The Molecular Mechanism in the Damage of Erythrocyte Membrane of the Cows with Hemoglobinuria

The molecular mechanism in the damage of erythrocyte membrane of the cows with hemoglobinuria was studied with the field cases and the group comparison.The field cases were devided into three groups:the hemoglobinuria group(HG),the low phosphorous group(LPG)and the control group(CG).The content of phospholipid constituents in the erythrocyte membrane and the protein constituents in membrane skeleton were determined molecularly and the shape of erythrocyte was examined with the scanning electron microscope.The result showed that:(1)the concentration of phosphatidylethanolamine(PE)in HG was lower significantly than that in LPG and CG;the concentration of sphingomyeline(SM) and phosphatidylcholine(PC)+phosphatidylserine(PS) in HG was significantly lower than that in the other groups;the content of PC+PS was lower and the concentration of SM was higher in LPG with comparing that in CG,the significant positive correlation was observed between the concentration of phosphorus in serum and the concentration of PE,respectively.The significant negative correlation was observed between the content of phosphorus in serum and the content of SM,respectively.(2)the difference of protein constituent in membrane skeleton between LPG and CG was not found,however,the concentration of spectin in band Ⅰ,Ⅱ and the content of protein in band Ⅳ 2 was lower and the concentration of protein in band Ⅲ significantly higher in HG than that in LPG and CG(P<0.01);(3)the form of erythrocyte observed with scanning electron microscope changed from discal to the spinal,to spheric form,to hemolysis ultimately following the reduction of the concentration of phosphorus in serum.It was concluded that the change of the constituent of phospholipid in erythrocyte membrane and protein in membrane skeleton and the form of erythrocyte is the most important factors in hemolysis for hypophosphatemia.

Altered Erythrocyte Membrane Calcium Binding in Hypertensive Rats and the Effects of Sodium Tanshinone Ⅱ-A Sulphonate on It

The calcium binding of erythrocyte membrane was determined in spontaneous hypertensiverats (SHR)and renovascular hypertensive rats (RVHR two-kidney, one-clip model) and the effect ofsodium tanshinone Ⅱ-A sulfonate(DS-201)on the calcium binding in SHRs was investigated. Ourresults show that the basal calcium binding was reduced in SHRs (P<0.01 vs WKY),while the maximalcalcium binding was not,but both typies calcium bindings had no significant change in RVHRs.Sodiumtanshinone Ⅱ-A sulfonate (125μ mol/L)have no effect on the calcium binding of ecythrocyte membraneof SHR in vitro.These data further support the hypothesis that there is a cell membrane abnormalitypresent in SHRs which may possibly serve as a marker genetics of in hypertension.

The Relationships between Erythrocyte Membrane n-6 to n-3 Polyunsaturated Fatty Acids Ratio and Blood Lipids and C-reactive Protein in Chinese Adults:An Observational Study

Objective To investigate the relationships between erythrocyte membrane n-6：n-3 PUFAs ratio and blood lipids and high sensitivity C-reactive protein （hs-CRP）. Methods The Chinese and a observational study consisted of a population-based cross-sectiona subsequent 1-year follow-up study of 171 subjects with the fasting cholesterol of 5.13-8.00 mmol/L. study of 456 plasma total Results In the cross-sectional analysis, plasma low-density lipoprotein cholesterol （LDL-c） had a significant and negative association with the erythrocyte membrane n-6：n-3 PUFAs ratio （P for trend=0.019） after adjusting for sex, age and total PUFA percentage. In the follow-up study, 171 subjects were categorized into quartiles by the changes of n-6：n-3 ratio in erythrocyte membrane （△=month 12-month 0）. In the top quartile whose ratios of n-6：n-3 increased by an average of 1.25 during the follow-up, the LDL-c-lowering extent was 3.3 times of that in the lowest quartile whose ratios of n-6：n-3 decreased by an average of 1.13 （-1.07 mmol/L v.s. -0.32 mmol/L）. The hsCRP decreased by 0.11 mg/dL in the lowest quartile while increasing by 0.10 mg/dL in the top quartile （P for difference=0.052）. Conclusion Our results suggested that the balance between n-6 and n-3 fatty acids may optimize the cardiovascular benefits from dietary PUFAs.

Effect of Huoxuequyu Recipe on Erythrocyte Ultrastructure and Membrane ATPase Activity in Rats with Passive Smoking

The effect of 'Huoxuequyu' recipe on erythrocyte membrane Ca2+ATPase activity, Na+-K+-ATPase activity and erythrocyte ultrastructure in rats exposed to passive smoking were observed. The erythrocyte membrane Ca2+-ATPase activity and Na+-K+-ATPase activity were found to be decreased in the model group as compared with the control group (P<0.01, P<0.01), while no significant difference existed between the treated group and the control group (P>0.05). Our results also showed that there were more deformed red blood cells in model group than in control group under TCM and SEM (P<0.0001). No significant difference was found between control group and treated group (P>0.05). Furthermore,these findings indicated that 'Huoxuequyu' recipe , a prepared natural herbal medicine, may be involved in modulation of the activity of erythrocyte membrane enzymes,thereby protecting erythrocyte membrane, so as to improve the erythrocyte function.

Studies on the Interaction between Yttrium and Human Erythrocyte Membrane

Yttrium ions promote hemolysis of human erythrocytes, and it's critical hemolytic concentration is 3x10(-5) mol/L. The interactions between Y3+ and human erythrocyte membrane have been studied by SDS-PAGE, CD spectra and fluorescence techniques. The experimental results indicated that the high concentration of Y3+ altered the conformation of human erythrocyte membrane proteins and lead to cross-linking or polymerization among membrane proteins. Y3+ can quench the intrinsic fluorescence of membrane proteins. There are two kinds of binding site's of Y3+ to erythrocyte membrane, the binding capacities (n) and appearance binding constants (K) were determined to be: n(1) = 2.11 x 10(-3) mol/g membrane protein, n(2) = 3.56 x 10(-1) mol/g membrane protein; K-1 = 4.51 x 10(6) L/mol, K-2 = 1.18 x 10(6) L/mol. These results show that Y3+ ions interact strongly with human erythrocyte membrane, and appear obviously cell toxicity.

Changes in erythrocyte membrane ATPases and plasma lipid peroxides in upper abdominal surgery under intravenous procaine-balanced anesthesia

AIM To observe the changes in erythrocyte membrane ATPases and plasma lipid peroxides (LPO) patients with in abdominal surgery under intravenous procaine-balanced anesthesia.METHODS By determining the ATPase activities of erythrocyte membrane, effects of upper abdominal surgery under intravenous procaine-balanced anesthesia on the function of erythrocytes were observed in 15 patients undergoing cholecystectomy and gastrectomy (5 males and 10 females, aged 45.9±10.20 years and weighed 60.60kg±11.93kg). All patients were free from severe renal, hepatic, pulmonary, cardiac, metabolic and endocrinological diseases and acute infection for at least 2 weeks before surgery. Patients receiving any drug known to affect carbohydrate metabolism prior to anesthesia were excluded from the study.RESULTS Erythrocyte membrane Na+, K+-ATPase, Mg2+-ATPase, Ca2+, Mg2+-ATPase activities were not significantly changed 60min-90min after incision as compared with 30min before anesthesia, but were decreased markedly 10min and 24 hours after completion of operation (P＜0.01). Plasma lipid peroxides (LPO) were increased significantly 24 hours after surgery (P＜0.01) following an initially marked but transient reduction. Plasma LPO changes were not correlated with erythrocyte membrane ATPase activities, r=-0.0396, -0.0097 and 0.4383, respectively (P＞0.05).CONCLUSION Abdominal surgical trauma under intravenous procaine-balanced anesthesia may be associated with the decreased ATPase activities of erythrocyte membrane and increased LPO in plasma.

The investigation for the relationship among serum leptin, erythrocyte membrane Ca^(2+)-ATPase activity and hypertensive disorder complicating pregnancy

Objective： To study the significance of Leptin and the activity of erythrocyte membrane Ca^2＋-ATPase（EMCA） in the development of hypertensive disorder complicating pregnancy. Methods： Radioimmunoassay was used to test the level of serum Leptin, and the activity of EMCA was determined chemically in 38 pregnant women with hypertensive disorder complicating pregnancy and 36 normotensive pregnant women. Results： The level of serum Leptin in hypertensive disorder complicating pregnancy（gestational hypertension： 13.76 ± 3.46 ng/ml; preeclampsia： 15.76 ± 5.47 ng/ml; eclampsia： 18.32 ± 6.38 ng/ml）was significantly higher than that in normotensive pregnant women （11.33 ± 2.93 ng/ml） ,respectively. The average EMCA activity of patients with hypertensive disorder complicating pregnancy （gestational hypertension： 1.65 ± 0.24 μmol·pi/mg.h ; preeclampsia： 1.37 ± 0.19 μ mol·pi/mg·h;eclampsia：1.12 ± 0.14 μmol·pi/mg·h） was significantly lower than that of normotensive pregnant women（1.83 ± 0.38 μ mol·pi/mg·h）,respectively. There was a negative correlation between the level of serum Leptin and the activity of RMCA in hypertensive disorder complicating pregnancy （r = -0.63）. Conclusion： Inhibition of EMCA activity of erythrocyte in hypertensive disorder complicating pregnancy may increase cytoplasmic free calcium, which contributes to the development of hypertensive disorder complicating pregnancy. The negative correlation between the level of serum Leptin and the activity of EMCA, also suggested that serum Leptin and the activity of EMCA may play a role in the development of hypertensive disorder complicating pregnancy.

Interaction of Lanthanum and Calcium with Human Erythrocyte Membranes

The effect of lanthanum and calcium on the structure and function of human erythrocyte membranes was investigated by fluorescence polarization, spin- labeled electron spin resonance (ESR) and laser Raman spectroscopy. The results showed that low concentration of La3+ (0.5 mu mol/L) activated a Little (Na++K+)-ATPase and Mg2+-ATPase activities, and it inhibited obvi ously the ATPase activities with increasing its concentrations. La3+ lowered the lipid fluidity of human erythrocyte membranes and decreased the vibration intensity of alpha-helix of the protein in the Amide I '. The effect of Ca2+ on the lipid fluidity and alpha-helix of the protein in the Amide I ' was smaller than that of La3+.

Effect of "Jia Wei Fo Shou San" on Erythrocyte Membrane Calcium,Zinc Concentration in Pregnant Rats with Asymmetrical Intrauterine Growth Retardation Induced by Passive Smoking

Using experimental model of pregnant rats with asymmetrical intrauterine fetal growth retardation induced by passive smoking, the effects of natural herb 'Jia Wei Fo Shou San' on erythrocyte membrane calcium, zinc concentration were observed by atomic absorption spectrophotometry. The results showed that the mean fetal birth weight, zinc concentration of erythrocyte membrane were found to be decreased in the model group as compared with the control group (P＜0. 01, P＜0. 01). On the other hand, the element of calcium onto erythrocyte membrane were higher in model group than that in control group(P＜0. 05). These changes were significantly mild in the treated group and were similar to those of the control group (P＜0.05). Furthermore,our findings indicated that the zinc concentration of erythrocyte membrane seems to be positively correlated with the birth weight(P＜0. 01). Calcium composition of red cell membrane showed a significant negative relation to the birth weight (P＜0.05). Our results provided an experimental evidence that normal concentrations of calcium and zinc onto erythrocyte play an important part in fetal growth. One of the mechanisms of 'Jia Wei Fo Shou San' in improving fetal growth may have something to do with modulation of erythrocyte calcium,zinc element,thereby protecting bio-functions of erythrocyte and promoting blood circulation.

Changes in erythrocyte membrane ATPase and plasma lipid peroxides during perioperative period

Changes in erythrocyte membrane ATPases and plasma lipid peroxides（LPO） were studiedperioperatively in 15 abdominal surgical patients under intravenous procaine-balanced anesthesia.The resultsrevealed that erythrocyte membrne Na+-K+ATPase,Mg2+ATPase and Ca2+-Mg2+ATPase activities werenot significantly changed at 60～90 min after incision as compared with 30 min before anesthesia,but weredecreased markedly at 10 rain and 24 hours after completion of operation（P【0.01）.Plasma LPO wereincreased significantly by 24 hours after surgery（P【0.01） following an initially marked but transientreduction.Plasma LPO changes were not correlated with erythrocyte membrane ATPase activities,r=-0.0396,-0.0097 and-0.4383 respeetively（P】0.05）.These correlations are suggested to be inducedby neuroendoerine responses after anesthesia and/or surgical trauma.

Study on Phospholipid Composition of Erythrocyte Membrane in Hypophosphatemic Cows

The phospholipid constituents of the erythrocyte membrane of cows in hypophosophorus were detected with the field cases and the group comparison. The cows were divided into three groups: the hemoglobinuria group (HN), the hypophosphatemia group (HP) and the control group (CK). The content of the phospholipid constituent in HN and HP obviously changed: phosphatidylethaanolamine (PE) content in HN was significantly lower than that in HP and CK; but sphingomyline (SM) and phosphatidycholine (PC) + phosphatidylserine (PS) content in HN were significantly higher than that in the two other groups; in comparison between HP and CK, PC + PS content was lower and SM content was higher in HP; significant positive correlation and negative correlation were observed between serum phosphorus and PE content, serum phosphorus and SM content respectively.

Effect of Huoxuequyu Recipe on Erythrocyte Membrane Ca^(2+)-ATPase Activity in Pregnant Rats with Asymmetrical Intrauterine Growth Retardation Induced by Passive Smoking

The effect of Huoxuequyu recipe on erythrocyte membrane Ca2+-ATPase activity in pregnant rats with passive smoking induced asymmetrical intrauterine growth retardation(IUGR)was observed.The rats were divided into three groups:control group,model group and treated group.The fetal mean birth weight and erythrocyte membrane Ca2+-ATPase activity were found to be decreased in the model group as compared with the control and treated groups.There was a significant difference between the model group and the control group(P<0.01),while no significant difference existed between the treated group and the control group(P>0.05).Furthermore,the findings indicated that the erythrocyte membrane Ca2+-ATPase activity seem to be positively correlated with the birth weight(P<0.05).On the basis of our observation,it is believed that the Huoxuequyu decoction could increase erythrocyte membrane Ca2+-ATPase activity and promote blood circulation, thereby exerting a beneficial effect on fetal development.

Assessment of Membrane Erythrocyte Cholesterol Level in Sickle Cell Disease

Objective: The aim of this study was to assess the level of erythrocyte membrane cholesterol in sickle cell patients, which is one of the essential parameters of membrane fluidity that contributes to understanding the hemolytic state of the erythrocyte. Methods: We worked with blood specimens from 20 controls and 50 sickle cell patients. The blood count and the isoelectric focusing (IEF) were performed on the samples in order to select them. The titration of the erythrocyte membrane cholesterol was made after washing and lysing the erythrocytes with the hemolyzing solution (EDTA, 2 Mercapto-Ethanol, NADP, NaOH). The cholesterol level was assessed by the enzymatic colorimetric method. The results were analyzed by Student’s test. Results: We worked with 16 control subjects with a hemoglobin status and a normal hemogram. The evaluation of the erythrocyte membrane cholesterol level of the samples allows us to define the reference interval (α = 0.05) at 17.55 ± 3.83 mg of cholesterol/g of hemoglobin. The erythrocyte membrane cholesterol levels of sickle cell patients found in this study were 11.58 ± 2.98 mg cholesterol/g of hemoglobin. In this study, 38 sickle cell patients (76%) were found with a low erythrocyte membrane cholesterol level compared to the reference interval. Statistical analysis showed that there was a significant difference (α = 5%) between the erythrocyte membrane cholesterol level of normal subjects and sickle cell patients. Conclusion: Most of the sickle cells patients had a decreased erythrocyte membrane cholesterol level. This reduces the membrane fluidity making the erythrocyte membrane rigid and more fragile. This may be one of the factors responsible for the lyses of erythrocytes.

The Fusability of Erythrocytes as a Method for Evaluating the Instability of Their Membranes

A novel approach to the study of erythrocytes instability based on assessment fusability of their plasma membrane is presented. The fusability of erythrocytes normal blood, stored for 7 days at 4°C, and type 2 patients with diabetes were studied. Human erythrocytes were fused by incubation with La3+ at final concentrations from 50 to 200 μM. Erythrocytes aggregates were incubated at 37°C for 120 min. The fusability of erythrocytes was evaluated using a light microscope. It was shown that La3+ induced extensive fusion of erythrocytes after 7 days of blood storage and erythrocytes of patients with diabetes. Incubation of normal erythrocytes at 37°C for 120 min does not induce cell fusion.

SUPEROXIDE DISMUTASE ACTIVITY AND MEMBRANE LIPIDS COMPOSITION OF ERYTHROCYTES IN PATIENTS WITH DILATEDCARDIOMYOPATHY

Objective To rurther conflrm the role or lipld-peroxldation caused by oxygen free radicals injury played in the pathogenesis of dllsted cardlomypathy. MethOds The superoxide dismutase activities and lipids composltion of erythrocytes in 18 patients with dilated cardiomyopathy and 16 healthy controls were measured. Results Superoxide dlsmutase(SOD) activites of erythrocytes were lower in dilated cardiomyopathy(DCM) patients than that in healthy controls (P <o. oo1 ). The lipids composition of erythrocytes has changed in the DCM patients compared with healthy controls: total liplds changed little (P >o. o5); total phospholipids were lower, but not significantly (P>o. o5): total cholesterol increased significantly (P <o. o5). The cholesterol to phospholipids molecular ratio of erythrocyte membrane has increased remarkably (P >o. o5). Conclusion It can be supposed that decreased SOD activitles play an important role in the damage or membrane system and the pathogensis of DCM.

Erythrocyte Membrane Protein Band 3 Predicts Interferon Ribavirin-Induced Anemia

Aim & Background: It was proposed that the differences in erythrocyte membrane protein contents—especially band 4—take a role in the serious anemia related to interferon plus ribavirin (I/R). The aim of this study is to evaluate whether the erythrocyte membrane protein contents predict anemia related to I/R or not. Methods: 180 mcg interferon α 2a once a week and weight adjusted ribavirin daily were given for 48 weeks to fifty patients with chronic hepatitis C. It was diagnosed as anemia when haemoglobin concentration was Results: Anemia developed in 17 patients (34%). The levels of erythrocyte membrane proteins were as;spectrin: 20.468 ± 2.5902, ankyrin: 4.576 ± 1.2706, B3: 19.240 ± 2.8358, B4.1: 5.628 ± 1.8832, and B4.2: 5.848 ± 1.8030. When the relation between the development of anemia and erythrocyte membrane proteins was investigated, a relation was only found at B3 which was not statistically significant (p = 0.058). When ROC analysis was performed, 95% CI p = 0.035 for B3 (0.517 - 0.792) was found. In patients whose B3 level was below 17.7%, the sensitivity of anemia development risk was calculated as 64.7% and the specificity thereof was calculated as 66.7%. Erythrocyte membrane protein contents by gender were only different at B3 (p = 0.042). Anemia developed in 17 patients (34%). 14 of these patients were of female and 3 were of male gender;the gender played a significant role in terms of anemia (p = 0.003). Conclusions: Erythrocyte membrane protein B3 is not only useful in predicting the patient under the risk of developing anemia, but also it may be useful in preventing it and it may explain why women inclined to anemia.

Protective effect of Apocynum venetum (罗布麻) against lipid peroxidation damage of erythrocyte membrane

Objective: To investigate the protective effect of Apocynum venetum on lipid peroxidation damage of erythrocyte membrane. Methods: Model of lipid peroxidation of erythrocyte membrane was manufactured by three kinds of radicals generation systems. To set up a normal control group (NC),a model control group (MC),and four Apocynum venetum groups(A.V). Observation was made on the content of malondialdehyde (MDA). Results: Compared with MC group,all of the four Apocynum venetum groups dose-dependently inhibited the increases of MDA content in the membrane induced by xanthine - xanthine oxidase system,H 2O 2 or UV light. Conclusions: Apocynum venetum may protect erythrocyte membrane from the lipid peroxidative damage induced by radicals.

Relationship between the Contents of MDA, the Activity of SOD in Serum of the Milk Goat with Fluorosis and the Erythrocytic Membrane Liquidity

To research the relation among the erythrocytic membrane.liquidity and the contents of MDA, the activity of SOD in serum of the milk goat with fluorosis. An animal model with fluorosis was constructed, and the fluorescent probe technique of DPH was used to measue the erythrocytic membrane liquidity. At the same time, the contents of MDA and the activity of SOD in serum were measured. The results showed that the erythrocytic membrane liquidity in the control group and flurosis group were 5.6742 ± 0.4417 and 3.7248 ± 0.4521 （P 〈0.01） respectively, the contents of MDA in serum were 2.0408 ± 0.198 and 4.494± 0.438 （P 〈0.01） respectively, the activities of SOD were 175.638 ± 22.201 and 113.714 ± 34.258 （P 〈0.01） respectively. The correlation analysis indicated that the relation between the activity of SOD and the liquidity of erythrocytic membrane was positive correlation （ r= 0.7321, P 〈0.05）, whereas the relation between the contents of MDA and the liquidity of erythrocytic membrane was negative correlation （r = -0.6438, P 〈0.01）. The erythrocytic membrane liquidity decreased in milk goat with fluorosis, which played a role in the occurrence and development of the fluorosis. There was correlation among the erythrocytic membrane liquidity and the contents of MDA, the activity of SOD.

Erythrocyte membrane encapsulated gambogic acid nanoparticles as a therapeutic for hepatocellular carcinoma

Gambogic acid(GA)is a potential clinical anticancer drug that can exert antitumor effects via various molecular mechanisms.Notwithstanding,GA’s low water solubility,poor stability,short half-life,and unavoidable toxic side effects have significantly hampered its clinical application.Erythrocyte membranecoated nanoparticles(RBCM-NPs)improve drug’s physicochemical properties,biocompatibility,and pharmacokinetic behaviors,allowing for long-term drug circulation and passive targeting.In this study,a novel biomimetic drug delivery system(DDS)against hepatocellular carcinoma was prepared by covering RBCM on GPP-NPs(GA-loaded m PEG-PLA NPs)to develop the RBC@GPP-NPs.In comparison to RBCM-free nanoparticles and free GA,RBC@GPP-NPs improved the drug’s water solubility,stability,safety,and antitumor activity in vivo.We expect that this bionic nanoparticle composite can expand the clinical applicability of GA and provide a feasible solution for the research and development of GA’s nano-formulation.

Effects of L.F04,the Active Fraction of Lycopus lucidus, on Erythrocytes Rheological Property

Objective: To study the effects of L.F04, the active fraction of Lycopus lucidus, on erythrocytes rheological property so as to investigate its mechanism in promoting blood circulation and removing blood stasis. Method: The effects of L.F04 (used for treatment for 10 days in different dosages) on deformability, aggregation and membrane liquidity of erythrocytes (MLE) as well as whole blood apparent viscosity (η_b) were examined on the basis of rat model of blood-stasis syndrome induced by venous injection of high molecular weight dextran. Result: As compared with the normal control group, the model group's RBC deformability and MLE were lower, and the aggregation of erythrocytes and η_b were higher. Compared with the model group, both L.F04 0.612g/kg and 0.306g/kg showed significant effect in improving deformability and inhibiting aggregation of red blood cells (RBC) and reducing blood viscosity. The trend of improving MLE was also shown. Conclusion: L.F04 could significantly improve the abnormal rheological property of erythrocytes.