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

OBJECTIVETo investigate whether miR-492 is involved in the post-transcriptional regulation of OK blood group antigen expression on red blood cells.

METHODSTwo 3'-UTR fragments of the BSG gene were synthesized with a chemical method, which respectively encompassed the BSG rs8259 TT or BSG rs8259 AA sites. The fragments were added with Xho I and Not I restriction enzyme cutting sites at both ends and cloned into a pUC57 vector, which in turn was constructed into a psiCHECK-2 vector and verified by sequencing. K562 cells were transfected with various combinations of miR-492 mimic and constructed psiCHECK2-BSG-T or psiCHECK2-BSG-A recombinant plasmid. A blank control group was set up. Each transfection experiment was repeated three times. The activity of Renilla reniformis luciferase was determined and normalized with that of firefly luciferase, and detected with a dual-luciferase reporter assay system. The data were subjected to statistical analysis.

RESULTSThe sequencing results confirmed that the recombinant psiCHECK2 plasmids containing the BSG rs8259 TT or rs8259 AA sites were constructed successfully. The results of dual-luciferase report gene detection showed that the miR-492 mimic could significantly inhibit psiCHECK2-BSG-T at a concentration over 100 nmol/L. However, it could not inhibit psiCHECK-BSG-A.

CONCLUSIONmiR-492 may be involved in the regulation of OK antigen expression on red blood cells with the BSG rs8259 TT genotype.

Basigin ; genetics ; Blood Group Antigens ; genetics ; Erythrocytes ; immunology ; Gene Expression Regulation ; Genotype ; Humans ; MicroRNAs ; physiology

Amphibian skin contains rich bioactive peptides. Especially, a large amount of antimicrobial peptides have been identified from amphibian skin secretions. Antimicrobial peptides display potent cytolytic activities against a range of pathogenic bacteria and fungi and play important defense roles. No antimicrobial peptides have been reported from toads belonging to the family of Pelobatidae. In this work, two novel antimicrobial peptides (Megin 1 and Megin 2) were purified and characterized from the skin venoms of spadefoot toad Megophrys minor (Pelobatidae, Anura, Amphibia). Megin 1 had an amino acid sequence of FLKGCWTKWYSLKPKCPF-NH2, which was composed of 18 amino acid residues and contained an intra-molecular disulfide bridge and an amidated C-terminus. Megin 2 had an amino acid sequence of FFVLKFLLKWAGKVGLEHLACKFKNWC, which was composed of 27 amino acid residues and contained an intra-molecular disulfide bridge. Both Megin 1 and Megin 2 showed potential antimicrobial abilities against bacteria and fungi. The MICs of Megin 1 against Escherichia coli, Bacillus dysenteriae, Staphylococcus aureus, Bacillus subtilis, and Candida albicans were 25, 3, 6.25, 3, and 50 μg·mL(-1), respectively. The corresponding MICs for Megin 2 were 6.25, 1.5, 12.5, 1.5, and 12.5 μg·mL(-1), respectively. They also exerted strong hemolytic activity against human and rabbit red cells. The results suggested that megin peptides in the toad skin of M. minor displayed toxic effects on both eukaryotes and prokaryotes. This was the first report of antimicrobial peptides from amphibians belonging to the family of Pelobatidae.
Amino Acid Sequence ; Amphibian Venoms ; chemistry ; immunology ; isolation & purification ; Animals ; Anura ; immunology ; Bacillus ; Candida albicans ; Erythrocytes ; physiology ; Escherichia coli ; Female ; Hemolysis ; Humans ; Male ; Peptides ; chemistry ; immunology ; isolation & purification ; Rabbits ; Sequence Alignment ; Skin ; chemistry ; immunology ; Staphylococcus aureus

To demonstrate the effect of AB serum on terminal erythroid differentiation ex vivo.
 Methods: After separation of CD34+ cells from cord blood, the cells were cultured and divided into a control group and an experimental group. The effects of AB serum were examined by the expressions of different markers (GPA, Band3 and α4-integrin) for erythroblast differentiation and enucleation by flow cytometry. 
 Results: The CD34+ cells were successfully differentiated to enucleated red blood cells. There were evident differences among the expressions of GPA, Band3 and α4-integrin between the 2 groups. The percentage of GPA positive cells in the experimental group was bigger than that in the control group in every time point. The expression of Band3 in the experimental group was higher than that in the control group. The expression of α4-integrin in the experimental group was lower than that in the control group. In addition, the enucleation rate in the experimental group was higher than that in the control group.
 Conclusion: AB serum can promote the cell differentiation and enucleation during terminal erythroid differentiation in vitro.
ABO Blood-Group System ; blood ; physiology ; Anion Exchange Protein 1, Erythrocyte ; metabolism ; Antigens, CD34 ; blood ; Cell Differentiation ; genetics ; physiology ; Cell Nucleus ; Cells, Cultured ; Erythrocytes ; physiology ; ultrastructure ; Erythropoiesis ; genetics ; physiology ; Fetal Blood ; cytology ; physiology ; Flow Cytometry ; Glycophorins ; metabolism ; Humans ; Integrin alpha4beta1 ; metabolism

OBJECTIVEBlood transfusion saves lives but may also increase the risk of injury. The objective of this review was to evaluate the possible adverse effects related to transfusion of red blood cell (RBC) concentrates stored for prolonged periods.

DATA SOURCESThe data used in this review were mainly from PubMed articles published in English up to February 2015.

STUDY SELECTIONClinical and basic research articles were selected according to their relevance to this topic.

RESULTSThe ex vivo changes to RBC that occur during storage are collectively called storage lesion. It is still inconclusive if transfusion of RBC with storage lesion has clinical relevance. Multiple ongoing prospective randomized controlled trials are aimed to clarify this clinical issue. It was observed that the adverse events related to stored RBC transfusion were prominent in certain patient populations, including trauma, critical care, pediatric, and cardiac surgery patients, which leads to the investigation of underlying mechanisms. It is demonstrated that free hemoglobin toxicity, decreasing of nitric oxide bioavailability, and free iron-induced increasing of inflammation may play an important role in this process.

CONCLUSIONIt is still unclear whether transfusion of older RBC has adverse effects, and if so, which factors determine such clinical effects. However, considering the magnitude of transfusion and the widespread medical significance, potential preventive strategies should be considered, especially for the susceptible recipients.

Blood Preservation ; adverse effects ; Erythrocyte Transfusion ; adverse effects ; Erythrocytes ; metabolism ; physiology ; Humans ; Nitric Oxide ; metabolism ; Time Factors

BACKGROUNDBone marrow hematopoietic function suppression is one of the most common side effects of chemotherapy. After chemotherapy, the bone marrow structure gets destroyed and the cells died, which might cause the hematopoietic function suppression. Heme oxygenase-1 (HO-1) is a key enzyme of antioxidative metabolism that associates with cell proliferation and resistance to apoptosis. The aim of this study was to restore or resist the bone marrow from the damage of chemotherapy by the HO-1 expression of mouse mesenchymal stem cells (mMSCs) homing to the mice which had the chemotherapy-induced bone marrow suppression.

METHODSOne hundred and sixty female Balb/c mice (6-8-weeks old) were randomly divided into four groups. Each group was performed in 40 mice. The control group was intraperitoneally injected for 5 days and tail intravenously injected on the 6th day with normal saline. The chemotherapy-induced bone marrow suppression was established by intraperitoneally injecting cyclophosphamide (CTX) into the mice which performed as the chemotherapy group. The mMSCs were tail intravenously injected into 40 chemotherapically damaged mice which served as the mMSCs group. The difference between the HO-1 group and the mMSCs group was the injected cells. The HO-1 group was tail intravenously injected into the mMSCs that highly expressed HO-1 which was stimulated by hemin. The expression of HO-1 was analyzed by Western blotting and RT-PCR. Cell proliferation was measured using the 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay. Histopathologic examinations were performed 1 week after injection.

RESULTSCompared with the control group, the expression levels of HO-1 mRNA and protein were significantly higher in the HO-1 group (all P < 0.05), even obviously than the mMSCs group. CTX treatment induced apoptosis and inhibited proliferation. After injected, the white blood cell (WBC), red blood cell (RBC) and platelet (PLT) declined fast and down to the bottom at the 7th day. The bone marrow structure was destroyed incomplete. In vitro, the survival rate of cells in chemotherapy group was less than 50% after 24 hours. In contrast, mMSCs could do a favor to the cellular cleavage and proliferation. They slowed down the cell mortality and more than 50% cells survived after 24 hours. The effects of blocking apoptosis and bone marrow recovery could be more effective in the HO-1 group. In the HO-1 group, it had observed that the bone marrow structure became complete and the hemogram closed to normal at 7th day.

CONCLUSIONSHO-1 played an important role in promoting the recovery of CTX-induced hematopoietic damage. We suggest that HO-1 is able to restore the functions of chemotherapy-induced hematopoietic damage.

Animals ; Apoptosis ; drug effects ; Blood Platelets ; drug effects ; Blotting, Western ; Bone Marrow ; drug effects ; enzymology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cyclophosphamide ; toxicity ; Erythrocytes ; drug effects ; Female ; Heme Oxygenase-1 ; genetics ; metabolism ; Leukocytes ; drug effects ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; enzymology ; physiology ; Mice ; Mice, Inbred BALB C ; Reverse Transcriptase Polymerase Chain Reaction

Erythrocytes lack nuclei and mitochondria, critical elements in the machinery of nucleated cell apoptosis. However, most recently, it became obvious that erythrocytes may undergo programmed aging, as well as suicidal death. The term eryptosis has been coined to describe the suicidal erythrocyte death. Eryptosis is triggered mainly by increased cytosolic Ca(2+) activity, in turn, Ca(2+) activates Ca(2+)-sensitive K(+) channels, scramblase, calpain and other proteases, respectively. A series of molecular events of erythrocyte programmed death induced. The cascade reaction of related molecules and finally lead to cell clearance. There is evidence suggesting that erythrocytes aging and death process are regulated tightly and there are many molecular participants and signaling pathways involved in aging and death process of erythrocytes. Erythrocytes have already been used as a model for aging study, and the knowledge about mechanisms involved in eryptosis may provide an important clue to understand the mechanisms involved in suicidal death of nucleated cells. In this review the factors influencing programmed death of erythrocytes, the role of Ca(2+) and ceramide in programmed death of erythrocytes, the role of blebbing in process of erythrocyte aging, the antigens of erythrocyte aging and so on are summarized.
Calcium ; physiology ; Cell Death ; Cellular Senescence ; Ceramides ; physiology ; Erythrocytes ; cytology ; Humans

OBJECTIVETo observe the change of erythrocyte theology in rabbits with acute renal failure (ARF).

METHODSThirty-eight healthy rabbits were randomly divided into control group (n = 8), model group (establishing ARF model via intramuscular injection 1% HgCl2, and divided into 12 h, 24 h, 48 h subgroups, all n = 10), the arterial blood sample was taken out through carotid artery at corresponding times after anesthetization with urethane, for detecting the indices of renal function and erythrocyte rheology.

RESULTSThe levels of urea and creatinine in plasma of model rabbits at 12 h, 24 h and 48 h were higher than those of control group, and there was a rise trend along with the time extension. The erythrocytes electrophoresis time at 12 h of model group was higher, the electrophoresis rate and migration rate of erythrocytes were lower compared with those of control group; the erythrocytes electrophoresis time at 24 h of model group was lower and the electrophoresis rate and migration rate were higher compared with those of model group at 12 h; and there were no statistical differences in erythrocytes electrophoresis indices between model group at 48 h and other groups. Meanwhile, there was a rise trend in erythrocyte sedimentation rate (ESR), K value of equation and emendation along with the time extension of ARF, but these indices only at 48 h of model group was lower significantly than that of control group. There were no statistical differences in aggregation index and deformability index of erythrocytes among groups.

CONCLUSIONDuring the process of ARF, the erythrocytes electrophoresis time lengthen and electrophoresis rate and migration rate decrease at early stage, and these indices gradually return to normal; the indices of ESR increase gradually.

Acute Kidney Injury ; blood ; physiopathology ; Animals ; Erythrocyte Indices ; Erythrocytes ; physiology ; Hemorheology ; Rabbits

We recently encountered a case of hereditary spherocytosis coexisting with Gilbert's syndrome. Patient was initially diagnosed with Gilbert's syndrome and observed, but other findings suggestive of concurrent hemolysis, such as splenomegaly and gallstones were noted during the follow-up period. Therefore, further evaluations, including a peripheral blood smear, osmotic fragility test, autohemolysis test, and red blood cell membrane protein test were performed, and coexisting hereditary spherocytosis was diagnosed. Genotyping of the conjugation enzyme uridine diphosphate-glucuronosyltransferase was used to confirm Gilbert's syndrome. Because of the high prevalence rates and similar symptoms of these 2 diseases, hereditary spherocytosis can be masked in patients with Gilbert's syndrome. In review of a case and other article, the possibility of the coexistence of these 2 diseases should be considered, especially in patients with unconjugated hyperbilirubinemia who also have splenomegaly and gallstones.
Adult ; Erythrocytes/physiology ; Gallstones/etiology ; Genotype ; Gilbert Disease/complications/*diagnosis/genetics ; Glucuronosyltransferase/genetics ; Hemolysis ; Humans ; Hyperbilirubinemia/etiology ; Male ; Polymorphism, Single Nucleotide ; Spherocytosis, Hereditary/complications/*diagnosis/genetics ; Splenomegaly/etiology

OBJECTIVETo observe the changes of blood viscosity and erythrocyte rheology in mice after acute hypoxic hypoxia (AHH).

METHODSThirty-two Kui-ming mice were randomly divided into control group, AHH group (duplicating AHH model, and divided into 5 min, 8 min, 11 min subgroups), the blood sample was taken out from heart after neck dislocation at corresponding times, for detecting the blood viscosity and erythrocyte rheology indices.

RESULTSCompared with control group, the whole blood viscosity at different shears, whole blood reduced viscosity, whole blood relative viscosity were lower and the erythrocytes aggregation index was higher in AHH 5 min group; the electrophoresis time was longer and the electrophoresis length, migration of erythrocyte were lower in AHH 8 min and AHH 11 min groups. The whole blood reduced viscosity, whole blood relative viscosity, erythrocytes aggregation index in AHH 8 min group were higher, and the erythrocyte deformability index was lower significantly than that of AHH 5 min group, respectively.

CONCLUSIONThese data suggested that the AHH could induce the blood viscosity and electrophoresis ability.

Animals ; Blood Viscosity ; Erythrocyte Aggregation ; Erythrocyte Deformability ; Erythrocyte Indices ; Erythrocytes ; physiology ; Hypoxia ; blood ; etiology ; Male ; Mice ; Mice, Inbred Strains