Objective To determine the optimal dose range of naloxone to enhance the analgesic effect of morphine.Methods One half of a total of 84 adult male Sprague-Dawley rats were randomly assigned into seven groups(6 rats for each group).Rats in group NS received normal saline,and in group M received 6mg/kg of morphine.Different doses of naloxone(1?g/kg,100ng/kg,10ng/kg,1ng/kg and 0.1ng/kg)with 6mg/kg of morphine were given to the rats in group MN1,group MN2,group MN3,group MN4 and group MN5.Pain thresholds were determined at different time points before and after subcutaneous injection of normal saline or morphine or mixture of the drugs(morphine and naloxone).Another 42 rats were randomly assigned into seven groups similar to the above grouping,but the morphine doses for group M and groups MN were changed to 2mg/kg.Acute pain was prodused by an in cision on the hind paw.Then they were given subcutaneous injection of the drugs in different doses as categorized above.Cumulative pain scores were observed within an hour.Results Compared with group NS,the pain thresholds of all the other groups were significantly increased at the time points from 5 minutes to 120 minutes after subcutaneous injection(P0.05).Conclusions Low-dose of naloxone can enhance the analgesic effect of morphine,and the dose range 1ng/kg~100ng/kg may be acceptable.Dose of 1?g/kg naloxone may antagonize the analgesic effect of morphine,while dose of 0.1ng/kg naloxone,perhaps,is too low to show an effect.

0.5 mV and change in color of myocardium. Blood samples were taken from right atrium for determination of plasma SOD activity and plasma MDA level and from coronary sinus and artery for determination of blood lactate level before occlusion of LAD ( T0 ) , before reperfusion (T1),1,2,3,4,5,6 h after reperfusion (T2-7 ) . Myocardial lactate production was calculated from the difference between coronary sinus and arterial blood lactate concentrations. Results ( 1) In HTEA group HR, MAP and CVP decreased by 22% , 25% and 28% after epidural blockade, while in control group there was no significant change after epidural saline. (2) In HTEA group plasma SOD activity started increasing at T6 and blood MDA level decreased at T4 and T5, whereas in control group blood SOD activity started decreasing and blood MDA level started increasing at T3 . (3) Myocardium released no lactate before ischemia. Myocardial lactate release greatly increased during ischemia and started decreasing after reperfusion in both groups. But myocardial lactate production was significantly less in HTEA group than that in control group. (4) One animal died from ventricular fibrillation at the beginning of reperfusion in HTEA group while in control group four animals died. Conclusion HTEA can alleviate the myocardial ischemia-reperfusion injyry by blocking sympathetic nervous activity.

Objective To study the effects of whole-body irradiation(WBI)with different doses of X-rays on apoptosis in mouse splenocytes and peritoneal macrophages.Methods The apoptosis percentages of mouse splenocytes and peritoneal macrophages were detected with flow cytometry(FCM)at different time after the whole-body X-irradiation using the staining of Annexin-V and PI.Results As compared with the control,the percentage of apoptosis in mouse splenocytes began to increase gradually 24 h after WBI with 2 Gy X-rays(P

0.05).The apoptotic percentage was increased significantly after X-rays irradiation with the dose of 6 Gy(P

Objective: To investigate the ratios of peripheral blood CD4+CD25+,CD4+CD8+ regulative T cells of systemic lupus erythematosus(SLE) patients, and explore the association with disease active stage,nephropathy,serum anti-ds-DNA antibody,and both IgG and C3 levels. Methods: The percentage of CD4+CD25+T cells and CD4+CD8+T cells of peripheral blood from patients with systemic lupus erythematosus(SLE)(30 females and 7 males),30 rheumatism controls and 30 normal individuals were measured by flowcytometry. Results: Patients with active disease had statisitically lower levels of CD4+CD25+T cells than did normal controls(P

Objective To study the influence of sampling,handling,shipping and storage on low-molecular-weight serum proteome profiling.Methods Serum samples instantly separated and aliquoted,some stored at-80 ℃ for up to 6 months,others stored at 4 ℃ or room temperature(25 ℃) for 2 to 72 hrs.The variations of protein profiling under these conditions on WCX magnetic beads were studied.Profiling influenced by hemolysis,multi freeze-thaw cycles,storage conditions.Results Different handling procedures and storage conditions have different effects on serum profiling.Serum stored at-80 ℃ up to 6 months,stored at 4 ℃ or 25 ℃ for 2 h or one freeze-thaw cycle,had little effects on serum proteomic analysis.If serum diluted into 9 mol/L urea buffer at room temperature,the result is stable for 24 hours.Statistics analysis shows that 35.9% peaks have significant changes(P

Objectives To investigate the ratios of peripheral blood CD4+CD8+ and CD4+CD25+ regulative T cells, and explore the association with hepatic damnification and anti-AMA-M2 antibodies.Methods The percentage of CD4+CD8+T cells and CD4+CD25+T cells in peripheral blood from patients with primary biliary cirrhosis(PBC) (n=27)、26 patients with other hepatic desease、30 normal individuals were measured by flowcytometry.Results Patients with PBC had statistically higher levels of CD4+CD25+T cells than the patients with other hepatic disease (P

Objective To study the relationship between the expression of eostimulating factor B7-H4 in patients with primary biliary cirrhosis (PBC) and the pathogenesis of PBC. Methods The expression of B7-H4 mRNA on peripheral blood mononuclear cell (PBMC) of 65 patients with PBC was tested by real-time PCR. Serum levels of IL-2 were assayed by ELISA. CD4+, CD8+ T lymphocytes expression level and B7-H4 expression rate before and after activation were measured by three-color flow cytometry (FCM). Re-sults (1) Expression of B7-H4 mRNA and BT-H4 percentage in PBC group were significantly lower than that in none-PBC group and healthy controls(P<0.01);(2)After 72 h activation, the percentage of CD4+, CD8+, and CD4+ CD8+T lymphoeytes and serum levels of IL-2 decreased (P<0.05), and the percentage of CD4+, CD4+ CD8+T lymphocytes and serum levels of IL-2 were significantly higher than that of none-PBC group and healthy controls(P<0.01);(3)Levels of alanine aminotransferase(ALT), aspartate amin-otransferase(AST), alkaline phosphatase(ALP) and γ-glutamyl transpeptidase(GGT) of patients with posi-tive anti-mitochondrial antibody(AMA)-M2 rose. There was no significant difference of B7-H4 expressions on T cells between patients either with or without AMA-M2 antibody. Conclusion The costimulating factor B7-H4 can express on T lymphocytes which is activated by phytohemagglatinin(PHA) aad plays a negative role on T cells responses.

Objective To clone and express the human asialoglycoprotein receptor(ASGPR) H1 subunit, purify and identify the immunoreactivity of the recombinant protein, and establish the enzyme linked immunosorbent assay (ELISA) to detect anti-ASGPR antibodies in diagnosis of autoimmune hepatitis. Methods The CRDHI cDNA (435 bp) was subcloned into eukaryotic vector PEGH, and the recombinant protein expression was induced by D (+)-Galactose. The recombinant CRDH1 was purified with Glutathione Sepharose 4B, and its immunoreactivity was identified by SDS-PAGE and western blot as well as MALDI-TOF. ELISA was established to detect the anti-ASGPR antibodies in serum samples of 45 patients with AIH, 30 patients with SLE, 30 patients with RA, 10 patients with SS and 30 normal controls. Results The sequencing of recombinant plasmid showed the CRDH1 gene was successfully inserted to the eukaryotic expression vector with correct sequence and open reading frame. The fusion protein showed a molecular weight of 42 500 Da on SDS-PAGE gel and confirmed to be the human ASGPR by MALDI-MS through peptide mass fingerprint analysis with Mascot in human protein database. It shared 98. 34% homology with ASGPR H1 subunit. Western blot analysis showed that the fusion protein had the same immunoreactivity as human ASGPR. The results of ELISA indicated that the positive rate of anti-ASGPR was 35.6% ( 16/45 ), but the ELISA was negative in other control. There was significant difference of positivity of the autoantibodies between AIH and non-AIH controls (χ2 = 31.85,P < 0. 01 ). Conclusions The human plasmid containing ASGPR is successfully clone into Saccharomyces cerevisiae Y258. The recombinant autoantigen owns good antigenicity and specificity. ELISA established with the purified protein shows good specificity for diagnosis of AIH.

Objective To investigate the association of TGF-β receptor typeⅡ(TβRⅡ)mRNA with lupus nephritis (LN) and disease activity by testing its expression levelin peripheral blood mononuclear cells (PBMCs).Methotis Forty-four patients with LN were included in this study.They were all had active LN.Twepty-eight LN patients were taking glueocorticoids and/or immunosuppressive agents and sixteen had never taken steroids or immunosuppressive agents.The expression levels of T13R H mRNA were semi-quantitativelydetermined by reverse transcription-polymerase chain reaction(RT-PCR).Resuits The expression levels of TβRⅡ mRNA in PBMCs from LN patients(1.7±1.0)were lower than those of non-lupus nephritis(4.0±3.1) and healthy subiects(4.1±2.5),(P<0.01).The difference of the expression levels between patients who took and had never taken glucocorticoids and/or immunosuppressive drugs was significantly statistically(P<0.05).The expression levels of TβRⅡ mRNA in PBMCs of patients with LN were correlated significantly with the systemic lupus erythematosus disease activity index (SLEDAI)scores(r-0.309.P<0.05),titers of anti-dsDNA antibody(r=-0.401,P<0.01)and serum complement C3 level(r=0.621,P<0.01).Conclusion This study suggests that TβRⅡ may be involved in the development of LN,and the TβRⅡ mRNA expression levels in PBMCs from patients with SLE are significantly correlated with LN activity.Glucocortieoids or immunosuppressive drugs can increase the expression levels of TβRⅡ mRNA and ameliorate renal damage.