Novel mutations of ITGB2 induced leukocyte adhesion defect type 1 / 中华儿科杂志

Objective@#To investigate the pathogenic mechanism of two novel ITGB2 mutations in leukocyte adhesion defect type 1 (LAD1).@*Methods@#The clinical history and blood sample of an 11 years old patient admitted to Affiliated Hospital of Qingdao University in August 2014 were collected. Expression of CD18 (encoded by ITGB2) was analyzed by flow cytometry. Novel ITGB2 mutations were identified by next-generation sequencing technology and confirmed by Sanger sequencing. The functional effect of ITGB2 mutations was detected by PolyPhen2. Expression vectors of both wild type and mutant ITGB2 were constructed and transfected into mammalian cells for analysis of protein stability and subcellular location.@*Results@#The symptoms of the patient (recurrent infections, lowered alveolar ridge and hypodontia) supported the diagnosis of LAD1. Expression of CD18 on the leukocytes was significantly decreased (0.2%) compared with the control samples from the parents (paternal: 99.0%; maternal: 99.1%). The patient was identified to be compound heterozygous for ITBG2 c.954del G (novel mutation) and c.1802C>A (paternal originated). ITGB2 c.954 del G was confirmed to be a harmful frameshift mutation; ITGB2 c.1802C>A was also predicted to be harmful. In terms of protein stability. There was no significant difference between mutant D18 and wild type. However, subcellular location analysis showed the mutant D18 could not locate on cell membrane.@*Conclusion@#The compound heterozygous of ITGB2 mutations (c.954del G and c.1802C>A) decreases the expression and impairs the location of CD18 on leukocytes, which leads to LAD1.

The effect of hypertonic sodium chloride in the treatment of patients with traumatic hemorrhagic shock and the effects of NO and neutrophil surface CD 18 in serum / 国际检验医学杂志

Objective To study the effect of hypertonic sodium chloride in the treatment of patients with traumatic hemorrhagic shock and the effects of NO and neutrophil surface CD 18 on the serum.Methods 120 patients with traumatic hemorrhagic shock were admitted to the hospital from December 2013 to December 2016,and randomly divided into the observation group and the control group.The control group was given nal-oxone injection,while the observation group was given hypertonic Sodium Chloride Solution.The two groups were compared before and after treatment,hemoglobin concentration,total infusion,24 h mortality,recovery time and adverse reactions,and the patients′hemodynamic and NO and CD18 levels were measured on the ser-um.Results After 24 h treatment,the observation group total infusion was(1 203.13 ± 117.82)mL,the re-covery time was(60.73 ± 5.63)min,24 h mortality rate of 5.00% was significantly lower than the control group total infusion(1 672.38 ± 123.64)mL,the recovery time(71.82 ± 6.19)min,24 h death rate 16.67%, hemoglobin concentration(91.24 ± 5.71)g/L higher than that of the control group(79.45 ± 6.18)g/L,the difference was statistically significant(P<0.05).Before treatment,there was no difference in the contents of NO and CD18 between the two groups.After treatment,two groups of patients with serum NO,CD18 content significantly decreased,but the observation group NO(20.27 ± 6.65)mol/L,CD18(41.67 ± 13.24)ng/mL were significantly lower than the control group NO(29.12 ± 8.23)mol/L,CD18(52.64 ± 13.51)ng/mL,the difference were statistically significant(P<0.05).Before treatment,two groups of patients with hemodynam-ic indexes of arterial and venous pressure,no difference in heart rate(P> 0.05);after treatment,the two groups of patients with arterial and venous pressure and heart rate were improved,but the change index of the observation group than in the control group was stable,the difference was statistically significant(P<0.05). The complication rate of the observation group was 10%(6/60),which was significantly lower than that of the control group(15/60),and the difference was statistically significant(P<0.05).Conclusion The infiltra-tion of sodium chloride intravenous infusion could significantly decreased CD 18 and NO levels in the serum of the patients,the patients with stable hemodynamics,reduced the incidence of complications,worthy of clinical reference.

Immunomodulatory effects of selected Malaysian plants on the CD18/11a expression and phagocytosis activities of leukocytes

Objective: To investigate the effects of 20 methanolic extracts from Malaysian selected plants on CD18/11a expression and phagocytosis activity of leukocytes using flow cytometry analysis. Methods: The effects of methanolic extracts on CD18/11a expression and phagocytosis of leukocytes were measured by labelling the cells with CD18-fluorescein isothiocyanate and ingestion labelled with Escherichia coli-fluorescein isothiocyanate and then analyzed using flow cytometer. Results: About 12 out of 20 methanolic extracts of selected Malaysian medicinal plants significantly (P≤0.05) inhibited the CD18/11a expression of leukocytes at both concentrations of 6.25 μg/mL and 100 μg/mL in dose dependent manner. The most active inhibitory was shown in Citrus aurantifolia (Christm.) Swingle and Alpinia galangal (L.) Willd. at dosage 100 μg/mL. Moreover, the Orthosiphon aristatus (Blume) Miq (O. aristatus). showed the highest stimulatory activity at the concentration of 100 μg/mL. Other than that, four plant extracts significantly (P≤0.05) rose the phagocytosis activities of leukocytes in dose dependent manner. However, Annona muricata L. and O. aristatus showed the highest stimulated activities at the 100 μg/mL concentration. Conclusions: The results suggest that methanolic extracts of Citrus aurantifolia, Alpinia galangal, O. aristatus and Annona muricata are able to modulate innate immune system and can potentially be recognized as therapeutic agents for modulating immune system.

Effect of Rho kinase combined with Sheng Mai injection on TNF-α, IL-10, ICAM-1 and CD11/CD18 in patients with Ⅱ°superficial burn / 中国生化药物杂志

Objective To investigate effect of Rho kinase combined with Sheng Mai injection on tumor necrosis factor alpha ( TNF-α) , interleukin-10 (IL-10), intercellular adhesion molecule-1 (ICAM-1) and adhesion molecule CD11/CD18 in patients withⅡ°superficial burn.Methods 48 cases patients withⅡ°superficial burn were collected and divided into experimental group and control group, according to different therapy, 24 cases in each group.The two groups were given symptomatic treatment such as anti-infectious, keep warm, anti-shock, pain-relieve and wound-healing treatment, the control group were given Rho kinase on the basis of symptomatic treatment, and experimental group were given Sheng Mai injection on the basis of control group, for a course of two weeks.The serum TNF-α, IL-10, ICAM-1 and CD11/CD18 levels were compared after treatment. ResuIts After treatment, the serum TNF-α, IL-10, ICAM-1 and CD11/CD18 levels after treatment were lower than those before treatment in two groups (P<0.05).Compared with control group, the levels of TNF-α, IL-10, ICAM-1 and CD11/CD18 were lower than those of experimental group (P<0.05).ConcIusion Rho kinase combined with Sheng Mai injection could significantly reduce the levels of TNF-α, IL-10, ICAM-1 and CD11/CD18 in patients with Ⅱ°superficial burn.

Role of NT-κB in monocytes-induced HK-2 cells transdifferentiation / 中华肾脏病杂志

Objective To investigate the effects of monocytes on phenotypic changes of human proximal tubular HK-2 cells and the mechanism.Methods Monocytes were co-cultured with HK-2 cells.Morphological changes of HK-2 cells were detected by inverted phase contrast microscope.Expressions of E-cadherin,α-SMA and fibronectin were assessed by RT-PCR,Western blotting and immunocytochemical staining.Flow cytometry techniques was applied to evaluate intercellular cell adhesion molecule-1 (ICAM-1) expression on HK-2 cells.The intracellular signal was investigated by gene microarr ay.Results The typical epithelial cell morphology of HK-2 cells disappeared after co-culture with monocytes,accompanied by decreased E-cadherin expression and increased α-SMA and fibronectin expression (all P＜0.05).The expression of ICAM-1 on HK-2 cells was increased by monocytes stimulation.Interestingly,administration of CD18 antibody directly inhibited the phenotypic change of HK-2 cells.Furthermore,NF-κB signaling might be critical in mediating this process,and blockade of this signaling pathway could inhibit 1CAM-1 expression and epithelial mesenchymal transition (EMT) formation.Conclusion Monocytes can directly induce EMT of HK-2 cells via up-regulating ICAM-1 through NF-κB signaling pathway.

Expression of beta 2 integrin (CD18) in embryonic mouse and chicken heart

Integrins are heterodimeric receptors composed of á and â transmembrane subunits that mediate attachment of cells to the extracellular matrix and counter-ligands such as ICAM-1 on adjacent cells. â2 integrin (CD18) associates with four different á (CD11) subunits to form an integrin subfamily, which has been reported to be expressed exclusively on leukocytes. However, recent studies indicate that â2 integrin is also expressed by other types of cells. Since the gene for â2 integrin is located in the region of human chromosome 21 associated with congenital heart defects, we postulated that it may be expressed in the developing heart. Here, we show the results from several different techniques used to test this hypothesis. PCR analyses indicated that â2 integrin and the áL, áM, and áX subunits are expressed during heart development. Immunohistochemical studies in both embryonic mouse and chicken hearts, using antibodies directed against the N- or C-terminal of â2 integrin or against its á subunit partners, showed that â2 integrin, as well as the áL, áM, and áX subunits, are expressed by the endothelial and mesenchymal cells of the atrioventricular canal and in the epicardium and myocardium during cardiogenesis. In situ hybridization studies further confirmed the presence of â2 integrin in these various locations in the embryonic heart. These results indicate that the â2 integrin subfamily may have other activities in addition to leukocyte adhesion, such as modulating the migration and differentiation of cells during the morphogenesis of the cardiac valves and myocardial walls of the heart.

Resistance of melanized yeast cells of Paracoccidioides brasiliensis to antimicrobial oxidants and inhibition of phagocytosis using carbohydrates and monoclonal antibody to CD18

Paracoccidioides brasiliensis, a thermal dimorphic fungal pathogen, produces a melanin-like pigment in vitro and in vivo. We investigated the involvement of carbohydrates and monoclonal antibody to CD18, on phagocytosis inhibition, involving macrophage receptors and the resistance of melanized fungal cells to chemically generated nitric oxide (NO), reactive oxygen species (ROS), hypochlorite and H2O2. Our results demonstrate that melanized yeast cells were more resistant than nonmelanized yeast cells to chemically generated NO, ROS, hypochlorite and H2O2, in vitro. Phagocytosis of melanized yeast cells was virtually abolished when mannan, N-acetyl glucosamine and anti-CD18 antibody were added together in this system. Intratracheal infection of BALB/c mice, with melanized yeast cells, resulted in higher lung colony forming units, when compared to nonmelanized yeast cells. Therefore, melanin is a virulence factor of P. brasiliensis.

Influence of hypoxia-inducible factor-1α on the expression of neutrophil CD18 and leukocyte adhesion in diabetic rats / 中华眼底病杂志

Objective To observe the influence of the expression of CD18 on the neutrophile and the leukocyte adhesion to retinal vascular endothelium by hypoxia-inducible factor-1 alpha (HIF-1α) in early diabetic retinopathy rats. Methods Male Sprague-Dawley rats received intraperitoneal injection of streptozotocin to induce diabetes model. 18 diabetic rats were divided into 3 groups randomly after 2 months of diabetes induction, including diabetic group (group B), HIF-1α anti-sense oligonucleotides (ASODN) injection group (group C) and HIF-1α sense oligonueleotides (SODN) injection group (group D), the age and weigh matched health rats were chosen as control group (group A), with 6 rats in each group. Then group A and B rats received 5 % glucose solution caudalis veins injection, group C and group D rats received HIF-1α ASODN and HIF-1α SODN caudalis veins injection, respectively(0.25 mg/kg). The level of CD18 on the neutrophil isolated from the peripheral blood was measured by flow cytometry. Retinal leukostasis was quantified with acridine orange leukocyte fluorography. Results The percentage of CD 18 positive neutrophil cell was (44.93±3. 60)% in group B, (18.66±1.52)% in group A, (31.66±4.72)% in group C,(51.00±5.66)% in group D. Compared with each other groups,the differences are statistically significant (F= 42. 46, P<0.001). The number of positive staining cells of retinal leukocyte was (46.16±10.68)in group A, (133.83±20.43)in group B, (99.83±9.28)in group C, (121.33±10. 23) in group C. Compared group B with group C,the number of positive staining cells raised about 2.89 times;compared group B with group C and D,the differences are statistically significant (P= 0.12, 95% confidence interval -3.69～28. 69 ). Conclusions In vivo, HIF-1α can decreased the expression of CD18 on neutrophils from diabetic rats' peripheral blood and the collection of retinal leukos- tasis in the diabetic animals. HIF-1α may serve as a therapeutic target for the treatment and/or prevention of early diabetic retinopathy.

Methotrexate inhibits integrin adhesion molecules in the mouse model of pleurisy induced by carrageenan

The aim of this work was to analyze the effect of methotrexate (MTX) upon leukocyte migration and expression of adhesion molecules CD11a/CD18 in the lung, 4 and 48 h after inflammation induction by carrageenan in mice. The results showed that MTX significantly decreased leukocyte influx and CD11a expression in the lung at 4 and 48 h of pleurisy (P < 0.01). MTX also inhibited CD18 expression at 4 h but not 48 h of pleurisy (P < 0.01). These results proved that MTX at the studied doses had important anti-inflammatory properties, acting primarily on leukocyte migration from the pleural cavity to the lung via inhibition of CD11a/CD18 expression in the mouse model of inflammation.

O modelo experimental da pleurisia induzida pela carragenina, em camundongos é caracterizado pelo aumento da migração de leucócitos às custas de neutrófilos 4 h após a indução da inflamação pela carragenina na cavidade pleural de camundongos.. Após 48 h da indução da inflamação ocorre aumento de leucócitos do tipo mononucleares. O objetivo deste trabalho foi avaliar o efeito do metotrexato (MTX) sobre a migração de leucócitos, e a expressão de moléculas de adesão (CD11a/CD18), no pulmão 4 e 48 h após a inflamação induzida pela carragenina.Os resultados demonstraram que o MTX inibiu significativamente o influxo de leucócitos e a expressão de CD11a no pulmão 4 h e 48 h após a inflamação induzida pela carragenina (P < 0.01). O MTX inibiu a expressão de CD18 no pulmão 48 h após, mas não 4 h após esta resposta inflamatória (P < 0.01). Estes resultados demonstram que o MTX, nas doses estudadas, possui importante efeito antiinflamatório agindo principalmente sobre o influxo de leucócitos da cavidade pleural para os pulmões, via inibição da expressão de moléculas de adesão do tipo CD11a/CD18, no modelo da pleurisia induzida pela carragenina, em camundongos.

The expressions of ICAM-1 and VCAM-1 in HIE neonatal rabbits / 重庆医科大学学报

Objective:Searching after the effects of the expressions the neonatal rabbits with chronic intrauterine anoxemia give to neutrophil and conglutinated molecule.Method:Choosing 15 Zelanian rabbits that have been in afternoon pregnancy,we divide them into three stochastic contratest groups by chance:the control group,the anoxic group and the group interfered with Chinese traditional medicine.Then,we contrast the expressions and changes of these groups with CD11b/CD18,ICAM-1 and VCAM-1.Results:Contrasting the CD11b/CD18,ICAM-1 and VCAM-1 of the anoxic group and the group interfered with Chinese traditional medicine with chinese traditional medicine with the control group,we have found their expressions all more remarkably strengthened;There is some difference in expression between the anoxic group and the group interfered with Chinese traditional medicine,but it has little statistical significance.Conclusion:In this article,there occurs the mention:the chemotaxis of neutrophil has been strengthened and the endotheliocytosis of cerebrovascular has been conglutinated under anoxemia;as a result,the chemotaxis and endotheliocytosis have marred the brain cell tissue and participated in whole pathological and physiological process.

Effect of Adhesion Molecules on Skeletal Muscle Ischemia Reperfusion Injury / 中国康复理论与实践

@#Objective To investigate the changes of adhesion molecules and their effects on skeletal muscle ischemia/reperfusion injury. Methods 42 Wistar rats were divided into 3 groups: normal control group (Group Ⅰ, n=6), ischemia group (Group Ⅱ, n=6),ischemia/reperfusion injury group (Group Ⅲ, n=30). The level of malondialdehyde (MDA) in the plasma, myeloperoxidase (MPO) in the skeletal muscle, CD11b/CD18 on the leukocytes, intercelluar adhesion molecule-1 (ICAM-1) in the skeletal muscle and the histological changes were studied 1 h, 2 h, 4 h, 8 h, 12 h reperfusion after ischemia for 4 h. Results In group Ⅲ, the expression of CD11b/CD18, ICAM-1 and the injury of skeletal muscle increased with the lapse of reperfusion time. They reached the peak at 8～12 hours' reperfusion. The injury of skeletal muscle developed with the expression of adhesion molecules. Conclusion The expression of CD11b/CD18 and ICAM-1 are significantly associated with the skeletal muscle ischemia-reperfusion injury.

The Expression of CD 18 on Ischemia-Reperfusion Injury of TRAM Flap of Rats

PURPOSE: This study was to evaluate the expression pattern of CD 18(leukocyte adhesion glycoprotein) in ischemia-reperfusion injury of TRAM flap of rats. Through this study, we can obtain more information about ischemia-reperfusion injury. We want to develop specific medicine to improve the survival rate of TRAM flap in the future. METHODS: A TRAM flap supplied by a single pedicle superior epigastric artery and vein was elevated on 60 Sprauge-Dawley rats. The rats were divide into 6 groups (each group n=10); Group O: sham, no ischemia-reperfusion injury, Group I: 2 hour reperfusion after 4 hour ischemia, Group II: 4 hour reperfusion after 4 hour ischemia, Group III: 8 hour reperfusion after 4 hour ischemia, Group IV: 12 hour reperfusion after 4 hour ischemia, and Group V: 24 hour reperfusion after 4 hour ischemia. This study consisted of gross examination for flap survival and flow cytometry study of CD18 on neutrophils. RESULTS: The gross measurement of the flap showed different survival rate in group I(71%), II(68%), III(37%), IV(34%) and V(34%). All experimental groups showed an increase in the expression of CD18 compared to group O. The expression of CD18 was rapidly increased in ascending order in group I, II and III. But, the expression of CD18 was maintained in group IV and V. CONCLUSION: The results can be implemented in the study to develop drugs which are capable of reducing ischemia-reperfusion injury in microsurgical breast reconstruction.

Adhesion of CD40-stimulated Germinal Center B Cells to HK Cells Employs the CD11a/CD18-CD54 Interactions

BACKGROUND: The molecular basis of follicular dendritic cells (FDC)-germinal center (GC) B cell interaction is largely unknown, although this cellular interaction is thought to be important for the whole process of GC B cell differentiation. METHODS: Using FDC-like cells, HK, and highly purified GC B cells, we attempted to identify the molecules that play critical roles in the interactions between FDC and B cells. GC B cells were co-cultured with HK cells and soluble CD154 in the presence or absence of various function-blocking monoclonal antibodies to examine their effect on GC B cell binding to HK cells and B cell proliferation. RESULTS: Anti-CD11a and anti-CD54 antibodies inhibited GC B cell binding to HK cells while anti-CD49d and anti-CD106 antibodies did not. GC B cell proliferation was not impaired by the disruption of GC B cell-HK cell adherence. CONCLUSION: Our results suggest that CD11a/CD18-CD54 interactions play an important roles in the initial binding of GC B cells to FDC and diffusible growth factors from FDC may be responsible the massive proliferation of GC B cells.

Expression of adnesion molecules ICAM-1.CD11b/CD18 in chronic venous insufficiency and their clinical implication / 重庆医科大学学报

Objective:Explore the relationship between AMS expression and skin damage.Metheds:The study employed ELISA method,immunohistochemical method and electron minroscope analysis.Results:The results showed that the index of EC-ICAM-1 and PMN-CD11b plsitively expression increased remarkably in class 2~3 compared with that in class 1 and contrast group.The index of PMN-CD18 expression in class 2~3 and class 1 was greatly higher than that in contrast groups(P

The Effects of CD11c/CD18 and CD14 Blocking on Lipopolysaccharide-induced Endotoxemia

We studied the morphologic changes and the expression of cytokines of major organs by blocking CD14 and CD11c/CD18, which are known to be receptors of lipopolysaccharide (LPS), in the LPS induced endotoxemic mice. In 2 and 8 hours after initial intraperitoneal injection of 10 mg/kg of LPS into the mice, 500 microgram/kg of anti-CD14 antibody (Ab) and/or the same dosage of the anti-CD11c/CD18 Ab were administered intravenously, respectively or concomitantly. Under the light microscope, the LPS only and the LPS with the anti-CD14 Ab injected groups (group 1 and 2) showed marked acute inflammation in the organs, especially in the lung and liver, but the LPS with the anti-CD11c/CD18 Ab only or with both anti-CD14 and anti-CD11c Abs injected groups (group 3 and 4) revealed only mild acute inflammation. Under the electron microscope, there was marked inflammation in the group 1 and 2 such as marked infiltration of neutrophils, monocytes/ macrophages, lymphocytes, aggregation of platelets, and marked edematous change of endothelial cells with separation from the basement membrane. However in the group 3 and 4, there was only mild inflammation such as mild infiltration of neutrophils in the tissue or aggregation of neutrophils in the capillaries and sinusoids with mild endothelial swelling. The expressions of tumor necrosis factor-alpha (TNF-alpha) and interleukin-1alpha (IL-1alpha), detected by RT-PCR method, was remarkable in the group 2, but minimal in the group 3 and 4. We conclude that blocking the CD11c/CD18 with anti-CD11C/CD18 Ab is effective for the prohibition of biologic reactions and diminution of the inflammation induced by the LPS, even in the LPS induced endotoxemia.

The Effect of Cortisone on Generalized Shwartzman Reaction / 대한신장학회잡지

A generalized Shwartzman reaction(gSr) is characterized by vascular injury with microvascular fibrin deposition, leukocyte aggregation and hemorrhagic lesions in various organs and elicited by endotoxin from gram-negative bacteria. Leukocyte adherence to endothelial cells has been regarded as a pivotal event in this type of neutrophil mediated inflammatory response. Recently, it was described that leukocyte adhesion to endothelium is facilitated by the adhesion molecules, such as LFA-1(CDIIa/CD18) or Mac-1 (CDllb/CD18) binding to intercellular adhesion molecule-1(ICAM-1). To investigate the role of cell adhesion molecules in inflammatory vascular injury, the authors examined the expression of cell adhesion molecules, ICAM-1 and CD18, in gSr. Group A(n=5) received two injections of lipopolysaccharide(LPS) from E. coli 055:B5; 200 microgram/kg and 200 microgram/kg 24hours apart. Group B(n=5) received one dose of LPS(200 microgram/kg) after glucocorticoid treatment(methylprednisolone, 25mg/kg/day). Group C(n=5) received single dose of LPS(200 microgram/kg). The control group(n=3) received two injections saline instead of LPS. Animals were sacrificed four hours after the last dose of LPS or saline, The results were as follows: 1) Group A and B showed numerous fibrin micro-thrombi, inflammatory cell infiltration and some he-morrhagic change in kidney and lung, but group C and the control group did not show any lesions. In kidney the degree of fibrin deposition by IF was significantly increased in group A and B when compared to group C(p<0.05). 2) ICAM-1 was expressed by vascular endothelium including glomerular capillaries and some lymphocytes in the control group, This expression was increased in group C. But, group A and B was significantly decreased in ICAM-1 expression, especially in kidney comparing with group C and the control group(p<0.05). 3) Group A and B were significantly increased in CD18 expressing cells in glomerulus when compared with the control group(p<0.05). The number of CD18 expressing cells in glomerulus correlated significantly with a degree of fibrin deposition. In conclusion, these results suggest that the development of gSr requires the adhesion of activated inflammatory cells especially neutrophils to vascular endothelium and these are achieved by the interaction of adhesion molecules like ICAM-1 and CD18. Replacing the LPS with a glucocorticoid also expressing ICAM-1 on endothelium and prepares for leukocyte adhesion to endothelium.

Pretreatment of hypertonic saline attenuates the hepatic ischemia reperfusion injury induced by neutrophils / 中国病理生理杂志

AIM:To explore the effect of the pretreatment of hypertonic saline(HTS) in hepatic ischemia reperfusion(I/R) injury.METHODS:The rats were divided into sham group(sham group),ischemia reperfusion group(IR group) and pretreatment of hypertonic saline group(HTS group).Partial hepatic ischemia reperfusion model was used.The rats were sacrificed at the time of 1 h,3 h,6 h,12 h and 24 h after reperfusion in each group,respectively.Blood samples were obtained to examine ALT.The expression of the CD11b/CD18(Mac-1) on the neutrophils was analyzed by flow cytometry.RT-PCR and Western blotting were used to examine the expression of intercellular adhesion molecule-1(ICAM-1) in livers and chromatometry was performed to detect the activity of myeloperoxidase(MPO) in livers.The morphology of hepatocytes and the structure of sinusoid were observed by histological examinations.RESULTS:① HTS pretreatment decreased the level of ALT at the time points of 3 h,6 h and 12 h after reperfusion(P

Immunologic Changes of HL-60 Cells by Differentiation Inducing Agents I : Phenotypic Differentiation of FcgammaR and Mac-1

PURPOSE: Fc receptors and Mac-1 play an important role in the protective response of granulocytes and monocytes against microbial infection. FcgammaRl, FcgammaRll, FcgammaRlll as well as CD11b/ CD18 have never been measured in a quantitative way during hemopoiesis. Thus we quantified the expression of Fc Rl, Fc Rll, Fc Rlll, and CD11b/CD18 during hematopoietic differentiation using HL-60 cells, which was induced to differentiate by DMSO, or PMA. METHODS: HL-60 cells (ATCC CCL-240) were induced to differentiate by adding 1.0% DMSO, or 16nM PMA. On the 4th and 7th day after stimulation as well as before stimulation, phenotypic analysis was performed by flow cytometry after staining the cells with PE-conjugated anti- human CD64, CD32, CD16, CD11b, CD18, and isotype controls. And the measured fluorescent intensity was transformed into Molecules of Equivalent Soluble Fluorochromes (MESF). RESULTS: Percent positive cells and MESF of CD11b on HL-60 cells increased upon induction by DMSO, but not by PMA. Percent positive cells of CD18 on HL-60 cells was 99% regardless of differentiation. But MESF of CD18 was increased on the 4th day and decreased on the 7th day by DMSO or PMA. Percent positive cells and MESF of FcgammaRl on HL-60 cells increased upon induction by DMSO or PMA. Percent positive cells of FcgammaRll on HL-60 cells was above 90% regardless of differentiation. MESF of FcgammaRll showed no significant change by DMSO or PMA. CONCLUSION: Quantitative expression of FcgammaRl, FcgammaRll, FcgammaRlll, and CD11b/CD18 of HL-60 cells changed during induction of differentiation by DMSO or PMA. MESF of FcgammaR and CD11b/ CD18 a better indicator than percent positive cells to compare the differentiation of HL-60cells.

Prevention of reperfusion injury with CD18 monoclonal antibody and superoxide dismutase

Prolonged ischemia results in cellular necrosis and only prompt restoration of blood flow will prevent this type of injury. However, reperfusion itself can cause significant injury of previously ischemic tissue, i.e. "reperfusion injury'. This is an issue of concern in many areas of reconstructive surgery including free tissue transfer and replantation. Many factors have been implicated in the cause of reperfusion injury. Oxygen free radicals have enjoyed increasing popularity recently, but leukocytes had been thought to have a role only in the healing process that follows ischemic injury. Current studies in myocardium, liver and intestine have shown a dramatic increase in tissue leukocytes after ischemia-reperfusion and evidence implicating leukocytes in pathogenesis of ischemia-reperfusion injury has come from studies demonstrating significant injury reduction by depletion of circulating neutrophils. Therefore, increased neutrophil adhesiveness is a critical early step in the sequence of events leading to neutrophil-mediated injury. The purpose of this study is to evaluate the effect of CDl8 monoclonal antibody(CDl8 mAb), blocking antibody of neutrophil adherence, and superoxide dismutase (SOD), free radical scavenger, on reperfusion injury in rat epigastric island skin flap. The epigastric pedicle was occluded for six hours with ambient temperature at 22+/-1degrees C. The epigastric nerve was carefully dissected out and left intact to minimize autocannibalization. The flaps were sutured back down to their beds over interposed silicone sheets to prevent plasmatic imbibition. Fifteen minutes before reperfusion, the flaps were perfused with saline, CDl8 mAb(1 mg/kg), SOD(20,000 unit/kg) or CDl8 mAh/SOD(1 mg/kg + 20,000unit/kg). Percentage of flap survival was assessed by computerized planimetry on the seventh day. Tissue biopsies for myeloperoxidase(MPO) and malonyldialdehyde (MDA) were obtained at 24 hours after reperfusion. The results were as follows. 1. Percentage of flap survival was significantly increased in CDl8 mAb/SOD, CDl8 mAb and SOD groups in order, compared to the control(P < 0.05). Percentage of flap survival was significantly increased in CDl8 mAb group as compared with SOD group(p < 0.05). Percentage of flap survival significantly increased in CDl8 mAb/SOD group as compared with CDl8 mAb and SOD groups(p < 0.05) 2. MPO activity was significantly decreased in CDl8 mAb/SOD, CDl8 mAb and SOD groups(p < 0.01). MPO activity was significantly decreased in CDl8 mAb group as compared with SOD group. (p < 0.01). 3. MDA content was significantly decreased in CDl8 mAb/SOD, CDl8 mAb and SOD groups (p < 0.01), but the difference between CDl8 mAb and SOD groups was not significant. From those above results, we get to the conclusion that blocking neutrophil adherence and/or aggregation with monoclonal antibodies to CDl8 as compared with radical scavenger significantly ameliorates reperfusion injury. It is suggested that combination of modalities with antiadhesion therapy and radical scavenger may have a synergistic effect of improving flap survival and may be the optimal prevention of ischemiareperfusion injury.

Prevention of reperfusion injury with CD18 monoclonal antibody and superoxide dismutase

Prolonged ischemia results in cellular necrosis and only prompt restoration of blood flow will prevent this type of injury. However, reperfusion itself can cause significant injury of previously ischemic tissue, i.e. "reperfusion injury'. This is an issue of concern in many areas of reconstructive surgery including free tissue transfer and replantation. Many factors have been implicated in the cause of reperfusion injury. Oxygen free radicals have enjoyed increasing popularity recently, but leukocytes had been thought to have a role only in the healing process that follows ischemic injury. Current studies in myocardium, liver and intestine have shown a dramatic increase in tissue leukocytes after ischemia-reperfusion and evidence implicating leukocytes in pathogenesis of ischemia-reperfusion injury has come from studies demonstrating significant injury reduction by depletion of circulating neutrophils. Therefore, increased neutrophil adhesiveness is a critical early step in the sequence of events leading to neutrophil-mediated injury. The purpose of this study is to evaluate the effect of CDl8 monoclonal antibody(CDl8 mAb), blocking antibody of neutrophil adherence, and superoxide dismutase (SOD), free radical scavenger, on reperfusion injury in rat epigastric island skin flap. The epigastric pedicle was occluded for six hours with ambient temperature at 22+/-1degrees C. The epigastric nerve was carefully dissected out and left intact to minimize autocannibalization. The flaps were sutured back down to their beds over interposed silicone sheets to prevent plasmatic imbibition. Fifteen minutes before reperfusion, the flaps were perfused with saline, CDl8 mAb(1 mg/kg), SOD(20,000 unit/kg) or CDl8 mAh/SOD(1 mg/kg + 20,000unit/kg). Percentage of flap survival was assessed by computerized planimetry on the seventh day. Tissue biopsies for myeloperoxidase(MPO) and malonyldialdehyde (MDA) were obtained at 24 hours after reperfusion. The results were as follows. 1. Percentage of flap survival was significantly increased in CDl8 mAb/SOD, CDl8 mAb and SOD groups in order, compared to the control(P < 0.05). Percentage of flap survival was significantly increased in CDl8 mAb group as compared with SOD group(p < 0.05). Percentage of flap survival significantly increased in CDl8 mAb/SOD group as compared with CDl8 mAb and SOD groups(p < 0.05) 2. MPO activity was significantly decreased in CDl8 mAb/SOD, CDl8 mAb and SOD groups(p < 0.01). MPO activity was significantly decreased in CDl8 mAb group as compared with SOD group. (p < 0.01). 3. MDA content was significantly decreased in CDl8 mAb/SOD, CDl8 mAb and SOD groups (p < 0.01), but the difference between CDl8 mAb and SOD groups was not significant. From those above results, we get to the conclusion that blocking neutrophil adherence and/or aggregation with monoclonal antibodies to CDl8 as compared with radical scavenger significantly ameliorates reperfusion injury. It is suggested that combination of modalities with antiadhesion therapy and radical scavenger may have a synergistic effect of improving flap survival and may be the optimal prevention of ischemiareperfusion injury.