Different effects of long-term and short-term repeated restraints on the hematopoietic stem cells in mice / 生理学报

Humans with chronic psychological stress are prone to develop multiple disorders of body function including impairment of immune system. Chronic psychological stress has been reported to have negative effects on body immune system. However, the underlying mechanisms have not been clearly demonstrated. All immune cells are derived from hematopoietic stem cells (HSC) in the bone marrow, including myeloid cells which comprise the innate immunity as a pivotal component. In this study, to explore the effects of chronic psychological stress on HSC and myeloid cells, different repeated restraint sessions were applied, including long-term mild restraint in which mice were individually subjected to a 2 h restraint session twice daily (morning and afternoon/between 9:00 and 17:00) for 4 weeks, and short-term vigorous restraint in which mice were individually subjected to a 16 h restraint session (from 17:00 to 9:00 next day) for 5 days. At the end of restraint, mice were sacrificed and the total cell numbers in the bone marrow and peripheral blood were measured by cell counting. The proportions and absolute numbers of HSC (LinCD117Sca1CD150CD48) and myeloid cells (CD11bLy6C) were detected by fluorescence activated cell sorting (FACS) analysis. Proliferation of HSC was measured by BrdU incorporation assay. The results indicated that the absolute number of HSC was increased upon long-term mild restraint, but was decreased upon short-term vigorous restraint with impaired proliferation. Both long-term mild restraint and short-term vigorous restraint led to the accumulation of CD11bLy6C cells in the bone marrow as well as in the peripheral blood, as indicated by the absolute cell numbers. Taken together, long-term chronic stress led to increased ratio and absolute number of HSC in mice, while short-term stress had opposite effects, which suggests that stress-induced accumulation of CD11bLy6C myeloid cells might not result from increased number of HSC.

Effect of Yangxue Qingnao Granule on the Expression of CD11b in CA1 Region of Hippocampus of Vascular Dementia Rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the effect of Yangxue Qingnao Granule (YQG) on the expression of CD11b in CA1 region of hippocampus of vascular dementia rats, and to explore its regulation on microglias.</p><p><b>METHODS</b>Totally 144 SD rats were randomly divided into the sham-operation group, the vascular dementia model group (model), and the YQG treated group (treated). The vascular dementia rat model was prepared by modified Pulsinelli's four-vessel occlusion. Rats in the sham-operation group and the model group were administered with normal saline -(at the daily dose of 10 mL/kg) by gastrogavage, while those in the treated group were administered with YQG (0.32 g/mL, at the daily dose of 10 mL/kg) by gastrogavage. All administration was performed once per day for 8 successive weeks. The expression of CD11b in CA1 region of hippocampus of vascular dementia rats was detected at week 1, 2, 4, and 8, respectively.</p><p><b>RESULTS</b>Compared with the sham-operation group, the expression of CD11b in CA1 region of hippocampus of vascular dementia rats were significantly enhanced in the model group at each time point (P < 0.01). Compared with the model group, the expression of CD11b in CA1 region of hippocampus of vascular dementia rats significantly decreased in the treated group at each time point (P < 0.01), especially at week 2.</p><p><b>CONCLUSION</b>Obvious activation and proliferation of microglias could be seen in CA1 region of hippocampus of vascular dementia rats, and YQG could inhibit activation and proliferation of microglias.</p>

Effect of Chinese herbal compound Naofucong () on the inflammatory process induced by high glucose in BV-2 cells / 中国结合医学杂志

<p><b>OBJECTIVE</b>To determine the effect of medicated serum of Chinese herbal compound Naofucong (, NFC) on the microglia BV-2 cells viability and the transcription and expression of interleukin-6 (IL-6) and tumor necrosis factor α (TNF-α) in microglia BV-2 cells to further explore the mechanisms underlying the protective effect of NFC on inflammatory process induced by high glucose.</p><p><b>METHODS</b>The microglia BV-2 cells incubated in vitro were divided into different groups: the control group (25 mmol/L glucose), the model group (75 mmol/L glucose), high glucose media containing different dose medicated serum of NFC. After being cultured for 24 h, changes in IL-6 and TNF-α were measured by quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay. The expression of surface marker CD11b of activated microglia was measured by confocal laser scanning microscope and Western blot. Nuclear factor-κB (NF-κB) p-p65 expression was analyzed by Western blot.</p><p><b>RESULTS</b>The model group obviously increased the expression of microglial surface marker CD11b and NF-κB p-p65 (all P<0.01), induced a signifificant up-regulation of release and the mRNA expression of IL-6 and TNF-α (P<0.01 or P<0.05). The medicated serum of NFC could obviously down-regulate the transcription and expression of surface marker CD11 b and NF-κB p-p65 (all P<0.01), and inhibit the mRNA and protein expression (P<0.01 or P<0.05) of inflflammatory cytokines, such as IL-6 and TNF-α, in microglia BV-2 cells cultured with high glucose for 24 h.</p><p><b>CONCLUSIONS</b>The inhibition of microglial activation and IL-6 and TNF-α expression induced by high glucose may at least partly explain NFC therapeutic effects on diabetes-associated cognitive decline diseases. Its underlying mechanism could probably be related to the inhibition of NFC on NF-κB phosphorylation.</p>

Electroacupuncture attenuates spinal nerve ligation-induced microglial activation mediated by p38 mitogen-activated protein kinase / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate whether analgesic effect of electroacupuncture (EA) is affected by p38 mitogen-activated protein kinase (p38 MAPK) on microglia.</p><p><b>METHODS</b>There were two experiments. The experiment 1: 40 male Sprague-Dawley (SD) rats were randomly divided into the normal, surgery, EA and sham EA groups, and the L5 spinal nerve ligation (SNL) on the right side was used to establish neuropathic pain model. EA was applied to bilateral Zusanli (ST36) and Kunlun (BL60) at 24, 48 and 72 h after SNL for 30 min, once per day. The paw withdrawal thresholds (PWTs) were measured before surgery (as base) and at 24, 25, 49 and 73 h after surgery. Phospho-p38 MAPK (p-p38 MAPK), oxycocin-42 (OX-42, marker of microglia), and glial fibrillary acidic protein (GFAP, marker of astrocyte) in bilateral spinal cord dorsal horn (SCDH) were detected by immunofluorescence, respectively. The experiment 2: 40 male SD rats were cannulated for SNL-induced neuropathic pain, and then were randomly divided into the dimethyl sulfoxide (DMSO), EA plus DMSO, 4-(4-fluorophenyl)-2-(4-methylsulfonylpheny)-5-(4-pyridyl)-1H-imidazole (SB203580) and EA plus SB203580 groups. SB203580 (30 nmol/L) was administered 5 min prior to EA treatment. The PWTs and OX-42 in bilateral SCDH were measured as mentioned above.</p><p><b>RESULTS</b>SNL-induced neuropathic pain reduced PWTs and increased the expression of p-p38 MAPK and OX-42 in bilateral lumbar SCDH of rats (P<0.01). Spinal p-p38 MAPK was only co-localized with OX-42 in our study. EA treatment significantly alleviated SNL-mediated mechanical hyperalgesia, and suppressed the expression of p-p38 MAPK and OX-42 in lumbar SCDH (P<0.05 or P<0.01). Intrathecal injection of low dose SB203580 had no influence on PWTs (P>0.05), but significantly inhibited the expression of OX-42 positive cells in bilateral SCDH (P<0.01 or P<0.05). EA plus SB203580 synergistically increased PWTs, and reduced the expression of bilateral spinal OX-42 (P<0.01 or P<0.05).</p><p><b>CONCLUSIONS</b>The central mechanism of EA-induced anti-hyperalgesia may be partially associated with the reduced expression of p-p38 MAPK, and subsequently reducing the activation of OX-42 in neuropathic pain. Therefore, EA may be a new complementary and alternative therapy for neuropathic pain.</p>

Observation of post-MCAO cortical inflammatory edema in rats by 7.0 Tesla MRI / 华中科技大学学报（医学）（英德文版）

This study aimed to investigate inflammatory edema after cerebral ischemia through 7.0T MRI and proton magnetic resonance spectroscopy (MRS). All SD rats were randomly divided into sham operated group and middle cerebral artery occlusion (MCAO)-1 day, -3 day and -7 day groups. MRI scan of the brain was performed on a 7.0 Tesla MRI scanner. The volume of positive signals in the ischemic side was detected by using a T2 weighted spinecho multislice sequence; the changes in the height of water-peak were measured with point resolved spectroscopy (PRESS) sequences; cortical edema was detected by using wet-dry weight method; the degrees of nerve injury were evaluated by Bederson neurological score system; double-labeling immunofluorescence technique was used to explore the molecular mechanisms of post-ischemia cerebral edema. The results showed that high T2WI signals were observed in MCAO-1 day, -3 day and -7 day groups, and the water-peak height and water-peak area of MCAO groups were higher than those of sham operated group (P<0.05). Neurological score results were consistent with the degree of brain edema, and a large number of microglia accumulated in the ischemic cortex. Our results suggested that non-invasive MRI technology with the advantage of high spatial resolution and tissue resolution can comprehensively and dynamically observe inflammatory edema after cerebral ischemia from a three-dimensional space, and contribute to evaluation and treatments in clinic.

Cyclooxygenase-2 blockade inhibits accumulation and function of myeloid-derived suppressor cells and restores T cell response after traumatic stress / 华中科技大学学报（医学）（英德文版）

Myeloid-derived suppressor cells (MDSCs) play a crucial role in T cell dysfunction, which is related to poor outcome in patients with severe trauma. Cyclooxygenase-2 (Cox-2) contributes to immune disorder in trauma and infection via production of prostaglandin E2. However, the role of Cox-2 in the accumulation and function of MDSCs after traumatic stress has not been fully elucidated. In the present study, we treated murine trauma model with NS398, a selective Cox-2 inhibitor. Then the percentages of CD11b+/Gr-1+ cells, proliferation and apoptosis of CD4+ T cells were determined. Arginase activity and arginase-1 (Arg-1) protein expression of splenic CD11b+/Gr-1+ cells, and delayed-type hypersensitivity (DTH) response were analyzed. The results showed that Cox-2 blockade significantly decreased the percentages of CD11b+/Gr-1+ cells in the spleen and bone marrow 48 and 72 h after traumatic stress. NS398 inhibited arginase activity and down-regulated the Arg-1 expression of splenic CD11b+/Gr-1+ cells. Moreover, NS398 could promote proliferation and inhibit apoptosis of CD4+ T cells. It also restored DTH response of traumatic mice. Taken together, our data revealed that Cox-2 might play a pivotal role in the accumulation and function of MDSC after traumatic stress.

The protective role of myeloid-derived suppressor cells in concanavalin A-induced hepatic injury

The mechanism underlying T cell-mediated fulminant hepatitis is not fully understood. In this study, we investigated whether myeloid derived suppressor cells (MDSCs) could prevent the concanavalin A (ConA)-induced hepatitis through suppressing T cell proliferation. We observed an increase in the frequencies of MDSCs in mouse spleen and liver at early stage of ConA treatment, implicating that the MDSCs might be involved in the initial resistance of mice against ConA-mediated inflammation. Subpopulation analysis showed that the MDSCs in liver of ConA-induced mice were mainly granulocytic MDSCs. Adoptive transfer of the bone marrow-derived MDSCs into ConA-treated mice showed that the MDSCs migrated into the liver and spleen where they suppressed T cell proliferation through ROS pathway. In addition, the frequencies of MDSCs in mice were also significantly increased by the treatment with immune suppressor glucocorticoids. Transfer of MDSCs into the regulatory T cell (Treg)-depleted mice showed that the protective effect of MDSCs on ConA-induced hepatitis is Treg-independent. In conclusion, our results demonstrate that MDSCs possess a direct protective role in T cell-mediated hepatitis, and increasing the frequency of MDSCs by either adoptive transfer or glucocorticoid treatment represents a potential cell-based therapeutic strategy for the acute inflammatory disease.

Intrathecal Lamotrigine Attenuates Antinociceptive Morphine Tolerance and Suppresses Spinal Glial Cell Activation in Morphine-Tolerant Rats

Glial cells play a critical role in morphine tolerance, resulting from repeated administration of morphine. Both the development and the expression of tolerance are suppressed by the analgesic lamotrigine. This study investigated the relationship between the ability of lamotrigine to maintain the antinociceptive effect of morphine during tolerance development and glial cell activation in the spinal cord. In a rat model, morphine (15 microg) was intrathecally injected once daily for 7 days to induce morphine tolerance. Lamotrigine (200 microg) was co-administered with morphine either for 7 days or the first or last 3 days of this 7 day period. Thermal nociception was measured. OX-42 and GFAP immunoreactivity, indicating spinal microglial and astrocytic activation were evaluated on day 8. Tolerance developed after 7 days of intrathecal morphine administration; however, this was completely blocked and reversed by co-administration of lamotrigine. When lamotrigine was coinjected with morphine on days 5-7, the morphine effect was partially restored. Glial cell activation increased with the development of morphine tolerance but was clearly inhibited in the presence of lamotrigine. These results suggest that, in association with the suppression of spinal glial cell activity, intrathecally coadministered lamotrigine attenuates antinociceptive tolerance to morphine.

Microglial P2X7 receptor expression is accompanied by neuronal damage in the cerebral cortex of the APPswe/PS1dE9 mouse model of Alzheimer's disease

The possibility that P2X7 receptor (P2X7R) expression in microglia would mediate neuronal damage via reactive oxygen species (ROS) production was examined in the APPswe/PS1dE9 mouse model of Alzheimer's disease (AD). P2X7R was predominantly expressed in CD11b-immunopositive microglia from 3 months of age before Abeta plaque formation. In addition, gp91phox, a catalytic subunit of NADPH oxidase, and ethidium fluorescence were detected in P2X7R-positive microglial cells of animals at 6 months of age, indicating that P2X7R-positive microglia could produce ROS. Postsynaptic density 95-positive dendrites showed significant damage in regions positive for P2X7R in the cerebral cortex of 6 month-old mice. Taken together, up-regulation of P2X7R activation and ROS production in microglia are parallel with Abeta increase and correlate with synaptotoxicity in AD.

Role of IL-1alpha in Cisplatin-Induced Acute Renal Failure in Mice

BACKGROUND/AIMS: For unknown reasons, caspase-1 -/- mice, protected against cisplatin-induced acute renal failure (ARF), are deficient in interleukin (IL)-1alpha. We thus asked whether IL-1alpha deficiency underlies the mechanism of protection against cisplatin-induced ARF in these mice. METHODS: Cisplatin (30 mg/kg) was injected intraperitoneally into wild-type C57BL/6 mice to produce a cisplatin-induced model of ARF. IL-1alpha was measured in control vehicle- and cisplatin-treated wild-type animals. We also examined whether IL-1alpha -/- mice were similarly protected against cisplatin-induced ARF. Additionally, infiltration of CD11b- and CD49b-positive cells, as markers of macrophages, natural killer, and natural killer T cells (pan-NK cells), was investigated in wild-type and IL-1alpha -/- mice. RESULTS: Compared with vehicle-treated mice, renal IL-1alpha increased in cisplatin-treated wild-type mice beginning on day 1. IL-1alpha -/- mice were shown to be protected against cisplatin-induced ARF. No significant difference in the infiltration of neutrophils or CD11b- and CD49b-positive cells were observed between wild-type and IL-1alpha -/- mice. CONCLUSIONS: Mice deficient in IL-1alpha are protected against cisplatin-induced ARF. The lack of IL-1alpha may explain, at least in part, the protection against cisplatin-induced ARF observed in caspase-1 -/- mice. Investigation of the protective mechanism (s) in IL-1alpha -/- mice in cisplatin-induced ARF merits further study.

Effect of C/EBPalpha on the monocytic differentiation of HL60 cells induced by NSC67657 / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To figure out the function of C/EBPalpha in the monocytic differentiation of HL60 cells induced by a new steroidal drug NSC67657.</p><p><b>METHODS</b>The differentiation of HL60 cells was induced by NSC67657, and the cell surface antigen CD14 expression was detected by flow cytometry. The gene and protein expressions of CCAAT enhancer binding protein alpha (C/EBPalpha) before and after the induction of cell differentiation were determined by RT-PCR and Western blot. Eukaryotic expressing vector pDsRed-ICAT was constructed and transfected into HL60 cells, and its expression was verified. The effect of C/EBPalpha overexpression in HL60 cells was assessed by MTT assay, Wright's staining and flow cytometry before and after NSC67657 transfection.</p><p><b>RESULTS</b>HL60 cells could be induced into monocytes by 10 micromol/L ATRA within 5 days, and the coverage of CD14 positive cells reached 93.9% after 5 days of drug treatment. The eukaryotic expressing vector was successfully constructed, and over 90% positive clones were obtained after screening by G418 and electrotransfection. The results of proliferative analysis, chemical staining, ultrastructural observation, and CD11b detection confirmed that HL60 cells could be induced into granulocytic differentiation by overexpression of C/EBPalpha protein. Moreover, in the drug treatment group, transfected cells could not be induced into monocytic differentiation, and their granulocytic differentiation was also inhibited.</p><p><b>CONCLUSION</b>The monocytic differentiation of HL60 cells induced by NSC67657 may not be via the regulation by C/EBPalpha protein-mediated signal transduction. However, the overexpression of CEBPalpha may inhibit the process of NSC67657-induced monocytic differentiation in HL60 cells.</p>

Blood hibernation: a novel strategy to inhibit systemic inflammation and coagulation induced by cardiopulmonary bypass / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Inflammation and coagulation are two intimately cross-linked defense mechanisms of most, if not all organisms to injuries. During cardiopulmonary bypass (CPB), these two processes are activated and interact with each other through several common pathways, which may result in subsequent organ dysfunction. In the present study, we hypothesized that the addition of nitric oxide, prostaglandin E1 (PGE1), and aprotinin to the systemic circulation, hereby referred to as blood hibernation, would attenuate the inflammation and coagulation induced by CPB.</p><p><b>METHODS</b>Thirty adult mongrel dogs were equally divided into five groups, anesthetized and placed on hypothermic CPB (32 degrees C). Each group received respectively the following treatments: (1) inhalation of 40 ppm nitric oxide; (2) intravenous infusion of 20 ng x kg(-1) x min(-1) of PGE1; (3) 80,000 kallikrein inhibitor units (KIU)/kg of aprotinin; (4) the combination of all three agents (blood hibernation group); and (5) no treatment (control group) during CPB. Activation of leukocyte, platelet, endothelial cell, and formation of thrombin were assessed after CPB.</p><p><b>RESULTS</b>As compared with the other four groups, leukocyte counts were higher, while plasma elastase, interleukin-8, CD11b mRNA expression, myeloperoxidase activities and lung tissue leukocyte counts were lower in the blood hibernation group (P < 0.05 versus other four groups after CPB). Plasma prothrombin fragment (PTF)1+2, and platelet activation factors were lower, while platelet counts were higher in the blood hibernation group (P < 0.05 versus other four groups at 6 and 12 hours after CPB). Electron microscopy showed endothelial pseudopods protrusion, with cell adherence in all four groups except the blood hibernation group where endothelial cells remained intact.</p><p><b>CONCLUSION</b>Blood hibernation, effected by the addition of nitric oxide, PGE1 and aprotinin to the circulating blood during extra-corporeal circulation, was observed to attenuate the inflammation and coagulation induced by cardiopulmonary bypass, most likely by inhibiting the important common intermediates between the two cross-linked processes.</p>

The effect of ATRA-induced leukemic cell differentiation on Brd7 gene expression in leukemia cell lines / 中国实验血液学杂志

This study was purposed to investigate the relationship between brd7 gene and differentiation of leukemia cells and the role of brd7 gene in differentiation of leukemia cells. The HL-60 and K562 cell lines were induced by all-trans retinoic acid (ATRA) for 7 days, then the cell morphologic change was observed under inverted microscope with Wright-Giema staining, the expression level of CD11b was detected by flow cytometry for evaluating cell differentiation level, the expression changes of BRD7 protein before inducing differentiation and in process of cell differentiation were determined by Western blot. The results showed that ATRA could inhibit the proliferation and induce differentiation of HL-60 cells, but no differentiation in K562 cells was induced by ATRA. The level of CD11b expression in HL-60 cells was up-regulated gradually during ATRA-induced cell differentiation. The expression of BRD7 protein increased markedly along with differentiation of HL-60 cells towards granulocytes. However, BRD7 protein did not significantly alter in K562 cells in which inducing differentiation was not found. It is concluded that brd7 gene expression enhances as the HL-60 cells differentiate, underlying which the mechanism remains to clarify.

Effect of Kidney-Tonifying and Blood-Promoting Recipe on the expression of CD11b/CD18 and Bcl-2/Bax in aged patients with kidney deficiency and blood stasis syndrome / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the effect of Kidney-Tonifying plus Blood-Promoting Recipe on the expression of CD11b/CD18 and Bcl-2/Bax in elderly patients with kidney deficiency and blood stasis syndrome.</p><p><b>METHODS</b>Sixty elderly patients with kidney deficiency and blood stasis syndrome were randomized into two groups. Patients in the treatment group received Kidney-Tonifying plus Blood-Promoting Recipe, and those in the control group receive no treatment. The expression of CD11b/CD18, Bcl-2/Bax, D-Dimeride, CD62p, PAC-I and the rate of platelet aggregation in peripheral blood leukocytes before and after the treatment were examined using flow cytometry in both groups.</p><p><b>RESULTS</b>The Recipe significantly decreased the levels of CD11b/CD18, D-Dimeride, CD62p, PAC-I and the rate of platelet aggregation (P<0.01), and increased the levels of Bcl-2/Bax (P<0.01).</p><p><b>CONCLUSION</b>Kidney-Tonifying plus Blood-Promoting Recipe regulates CD11b/CD18 and Bcl-2/Bax expression in blood leukocytes and improves microcirculatory status, which can be one of the mechanisms underlying its therapeutic effect in elderly patients.</p>

Alteration of the expression ratio between WT1 gene and its isomers during all-trans retinoic acid-induced differentiation of HL-60 cells / 中华病理学杂志

<p><b>OBJECTIVE</b>To explore expression ratio alteration between WT1 gene and its isomers during differentiation of leukemia cell line HL-60 induced by all-trans retinoic acid (ATRA) and the relationship existed between them.</p><p><b>METHODS</b>The degree of cellular maturation was verified by NBT reduction test and immunophenotyping. Expression of unspliced WT1, WT1 (17AA+) and WT1 (KTS+) were determined by real-time fluorescence quantitative RT-PCR during the induced differentiation of HL-60 cells. The relative ratio of four splicing variants WT1 (+/+), WT1 (+/-), WT1 (-/+), WT1 (-/-) were analyzed.</p><p><b>RESULTS</b>During the induced differentiation of HL-60 cells, NBT reduction rate and CD11b positive rate increased significantly (P < 0.05 and P < 0.001, respectively). The expression of WT1 gene decreased from (4.17 +/- 2.21) x 10(-3) at 0 hour to (7.53 +/- 2.30) x 10(-4) at the 96th hour. The ratio of 17AA+decreased from 0.60 +/- 0.05 at 0 hour to 0.42 +/- 0.08 at the 96th hour. The ratio of KTS+ decreased from 0.53 +/- 0.08 at 0 hour to 0.41 +/- 0.04 at the 96th hour. The ratio of WT1 (+/+) decreased gradually from 0.32 +/- 0.06 at 0 hour to 0.17 +/- 0.03 at the 96th hour. Change of ratios of other two isomers not significant.</p><p><b>CONCLUSIONS</b>During the induced differentiation of HL-60 cells, WT1 gene expression decreases gradually. The phenotype of the majority of uninduced HL-60 cells is WT1 (+/+), in contrast to WT1 (-/-) phenotype after the induction of cell differentiation, indicating that WT1 gene may participate in the regulation of hematopoietic cell differentiation through modulation of the expression ratios of its four spliced variants.</p>

Evaluation of neutrophil CD 64 and CD 11b expression as early predictors of sepsis severity and outcome

CD64, the high affinity Fcy receptor 1, and CDllb, an alpha-subunit of the beta[2] integrin adhesion molecule, are specific neutrophil-surface antigens activated in response to systemic inflammation, therefore, they could potentially be used as early predictors of sepsis and the associated mortality in patients with systemic inflammatory response syndrome [SIRS]. Sixty-one SIRS patients with clinically suspected infection were enrolled and subjected to sepsis work-up, within the first 24 hours of sepsis onset, including complete blood count, blood culture, serum C-reactive protein [CRP] in addition to flow cytometric analysis of CD64 and GD11b. Patients were classified prospectively on the basis of clinical observation and blood culture results into two groups: SIRS with sepsis group [n=36] and SIRS without sepsis group [n=25] served as patient control. According to outcome, patients with sepsis were classified after a follow-up period up to 28 days after inclusion into two groups: survivors [n=29] and non-survivors [n=7]. A highly significant increase of neutrophil CD64 and CD11b expression was detected in the sepsis group as compared to the non-infected group. CD64 and CDllb expression had the best diagnostic performance for prediction of early-onset sepsis. Expression of CD64 at a cut-off value of 49% had 88.9% sensitivity, 92% specificity and 90.2% efficacy, while CD1 Ib expression at a cut-off value of 71% had 86.1% sensitivity, 84% specificity and 85.2% efficacy. Combined use of both markers yielded 91.7% sensitivity, 96% specificity and 93.4% efficacy. Sepsis survivors showed significantly lower expression of CD64 and CDllb as compared to non-survivors. An optimal cut-off value of 70% expression for CD64 predicted mortality with 100% sensitivity, 96.6% specificity and 97.2% efficacy. Meanwhile, a cut-off value of 86% for CDllb predicted mortality with 85.7% sensitivity, 93.1% specificity and 91.7% efficacy. Assessment of neutrophil CD64 and CDllb expression is superior to standard laboratory tests for early detection of sepsis, within the first 24 hours of sepsis onset, before the evolution of clinical signs which would facilitate therapeutic decisions. Prediction of outcome is an additional advantage borne by these two biomarkers

CD11b expression in neutrophils and lymphocytes of children with systemic inflammatory response syndrome / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the significance of CD11b expression in neutrophils and lymphocytes in children with systemic inflammatory response syndrome (SIRS).</p><p><b>METHODS</b>CD11b expression in neutrophils and lymphocytes was measured using flow cytometry in 36 children with SIRS (SIRS group) and 28 children with infectious disease but without SIRS (control group). The sensitivity and specificity of neutrophil CD11b for diagnosis of SIRS were evaluated.</p><p><b>RESULTS</b>During the acute phase, an increased CD11b expression in neutrophils (96.7+/-8.1%) was observed in the SIRS group compared with the control group (85.1+/-5.1%) (p<0.05). Using neutrophil CD11b expression >92.2% as a cut-off value for diagnosis of SIRS, the sensitivity and the specificity were 97.2 % and 92.9% respectively. Lymphocytic CD11b expression in the SIRS group (13.4+/-8.6%) was lower than that in the control group (19.2+/-6.4%) in the acute phase (p<0.05). In the SIRS group, lymphocytic CD11b expression was remarkably suppressed in the severe sepsis subgroup (7.27+/-3.04%), showing significantly decreased expression compared with the non-infectious subgroup (19.3+/-2.9%) and the sepsis subgroup (15.9+/-12.5%) (p<0.01). In the convalescence stage lymphocytic CD11b expression in the SIRS group was similar to that in the control group.</p><p><b>CONCLUSIONS</b>CD11b expression in neutrophils may serve as a reliable indicator for diagnosis of SIRS. The down-regulation of lymphocytic CD11b expression might be a signal of the condition aggravation in children with SIRS.</p>

CD56 and CD11b antigen expressions in patients with acute monocytic leukemia and the clinical implications / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expressions of cell surface differentiation antigen CD56 and CD11b antigen in acute monocytic leukemic (AML-M(5)) cells and their clinical significance.</p><p><b>METHODS</b>A total of 113 cases of de nove adult AML-M(5) were examined genetically and immunologically using G-banding technique, interphase fluorescence in situ hybridization (I-FISH) and flow cytometry immunophenotyping, and the results were analyzed in relation to their clinical data.</p><p><b>RESULTS</b>Of the 113 cases, the expression rates of CD56 and CD11b was 28.32% and 73.45%, respectively. The CD56(+) patients had high CD11b expression, and the expression levels of CD11b and CD56 were positively correlated (P<0.05). The incidence of karyotypic abnormalities was 48.57% (55 cases) in these patients, including 25 (22.12%) with 11q23 aberrations. Twenty-five cases were positive for MLL gene abnormalities as found by I-FISH analysis. Compared with the patients positive for both CD56 and CD11b, those negative for both CD56 and CD11b showed increased peripheral blood white blood cell (WBC) count and also increased blast and progenitor cells in the bone marrow (P<0.05); the former patients often had karyotypic abnormalities, commonly involving 11q23 aberrations (P<0.05), whereas the latter patients presented more likely with extramedullary infiltration and refractory leukemia (P<0.01) with lowered complete remission rate and shortened median survival time (P<0.01). CD56-positive patients were more likely to have karyotypic abnormalities and refractory leukemia than CD11b-postive patients (P<0.05), but the peripheral blood WBC counts, bone marrow blast and progenitor cells, extramedullary infiltration, complete remission rate or median survival time showed no significant differences between them (P>0.05).</p><p><b>CONCLUSION</b>AML-M(5) patients with CD56 positivity and high expression of CD11b often have aberrant karyotypes, commonly involving 11q23/MLL gene abnormality. These patients frequently develop extramedullary infiltration and refractory leukemia often with poor prognosis.</p>

Effects of icariin on inflammation model stimulated by lipopolysaccharide in vitro and in vivo / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To evaluate the anti-inflammatory effects of icariin, from aspects of pro-inflammatory cytokines, inflammatory mediators and adhesion molecules.</p><p><b>METHODS</b>Mouse inflammation model in vitro was established by stimulating macrophage cell line RAW264. 7 with lipopolysaccharide (LPS); and the inflammation model in vivo was established by stimulating C57BL/6J mouse with LPS. Taking dexamethasone as the positive control, both models were treated with icariin, and the cell viability in model mice was detected with CCK-8 kit; tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6) in cell culture medium and serum were detected by ELISA; nitric oxide (NO) in cell culture medium by Griess Reagent method; CD11b expression on the surface of neutrophil in mice by flow cytometry, and pulmonary inflammatory cell infiltration in mice by pathological section as well.</p><p><b>RESULTS</b>in vitro studies showed that icariin at the doses of 1 microg/mL, 10 microg/mL and 100 microg/mL, all displayed no cytotoxicity (P < 0.01); 10 microg/mL and 100 microg/mL icariin effectively lowered the levels of TNF-alpha and IL-6 (P < 0.01) in medium; and 100 microg/mL icariin significantly reduced level of NO (P < 0.01) in medium. in vivo studies showed that icariin at the dose of 20 mg/kg significantly lowered serum TNF-alpha and IL-6 levels (P < 0.01), reduced the average fluorescence intensity of adhesion molecules CD11b (P < 0.01), and alleviated pulmonary inflammatory cell infiltration.</p><p><b>CONCLUSION</b>Icariin is a safe and effective natural anti-inflammatory drug, its partial mechanism is possible the multiple links intervention on pro-inflammatory cytokines (TNF-alpha, IL-6), inflammatory mediators (NO) and adhesion molecules (CD11b).</p>

Triptolide protects against 1-methyl-4-phenyl pyridinium-induced dopaminergic neurotoxicity in rats: implication for immunosuppressive therapy in Parkinson's disease / 神经科学通报·英文版

<p><b>OBJECTIVE</b>Neuroinflammation with microglial activation has been implicated to have a strong association with the progressive dopaminergic neuronal loss in Parkinson's disease (PD). The present study was undertaken to evaluate the activation profile of microglia in 1-methyl-4-phenyl pyridinium (MPP+)-induced hemiparkinsonian rats. Triptolide, a potent immunosuppressant and microglia inhibitor, was then examined for its efficacy in protecting dopaminergic neurons from injury and ameliorating behavioral disabilities induced by MPP+.</p><p><b>METHODS</b>The rat model of PD was established by intranigral microinjection of MPP+. At baseline and on day 1, 3, 7, 14, 21 following MPP+ injection, the degree of microglial activation was examined by detecting the immunodensity of OX-42 (microglia marker) in the substantia nigra (SN). The number of viable dopaminergic neurons was determined by measuring tyrosine hydroxylase (TH) positive neurons in the SN. Behavioral performances were evaluated by counting the number of rotations induced by apomorphine, calculating scores of forelimb akinesia and vibrissae-elicited forelimb placing asymmetry.</p><p><b>RESULTS</b>Intranigral injection of MPP+ resulted in robust activation of microglia, progressive depletion of dopaminergic neurons, and ongoing aggravation of behavioral disabilities in rats. Triptolide significantly inhibited microglial activation, partially prevented dopaminergic cells from death and improved behavioral performances.</p><p><b>CONCLUSION</b>These data demonstrated for the first time a neuroprotective effect of triptolide on dopaminergic neurons in MPP+-induced hemiparkinsonian rats. The protective effect of triptolide may, at least partially, be related to the inhibition of MPP+-induced microglial activation. Our results lend strong support to the use of immunosuppressive agents in the management of PD.</p>