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Background & objectives: To examine ?-D-mannuronic acid (M2000) effects on L-selectin shedding and leucocyte function-associated antigen-1 (LFA-1) expression as mechanisms of action of this drug in patients with ankylosing spondylitis (AS). Methods: To investigate the molecular consequences of ?-D-mannuronic acid on L-selectin shedding, flow cytometry method was used. Furthermore, the effect of it on LFA-1 gene expression was analyzed by using quantitative real time (qRT)-PCR technique. Results: The LFA-1 expression in patients with AS was higher than controls (P=0.046). The LFA-1 expression after 12 wk therapy with ?-D-mannuronic acid was meaningfully decreased (P=0.01). After 12 wk treatment with ?-D-mannuronic acid, the frequency of CD62L-expressing CD4+ T cells in patients with AS, was not considerably altered, compared to the patients before therapy (P=0.5). Furthermore, after 12 wk therapy with ?-D-mannuronic acid, L-selectin expression levels on CD4+ T-cells in patients with AS, were not remarkably changed, compared to the expression levels of these in patients before treatment (P=0.2). Interpretation & conclusions: The results of this study for the first time showed that ?-D-mannuronic acid can affect events of adhesion cascade in patients with AS. Moreover, ?-D-mannuronic acid presented as an acceptable benefit to AS patients and could aid in the process of disease management
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L-Selectin (SELL) linked with innate immune mechanism involved in neutrophil migration through endothelium to the site of inflammation. Early recruitment of neutrophils at the site of infection is essential to counteract infection at the earliest in conditions like sub-clinical mastitis (SCM). The present study was framed to evaluate the expression pattern of SELL in naturally occurring SCM in crossbred animals using real time PCR technique. Analysis of data on total leukocyte count indicated leucocytosis condition in SCM affected crossbred cows. Relative expression of SELL on peripheral blood leukocytes revealed a significant 3.16 folds down regulation in SCM affected cows when compared with healthy crossbred cows (P<0.05). The possible reason for leukocytosis in SCM animals might be due to down regulation of SELL on leukocytes reducing their ability to transmigrate through blood vessel to infection site. The result of present study revealed a definite role of SELL in SCM which could be explored for therapeutic aspects in near future
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Objective To investigate the effects of recombinant human growth hormone (rhGH) on the expression of CD47, L-selective action hormone (L-selectin), and advanced oxidation protein products (AOPPs) in elderly patients with multiple organ dysfunction syndrome (MODSE). Methods Sixty-six MODSE patients were randomly and equally divided into two groups (33 each): control group (received conventional treatment only) and rhGH treatment group (received conventional and rhGH treatment). Venous blood was taken from all the subjects before treatment and 3rd, 14th and 28th day after treatment. Serum CD47 was detected by flow cytometry, L-selectin and AOPPs concentrations were determined with ELISA and spectrophotometry. The changes in APACHE HI score in the two groups were also observed. Results Compared with that before treatment, the positive rate of CD47 was increased, meanwhile the AOPPs concentrations, L-selectin levels and APACHE HI scores declined markedly in both groups after treatment (P<0.05). Compared with that in control group, the positive rate of CD47 increased more significantly, while the AOPPs concentrations, L-selectin levels, APACHE HI scores and mortalities declined more significantly in rhGH treatment group (P<0.05). In the rhGH treatment group, the positive rate of CD47 increased, the AOPPs concentrations, while L-selectin levels and APACHE III scores declined markedly on the 14th day compared with those on the 3rd day, and on the 28th day as compared with that on the 14th day after treatment (P<0.05). ConclusionrhGH may enhance the immune function and reduce oxidative stress, inflammatory reaction, and mortality in elderly patients with MODSE.
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Background and purpose:Colorectal cancer is a kind of common digestive malignancies, which seriously threaten the human health. Although modern diagnostic and treatment technology has developed rapidly, the incidence and mortality of colorectal cancer continue to show an increasing tendency in recent years, and early diagnosis and prevention of colorectal cancer liver metastases are important to increase the survival rate of patients and to improve the prognosis. P-selectin and L-selectin is attracting much attention in cancer research ifeld recently, and the change of their expression and mediated adhesion play an important role in tumor cell metastasis, but their relationship between the occurrence and clinical stage of colorectal cancer rarely reported. This study aimed to explore whether the serum P-selectin and L-selectin levels of colorectal cancer patients were correlated with clinical and pathological features and the situation before and after surgery.Methods:A total number of 132 cases of colorectal cancer patients and 100 healthy subjects with gender and age-matched were enrolled. Enzyme-linked immunosorbent assay (ELISA) was used to detect serum P-selectin and L-selectin concentrations in 132 cases before and after surgery, 100 healthy subjects were enrolled as the control group, the P-selectin and L-selectin levels were analyzed combined with theclinical and pathological features.Results:Serum P-selectin and L-selectin levels in patients with colorectal cancer before surgery were signiifcantly higher than those in the healthy control group (75.2±13.3vs 26.6±9.6, 89.2±12.7vs 33.9±8.3,P0.05); however, significantly decreased after 3 months (34.8±10.4vs 75.2±13.3, 40.1±9.5vs 89.2±12.7,P0.05).Conclusion:Serum P-selectin and L-selectin levels to some extent relfect the invasion of colorectal cancer, the degree of inifltration and lymph node metastasis, and which can be an important indicator in the development and prognosis of colorectal cancer.
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Reactive oxygen species (ROS) production is one of the main mechanisms used to kill microbes during innate immune response. D-lactic acid, which is augmented during acute ruminal acidosis, reduces platelet activating factor (PAF)-induced ROS production and L-selectin shedding in bovine neutrophils in vitro. This study was conducted to investigate whether acute ruminal acidosis induced by acute oligofructose overload in heifers interferes with ROS production and L-selectin shedding in blood neutrophils. Blood neutrophils and plasma were obtained by jugular venipuncture, while ruminal samples were collected using rumenocentesis. Lactic acid from plasma and ruminal samples was measured by HPLC. PAF-induced ROS production and L-selectin shedding were measured in vitro in bovine neutrophils by a luminol chemiluminescence assay and flow cytometry, respectively. A significant increase in ruminal and plasma lactic acid was recorded in these animals. Specifically, a decrease in PAF-induced ROS production was observed 8 h after oligofructose overload, and this was sustained until 48 h post oligofructose overload. A reduction in PAF-induced L-selectin shedding was observed at 16 h and 32 h post oligofructose overload. Overall, the results indicated that neutrophil PAF responses were altered in heifers with ruminal acidosis, suggesting a potential dysfunction of the innate immune response.
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Animals , Cattle , Female , Acidosis/chemically induced , Blood , Cattle Diseases/chemically induced , Flow Cytometry/veterinary , Immunity, Innate , L-Selectin/metabolism , Neutrophils/drug effects , Oligosaccharides/pharmacology , Platelet Activating Factor/pharmacology , Reactive Oxygen Species/metabolism , RumenABSTRACT
Objective To explore the changes ofintegrin α4β7 and L-selectin levels and the possible relationship between the changes and intestinal mucosal immune function in critical children with gastrointestinal dysfunction.Methods Forty-eight admitted critical children were divided into non-gastrointestinal dysfunction group (29 cases) and gastrointestinal dysfunction group (19 cases),and also compared with 25 healthy people (control group).The expression of integrin α 4β 7 and L-selectin was measured by flow cytometry.Results The expression of integrin α 4β 7 in gastrointestinal dysfunction group [(11.11 ± 1.15)％] was lower than that in non-gastrointestinal dysfunction group [(15.36 ± 1.26)％]and control group [(19.62 ±0.89)％] (P ＜0.05),but there was no significant difference between nongastrointestinal dysfunction group and control group(P ＞ 0.05).The expression of L-selectin in gastrointestinal dysfunction group [(60.68 ±3.72)％] was lower than that in non-gastrointestinal dysfunction group [(69.43 ± 1.52)％] and control group [(88.65 ± 2.41)％] (P ＜ 0.05),and there was significant difference between non-gastrointestinal dysfunction group and control group (P ＜ 0.05).Conclusion The expression of integrin α 4β 7 and L-selectin is decreased at the early stage of gastrointestinal dysfunction in critical childern,and integrin α 4 β 7 is decreased sharply.
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Objective To investigate the effect of L-selectin in experimental allergic encephalomyelitis (EAE) of Wistar rats.Methods The rats were randomly divided into four groups;the normal group,the CFA group, the LMS group and the model group;The animal models were established in rats by immunization with myelin basic protein of spinal cord of guinea pig and complete Freund's adjuvant(CFA).The symptom of EAE was observed; pathological feature and myelin of brain and spinal were detected with HE stain and Loyez's stain respectively.The number of positive vessels of L-selectin expression was detected by immunohistochemistry.Results 100% experimental Wistar rats treated with MBP and levamisole developed EAE,but none of the other groups.The number of positive vessels of L-selectin expression was (31.86 ± 1.39) in model group, obviously higher than that of in the normal group (1.38 ±0.18) ,the CFA group( 1.50 ±0.27) and the LMS group(7.25 ±0.59) (all P <0.05) ;The inflammation cells were found around vessels and demyelination were seen in white matter of brain and spinal cords.Conclusion The expression of L-selectin should play an important role in EAE.
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Objective To investigate the role of PKCθ signal pathway on regulation of L-selectin (CD62L) expression in human activated γδT cells. Methods Human peripheral blood mononuclear cells (PBMC) were stimulated with Mycobacterium tuberculosis antigen (Mtb-Ag) and cultured for 6-8 d to generate Mtb-Ag activated T cells(MtbAT) as γδT cells enrichment T cell line. The MtbAT were stimulated with PMA or PMA + IMN for 3, 6, 12 and 24 h, respectively, or MtbAT cultured for 8 d were stimulated with Mtb-Ag, or PMA, with or without PKC0 inhibitor Rottlerin for 4 h. After the treated cells stained with fluo-rescent labeled monoclonal antibodies, the expression of CD62L on γδT cells were measured by flow cytome-try (FCM ). Results The expression of CD62L on γδT cells cultured for 6-8 d were 75.0%-87.0%. Decrease of CD62L from the surface of γδT cells by 3 h to 12 h after exposure to PMA (42.3% to 23.5% ), but CD62L expression increased to 53.2% when γδT cells were exposed to PMA for 24 h. The expression of CD62L of γδT cells decreased to 52.1% and 39.3% respectively when γδT cells were exposed to PMA + IMN for 3 h and 6 h. After treated with PMA + IMN for 12 h and 24 h, the expression of CD62L were 52.9% and 35. 3% respectively. The CD62L expression of γδT cells treated with PMA and Rottlerin (47.9%) were higher than that treated with PMA alone (31.8%). After Mtb-Ag restimulated MtbAT for 4 h, the CD62L level of γδT cells decreased from 70.0% to 54.8%, Rottlerin could inhibite Mtb-Ag down regulation CD62L level of γδT cells (63.1%). Conclusion The CD62L expression of γδT cells could be ingibited partly by the inhibitor of PKCθ signal pathway may regulate L-selectin (CD62L) expression of activated human γδT cells.
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Objective The person's Mesangial Proliferative Glomemlonephritis (MsPGN) were divided into 3 types based on clini- cal manifestations: nephroticsyndrome, hematuria and proteinuria. To investigate the expression of CD11a, CD11b, CD62L and its signifi- cance in the kidneys patients with MsPGN. Methods CD11a, CD11b and CD62L expression in blood of MsPGN patients (n=35) were investigated by flow eytometry method, and the changes of these proteins in kidney were surveyed. Results In MsPGN, CD11a and CD11b expression in blood were significandy lower and CD62L expression in blood were markedly higher than that in normal humans. And the renal glomeruli of MsPGN also expressed CD11a, CD11b and CD62L. Conclusion The expression of CD11a, CD11b and CD62L are abnormal in MsPGN, and apoptosis may play certain role in the pathogenesis of MsPGN.
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Objective To construct the expression vector and express the recombinant human L-selectin in prokaryotic system, and purify the aimed protein for the preparation of rabbit anti-human L-selectin polyclonal antibody. Methods The partial cDNA of human L-selectin was amplified from the open reading frame (ORF) of N-terminus sequence of L-selectin containing 153 amino acids by PCR, then cloned into the prokaryotic expression vector pQE40 at restriction sites BamH Ⅰ and Hind Ⅲ. The recombinant expression plasmid pQE40-L-selectin was transformed into E. coli M15 for expression. The fusion protein including 6-His-tag was purified by Ni-NTA chromatographic column and analysed by SDS-PAGE. The purified protein was used to immune rabbit for preparing polyclonal antibody. Western blotting analysis and dot immunoblot assay (DIBA) were used to test the titer of the antiserum. Results The expression plasmid pQE40-L-selectin was constructed and confirmed with restriction enzyme digestion. The quantity of the purified protein from E. coli M15 was 600 mg ? L-1. By immuning the rabbit, the polyclonal antibody was successfully prepared. The results of dot immunoblot assay (DIBA) showed that the antiserum had the high titer (1 : 1 000). Conclusion The recombinant human L-seleclin prolein can express with high efficiency in E. coli M15. The prepared polyclonal antibody has a high titer.
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Objective To observe the changes of soluble L-selectin(sL-selectin) concentrations in children with viral encephalitis(VE) and probe its significance in pathogenesy and clinic.Methods Selecting 30 children with VE,including 14 mild case and 16 severe case.Collecting their venous blood and cerebrospinal fluid(CSF) in acute and recovery stage.Using double antibody sandwich enzyme-linked immunosorbent assay(ELISA) method to measure sL-selectin concentrations of serum and CSF and comparing with control group,respectively.Results SL-selectin concentrations from serum and CSF in the acute stage were significantly higher than those of control group(all P0.05).Conclusions L-selectin may participate in the pathologic course in VE.The concentrations of sL-selectin from serum and CSF are related with the patients′ condition and it has important reference value in judging patients′ condition,assessing their prognose and guiding clinical treatment.
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Objective To study the effect of the ECV-304 cells modified with LIF gene on the ex vivo culture of HSC/HPC in cord blood.Methods The ECV-304 cells were infected by Eukaryotic Expression plasmid pcDNA3.0LIF,and the positive ECV-304 cells were obtained by selected with G418.These cells were used to co-culture with CD34+ cells of cord blood.The phenotype of CD34+ and CD34+ CD54+、CD34+ CD62L+ primitive progenitors was detected by flow cytometry.Results The LIF gene can express in ECV-304 cells steadily.ECV-304 cells modified with LIF gene can improve expansion of CB CD34+、CD34+CD54+ and CD34+ CD62L+ cells while sustaining the expression of homing-related adhesion molecule.Conclusion The ECV-304 cells modified with LIF gene can not only significantly expand CB hematopoietic progenitor cells ex vivo,but the expanded CD34+ cells may well retain their homing ability.
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Objective: To study the expression of L selectin and its possible role in pulmonary injury in rat models undergoing cardiopulmonary bypass (CPB) and to investigate the protecive effect of fucoidin during the procedure. Methods: Models of CPB and pulmonary perfusion in rats were used to investigate the expression of L selectin pre and post operation. Its expression in antagonist group was also studied. Other indexes, including the concentration of SOD, MDA and MPO in rat lung tissues as well as PaO 2/FiO 2 and histological changes, were studied to illustrate the possible mechanism of L selectin in lung reperfusion injury. Results: The expression of L selectin increased after creating CPB but decreased after pulmonary infusion with fucoidin during the procedure, and lung injury relieved as well. Conclusion: L selectin might lead to lung injury during CPB, blocking its expression may relieve lung injury and enhance the recovery of lung function.
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Objective L-selectin is an important adhesion molecule,which is the lymph cell's homing receptor specifically expressed on leukocytes.A non-lymphoma cell line-HCa-F has the high potential ability of metastasis to peripheral lymph nodes,whose character is similar to lymphoma's.We want to explore whether the HCa-F cell's metastasis mechanism is also similar to lymphoma's,that is to say,whether HCa-F cell can also express L-selectin and the expression of L-selectin bring about the lymph node metastasis character. Methods First,?-fetoprotein,CD3 and CD20's monoclonal antibodies were used to identify the origin of HCa-F cell;then we analysed whether the HCa-F cell can express L-selectin by means of flow cytometry analysis,RT-PCR,and Western blotting;finally,Stamper-Woodruff method was used to examine the L-selectin's function. Results Immunostaining showed that the HCa-F cell was ?-fetoprotein positive,CD3 and CD20 negative cell line,which suggests that it is a liver cell line.And results of flow cytometry,RT-PCR,and Western blotting showd that HCa-F cell can express L-selectin.Additionally,anti-L-selectin's antibody can inhibit the adhesion between HCa-F cells and frozen section of mouse lymph node.Conclusion L-selectin expressed on HCa-F cells is a functional molecule which can facilitate HCa-F cell's metastasis to lymph nodes.
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Objective To investigate the effects of 2-methoxyl estradiol(2-ME)on the expression of L-selectin and its mRNA and the relationship between L-selectin and bcr/abl fusion gene expression in chronic myelocytic leukemia(CML)K562 cells.Methods The cultured K562 cells were divided into three groups.The control group,K562 cells were cultured without 2-ME treatment.The experimental group,K562 cells were cultured in the medium containing different concentrations of 2-ME(1,2,4,8 and 16?mol/L)for 36h.The negative control group,K562 cells were replaced by water without RNase in the medium containing different concentrations of 2-ME for 36h.The expression of L-selectin protein in K562 cells was determined by flow cytometry(FCM)method.The expression of L-selectin mRNA in K562 cells was detected by half-quantitative reverse transcription-polymerase chain reaction(RT-PCR).The bcr/abl fusion gene expression in K562 cells was detected by quantitative SYBR Green Ⅰ real-time RT-PCR.Results After being treated with 2-ME at different concentrations for 36 hours,the expression of L-selectin protein and its mRNA increased in K562 cells in a concentration-dependent manner,and the expression level in the experimental group was higher than that in the control group(P
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AIM:To explore the changes of L-selectin expression and F-actin distribution of leukocytes from pancreatic circulation in the experimental acute necrotizing pancreatitis (ANP) rats and impact of Chinese herbs west China pancreatitis yi hao (WPY) on them in vivo. METHODS:Forty-four Wistar rats were randomized into control group,ANP group and ANP WPY treated group. The leukocytes from blood of splenic vein were incubated with PE-anti-CD62L for L-selectin and TRTIC-Phalloidin for F-actin,then the ratio of CD62L positive granulocytes was measured by flow cytometry and leukocytes were taken images with confocal laser scanning microscope. The distribution of F-actin at the border or central part of leukocyte was analyzed by using Mias system. RESULTS:The ratio of CD62L positive granulocytes increased in ANP rats,but those increases at 6 h (31.78%?12.37%) and 12 h (29.83%?13.73%) were significant [vs control group(11.96%?5.32%),P
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AIM: To study the effect of selectin on lung injury in rabbits with diabetic ketoacidosis(DKA). METHODS: 12 male New Zealand rabbits were divided into experimental group (DK groups, intravenous injection of streptozocin and alloxan monohydroate, n =6) and normal control group (received same dose of saline, n =6). After 72 hours, all the animals were killed and blood and lung tissue were collected. Blood gases, expressions of selectin-P and selectin-L were detected. RESULTS: The expression of selectin in DK group were higher than that in NS group. CONCLUSION: These results indicate that selectin expression may play an important role in lung injury during diabetic ketoacidosis.
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BACKGROUND: The CD4+ T-helper cells comprise functionally distinct subsets of Th1 and Th2 cells that are distinguished on the basis of differential cytokines production. Th1 cells secrete interferon-gamma, lymphotoxin, interleukin-2. Th2 cells produce interleukin-4, interleukin-5, interleukin-10. A previous study shown that Th2 cells and their cytokines increased in patients with atopic asthma. We compared cytokines(IL-4, IFN-gamma) activity and subpopulation of T-lymphocytes in peripheral blood from atopic asthmatics versus non-asthmatics. METHOD: Fifteen patients with atopic asthma(nine men, six women), twelve patients with chronic bronchitis (six men, six women), five healthy persons(three men, two women) were studied. Activity of IL-4, IFN-gamma and T-cell subpopulation in peripheral blood were estimated. RESULTS: Patients had a median age of 55yr. The mean activity of IL-4 of asthmatics was significantly increased(control 0.75+/-1.1pmol/L, atopic asthmatics 3.50+/-0.75pmol/L, chronic bronchitis 2.01+/-1.2pmol/L), but IFN- was not significantly increased. In the T lymphocyte subsets the percent of CD62L+ T-lymphocytes of asthmatics was not significantly increased(control 16.7+/-16.4 %, atopic asthmatics 24.8+/-23.6 %, chronic bronchitis 17.0+/-16.9%). CONCLUSION: In this study elevated production of IL-4 was observed in atopic asthmatics. CD62L+ T-lymphocytes was not increased in atopic asthma.
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Humans , Male , Asthma , Bronchitis, Chronic , Cytokines , Interferon-gamma , Interleukin-10 , Interleukin-2 , Interleukin-4 , Interleukin-5 , Lymphotoxin-alpha , T-Lymphocyte Subsets , T-Lymphocytes , Th1 Cells , Th2 CellsABSTRACT
AIM: To explore the changes of L-selectin expression on peripheral blood polymorphonuclear leukocytes (PMNs) and their significance in rats with acute lung injury (ALI). METHODS: ALI model in rat was established by intravonous injection of E. coli endotoxin (ET). The expression of L-selectin on peripheral blood PMNs was measured by immunofluorescence and flow cytometry.RESULTS: The contribution of L-selectin fluorescence signal was on the surface of PMNs membrane. The expression of L-selectin on poeripheral blood PMN was significantly lower at 5 min after injection of ET and the lowest during 15 min to 30 min, then gradually increased, but the expression of L-selectin on PMN was lower at 60 min after injection of ET than that of control animal.CONCLUSION: In physiological state, L-selectin were expressed on the surface of PMN membrane. The protein expression of L-selectin on PMNs reduced rapidly after injection of ET and the lowest at 15 min, then gradually increased. L-selectin may play a role in the development of ALI.
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Objective To explore the association between L-selectin gene P213S polymorphism and angina pectoris.Methods L-selectin gene P213S polymorphism in 138 patients with angina pectoris and 156 controls was detected by polymerase chain reaction restriction fragment length polymorphism(PCR-RFLP).The relationship between gene polymorphism of L-selectin and levels of serum lipids were also studied.Results L-selectin genotype frequencies of PP,PS,SS were 60.1%,36.3%,3.6% and 44.9%,48.1%,7.0% in angina pectoris group and control group respectively.Allele frequencies of P,S were 78.3%,21.7% and 68.9%,31.1% in angina pectoris group and control group respectively.There was significant differences of frequencies of genotype and allele of L-selectin P213S polymorphism between angina pectoris group and control group(P