Objective: To report gene mutations in nine patients with hereditary elliptocytosis (HE) and analyze the characteristics of pathogenic gene mutations in HE. Methods: The clinical and gene mutations of nine patients clinically diagnosed with HE at Institute of Hematology & Blood Diseases Hospital from June 2018 to February 2022 were reported and verified by next-generation sequencing to analyze the relationship between gene mutations and clinical phenotypes. Results: Erythrocyte membrane protein gene mutations were detected among nine patients with HE, including six with SPTA1 mutation, one with SPTB mutation, one with EPB41 mutation, and one with chromosome 20 copy deletion. A total of 11 gene mutation sites were involved, including 6 known mutations and 5 novel mutations. The five novel mutations included SPTA1: c.1247A>C (p. K416T) in exon 9, c.1891delG (p. A631fs*17) in exon 15, E6-E12 Del; SPTB: c.154C>T (p. R52W) ; and EPB41: c.1636A>G (p. I546V) . Three of the six patients with the SPTA1 mutation were SPTA1 exon 9 mutation. Conclusion: SPTA1 is the most common mutant gene in patients with HE.
Humans ; Mutation ; Elliptocytosis, Hereditary/metabolism* ; Erythrocyte Membrane/metabolism* ; Exons ; High-Throughput Nucleotide Sequencing ; Spherocytosis, Hereditary/metabolism*

OBJECTIVETo study the toxicity of methomyl to acetylcholinesterase (AChE) in different regions.

METHODSThe optimal temperature and time for measurement of AChE activity were determined in vitro. The dose- and time-response relationships of methomyl with AChE activity in human erythrocyte membrane, rat erythrocyte membrane, cortical synapses, cerebellar synapses, hippocampal synapses, and striatal synapses were evaluated. The half maximal inhibitory concentration (IC50) and bimolecular rate constant (K) of methomyl for AChE activity in different regions were calculated, and the type of inhibition of AChE activity by methomyl was determined.

RESULTSAChE achieved the maximum activity at 370 °C, and the optimal time to determine initial reaction velocity was 0-17 min. There were dose- and time-response relationships between methomyl and AChE activity in the erythrocyte membrane and various brain areas. The IC50 value of methomyl for AChE activity in human erythrocyte membrane was higher than that in rat erythrocyte membrane, while the Ki value of methomyl for AChE activity in rat erythrocyte membrane was higher than that in human erythrocyte membrane. Among synapses in various brain areas, the striatum had the highest IC50 value, followed by the cerebellum, cerebral cortex, and hippocampus, while the cerebral cortex had the highest Ki value, followed by the hippocampus, striatum, and cerebellum. Lineweaver-Burk diagram demonstrated that with increasing concentration of methomyl, the maximum reaction velocity (Vmax) of AChE decreased, and the Michaelis constant (Km) remained the same.

CONCLUSIONMethomyl is a reversible non-competitive inhibitor of AChE. AChE of rat erythrocyte membrane is more sensitive to methomyl than that of human erythrocyte membrane; the cerebral cortical synapses have the most sensitive AChE to methomyl among synapses in various brain areas.

Acetylcholinesterase ; metabolism ; Animals ; Cerebellum ; drug effects ; Cerebral Cortex ; drug effects ; Erythrocyte Membrane ; drug effects ; enzymology ; Hippocampus ; drug effects ; Humans ; Inhibitory Concentration 50 ; Methomyl ; toxicity ; Rats ; Synapses ; drug effects ; Toxicity Tests

PURPOSE: The aim of this study was to determine whether plasma lecithin:cholesterol acyltransferase (pLCAT) and erythrocyte membrane Na(+)-K(+)-ATPase ase (emNaKATPs) activity have a correlation in breast cancer. This study compared these parameters at time points before and after treatment with radiotherapy. METHODS: The levels of pLCAT and emNaKATPs were assessed in 30 patients with breast carcinoma and 20 control subjects. While emNaKATPs was measured with spectrophotometric method, pLCAT levels was measured using a specific enzyme-linked immunosorbent assay. RESULTS: pLCAT levels, both before and after radiotherapy, were found to be decreased in breast cancer patients than in the controls groups (p<0.001 and p<0.001, respectively). Also, pLCAT levels after radiotherapy were found to be decreased in breast cancer patients than the pLCAT levels before radiotherapy (p<0.001). The emNaKATPs activity were higher in the control group than in the breast cancer patients before/after radiotherapy (RT) (p<0.001 and p<0.001, respectively). At the same time, emNaKATPs activity before RT was higher in the breast cancer patients than emNaKATPs activity after RT (p<0.001). There was a significant correlation between pLCAT and emNaKATPs activity in breast cancer patients receiving radiotherapy (r=0.63, p<0.001), but no correlation between in breast cancer patients before RT and control group (r=0.023, p>0.05). CONCLUSION: The results of the present study demonstrated that decreased pLCAT and emNaKATPs activity levels in breast cancer patients after/before RT than control group. In addition, decreased emNaKATPs activity in breast cancer patients receiving radiotherapy may be due to decreased pLCAT concentrations and RT beam. In our opinion, altered activities of pLCAT and emNaKATPs are linked to the treatment effect of radiotherapy. These data may clarify the development of cell membrane dysfunction and lipid metabolism in breast cancer patients receiving radiotherapy.
Breast ; Breast Neoplasms ; Cell Membrane ; Cholesterol ; Erythrocyte Membrane ; Humans ; Lecithins ; Lipid Metabolism ; Plasma ; Sterol O-Acyltransferase

OBJECTIVETo investigate the effect of Yisui Shengxue Granule (, YSSXG), a complex Chinese medicine, on the oxidative damage of erythrocytes from patients with hemoglobin H (HbH) disease.

METHODSTwenty-two patients with HbH disease and 22 healthy volunteers were observed. YSSXG was given to patients with HbH disease for 3 months. Before and after the 3-month treatment, blood parameters [hemoglobin (Hb), red blood cells (RBCs), and reticulocyte percent (Ret)] were examined; inclusion bodies in erythrocytes were observed by transmission electron microscopy (TEM); activities of antioxidant defense enzymes [superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (Cat)] and erythrocyte membrane malondialdehyde (MDA) concentrations were determined.

RESULTSIn patients with HbH disease, measured values of RBC and Hb obtained from the first to the third months after treatment with YSSXG were significantly higher than before treatment (P<0.01). Measured values of Ret from the second to the third months after treatment were significantly lower than before treatment (P<0.05 and P<0.01, respectively). Prior to treatment with YSSXG, TEM images of RBCs showed the presence of numerous inclusion bodies. After treatment with YSSXG, the amount and volume of inclusion bodies decreased. Treatment with YSSXG also led to a significant increase in SOD activity (P<0.01), a decrease in Cat activity (P<0.01), and no significant differences in GSHPx activity (P>0.05) or MDA concentration (P>0.05). However, compared with the healthy counterparts, SOD, GSH-Px, and Cat activities presented at high levels (P<0.01) both before and after treatment.

CONCLUSIONSYSSXG could improve the degree of hemolysis and anemia in patients with HbH disease. The mechanism may be related to its antioxidative effects, which could elevate the activity of total SOD in erythrocytes and efficiently inhibit the oxidative precipitation of β-globin chains.

Adolescent ; Adult ; Catalase ; metabolism ; Child ; Child, Preschool ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Erythrocyte Membrane ; drug effects ; metabolism ; ultrastructure ; Erythrocytes ; drug effects ; enzymology ; pathology ; ultrastructure ; Female ; Glutathione Peroxidase ; metabolism ; Humans ; Inclusion Bodies ; drug effects ; ultrastructure ; Male ; Malondialdehyde ; metabolism ; Oxidative Stress ; drug effects ; Superoxide Dismutase ; metabolism ; Young Adult ; alpha-Thalassemia ; blood ; drug therapy ; pathology

OBJECTIVETo investigate the relationships between erythrocyte membrane n-6:n-3 PUFAs ratio and blood lipids and high sensitivity C-reactive protein (hs-CRP).

METHODSThe observational study consisted of a population-based cross-sectional study of 456 Chinese and a subsequent 1-year follow-up study of 171 subjects with the fasting plasma total cholesterol of 5.13-8.00 mmol/L.

RESULTSIn 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 (Δ=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).

CONCLUSIONOur results suggested that the balance between n-6 and n-3 fatty acids may optimize the cardiovascular benefits from dietary PUFAs.

C-Reactive Protein ; metabolism ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Cross-Sectional Studies ; Erythrocyte Membrane ; metabolism ; Fatty Acids, Omega-3 ; blood ; Fatty Acids, Omega-6 ; blood ; Female ; Humans ; Lipids ; blood ; Male ; Middle Aged ; Triglycerides ; blood

OBJECTIVETo observe the changes of erythrocyte deformability in rats acclimatized to hypoxia and its molemechanism.

METHODSMale rats were randomly divided into three groups (n = 10): normal control group, acute hypoxia group and hypoxia acclimatization group. Animals were exposed to hypoxia for 0, 1, 28 d, blooded from their hearts after anaesthetized, respectively. Erythrocyte deformability, membrane fluidity, cholesterin and total lipid, lipid components of erythrocyte membrane, erythrocyte membrane ATPase and the concentrations of Na+ and Ca2+ were measured respectively. The two-dimensional electrophoresis maps of the rats erythrocyte membrane protein were achieved. The different protein spots were founded by image master 2D elite and identified by mass spectrum.

RESULTS(1) In acute hypoxia group, the deformability, membrane fluidity, the content of membrane cholesterin and total lipid were declined. The content of phosphatidylserines (PS), sphingomyelin (SM) in erythrocyte membrane lipids were increased, phosphatidylcholine (PC) reduced. The activity of ATP enzymes reduced and the concentration of Na+ and Ca2+ in erythrocyte increased. The two-dimensional electrophoresis maps of the rats erythrocyte membrane protein were achieved. Four of the seven protein spots selected increased and three of them showed no change. (2) In hypoxia acclimatization group, the deformability, membrane fluidity, the content of membrane cholesterin and total lipid were increased than those in acute hypoxia group, similar to normal group. The content of PS, SM in erythrocyte membrane lipids were reduced, PC increased. The activity of ATP enzymes induced and the concentration of Na+ and Ca2+ in erythrocyte increased after hypoxia acclimatization. Four of those protein spots mentioned increase and three declined after hypoxia acclimatization. They were respectively proved by mass spectrum to be alexin binding protein, aquaporin chip, membrane inhibitor reactive lysis, phospholipids scramblase, glucose transferase, aminophospholipid translocases, ATP-dependent floppase, the latter three proteins were associate with the overturning of erythrocyte membrane lipids.

CONCLUSIONAcute hypoxia caused the corresponding damage of erythrocyte deformability, erythrocyte membrane fluidity, erythrocyte membrane proteins erythrocyte expression, the activity of membrane ATPase and the concentration of Na+ and Ca2+ in erythrocyte. The parameters above were improved after hypoxia acclimatization, so hypoxia acclimatization effected positively in the damage to erythrocyte due to acute hypoxia. The three membrane proteins might play important roles in the deformability improved by hypoxia acclimatization, which included phospholipids scramblase, aminophospholipid translocases and ATP-dependent floppase.

Acclimatization ; physiology ; Adenosine Triphosphatases ; metabolism ; Altitude ; Animals ; Calcium ; metabolism ; Erythrocyte Deformability ; physiology ; Erythrocyte Membrane ; metabolism ; Hypoxia ; blood ; physiopathology ; Male ; Membrane Fluidity ; Phospholipid Transfer Proteins ; metabolism ; Rats ; Sodium ; metabolism

OBJECTIVETo study the effects of hermap gene on kinases in erythroid signal transduction pathway and investigate the mechanism of hermap on erythroid differentiation.

METHODSThe K562 cells expressing hermap and hermap-siRNA respectively were established for up- and down-regulating the expression of hermap gene. These K562 cells were then induced by Ara-C to erythroid differentiation and analyzed at 0, 24, 48, 72 and 96 h, respectively, for cell morphology and biphenylamine staining positive cells, determination of CD235a, CD36, kinases p-STAT5, p-Akt, p-MAPK and p-c-JUN by FCM; and quantification of hermap gene and γ (Aγ,Gγ) globin gene by FQ-PCR.

RESULTSWith up-regulating hermap gene and inducing by Ara-C, K562 cells were changing to low ratio of nucleus to cytoplasm, cytoplasm colour from basophilic to pinkish or amethyst tinge, increase of number of biphenylamine positive cells and expression of CD235a, CD36, γ (Aγ,Gγ) globin gene, hermap gene and p-STAT5 from 0 to 96 h. At 0, 24, 48, 72 and 96 h of culture, the positive rates of p-STAT5 cells were detected of 0.46%, 4.54%, 20.01%, 23.65% and 33.08%, respectively. This results demonstrated that there was a positive correlation between expression of p-STAT5 and hermap gene expression (P < 0.05).

CONCLUSIONhermap gene can stimulate erythroid differentiation of Ara-C induced K562 cells mainly through JAK/STAT5 signal transduction pathway.

Cell Differentiation ; Erythrocyte Membrane ; Erythrocytes ; cytology ; Erythropoiesis ; Gene Expression ; Humans ; K562 Cells ; Receptors, Erythropoietin ; genetics ; STAT5 Transcription Factor ; metabolism ; Signal Transduction

Acute Coronary Syndrome ; metabolism ; pathology ; physiopathology ; Cholesterol ; metabolism ; Erythrocyte Membrane ; metabolism ; Humans

Adult ; Calcium-Transporting ATPases ; metabolism ; Erythrocyte Membrane ; metabolism ; Female ; Humans ; Hydrocarbons ; adverse effects ; Lipid Peroxidation ; Male ; Middle Aged ; Occupational Exposure ; Sodium-Potassium-Exchanging ATPase ; metabolism ; Young Adult

Aged ; Anion Exchange Protein 1, Erythrocyte ; metabolism ; Brenner Tumor ; metabolism ; pathology ; surgery ; Carcinoma, Transitional Cell ; metabolism ; pathology ; surgery ; Chromogranin A ; metabolism ; Cystectomy ; Female ; Humans ; Hysterectomy ; Membrane Proteins ; metabolism ; Neoplasms, Multiple Primary ; metabolism ; pathology ; surgery ; Ovarian Neoplasms ; metabolism ; pathology ; surgery ; Urinary Bladder Neoplasms ; metabolism ; pathology ; surgery