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AIM: To evaluate the efficacy of transplantation of human umbilical cord mesenchymal stem cells(hUCMSCs)in the treatment of corneal alkali burn in rabbits, and study the infiltration of polymorphonuclear neutrophils(PMNs)and the changes of vascular endothelial growth factor(VEGF)expression.METHODS: Corneal alkali burn models were established in right eyes of 75 healthy Japanese white rabbits, which were divided into three groups(group A, B and C), with 25 rabbits in each group. Group A was treated with amniotic membrane combined with hUCMSCs on the day after corneal alkali burn. Group B was treated with amniotic membrane only. Group C did not give any treatment after corneal alkali burn. At 3, 7, 14, 21 and 28d after corneal alkali burn, the corneal recovery was observed by slit lamp and photographed, the growth of corneal neovascularization(CNV)was scored, and corneal tissue was separated to make pathological sections. PMNs infiltration was observed by hematoxylin-eosin(HE)staining, and the expression of VEGF was determined by immunohistochemical staining.RESULTS: The growth of CNV in group A was much slower than that in group B at 14d after alkali burn. The CNV growth score around lesions of group A was significantly lower than that of group B(P<0.05). The quantity of PMNs increased on the 3d with the stromal layer of cornea infiltrated, relatively decreased on the 7d, shown a peak on the 14d, and then decreased gradually. Early infiltration after alkali burn was in the corneal stroma of the lesion area, and the extent of infiltration was equal to the ulcer area at later stage. The cell densities of corneal PMNs in group A and group B were significantly lower than those in group C at all time points after alkali burns(P<0.05), and those in group A were significantly lower than group B at 14 and 21d(P<0.05). The expression levels of corneal VEGF in all groups after alkali burn reached peak at 7～14d and decreased significantly at 28d, and the expression levels of VEGF in group A and group B at all time points after alkali burn were significantly lower than those in group C(P<0.05), and group A was significantly lower than that in group B at 7, 14 and 21d(P<0.05).CONCLUSION: The transplantation of hUCMSCs after alkali burn cornea can reduce the formation of CNV and inhibit corneal revascularization after alkali burn. The corneal pathological lesions and vascularization are closely related to PMNs and VEGF.
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Periodontitis is a chronic infectious disease leading to periodontal connective tissue destruction and alveolar bone resorption, which is widely prevalent and seriously endangers the oral and systemic health of a wide range of patients. The host immune inflammatory response plays a major role in the tissue destruction of periodontitis. Polymorphonuclear neutrophils (PMNs), as one of the important immune cell components in periodontal tissues, can trigger the host immune inflammatory response through the release of pro-inflammatory factors, which in turn leads to periodontitis. DNA methylation can influence the function of immune cells by regulating gene expression. Bioinformatics technology can provide new ideas for the treatment of periodontitis by analyzing the gene expression profiles and DNA methylation data of periodontal tissues from public databases of periodontitis patients and healthy populations, uncovering key DNA methylation genes of PMNs, and elucidating the influence of these genes on the pathological progression of periodontitis.
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@#AMI: To study the relationship between the infiltration of polymorphonuclear neutrophils(PMNs)and the expression of matrix metalloproteinases 9(MMP-9)in corneal stroma injury after alkali burn.<p>METHODS: Cornea alkali-burned model was made in 25 rabbits, then animals were grouped and sacrificed at 3d, 7d, 14d, 21d and 28d, respectively. The condition developing of alkali-burned cornea was observed by slit lamp biommicroscopy. The corneas were enucleated for histopathological examination. The infiltration of PMNs was identified by hematoxylin eosin(HE)staining and the expression of MMP-9 was identified by immunohistochemisty in different periods.<p>RESULTS: The quantity of PMNs and MMP-9 increased on the 3d, reached the lower level on 7d, shown a peak on the 14d, then decreased gradually. The area and depth of corneal stroma after alkali burn were the most severe on the 14d.<p>CONCLUSION: During the wound healing process, alkali-burned cornea has close relation with the infiltration of PMNs and the expression of MMP-9. The infiltration of PMNs and the expression of MMP-9 is positively correlated in corneal stroma injury after alkali burn.
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AIM: To study the infiltration of polymorphonuclear neutrophils ( PMNs ) after conjunctival flap covering in alkali-burned cornea. ●METHODS: Rabbit cornea alkali-burned model was made, then 50 rabbits were randomly divided into the experimental group ( n=25 ) and the control group ( n=25 ) . At the same time the surgery of conjunctival flap covering was given to rabbits of the experimental group. The condition developing of alkali-burned cornea was observed by slit lamp biomicroscopy, and took photos in two groups. The infiltration of PMNs was identified by hematoxylin eosin ( HE) staining in different periods. ●RESULTS:The quantity of PMNs increased on the 3d, reached the lower level on 7d, shown a peak on the 14d, then decreased gradually. PMNs level of the experimental group was significantly lower than that in the control group, and the difference of 3, 14 and 21d was significant (P ●CONCLUSION: During the wound healing process, alkali - burned cornea has close relation with the infiltration of PMNs. The treatment of conjunctival flap covering for the severe alkali-burned cornea was found to have good effect.
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Objective To investigate the effect of dexmedetomidine (Dex) on the apoptosis and respiratory burst of human polymorphonuclear neutrophils (PMN) in vitro. Methods The peripheral venous blood was collected from healthy volunteers. Isolation of PMN was performed by using the discontinuous plasma-Percoll gradient technique. PMN was randomLy divided into four groups: control group and three different concentrations of Dex (1 ng/mL,10 ng/mL,100 ng/mL) groups. PMN was cultured in enriched RPMI-1640 media at 2 × 106/mL for 24 h. The caspase-3 activity, apoptosis rate and respiratory burst of PMN were detected at 2 and 24 h. Results Compared with the control group, the PMN which was treated with three different concentrations of Dex showed no significant difference in caspase-3 activity, apoptosis rate and respiratory burst of PMN after 2 h of incubation. Compared with the control group, 1 ng/mL concentration of Dex did not affect the caspase-3 activity, apoptosis and respiratory burst of PMN cultured for 24 h. However, 10 ng/mL and 100 ng/mL concentrations of Dex promoted the caspase-3 activity, increased apoptosis rate and reduced the respiratory burst of PMN after 24 h of incubation. Compared with 10 ng/mL Dex group, 100 ng/mL concentrations of Dex promoted the caspase-3 activity, increased apoptosis rate and reduced the respiratory burst of PMN after 24 h of incutation. Conclusion Clinically relevant concentration of Dex does not affect apoptosis and respiratory burst of PMN, while high concentration of dexmedetomidine can induce apoptosis of PMN.
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Objective To study the different effects from different concentration ratios of polymorphonuclear neutrophil (PMN) to tumor cell (TC) on the process of tumor cell adhesion to endothelial cell (EC) in shear flow. Methods PMNs and TCs with different concentration ratios (PMN-TC ratio) were added into the parallel plate flow chamber, and changes in the numbers of transient and accumulative adhered TCs on ECs at different shear rates (50 s-1，100 s-1，200 s-1) were analyzed. Results The transient and accumulative adhesion of TCs on ECs at PMN-TC ratio of 3︰1 significantly increased as compared to that at PMN-TC ratio of 1︰1, especially under high shear flow condition (100 s-1 and 200 s-1). Moreover, in the 5 minute-observation period, the effect of PMN-TC ratio on TC adhered to ECs occurred earlier when the shear rate increased. Conclusions The increase of PMN-TC concentration ratio can promote TC adhesion to ECs in shear flow, and the research findings provide significant references for studying TC metastasis in blood vessels and the target therapy of tumors.
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BACKGROUND: Cardiopulmonary bypass (CPB) induces variable systemic inflammatory reactions associated with major organ dysfunction via polymorphonuclear neutrophils (PMNs). Ulinastatin, a urinary trypsin inhibitor, inhibits PMN activity and reduces systemic inflammatory responses. The aim of this study is to evaluate the effect of ulinastatin on postoperative blood loss and laboratory changes in patients undergoing open heart surgery. MATERIALS AND METHODS: Between January 2008 and February 2009, 110 patients who underwent atrioventricular valve surgery through right thoracotomy were divided into two groups. Patients received either 5,000 U/kg ulinastatin (ulinastatin group, n=41) or the equivalent volume of normal saline (control group, n=69) before aortic cross clamping. The primary end points were early coagulation profile changes, postoperative blood loss, transfusion requirements, and duration of intubation and intensive care unit stay. RESULTS: There were no statistically significant differences between the two groups in early coagulation profile, other perioperative laboratory data, and postoperative blood loss with transfusion requirements. CONCLUSION: Administration of ulinastatin during operation did not improve the early coagulation profile, postoperative blood loss, or transfusion requirements of patients undergoing open heart surgery. In addition, no significant effect of ulinastatin was observed in major organs dysfunction, systemic inflammatory reactions, or other postoperative profiles.
Subject(s)
Humans , Cardiopulmonary Bypass , Constriction , Glycoproteins , Heart , Hemostasis , Intensive Care Units , Intubation , Neutrophils , Postoperative Hemorrhage , Thoracic Surgery , Thoracotomy , TrypsinABSTRACT
BACKGROUND: Cardiopulmonary bypass (CPB) induces variable systemic inflammatory reactions associated with major organ dysfunction via polymorphonuclear neutrophils (PMNs). Ulinastatin, a urinary trypsin inhibitor, inhibits PMN activity and reduces systemic inflammatory responses. The aim of this study is to evaluate the effect of ulinastatin on postoperative blood loss and laboratory changes in patients undergoing open heart surgery. MATERIALS AND METHODS: Between January 2008 and February 2009, 110 patients who underwent atrioventricular valve surgery through right thoracotomy were divided into two groups. Patients received either 5,000 U/kg ulinastatin (ulinastatin group, n=41) or the equivalent volume of normal saline (control group, n=69) before aortic cross clamping. The primary end points were early coagulation profile changes, postoperative blood loss, transfusion requirements, and duration of intubation and intensive care unit stay. RESULTS: There were no statistically significant differences between the two groups in early coagulation profile, other perioperative laboratory data, and postoperative blood loss with transfusion requirements. CONCLUSION: Administration of ulinastatin during operation did not improve the early coagulation profile, postoperative blood loss, or transfusion requirements of patients undergoing open heart surgery. In addition, no significant effect of ulinastatin was observed in major organs dysfunction, systemic inflammatory reactions, or other postoperative profiles.
Subject(s)
Humans , Cardiopulmonary Bypass , Constriction , Glycoproteins , Heart , Hemostasis , Intensive Care Units , Intubation , Neutrophils , Postoperative Hemorrhage , Thoracic Surgery , Thoracotomy , TrypsinABSTRACT
Objective To explore the impact of levels of serum IL-17,Leukotriene B4 and IgE on pathogenesis of childhood asthma.Methods Totally 60 children with asthma acute exacerbation ( 29 children with mild asthma,31 children with moderate-severe asthma) were selected as study group,24 healthy children were selected as control group.Serum IL-17 and LTB4 were measured with euryzemLinked immunosorbent assay,serum IgE was determined with enzyme-linked fluoroimmuneassay by pharmacia CAP Sytem,PMN was determined with automatic blood analyser,pulmonary function was measured in the study group.Results ( 1 ) The level of serum IL-17 ( 1.15 ± 0.10 μg/L,2.80 ± 2.30 μg/L,0.83 ± 0.10 μg/L),LTB4 (2.22 ± 1.01 μg/L,8.79 ± 9.36 μg/L,1.94 ± 1.13 μg/L) and IgE( 123.70 ±86.94 μg/L,322.27 ±332.28 μg/L,24.27 ±7.64 μg/L) were significantly different among mild asthma group,moderate-severe asthma group and control group( P ＜ 0.001 ).( 2 )The N％ of mild asthma group,moderate-severe asthma group and study group were( 55.06 ± 1 1.15 ) ％,( 64.44± 11.87)％,(47.96 ± 13.52)％,L％ were(42.20 ± 11.04)％,(33.93 ± 10.02)％,(49.65 ± 13.02)％,and there were significant differences in N％ and L％ between study group and control group( P ＜ 0.05 ).( 3 ) There were significant positive correlations between the serum IL-17 levels and IgE,LTB4 and IgE,IL-17 and LTB4 in asthmatic children( P ＜0.05).(4) There were significant negative correlcations between the level of serum IL-17,LTB4 and FEVI,PEF( P ＜0.001 ).There were significant positive correlations between serum IL-17,LTB4 and N％ (P ＜0.001 ).(5)There were not correlations between the level of serum IgE and FEV1,PEF and N％in asthmatic children( P ＞0.05 ).Conclusion The levels of serum IL-17,LTB4 and IgE participated in pathogenesis on asthmatic children patients.
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Recent studies have reported that exogenous gangliosides, the sialic acid-containing glycosphingolipids, are able to modulate many cellular functions. We examined the effect of micelles of mono- and trisialoganglioside GM1 and GT1b on the production of reactive oxygen species by stimulated human polymorphonuclear neutrophils using different spectroscopic methods. The results indicated that exogenous gangliosides did not influence extracellular superoxide anion (O2.-) generation by polymorphonuclear neutrophils activated by receptor-dependent formyl-methionyl-leucyl-phenylalanine. However, when neutrophils were stimulated by receptor-bypassing phorbol 12-myristate 13-acetate (PMA), gangliosides above their critical micellar concentrations prolonged the lag time preceding the production in a concentration-dependent way, without affecting total extracellular O2.- generation detected by superoxide dismutase-inhibitable cytochrome c reduction. The effect of ganglioside GT1b (100 µM) on the increase in lag time was shown to be significant by means of both superoxide dismutase-inhibitable cytochrome c reduction assay and electron paramagnetic resonance spectroscopy (P < 0.0001 and P < 0.005, respectively). The observed phenomena can be attributed to the ability of ganglioside micelles attached to the cell surface to slow down PMA uptake, thus increasing the diffusion barrier and consequently delaying membrane events responsible for PMA-stimulated O2.- production.
Subject(s)
Humans , G(M1) Ganglioside/pharmacology , Gangliosides/pharmacology , Neutrophils/drug effects , Reactive Oxygen Species/metabolism , Superoxide Dismutase/biosynthesis , Cytochromes c/pharmacology , Electron Spin Resonance Spectroscopy , Micelles , Neutrophils/metabolismABSTRACT
BACKGROUND: Ischemic preconditioning (IPC) and propofol has been demonstrated to inhibit postischemic adhesion of PMNs on coronary vascular endothelium, respectively. The purpose of this study is to investigate whether IPC and propofol inhibit postischemic coronary vascular adhesion of PMNs synergistically. METHODS: Twenty-eight isolated rat hearts were subjected to stabilization for 15 min, perfusion for 15 min, global ischemia for 15 min and reperfusion for 60 min serially. The isolated hearts were divided into 4 groups as follows: control group, PMNs were infused for 1 min on 1 min after reperfusion following global ischemia; IPC group, IPC was performed before global ischemia and PMNs were infused as the same manner; PPF group, Propofol (0.3microgram/ml) was infused from perfusion until 5 min after reperfusion and PMNs were infused as the same manner; IPC/PPF group, IPC was performed before global ischemia and propofol was infused from perfusion until 5 min after reperfusion and PMNs were infused as the same manner. PMNs adhesion, heart rate, left ventricular developed pressure, dP/dt, coronary flow and creatinine kinase enzyme were measured. RESULTS: Postischemic PMNs adhesion was reduced in IPC, propofol, and IPC/PPF group compared to the control group (30.8 +/- 7.3%, 32.0 +/- 3.9%, 20.9 +/- 6.1% vs 75.7 +/- 6.3%, P <0.05, respectively). There were no significant differences in PMNs postischemic adhesion between IPC group and PPF group, respectively and IPC/PPF group. There were no significant differences in changes in HR, LVDP, dP/dt, CF and CK among the four group. CONCLUSIONS: These results suggest that IPC and propofol have no synergistic effect on reducing postischemic adhesion of PMNs.
Subject(s)
Animals , Rats , Creatinine , Endothelium, Vascular , Heart , Heart Rate , Ischemia , Ischemic Preconditioning , Neutrophils , Perfusion , Phosphotransferases , Propofol , Reperfusion , Reperfusion InjuryABSTRACT
BACKGROUND: Adhesion of polymorphonuclear neutrophils (PMNs) to the coronary vascular endothelium is a crucial step in the development of postischemic myocardial injury. Propofol, a free radical scavenger has been demonstrated to provide cardioprotective effects by attenuating ischemia and reperfusion injury. The purpose of this study was to investigate whether propofol inhibits the postischemic coronary vascular adhesion of PMNs and results in a reduction of postischemic myocardial dysfunction in isolated guinea pig hearts. METHODS: 42 male guinea pigs were used in this study. Hearts were isolated and perfused with modified Krebs-Hanseleit solution. All isolated hearts were subjected in turn to stabilization for 10 min, perfusion for 15 min, global ischemia for 30 min and reperfusion for 60 min. The isolated hearts were then divided into 6 groups. Each group was subjected to different interventions as follow so. C group: No intervention was performed, except ischemia/reperfusion injury. PPF group: Propofol (4 microgram/ml) was infused from the start of perfusion to 5 min after the start of reperfusion. PMN group: PMNs were infused for 1 min on 2 minutes after reperfusion. P2PMN group: Propofol (2 microgram/ml) was infused from the start of perfusion to 5 min after the start of reperfusion, and PMNs were infused for 1 min on 2 minutes after reperfusion. P4PMN group: Propofol (4 microgram/ml) was infused from the start of perfusion to 5 min after the start of reperfusion, and PMNs were infused for 1 min on 2 minutes after reperfusion. IL-PMN group: Intralipid (0.4 mg/ml) was infused from the start of perfusion to 5 min after the start of reperfusion, and PMNs were infused for 1 min on 2 minutes after reperfusion. PMNs adhesion (%), left ventricular developed pressure (LVDP), dP/dt, heart rates and coronary flow were measured. RESULTS: Propofol (2 microgram/ml, 4 microgram/ml) significantly reduced postischemic PMNs adhesion compared with that of the PMN group (54.0+/-8.0%, 50.0+/-5.0% vs 72.5+/-6.5% P <0.05 respectively) and prevented the deterioration of myocardial function seen after PMNs application (LVDP 74.5+/-7.3%, 60.3+/-4.5% vs 48.4+/-2.7% respectively). CONCLUSIONS: These results show that propofol reduces the postischemic coronary vascular adhesion of PMNs and prevents postischemic myocardial dysfuncion.
Subject(s)
Animals , Humans , Male , Endothelium, Vascular , Guinea Pigs , Heart , Heart Rate , Ischemia , Neutrophils , Perfusion , Propofol , Reperfusion , Reperfusion InjuryABSTRACT
BACKGROUND: The adhesion of polymorphonuclear neutrophils (PMNs) to the coronary endothelium is an crucial step in PMN-mediated reperfusion injury. The purpose of this study was to investigate whether thiopental inhibits the postischemic coronary vascular adhesion of PMNs, and results in reduced postischemic myocardial dysfunction in isolated guinea pig hearts. METHODS: Hearts (n = 6-8/group) were isolated from male-guinea pigs and perfused with modified Krebs-Henseleit solution. After isolation, hearts were stabilized for 10 minutes, perfused for 15 minutes, allowed an ischemic period for 30 minutes, and a reperfusion period of 60 minutes (C group). In the P group, a bolus of 1x10(6) PMNs was infused 2 minutes after reperfusion, and in the T group additional thiopental was infused 5 minutes after the start of reperfusion, and PMNs were infused on 2 minutes after reperfusion. PMN adhesion (%), LVDP, LVEDP, +/-dP/dt, HR, and CF were measured pre- and postischemia. RESULTS: The addition of thiopental (25 microM) to the perfusate reduced the postischemic coronary vascular adhesion of PMNs (72.5+/-4.5% vs 40.0+/-7.4%, P < 0.05) and prevented postischemic myocardial dysfunction compared with group P (73.5+/-6.9% vs 48.4+/-3.0%, P < 0.05). CONCLUSIONS: Postischemic myocardial dysfunction is significantly more pronounced after PMN infusion. Thiopental reduced the postischemic coronary vascular adhesion of PMNs and attenuated the myocardial dysfunction, which was responsible at least in part, for the cardioprotective effect.
Subject(s)
Animals , Endothelium , Guinea Pigs , Guinea , Heart , Ischemia , Neutrophils , Reperfusion , Reperfusion Injury , Swine , ThiopentalABSTRACT
Objective To investigate the effects of polysaccharide sulfate(PSS)on the growth of vascular endothelial cell cultured in vitro and the adhesion of vascular endothelial cell to polymorphonuclear neutrophils(PMN).Methods ECV-304,a kind of human umbilical vein endothelial cells,was cultured in vitro with different concentrations of PSS(0,50,100,150,200,250 g?mL-1),the growth of ECV-304 was observed by means of MTT.Hypoxia/ reoxygenation(H/R) model of ECV-304 was established,the adhesion rate of PMN from patients with cerebral infarction to cultured ECV-304 was detected with or without treatment of PSS and H/R.Results The growth of ECV-304 in PSS-treated subgroups(final concentrations: 50,100,150,200 g?mL-1) were much better than in non-PSS subgroup,especially in subgroup with PSS final concentration of 100 g?mL-1.The adhesion rate of cultured ECV-304 to PMN from patients with cerebral infarction was higher significantly than that from the healthy subject,which was increased when ECV-304 being treated with H/R.The adhesion rate of cultured ECV-304(whether treated with H/R or not) to PMN was decreased significantly when ECV-304 being treated with PSS(100 g?mL-1).Conclusion The experiments in vitro showed that PSS could promote the growth of cultured ECV-304 and decrease the adhesion rate of H/R ECV-304 to PMN.