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

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

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

ObjectiveTo analyze the causes of discomfort occurring in the interventional treatment of diabetic foot, to discuss the individualized nursing measures for improving the patient's comfort and cooperation,and for increasing the successful rate of the interventional procedure.MethodsThe control group included 9 patients who received conventional nursing care.The study group had 13 patients who accepted individualized nursing care and nursing intervention, which was designed according to every patient's individual conditions. ResultsIn the control group, one patient could not endure the surgery to the end because of the long operating time. Another two patients had to take examinations repeatedly because the imaging quality was very poor,which was caused by the body movement due to uncomfortable mechanical stimulation.Urinary retention occurred in one patient.The mean operative time of the control group was 2.8 hour per surgery.The average dosage of contrast medium used was 150 -300 ml per procedure.All the subjects in the study group completed the surgery successfully,the mean operative time was 2.2 hour per surgery and the average dosage of contrast medium used was 100 -200 ml per procedure.Conclusion During the perioperative period of interventional treatment for diabetic foot, the effective individualized nursing care and nursing intervention,the measures to improve patient's comfort and the cooperation,etc. can certainly increase the successful rate,saving the operating time and reducing the contrast dosage.

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.

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.

Colon ; pathology ; Colonic Polyps ; pathology ; Diagnosis, Differential ; Female ; Humans ; Intestinal Mucosa ; pathology ; Male ; Middle Aged

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.

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.