Concanavalin Research Articles

Infection with different Neospora caninum strains causes differences in the glycosylation pattern in the uteri and placentae of Neospora caninum-infected heifers

Neospora caninum is an obligate intracellular parasite that causes abortion in ruminants. Different strains produce differences in the severity of disease outcomes. These differences may cause physiological or pathological changes in cells, modifying the intercellular interactions and intracellular transport pathways that could be evidenced by identifying the terminal sugars. This study aimed to characterize the oligosaccharide pattern in the bovine placenta and uterus after infection with tachyzoites of three different strains of N. caninum (Nc-1, Nc-6 Argentina and Nc Spain-7) during early gestation. Fourteen heifers were inoculated intravenously on day 70 of gestation with 2 × 108 N. caninum tachyzoites and samples of placentae and uteri were analysed by histology and lectin histochemistry. In the infected groups, severe placentitis was associated with changes in lectin binding in the vascular endothelium by Lens culinaris agglutinin (LCA), Pisum sativum agglutinin (PSA) and Ricinus communis I (RCA-I) lectins, in the epithelial cells of the endometrial glands by RCA-I, Dolichos biflorus agglutinin (DBA), succinylated wheat germ agglutinin, peanut agglutinin (PNA), concanavalin-A (CON-A), LCA, PSA and Phaseolus vulgaris erythroagglutinin (PHA-e), and in the trophoblast layer by PNA, CON-A, LCA, PSA, PHA-e, soybean agglutinin, RCA-I, DBA and Bandieraea simplicifolia agglutinin (BSA-I). The results suggest that N. caninum causes changes in the glycosylation pattern in the maternofetal interface tissues and might cause abortions in early gestation due to changes in the cellular structure of the placenta.

Inhibition of RANKL improves the skeletal phenotype of adenine-induced chronic kidney disease in mice.

Skeletal fragility and high fracture rates are common in CKD. A key component of bone loss in CKD with secondary hyperparathyroidism is high bone turnover and cortical bone deterioration through both cortical porosity and cortical thinning. We hypothesized that RANKL drives high bone resorption within cortical bone leading to the development of cortical porosity in CKD (study 1) and that systemic inhibition of RANKL would mitigate the skeletal phenotype of CKD (study 2). In study 1, we assessed the skeletal properties of male and female Dmp1-cre RANKLfl/fl (cKO) and control genotype (Ranklfl/fl; Con) mice after 10wk of adenine-induced CKD (AD; 0.2% dietary adenine). All AD mice regardless of sex or genotype had elevated blood urea nitrogen and high PTH. Con AD mice in both sexes had cortical porosity and lower cortical thickness as well as high osteoclast-covered trabecular surfaces and higher bone formation rate. cKO mice had preserved cortical bone microarchitecture despite high circulating PTH as well as no CKD-induced increases in osteoclasts. In study 2, male mice with established AD CKD were either given a single injection of an anti-RANKL antibody (5mg/kg) 8wk post-induction of CKD or subjected to 3×/wk dosing with risedronate (1.2μg/kg) for 4wk. Anti-RANKL treatment significantly reduced bone formation rate as well as osteoclast surfaces at both trabecular and cortical pore surfaces; risedronate treatment had little effect on these bone parameters. In conclusion, these studies demonstrate that bone-specific RANKL is critical for the development of high bone formation/high osteoclasts and cortical bone loss in CKD with high PTH. Additionally, systemic anti-RANKL ligand therapy in established CKD may help prevent the propagation of cortical bone loss via suppression of bone turnover.

In-situ construction of a NaF-rich cathode–electrolyte interface on Prussian blue toward a 3000-cycle-life sodium-ion battery

As a kind of high-voltage cathode for sodium-ion batteries (SIBs), the instability of a cathode–electrolyte interface (CEI) of Prussian blue (PB) results in the consumption of electrolyte and poor cycling life. Herein, an artificial NaF-rich CEI is in-situ designed on a PB cathode via a chemically presodiated strategy to remarkably prolong the cycling life of SIBs. Based on a spontaneous redox reaction between a chemical presodiation solvent and a polyvinylidene fluoride (PVDF) binder, a homogeneous, thin, and NaF-rich CEI is conformally coated on the surface of PB, forming CEI@PB. The components of CEI within CEI@PB mainly comprise inorganic Na-salts (NaF, Na2CO3, and NaHCO3) and organic Na-based species (R-O-Na, R–CO–Na, and R–O2CO–Na), which can ensure the rapid transport of Na+ ions while effectively preventing CEI@PB against the attack of organic solvent and improving its cycling stability. Besides, density functional theory calculations also reveal that NaF is a good electronic insulator and ionic conductor, which is suitable as a stable CEI. Benefiting from the aforementioned merits, the CEI@PB cathode with a high areal capacity of 0.61 mAh cm−2 delivers an ultra-long cycling life of 3000 cycles. More importantly, the concept of in-situ CEI can be extended to other cathodes beyond Na batteries discussed herein.

Thermal Inactivation of Myrosinase from White Mustard Seeds

Myrosinases (thioglucosidases) catalyze the hydrolysis of a class of compounds called glucosinolates, of which the aglycones show various biological functions. It is often necessary to minimize the loss of myrosinase activity during thermal processing of cruciferous vegetables. Myrosinase was isolated from a popular spice, white mustard (Sinapis alba), and its thermal inactivation kinetics was investigated. The enzyme was extracted from white mustard seeds and purified by a sequential processes of ammonium sulfate fractionation, Concanavalin A-Sepharose column chromatography, and gel permeation chromatography. At least three isozymes were revealed by Concanavalin A-Sepharose column chromatography. The purity of the major myrosinase was examined by native polyacrylamide gel electrophoresis and on-gel activity staining with methyl red. The molecular weight of the major enzyme was estimated to be 171 kDa. When the consecutive step model was used for the thermal inactivation of the major myrosinase, its inactivation energy was 44.388 kJ/mol for the early stage of destruction and 32.019 kJ/mol for the late stage of destruction. When the distinct two enzymes model was used, the inactivation energy was 77.772 kJ/mol for the labile enzyme and 95.145 kJ/mol for the stable enzyme. The thermal inactivation energies lie within energy range causing nutrient destruction on heating.

Concentration-dependent effect of silymarin on concanavalin A-stimulated mouse spleen cells in vitro

AbstractAims: Silymarin (SIL), a mixture of phenolic compounds, has a pleiotropic mode of action on various cell types, including immune cells. In this study, we investigated the concentration-dependent effect of SIL on proliferation of concanavalin A (CoA)-stimulated mouse spleen T lymphocytes, their viability, and secretion of IFN-g and IL-4 cytokinesex vivoin relation to gene expressions of transcription factors nuclear factor kappa B and Foxp3. In addition, metabolic activity of T cells was determined as changes in the mitochondrial membrane potential and apoptosis.Material/Methods: Isolated splenocytes were stimulated with lectin CoA and treated with SIL atthe concentrations of 5, 10, 20, and 40 μg/ml for 70 h and unstimulated cells served as the control. Cultures of splenocytes were evaluated for proliferation index following BrdU incorporation and viability of cells after trypan blue staining. Gene expressions of transcription factors and cytokines were assessed using real-time PCR, whereas ELISA test was applied to measure cytokine secretion. Mitochondrial membrane potential and apoptosis were determined by flow cytometry.Results: We demonstrated that CoA-activated mouse spleen T lymphocytes show different susceptibilities to low (£10 μg/ml) and higher (20 and 40 μg/ml) SIL concentrations. Low concentrations resulted in increased proliferation, cytokine secretion, and mitochondrial membrane potential and reduced transition of cells to apoptosis. High concentration of SIL had the opposite effect without exerting significant cytotoxicity and upregulated genes for cytokines and transcription factors on mRNA level. It is possible that individual subpopulations of T cells induced by CoA were differentially affected by the various SIL concentrations and the dose of 40 μg/ml had the profound suppressive effect. This correlated with the highest expression of Foxp3 factor, indicating that this dose stimulated preferential differentiation to Tregs lymphocytes.Conclusions: Treatment with suitable doses of SIL can provide potential benefits in the modulation of host immune functions in various diseases.

Mechanism of infliximab in preventing autoimmune hepatitis in mice model

Objective To observe the preventive effects of infliximab in autoimmune hepatitis (AIH) and to explore its mechanism. Methods The mice AIH model was established by injecting concanavalin A (Con-A) into the caudal vein. Forty mice were divided into prevention group and control group, with 20 mice in each group. The mice of prevention group were injected intravenously with infliximab (20 mg/kg) one hour before Con-A injection and the mice of control group were administrated with 200 μL phosphate buffered saline (PBS). Serum was collected 3, 8, 12 and 24 h after Con-A/PBS injection. The serum level of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) was detected by colorimetry. The level of cytokine interleukin (IL)-6, IL-1β, interferon gamma (IFN-γ), IL-4, IL-17A, IL-10 and chemokine C-X-C motif ligand 10 (CXCL10) was measured by enzyme-linked immunosorbent assay (ELISA). Liver samples were taken 12 h after Con-A/PBS injection for hematoxylin-eosin staining. Liver infiltrated lymphocytes were assessed by flow cytometry. The expression of T-box transcription factor 21 (TBX21), GATA binding protein 3 (GATA3), RAR related orphan receptor C (RORC) and CXCL10 at mRNA level was evaluated by quantitative real-time polymerase chain reaction (qRT-PCR). The expression of CXCL10 in liver was detected by Western blotting. Paired t test and one-way analysis of variance were used for statistic analysis. Results At 8, 12, and 24 h after Con-A injection, the serum ALT level, AST level, IL-1β and IFN-γ of prevention group were all lower than those of control group ((545.8±190.3) U/L vs. (865.8±237.7) U/L, (947.6±267.9) U/L vs. (1 448.0±403.5) U/L, (508.6±131.1) U/L vs. (976.6±207.6) U/L; (620.7±132.0) U/L vs. (952.9±106.8) U/L, (801.6±212.0) U/L vs. (1 424.8±236.0) U/L, (632.1±117.8) U/L vs. (1 008.3±187.5) U/L; (31.38±10.12) ng/L vs. (48.12±11.53) ng/L, (39.34±11.40) ng/L vs. (60.00±14.17) ng/L, (29.49±8.22) ng/L vs. (46.89±5.50) ng/L; and (432.93±66.82) ng/L vs. (674.66±97.88) ng/L, (655.09±169.17) ng/L vs. (937.90±166.36) ng/L, (263.40±54.97) ng/L vs. (410.74±86.64) ng/L), and the differences were statistically significant (t = 2.350, 2.308, 4.263, 4.374, 4.860, 3.806, 2.440, 2.541, 3.939, 4.560, 2.660 and 3.210; all P 0.05). Conclusions Infliximab has certain preventive effects in mice AIH model, which may be achieved by antagonizing TNF-α and decreasing the expression of CXCL10 in liver, reducing the infiltration of T-helper 1 cells and CD8+ T cells into liver, and by reducing T lymphocyte activation induced by inflammatory cytokines thus alleviating the damage of T lymphocytes to hepatocytes. Key words: Hepatitis, autoimmune; Infliximab; C-X-C motif chemokine ligand 10; Mice

Role of intestinal barrier in the pathogenesis of autoimmune hepatitis

Objective To observe and analyze the role of intestinal barrier in the pathognesis of autoimmune hepatitis (AIH), to explain the pathogenesis of AIH and to explore the intestinal based new treatment strategies. Methods A total of 14 AIH patients from January to December 2017 at Tianjin Medical University General Hospital (six patients without liver cirrhosis, and eight patients with liver cirrhosis) and 10 healthy controls were enrolled. The serum levels of D-lactic acid (D-Lac) and diamine oxidase (DAO) were detected by enzyme-linked immunosorbent assay. Real time fluorescence quantitative polymerase chain reaction was used to detect the relative expression levels of connexin (zonula occluden-1 (ZO-1), occludin), cytokines (interleukin(IL)-2, interferon(IFN)-γ, IL-4, IL-10) and Toll-like receptor 4 (TLR4) in terminal ileal tissues of each group. The relative expression of secretory immunoglobulin A (sIgA) in the terminal ileum was determined by Western blotting. Thirty BALB/c mice were selected and divided into blank control group, dextran sulfate sodium (DSS) group, concanavalin A (ConA) group, DSS+ ConA group, and DSS+ bacterium+ ConA group, with six mice in each group. The relative expression levels of ZO-1, occludin in mouse colonic tissues, serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, and inflammatory activity degree of liver tissues (Knodell score) of each group were measured. T-test and one-way analysis of variance were performed for statistical analysis. Results The serum D-Lac and DAO levels of AIH with liver cirrhosis group and AIH without liver cirrhosis group were both higher than those of healthy control group ((1 768.2±147.1) μg/L, (436.2±197.0) μg/L vs. (100.2±10.9) μg/L, and (11.5±2.5) U/L, (5.4±0.9) U/mL vs. (3.5±0.9) U/mL), and the levels of D-Lac and DAO of AIH with liver cirrhosis group were the highest; and the differences were statistically significant (t=5.512, 36.010, 4.088 and 9.443, F=396.958 and 46.640, all P<0.01). The relative expression levels of ZO-1 and occludin in the terminal ileal mucosa of AIH with liver cirrhosis group were lower than those of healthy control group (0.20±0.14 vs. 1.67±0.51, 0.12±0.09 vs. 0.90±0.21), and the relative expression of ZO-1 in AIH without liver cirrhosis group was lower than that in healthy control group (0.99±0.37 vs. 1.67±0.51); and the differences were statistically significant (t=8.641, 7.407 and 2.295, all P<0.05). The relative expression levels of IL-2 and IFN-γ in terminal ileal tissues of AIH with liver cirrhosis group were higher than those of healthy control group (1.11±0.43 vs. 0.24 ±0.16, and 3.50 ± 1.90 vs. 0.32±0.30), however the relative expression of sIgA in terminal ileal tissues was lower than that of healthy control group (0.506±0.024 vs. 1.081±0.102); and the differences were statistically significant (t=4.679, 3.981 and 5.493, all P<0.05). While the relative expression levels of IL-10 in AIH with liver cirrhosis group and AIH without liver cirrhosis group were lower than that in healthy control group (0.30±0.20, 0.42±0.24 vs. 0.84± 0.23), and the relative expression levels of TLR4 in ileum mucosa of the both groups were higher than that of healthy control group (8.74 ±5.13, 6.74 ±3.65 vs. 0.89 ± 0.70); and the differences were statistically significant (t=3.095, 4.816, 3.856 and 3.685, all P<0.05). The relative expression levels of ZO-1 and occludin of DSS+ ConA group were lower than those of ConA group (0.14±0.08 vs. 0.98±0.13, and 0.09±0.02 vs. 0.98±0.16), however serum ALT, AST levels and the Knodell score were all higher than those of ConA group ((5 496.67±618.83) U/L vs. (3 325.00±1 030.06) U/L, (8 825.00±1 165.35) U/L vs. (5 433.33±1 691.14) U/L, and 18.00±2.00 vs. 9.33±3.01); and the differences were statistically significant (t=13.480, 13.520, 4.227, 4.045 and -2.892, all P<0.05). The relative expression levels of ZO-1 and occludin in DSS+ bacterium+ ConA group were higher than those in DSS+ ConA group (0.46±0.08 vs. 0.14±0.08, and 0.53±0.15 vs. 0.09±0.02), while serum ALT and AST levels were lower than those of DSS+ ConA group ((4 343.33±252.16) U/L vs. (5 496.67±618.83) U/L, and (6 123.33±1 086.60) U/L vs. (8 825.00±1 165.35) U/L); and the differences were statistically significant (t=6.928, 7.122, 4.228 and 4.153, all P<0.01). Conclusions AIH patients have increased intestinal permeability and impaired intestinal barrier which is more serious in patients with liver cirrhosis than in patients without cirrhosis. The intestinal barrier injury can aggravate ConA-induced immune-mediated liver injury. While the protection and repair of intestinal barrier can alleviate immune-mediated liver injury induced by ConA. Key words: Hepatitis, autoimmune; Concanavalin A; Intestinal mucosal barrier; Dextran sulfate sodium; Bifidobacterium

Studies on the immunoregulation of Ginseng radix et rhizoma rubra aqueous extract in mice

Objective To investigate the immune enhancement effect of red ginseng aqueous extract in mice. Methods The aqueous extract of red ginseng was obtained by water extraction and concentration, and the aqueous extract 10 ml was equivalent to raw material 2.86 g. The experiment was divided into the blank control group, low dose group of red ginseng aqueous extract 0.24 g/(kg•d), middle dose group 0.48 g/(kg•d) and high dose group 1.43 g/(kg•d). In order to explore the immunoregulation effect of red ginseng aqueous extract, Organ/body weight ratio measurement, the delayed type hypersensitivity (DTH) reaction, the spleen lymphocyte transformation experiment induced by Con A, the detection of antibody-producing, the determination of serum hemolysin experiments, the clearance rate of carbon particles, the phagocytosis effect of macrophages with chicken red blood cell experiments, and the determination of natural killer cell activity were carried out with the mice. Results Compared to the blank control group, the tow edema degree (0.62 ± 0.14 mm, 0.53 ± 0.12 mm vs. 0.36 ± 0.10 mm) of low dose group and medium dose group significantly increased (P<0.05). The ability of proliferation of T lymphocytes (0.173 ± 0.054, 0.189 ± 0.063 vs. 0.098 ± 0.012) of low dose group and high dose group significantly increased (P<0.05). The number of haemolytic empty spots (137.49×103 ± 24.73×103, 148.43×103 ± 27.53×103vs. 112.96×103 ± 26.28×103) of medium dose group and high dose group significantly increased (P<0.05). The phagocytosis rate (35.67% ± 3.82%, 49.26% ± 6.54%, 57.92% ± 7.36% vs. 24.34% ± 4.22%) and index (0.72 ± 0.23, 0.82 ± 0.15, 0.91 ± 0.26 vs. 0.35 ± 0.11) of low-, medium-, high-dose group significantly increased (P<0.05). However, there was no statistical difference in the organ/body weight ratio of the experimental animals, the determination of serum hemolysin experiments, the clearance rate of carbon particles, and NK cells activity. Conclusion Red ginseng aqueous extract has an exact role in immunity improvement in mice. Key words: Ginseng radix et rhizoma; Ginseng radix et rhizoma rubra; Immunoregulation; Mice

Identification of mesenchymal stem-like cells in cerebral cortex of normal rats

Objective To investigate whether the cells with the characteristic of mesenchymal stem cells (MSCs) could be found in rat cerebral cortex. Methods Rat cerebral cortex samples were digested with typeⅡ collagenase into single cells. Culture medium for MSCs was used for cell culture. The morphology of the cells was observed under a microscope. MSC-specific surface markers were detected by flow cytometry. Differentiation potential favoring adipogenesis or osteogenesis of the subcultured cells was analyzed. Lymphocyte transformation test (LTT) was performed to analyze their immunosuppressive function. Bone marrow MSCs (BM-MSCs) were used as positive control cells in all experiments. Results The fibroblast-like cells isolated from the cerebral cortex of rats were morphologically heomeogeneous after cell culture and grew in a spiral pattern. On the surface of these cells, CD29, CD90 and CD146 were highly expressed at mRNA level, while the expression of CD45 and CD31 at mRNA level were not detected. Moreover, they could be induced to differentiate into adipocytes and osteoblasts. Results of LTT showed that they were functionally equivalent to BM-MSCs in inhibiting the proliferation of spleen lymphocytes induced by concanavalin A. Conclusions This study suggested that the cells with the characteristic of MSCs could be detected in rat cerebral cortex. Key words: Cerebral cortex; Mesenchymal stem cell; Lymphocyte transformation test

Revealing the sodium storage of surface C[dbnd]O structure in high performance Na-ion battery

Although sodium-ion batteries (SIBs) have gotten a growing concern for energy storage system because of the natural sufficient of sodium, their progress is hindered due to destitute insertion/extraction property of Na+ in electrodes. This work reports a clear study of an ultralong cycle stability and high specific capacity in biomass derived carbon (BDC). The Na+ storage mechanism is rely on the reaction between surface CO structure and Na+ (there is no electrode structure change during this reaction). This BDC with more surface CO structure is useful for the Na+ storage, When evaluated as an anode for SIBs, it displays a good rate capacity (514 mA h·g−1 at 0.05 A·g−1, 212 mA h·g−1 at 10 A·g−1) and a high reversible capacity (315 mA h·g−1 at 1 A·g−1 after 900 cycles).

Immunomodulatory effect of schisandrae oil in mouse model of autoimmune hepatitis induced by concanavalin A

Immunomodulatory effect of schisandrae oil in mouse model of autoimmune hepatitis induced by concanavalin A

Effect of TGF-β1 and IL-10 on the Immunoregulatory Function of Extracellular Vesicles Derived from Mesenchymal Stem Cells

To investigate the effect of immune regulatory molecules TGF-β1 and IL-10 on the immunoregulatory activities of extracellular vesicles(EV) secreted from mesenchymal stem cells (MSCs). MSC were isolated from human umbilical cord and expanded, then were treated with TGF-β1 and IL-10 for 72h, and MSC-EVs in supernatants were isolated. The total protein content of all samples was determined by BCA methed. The morphological structure was observed by transmission electron microscopy. The surface markers of MSC-EV were analyzed by flow cytometry. The apoptosis of peripheral blood mononuclear cells(PBMNC) stimulated by ConA and the proportion of CD4+CD25+/CD127- (Treg) cells were detected by flow cytometry after incubation with MSC-EV for 72 h. The CBA and ELISA kit were used to detect the contents of IL2, IL4, IL6, IL10, IFN-γ, TNF-α, Th17A and TGF-β1 in PBMC supernatants and MSC-EV. All the samples showed that the typical cup-shaped membrane-like structure was observed by transmission electron microscopy, and CD9, CD44, CD63 and CD81 expressed. After TGF-β1 treatment, the MSC-EV displayed the strongest ability to promote PBMNC apoptosis（P<0.01）, and in all the samples the proportion of Treg cells increased. MSC-EV could increase the content of IL-10 in the supernatants of PBMNC culture, the content of TGF-β1 in PBMNC supernatants after MSC treatment with TGF-β1 was lower than that in untreated group(P<0.05). The content of IL-6 in MSC-EV increased significantly after treatment with TGF-β1, and the content of TGF-β1 decreased. TGF-β1 alters the immnomodulatory function of MSC-EV and its underlying mechanisms need to be clarified in further investigations.

Disorders of local pulmonary immune response in patients with chronic obstructive pulmonary diseases during exacerbation

Introduction. Morbidity due to reduced local immune reaction in patients affected by chronic obstructive pulmonary disease (COPD) is extremely high and continues to grow. The results of a research on changes in the local immune response in patients with COPD at the exacerbation stage are discussed in this paper. The pathogenesis of COPD during exacerbation is characterized by deterioration of the local innate and acquired immunity. The process is accompanied by pathological changes of the systemic immunity. Various cells of the immune response are involved in the inflammation process, attracting increased amounts of macrophages, neutrophils, T- and B-lymphocytes, and dendritic cells. The number of neutrophils and B-lymphocytes are observed in patients suffering from a severe disorder. Most common symptoms of COPD are irreversible narrowing of the airways, destruction of the parenchyma of the lung accompanied by emphysema, and progression of the obstructive pulmonary disease. The aim of the study – to learn the changes in functionality of bronchoalveolar lavage (BAL) cells and peripheral blood cells (PBC) in COPD patients and provide an explanation for these changes in light of pathogenetic characteristics to predict the course of the disease and justify the necessity of a local immunotherapy in comprehensive treatment of COPD patients. Research Methods. The focus of the research was on BAL cells and PBC of patients with COPD at different stages and on blood serum. CD3, CD4, CD8, CD16 and CD19 of BAL and PBC cells were identified using different specific monoclonal antibodies. Receptor expression to IL-2 (CD25) was examined, adhesive molecules were identified using CD54, proliferation levels were measured using IPO-38, as well as was measured the expression of class II antigens MHC (HLA-DR) in the blast transformation reaction that was induced by PHA and Con A mitogens. Phagocytic activity of phagocytizing neutrophils cells, alveolar macrophages (AM), and phagocytizing PBC cells was evaluated. Levels of free radical processes and antioxidant defence were studied using a chemiluminescence method. Results and Discussion. The amount of AM cells in COPD patients declined significantly, whereas the amount of neutrophils notably increased, as well as concentrations of the myeloperoxidase and the eosinophil cationic protein. Examination of phlegm and BAL cells obtained from COPD patients at the exacerbation stage of the disease indicate that 23 % of samples displayed gram-negative bacterial flora (primary pathogens being Pseudomones aeruginosa, Haemophillus influensae, Enterobacter spp.), 15 % – candidas, 61 % – pathogen type was not identified. Researching local and systemic immune response in COPD patients and functional activity changes of BAL and PBC indicated that patients showed significant changes in their local immune system, which manifested in significant reduction in the amount of neutrophils in the lavage, reduction in CD3, CD19, and increased expression of receptors to HLA-DR antigen expression. Summary. Parallel study of the local and systemic immunological responses contributes to expand our knowledge and understanding of pathogenesis of COPD. It is useful in determining methods to help decrease the number of complications and aid in COPD prophylaxis in persons of working age. It also provides information on which local immunotherapy will prove to be most effective. Exploring the subject further should provide fundamental understanding of similar and different symptoms of various disorders in patients with inflammation processes at the local immune level in different body locations, such as respiratory organs during COPD, in the peritoneal cavity during peritonitis, dermal lesions or any other changes in the local immune system of a different body location.

The hepatoprotective effect of lycopene on Con A-induced liver injury in mice.

Lycopene, the main fat-soluble pigment responsible for the red color of ripe tomatoes, is a symmetrical tetraterpene comprising eight isoprene units. In vitro and in vivo studies have shown that lycopene acts as a potent antioxidant; it is 100 times more effective than vitamin E and 125 times more effective than glutathione as an antioxidant. Here, we divided BALB/c male mice into three equal groups: control, Concanavalin A (Con A), and Con A and lycopene. The control group mice received only vehicle by intraperitoneal injection, the Con A group mice were given Con A, and the Con A and lycopene group mice received Con A and lycopene. The results showed that Con A administration increased histopathological damage, and the levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), interleukin (IL)-6, interferon (IFN)-γ and tumor necrosis factor (TNF)-α were increased in serum samples whereas the levels of these compounds were significantly decreased in the Con A and lycopene group compared to the Con A group. Furthermore, we observed that lycopene led to an increase in cell viability and cell growth. The results of this study revealed that lycopene might be a useful hepatoprotective agent for reducing increased proinflammatory cytokine levels, and for increasing cell viability and cell growth.

La migración en el testimonio indígena de América Latina

The article examines the representation and function of the migration of indigenous people in the Latin American testimonial genre. Specifically, the articles considers the bilingual Mapudungun-Spanish Pascual Cona (1930) and the Quechua-Spanish Gregorio Condori Mamani (1977) of the Mapuche and Quechua cultures respectively. It is argued that the dynamics and consequences of migration to the city are represented in these texts as traumatic experiences that negatively mark the lives of indigenous narrators, since both fail to coexist with the Spanish-speaking hegemonic groups in city contexts.

The role of RAGE in high mobility group box-1 protein1-mediated CD4+ T cells differentiation to Th1/Th2

Objective To explore the role of receptor for advanced glycation end products (RAGE) in HMGB1-mediated CD4+ T cells differentiation to Th1/Th2. Methods CD4+ T lymphocytes isolated from the spleens of male BALB/C mice by magnetic beads were suspended in RPMI-1640 with 10% FCS in 2×106cell/well on 96-well cell culture plates in vitro. The cells were randomly divided 4 groups according to concentration of HMGB1 treatment: control group, 10 ng/mL group, 100 ng/mL group, 1 000 ng/mL group after stimulation with ConA in 3 μg/mL for 12 hours. IL-4 and IFN-γ levels in culture supernatants were quantitated by ELISA kits after HMGB1 stimulation for 12, 24, and 48 h. According to the results, cultured cells were exposed to HMGB1 in 100 ng/mL for 24 h in the following experiments. The cells were randomly divided into 4 groups: control group, A group, B group, C group, and each group were cultured with ConA in 3 μg/mL for 24 h. The cells of control group and other three groups were stimulated with PBS or 100 μg/L HMGB1 for 24 h. The cells of B, C groups were incubated with 1/200 diluted 5 μg/L anti-RAGE Abs (anti-bodies) or PBS for 2 h before HMGB1 stimulation. The cell suspension was obtained to detect the levels of IL-4 and IFN-γ by EILSA and the protein levels and mRNA expressions of RAGE, CATA-3 were detected by western-blot and real-time fluorescent quantitative PCR, respectively. Results Compared with control group, CD4+ T cells incubated with increasing concentrations of HMGB1 (100, 1 000 ng/mL) for 24 h resulted in a decrease in IFN-γ/IL-4 ratio (P<0.05). When CD4+ T cells were exposed to 100 ng/mL HMGB1 for 12 h, IFN-γ/IL-4 ratio was markedly increased. However, CD4+ T cells treated with 100 ng/mL HMGB1 for 24, 48 h, IFN-γ/IL-4 ratio was markedly inhibited (P<0.05). Compared with control group, protein levels and mRNA expressions of RAGE and GATA3 of cells in A group were significantly increased (P<0.05), and IFN-γ/IL-4 ratio of cell suspension in A group and B group was significantly down-regulated (P<0.05). Compared with A group, IFN-γ/IL-4 ratio of cell suspension in C group was increased (P<0.05), and expression of GATA3 mRNA was down-regulated (P<0.05). Compared with A group, protein level of RAGE of cells in C group was significantly down-regulated (P<0.05), but protein level of RAGE of cells in C group was still increased compared with control group (P<0.05). Conclusion Th1/Th2 differentiation induced by HMGB1 on CD4+ T lymphocytes in vitro was at least partly mediated by over-activating RAGE/GATA3 pathway. Key words: High mobility group box-1 protein; Receptor for advanced glycation endproducts; CD4+ T cells; Th1/Th2

The effect of serum amyloid protein A on the hepatitis development induced by concanavalin A

Objective To investigate the effect of serum amyloid A (Saa) on the hepatitis development which induced by concanavalin A (ConA) and the action mechanism. Methods To establish hepatitis mouse model by injecting ConA in Saa over-expression transgenic mice (TG) and wild-type mice (WT), and the expression of monocyte chemotactic protein 1 (MCP1), chemokine 1α (MIP1α), chemokine 1β (MIP1β), chemokine IP10 (IP10) and Eotaxin gene were detected by real-time quantitative polymerase chain reaction (Real-time PCR) in liver tissue. The concentration of alanine transaminase (ALT) and glutamic oxalacetic transaminase (AST) in serum were measured by autoanalyzer. The concentration of tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), interleukin-6 (IL-6) and interleukin-8(IL-8) in serum were measured by enzyme-linked immunosorbent assay (ELISA), and the expression of Saa, p-IκBα(Ser32/36) and IκBα proteins were detected by Western blotting. The isolation and purification of CD4+ T cells in liver and spleen were performed and tested by Flow cytometry. Results The overexpression of MCP1, MIP1α, MIP1β, IP10, Eotaxin gene in TG group (0.61±0.13, 0.89±0.15, 0.76±0.12, 0.37±0.08, 0.98±0.17) were significantly higher than WT group (0.13±0.03, 0.27±0.02, 0.16±0.06, 0.07±0.02, 0.34±0.07), (P=0.001, 0.001, 0.001, 0.021, 0.001, respectively)). At 6 h, the concentration of ALT (16.2±3.1) U/L and AST (18.8±4.2) U/L were significantly higher than WT group (P=0.002, 0.001). The secretion of TNF-α, IFN-γ, IL-6 and IL-8 were higher in serum of TG mice [(813.6±114.2), (64.5±8.9), (1 045.8±138.6), (43.4±5.5) pg/ml] than in WT mice [(253.1±33.5), (10.5±6.7), (374.1±53.7), (24.8±3.4) pg/ml] (P=0.011, 0.006, 0.005, 0.014). Flow cytometry indicated that the rates of CD4+ T cell and F4/80+ CD11b+ macrophage cell in liver of TG group (60.72±3.65)% and (9.86±0.58)% were significantly higher than WT group mice (35.41±2.45)% and (2.73±0.51)%, (P=0.011, 0.014). Western blotting indicated that the levels of p-IκBα (Ser32/36) protein levels significantly increased, while decreased after CU-CPT22 treatment in TG mice liver tissue. Conclusion Saa overexpression promoted hepatitis development by inducing chemokines expression in a Toll like receptors 2 (TLR2) activation-dependent manner, and play a role by increasing the number of CD4+ T cells and macrophages Key words: Saa; Concanavalin A; Chemokine; T cell; Hepatitis

The sympathetic nervous system mechanism of exendin-4 in respect of modulating T cell- mediated immune response to severe burns in mice

Objective To investigate the potential effect of glucagon-like peptide-1 (GLP-1) analogue exendin-4 (Ex-4) on immune function of T lymphocytes via neuroendocrine modulation mechanism in mice following severe burns. Methods Male BALB/C mice were randomly(ramdam number) divided into thermal injury group (n=50) and sham-thermal group (n=30). The thermal injury model was made by exposing the back skin of 15% total body surface area (TBSA) to 95 ℃ water for 7 seconds, while in sham-thermal model the mice were immersed in 37 ℃ water instead. The expression of GLP-1 receptor (GLP-1R) was determined in sorted CD4+ T cells from normal mice by immunofluorescence method. In ex vivo experiment, the mice were sacrificed at 24 h post-burn, then the mononuclear cells (MNC) from spleen were separated from both groups and cultured in RMPI 1640 with 10%FCS (fetal calf serum) in presence of ConA (concanavalin A, 5 μg/mL). Cells were pretreated with catecholamine receptor antagonist propranolol (prop) for 1 hour, followed by consecutive dose of Ex-4 for another 48 h. In in vivo experiment, prop (30 mg/kg) was i. p. injected 30 minutes before thermal injury, then Ex-4 (2.4 nmol/kg) was injected i. p. immediately after scalding. Mice were sacrificed at 6 h and 24 h after thermal injury, then the serum and the spleens were collected. Results GLP-1R was expressed on splenic CD4+ T cells from normal mice. Ex-4 exerted no marked effect on the functions of T cells in terms of proliferation and IL-2 secretion at all doses examined ex vivo, which was not affected by pretreatment with prop. In vivo, T cell functions were suppressed by Ex-4 in thermal mice (P<0.05), but was restored by pretreatment with prop. Regardless of ex vivo or in vivo, Ex-4 could induce T cells switched to Th2 response (P<0.05). Moreover, the Th2 switch by Ex-4 was greatly potentiated by prop intervention in thermal mice in vivo other than ex vivo. Norepinephrine level was increased and epinephrine was decreased by Ex-4 in thermal mice. Both norepinephrine and epinephrine levels were obviously enhanced by pretreatment with prop. Conclusions Ex-4 can inhibit the proliferation and IL-2 secretion of splenic T lymphocytes through the sympathetic nervous system, however, it might induce Th2 switch from Th cells by acting directly on GLP-1R. Key words: Burn injury; Glucagon-like peptide-1; T lymphocyte; Immune response; Sympathetic nervous system

Effects of long noncoding RNA TUG1 on migration and invasion of osteosarcoma cells

Objective To study the effects of TUG1 on migration and invasion of osteosarcoma cells. Methods The osteosarcoma Saos-2 cells were classified into three groups: TUG1 small interfering RNA (siRNA group), blank control group (Con-A), and the negative control-siRNA group (Con-B). The transfection reagent oligofectamine was mixed with TUG1 siRNA and nonsense siRNA, respectively. The TUG1 knocking down effects were examined by real-time polymerase chain reaction (PCR). The migration of osteosarcoma cells was determined by wound healing test, and the invasion ability was studied by Transwell assay. Results As compared with Con-A and Con-B groups, the expression level of TUG1 mRNA was decreased in siRNA group. After osteosarcoma cells were transfected with TUG1 siRNA, the results of wound healing test showed that the space between scratches of Con-A, Con-B and siRNA groups was (326±11) μm, (335±15) μm, and (588±11) μm respectively (F=22.761, P=0.000). The results of the Transwell showed that the membrane cell number in Con-A, Con-B and siRNA groups was (125.0±2.2), (118.0±1.8), and (56.0±1.8) cells respectively (F=173.000, P=0.000). Conclusion TUG1 might be associated with migration and invasion of osteosarcoma cells. Key words: Osteogenic sarcoma; Long noncoding RNA; TUG1; Migration; Invasion

Effects of upregulatioon of Forkhead box protein M1 on migration, invasion and epithelial-mesenchymal transition of human ovarian cancer cell

Objective To explore the effects and mechanism of over-expression of Forkhead box protein M1 (FoxM1) gene on the migration, and invasion and epithelial-mesenchymal transition (EMT) of human ovarian cancer cells. Methods FoxM1 over-expression of the virus vector was constructed. All HO-8910 cells were divided into three groups, the control group (Con-A), Lentiviral vector(LV) control group (Con-B) and over-expression group FoxM1. After three group HO-8910 cells was transfected with phosphate buffer (PBS), Lentiviral vector (LV), and LV-FoxM1, the protein levels of FoxM1 were determined by Western blotting, respectively. The migration and invasion ability was evaluated by wound healing assay and Transwell assay, respectively. The E-Cadherin and N-Cadherin of protein of cancer cells was examined by Western blotting assay. Results The results from Western blotting assay showed that the levels protein of FoxM1 were greatly increased in HO-8910 cancer cells transfected with LV-FoxM1. The results of the wound healing test showed that the migration distance of Con-A, Con-B, and LV-FoxM1 group was(156.80±2.26), (161.60±1.81) and (278.60±2.62) μm, respectively (P<0.01); The results of the Transwell invasion assay showed that the membrane cell number of Con-A, Con-B, and LV-FoxM1 group was (68.56±1.69) and (67.27±1.36) and (126.38±1.56), respectively (P<0.01); Compared with the Con-A, there were E-cadherin protein expression decreased; however, N-cadherin protein increased in HO-8910 cancer cells transfected with LV-FoxM1. Conclusion FoxM1 overexpression can promote human ovarian cancer cell migration and invasion, and its mechanism may be related to the promotion of epithelial-mesenchymal transition. Key words: Ovarian carcinoma; Forkhead box protein M1; Migration; Invasion; Epithelial-mesenchymal transition