三种免疫学方法对氢化可的松抑制效应的评价

本文用溶血空斑试验玻片小室法和改良平皿法及溶血分光光度计测定法测定药用氢化可的松对小鼠 B 细胞功能的抑制效应。结果表明溶血分光光度计法最敏感、客观;溶血空斑试验改良平皿法较玻片小室法重复性好,标本可长期保存。是临床科研检测 B 细胞功能的可靠方法。     

Resistance towards calcium induced bilirubin dependent hemolysis in porcine erythrocytes

Hemolysis of erythrocytes from human and porcine blood was studied at different calcium chloride (CaCl₂) concentrations (0.04–1.83 mM) and at a constant bilirubin concentration (72 mM). Although, human erythrocytes showed significant hemolysis (64%) at the highest CaCl₂ concentration (1.83 mM) used in this study, remarkable resistance to this phenomenon was observed with porcine erythrocytes as only 11% hemolysis was observed. A similar pattern in the behavior of both human and porcine erythrocytes was observed when parameter such as bilirubin concentration or time of incubation with bilirubin was varied. Other divalent cations such as Mn²⁺, Ba²⁺ and Mg²⁺ were either least effective or ineffective in inducing hemolysis in presence of bilirubin. Serum albumin played a protective role in this phenomenon in a concentration dependent manner, as no hemolysis was observed at a bilirubin/albumin molar ratio of 1:1 or less. Differences in the structural make-up of proteins and lipids in the erythrocyte membranes of the two species may account for their different behavior.     

Influence of blood specimen collection method on various preanalytical sample quality indicators

Preanalytical errors contribute to a large proportion of total laboratory errors. In order to achieve continuous laboratory improvement, it is important to focus on all phases of patient specimen testing i.e. preanalytical, analytical and post-analytical. With large variations in the way venous blood specimens are collected using diverse devices in the country, the effect of such practices on specimen quality is not known. The purpose of this study was to monitor fourteen specimen preanalytical quality indicators in order to compare the usage of evacuated blood collection devices with needle and syringe open collection using either disposable tubes or re-washed glass vials. The study involved 26638 patient specimens assessed over a period of 6 months. The results demonstrated that evacuated closed blood collection resulted in an approximate 100-fold reduction in the incidence of hemolysis in samples. Similarly, there was a 200-fold reduction in incidence of insufficient specimen quantity while using evacuated collection system. It was also found that incidence of specimen contamination, improper volume of sample collected, and specimen spillage was also lower when the evacuated collection system was used. Further, it was also observed that the facility with a laboratory information system demonstrated much lower specimen identification and related errors. The observed results clearly demonstrate that the usage of the evacuated blood collection system resulted in improvement of preanalytical specimen quality as compared to needle and syringe usage.     

Effect of Oxidized Phosphatidylcholine on Biomarkers of Oxidative Stress in Rats

In this study it was planned to investigate the effect of oxidized phosphatidylcholine (derived from egg) feeding on lipid peroxidation of different tissues in rats. Male Wistar albino rats were fed oxidized and unoxidized phosphatidylcholine for 2 and 4 weeks, respectively. During the period of study food intake and body weights of animals increased gradually. Animals fed oxidized phosphatidylcholine for 2 and 4 weeks showed 33 and 15% spontaneous hemolysis of red blood cells in vitro. Under identical experimental conditions animals given unoxidized phosphatidylcholine showed 14.5 and 13.4% hemolysis for 2 and 4 week’s period, respectively. Thiobarbituric acid reactive substances (TBARS) level in thymus, spleen, kidney, heart, liver and lung significantly increased in rats given oxidized phosphatidylcholine as compared to unoxidized group. Furthermore, in oxidized phosphatidylcholine group TBARS values in kidney, liver and lungs continued to rise for 4 weeks of treatment while TBARS level in heart, spleen and thymus was found to be decreased at the end of 4 weeks of oxidized phosphatidylcholine feeding. Plasma triacylglycerol and cholesterol was found to increase in rats who had received oxidized phosphatidylcholine for 2 weeks. These findings suggest that excess and persistent intake of oxidized phosphatidylcholine can cause significant damage to organs.     

Extracellular Methemoglobin Mediated Early ROS Spike Triggers Osmotic Fragility and RBC Destruction: An Insight into the Enhanced Hemolysis During Malaria

Malaria infection is known to cause severe hemolysis due to production of abnormal RBCs and enhanced RBC destruction through apoptosis. Infected RBC lysis exposes uninfected RBC to the large amount of pro-oxidant molecules such as methemoglobin. Methemoglobin (MetHb) exposure dose dependently makes RBCs susceptible to osmotic stress and causes hemolysis. MetHb mediated oxidative stress in RBC correlated well with osmotic fragility and hemolysis. Interestingly, a reactive oxygen species (ROS) spike at 15 min was responsible for the observed effects on RBC cells. Two natural antioxidants N-acetyl cysteine and mannitol protected the RBC from MetHb-mediated defects, which clearly indicated involvement of oxidative stress in the process. MetHb due to its pseudo-peroxidase activity produces ROS in the external microenvironment. Therefore, classical peroxidase inhibitors were tested to probe peroxidase activity mediated ROS production with defects in RBCs. Clotrimazole (CLT), which irreversibly inactivates the MetHb (CLT-MetHb) and abolishes peroxidase activity, did not produce significant ROS outside RBC and was inefficient to cause osmotic fragility and hemolysis. Hence, initiating a chain reaction, MetHb released from ruptured RBC produces significant ROS in the external microenvironment to make RBC membrane leaky and enhanced hemolysis. Together data presented in the current work explored the role of MetHb in accelerated humorless during malaria which could be responsible for severe outcomes of pathological disorders.     

Interferences in clinical chemistry analysis

Analyses of body fluids in clinical chemistry laboratory are subject to a number of interferences that affect the analytical accuracy. The interferents arise from exogenous sources like drugs and additives as well as such endogenous sources like lipemia, hemolysis and icteria. Our studies demonstrate matrix interference in the form of analytical blas between serum and aqueous matrix calibrators. The clinical chemist should constantly be aware of this factor. Correction of interferences is recommended as an integral part of the quality assurance program.