AIM: To evaluate the diagnostic capability of calprotectin in ascitic fluid for detecting a polymorphonuclear (PMN) cell count > 250/μL ascites. METHODS: In this prospective observational study, a total of 130 asc...AIM: To evaluate the diagnostic capability of calprotectin in ascitic fluid for detecting a polymorphonuclear (PMN) cell count > 250/μL ascites. METHODS: In this prospective observational study, a total of 130 ascites samples were analysed from 71 consecutive patients referred for paracentesis. Total and differential leukocyte cell counts were determined manually with a Neubauer chamber and gentianviolet stain. Calprotectin was measured in 1 mL ascetic fluid by enzyme-linked immunosorbent assay (ELISA) and a point-of-care (POC) lateral flow assay with the Quantum Blue Reader (Bühlmann Laboratories). All measurements were carried out in a central laboratory by senior personnel blinded to patient history. A PMN count > 250/μL was the primary endpoint of the study. The diagnostic value of ascitic calprotectin measurement was assessed by comparing to the final diagnosis of each patient that had been adjudicated by investigators blinded to calprotectin values. RESULTS: The PMN count was > 250/μL in 19 samples (14.6%) from 15 patients (21.1%) and varied widely among the study population (range 10-19 800/mL and 1-17 820/mL, respectively). Spontaneous bacterial peritonitis (SBP) was the final diagnosis in four patients (5.6%). All patients with PMN ≤ 250/μL had negative bacterial culture. PMN count was elevated in five patients with peritoneal carcinomatosis, three with lymphoma, one with neuroendocrine carcinoma, and two with secondary peritonitis due to abdominal perforation. PMN cell counts correlated with ascitic calprotectin values (Spearman's rho; r = 0.457 for ELISA, r = 0.473 for POC). A considerable range of ascitic calprotectin concentrations was detected by ELISA [median 0.43 μg/mL, interquartile range (IQR) 0.23-1.23 (range 0.10-14.93)] and POC [median 0.38 μg/mL, IQR 0.38-0.56 (range 0.38-13.31)]. Ascitic calprotectin levels were higher in samples with PMN > 250/μL, by both ELISA [median (IQR) 2.48 μg/mL (1.61-3.65) vs 0.10 μg/mL (0.10-0.36), P < 0.001] and POC [2.78 μg/mL (2.05-5.37) vs 0.38 μg/mL (0.38-0.41), P < 0.001]. The area under the receiver operating characteristics curve for identifying an elevated PMN count was 0.977 (95%CI: 0.933 to 0.995) for ELISA and 0.982 (95%CI: 0.942 to 0.997) for POC (P = 0.246 vs ELISA). Using the optimal cut-off value for ELISA (0.63 μg/mL), ascitic calprotectin had 94.8% sensitivity, 89.2% specificity, positive and negative likelihood ratios of 8.76 and 0.06 respectively, positive and negative predictive values of 60.0% and 99.0% respectively, and 90.0% overall accuracy. Using the optimal cut-off value for POC (0.51 μg/mL), the respective values were 100.0%, 84.7%, 6.53, 0.00, 52.8%, 100% and 87.7%. Correlation between ELISA and POC was excellent (r = 0.873, P < 0.001). The mean ± SD of the difference was -0.11 ± 0.48 μg/mL with limits of agreement of + 0.8 μg/mL (95%CI: 0.69 to 0.98) and -1.1 μg/mL (95%CI: -1.19 to -0.91). CONCLUSION: Ascitic calprotectin reliably predicts PMN count > 250/μL, which may prove useful in the diagnosis of SBP, especially with a readily available bedside testing device.展开更多
The plasma effects during hypoxia and hemorrhagic shock on the interactions between plymorphonuclear neutrophils (PMNs) and cultured pulmonary artery endothelial cells (PAECs) were studied. The plasma samples were obt...The plasma effects during hypoxia and hemorrhagic shock on the interactions between plymorphonuclear neutrophils (PMNs) and cultured pulmonary artery endothelial cells (PAECs) were studied. The plasma samples were obtained from the goats under the following conditions: (1)Normal control plasma was obtained from the goats at sea level to aserve as the control (CP). (2)Hypoxic plasma was obtained after the goats were exposed to a simulated altitude of 4 000 m for 24 h (HP). (3) Hypotensive hypoxic plasma was obtained after the goats were bled to a mean arterial pressure of 5. 5±0. 3 kpa for 1 h under hypoxic condition (HHP). (4) Retransfused hypoxic plasma was obtained when the hypotensive goats were transfused with the shed blood for 4 h under hypoxic condition (RHP). It was found that HP , HHP and RHP especially RHP exerted profound effects on the activities of PMNs and PAECs in a concentration and time dependent manner after the PMNs and PAECs were incubated in the media containing different concentrations of the 4 kinds of plasma for different durations. Low concentration of RHP (less than 12. 5%) significantly increased the activity of PAECs (P<0. 01 vs CP) but its high concentration (more than 12. 5%) markedly decreased their activity (P<0. 01 vs CP, HP and HHP). HP, HHP and RHP increased the activity of PAECs in the early stage of incubation (1 to 3 h) (P<0. 01 vs CP) but decreased it in the late stage (6 to 12 h) (P<0. 01 vs CP). The activity of PMNs was significantly increased after 1 h incubation with HP, HHP and RHP (PM0. 001) and this effect was also concentrationdependent.The effects of RHP was the most potent, HHP the next and HP the least. The deformability of PMNs was significantly decreased (P <0. 001) after they were incubated in RHP for 3 h. The adhesive force of PMNs and PAECs was also significantly increased after 12 h incubation with RHP. These findings suggest that there are substances in the hypoxic plasma to activate or damage the interactions between PMNs and PAECs and the amounts of the substances are further increased in hypotensive hypoxic plasma and retransfused hypoxic plasma and the 'activation-damage'to PMNs and PAECs and the subsequent interactions between PMNs and PAECs play an important role in the pathological changes of hypoxia and hemorrhagic shock.展开更多
The monocellular adhesive ability of polymorphonuclear neutrophils (PMN)to vascular endothelial cells (VEC) was observed with micromanipulation technique when the cells were treated with endotoxin, Anti-CD18 monoclona...The monocellular adhesive ability of polymorphonuclear neutrophils (PMN)to vascular endothelial cells (VEC) was observed with micromanipulation technique when the cells were treated with endotoxin, Anti-CD18 monoclonal antibodies (McAb),anti-endothelial-leucocytic adhesion molecule-l (ELAM-l)McAb and anti-intercellular adhesion molecule-1 (ICAM-1) McAb and the effects of these 3 adhesion molecules on the development of adhesion force between PMNs and VECs were investigated. It was found that CD18 McAb decreased obviously the adhesive ability of PMNs in the whole course of adhesion increasing and the McAbs of ELAM-1 and ICAMI-1 inhibited most of adhesive force of VECs in the 4th and 12th hour of the course respectively.These findings suggest that the main molecular basis of rapid increase of adhesive ability of PMNs is CD18 expression and that of delayed increase of adhesive ability of VECs is ELAM-1 and ICAM-1 expression.展开更多
Objective: To investigate the effects of CD11/CD18 on the endothelial cell (EC)damages induced by polymorphonuclear neutrophils (PMN )from patients in the early stage postburn. Metbods: PMNs from 10 patients in the fi...Objective: To investigate the effects of CD11/CD18 on the endothelial cell (EC)damages induced by polymorphonuclear neutrophils (PMN )from patients in the early stage postburn. Metbods: PMNs from 10 patients in the first 24 to 48 h postburn were incubated with cultured human umbilical vein ECs (HUVECs) for 24 h and the percentage of PMN-EC adhesion was calculated, the content of lactate dehydrogenase (LDH), angiotensin converting enzyme (ACE) and 6-keto-PGF,. in the medium determined and the morphological changes Qf ECs observed. Results: After incubation with PMNs from burn victims, deformation of the HUVECs cell shrinkage, and formation of intercellurar space occurred and some cells were detached from the vascular wall. The content of LDH, ACE and 6-keto-PGF,. increased. When the PMNs were pretreated with monoclonal antibodies (McAbs)against CD11a/CD11b or CDl1b/CD18, the PMN-EC adhesion rate and the content of LDH, ACE and 6-ketoPGF,. in the culture medium decreased and Ec damages attenuated especially when the 2 kinds of McAbs were used in combination. Conclusion: The findings indicate that in the early stage postburn, PMNs can induce damages of ECs which results from CD1l/CD18 mediated PMN-CE adhesion and that the administration of McAbs against CD11/CD18 mediated PMN-EC adhesion and that the administration of McAbs against CD11/CD18 is able to b1ock partially the PMN-EC adhesion in burn patients and attenuate the damage of ECs.展开更多
OBJECTIVE: To study the ehanges of platelet endothelial cell adhesion molecule-1 (PECAM-1) expression on polymorphonuclear leukocytes (PMNs) in peripheral circulation anti pancreatic microcirculation in rats with acut...OBJECTIVE: To study the ehanges of platelet endothelial cell adhesion molecule-1 (PECAM-1) expression on polymorphonuclear leukocytes (PMNs) in peripheral circulation anti pancreatic microcirculation in rats with acute edematous pancreatitis (AEP). METHODS: The model of AEP was established with 50 Wistar rats, and the changes of PECAM-1 expression on PMNs from the splenic vein and inferior vena cava were determined by flow cytometry. RESULTS: PECAM-I expression on PMNs showed no significant difference between pancreatic microcirculation and peripheral circulation at AEP2h and AEP4h time points. From the AEP4h to the AEP8h time point, PECAM-1 expression in peripheral circulation was up-regulated, but PECAM-1 expression in pancreatic microcirculation was down-regulated. PECAM-1 expression had a significant difference between pancreatic microcirculation and peripheral circulation at the AEP8h time point (P<0.05). CONCLUSION: PECAM-1 expression on PMNs is in a converse way between pancreatic microcirculation and peripheral circulation in AEP.展开更多
AIM： To evaluate the accuracy of automated blood cell counters for ascitic polymorphonuclear （PMN） determination for： （1） diagnosis, （2） efficacy of the ongoing antibiotic therapy, and （3） resolution of spon...AIM： To evaluate the accuracy of automated blood cell counters for ascitic polymorphonuclear （PMN） determination for： （1） diagnosis, （2） efficacy of the ongoing antibiotic therapy, and （3） resolution of spontaneous bacterial peritonitis （SBP）. METHODS： One hundred and twelve ascitic fluid samples were collected from 52 consecutive cirrhotic patients, 16 of them with SBP. The agreement between the manual and the automated method for PMN count was assessed. The sensitivity/specificity and the positive/negative predictive value of the automated blood cell counter were also calculated by considering the manual method as the ＂gold standard＂ RESULTS： The mean ＋ SD of the difference between manual and automated measurements was 7.8 4- 58 cells/ram3, while the limits of agreement were ＋124 cells/mm3 [95% confidence interval （CI）： ＋145 to ＋103] and -108 cells/mm3 （95% CI： -87 to -129）. The automated cell counter had a sensitivity of 100% and a specificity of 97.7% in diagnosing SBP, and a sensitivity of 91% and a specificity of 100% for the efficacy of the ongoing antibiotic therapy. The two methods showed a complete agreement for the resolution of infection. CONCLUSION： Automated cell counters not only have a good diagnostic accuracy, but are also very effectivein monitoring the antibiotic treatment in patients with SBP. Because of their quicker performance, they should replace the manual counting for PMN determination in the ascitic fluid of patients with SBP.展开更多
Ceruloplasmin (CP) in peripheral leucocytes was determined by radioimmunoassay.It was observed that leucocytes of normal persons and leukemic patients contained CP. TheCP contents of polymorphonuclear cells (PMN) and ...Ceruloplasmin (CP) in peripheral leucocytes was determined by radioimmunoassay.It was observed that leucocytes of normal persons and leukemic patients contained CP. TheCP contents of polymorphonuclear cells (PMN) and blast cells of acute leukemia not in re-mission were both significantly higher than that of normal (P【0.05) and than that of acuteleukemia complete remission (P【0.05).展开更多
基金Supported by Unrestricted Research Grants (to Burri E) by the Freiwillige Akademische Gesellschaft (Basel, Switzerland) and the Gottfried und Julia Bangerter-Rhyner-Stiftung (Bern,Switzerland)Bühlmann Laboratories AG (Sch nenbuch, Switzerlanfd) provided the assays to measure ascitic calprotectin
文摘AIM: To evaluate the diagnostic capability of calprotectin in ascitic fluid for detecting a polymorphonuclear (PMN) cell count > 250/μL ascites. METHODS: In this prospective observational study, a total of 130 ascites samples were analysed from 71 consecutive patients referred for paracentesis. Total and differential leukocyte cell counts were determined manually with a Neubauer chamber and gentianviolet stain. Calprotectin was measured in 1 mL ascetic fluid by enzyme-linked immunosorbent assay (ELISA) and a point-of-care (POC) lateral flow assay with the Quantum Blue Reader (Bühlmann Laboratories). All measurements were carried out in a central laboratory by senior personnel blinded to patient history. A PMN count > 250/μL was the primary endpoint of the study. The diagnostic value of ascitic calprotectin measurement was assessed by comparing to the final diagnosis of each patient that had been adjudicated by investigators blinded to calprotectin values. RESULTS: The PMN count was > 250/μL in 19 samples (14.6%) from 15 patients (21.1%) and varied widely among the study population (range 10-19 800/mL and 1-17 820/mL, respectively). Spontaneous bacterial peritonitis (SBP) was the final diagnosis in four patients (5.6%). All patients with PMN ≤ 250/μL had negative bacterial culture. PMN count was elevated in five patients with peritoneal carcinomatosis, three with lymphoma, one with neuroendocrine carcinoma, and two with secondary peritonitis due to abdominal perforation. PMN cell counts correlated with ascitic calprotectin values (Spearman's rho; r = 0.457 for ELISA, r = 0.473 for POC). A considerable range of ascitic calprotectin concentrations was detected by ELISA [median 0.43 μg/mL, interquartile range (IQR) 0.23-1.23 (range 0.10-14.93)] and POC [median 0.38 μg/mL, IQR 0.38-0.56 (range 0.38-13.31)]. Ascitic calprotectin levels were higher in samples with PMN > 250/μL, by both ELISA [median (IQR) 2.48 μg/mL (1.61-3.65) vs 0.10 μg/mL (0.10-0.36), P < 0.001] and POC [2.78 μg/mL (2.05-5.37) vs 0.38 μg/mL (0.38-0.41), P < 0.001]. The area under the receiver operating characteristics curve for identifying an elevated PMN count was 0.977 (95%CI: 0.933 to 0.995) for ELISA and 0.982 (95%CI: 0.942 to 0.997) for POC (P = 0.246 vs ELISA). Using the optimal cut-off value for ELISA (0.63 μg/mL), ascitic calprotectin had 94.8% sensitivity, 89.2% specificity, positive and negative likelihood ratios of 8.76 and 0.06 respectively, positive and negative predictive values of 60.0% and 99.0% respectively, and 90.0% overall accuracy. Using the optimal cut-off value for POC (0.51 μg/mL), the respective values were 100.0%, 84.7%, 6.53, 0.00, 52.8%, 100% and 87.7%. Correlation between ELISA and POC was excellent (r = 0.873, P < 0.001). The mean ± SD of the difference was -0.11 ± 0.48 μg/mL with limits of agreement of + 0.8 μg/mL (95%CI: 0.69 to 0.98) and -1.1 μg/mL (95%CI: -1.19 to -0.91). CONCLUSION: Ascitic calprotectin reliably predicts PMN count > 250/μL, which may prove useful in the diagnosis of SBP, especially with a readily available bedside testing device.
文摘The plasma effects during hypoxia and hemorrhagic shock on the interactions between plymorphonuclear neutrophils (PMNs) and cultured pulmonary artery endothelial cells (PAECs) were studied. The plasma samples were obtained from the goats under the following conditions: (1)Normal control plasma was obtained from the goats at sea level to aserve as the control (CP). (2)Hypoxic plasma was obtained after the goats were exposed to a simulated altitude of 4 000 m for 24 h (HP). (3) Hypotensive hypoxic plasma was obtained after the goats were bled to a mean arterial pressure of 5. 5±0. 3 kpa for 1 h under hypoxic condition (HHP). (4) Retransfused hypoxic plasma was obtained when the hypotensive goats were transfused with the shed blood for 4 h under hypoxic condition (RHP). It was found that HP , HHP and RHP especially RHP exerted profound effects on the activities of PMNs and PAECs in a concentration and time dependent manner after the PMNs and PAECs were incubated in the media containing different concentrations of the 4 kinds of plasma for different durations. Low concentration of RHP (less than 12. 5%) significantly increased the activity of PAECs (P<0. 01 vs CP) but its high concentration (more than 12. 5%) markedly decreased their activity (P<0. 01 vs CP, HP and HHP). HP, HHP and RHP increased the activity of PAECs in the early stage of incubation (1 to 3 h) (P<0. 01 vs CP) but decreased it in the late stage (6 to 12 h) (P<0. 01 vs CP). The activity of PMNs was significantly increased after 1 h incubation with HP, HHP and RHP (PM0. 001) and this effect was also concentrationdependent.The effects of RHP was the most potent, HHP the next and HP the least. The deformability of PMNs was significantly decreased (P <0. 001) after they were incubated in RHP for 3 h. The adhesive force of PMNs and PAECs was also significantly increased after 12 h incubation with RHP. These findings suggest that there are substances in the hypoxic plasma to activate or damage the interactions between PMNs and PAECs and the amounts of the substances are further increased in hypotensive hypoxic plasma and retransfused hypoxic plasma and the 'activation-damage'to PMNs and PAECs and the subsequent interactions between PMNs and PAECs play an important role in the pathological changes of hypoxia and hemorrhagic shock.
文摘The monocellular adhesive ability of polymorphonuclear neutrophils (PMN)to vascular endothelial cells (VEC) was observed with micromanipulation technique when the cells were treated with endotoxin, Anti-CD18 monoclonal antibodies (McAb),anti-endothelial-leucocytic adhesion molecule-l (ELAM-l)McAb and anti-intercellular adhesion molecule-1 (ICAM-1) McAb and the effects of these 3 adhesion molecules on the development of adhesion force between PMNs and VECs were investigated. It was found that CD18 McAb decreased obviously the adhesive ability of PMNs in the whole course of adhesion increasing and the McAbs of ELAM-1 and ICAMI-1 inhibited most of adhesive force of VECs in the 4th and 12th hour of the course respectively.These findings suggest that the main molecular basis of rapid increase of adhesive ability of PMNs is CD18 expression and that of delayed increase of adhesive ability of VECs is ELAM-1 and ICAM-1 expression.
文摘Objective: To investigate the effects of CD11/CD18 on the endothelial cell (EC)damages induced by polymorphonuclear neutrophils (PMN )from patients in the early stage postburn. Metbods: PMNs from 10 patients in the first 24 to 48 h postburn were incubated with cultured human umbilical vein ECs (HUVECs) for 24 h and the percentage of PMN-EC adhesion was calculated, the content of lactate dehydrogenase (LDH), angiotensin converting enzyme (ACE) and 6-keto-PGF,. in the medium determined and the morphological changes Qf ECs observed. Results: After incubation with PMNs from burn victims, deformation of the HUVECs cell shrinkage, and formation of intercellurar space occurred and some cells were detached from the vascular wall. The content of LDH, ACE and 6-keto-PGF,. increased. When the PMNs were pretreated with monoclonal antibodies (McAbs)against CD11a/CD11b or CDl1b/CD18, the PMN-EC adhesion rate and the content of LDH, ACE and 6-ketoPGF,. in the culture medium decreased and Ec damages attenuated especially when the 2 kinds of McAbs were used in combination. Conclusion: The findings indicate that in the early stage postburn, PMNs can induce damages of ECs which results from CD1l/CD18 mediated PMN-CE adhesion and that the administration of McAbs against CD11/CD18 mediated PMN-EC adhesion and that the administration of McAbs against CD11/CD18 is able to b1ock partially the PMN-EC adhesion in burn patients and attenuate the damage of ECs.
文摘OBJECTIVE: To study the ehanges of platelet endothelial cell adhesion molecule-1 (PECAM-1) expression on polymorphonuclear leukocytes (PMNs) in peripheral circulation anti pancreatic microcirculation in rats with acute edematous pancreatitis (AEP). METHODS: The model of AEP was established with 50 Wistar rats, and the changes of PECAM-1 expression on PMNs from the splenic vein and inferior vena cava were determined by flow cytometry. RESULTS: PECAM-I expression on PMNs showed no significant difference between pancreatic microcirculation and peripheral circulation at AEP2h and AEP4h time points. From the AEP4h to the AEP8h time point, PECAM-1 expression in peripheral circulation was up-regulated, but PECAM-1 expression in pancreatic microcirculation was down-regulated. PECAM-1 expression had a significant difference between pancreatic microcirculation and peripheral circulation at the AEP8h time point (P<0.05). CONCLUSION: PECAM-1 expression on PMNs is in a converse way between pancreatic microcirculation and peripheral circulation in AEP.
文摘AIM： To evaluate the accuracy of automated blood cell counters for ascitic polymorphonuclear （PMN） determination for： （1） diagnosis, （2） efficacy of the ongoing antibiotic therapy, and （3） resolution of spontaneous bacterial peritonitis （SBP）. METHODS： One hundred and twelve ascitic fluid samples were collected from 52 consecutive cirrhotic patients, 16 of them with SBP. The agreement between the manual and the automated method for PMN count was assessed. The sensitivity/specificity and the positive/negative predictive value of the automated blood cell counter were also calculated by considering the manual method as the ＂gold standard＂ RESULTS： The mean ＋ SD of the difference between manual and automated measurements was 7.8 4- 58 cells/ram3, while the limits of agreement were ＋124 cells/mm3 [95% confidence interval （CI）： ＋145 to ＋103] and -108 cells/mm3 （95% CI： -87 to -129）. The automated cell counter had a sensitivity of 100% and a specificity of 97.7% in diagnosing SBP, and a sensitivity of 91% and a specificity of 100% for the efficacy of the ongoing antibiotic therapy. The two methods showed a complete agreement for the resolution of infection. CONCLUSION： Automated cell counters not only have a good diagnostic accuracy, but are also very effectivein monitoring the antibiotic treatment in patients with SBP. Because of their quicker performance, they should replace the manual counting for PMN determination in the ascitic fluid of patients with SBP.
文摘Ceruloplasmin (CP) in peripheral leucocytes was determined by radioimmunoassay.It was observed that leucocytes of normal persons and leukemic patients contained CP. TheCP contents of polymorphonuclear cells (PMN) and blast cells of acute leukemia not in re-mission were both significantly higher than that of normal (P【0.05) and than that of acuteleukemia complete remission (P【0.05).
