The efflux of nitro oxide (NO) in the duration of storing red blood cells (RBCs) was the main reason resulting in decrease and even loss of vasodilatory activity, cell deformability and ability of carrying oxygen (O2) in the stored RBCs. The deep understanding physical functions and acting ways of NO in circulatory system, as well as transformations and balance control of S-Nitrosohemoglobin (SNO-Hb) has an important significance for ensuring sure safety and efficacy of transfusion. In this article, the physical functions, acting ways, retaining and transferring form of nitro oxide, and SNO-Hb adjusting, as well as effects of SNO-Hb concentration on change on stored red blood cells were reviewed.
Erythrocytes ; metabolism ; physiology ; Hemoglobins ; biosynthesis ; Humans ; Nitric Oxide ; metabolism

OBJECTIVE: To investigate the release of exosome (Exo) from leukocyte-depleted red cell suspension (LDRCS) at different storage time and its regulation on proliferation of hematological tumor cells and possible mechanism. METHODS: The Exo (RBC-Exo) in LDRCS at different storage time was obtained by ultracentrifugation, and the morphology and immunological marker of RBC-Exo were detected by transmission electron microscopy and Western blot, respectively. The particle size distribution of RBC-Exo in LDRCS at different storage time was detected by Dynamic Light Scattering. CCK-8 assay was used to explore the effect of RBC-Exo on hematological tumor cell proliferation. Western blot was used to detect the expression of proliferation-related proteins in hematological tumor cells after co-culture with RBC-Exo. RESULTS: RBC-Exo was isolated, which was characterized by cup-like shape, particle size distribution ranged from 20 to 200 nm, CD63/TSG101 enriched, Calnexin negative, CD235a positive and CD41 negative. The particle size distribution of RBC-Exo from LDRCS between middle was not significantly different and late stored stage. But the particle size distribution of RBC-Exo at middle-late stored stage(>14 d) was larger than that at early stored stage (≤14 days). Compared with the control group, RBC-Exo could significantly promote the proliferation of HBL1, U2932 and Jurkat cells. Compared with the control group, the cycle-related protein P21 was significantly down-regulated in HBL1, U2932 and Jurkat cells after co-culture with RBC-Exo for 3 days, while the anti-apoptotic protein BCL-2 was not changed significantly. CONCLUSION The morphology of RBC-Exo from LDRCS at middle-late stored stage was different from that at early stored stage. RBC-Exo could promote the proliferation of hematological tumor cells, possibly by regulating the expression of cycle-associated protein P21.
Cell Proliferation ; Erythrocytes ; Exosomes/metabolism* ; Hematologic Neoplasms/metabolism* ; Humans ; Leukocytes

The interaction of calcium and local anesthetics was investigated with the lipid extracted human RBC membrane fragments treated with carbodiimide in order to titrate carboxyl groups. A water soluble carbodiimide [1-cyclohexyl-3-(2-morpholinoethyl) carbodiimide methotoluene-p-sulfonate], referred to as a carbodiimide reagent, and glycine methylester were used for this purpose. About 76% of carboxyl groups of the fragments were modified at a concentration of 0.05M carbodiimide reagent. The interaction of calcium and local anesthetics such as procaine and lidocaine with these fragments still showed typical competition. However, when the calcium binding was decreased to 8% at a higher concentration of carbodiimide reagent (0.08M), the local anesthetics still inhibited the calcium binding, but were not competitive in nature. In other words, if concentrations of the carbodiimide reagent were raised, the degree of inhibition by the local anesthetics was gradually decreased and was not competitive in nature. Finally, no inhibition was demonstrated when the concentration of the reagent was 0.1 to 0.4M. The above findings, seem to suggest that local anesthetics such as procaine and lidocaine interact with carboxyl groups, in addition to phosphodiester groups of phospholipids as previously reported, and inhibited competitively calcium binding to carboxyl groups of the membrane fragments.
Anesthetics, Local/pharmacology* ; Calcium/metabolism* ; Carbodiimides/pharmacology* ; Cell Membrane/metabolism ; Erythrocytes/metabolism* ; Human ; In Vitro ; Protein Binding

The interaction of calcium and local anesthetics was investigated with the lipid extracted human RBC membrane fragments treated with carbodiimide in order to titrate carboxyl groups. A water soluble carbodiimide [1-cyclohexyl-3-(2-morpholinoethyl) carbodiimide methotoluene-p-sulfonate], referred to as a carbodiimide reagent, and glycine methylester were used for this purpose. About 76% of carboxyl groups of the fragments were modified at a concentration of 0.05M carbodiimide reagent. The interaction of calcium and local anesthetics such as procaine and lidocaine with these fragments still showed typical competition. However, when the calcium binding was decreased to 8% at a higher concentration of carbodiimide reagent (0.08M), the local anesthetics still inhibited the calcium binding, but were not competitive in nature. In other words, if concentrations of the carbodiimide reagent were raised, the degree of inhibition by the local anesthetics was gradually decreased and was not competitive in nature. Finally, no inhibition was demonstrated when the concentration of the reagent was 0.1 to 0.4M. The above findings, seem to suggest that local anesthetics such as procaine and lidocaine interact with carboxyl groups, in addition to phosphodiester groups of phospholipids as previously reported, and inhibited competitively calcium binding to carboxyl groups of the membrane fragments.
Anesthetics, Local/pharmacology* ; Calcium/metabolism* ; Carbodiimides/pharmacology* ; Cell Membrane/metabolism ; Erythrocytes/metabolism* ; Human ; In Vitro ; Protein Binding

To investigate the changes of free hemoglobin (FHb) content after mixing type B whole blood with different amounts of type O whole blood at room temperature and at 37 degrees C, two lots of type B whole blood stored at 4 degrees C for 24 hours were randomly taken as recipient blood, and were packed as 60 ml respectively. Type O blood was taken as donor blood. 60 ml type B whole bloods were mixed with different amounts of type O whole blood, i.e. with 9, 12, 15 and 18 ml. The mixed blood was packed into 100 ml plastic blood bags and stored at 37 degrees C or room temperature, shaken once every 15 minutes. Free hemoglobin content was determined for the harvested samples at 1, 2, 4, 8 and 12 hours after store. The results showed that there was no significant elevation of FHb within 12 hours after mixing B whole blood with different amounts of type O whole blood. In another lot, there was obvious difference in FHb after 1 hour store along with the prolongation of store at either room temperature or 37 degrees C. In one lot, there was no difference of FHb (P > 0.05) during 1 - 8 hours of store at room temperature or 37 degrees C, but significant difference at 12 hours of store (P < 0.001). In another lot, there was no difference of FHb (P > 0.05) within 1 hour of store at room temperature and at 37 degrees C, but significant difference during 2 approximately 8 hours of store (P < 0.001). It is concluded that the FHb would not change significantly within 12 hours after type B blood was mixed with 1 200 ml of type O whole blood, but when the mixed blood was placed at room temperature or at 37 degrees C for 8 hours, the FHb content approaches, even exceeds 170.4 mg/L which was observed in the blood stored for 2 days. It suggests that freshly collected blood must be put into refrigerator of 2 approximately 4 degrees C for storing as soon as possible, so as to decrease the catabolism of erythrocyte and the releasing of FHb and other metabolites which are deleterious to the recipients.
ABO Blood-Group System ; metabolism ; Blood Preservation ; methods ; Erythrocytes ; metabolism ; Hemoglobins ; metabolism ; Humans ; Time Factors

Erythrocytes are devoid of nuclei and mitochondria which are the crucial elements of apoptosis, so their programmed suicidal death is called eryptosis. Eryptosis is characterized by cell shrinkage, membrane blebbing, activation of proteases, and phosphatidylserine exposure. Prostaglandin E(2) (PGE(2)) activates nonselective cation channels that increase cytosolic Ca(2+) activity and platelet-activating factor (PAF) activates a sphingomyelinase which lead to formation of ceramide. Either can lead to membrane scrambling with subsequent phosphatidylserine exposure. Exposed phosphatidylserine is recognized by macrophages that engulf and degrade the injured cells. As such, eryptosis can clear the injured red blood cells and avoid the release of hemoglobin. The signaling of eryptosis includes PGE(2), cation channels, PAF, ceramide, protein kinase C, and in some instances, caspases. In this review, the PGE(2), PAF and protein kinase pathways, erythrocyte surface receptor-mediated effects, oxidative stress and caspase effects, the inhibitory factors of eryptosis and the clinical eryptosis-related diseases are discussed.
Apoptosis ; physiology ; Dinoprostone ; metabolism ; Erythrocytes ; metabolism ; physiology ; Humans ; Platelet Activating Factor ; metabolism ; Signal Transduction

Erythrocytes ; metabolism ; Humans ; Kidney Diseases ; genetics ; metabolism ; Mitochondria ; genetics ; Mitochondrial Membranes ; metabolism ; Oxidative Stress ; genetics ; physiology

We investigated the expression of aquaporin 1 (AQP1) in tissues from canines with an inherited anomaly that causes their erythrocytes to have high K+. Northern blot analysis revealed abundant AQP1 expression in lung and kidney, though little expression was found in spleen. Using anti-C-terminus for dog AQP1, abundant expression was shown in kidney, trachea, and eye, but little expression was shown in pancreas and cerebrum, indicating that AQP1 expression in canine tissues is similar to that noted in other mammals.
Animals ; Aquaporin 1/*metabolism ; Blotting, Northern ; Dogs ; Erythrocytes/*chemistry ; Immunoblotting ; Potassium/*analysis ; Viscera/metabolism

This study was purposed to establish the method of quantifying RhD antigen on red blood cells (RBC) by flow cytometry (FCM) and to explore the expression of D antigen on RBC of different RhD serotype. RhD(+) RBCs and RhD(-) RBCs were mixed in 1:1 ratio. Cells were stained by the indirect method (IgG anti-D as the first antibody, FITC-anti-IgG F(ab')2 as the second antibody), and the ratio of RhD(+) on RBCs was quantified by FCM. The optimal dosage of IgG anti-D was defined. Expression of RhD antigen on RBC of RhD(+), weak D, RhDel and RhD(-) type were detected by FCM. The results showed that optimal dilution of IgG anti-D monoclonal antibody was 1:4, 1x10(6) cells/50 microl. The percentage of D(+) RBC of RhD(+), weak D, RhDel and RhD(-) type were 96.8+/-2.97%, 79.5+/-9.88%, 47.8+/-11.43%, 3.7+/-2.96%, respectively. The mean fluorescence intensity (MFI) of RhD antigen expression of RhD(+), weak D, RhDel and RhD(-) type were 33.3+/-6.21 Dal, 18.6+/-5.39 Dal, 7.10+/-1.17 Dal, 0.79+/-0.55 Dal, respectively. In conclusion, there are significant differences of RhD antigen expressions among RBC of different RhD serotypes. The level of antigen on RhD(+) RBC is the highest and then weak D the next, while the level of antigen on RhDel RBC is the lowest level.
Blood Donors ; Erythrocytes ; immunology ; metabolism ; Flow Cytometry ; methods ; Humans ; Rh-Hr Blood-Group System ; immunology ; metabolism

Acetylcholinesterase ; metabolism ; Adult ; Erythrocyte Count ; Erythrocytes ; enzymology ; Humans ; Male ; Motion Sickness ; metabolism ; physiopathology ; Naval Medicine