0.05) in tail flick test.MPE% in group MK was always higher than group M and descended more slowly than group M,especially from the d4 to d8(P0.05). Conclusion Ketamine could block the development of morphine tolerance partly due to its inhibition effect on pCREB protein.

Objective To investigate the expression of Toll-like receptor 4 (TLR4) and myeloid derived suppressor cells(MDSC) in bone marrow cells in children with acute myeloid leukemia (AML),and to detect its relationship with the clinical features,the effect of chemotherapy and prognosis.Methods Twenty-nine cases of children with AML were collected from June 2013 to March 2014 in People's Hospital of Wuhan University,in which 11 cases of low-risk group,10 cases of middle-risk group,8 cases of high-risk group;and 17 cases of non blood disease was as the control group.The expressions of TLR4 and MDSC were detected by using reverse transcription-polymerase chain reaction (RT-PCR),Western blot methods,immunohistochemical staining,and flow cytometry,respectively,in the bone marrow cells of 29 children with AML.Results The mRNA and protein expression of TLR4 in the initial treatment group was higher than those in the complete remission group(t =3.092,3.393,all P ＜ 0.05).The mRNA and protein expression of TLR4 in the relapse group was higher than those in the complete remission group(t =4.013,4.279,all P ＜ 0.05).The positive expression rates of MDSC in the above 3 groups were (29.77 ± 1.39) ％,(5.19 ± 0.65) ％,(38.62 ± 3.54) ％,respectively,compared with the control group [(1.32 ± 0.27) ％] and there was significant difference(all P ＜0.05).The positive expression rates of TLR4 and MDSC in the initial treatment group,relapse group and complete remission group were significantly higher than those in the control group,with significant differences (initial treatment group TLR4:t =3.559,P ＜ 0.05;MDSC:t =3.727,P ＜ 0.05;relapse group TLR4:t =4.043,P ＜ 0.05;MDSC:t =4.125,P ＜ 0.05;complete remission group TLR4:t =2.798,P ＜ 0.05;MDSC:t =3.469,P ＜ 0.05).Pearson rank correlation analysis showed that there was a positive correlation between the expression of TLR4 and MDSC (r =0.673,P ＜0.01).Conclusions The expressions of both TLR4 and MDSC play an important role in onset,progression,curative effect and prognosis in children with AML,and the two may play an importment role in synergistic effect.

Objective To investigate the molecular mechanism underlying lymphocyte functionassociated antigen-1 (LFA-1) / intercellular adhesion molecule-1 (ICAM-1) mediated anti-neoplastic effects of cytokine induced killer (CIK) cells. Methods Lymphocytes isolated from peripheral blood of children leukemia were induced with interferon-gamma (IFN-y), anti-CD3 monoclonal antibody (CD3McAb) and interleukin-2 (IL-2) and co-cultured with dendrite cells (DC) to generate DC-CIK cells. When treated with LFA-1 monoclonal antibody, cytotoxicity of DC-CIK cells against leukemia cell lines was measured by the MTT assay, while RT-PCR and Western blotting were used to determine mRNA and protein expressions of GATA-3 and T-bet in DC-CIK cells, respectively. IL-12, IFN-γ and tumor necrosis factor-α (TNF-α) levels released by DC-CIK cells were quantified by ELISA. Results Induced DC-CIK cells were regular, round and transparent with variable cell volume and cellular aggregation. When treated with mouse anti-human LFA-1 monoclonal antibody, the cytotoxicity decreased mostly towards B95 cells under administration of 20 μg/ml LFA-1 monoclonal antibody in comparison with the control group(t =10.138, P ＜0.05). It led to a highest elevation of GATA-3 mRNA and protein levels (t =16.386, P ＜ 0.05; t =22.652, P ＜ 0.05) and a most decrease of T-bet mRNA and protein levels (t =17.728, P ＜0.05; t =17.452, P ＜0.05) under 20 μg/ml LFA-1 monoclonal antibody in B95 cells group in comparison with the control group. The expression levels of IL-12,IFN-γ, and TNF-o in supernatant were the lowest under 20 μg/ml LFA-1 monoclonal antibody in B95 cells group in comparison with the control group (t =21.621, P ＜0.05; t =13.739, P ＜0.05; t =15.278, P ＜0.05).Conclusion GATA-3 and T-bet were implicated in the LFA-1/ICAM-1 mediated anti-neoplastic effects of DC-CIK cells via activation of the Th1 pathway, with high secretion of Th1 cytokines, such as IL-12, IFN-γ and TNF-α.

0 05). Conclusion Femal, splenomegaly and increase of the MPV width and PVP were the risk factors inducing PVT in liver cirrhosis patients, while liver function, BPC, PT, ect, may not be related to the formation of PVT.

Objective To clarify the relationship between ABO blood group and the development of duodenal ulcer(DU)in aspects of gastric acia and Hp infection.Methods The blood group of ABO was determined in 80 patients who were diagnosed as DU and received 24-hour gastric pH monitoring between 1995 and 2003.These results were compared with the expected frequency in the 1061 healthy controls in Beijing.The prevalence Helicobacter pylori (Hp) infection rate was determined by rapid urinase test,biopsy and 13C breath test.Results Blood type O was present in 56.3% of the patients with DU,which was significantly higher than the expected rate (28.7%) in healthy population (? 2=26.69,P0.05).Conclusion Blood group O doesn’t cause the disease by affecting Hp infection rate or stronger gastric acid secretion,it maybe another independent risk factor for DU.The onset age in blood group O is not different from that in other types.The mechanism needs to be explored.

Background Glial cell line derived neurotrophic factor (GDNF) is determined to have a neurotrophy effect and promoting effect to the growth of axon.GDNF has been applied in ophthalmology.Research showed that artemin,a new member of GDNF family,has a better function in protection of neuron,but seldom relevant document of distruibution of artemin in retina is found so far.Objective The aim of the present study is to investigate the distribution and expression of artemin in normal rat retinal neuron cells and retinal ganglion cells,and imitate diabetic environment to observe the expression of artemin at the condition of high glucose.Methods Retinal tissue was isolated from clean neonatal SD rats and cultured by expand culture method in DMEM/F12 containing 10% fetal bovine serum.40 mmol/L of glucose was added in medium in the seventh day after culture for 12 hours as experimental group.The expression and location of artemin in retina were tested by real-time PCR and cell immunofluorescence assay.Use of experimental animals followed the Management Regulation of experimental animals of Jiangsu Province.Results Cultured cells showed the typical cell body and processes in the seventh day.Cultured retinal ganglion cells (RGCs) presented the red fluorescence for Thy1.1 antibody,and multiple fluorescence label revealed that RGCs exhibited the green fluorescence for artemin antibody and red fluorescence for Thy1.1 antibody,indicating artemin protein was positively expressed in cultured RGCs.The numbers of positive cells for Thy1.1 antibody was (442±9)/high field in normal culture group and (263±7) /high field in 40mmol/L glucose culture group,showing a significant difference between them (P＜0.05).The expression of artemin mRNA in normal culture group and in 40 mmol/L glucose culture group,was showing a considerably difference between them(P＜0.05).Conclusion Artemin can be expressed in cultured retinal neuron cells and RGCs in rats.High glucose environment down-regulate the expression of artemin.This study proved a new idea for protecting RGCs against damage.

Objective To investigate the effects of nitric-oxide synthase inhibitor on matrix metalloproteinases (MMPs) expression in osteoarthritis (OA) cartilage by Luminex analysis,and to explore the mechanism of nitric-oxide synthase inhibitor on modification of the metabolism of OA cartilage.Methods Fifteen specimens of articular cartilage taken from the patients with OA were cultured and divided in two groups.The control group was those with no intervention.L-NIL group was co-cultured with NOS inhibitor L-NIL.After 72 h cultivation,the release of NO and the activity of NOS on OA cartilage were measured by Griess reaction and spectrophotometric methods.MMPs (MMP-1,MMP-2,MMP-3,MMP-9,MMP-13) expression was measured by Luminex analysis.Comparisons between groups were performed with paired sampies t test.Results After cultured for 72 h,spectrophotometric analysis showed high concentration of NO release[(216±47) μmol/L ] and high level of active NOS [(5.7±1.3)U/ml]in supernatants of the control,1 mmol/L concentration L-NIL could evidently reduce NO release [(55±20)μmol/L,P<0.01] and NOS activity [(1.7±0.7)U/ml,P<0.01 ].Luminex analysis demonstrated high MMP-1,MMP-2,MMP-3,MMP-9,MMP-13 expression in cartilages of the control group [respectively for (10.8±5.4)ng/ml,(9.2±3.3) ng/ml,[11.6±4.2 )ng/ml,(1.27±1.07)ng/ml,(3.6±1.3)ng/ml] and 1 mmol/L concentration L-NIL could evidently inhibit MMP-1,MMP-2,MMP-3,MMP-9,MMP-13 expression [respectively for (3.6±1.8)ng/ml,(2.3±1.2)ng/ml,(3.6±1.4)ng/ml,(0.65±0.21)ng/ml,(1.8+0.5)ng/ml,P<0.05 ].Conclusion Luminex analvsis has shown that NOS inhibitor can reduce NO release and NOS activity and modify metabolism of articular cartilage by inhibiting the over-expression of MMPs.

Objective To study the cause of death and mechanism after(TACE)in China during the past 14 years.Methods Related repots in Chinese Medical Current Content(CBM)and National Knowledge lnfrastruc ture(CNKI)from January 1994 to June 2008 were retrieved.The cause of death and mechainsm after TACE wer e analyzed.Results A total of 150 patients who died after TACE were reposed in China during the past 14 ye ar s.84%eases were caused by liver lunction failure,upper gastrointestinal bleeding and rupture of liver cancer. 78.7%cases died one month postoperation.Conclusion Liver function failure.upper gastrointestinal bleeding and rupture of liver cancer are the main complications which Can cause death and the majority cases died early.

Objective To investigate the effects of miR-200c on proliferation and apoptosis of tongue squamous cell carcino-ma (TCCS)Tca8113 cells.Methods The mimics of miR-200c were transfected into Tca8113 cells using liposome.The Tca8113 cell proliferation was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)assay.The flow cytometry as-say was used to determine the cell cycle and the apoptosis rate of Tca8113 cell.The protein expression levels of Bcl-2 and Caspase-3 in Tca8113 cell was detected by Western-blot.Results The 20,40,80 nmol/L miR-200c mimics groups inhibited the growth of Tca8113 cells,the difference compared with the control group showing statistical significance(P <0.05).The greater the miR-200c mimics concentration and the longer duration of action,the more significant the inhibition effect(P <0.05).After 48h transfecting by miR-200c mimics,the Tca8113 cells were arrested in the G0/G1 phases of cell cycle,and the apoptosis rate of the miR-200c mim-ics groups was significantly increased,the difference compared with the control group showing statistical significance(P <0.05);Western blot verified that the expression amount of Bcl-2 protein in the 20,40,80 nmol/L miR-200c groups was significantly lower than that in the control group,while the expression amount of Caspase-3 protein was significantly higher than that in the control group(P <0.05).Conclusion The overexpression of miR-200c might inhibit the proliferation of Tca8113 cell and induces their ap-optosis.

Objective To demonstrate the mechanisms underlying the efficacy of telmisartan in diabetic nePhroPathy, and discuss the role of PPARγin this Process. Methods The diabetic nePhroPathy rat models were established and randomly assigned to control grouP, telmisartan grouP ( 5 mg · kg-1 · d-1 ) and combination of telmisartan and PPARγ inhibitor grouP (telmisartan:5 mg·kg-1·d-1;GW9662:0. 5 mg·kg-1·d-1). After 12 weeks of treatment,the biochemical indexes of blood and urine,kidney weight,renal Pathology in each grouP of diabetic nePhroPathy rats were measured and comPared. The leVels of IL_1,IL_6 and TNF_α in blood of each grouP were detected by ELISA and comPared. The leVels of HGF and actiVated NF_κB (P65) in renal tissue of each grouP were detected by Western blotting and comPared. Results In diabetic nePhroPathy rats, telmisartan lowered the leVels of serum glucose, serum creatinine, urinary Protein and kidney weight, decreased the glomerular Volume,mesangial Proliferation and inflammatory cell infiltration,reduced blood leVels of IL_1,IL_6,and TNF_α,and decreased leVel of actiVated NF_κB (P65) in renal tissue. The leVel of HGF in renal tissue was eleVated by telmisartan. NeVertheless,these changes were Partly reVersed by PPARγ inhibitor GW9662. Conclusion PPARγ Presents an imPortant role in treatment of diabetic nePhroPathy by telmisartan.