OBJECTIVETo study the clinical features of two families with distal renal tubular acidosis (dRTA) and mutations in the pathogenic gene SLC4A1.

METHODSFamily investigation, medical history collection, and measurement of biochemical parameters were performed to analyze the clinical phenotype and genetic characteristics of dRTA. Direct sequencing was used to detect SLC4A1 gene mutations.

RESULTSThree patients in these two families (two of them were mother and son) were diagnosed with dRTA with typical clinical features, including short stature, metabolic acidosis, alkaline urine, hypokalemia, and nephrocalcinosis. SLC4A1 gene analysis showed that all the three patients had a pathogenic missense mutation R589H (c.1766G>A). The child in family 1 had a de novo mutation of SLC4A1, and the child in family 2 had an SLC4A1 gene mutation inherited from the mother, which met the characteristic of autosomal dominant inheritance.

CONCLUSIONSThis study reports the R589H mutation in SLC4A1 gene in families with hereditary dRTA for the first time in China. Clinical physicians should perform gene detection for patients suspected of hereditary dRTA to improve the diagnosis and treatment of this disease.

Acidosis, Renal Tubular ; genetics ; Anion Exchange Protein 1, Erythrocyte ; genetics ; Child ; Humans ; Male ; Mutation

BACKGROUND: The eosin-5'-maleimide (EMA) binding test using flow cytometry is a common method to measure reduced mean channel fluorescence (MCF) of EMA-labeled red blood cells (RBCs) from patients with red cell membrane disorders. The basic principle of the EMA-RBC binding test involves the covalent binding of EMA to lysine-430 on the first extracellular loop of band 3 protein. METHODS: In the present study, the MCF of EMA was analyzed for samples derived from 12 healthy volunteers (controls) to determine the stability (i.e., the percentage decrease in fluorescence) of EMA over a period of 1 year. RESULTS: Comparison of periodical MCF readings over time, that is, at 2-month intervals, showed that there were no significant changes in mean channel fluorescence for up to 6 months; however, there was a significant decrease in MCF at 8 months. CONCLUSION: For optimal dye utilization, EMA remained stable only for up to 6 months. Therefore, we recommend reconstitution of the dye every 6 months when implementing this test and storage at -80degrees C in dark conditions.
Anion Exchange Protein 1, Erythrocyte ; Cell Membrane* ; Erythrocytes ; Flow Cytometry ; Fluorescence ; Healthy Volunteers ; Humans ; Reading

AIM AND METHODSThe purpose of this investigation was to determine the changes of erythrocyte deformability, band-3 protein and actin in the definite volume of erythrocyte membrane after high intensity running training and recovery in rats.

RESULTSLong-term training could significantly improve erythrocyte deformability and the quantity of membrane proteins. Erythrocyte deformability, band-3 protein and actin decreased transitorily at varying degrees after inadaptable high intensity exercise. One and two week training could improve erythrocyte deformability, the quantity of band-3 protein and actin after recovery.

CONCLUSIONAlterations of erythrocyte membrane protein after high intensity training could cause the change in erythrocyte membrane structure and hence influenced erythrocyte deformability. That was maybe one of mechanisms of training effecting erythrocyte deformability.

Actins ; metabolism ; Animals ; Anion Exchange Protein 1, Erythrocyte ; metabolism ; Erythrocyte Deformability ; Erythrocyte Indices ; Erythrocyte Membrane ; chemistry ; metabolism ; Physical Conditioning, Animal ; Rats ; Rats, Sprague-Dawley

OBJECTIVE: To investigate the feasibility and clinical significance of high resolution melting(HRM) curve analysis to detect SLC4A1 gene D38A and K56E mutations in the patients with hereditary spherocytosis(HS). METHODS: Peripheral blood was collected from 23 cases of HS for routine tests and their genomic DNA was extracted by routine technique. Specific primers of mutation sites D38A and K56E of SLC4A1 gene were designed. The HRM method was used to analyze all the samples, and then the results of HRM were verified with DNA sequencing technology. RESULTS: Among 23 specimens of HS patients, 6 cases of heterozygous mutant gene were detected by HRM technology, including 3 cases of D38A mutation and 3 cases of K56E mutation, which were confirmed by DNA sequencing. CONCLUSION The HRM technology can correctly detect 2 common mutation sites including D38A and K56E in SLC4A1 gene in an efficient, fast, and reliable way, which not only can be used for clinical diagnosis, but also expected to be a new method for clinical researchers to define gene mutation spectrum in HS patients.
Anion Exchange Protein 1, Erythrocyte ; genetics ; Base Sequence ; DNA Mutational Analysis ; DNA Primers ; Heterozygote ; Humans ; Mutation ; Spherocytosis, Hereditary ; genetics

OBJECTIVETo explore the hemolytic mechanism of glucose-6-phosphate dehydrogenase (G6PD) deficient erythrocytes in the view of phosphorylation of membrane protein.

METHODSThe alternation of membrane protein phosphorylation and the effect of dithiothreitol (DTT) on protein phosphorylation were analysed by Western blot technique. The activity of phosphotyrosine phosphatase (PTPs) was determined by using p-nitrophenyl phosphate as substrate.

RESULTSTyrosine phosphorylation of band 3 protein was obviously enhanced in G6PD-deficient erythrocytes. The activity of PTPs was low compared to the normal erythrocytes. The level of phosphotyrosine in G6PD-deficient erythrocytes incubated with DTT was almost the same as in those without DTT. The results were consistent with the activity of PTPs.

CONCLUSIONSPTPs activity reduction and tyrosine phosphorylation enhancement induced by oxidation in G6PD deficiency play an important role in erythrocytes hemolysis. However, the alternation of thiol group is not the only factor affecting the activity of PTPs in G6PD-deficient erythrocytes.

Anion Exchange Protein 1, Erythrocyte ; metabolism ; Blotting, Western ; Erythrocyte Membrane ; metabolism ; Glucosephosphate Dehydrogenase Deficiency ; enzymology ; metabolism ; Humans ; Phosphorylation ; Protein Tyrosine Phosphatases ; metabolism ; Tyrosine ; metabolism

Hereditary spherocytosis (HS) is caused by mutations in the SPTA1, SPTB, ANK1, SLC4A1, and EPB42 genes, all of which encode erythrocyte membrane proteins. Mutations in SLC4A1, which encodes band 3 protein, have rarely been reported as the causative factor among Korean patients with HS. Here, we report two Korean patients with HS carrying mutations in SLC4A1. Patient 1 was a 3-year-old girl with unremarkable past and family histories and was evaluated for anemia that was detected after a complete blood count. She was suspected of having HS considering the spherocytosis of her peripheral blood smear, increased osmotic fragility, hemolytic features in blood chemistry tests, and splenomegaly. Sequence analysis revealed that the patient harbored a single heterozygous missense mutation, c.2278C>T (p.Arg760Trp) in exon 17 of SLC4A1. Patient 2 was a 23-year-old man who had a prior history of intermittent jaundice. Although the patient did not have anemia, a genetic test for HS was performed due to evidence of hemolytic features in the blood chemistry test, splenomegaly, and a family history of HS. The test confirmed a single heterozygous missense mutation, c.2423G>T (p.Arg808Leu) in exon 18 of SLC4A1.
Anemia ; Anion Exchange Protein 1, Erythrocyte ; Blood Cell Count ; Chemistry ; Child, Preschool ; Erythrocyte Membrane ; Exons ; Female ; Humans ; Jaundice ; Mutation, Missense ; Osmotic Fragility ; Sequence Analysis ; Splenomegaly ; Young Adult

Anion Exchange Protein 1, Erythrocyte ; metabolism ; Breast ; metabolism ; pathology ; surgery ; Cadherins ; metabolism ; Female ; Fibrocystic Breast Disease ; metabolism ; pathology ; surgery ; Follow-Up Studies ; Humans ; Middle Aged ; S100 Proteins ; metabolism

Anion Exchange Protein 1, Erythrocyte ; metabolism ; Antiporters ; metabolism ; Child ; Chordoma ; metabolism ; pathology ; radiotherapy ; surgery ; Female ; Follow-Up Studies ; Humans ; Skull Base Neoplasms ; metabolism ; pathology ; radiotherapy ; surgery ; Vimentin ; metabolism

In developing rat kidney, apoptosis plays an important role in the morphogenesis of renal papilla, and hyperosmolality is well known to be one of regulating factor in apoptosis. The aim of this study was to determine the effect of hypoosmolality induced furosemide in neonate on cell proliferation and apoptosis in renal papilla. One-dayold pups were given two times a day, at 12 hour intervals, subcutaneous injection of furosemide (10 mg/kg BW) or saline for 4 days or 7days. We identified thick ascending limb by labeling with antibody to BSC1 (bumetanide-sensitive Na/K/2Cl cotransporter) and type A intercalated cell in collecting duct by labeling with antibody to band 3 protein. We also inspected apoptosis with TdT-mediated dUTP nick end labeling (TUNEL) method and cell proliferation with immunostaining for proliferating cell nuclear antigen (PCNA). The effect of hypoosmolality induced furosemide on neonatal rat kidney. 1) In furosemide-treated animals, body and kidney weights were reduced compare to control groups. the length of renal papillae of furosemide-treated groups were shortened compare to control groups. 2) Transformation from a cuboidal epithelium of thick ascending limb to a squamous epithelium of ascending thin limb in renal papilla was delayed in furosemide-treated groups. 3) In the inner medullary collecting duct of furosemide-treated groups, elimination of type A intercalated cells was delayed compared to control groups. 4) Furosemide treatment reduced the apoptotic index in transforming thick ascending limb of Henle's loop and collecting duct in renal medulla. 5) PCNA-positive cells in the transforming thick ascending limb of Henle's loop and the collecting duct in renal medulla were decreased in number in furosemide-treated groups compared to control groups. These finding suggest that renal hypoosmolality induced furosemide treatment decrease not only apoptosis but also cell proliferation and may retard renal papilla growth at least in neonatal rat kidney.
Animals ; Anion Exchange Protein 1, Erythrocyte ; Apoptosis ; Cell Proliferation ; Epithelium ; Extremities ; Furosemide* ; Humans ; Infant, Newborn ; Injections, Subcutaneous ; Kidney ; Morphogenesis* ; Proliferating Cell Nuclear Antigen ; Rats* ; Weights and Measures

OBJECTIVETo study the clinicopathological and immunohistochemical features of ectopic hamartomatous thymoma (EHT), and to discuss its histogenesis.

METHODSThe clinical and pathologic features of two EHT cases of were evaluated. Immunohistochemical study was performed by LSAB method using a panel of antibodies including AE1/AE3, CK5, CK7, CK8, CK20, EMA, vimentin, CD5, CD10, alpha-SMA, calponin, desmin, CD34, S-100 protein, CD57, GFAP, TTF-1 and CD99.

RESULTSBoth cases occurred in males aged 20 years and 40 years respectively. Each patient presented with a solitary mass, one located in the suprasternal fossa and the other in the left supraclavicular region for a period of 6 months and 2 months respectively. Grossly, the masses were well-circumscribed with spherical and ovoid appearance, measuring 5 cm and 3 cm in maximum diameter respectively. On cut section, they were gray-white in color and of soft consistency. Histologically, both tumors were composed of a mixture of spindle cells, epithelial cells and mature adipose tissue. The spindle cells element accounted 85% and 70% each in the two cases. They resembled fibroblasts in morphology and were arranged frequently in fascicular, woven or storiform patterns. Epithelial cells element represented nearly 10% in both cases. Most of the epithelial cells had a non-keratinization squamous appearance. They formed small solid islands and adamantinoma-like "nastomosing cords", or appeared as lining cells in large cystic spaces. In focal areas, glandular differentiation presented as small glands. A transition between the spindle cell and epithelium components could be also identified in some areas. Mature adipose tissue was irregularly distributed in the two tumors, about < 5% and 20% respectively. Immunohistochemically, the epithelial element expressed AE1/AE3, CK5, CK7, CK8 and EMA, whereas the spindle component expressed AE1/AE3, CK5, CK7, CK8, vimentin, CD10, CD34, alpha-SMA, MSA, and calponin. Both elements were negative for CK20, TTF-1, desmin, S-100 protein, CD57, GFAP and CD99.

CONCLUSIONSEHT is a benign tumor that occurs predominantly in the lower neck region of young to middle-aged males. Immunohistochemical study revealed myoepithelial differentiation of the spindle cells, suggesting EHT is a mixed tumor composed of epithelial and myoepithelial cells. EHT possibly originates from the remnants of cervical sinus of His, and therefore, may be renamed as branchial anlage mixed tumor.

Adult ; Anion Exchange Protein 1, Erythrocyte ; metabolism ; Choristoma ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Hamartoma ; Humans ; Keratins, Type II ; metabolism ; Lymphatic Diseases ; metabolism ; pathology ; surgery ; Male ; Mucin-1 ; metabolism ; Thymoma ; metabolism ; pathology ; surgery ; Thymus Neoplasms ; metabolism ; pathology ; surgery ; Vimentin ; metabolism