Objective To investigate the effect of ajoene on thrombosis and explore the possible mechanism.Method SD rats were randomly divided into five different groups and administered ajoene in 7d.The effect of ajoene on thrombosis in rats was observed with the thrombogenesis model of artery-vein bypass.The serum level of vascular cell adhesion molecule-1(VCAM-1) and P-selectin(Ps)were measured.To observe the effect of ajoene on platelet aggregation,arachidonic acid was used to induce platelet aggregation and platelet function was evaluated by turbidimetry in rabbit.Results Ajoene significantly inhibited the the thrombosis of artery-vein bypass in rats and reduced rate of platelet maximal aggregation and decreased the P-selectin concentration in serum at 25,50,75mg/kg (P

AIM: To investigate the inhibitory effect s of a synthetic CRE-transcription factor decoy oligodeoxynucleotide (CRE-decoy ODN) on the upregulation of the expression of cholecystokinin (CCK) and fosB mRN A induced by chronic morphine administration in SK-N-SH cells. METHODS: The CRE cis-element, TGACGTCA, was palindromic, a sy nthetic single-stranded phosphorothioate oligodeoxynucleotide composed of the CR E sequence self-hybridizes to form a duplex/hairpin. The CRE-palindromic decoy a nd control oligodeoxynucleotides were added to the medium (1 h before exposure t o morphine) at 150 nmol/L in the presence of cationic lipid DOTAP. After the cel ls were treated with 100 ?mol/L morphine for 48 h, 10 ?mol/L naloxone was use d for 15 min. The effects of CRE-decoy ODN on the DNA-binding activity of CREB, the expression of CCK and fosB mRNA were detected by electrophoresis mobi lity shift assay (EMSA) and RT-PCR, respectively. The stability of cell-incorpo rated [ 32P]-labeled CRE-decoy ODN was extracted with phenol:chloroform a nd then subjected to 20% nondenaturing polyacrylamide gel electrophoresis and au toradiography. RESULTS: Chronic morphine administration and acute naloxone-prec ipitated withdrawal significantly activated the DNA-binding activity of CREB and the expression of CCK and fosB mRNA in SK-N-SH cells. The CRE-decoy ODN pen etrated into the cells, specifically downregulated these indexes. CONCLUSIONS: CRE-decoy ODN can significantly downregulates the e xpre ssion of CCK and fosB mRNA through specifically suppressing the DNA-binding activity of CREB activated by chronic morphine administration in SK-N-SH cells.

Objective To investigate the role of nuclear factor-kappa B(NF-?B)in the modulation of diallyl trisulfide(DATS)on interleukin-1?(IL-1?)expression induced by lipopolysaccharide(LPS)in mice with acute lung injury(ALI).Methods Mice were randomly divided into Control group,ALI group,DATS group,DATS prevention group and DATS treatment group.The expression of IL-1? mRNA in the lung tissue was detected by reverse transcription PCR(RT-PCR).NF-?B activity in the lung tissue was detected by electrophoresis mobility shift assay(EMSA).The expression of phospho-I?B and I?B were assayed by Western blot.Results The expression of IL-1? mRNA,NF-?B activity and the phospho-I?B expression in lung tissues increased significantly at ALI group(P

Aim To investigate the effects of cholecystokinin-octopeptide on IL-12 secreted in murine bone marrow-derived dendritic cells induced by lipopolysaccharide.Methods The CCK receptor subtypes were investigated by immunofluorescence in murine bone marrow-derived dendritic cells.Enzyme linked immunosorbent assay and Western blot were used to estimate the contents of IL-12 and p38MAPK activity.Results CCK-1R and CCK-2R were detected in BM-DC;CCK-8(at concentrations of 10-10,10-8,10-6 mol?L-1)could significantly increase the secretion of IL-12 in the LPS-induced DC, and LPS-activated p38MAPK activity in a dose-dependent manner.The effect of CCK-8 was reduced partially by CR1409(a CCK-1R antagonist) and CR2945(a CCK-2R antagonist).Conclusion CCK-8 could dose-dependently increase the expressions of IL-12 in LPS-induced DC,probably by promoting p38MAPK activity and the effect was mediated by CCK1 and CCK2 receptors.

AIM:To observe the effects of cholecystokinin octapeptide(CCK-8) on the expression of proinflammatory cytokines IL-1?,IL-6 and anti-inflammatory cytokines IL-10,IL-4 in LPS-attacked mice.METHODS: Kunming mice were randomly assigned and injected intraperitoneally with LPS alone or/and CCK-8 at different time points.The expression of IL-1?,IL-6,IL-10 and IL-4 in the serum and lung tissues were assayed by ELISA and RT-PCR.RESULTS: The expression of IL-1?,IL-6,IL-10 and IL-4 were upregulated in LPS-attacked mice.Pre-treatment of CCK-8 decreased both IL-1? and IL-6 expression and augmented IL-10 and IL-4 expression in LPS-attacked mice.CONCLUSIONS: CCK-8 exerts an anti-inflammatory effect by inhibiting the expression of IL-1?,IL-6 and increasing the expression of IL-10,IL-4 in LPS-attacked mice,which could alleviate the inflammatory response in lung tissue.

Objective:To construct a human Fab phage antibody library and to obtain some recombinant clones which can express Fab fragment antibody against ? 1-adrenergic receptor.Methods:Fd heavy chain gene and ??? light chains gene of IgG obtained by RT-PCR from peripheral lymphocytes of DCM patients whose anti ? 1-AR antibodies are present were cloned into pComb3 vector and the human Fab phage antibody library was constructed.The library was panned by phage display technology with ? 1-ARECⅡ as antigen.Results:A human Fab phage antibody library with 1.4?10 6 capability was constructed successfully,a positive clone against human ? 1-AR was screened from the phage antibody library.Conclusion:The recombinant clones which express Fab antibody against ? 1-AR can be obtained by phage display technology.

AIM: To investigate in vitro effects of cholecystokinin octapeptide(CCK-8) on the expressions of B7.1 and B7.2 and the costimulatory activity of T lymphocytes in unstimulated macrophages.METHODS: Mouse peritoneal macrophages were isolated and incubated with CCK-8(10-12-10-6 mol/L) for indicated time.The B7.1 and B7.2 expressions of murine peritoneal macrophages were analyzed by flow cytometry.CD4+T cells were isolated from mouse spleen using immunomagnetic beads,and cultured with 1/4 numbers of macrophages which were pretreated with CCK-8 and/or anti-B7.1 antibody,anti-B7.2 antibody,CCK1R antagonist CR1409,CCK2R antagonist CR2945 for 24 h.ConA was added into the culture medium to stimulate CD4+T cell proliferation.The proliferation was determined by measuring -TdR incorporation in a ?-scintillation counter.RESULTS: B7.1 and B7.2 expressions and costimulatory activity of peritoneal macrophages were enhanced by CCK-8 in a dose-dependent manner,and the maximal effects occurred at the concentrations of 10-9 mol/L to 10-7 mol/L.Anti-B7.2 antibody,but not anti-B7.1 antibody,reduced the modulatory role of CCK-8 on costimulatory activity.Both CR1409 and CR2945 reversed the effect of CCK-8 on costimulation,and the role of CR1409 was more significant.CONCLUSION: CCK-8 enhances macrophage costimulatory activity by upregulating B7.2 expression,which is mediated by CCK1R and CCK2R.CCK1R might be the major receptor responsible for the modulation of CCK-8 on costimulation.

Objective To investigate the impact of blood glucose level on the prognosis of breast cancer patients with type 2 diabetes mellitus. Methods Clinical data of 416 breast cancer patients with type 2 diabetes mellitus in Tianjin Medical University Cancer Institute and Hospital from January 2005 to December 2009 were analyzed retrospectively. Patients were divided into 2 groups according to the postprandial 2 hours of blood glucose level (＜11.1 mmol/L and ≥11.1 mmol/L). Clinicopathological characteristics and survival of the two groups were compared. Results There were 310 cases of postprandial 2 hours of blood glucose level (＜11.1 mmol/L), and 106 cases of postprandial 2 hours of blood glucose level (≥11.1 mmol/L) in 416 patients. There was no statistical difference in the clinicopathological features between the two groups (both P＞ 0.05). Univariate analysis showed that the 5-year disease-free survival rate was 81.6 % (253/310) and 67.0 % (71/106) for patients with postprandial 2 hours of blood glucose level＜11.1 mmol/L and postprandial 2 hours of blood glucose level ≥11.1 mmol/L, respectively (χ2= 8.575, P=0.003). The 5-year overall survival rate was 87.7 % (272/310) and 75.5 % (80/106) for patients with postprandial 2 hours of blood glucose level ＜11.1 mmol/L and postprandial 2 hours of blood glucose level≥11.1 mmol/L, respectively (χ2=8.722, P =0.003). Multivariate analysis showed that 2 hours of blood glucose level≥11.1 mmol/L was an independent risk factor of 5-year disease-free survival rate ( HR=1.701, 95 % CI 1.110-2.604, P= 0.015) and 5-year overall survival rate ( HR= 1.972, 95 % CI 1.186-3.279, P= 0.009) for diabetic breast cancer patients. Conclusion The poor control of blood glucose level for the breast cancer patients complicated with type 2 diabetes is related with the increased risk of 5-year recurrence and death, which requires to strictly control the blood glucose level for improving the prognosis of type 2 diabetic breast cancer patients.

Objective To survey the resistance profile of clinical isolates to antibiotics across the hospitals in Dongguan,Guangdong Province during 2015.Methods Kirby-Bauer method or automated system was used to test the susceptibility of clinical isolates to selected antimicrobial agents.Results were analyzed according to CLSI 2015 breakpoints.The susceptibility data were analyzed using WHONET 5.6 software.Results A total of 29 665 strains of microorganisms were isolated,of which gram positive cocci accounted for 32.1％ (9 509/29 665) and gram negative bacilli accounted for 67.9％ (20 156/29 665),respectively.The prevalence of methicillinresistant Staphylococcus was 23.3％ (705/3 024) in S.aureus and 43.6％ (1 054/2 419) in coagulase-negative Staphylococcus.No vancomycin-resistant staphylococcal strain was found.ESBLs-producing strains accounted for 36.4％ (2 554/7 020) in E.coli and 24.5％(792/3 227) in Klebsiella isolates.The prevalence of carbapenem-resistant Enterobacteriaceae was 0.2％ (30/13 077).The prevalence of carbapenem-resistant Pseudomonas aeruginosa (CRPA) and carbapenem-resistant Acinetobacter baumannii (CRAB) was 16.0％ (500/3 116) and 53.9％ (827/1 533),respectively.The prevalence of penicillin-resistant S.pneumoniae (PRSP) strains was 10.1％ (142/1 404).Beta-lactamase was produced in 30.6％ (276/902) of the H.influenzae strains.The prevalence of vancomycin-resistant Enterococcus (VRE) strains was 0.7％ (10/1 441).Conclusions Periodic surveillance of antimicrobial resistance is valuable for rational antimicrobial therapy,formulation of treatment guidelines and infection control and prevention measures,as well as preventing the spread of drug-resistant strains.

AIM: To study the effects of cholecystokinin octapeptide (CCK-8) on T-lymphocyte subsets in keyhole limpet haemocyanin (KLH)-immunized mice.METHODS: Female BALB/c mice were immunized with KLH and injected with different dosages of CCK-8 or saline simultaneously. Positive CD4+and CD8+ T cells in peripheral blood or splenocytes were measured by flow cytometry. The production and mRNA level of Th1 cytokine, IFN-? and Th2 cytokine, IL-4 in the splenocytes were evaluated by ELISA and RT-PCR, respectively. In addition, lung tissue sections were made with HE staining.RESULTS: CCK-8 downregulated the positive CD4+and CD8+ T lymphocytes both in peripheral blood and in splenocytes in KLH-immunized mice, resulting in the reduction of CD4+/CD8+ ratio.CCK-8 improved the production of IFN-?, elevated IFN-? gene transcription, whereas cut down the production of IL-4, decreased IL-4 gene transcription. CCK-8 lightened the pulmonary inflammation induced by KLH.CONCLUSION: CCK-8 inhibits CD4+T lymphocytes activation, Th2 cytokine mRNA expression and protein production in KLH-immunized mice, indicating that CCK-8 modulates adaptive immune response.