【Abstract】 Objective To discuss the role of leukocyte activation and inflammatory processes in the disease of chronic venous insufficiency （CVI）. Methods The relevant literatures about the role of leukocyte activation and inflammatory reaction in CVI were reviewed. Results The role of inflammatory reaction in occurrence and development of venous diseases has been studied a lot in recent years. It was found that the leukocyte activation and inflammatory reaction are involved in the structural remodeling of venous valves and walls, leading to valvular incompetence and formation of varicose veins. Conclusion Leukocyte activation and inflammatory processes take important roles in the occurrence and progression of CVI.
ObjectiveTo observe the adhension and stracking of leukocyte in the capillary vessels, and investigate the relationship between leukocyte and microvascular morphologic changes in retinal microvesselsof rats with early diabetes.MethodsA total of 90 healthy adult male Wistar rats were randomly divided into control and diabetes (induced by Streptozotocin, STZ) groups with 45 rats in each group. The rats in the diabetic group were further divided into 3, 7, and 14 days groups with 5 rats in each group, and 30, 90, and 180 days groups with 10 rats in each group. The right eyes of rats in each group were prepared for retinal digest preparations. The expression of leukocyte common antigen (CD45) was detected by immunohistochemical staining.ResultsFew CD45 positive cells in the retinal capillaries were seen in the control group. The expression of CD45 was significantly increased in the retinal capillaries 3 days after diabetes induction, and reached a peak at the 14th day. Morphological changes including capillary telangiectasia, atresia, and irregularity of capillary caliber were found in the retinal capillaries of rats 90 days after diabetes induction. The changes were aggravated 180 days after diabetes induction.ConclusionLeukocyte adhesion occurs in the early stage of diabetic retinopathy (DR), and is the beginning of the microvascular pathological changes. Leukocyte adhesion may play an important role in the occurrence and development of DR as the foundation of microvascular morphological changes.(Chin J Ocul Fundus Dis, 2003,19:344-347)
Objective To calculate the recovery rate and enrichment factor and to analyse the correlation by measuring the concentrations of platelets, leukocyte, and growth factors in platelet-rich plasma (PRP) so as to evaluate the feasibil ity and stabil ity of a set of PRP preparation. Methods The peripheral blood (40 mL) was collected from 30 volunteers accorded with the inclusion criteria, and then 4 mL PRP was prepared using the package produced by Shandong Weigao Group Medical Polymer Company Limited. Automatic hematology analyzer was used to count the concentrations of platelets and leukocyte in whole blood and PRP. The enrichment factor and recovery rate of platelets or leukocyte were calculated; the platelet and leukocyte concentrations of male and female volunteers were measured, respectively. The concentrations of platelet-derived growth factor (PDGF), transforming growth factor β (TGF-β), and vascular endothel ial growth factor (VEGF) were assayed by ELISA. Results The platelet concentrations of whole blood and PRP were (131.40 ± 29.44) × 109/L and (819.47 ± 136.32) × 109/L, respectively, showing significant difference (t=—27.020, P=0.000). The recovery rate of platelets was 60.85% ± 8.97%, and the enrichment factor was 6.40 ± 1.06. The leukocyte concentrations of whole blood and PRP were (5.57 ± 1.91) × 1012/L and (32.20 ± 10.42) × 1012/L, respectively, showing significant difference (t=—13.780, P=0.000). The recovery rate of leukocyte was 58.30% ± 19.24%, and the enrichment factor was 6.10 ± 1.93. The concentrations of platelets and leukocyte in PRP were positively correlated with the platelet concentration (r=0.652, P=0.000) and leukocyte concentration (r=0.460, P=0.011) in whole blood. The concentrations of platelet and leukocyte in PRP between male and female were not significantly different (P gt; 0.05). The concentrations of PDGF, TGF-β, and VEGF in PRP were (698.15 ± 64.48), (681.36 ± 65.90), and (1 071.55 ± 106.04) ng/ mL,which were (5.67 ± 1.18), (6.99 ± 0.61), and (5.74 ± 0.83) times higher than those in the whole blood, respectively. PDGF concentration (r=0.832, P=0.020), TGF-β concentration (r=0.835, P=0.019), and VEGF concentration (r=0.824, P=0.023) in PRP were positively correlated with platelet concentration of PRP. Conclusion PRP with high concentrations of platelets, white blood cells and growth factors can be prepared stably by this package.
Objective To review the treatment methods and techniques of ischemia-reperfusion injury of flap. Methods Recent basic research l iterature concerning ischemia-reperfusion injury of flap was reviewed and analyzed in terms of treatment techniques. Results Ischemia-reperfusion injury is one of the leading causes of flap necrosis postoperatively. Interventions against any l ink of the ischemia-reperfusion injury progress could effectively reduce the damageand improve the survival rate of flaps. Conclusion Including production of reactive oxygen species, neutrophil infiltrationetc are thought to be the main mechanisms of ischemia-reperfusion injury. Treatment including medicine administration and physical intervention against any specific l ink of ischemia-reperfusion injury can interfere or block the whole progress, which reduce the damage of ischemia-reperfusion injury and improve the survival rate of animal flap models eventually.
ObjectiveTo evaluate the association between extent and severity of acute coronary syndrome and uric acid, leukocytes. MethodsA retrospective analysis of leukocytes, platelets, lipids and uric acid levels were performed on 23 patients with acute myocardial infarction (AMI group), 17 patients with unstable angina (UA group), and 17 healthy subjects (controls) between January and December 2010. ResultsIn the three groups (AMI, UA, and Control), the leukocyte count was respectively (10.4±3.2)×109/L, (6.9±2.4)×109/L and (5.4±1.1)×109/L (P<0.05); neutrophil was (7.4±3.2)×109/L, (4.8±2.3)×109/L, and (3.4±0.8)×109/L (P<0.001); and uric acid was (401.4±94.3), (384.1±74.1) and (285.5±76.8) μmol/L, respectively (P<0.001). Multinomial Logistic regression showed leukocyte was a predictor for AMI (OR=1.712, P=0.003), while uric acid was not (OR=1.006, P=0.255), regarding the UA group as the reference. When the control group was using as reference, leukocyte was an independently significant factor for AMI (OR=2.942, P=0.004) and was not a significant factor for UA (OR=1.718, P=0.125); uric acid was a significant factor for AMI and UA (OR=1.027, P=0.016; OR=1.021, P=0.041). ConclusionUric acid may be associated with the chronic development of coronary heart disease, while leukocytes may play a potential role in plaque destabilization and the onset of AMI.
Objective To compare effectiveness of injecting platelet-rich plasma (PRP) with different concentrations of leukocytes under ultrasound in treatment of supraspinatus tendon injury. Methods A clinical data of 30 patients with supraspinatus tendon injury, who met the selection criteria and were admitted between December 2022 and December 2023, was retrospectively analyzed. Thirty patients were treated with 4 injections of leukocyte-poor PRP (LP-PRP, n=10), leukocyte-rich PRP (LR-PRP, n=11), and triamcinolone (n=9), with an interval of 7-10 days between each injection. There was no significant difference between groups (P>0.05) in the age, gender, disease duration, affected shoulder side, Ellman classification, preoperative visual analogue scale (VAS) score, Constant score, Disabilities of the Arm, Shoulder, and Hand (DASH) score, and American Shoulder and Elbow Surgeons (ASES) score. At 1, 3, and 6 months after injection, the shoulder pain and function were evaluated by using the VAS score, Constant score, DASH score, and ASES score. MRI was conducted to observe supraspinatus tendon healing. Results No severe adverse reactions was observed in all groups. All patients were followed up 6-7 months, with an average of 6.5 months. After injection, the ASES score and Constant score gradually increased in the LR-PRP group and LP-PRP group, while the VAS score and DASH score decreased, with significant differences compared to before injection (P<0.05). Except for no significant difference between 3 and 6 months after injection in LR-PRP group (P>0.05), the above scores showed significant differences between different time points (P<0.05). At 1 month after injection, the Constant score in triamcinolone group significantly increased compared to before injection, while the VAS score significantly decreased (P<0.05). There was no significant difference in all scores between other time points in the triamcinolone group (P>0.05). Except for 1 month after injection, there was no significant difference in Constant score and VAS score between groups (P>0.05). At all other time points, the LR-PRP group and LP-PRP group had better scores than the triamcinolone group (P<0.05). There was no significant difference between the LR-PRP group and the LP-PRP group (P>0.05). MRI showed that only 4 patients in the LP-PRP group had signs of repair at the supraspinatus tendon injury site at 6 months after injection, while no significant tendon repair sign was observed in the other patients. Conclusion Compared with triamcinolone treatment, multiple injections of LP-PRP and LR-PRP under ultrasound can promote the recovery of shoulder joint function and significantly relieve pain in patients with supraspinatus tendon injury, and imaging improvement can be seen after LP-PRP treatment.
Objective To assess the protective effects of a new type of leukocyte-depletion filter-1 (LD-1) on red blood cells during cardiopulmonary bypass(CPB). Methods Twelve Mongolian dogs, weight range 25-30kg, were divided into control group and leukocyte depletion group (LD group) with random number table, LD group (n=6) had our new type of leukocyte depletion filter-1 placed in venous line which was used within the first 5 minutes after onset of CPB. The control group (n=6) had no leukocyte depletion filter installed in the circuit. CPB was set up by cannulated with a venous cannula through the right atrium and with an aortic cannula after median sternotomy. Aorta was clamped at 10 minutes of CPB and released at 70 minutes of CPB. Dogs were observed for 2 hours after weaning from CPB. Blood samples were collected prior to, at 10, 40, 75 minutes, end of and 2 hours after CPB to determine circulating leukocytes, erythrocyte fragility and plasma levels of malondialdehyde(MDA), superoxide dismutase(SOD) and free hemoglobin(FHB). Results Leukocyte numbers were significantly reduced in LD group during CPB(Plt;0.01), and lower than those in control group (Plt;0.05). Plasma levels of SOD dropped after 75 minutes of CPB in control group, but those kept normal in LD group, and higher than those in control group at 2 hours after CPB (Plt;0.05, 0.01). Serum MDA and FHB levels increased sharply in two groups (Plt;0.01), but were lower in LD group than those in control group. The concentrations of NaCl when starting and complete hemolysis were also lower in LD group than those in control group at end of and 2 hours after CPB. Conclusion The new type of LD-1 used in venous line only 5 minutes after onset of CPB can decrease leukocyte counts, and reduce erythrocyte injury effectively.
ObjectiveTo observe the effects of p21 activated kinase 4 (PAK4) on the mitochondrial function and biological behavior in retinal vascular endothelial cells. MethodsThe experimental study was divided into two parts: in vivo animal experiment and in vitro cell experiment. In vivo animal experiments: 12 healthy C57BL/6J male mice were randomly divided into normal control group and diabetes group, with 6 mice in each group. Diabetes mice were induced by streptozotocin to establish diabetes model. Eight weeks after modeling, quantitative real-time polymerase chain reaction and Western blots were performed to detect the expression of PAK4 in diabetic retinas. In vitro cell experiments: the human retinal microvascular endothelial cells (hRMEC) were divided into three groups: conventional cultured cells group (N group), empty vector transfected (Vector group); pcDNA-PAK4 eukaryotic expression plasmid transfected group (PAK4 group). WB and qPCR were used to detect transfection efficiency, while scratching assay, cell scratch test was used to detect cell migration in hRMEC of each group. In vitro white blood cell adhesion experiment combined with 4 ', 6-diamino-2-phenylindole staining was used to detect the number of white blood cells adhering to hRMEC in each group. The Seahorse XFe96 cell energy metabolism analyzer measures intracellular mitochondrial basal respiration, adenosine triphosphate (ATP) production, maximum respiration, and reserve respiration capacity. The t-test was used for comparison between the two groups. Single factor analysis of variance was used for comparison among the three groups. ResultsIn vivo animal experiments: compared with normal control group, the relative expression levels of PAK4 mRNA and protein in retina of diabetic mice were significantly increased, with statistical significance (t=25.372, 22.419, 25.372; P＜0.05). In vitro cell experiment: compared with the N group and Vector group, the PAK4 protein, mRNA relative expression and cell mobility in the hRMEC of PAK4 group were significantly increased, with statistical significance (F=36.821, 38.692, 29.421; P＜0.05). Flow cytometry showed that the adhesion number of leukocytes on hRMEC in PAK4 group was significantly increased, and the difference was statistically significant (F=39.649, P＜0.01). Mitochondrial pressure measurement results showed that the capacity of mitochondrial basic respiration, ATP production, maximum respiration and reserve respiration in hRMEC in PAK4 group was significantly decreased, with statistical significance (F=27.472, 22.315, 31.147, 27.472; P＜0.05). ConclusionOver-expression of PAK4 impairs mitochondrial function and significantly promotes leukocyte adhesion and migration in retinal vascular endothelial cells.
The increased morbidity and mortality following cardiopulmonary bypass （CPB） may be due to the development of systemic inflammatory response syndrome （SIRS）. Leukocyte, especially neutrophil, plays a crucial role in SIRS during and after CPB, so the leukocyte removal by filtrations appears to be a logical anti-inflammatory strategy. Many articles reported that leukocyte depletion filter can decrease the potential adverse effects during CPB and reduce the morbidity and mortality following CPB. But the protective effects of the filter varied greatly from paper to paper. This may be due to the different design and biocompatibility of the filter at present, But, because the leukocyte plays a central role in SIRS, leukocyte depletion filter would be an important apparatus in therapy of noninfectious inflammation induced by CPB after it was improved.
ObjectiveTo identify a more popularized preparation protocol of leukocytes-rich platelet-rich plasma (L-PRP) for higher tolerance rate.MethodsThe peripheral blood samples of 76 volunteers (45.0 mL/case) were mixed with 5 mL sodium citrate injection for blood transfusion, and L-PRP was prepared by twice centrifugations. All blood samples were divided into three groups according to the parameters of twice centrifugation: experimental group A (12 cases, 400×g, 10 minutes for the first time and 1 100×g, 10 minutes for the second time), experimental group B (27 cases, 800×g, 10 minutes for the first time and 1 100×g, 10 minutes for the second time), and control group (37 cases, 1 360×g, 10 minutes for the first time and 1 360×g, 10 minutes for the second time). The platelet recovery rate and platelet and leukocyte enrichment coefficient of L-PRP in each group were calculated and compared.ResultsAfter removal of abnormal blood samples (platelet recovery rate was more than 100% or white thrombus), the remaining 55 cases were included in the statistical analysis, including 10 cases in experimental group A, 21 cases in experimental group B, and 24 cases in control group. The platelet enrichment coefficient and platelet recovery rate of experimental group B were significantly higher than those of experimental group A and control group (P<0.05); there was no significant difference between experimental group A and control group (P>0.05). There was no significant difference in leukocyte enrichment coefficient between experimental groups A, B, and control group (P>0.05).ConclusionThe preparation quality of PRP is affected by various factors, including centrifugal force, centrifugal time, temperature, and operation process, etc. Twice centrifugation (800×g, 10 minutes for the first time and 1 100×g, 10 minutes for the second time) is an ideal and feasible centrifugation scheme, which can obtain satisfactory platelet recovery rate and enrichment coefficient with thicker buffy coat, which can reduce the fine operation requirements for operators, improve the fault tolerance rate and generalization.