Objective:To investigate the effect and the mechanism of Apigenin on lipopolysaccharides ( LPS )-induced inflammatory mediators production in murine macrophages. Methods:The murine macrophage cell line RAW 264. 7 cells were cultured in vitro,and were treated with different concentration of Apigenin followed by LPS administration. Expression of heme oxygenase-1 ( HO-1),cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS),phosphorylation of p38 and IκB,nuclear translocation of Nrf2 were detected by Western blot. Production of Nitrite and nitrate ( NOx) was analyzed by colorimetric technique. Secretion of prosta-glandin E2 (PGE2) was detected by ELISA. Activation of NF-κB was measured by luciferase assay. Results: Western blot indicated that apigenin could induce RAW 264. 7 cells expression of HO-1, and pretreatment of SB203580, an inhibitor of p38 significantly inhibited apigenin induced HO-1 expression. In addition,Apigenin could also decrease the content of nuclear transcription factor Nrf2 in cytoplasm and increase its level in the nucleus. Silencing of Nrf2 by specific siRNA could inhibit apigenin-induced HO-1 expression. Furthermore,apigenin administration significantly inhibited LPS-induced NOx production and PGE2 secretion, COX-2 and iNOS expression,IκB phosphorylation and NF-κB activation,and transfection of HO-1 siRNA could reverse these actions. Conclusion:Apigenin inhibits LPS-induced inflammatory response through induction of HO-1 and inhibition of NF-κB in macrophages.

Antimicrobial peptides are a class of small peptides with anti-extrogenous pathogen activities.They are derived from organism and possess antibacterial,antifungus,antiviruses and anticancer cell actions.In recent years,it’s found that some microbial pathogens are able to resist antimicrobial peptides.The constitutive and inducible mechanism of a pathogen resists a given peptide were reviewed in this paper.

Objective To investigate the effect of berberine on insulin resistance induced by free fatty acid in 3T3-L1 adipocytes and the possible molecular mechanism.Methods 3T3-L1 adipocytes were treated with 0.5 mmol/L palmitic acid to induce insulin resistance.Berberine was used for treatment and aspirin for positive control.Glucose oxidase method was employed for measuring the glucose consumption in the medium and 2-deoxy- [~3H]-D-glucose method was used for the determination of glucose uptake.Western blot was used for the determination of IKB kinase(IKK)?SerlS1 phosphorylation,insulin receptor substrance-1(IRS-1)Ser307 phosphorylation,the protein expression of IKK?,IRS-1,phosphatidylinositol 3-kinase(PI-3K)p85 and glucose transporter 4(Glut4).Results After the treatment with 0. 5 mmol/L of palmitic acid for 24 h,glucose consumption by 3T3-L1 adipocytes was decreased by 41%,insulin-stimulated glucose transport was inhibited by 67%,IRS-1 and PI-3K p85 proteins were reduced, and phosphorylations of IKK?Ser181 and IRS-1 Ser307 were induced.The above results were reversed by adding berberine or aspirin.But Glut4 and IKK?protein abundance was not changed during this study.Conclusion Berberine significantly improves insulin resistance induced by free fatty acid in 3T3-L1 adipocytes via inhibiting IKK?serine phosphorylation.

BACKGROUND: This study aimed to investigate the prevalence rate of critical illness-related corticosteroid insufficiency (CIRCI) and the effect of low-dose glucocorticoid on prognosis of CIRCI in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD). METHODS: Since January 2010 to December 2012, 385 patients, who met the criteria of AECOPD, were enrolled in the Intensive Care Unit (ICU) of the First People's Hospital and Municipal Central Hospital of Xiangtan City. The AECOPD patients complicated with CIRCI screened by an adrenalcorticotrophic hormone test within 12 hours after admission to ICU were divided into a treatment group (n=32) and a control group (n=31) for a prospective, randomized and controlled clinical trial. Hydrocortisone (150 mg/d) or normal saline was injected intravenously for 7 days. The patients were followed up for 28 days after injection. The endpoint included 28-day survival time, non-shock time, ICU stay and the period of non-mechanical ventilation. The markers of inflammation C-reactive protein, tumor necrosis factor-α, interleukin 6 and procalcitonin were measured at baseline and 7 days after treatment. The variables were analyzed by Student's t test, the non-parametric statistical test, the Chi-square test or the Kaplan-Meier method with SPSS18.0 statistic software. A P value <0.05 was considered statistically significant. RESULTS: Totally 63 patients were diagnosed with CIRCI by an adrenalcorticotrophic hormone test and the prevalence rate was 16.4％. The shock rate of the AECOPD patients complicated with CIRCI was higher than that of the AECOPD patients without CIRCI (23.8％ vs. 8.7％, P<0.01). Kaplan-Meier analysis revealed that the 28-day survival time of the treatment group was obviously longer than that of the control group (P<0.05). Compared with the control group, shock-free days within 28 days was longer in the treatment group (18.2±9.5 vs. 25.8±4.1, P<0.05). Treatment with low-dose glucocorticoid obviously decreased the markers of infection and inflammation (P<0.01), such as C-reactive protein (13.2±5.5 mg/L vs. 8.3±3.1 mg/L for the control group; 13.5±5.9 mg/L vs. 5.1±2.3 mg/L for the treatment group), tumor necrosis factor-α (26.1±16.2 g/L vs. 17.5±11.7 g/L for the control group; 25.0±14.8 g/L vs. 10.4±7.8 g/L for the treatment group) and procalcitonin (3.88 g/L vs. 2.03 g/L for the control group; 3.77 g/L vs. 1.26 g/L for the treatment group). Furthermore, the markers in the treatment group decreased more obviously than those in the control group (P<0.01). CONCLUSION: The prevalence rate of CIRCI was higher in the patients with AECOPD in the department of critical medicine, and low-dose glucocorticoid treatment for one week reduced the 28-day mortality, shock time and markers of infection and inflammation.

To explore the effect of glycosyl-phosphatidyl inositol-specific phospholipase D (GPI-PLD) on the adhesion function of bone marrow mononuclear cell from patients with myeloid leukemia and analyze its mechanism, the activity of GPI-PLD in bone marrow mononuclear cell from the patients were measured by using GPI-anchored placental alkaline phosphatase (PLAP) as substrate and Triton-X114 partitioning; the adhesion rate and CD24 expression of these cells were measured by MTT and immunohistochemical method respectively, when these cells were or were not treated by 1 mmol/L 1,10-phenanthroline for 5 hours. The results showed that the GPI-PLD activity of bone marrow mononuclear cells from the patients was significantly inhibited after being treated by 1 mmol/L 1, 10-phenanthroline for 5 hours [(42.08 +/- 7.21)% vs (5.4 +/- 2.96)%], while the adhesion rate and the expression of CD24 of these cells were increased [(49.78 +/- 26.73)% vs (61.19 +/- 29.14)%, (16.02 +/- 9.68)% vs (18.5 +/- 11.14)%, respectively)]. It is concluded that depression of GPI-PLD activity can increase the adhesion rate of bone marrow mononuclear cells from the patients while the CD24 expression is enhanced.
Adolescent ; Adult ; Aged ; Bone Marrow Cells ; drug effects ; metabolism ; pathology ; CD24 Antigen ; biosynthesis ; Cell Adhesion ; drug effects ; Cell Survival ; drug effects ; Child ; Female ; Humans ; Immunohistochemistry ; Leukemia, Myeloid ; blood ; Leukocytes, Mononuclear ; drug effects ; metabolism ; pathology ; Male ; Middle Aged ; Phenanthrolines ; pharmacology ; Phospholipase D ; blood ; metabolism

OBJECTIVE: To examine the expression of the inhibitor alpha of nuclear transcription factor kappaB (IkappaBalpha) mRNA expression and its sequence characteristics in human nasopharyngeal carcinoma cell (NPC) lines CNE1, CNE2, HNE1 and HNE2. METHODS: Reverse transcription was performed with the total RNAs isolated from the NPC cell lines CNE1, CNE2, HNE1 and HNE2, as well as the transplanted tumor tissues with HNE1 cells. Then IkappaBalpha cDNA was amplified by PCR, and the products were used to examine IkappaB alpha mRNA expression and DNA sequencing, or the DNA sequencing after the products were cloned into plasmid vector. RESULTS: IkappaB alpha mRNA was expressed in all the 4 nasopharyngeal carcinoma cell lines. DNA sequencing showed that polymorphisms and 5 mutations (A825G, A975G, G576A, A655G and C653A) existed in IkappaB cDNA from the transplanted tumor tissues with HNE1 cells, CNE1 and CNE2 cells. CONCLUSION The expression of IkappaBalpha mRNA not only exists, but DNA polymorphisms and some additional mutations in IkappaBalpha cDNA are also detected in the nasopharyngeal carcinoma cells.
Base Sequence ; Carcinoma ; Cell Line, Tumor ; Humans ; I-kappa B Proteins ; genetics ; NF-KappaB Inhibitor alpha ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; genetics ; Polymorphism, Genetic ; RNA, Messenger ; genetics

Objective To study the neuroprotective effect of erythropoietin(EPO) on neonatal rats model with hypoxia-ischemia encephalopathy(HIE).Methods HIE was induced in rats on 7th day of postnatal age by ligation of right common carotid artery,followed by 2 h of hypoxia(80 mL/L O2).The subjects were divided into sham-operated group,control group and EPO group.EPO 4 000 U/(kg?day) was injected daily from day 2 pre-surgery for 9 to 16 days and PBS was injected in the control group.The neuroprotective effect of EPO on HIE model was detected by brain weight,the difference in weights between the ipsilateral(right) and contralateral(left) brain and the function test.In vitro study,the neural progenitor cell line C17.2 under gone apoptosis following an ischemia-like metabolic inhibition.The effect of EPO on the cell line ischemia modle 17.2 was evaluated by detecting Annexin V with flow cytometry.Results The signi-ficant and sustained brain injury in the hypoxia-ischemia and vehicle-treated group was observed and measured by reduction in relative weights of ipsilateral to contralateral and compromised sensorimotor functions in response to postural reflex test,compared with those of sham-operated animals(Pa

Adult ; Aged ; Coal Mining ; Health Promotion ; methods ; Humans ; Male ; Middle Aged ; Occupational Health ; statistics & numerical data ; Young Adult

OBJECTIVE: To evaluate in vitro microleakage of class V restorations using two self-etching primers with a flowable resin or hybrid resins. METHODS: Forty human molars were divided into 3 groups according to axial surfaces. Each group was randomly assigned to 2 subgroups (n=20) with either butt-joint or the beveled preparations. Class V preparations were cut in cemento-enamel junction. Groups A, B, C were respectively restored with CLB2/Clearfil AP-X, APMB/FHC-Merz or APMB/Liquicoat. Half specimens of each subgroup were thermocycled 2 500 times. After staining, dye penetration was evaluated in the ordinal scale at 40Xmagnifications for occlusal and the gingival margins. The wall adaptation of the randomly selected specimens was analyzed with a SEM on replicas of the sectioned teeth. RESULTS: None of bond systems in this study prevented the microleakge. The restorations with the bur-beveled preparations leaked the same as those with the butt joint preparations (P>0.05). The thermocycled specimens and the non-thermocycled specimens leaked similarly (P>0.05). The flowable resin-liqucoat leaked significantly more than hybrid resin in dentinal margins (P<0.05). CONCLUSION: The self-etching primers can not reduce the microleakge in the dentin-restoration interfaces. The flowable resin leak more than the hybrid resin in dentin-restoration interfaces. The bur- beveled preparations do not significantly reduce the microleakage in class V resin restorations bonded with self-etching primers.