mice. Conclusion： Elevation of plasma NAMPT may not induce metabolic and neuronal dysfunction in normal individual.

Objective： To isolate，culture and identify of human endometrial stromal stem cells. Methods： Endometrial tissue was obtained from 10 women undergoing hysterectomy.Purified stromal stem cell suspensions were then obtained by selecting cells with magnetic bead sorting and colony-forming.The surface markers of stromal cell were identified by flow cytometry.Proliferation of stromal stem cell was observed by MTT methods.The osteogenic potential was evaluated by alizarin red staining，the adipogenic potential by oil red O staining.Then the adipogenic and osteogenic specific markers of differentiated cells assayed by RT-PCR method.Expression of cell surface antigen OCT-4 was detected with immunocytochemical staining. Results： Endometrial stem cells were successfully isolated from human endometrial tissue.They were stably proliferated and subcultured in vitro.Most of the passage cells expressed mesenchymal stem cell markers CD90，CD105 but not hemopoietic markers CD34，CD45.The analysis of cell cycle indicated that the percentage of G2-M phase and S phase cells increased.The growth curves of the third passage presented in "S" shape.After cultured in differentiation medium，the cells differentiated toward adipoblasts and osteoblasts as verified by positive staining with Oil Red O and alizarin red staining.Under induction，cells expressed osteogenic and adipogenic marker genes.The immunocytochemical staining of OCT-4 was positive. Conclusion： Human endometrium contains endometrial stromal stem cells，which present characteristics of mesenchymal stem cells and may be used as seed cells for tissue engineering.

Objective： To investigate the seasonal variation of total flavonoid content of Farfujium japonicum and its response to stress. Methods： The total flavonoids of Farfujium japonicum were determined by spectrophotometry in different seasons and under various stressful factors. Results： The total flavonoid content in Farfujium japonicum leaves was the highest，followed by the petiole，and rhizomes (P<0.05).The total flavonoid content in the leaves in December was higher than that in other months; that in the petiole and rhizome fluctuated in different seasons (P<0.05).As the light intensity enhanced，the total flavonoids in Farfujium japonicum leaves，petioles，rhizomes showed a downward trend.With the increase of water stress，the total flavonoid content in Farfujium japonicum leaves gradually increased，that in petiole first increased and then decreased，while that in rhizomes decreased (P<0.05).With the salt stress，the total flavonoid content in leaves，petioles and rhizomes of Farfujium japonicum showed a decreasing trend (P<0.05).With the increasing of temperature，the total flavonoid content in the leaves showed a gradually increasing trend; that in petiole first decreased and then increased，while that in the rhizomes first increased and then decreased (P<0.05). Conclusion： The total flavonoids of Farfujium japonicum fluctuate with the change of seasons and that in different parts of the plant has different responses to ecological stressful factors.

Objective： To investigate the protective effect of dexmedetomidine (Dex) preconditioning against ischemia/reperfusion (I/R) injuries in isolated rat hearts and its relation to mitochondrial permeability transition pore (mPTP) and mitochondrial ATP-sensitive K+ channel (mitoK_ATP). Methods： The hearts of male SD rats were isolated to mount on the Langendorff apparatus and subjected to 30 min global ischemia followed by 120 min reperfusion.The isolated hearts were treated with Dex (10 nmol/L) before ischemia for 15 min.The left ventricular hemodynamic parameters,coronary flow (CF) and the lactate dehydrogenase (LDH) release in the coronary effluent at 5 min reperfusion were measured.The formazan content was assayed to determine the myocardial viability at the end of reperfusion. Results： Compared with normal controls,I/R markedly decreased the left ventricular developed pressure and CF during the whole reperfusion period and the formazan content; while the left ventricular end diastolic pressure and LDH release were significantly increased.Dex preconditioning markedly improved the myocardial viability and cardiac function (P<0.01),which were reversed by the treatment with both atractyloside (20 μmol/L before ischemian),an opener of mPTP,and 5-hydroxydecanoate (100 μmol/L at the beginning of reperfusion),an inhibitor of mitoKATP,for 20 min. Conclusion： Dex has protective effect against I/R injuries in isolated rat hearts,which may be related to inhibiting the opening of mPTP at the beginning of reperfusion and activating mitoK_ATP before ischemia.

Objective： To investigate the biological characteristics of phage SM1 for stenotrophomonas maltophilia (Sm) and its effect in animal infection model. Methods： Phage SM1 isolated from raw sewage of hospital was identified by the plaque method.The morphology of phage SM1 was observed by electronmicrographics with negative staining.The extraction and electrophoresis of phage SM1 DNA were performed.Optimal multiplicity of infection，resistant mutation rate，one step growth curve and the effectiveness in animal models of phage SM1 were determined. Results： One Sm specific phage of myoviridae double-stranded DNA was identified，and named SM1.Electrophoresis of DNA demonstrated that the size of phage SM1 genome was about 50 kb.The growth curve of phage SM1 showed that the durations of incubation and burst period were 15 min and 50 min，respectively; and the burst size was 187.The resistant mutation rate of phage SM1 was 6×10^-10.All mice treated with phage SM1 survived after 7 d of infection with stenotrophomonas maltophilia. Conclusions： The phage SM1 has a relatively broad host range，a shorter incubation period，an apparent burst size and a lower resistant mutation rate.The therapy of phage SM1 for Sm infection in mice is effective.

Objective： To assess the application of gray-scale contrast-enhanced ultrasound (CEUS) and contrast-enhanced spiral computed tomography (CECT) in detection of residual tumor after high intensity focused ultrasound (HIFU) ablation with microbubbles on rabbit hepatic VX2 tumors. Methods： Forty rabbits with hepatic VX2 tumors were randomly divided into three groups before ablation.Group Ⅰ (n＝10) served as sham ablation controls，rabbits in group Ⅱ (n＝15) and group Ⅲ (n＝15) were ablated using HIFU under the manipulation of computer.A bolus of 0.2 ml SonoVue solution was injected via ear marginal vein of rabbits in group Ⅲ before ablation.Tumors were examined with CEUS and CECT before and within 3h after HIFU ablation.Necropsy and histopathological assessment were performed immediately after the completion of images evaluation. Results： Before ablation，intense arterial feeding vessels was detected in the tumors (77.5%，31/40 vs 52.5%，21/40) or the periphery of the tumors (22.5%，9/40 vs 47.5%，19/40) by CEUS and CECT，respectively.The tumors were characterized by quick wash-in and wash-out (high and rapid peak of enhancement in the arterial phase，followed by a fast decrease in enhancement level).The dose parameters used to achieve therapeutic effect in group Ⅲ were significantly lower than those in group Ⅱ(P＜0.01).There were local residual viable tumor tissues due to incomplete ablation in 60.0% (9/15) of group Ⅱ and 13.3% (2/15) of group Ⅲ revealed by histopathology(P＜0.05).The concordance rate of CECT and CEUS with histopathology on residual tumor detection was 27.3% and 81.8% (P＜0.05)，respectively. Conclusions： The administration of microbubble agent enhances the efficacy of HIFU on rabbit hepatic VX2 tumors.CEUS is more sensitive than CECT in detection of residual viable rabbit VX2 tumor after HIFU.

Objective： To investigate the association of hepatitis C virus (HCV) genotype with glycolipids iron metabolism in Gansu Han population. Methods： The genotypes of HCV 1b type and 2a type were detected in Gansu Han HCV carriers.The Glu，Insulin，CHOL，TG，UIBC，TRF，TIBC，SF，Serum Iron，AST，ALT，TBil，IBil，DBil，ALP，GGT were measured and compared between patients with different HCV genotypes. Results： There were 84 cases with HCV1b type and 136 cases with 2a type.There were significant differences in TG，ALT，TRF，TIBC between 1b type and 2a type genotype HCV carriers. Conclusion： The 2a type HCV carriers may be more inclined to develop hyperlipidemia and liver damage，and 1b type HCV carriers are likely to have iron metabolism defect.

Objective： To evaluate the methods for collection of peripheral blood stem cells (PBSC) in children. Methods： Peripheral blood stem cells were collected from 20 child patients and 11 donors.The patients treated with chemotherapy，received G-CSF or GM-CSF and the donors received G-CSF for mobilization.When the peripheral blood (PB) leukocyte count reached to 5×10⁹/L,the hematopoietic stem cells were collected with CS-3000 Plus，COM TEC or COBE Spectra blood cell separators from patients and donors.For children whose weight＜20 kg，HCT<24% or TBV≤1 100-1 650 ml，blood cell separators were pre-injected with the same type RBCs irradiated by 25 Gy of gamma-ray and with low flow rate (10-30 ml/min).The number of CD34⁺ cell was detected by flow cytometry.The relationship of number of CD34⁺ cell with mononuclear cell (MNC) and processed blood volume was analyzed. Results： Successful collection of the PBSCs with the CS- 3000 Plus (n=10)，the COM TEC (n=3) and the COBE Spectra (n=18) was achieved in all the 31 cases with 1-5 aphereses used.Number of CD34⁺ cells was（7.9±2.9）×10⁶/kg and that of MNCs was（7.4±3.1）×10⁸/kg.The total CD34⁺ cell count was correlated with MNCs before aphaeresis and processed blood volume. Conclusion： For collection of high quality PBSCs，the appropriate methods should be chosen according to the body weight，TBV，mobilization of child patients/donors.

G-protein-coupled receptor 17 (GPR17)，an originally orphan receptor，was identified as a new uracil nucleotides/cysteinyl leukotriene receptor.However，whether GPR17 is really classified as a leukotriene receptor is a matter deserving further investigation.GPR17 is involved in many physiological and pathological processes including brain injury，spinal cord injury，and oligodendrocyte differentiation.GPR17 may become a new therapeutic target in these diseases.In this article，the research progress on the pharmacology and pathophysiological roles of GPR17 is reviewed.

Colorectal cancer (CRC)，one of the most common malignant tumors，is caused both by environmental and genetic factors.Genetic factor plays an important role in the pathogenesis of CRC.Genome-wide association study (GWAS) is a new tool of genetic research.A series of susceptibility genes and loci of the complex diseases has been identified with GWAS strategy.In this article，the research progress on GWAS of CRC is reviewed，and the advantages and limitations of GWAS study as well as the prospective of its application are discussed.