低强度激光对人红细胞生物物理特性的影响

研究不同功率的低强度He-Ne激光对正常人体红细胞流变学特性影响。以正常人体红细胞为研究对象,测量了低强度He-Ne激光在不同照射时间、不同功率条件下红细胞的变形、取向、膜流动性、膜的微粘度和渗透脆性的变化情况。结果表明:照射后红细胞的变形性和膜流动性增强、渗透脆性下降。照射对红细胞流变学特性影响显著,其中激光能量为0.24 J、照射血样为2 mL时取得的照射效果最佳。     

在体网织红细胞微观流变学特性

用苯肼使动物造成急性溶血性贫血的方法,诱发动物体内新生网织红细胞大量增多,通过对新生网织红细胞的变形、取向及电泳率等指标的连续72h的测量,发现网织红细胞在转变为成熟红细胞的过程中,其流变学特性有明显改变.这对研究由于贫血等原因造成的网织红细胞增多情况下全血的微观流变学特性有重要的临床意义,同时也对新生网织红细胞的微观流变学特性进行了系统研究.     

CFU-E和其分化细胞的生物物理特性

用可诱发小鼠贫血的病毒(anemia-inducing strain friend’s virus,FVA)感染BALB／c小鼠,13d后取其脾脏,用Ficoll-Urografin分层液(1．070)分离出红系集落形成单位(colony forming unit-etythroid,CFU-E)细胞,用Wright-Giemsa染液染色并用透射电镜进行形态学观察,在培养液加细胞因子,如促红细胞生成素(erythropoietin,EPO)、白介素3(interleukin-3,IL-3)、干细胞刺激因子(stem cell factor,SCF),诱导其分化的情况下培养12、24和36h,分别对0、12、24和36h的细胞进行电泳率、膜的流动性和变形性的测量及红系特异转录因子GATA-1表达(0、12、24和48h)的检测,发现CFU-E细胞随着分化培养时间增加,其电泳率不断减少,膜的流动性不断增大,细胞的变形性和取向性逐渐变好;CFU-E在0、12和24hGATA持续高水平表达,而48h后,其表达明显降低．     

网织红细胞成熟过程中膜剪切弹性模量及表面粘度的改变

用鸡抗兔血清的抗体使兔造成急性溶血性贫血的方法,诱发兔体内同步生长的新生网织红细胞,用文宗曜等提出的一种测量红细胞膜剪切弹性模量及表面粘度的新方法——新型激光衍射法,连续72h监测经过不同发育阶段的网织红细胞的小变形指数和变形恢复过程(即松弛过程)中变形恢复到最大值一半的时间(即变形恢复半时间,t0.5),将测得的结果分别代入红细胞膜的剪切弹性模量公式和表面粘度公式。计算出不同发育阶段的网织红细胞的膜剪切弹性模量和表面粘度,发现网织红细胞在转变为成熟红细胞的过程中,其膜剪切弹性模量和表面粘度有明显改变。这对研究由于贫血等原因造成的网织红细胞增多情况下全血的微观流变学特性有重要的-临床意义,同时对新生网织红细胞在转化过程中膜的剪切弹性模量和表面粘度的变化规律加以系统研究,具有重要的基础理论研究价值。     

网织红细胞膜剪切弹性模量和表面粘度的研究

用苯肼使动物造成急性溶血性贫血的方法,诱发动物体内同步生长的新生网织红细胞,用一种测量红细胞膜剪切弹性模量及表面粘度的新方法——新型激光衍射法,连续72 h监测经过不同发育阶段的网织红细胞的小变形指数（DI）_d和变形恢复过程（即松弛过程）中变形恢复到最大值（DI）_max一半的时间t_0.5（即变形恢复半时间）,将测得的结果分别代入红细胞膜的剪切弹性模量（E）公式和表面粘度（μ_m）公式.计算出不同发育阶段的网织红细胞的膜剪切弹性模量和表面粘度,发现网织红细胞在转变为成熟红细胞的过程中,其膜剪切弹性模量和表面粘度有明显改变.这对研究由于贫血等原因造成的网织红细胞增多情况下,全血的微观流变学特性有重要的临床意义,同时对新生网织红细胞在转化过程中膜的剪切弹性模量和表面粘度的变化规律加以系统研究,填补了网织红细胞研究方面的空白,具有重要的基础理论研究价值.     

不同发育阶段红系造血细胞的生物力学特性和血液流变特性的变化

用可诱发小鼠贫血的病毒(anemia-inducing strain friend’s virus, FVA)感染BALB/c小鼠, 15 d后取其脾脏, 分离出原红细胞, 在加促红细胞生成素(EPO)等的培养基中培养12, 24, 48 h, 获得大量相对同步化的早幼红细胞、中幼红细胞、晚幼红细胞, 研究了不同发育阶段的上述早期红系造血细胞的生物力学及血液流变学特性的变化规律. 发现不同发育阶段的早期红系造血细胞, 随着发育其电泳率、渗透脆性、膜的流动性和黏弹性都发生了改变, 这种改变与膜脂的组成、膜蛋白、细胞膜的骨架蛋白、脂质分子与蛋白质分子的相互作用有关.     

蟾蜍红细胞与网织红细胞染色质蛋白的比较研究——Ⅰ、非组蛋白蛋白质

从亚洲蟾蜍(Bufo bufo asiaticus)的成熟红细胞与网织红细胞中分离纯化了染色质非组蛋白(NHP),并在体外转录系统中比较了两类细胞的NHP对RNA合成的激活作用。实验结果表明:从网织红细胞中分离获得的NHP只能部份恢复被网织红细胞组蛋白抑制的模板活力,而成熟红细胞的NHP不仅能恢复被成熟红细胞组蛋白抑制的模板活力,而且还能恢复被网织红细胞组蛋白抑制的模板活力。此外,从聚丙烯酰胺凝胶电泳图谱中也观察到这两类细胞的NHP的差异。     

60Co辐射对网织红细胞微观流变学特性的影响

采用60Co大剂量全身均匀急性辐射的方法,造成一种辐射贫血的动物模型.以便在几天内连续研究60Co辐射对新生的网织红细胞及红细胞流变学特性的影响.采用一种在低粘切变流场中能将红细胞变形指数DI分解为取向指数(DI)or和小变形指数(DI)d的新型激光衍射法,对网织红细胞及红细胞的变形指数、取向指数、综合变形指数(IDI)等血液流变学特性参数进行测量,发现在60Co大剂量辐射后,新生的网织红细胞及红细胞流变学特性存在明显异常.将这种60Co辐射造成的贫血模型与文宗曜等提出的用抗体诱导的大量同步化的球形红细胞贫血模型相比较,后者更具有明显的优点.同时为研究辐射对血液流变特性的影响及正确地挑选红细胞衰老模型提供了理论与实验的基础.     

严重高甘油三酯血症小鼠血液流变学的异常分析

目的:本研究旨在检测两种严重高甘油三酯血症(HTG)小鼠的血液流变学,建立具有异常血液流变学特点的HTG小鼠模型,以应用于进一步的相关机制研究。方法:采用ApoC3转基因与GPIHBP1基因敲除的HTG小鼠及野生型小鼠,所用小鼠均为C57BL/6J背景下8周龄雄性小鼠,通过生物物理法检测和比较其血浆粘度、红细胞渗透脆性、变形性与电泳率。结果:与野生小鼠相比,ApoC3转基因与GPIHBP1基因敲除的HTG小鼠血浆甘油三酯含量明显升高(P0.05),红细胞渗透脆性显著增加(P0.05),变形指数与电泳率明显降低(P0.05),血浆粘度显著升高(P0.05),但血细胞计数等血常规指标均未见显著差异(P0.05)。结论:Apo C3tg与GPIHBP1KO小鼠血液流变学发生异常改变,可能作为探讨HTG血液流变学异常和动脉粥样硬化心血管疾病病理机制的动物模型。     

运动中脂质过氧化物对人体红细胞膜结构和功能的影响

运动对红细胞某些特性的影响非常明显,其作用涉及到红细胞膜、代谢酶类和血红蛋白。先前的研究表明,运动中红细胞的破坏增多是引发运动性贫血的重要原因,但这些改变最终都归结于损伤红细胞膜的结构和功能,造成膜机械脆性和渗透性的增加,红细胞变形性降低,使其易于被破坏或被网状内皮系统所清除,从而导致运动性贫血的发生。 近来,愈来愈多的研究观察表明,氧自由基(OFR)引发的膜脂质过氧化反应是导致膜的液态性、流动性以及通透性改变,造成膜功能障碍。这为我们探索运动加速红细胞破坏、缩短红细胞寿命的机     

Biophysical and biorheological studies on the precursor cells at different stages

Splenic erythroblasts were obtained from mice during the acute disease caused by anemia-inducing virus (FVA). They were cultured in a medium containing EPO, BSA and so on. We studied their biomechanical and hemorheological behavior after 12 h, 24 h and 48 h. The results showed obvious changes in their electrophoretic mobility, osmotic fragility, membrane fluidity and membrane viscoelastic properties with the development of erythroblast. The changes were associated with the interactions of the protein and lipid and the elements of the membrane.     

Biophysical and biorheological studies on the precursor cells at different stages

Splenic erythroblasts were obtained from mice during the acute disease caused by anemia-inducing virus (FVA). They were cultured in a medium containing EPO, BSA and so on. We studied their biomechanical and hemorheological behavior after 12 h, 24 h and 48 h. The results showed obvious changes in their electrophoretic mobility, osmotic fragility, membrane fluidity and membrane viscoelastic properties with the development of erythroblast. The changes were associated with the interactions of the protein and lipid and the elements of the membrane.     

Comparison of membrane structure,osmotic fragility,and morphology of multiple sclerosis and normal erythrocytes

Erythrocyte membranes from multiple sclerosis (MS) patients and normal individuals were studied by electron spin resonance spectroscopy, osmotic fragility tests, scanning electron microscopy (SEM) and fatty acid analysis of membrane lipids. There was no significant difference in the membrane fluidity between MS and normal erythrocytes using fatty acid spin labels with the nitroxide moiety on carbons 5, 12, or 16 from the carboxyl group. Linoleic acid, which has been reported to decrease the absolute electrophoretic mobility of only MS erythrocytes, increased the fluidity of MS and normal erythrocyte membranes to a similar extent. The osmotic fragility of MS erythrocytes obtained from outpatients was similar to normal control cells but the osmotic fragility of erythrocytes obtained from hospitalized MS patients was greater than normal. Scanning electron microscopy of MS erythrocytes revealed no gross abnormalities. Cells incubated with linoleic acid had transformed from discocytes into sphero-echinocytes with prominent membrane surface indentations but MS and normal erythrocytes appeared identical. Of the fatty acid content of the total lipid extract, erythrocytes from most, but not all, MS hospitalized patients and some patients with other demyelinating diseases had relatively less (P<.001) 18:2 than the normal cells. These results indicate that at least some of the abnormalities reported in MS erythrocytes may only be found in hospitalized patients and may be due to other complications of the disease. They also indicate that the reported abnormal effects of linoleic acid on the electrophoretic mobility of MS erythrocytes may be caused by some other mechanism than an effect on the fluidity of the bilayer.     

Effects of Proteus mirabilis lipopolysaccharides with different O-polysaccharide structures on the plasma membrane of human erythrocytes

The effects of O33 and O49 P. mirabilis lipopolysaccharides (LPSs) on human erythrocyte membrane properties were examined. Physical parameters of the plasma membrane, such as membrane lipid fluidity, physical state of membrane proteins, and osmotic fragility, were determined. The fluidity of the lipids was estimated using three spin-labeled stearic acids of doxyl derivatives: 5-doxylstearic acid, 12-doxylstearic acid, and 16-doxylstearic acid. All the applied labels locate to different depths of the lipid layer and provide information on the ordering of phospholipid fatty acyl chain mobility. LPSs O49 increased the membrane lipid fluidity in the polar region of the lipid bilayer as indicated by spin-labeled 5-doxylstearic acid. An increase in fluidity was also observed in the deeper region using 12-doxylstearic acid only for O33 LPSs. The highest concentration of O33 LPSs (1 mg/ml) increased the motion of membrane proteins detected by the spin-label residue of iodoacetamide. These results showed different actions of O33 and O49 LPSs on the plasma membrane due to the different chemical structures of O-polysaccharides. P. mirabilis O33 and O49 LPSs did not induce changes in the membrane cytoskeleton, osmotic fragility and lipid peroxidation of erythrocytes. On the other hand a rise in the content of carbonyl compounds was observed for the highest concentrations of O33 LPS. This result indicated protein oxidation in the erythrocyte membrane. Lipid A, the hydrophobic part of LPS, did not change the membrane lipid fluidity and osmotic fragility of erythrocytes. Smooth and rough forms of P. mirabilis LPSs were tested for their abilities for complement-mediated immunohemolysis of erythrocytes. Only one out of seven LPSs used was a potent agent of complement-mediated hemolysis. It was rough, Ra-type of P. mirabilis R110 LPS. The O-polysaccharide-dependent scheme of reaction is presented.     

Alterations in human red blood cell membrane properties induced by the lipopolysaccharide from Proteus mirabilis S1959

The effect of lipopolysaccharide (LPS, endotoxin), isolated from Proteus mirabilis S1959 strain, on red blood cell (RBC) membranes in whole cells as well as on isolated membranes was studied. Lipid membrane fluidity, conformational state of membrane proteins and the osmotic fragility of RBCs were examined using electron paramagnetic resonance spectroscopy and spectrophotometric method. Lipid membrane fluidity was determined using three spin-labeled fatty acids: 5-, 12- and 16-doxylstearic acid (5-, 12- and 16-DS). The addition of LPS S1959 to RBC suspension resulted in an increase in membrane fluidity, as indicated by 12-DS. At the concentrations of 0.5 and 1 mg/ml, LPS treatment led to a significant (P<0.05) increase in lipid membrane fluidity in the deeper region of lipid bilayer (determined by 12-DS). The conformational changes in membrane proteins were determined using two covalently bound spin labels, 4-maleimido-2,2,6,6-tetramethylpiperidine-1-oxyl and 4-iodoacetamido-2,2,6,6-tetramethylpiperidine-1-oxyl (ISL). The highest concentration of endotoxin significantly (P<0.05) decreased the relative rotational correlation time of ISL and significantly (P<0.05) increased the osmotic fragility of RBCs. The effect of endotoxin was much more profound in isolated membranes than in intact cells treated with LPS. At the concentrations 0.5 and 1 mg/ml, LPS led to a significant increase in h(w)/h(s) ratio. These results indicated increased membrane protein mobility, mainly in the spectrin-actin complex in membrane cytoskeleton. These data suggest that LPS-induced alterations in membrane lipids and cytoskeleton proteins of RBCs lead to loss of membrane integrity.     

Microrheological characteristics of reticulocyte in vivo

Using the method of phenylhydrazine-induced anemia in rabbits, a mass of new reticulocytes, which synchronously grow, were got in vivo. The measurements of deformation index, orientation index, electrophoresis mobility etc. were performed for more than 72 h in the process of reticulocytes turning into red blood cells in vivo. There were obvious changes in the micro- rheological characteristics of reticulocytes in the course of turning into erythrocytes. The present study is significant in clinic for studying erythrocytes' microrheological characteristics when there are a lot of reticulocytes in blood, and also important in basic theorem for studying reticulocytes microrheological characteristics. It makes up a deficiency in the study on microrheological characteristics of reticulocytes turning into new RBCs from reticulocytes during reticulocytes life span.     

Microrheological characteristics of reticulocyte in vivo

Using the method of phenylhydrazine-induced anemia in rabbits, a mass of new reticu-locytes, which synchronously grow, were got in vivo. The measurements of deformation index, orientation index, electrophoresis mobility etc. were performed for more than 72 h in the process of reticulocytes turning into red blood cells in vivo. There were obvious changes in the micro-rheological characteristics of reticulocytes in the course of turning into erythrocytes. The present study is significant in clinic for studying erythrocytes' microrheological characteristics when there are a lot of reticulocytes in blood, and also important in basic theorem for studying reticulocytes microrheological characteristics. It makes up a deficiency in the study on microrheological characteristics of reticulocytes turning into new RBCs from reticulocytes during reticulocytes life span.     

Microrheological characteristics of reticulocyte in vivo

Using the method of phenylhydrazine-induced anemia in rabbits, a mass of new reticulocytes, which synchronously grow, were got in vivo. The measurements of deformation index, orientation index, electrophoresis mobility etc. were performed for more than 72 h in the process of reticulocytes turning into red blood cells in vivo. There were obvious changes in the microrheological characteristics of reticulocytes in the course of turning into erythrocytes. The present study is significant in clinic for studying erythrocytes’ microrheological characteristics when there are a lot of reticulocytes in blood, and also important in basic theorem for studying reticulocytes microrheological characteristics. It makes up a deficiency in the study on microrheological characteristics of reticulocytes turning into new RBCs from reticulocytes during reticulocytes life span.     

Estimation of cell membrane properties and erythrocyte red-ox balance in patients with metabolic syndrome

Metabolic syndrome (MS) is associated with occurrence of the many cardiovascular risk factors such as atherogenic dyslipidemia, visceral fat distribution, arterial hypertension and pro-thrombotic and pro-inflammatory status. In our study the effect of disorders that appear in MS on red-ox balance and erythrocyte cell membrane properties were estimated. The study comprised 50 patients with diagnosed MS and in 25 healthy subjects. Content of thiobarbituric acid reactive substances (TBARS) and catalase, superoxide dismutase and glutathione peroxidase activity were estimated in red blood cells. Moreover, conformation status of membrane proteins, membrane fluidity and osmotic fragility were evaluated. MS was found to manifest: (1) the increase of the concentration of TBARS in erythrocytes with no statistically significant differences in antioxidant enzymes activity, (2) disorders in the structure of erythrocyte cytoskeleton proteins, (3) the increase in membrane lipids fluidity at the depth of 5th and 12th carbon atom of fatty acid hydrocarbon chain and significantly decreased fluidity at the depth of 16th carbon atom, (4) increased erythrocyte osmotic fragility.