Platelet transfusion in patients with liver cirrhosis crosstalk with neutrophil: Prospective study

ABSTRACT Introduction: Immune dysfunction and thrombocytopenia are common features in liver cirrhosis. Platelet transfusion is the most widely used therapeutic approach for thrombocytopenia when indicated. The transfused platelets are prone to lesions during their storage that empower their interaction with the recipient leucocyte. These interactions modulate the host immune response. The impact of platelet transfusion on the immune system in cirrhotic patients is little understood. Therefore, this study aims to investigate the impact of platelet transfusion on neutrophil function in cirrhotic patients. Methods: This prospective cohort study was implemented on 30 cirrhotic patients receiving platelet transfusion and 30 healthy individuals as a control group. EDTA blood samples were collected from cirrhotic patients before and after an elective platelet transfusion. Flowcytometric analysis of neutrophil functions (CD11b expression and PCN formation) was performed. Results: There was a significant increase in expression of CD11b on neutrophils and Frequency of platelet-complexed neutrophils (PCN) in patients with cirrhosis compared with controls. Platelet transfusion increased level of CD11b and the frequency of PCN even more. There was a significant positive correlation between change in PCN Frequency pefore and after transfusion and the change in expression of CDllb among cirrhotic patients. Conclusions: Elective platelet transfusion appears to increase level of PCN in cirrhotic patients, moreover, exacerbate the expression of activation marker CDllb on both neutrophils and PCN. More research and studies are needed to corroborate our preliminary findings.

The role of neutrophil CD 11b, soluble CD 14 subtype and mitochondrial coupling factor-6 in the risk stratification of disease outcome in neonatal sepsis and its clinical significance / 中国医师进修杂志

Objective:To investigate the role of neutrophil CD 11b (nCD 11b), soluble CD 14 subtype (sCD 14-St) and mitochondrial coupling factor-6 (CF-6) in the risk stratification of disease outcome in neonatal sepsis and its clinical significance. Methods:The clinical data of 121 septic neonates from July 2019 to March 2020 in Shanxi Children′s Hospital were retrospectively analyzed. According to the neonatal critical illness score (NCIS), the neonates were divided into non-critical group (NCIS>90 scores) with 35 cases, critical group (NCIS 70 to 90 scores) with 49 cases, very critical group (NCIS<70 scores) with 37 cases. There were 25 cases with poor prognosis (death), and 96 cases with good prognosis (survival). The C-reactive protein (CRP), procalcitonin (PCT), nCD 11b, sCD 14-St and CF-6 before treatment were detected. The correlation between nCD 11b, sCD 14-St, CF-6 and disease severity was analyzed by Spearman method; the value of nCD 11b, sCD 14-St and CF-6 in predicting poor disease outcome in sepsis neonates was analyzed by the receiver operating characteristic (ROC) curve. Results:The nCD 11b, sCD 14-St, CF-6, PCT and CRP in critical group and very critical group were significantly higher than those in non-critical group: (414.68 ± 93.29) and (532.74 ± 101.85) MFI vs. (325.45 ± 71.90) MFI, (892.40 ± 113.72) and (1 249.53 ± 95.41) ng/L vs. (784.66 ± 103.72) ng/L, (84.79 ± 28.35) and (121.66 ± 34.27) ng/L vs. (42.59 ± 13.51) ng/L, (19.24 ± 6.30) and (34.96 ± 11.95) μg/L vs. (8.89 ± 2.24) μg/L, (109.49 ± 36.77) and (247.13 ± 82.06) mg/L vs. (56.84 ± 17.25) mg/L; the indexes in very critical group were significantly higher than those in critical group, and there were statistical differences ( P<0.05). Spearman correlation analysis result showed that nCD 11b, sCD 14-St and CF-6 were positively correlated with disease severity in sepsis neonates ( r = 0.719, 0.813 and 0.823; P<0.01). The nCD 11b, sCD 14-St, CF-6, PCT and CRP in poor prognosis neonates were significantly higher than those in good prognosis neonates: (618.58 ± 146.92) MFI vs. (374.55 ± 120.03) MFI, (1 516.91 ± 194.38) ng/L vs. (828.13 ± 175.67) ng/L, (165.84 ± 25.63) ng/L vs. (62.51 ± 16.75) ng/L, (43.46 ± 10.14) μg/L vs. (20.19 ± 6.30) μg/L and (321.09 ± 94.56) mg/L vs. (88.24 ± 29.19) mg/L, and there were statistical differences ( P<0.01). ROC curve analysis result showed that the area under the curve (AUC) of nCD 11b, sCD 14-St and CF-6 for predicting poor disease outcome in sepsis neonates were 0.763, 0.796 and 0.838 (95% CI 0.678 to 0.836, 0.713 to 0.864 and 0.760 to 0.899), and the AUC of combination the 2 indexes was 0.921 (95% CI 0.858 to 0.962). Conclusions:The nCD 11b, sCD 14-St and CF-6 are associated with the disease severity and prognosis in sepsis neonates, and can be used as markers for risk stratification of disease outcome and assessment prognosis.

Mechanism of lung adenocarcinoma-infiltrating CD8+T cells expressing CD39 / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨肿瘤浸润CD8+ T细胞表达CD39的可能机制。方法：通过TCGA数据库的肺腺癌（LUAD）组织及正常肺组织转录组数据分析CD39在LUAD组织和正常肺组织中的表达差异及其对患者预后的影响，分析CD39表达与T细胞浸润、激活的关系。用小鼠LUAD Lewis细胞建立小鼠皮下移植瘤模型，FCM检测淋巴结、脾脏以及移植瘤组织中CD8+ CD39+ T细胞。收集Lewis细胞培养上清液作为条件培养基（CCM），免疫磁珠法（MACS）分选CD8+ T细胞、CD11b+细胞；在培养基中分别加入CCM、IL-6和采取非接触或接触培养方式进行培养，探索CD8+ T细胞表达CD39的可能机制。结果：CD39在LUAD组织中呈低表达（P<0.01），其表达水平与LUAD患者OS、T细胞浸润和激活水平均呈正相关（P<0.05或P<0.001）。FCM检测结果显示，在移植瘤组织中CD8+CD39+ T细胞的比例明显高于淋巴结及脾脏（P<0.01）；CCM及IL-6不能直接诱导CD8+ T细胞表达CD39，非接触共培养CD11b+细胞与CD8+T细胞也不能诱导CD8+ T细胞表达CD39，CD11b+细胞与CD8+ T细胞直接接触共培养可诱导CD8+ T细胞表达CD39（P<0.01）。结论：CD11b+细胞通过直接接触方式诱导肿瘤浸润CD8+ T细胞表达CD39，CD39可能是LUAD特异性CD8+ T细胞的分子标志。

Preparation of 68Ga labeled pre-targeted molecular probe based on bio-orthogonal Diels-Alder click chemistry and CD11b receptor targeted imaging in CT26 colon cancer mouse models / 中华核医学与分子影像杂志

Objective:To prepare a 68Ga labeled probe targeting integrin alpha M(CD11b) receptor, namely 68Ga-1, 4, 7-triazacyclononane-1, 4, 7-triacetic acid-Glycine-Arginine-Glutamate-Arginine-Glutamate-polyethylene glycol 11-1, 2, 4, 5-terazine/CD11b antibody-F(ab′) 2-trans-cyclooctene ( 68Ga-NOTA-Polypeptide-PEG 11-Tz/anti-CD11b-F(ab′) 2-TCO), and to explore its feasibility as a molecular probe for CD11b receptor through microPET imaging. Methods:Immunofluorescence was used to detect the expression of CD11b on the surface of RAW264.7 cell. CD11b specific monoclonal antibody (M1/70) was conjugated with TCO, and anti-CD11b-F(ab′) 2-TCO fragment was obtained. The ligand NOTA-Polypeptide-PEG 11-Tz was labeled with 68Ga, and its specific activity and radiochemical purity were detected. Pre-targeted cell binding experiment was conducted to evaluate the binding ability of molecular probe. CT26 colon cancer bearing mouse models were established, and then pre-targeted biodistribution and imaging experiments were performed. Immunohistochemical experiment was used to verify the expression of CD11b receptor in tumor. The one-way analysis of variance was used to compare the data. Results:The results of immunofluorescence demonstrated CD11b receptor was highly expressed on the surface of RAW264.7 cell. Anti-CD11b-F(ab′) 2-TCO fragment was verified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). 68Ga-NOTA-Polypeptide-PEG 11-Tz was successfully synthesized, with the labeling efficiency of 94.6%. The specific activity was 7.0-7.4 MBq/μg, and the radiochemical purity was higher than 95%. Pre-targeted cell binding experiment confirmed that the molecular probe bound to the CD11b receptor. The biodistribution and imaging experiments showed that the kidney radioactivity uptake was high at pre-targeted 4, 12 and 24 h intervals, which proved that probe was excreted through the urinary system. In addition, molecular probe had higher radioactive uptake at the tumor site, with the tumor/muscle ratios of 9.23±1.45, 12.53±1.36 and 10.74±1.11 ( F=848.8, P<0.05). When the radioligand was injected 1 h after the pre-positioned 12 h interval, the images contrast was the best, with the standardized uptake value (SUV) in tumor and muscle of 0.67±0.12, 0.09±0.04, respectively. Immunohistochemistry verified the highly expression of CD11b receptor in tumor. Conclusions:The pre-targeted molecular probe 68Ga-NOTA-Polypeptide-PEG 11-Tz/anti-CD11b-F(ab′) 2-TCO is successfully synthesized. The molecular probe has targeting ability for CD11b + colon cancer, and is expected to be used as a tracer targeting CD11b receptor in vivo.

Inhibition of GSK-3 by Tideglusib Suppresses Activated Macrophages and Inflammatory Responses in Lipopolysaccharide-stimulated RAW 264.7 Cell Line

@#Introduction: Glycogen synthase kinase-3 (GSK-3) is an important immune regulator that controls inflammation via inhibition of its protein kinase activities. Persistent inflammatory responses through the activation of immune cells and excessive production of immune mediators may cause tissue destruction and implicated in the development of chronic inflammatory diseases. The objective of this study was to examine the role of Tideglusib, a GSK-3 inhibitor, in inflammatory responses elicited through macrophage activation by investigating the expression of cell surface biomarkers and inflammatory molecule levels. Method: The effects of GSK-3 inhibition by Tideglusib on the expression of CD11b and CD40 and secretion of pro-inflammatory cytokines in the lipopolysaccharide (LPS)-activated macrophage-derived RAW 264.7 cells were determined by flow cytometry, while the presence of nitric oxide (NO) was determined by Griess assay. Results: Stimulation of RAW 264.7 cells with LPS increased substantial levels of CD11b and CD40 expressions, and secretion of NO, TNF-α, and MCP-1. However, the expression of these molecules was suppressed through inhibition of GSK-3. Conclusion: These findings suggest the significant role of Tideglusib to limit the upregulation of immune responses in activated macrophages, and as a potential anti-inflammatory drug for the intervention and treatment of inflammatory diseases.

Effect of curcumin on expressions of CD11b and CD19 in peripheral blood of heat stroke rats in a simulation dry-heat environment / 中华危重病急救医学

Objective? To?investigate?the?effects?of?different?doses?of?curcumin?on?the?levels?of?immune?factors?CD11b?and?CD19?in?peripheral?blood?of?heat?stroke?rats?in?a?simulation?dry-heat?environment.? Methods? 160?SPF??healthy?male?Sprague-Dawley?(SD)?rats?were?selected?and?divided?into?different?groups?according?to?random?number?table?method:?normal?saline?(NS)?control?group?(given?NS),?solvent?control?group?[given?sodium?carboxymethylcellulose?(CMCNa)],?and?curcumin?low,?medium?and?high?dose?pretreatment?group?(given?0.05,?0.10,?0.20?mg/g?of?curcumin+0.5%?CMCNa?solution).?There?were?32?rats?in?each?group,?and?were?challenged?only?by?10?mL·kg-1·d-1?lavage,?and?continuous?dosing?for?7?days.?On?the?8th?day,?rats?were?challenged?at?ambient?temperature?(41.0±0.5)?℃,?relative?humidity?(10±1)%?of?the?northwest?in?the?special?environment?of?artificial?lab,?placed?in?0?(normal?temperature),?50,?100?and??150?minutes?respectively.?The?levels?of?CD19?and?CD11b?in?peripheral?blood?of?each?rat?were?detected?by?flow?cytometry?instrument.? Results? With?the?extension?of?time?in?the?simulated?dry?and?heat?environment,?the?level?of?CD11b?in?peripheral?blood?was?gradually?increased?in?each?group,?and?the?peak?value?was?reached?at?150?minutes,?the?NS?control?group,?solvent?control?group?and?curcumin?low,?medium?and?high?dose?pretreatment?groups?were?0.346±0.013,?0.342±0.013,?0.342±0.012,?0.325±0.012,?and?0.281±0.012,?respectively.?In?each?group,?the?level?of?CD19?was? first?increased?and?then?decreased,?reaching?its?peak?value?at?100?minutes,?and?the?level?of?the?NS?control?group,?solvent?control?group?and?curcumin?low,?medium?and?high?dose?pretreatment?groups?were?0.586±0.010,?0.601±0.014,?0.684±0.009,?0.613±0.012?and?0.604±0.006,?respectively.?The?level?of?CD11b?in?the?curcumin?medium?and?high?dose?pretreatment?groups?were?significantly?lower?than?those?in?the?NS?control?group?and?solvent?control?group??(50?minutes:?0.237±0.011,?0.188±0.006?vs.?0.283±0.009,?0.289±0.012;?100?minutes:?0.260±0.010,?0.248±0.008?vs.?0.293±0.008,?0.290±0.007,?all?P?<?0.05),?and?after?placement?for?150?minutes,?the?level?of?CD11b?in?the?curcumin?high?dose?pretreatment?group?was?significantly?lower?than?that?in?the?NS?control?group,?solvent?control?group?and?curcumin?low?dose?pretreatment?group?(0.281±0.012?vs.?0.346±0.013,?0.342±0.013,?0.342±0.012,?all?P?<?0.05).?The?level?of?CD19?in?the?curcumin?low,?medium?and?high?dose?pretreatment?groups?were?significantly?higher?than?those?in?the?NS?control?group?and?solvent?control?group?at?50?minutes?in?the?dry?and?hot?environment?(0.394±0.001,?0.436±0.009,?0.553±0.011?vs.?0.205±0.005,?0.197±0.003,?all?P?<?0.05),?at?100?minutes,?the?level?of?CD19?in?the?curcumin?low?dose?pretreatment?group?was?significantly?higher?than?that?in?the?NS?control?group?and?solvent?control?group?(0.684±0.009?vs.?0.586±0.010,?0.601±0.014,?both?P?<?0.05),?there?was?no?significant?difference?in?CD19?level?between?the?other?dose?pretreatment?groups?and?NS?control?group;?at?150?minutes,?there?was?no?significant?difference?in?CD19?level?between?the?curcumin?pretreatment?groups,?the?NS?control?group,?and?the?solvent?control?group.?The?peripheral?blood?immune?factors?CD11b?and?CD19?levels?in?the?NS?control?group?and?solvent?control?group?were?not?significantly?changed,?and?there?was?no?significant?difference?between?two?groups.? Conclusion? Curcumin?pretreatment?can?reduce?the?level?of?CD11b?and?increase?the?level?of?CD19?in?peripheral?blood?of?rats?with?dry?heat?stroke?in?the?early?and?middle?stages,?which?may?enhance?the?heat?resistance?and?prevent?the?occurrence?of?multiple?organ?dysfunction?by?increasing?the?body?immunity,?and?this?effect?has?nothing?to?do?with?the?dose?of?curcumin.

PMN-CD11b Levels and Its Correlation with Left Atrial Diameter in Patients with Atrial Fibrillation / 中山大学学报(医学科学版)

@#【Objective】To observe the levels of CD11b expressions in the surface of neutrophils in serum，and its correlation with left atrial diameter ，and to study the inflammatory mechanisms in atrial fibrillation（AF） patients. 【Methods】Clinical characteristics and blood samples of AF group and sinal rhythm group were collected. CD11b levels in the surface of neutrophils were examined by flow cytometry. Left atrial diameter was examined by echocardiography. 【Results】The AF group included 85 patients ：36 with paroxysmal AF；26 with persistent AF；23 with permanent AF. The sinal rhythm group includes 57 patients. PMN- CD11b levels were significantly higher in Af group than in sinal rhythm group（P＜0.01）. The left atrial diameter was larger in AF group than in sinal rhythm group（P＜0.01）. Multivariate analysis revealed that PMN-CD11b，left atrial diameter and Valvular heart disease were independent predictors of atrial fibrillation.【Conclusions】PMN- CD11b levels were elevated in paroxysmal AF when AF was present and in persistent， permanent AF patients，implying atrial fibrillation was closely related to inflammation；PMN-CD11b were correlated with left atrial diameter，inflammation might participate in the atrial structural remodeling in AF patients ；PMN-CD11b levels were elevated in AF patients with high risk thrombosis，inflammation might have some value to embolic risk stratification according to CHA2DS2-VASc score.

Expressions of cell activating factors in patients with malignant lymphoma and their clinical significances / 白血病·淋巴瘤

Objective To investigate the effects of endothelial cell, neutrophil and platelet activation on the development and thrombophilia of malignant lymphoma. Methods The levels of cell activating factors soluble thrombomodulin (sTM) in 40 patients with malignant lymphoma and 30 healthy controls in Jiangdu People ''s Hospital of Yangzhou from October 2009 to November 2016 were tested by enzyme linked immunoabsorbent assay (ELISA). The flow cytometry (FCM) method was used to measure the level of CD11b on the surface of anti-freezing whole blood neutrophil and CD62p on the surface of platelet. The results were analyzed by ANOVA and LSD-t test. Results The level of sTM in patients with malignant lymphoma before treatment was significantly higher than that after treatment and in the control group [(49.7±9.3) ng/ml vs. (6.3± 1.8) ng/ml, (5.6±1.5) ng/ml respectively;F=736.633, P=0.000]. There were significant differences between the levels before and after treatment and before treatment and in the control group (t= 33.789, P= 0.000;t=31.782, P=0.000). The average fluorescence intensity (MFI) of neutrophil surface CD11b in patients with malignant lymphoma before treatment was significantly higher than that after treatment and in the control group (184 ± 43 vs. 118 ± 12, 112 ± 15 respectively; F=101.845, P=0.000). There were significant differences between the levels before and after treatment and before treatment and in the control group (t=12.228, P= 0.000; t= 12.184, P= 0.000). The level of platelet surface CD62p in patients with malignant lymphoma before treatment was significantly higher than that after treatment and in the control group [(6.4 ± 2.4) ％ vs. (1.2 ± 0.7) ％, (1.3 ± 0.5) ％ respectively; F= 141.481, P= 0.000]. There were significant differences between the levels before and after treatment and before treatment and in the control group (t=15.010, P= 0.000; t= 13.679, P= 0.000). Conclusions The levels of cell activating factors might be correlated with the progression of malignant lymphoma. Furthermore, endothelial cell, neutrophil and platelet activation have accelerated the development of malignant lymphoma, and the interaction among them may result in the prethrombotic state.

Changes in natural killer cell subsets defined by CD11b and CD27 in spleen of septic mice / 中华麻醉学杂志

Objective To evaluate the changes in natural Killer (NK) cell subsets definetd by CD11b and CD27 in the spleen of septic mice.Methods A total of 168 pathogen-free healthy male C57BL/6 mice,weighing 20-30 g,aged 8-10 weeks,were divided into 2 groups (n=84 cach) using a random number table:sham operation group (Sham group) and sepsis group (Sep group).Sepsis was induced by cecal ligation and puncture in chloral hydrate-anesthetized mice.Thirty mice in each group were selected to assess the survival within 4 days after operation,and the survival rate was calculated.At 2,4,6,24,48 and 72 h after operation (T1-6),6 mice were randomly selected,and blood samples were taken from the eyeballs for determination of serum tumor necrosis factor-alpha and interleukin-10 concentrations by enzyme-linked immunosorbent assay.Six mice were randomly selected at T4-6 and sacrificed,and the spleens were removed for measurement of the percentage of NK cells and their subsets by flow cytometry.Results Compared with Sham group,the survival rate was significantly decreased at different time points,the serum necrosis factor-alpha concentration at T1-3,6 and serum interleukin-10 concentration at T3-6 were increased,the percentage of NK cells was decreased at T4-6,the percentage of CD27-CD11b+NK cells was decreased and the percentage of CD27+CD11b+ and CD27+CD11b-NK cells was increased at T5,the percentage of CD27-CD11b-NK ceils was decreased at T4,5,and the percentage of CD27-CD11b-NK cells was increased at T6 in Sep group (P＜0.05).Conclusion Changes in NK cell subsets defined by CD11b and CD27 in the spleen play an important role in the development of sepsis in mice.

Identification and establishment of sorting method for isolating CD11b+ myeloid cells in human hepatic metastases from colorectal cancer / 中国病理生理杂志

AIM:To establish a method for obtaining specific cells in solid tumor tissue by sorting of CD11b + myeloid cells in hepatic metastases from colorectal cancer.METHODS:Tumor tissues were prepared into single cell suspension by mechanical method combined with enzyme digestion,and then the CD11 b + myeloid cells were isolated by flow cytometry.The sorted cells were identified by immunocytochemistry.The viability and morphologiy of the sorted cells were evaluated by Giemsa and Typan blue staining.The cell purity was evaluated by flow cytometry.RESULTS:Sufficient numbers of CD11b+cells with high purity were isolated by sorting with flow cytometry from the single cell suspension prepared by mechanical and enzyme digestion.The purity of the cells was confirmed by statistical analysis (P ＜ 0.05).The positive rates of the cells before and after sorting were significantly different (P ＜0.01).The positive cells were verified by immunocytochemical method.Meanwhile,the sorted cells had complete morphology and good activity.CONCLUSION:The CD11b + myeloid cells in solid tumor tissue can be isolated by flow cytometry from the machine-enzyme digestion suspension with high purity,good activity and complete morphology.

Identification and establishment of sorting method for isolating CD11b+ myeloid cells in human hepatic metastases from colorectal cancer / 中国病理生理杂志

AIM:To establish a method for obtaining specific cells in solid tumor tissue by sorting of CD11b + myeloid cells in hepatic metastases from colorectal cancer.METHODS:Tumor tissues were prepared into single cell suspension by mechanical method combined with enzyme digestion,and then the CD11 b + myeloid cells were isolated by flow cytometry.The sorted cells were identified by immunocytochemistry.The viability and morphologiy of the sorted cells were evaluated by Giemsa and Typan blue staining.The cell purity was evaluated by flow cytometry.RESULTS:Sufficient numbers of CD11b+cells with high purity were isolated by sorting with flow cytometry from the single cell suspension prepared by mechanical and enzyme digestion.The purity of the cells was confirmed by statistical analysis (P ＜ 0.05).The positive rates of the cells before and after sorting were significantly different (P ＜0.01).The positive cells were verified by immunocytochemical method.Meanwhile,the sorted cells had complete morphology and good activity.CONCLUSION:The CD11b + myeloid cells in solid tumor tissue can be isolated by flow cytometry from the machine-enzyme digestion suspension with high purity,good activity and complete morphology.

Exacerbation of Japanese Encephalitis by CD11c(hi) Dendritic Cell Ablation Is Associated with an Imbalance in Regulatory Foxp3⁺ and IL-17⁺CD4⁺ Th17 Cells and in Ly-6C(hi) and Ly-6C(lo) Monocytes

Japanese encephalitis (JE) is neuroinflammation characterized by uncontrolled infiltration of peripheral leukocytes into the central nervous system (CNS). We previously demonstrated exacerbation of JE following CD11c(hi) dendritic cell (DC) ablation in CD11c-DTR transgenic mice. Moreover, CD11c(hi) DC ablation led to abnormal differentiation of CD11b⁺Ly-6C(hi) monocytes and enhanced permeability of the blood-brain barrier (BBB), resulting in promoting the progression of JE. Here, we examined changes in lymphoid and myeloid-derived leukocyte subpopulations associated with pro- and anti-inflammation during JE progression. The analyses of this study focused on regulatory CD4⁺Foxp3⁺ regulatory T cells (Tregs), IL-17⁺CD4⁺ Th17 cells, and CD11b⁺Ly-6C(hi) and Ly-6C(lo) monocytes. CD11c(hi) DC ablation resulted in the accumulation of IL-17⁺CD4⁺ Th17 cells in the CNS, thereby leading to lower ratio of Tregs to Th17 cells. This result was corroborated by the higher expression levels of IL-17 and RORγT in CD4⁺ T cells from the brains of CD11c(hi) DC-ablated mice. In addition, CD11c(hi) DC-ablated mice showed higher frequency and total number of inflammatory CD11b⁺Ly-6C(hi) monocytes, whereas CD11b⁺Ly-6C(lo) monocytes were detected with lower frequency and total number in CD11c(hi) DC-ablated mice. Furthermore, CD11c(hi) DC ablation altered the phenotype and function of CD11b⁺Ly-6C(lo) monocytes, resulting in lower levels of activation marker and anti-inflammatory cytokine (IL-10 and TGF-β) expression. Collectively, these results indicate that CD11c(hi) DC ablation caused an imbalance in CD4⁺ Th17/Treg cells and CD11b⁺Ly-6C(hi)/Ly-6C(lo) monocytes in the lymphoid tissue and CNS during JE progression. This imbalanced orchestration of pro- and anti-inflammatory leukocytes following CD11c(hi) DC ablation may contribute to the exacerbation of JE.

Effects of Ginkgo biloba tablet on MPA, CD11 b and MCP-1 levels in patients with unstable angina pectoris / 中国临床药理学与治疗学

AIM:To observe the levels of monocyte-platelet aggregates (MPA) and markers of activated monocytes in patients with unstable angina pectoris (UAP) accepting Ginkgo biloba tablet treatments,and to explore its mechanisms for cardiovascular disease treatments.METHODS:The levels of MPA,CD11b,and MCP-1 were measured in 92 unstable angina pectoris (UAP) and 42 stable angina pectoris (SAP).The UAP patients were randomly assigned into routine treatment group (control group) and combined tablet treatment group (Ginkgo biloba group).The efficacy was assessed,and the levels of MPA,CD11b,and MCP-1 were measured after 28 days of treatment,respectively.RESULTS:The levels of MPA,CD11b,and MCP-1 in UAP group were higher than those in SAP group (P＜0.001).The levels of MPA and CD11b were positively correlated with MCP-1 level (P ＜ 0.01).The total rate of effective Ginkgo biloba tablet treatment was higher than that of non-Ginkgo biloba tablet treatment (P ＜ 0.05).After 28 days of treatments,the levels of MPA,CD11b,and MCP-1 in Ginkgo biloba group were significantly lower than those in control group (P ＜ 0.001).In total effective treatment group,the levels of MPA,CD11b,and MCP-1 were significantly lower after treatment than those before treatment (P ＜ 0.001),and the decreased rates of these markers after treatment were also much higher (P ＜ 0.01).CONCLUSION:There is an obvious efficacy of Ginkgo biloba tablet on unstable angina pectoris by down-regulating the levels of MPA,CD11b and MCP-1.

Expression and significance of neutrophil surface adhesion molecules CD11b in type 2 diabetes mellitus patients with cytomegalovirus infection / 中国基层医药

Objective To study diagnosis significance of neutrophil surface adhesion molecules including CD11b,in type 2 diabetes mellitus patients with cytomegalovirus (CMV)infection.Methods Blood samples from 139 cases of type 2 diabetes mellitus patients were collected including 99 male and 40 female cases between Septem-ber 2011 and January 2015.Healthy male 20 cases and female 13 cases were collected as healthy control group from outpatients in the same period.Type 2 diabetes mellitus patients were divided into high active CMV infection group, CMV infection group and the negative control group,and healthy group as normal control group according to CMV -pp65 antigen detection.Neutrophil surface adhesion molecule CD11b was analyzed by flow cytometry and compared respectively.ROC curve of CD11b in 2 diabetes mellitus patients was made.Results The expression ratio of CD11b in high active CMV infection group and CMV infection group in type 2 diabetes mellitus patients was (41.25 ± 5.33)%.There was significant difference (P =0.019)compared with negative control group (61.13 ±5.28)% and there was significant difference (P =0.009)compared with negative control group (64.21 ±6.02)%;optimal cut -off value of CD11b in high active CMV infection group was 45.65%,the sensitivity was 100.00%,the specificity in high active CMV infection group was 60.12%,and the area under the receiver -operating curve(ROC)was 0.628;optimal cut -off value of CD11b in CMV infection group was 70.21%,the sensitivity of CD11b was 87.69%,the specificity of CD11b was 99.22%,and the area under of ROC of CD11b was 0.991.Conclusion The neutrophils CD11b expression in type 2 diabetes mellitus patients with CMV infection is down regulation.CD11b expression can be used as a laboratory diagnosis basis of CMV infection.

Effects of electroacupuncture on the activities of gliocytes of cervical spinal cord in rats with thyroid incisional pain / 中国针灸

<p><b>OBJECTIVE</b>To observe the impacts of electroacupuncture (EA) on microgliacytes and astrocytes of cervical spinal cord in rats with thyroid incisional pain and explore the mechanism of acupuncture anesthesia in thyroid surgery.</p><p><b>METHODS</b>Sixty Wistar male rats were randomized into a normal group, a model group, a Futu (LI 18) group, a Hegu (LI 4)-Neiguan (PC 6) group and a Zusanli (ST 36)-Yanglingquan (GB 34) group, 12 rats in each one. Except the normal group, a longitudinal incision, about 1.5 cm in length was done along the neck midline in the rats of the rest groups to prepare the model of thyroid incisional pain. In the Futu (LI 18) group, the Hegu (LI 4)-Neiguan (PC 6) group and the Zusanli (ST 36)-Yanglingquan (GB 34) group, after modeling for 4 h, 24 h and 48 h, EA was applied to bilateral "Futu" (LI 18), "Hegu" (LI 4) "Neiguan" (PC 6) and"Zusanli" (ST 36) "Yanglingquan" (GB 34) separately, once a day, continuously for 3 days. In the normal group and the model group, no any intervention was applied. The thermal radiant apparatus was used to detect the thermal pain threshold (PT). The fluorescence quantitative RT-PCR and the Western blotting (WB) were used to determine the expressions of protein and gene of microglia activation markers Iba1 and CD11b and the astrocyte specific protein marker, glial fibrillary acid protein (GFAP) in cervical spinal cord (Cto C) after intervention in the rats of each group.</p><p><b>RESULTS</b>After intervention, as compared with the normal group, in the model group, the neck PT was reduced apparently (<0.05), the expressions of Iba1 and CD11b and GFAP mRNA as well as the protein expressions in the spinal cord of Cto Cwere up-regulated apparently (<0.05,<0.01). As compared with the model group, in the Futu (LI 18) and the Hegu (LI 4)-Neiguan (PC 6) group, PT was increased significantly (both<0.05) and that did not change apparently in the Zusanli (ST 36)-Yanglingquan (GB 34) group (>0.05). In the Futu (LI 18) group, the protein and gene expressions of Iba1, CD11b and GFAP were lower than those in the model group (all<0.05). In the Hegu (LI 4)-Neiguan (PC 6) group, the expressions of Iba1 mRNA, CD11b protein, GFAP mRNA and protein were all lower apparently than those in the model group (all<0.05). In the Zusanli (ST 36)-Yanglingquan (GB 34) group, the expressions of Iba1, CD11b and GFAP proteins were not different significantly as compared with the model group (all>0.05). In the Zusanli (ST 36)-Yanglingquan (GB 34) group, the expressions of Iba1 mRNA and CD11b mRNA and protein expressions in the spinal cord of Cto Cwere higher apparently than those in the Futu (LI 18) group (<0.01,<0.05); the expressions of Iba1 mRNA and CD11b protein expressions were higher than those in the Hegu (LI 4)-Neiguan (PC 6) group (all<0.05); GFAP mRNA and protein expressions were higher apparently than those in the Futu (LI 18) group and the Hegu (LI 4)-Neiguan (PC 6) group (all<0.05).</p><p><b>CONCLUSIONS</b>EA at "Futu" (LI 18) or "Hegu" (LI 4), "Neiguan" (PC 6) relieves the acute neck incisional pain in the rats and its effect may be closely relevant with the down-regulation of the activities of microgliacytes and astrocytes in the spinal cords.</p>

Prospective microglia and brain macrophage distribution pattern in normal rat brain shows age sensitive dispersal and stabilization with development.

The monocytic lineage cells in brain, generally speaking brain macrophage and/or microglia show some dissimilar distribution patterns and disagreement regarding their origin and onset in brain. Here, we investigated its onset and distribution/colonization pattern in normal brain with development. Primarily, early and late embryonic stages, neonate and adult brains were sectioned for routine H/E staining; a modified silver-gold staining was used for discriminating monocytic lineage cells in brain; and TEM to deliver ultramicroscopic details of these cells in brain. Immunofluorescence study with CD11b marker revealed the distribution of active microglia/macrophage like cells. Overall, in early embryonic day 12, the band of densely stained cells are found at the margin of developing ventricles and cells sprout from there dispersed towards the outer edge. However, with development, this band shrunk and the dispersion trend decreased. The deeply stained macrophage like cell population migration from outer cortex to ventricle observed highest in late embryonic days, continued with decreased amount in neonates and settled down in adult. In adult, a few blood borne macrophage like cells were observed through the vascular margins. TEM study depicted less distinguishable features of cells in brain in early embryo, whereas from late embryo to adult different neuroglial populations and microglia/macrophages showed distinctive features and organization in brain. CD11b expression showed some similarity, though not fully, with the distribution pattern depending on the differentiation/activation status of these macrophage lineage cells. This study provides some generalized spatial and temporal pattern of macrophage/microglia distribution in rat brain, and further indicates some intrigue areas that need to be addressed.

Effect of integrin CD11b on liver ischemia/reperfusion injury in mice and underlying mechanism / 中华器官移植杂志

Objective To investigate the role of integrin CD11b in liver ischemia/reperfusion (I/R) injury and its possible mechanism.Method CD11b-/-and WT (C57BL/6) mice were used to establish a 70％ liver warm I/R by clamping the left and median liver lobes for 60 min with vascular micro clamp at 37℃,then the clamp was removed and the abdominal incision was sutured.The blood plasma and liver samples were obtained at different time points (1,3,6,12,24 and 48 h) postreperfusion to assess liver function and cellular injury.Serum ALT and AST levels were determined,and HE staining and TUNEL assay were performed to estimate the severity of liver damage.Tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) were assayed by Reverse transcription polymerase chain reaction (RT-PCR).Kupffer cells were isolated from the live,and the reduced form of nicotinamide-adenine dinucleotide phosphate (NADPH) oxidase activity and active oxygen species (ROS) production were assayed.Result CD11b-/-mice displayed a significantly preserved liver function as represented by lower alanine aminotransferase (ALT) and aspartic transaminase (AST) levels,less histological damage and apoptosis compared to WT mice.Furthermore,TNF-α was decreased and IL-10 mRNA expression was increased in CD11b-/-mice compared to WT mice.Finally,CD11b-/-mice showed decreased activity of NADPH oxidase and less ROS production.Conclusion Integrin CD11 b may regulate the levels of inflammatory (TNF-α) and anti-inflammatory (IL-10) cytokines,enhance the activity of NADPH oxidase in Kupffer cells and enrich the production of ROS,which aggravate liver I/R injury.

Effect of Electroacupuncture at Point Fenglong on the Expressions of Peritoneal Macrophage CD11b and ICAM-1 in Hyperlipidemia Rats / 上海针灸杂志

Objective To investigate the effect of electroacupuncture at point Fenglong(ST40) on the expressions of macrophage surface antigen CD11b and intercellular adhesion molecule-1 (ICAM-1) in hyperlipidemia rats. Method Forty healthy SD rats were randomly allocated to normal control, high fat diet, high fat+common diet, high fat diet and treatment, and high fat+common diet and treatment groups. After 28 days of treatment with electroacupuncture at point Fenglong, blood lipid levels, namely, total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C) and high density lipoprotein cholesterol (HDL-C) contents were measured in every group of rats. The expressions of peritoneal macrophage surface antigen CD11b and ICAM-1 were examined using flow cytometry (FCM) in every group of rats. Result Plasma TC and LDL-C increased significantly in the high fat diet group of rats compared with the normal control group (P0.05). The expression rates of macrophage CD11b and ICAM-1 increased significantly in the high fat diet group of rats compared with the normal group (P<0.01). In the high fat+common diet group of rats, the expression rates of macrophage CD11b and ICAM-1 decreased significantly in comparison with the high fat diet group (P<0.05) but increased significantly in comparison with the normal group (P<0.01). The expression rates of CD11b and ICAM-1 decreased significantly in the high fat diet and treatment group compared with the high fat diet group (P<0.01) and decreased significantly in the high fat+common diet and treatment group compared with the high fat+common diet group (P<0.01) and the high fat diet and treatment group (P<0.01). Correlation analysis showed that CD11b and ICAM-1 levels had a significantly positive correlation (r=0.947, P<0.01). Conclusion Electroacupuncture at point Fenglong can significantly down-regulate blood fat TC and LDL-C levels and the expressions of macrophage CD11b and ICAM-1 in hyperlipidemia rats. It has a certain therapeutic effect on hyperlipidemia.

Influence of eldepryl in proliferation and activation of gliacytes in substantia nigra and striatum in rats with Parkinson’s disease / 吉林大学学报(医学版)

Objective To discuss the influence of eldepryl on the expressions of glial fibrillary acidic protein (GFAP) and cd11b in substantia nigra and striatum in the rats with Parkinson’s disease (PD),and to clarify the regulatory role of eldepryl in the gliacytes.Methods 72 SD rats were randomly divided into control group,PD model group and eldepryl group,and each group was divided randomly into 4 d and 8 d subgroups (n=12)after the success of model preparation.The PD rat models were established by injecting rotenone in subcutaneous.The number of GFAP and cd11b positive cells and the expressions of GFAP and cd11b were detected by immunohistochemistry and Western blotting method.Results The GFAP and cd11b positive cells were all in a resting state in control group, the GFAP-positive cell body was slender and irregular and had elongated protrusions;the cd1 1 b-positive cell body was small and branch-like,and it had more slender protrusions.The GFAP and cd11b positive cells were all in a active state in model group, the GFAP-positive cell body was hypertrophy, the proj ections increased thickening;the cd1 1 b-positive cell body was more bigger, the proj ections were shorter and thicker, and the number was increased.Compared with model group, the GFAP-positive cell body and protrusions were more slender, the CD11b-positive cell body was more smaller,the projections were more slender,and the number was decreased in eldepryl group.There were a small amount of expression of GFAP and cd11b positive cells in substantia nigra and striatum in the rats in control group,and there was no significant difference between 8 d group and 4 d group(P>0.05). The number of GFAP and cd11b positive cells and the protein expression levels were significantly increased in model group compared with control group(P0.05).The number of GFAP and cd11b positive cells and the protein expression levels in eldepryl group were significantly reduced compared with model group(P<0.01);there were less expression in 8 d group compared with 4 d group, and there was significant difference (P<0.05 ). Conclusion There are activation and proliferation of the gliacytes in substantia nigra and striatum in the rats with PD,and eldepryl can inhibit the activation and proliferation of gliacytes.

Effect of resveratrol on proliferation and differentiation in K562 cells / 中国药理学通报

Aim To investigate the effect of resveratrol on proliferation and differentiation in K562 cells. Methods K562 cells were treated with different con-centrations of resveratrol for 6d. The colony number of K562 cells was detected with semi-solid culture assay. Expression of GATA-1 and PU. 1 in K562 cells was re-spectively measured with immunocytochemistry and Western blot. Expression of differentiation related anti-gen, CD11b, CD14 and CD42b, was measured with flowcytometry on K562 cells. Results Resveratrol could significantly decrease the colony number of K562 cells in a dose-dependent manner, and enhance the ex-pression of GATA-1,PU. 1,CD11b, CD14 and CD42b in K562 cells. Conclusion Resveratrol could inhibit the proliferation and induce differentiation of K562 cells via up-regulating the expression of GATA-1 and PU. 1 protein.