AIM: To investigate the effects of lipid-induced insulin resistance on plasma resistin and ghrelin levels in awake rats. METHODS: A hyperinsulinaemic-euglycaemic clamp was established in awake chronically catheterized rats. Two groups of rats were studied either with a 4 h intraarterial infusion of lipid/heparin or saline. Insulin-mediated peripheral and hepatic glucose metabolism was assessed by insulin clamp combined with [~3H]-glucose infusion, and plasma resistin and ghrelin concentrations were examined before and after clamp with ELISA. RESULTS: During clamp, there was a significant increase in plasma free fatty acid (FFA, P

Brain ; blood supply ; Fetus ; blood supply ; Heart Defects, Congenital ; diagnostic imaging ; Humans ; Infant, Newborn ; Middle Cerebral Artery ; diagnostic imaging ; Ultrasonography, Doppler, Color ; Ultrasonography, Prenatal

This study was designed to investigate inhibitory effects and possible mechanisms of snake venom tripeptide (pENW) on platelet adhesion in order to promote the development of a novel anti-platelet therapy. To study the inhibitory effects of pENW on platelet adhesion, washed platelets pre-incubated with pENW (116.5-466.2 μmol x L(-1)) were used to test the ability of platelet adhesion to fibrinogen. Effect of pENW on fibrin clot retraction was also tested. Effect of pENW on platelets viability was tested by MTT assay. Effect of pENW on reactive-oxygen species (ROS) levels of platelet was studied by flow cytometry assay. Calcium mobilization in Fura-2/AM-loaded platelets was monitored with a spectrofluorimeter. Cyclic guanosine monophosphate (cGMP) and cyclic adenosine monophosphate (cAMP), thromboxane A2 (determined as its metabolite thromboxane B2) were measured using enzyme immunoassay kits. Akt, ERK and p38 phosphorylation were tested by Western blot. The results showed that pENW inhibited platelet adhesion and fibrin clot retraction in a concentration-dependent manner without cytotoxicity. Intracellular cGMP and cAMP in both resting and thrombin-activated platelets were increased by pENW. In addition, pENW attenuated intracellular Ca2+ mobilization and TXA2 production in platelets stimulated by thrombin. As shown by Western blot assay, Akt, ERK and p38 phosphorylation in thrombin-induced platelet were attenuated by pENW. However, inhibitory effects of pENW had nothing to do with ROS. Thus, pENW exhibited a significant inhibition on platelet adhesion to fibrinogen, which means pENW could block the first step of thrombosis as while as retard the more stable clot formation. The mechanisms of pENW on inhibition platelet adhesion might be related to instant regulations, such as protein kinases.
Blood Platelets ; drug effects ; Blotting, Western ; Calcium ; metabolism ; Cyclic AMP ; metabolism ; Cyclic GMP ; metabolism ; Flow Cytometry ; Phosphorylation ; Platelet Aggregation ; drug effects ; Reactive Oxygen Species ; metabolism ; Snake Venoms ; chemistry ; Thromboxane A2 ; metabolism ; Thromboxane B2 ; metabolism

Aged ; Esophagus ; Foreign Bodies ; complications ; diagnosis ; Hoarseness ; diagnosis ; etiology ; Humans ; Male

Objective patients with pSS and 30 cases of healthv contrels.The streptavidin immunohistochemical staining was performed to detect the expression and distribution of MMP-9 and CD147 in labial salivary glands.Quantitative analysis was performed by image analysis software-image plus 5.0 at the site of positive expression of MMP-9 and CD147.The correlation between their expression and the infiltrating lymphocyte foci per 4 mm2 of labial gland was analyzed by SPSS software as well as the correlation between the expression of MMP-9 and CD 147 in the salivary glands of patients with pSS.Results MMP-9 was hiKhly expressed in labial salivary glands from 52 patients with pSS and 30 healthy controls,but the expression of MMP-9 in pSS was stronger compared with that of healthy controls(P<0.01).There was a significant positive correlation between lymphocyte foci score and up-regulated expression of MMP-9 in labial salivary glands from 52 patients with pSS(P<0.01).CD147 was highly expressed in labial salivary glands from 52 patients with pSS and 23 healthy controls，but over-expression of CD147 in PSS was more Drominent compared with that of controls(P<0.01).There was a significant positive correlation between lymphocyte foci score and up-regulated expression of CD147 in labial salivary glands from 52 patients with pSS(P<0.01).The expression of MMP-9 and CDl47 was detected in ductal and acinar epithelial cells,lymphocyte foci in pSS.There was linear correlation between the expression of MMP-9 and CDl47 in the salivary glands of patients with pSS(P<0.01).Conclusion The results of this study suggest that the abnormal expression of MMP-9 and CD 147 is involved in the pathogenesis of pSS and play a crucial role.The interaction of MMP-9 and CD147 may be one of theimportant mechanisms leading to labial salivary glands destruction found in pSS.

The five flavours of traditional Chinese medicine(TCM)stand for five tastes,such as pungent,sweet,sour,bitter and salt.The five flavours not only stand for the actual tastiness of herbs,but also the effect characteristic of TCM according to traditional medical theory.So this kind of situation led to a disorder of the marking standardization of five flavours.So the authors think that we should mark herbs(or TCM)based on their true tastiness,and this way maybe help us to solve the problem and to research the relationship between the flavours and the effect of the herbs(or TCM).

Endothelial dysfunction is implicated in a variety of cardiovascular diseases although the detailed mechanisms are not yet completely understood. A relationship has been suggested to exist between inflammation and endothelial dysfunction. TNF-α serves as one of the most important pro-inflammatory cytokines. The main objectives of the present study were to explore the effect of PKC-ζ on TNF-α-impaired endothelial function as well as the underlying mechanisms. Acetylcholine-induced endothelium-dependent vasodilation of mouse thoracic aorta stimulated by TNF-α was initially determined. PKC-ζ deficient mice and the specific inhibitor of NADPH oxidase were respectively applied to elucidate their roles in TNF-α-induced endothelial dysfunction. In vitro superoxide generation in HAECs was detected by DHE staining after administration of TNF-α. Meanwhile, the regulatory p47(phox) subunit of NADPH oxidase was evaluated by Western blotting and RT-PCR. The results showed that TNF-α conspicuously impaired endothelium-dependent vasodilation and the impairment was attenuated by either depleting PKC-ζ or inhibiting NADPH oxidase. In vitro TNF-α increased superoxide production and p47(phox) expression in HAECs, and such increases could be ameliorated by the specific PKC-ζ inhibitor. Our findings suggest that superoxide over-production triggered by PKC-ζ-dependent NADPH oxidase activation contributes to TNF-α-induced endothelial dysfunction.

Objective To investigate the expression and role of costimulatory molecule 4-1BB on T cells of patients with rheumatoid arthritis(RA).Metheds The expression of 4-1BB on T lymphocytes from 30 RA patients and 20 healthy controls were detected by flow cytometry.Results The expression of 4-1BB on CD4~+T and CD8~+T lymphocytes from RA patients was significantly higher than that of normal control(18.56?4.08,10.33?2.13 vs 1.24?0.12,0.87?0.09,P＜0.01).There was more expression of 4-1BB on CD4~+T and CD8~+ T lymphocytes stimulated by anti-CD3 antibody from RA patients(33?4 vs 21?8,P＜0.01).In addition,the ra- tio of CD4~+T/CD8+T in RA patients was higher than that of normal controls and was positively correlated with 4-1BB~+CD4~+T cell.The expression of 4-1BB on CD4~+T in RA patients was positively correlated with the level of ESR and IgA(r=0.476,P＜0.05;r=0.659,P＜0.05).Conclusion The costimulatory molecule 4-1BB is ab- normally expressed in T lymphocytes from patients with rheumatoid arthritis.The abnormal expression of 4-1BB on T lymphocytes may play an important role in the development of RA.The expression of 4-1BB on CD4~+T cell may take part in the inflammation of RA.

Objective:To optimize and establish the experimental methods for the determination of 2,4-dihydroxybenzophenone (BP-1) in mouse brain. Methods:BP-1 was determined by high performance liquid chromatography (HPLC) and separated by Waters Symmetry? C18 (4. 6 mm × 250 mm, 5 μm) using isocratic elution, and the sample preparation conditions were optimized by orthogonal experiment design. The mobile phase was methanol-water (volume ratio 3∶1) containing 3% (volume fraction) ace-tic acid (pH 3. 40) at a flow rate of 1. 0 mL/min, and ultraviolet (UV) detection wavelength was set at 290 nm. Retention time was used for qualitative analysis and internal standard method for quantitative analysis. Results: Under the optimized experimental conditions, the calibration curve was linear with a correlation coefficient of 0. 999 8 over the concentration range of 0. 2-10. 0 mg/L. The recoveries of BP-1 were between 96. 8% and 104. 5%. The intra-day and inter-day precision of BP-1 were 3. 5% -5. 7% and 4. 5% -6. 4%, respectively. The extraction recoveries of BP-1 at three concentrations (0. 5, 2. 0, 8. 0 mg/L) in the mouse brain were 90. 5%, 89. 5%, and 97. 7%, and the matrix effect of BP-1 at these three concentrations were 102. 9%, 102. 7%, and 90. 9%, respectively. Conclusion:The method is simple, ac-curate, and suitable for determination of the contents of BP-1 in mouse brain.

Objective To establish a malignant transformed human fetal osteoblastic human cells from the immortalized cell line hFOB1.19 in order to explore the molecular mechanism in tumorigenesis of osteosarcoma. Methods hFOB1.19 cells were treated sequentially by an initiated factor, N-methyl-N’-nitro-N-nitrosoguanidine (MNNG) and a promoter, 12-O-tetradecanoyl phorbol-13-acetate (TPA). The features of malignancy of transformed cells were identified by cell morphology, DNA content analysis, colony forming frequency on soft agar and tumorigenetic test in nude mice. Results Continuous passaging after the treatments resulted in the formation of a few paramorph foci and exhibited in an extensively random orientation. The poly-ploid of DNA was 81.08% in experiment group, much higher than that in control group (55.03%). Compared with that of negative control cell, colony formation efficiency of transformed cells in semisolid agar showed a a significant increase. The transformed cells formed tumors subcutaneously in the nude mice, which were verified to be poorly differentiated osteosarcoma histopathological examination. Conclusion Mocking the process of malignant transformation of human cells, we establish malignant transformed immortalized human fetal osteoblastic human cells (hFOB1.19) model.