Our study aims to explore the effects of lentivirus-mediated microRNA-124 (miR-124) gene-modified bone marrow mesenchymal stem cell (BMSC) transplantation on the repair of spinal cord injury (SCI) in rats. BMSCs were isolated from the bone marrow of rats. The target gene miR-124 was identified using a luciferase-reporter gene assay. Seventy-two rats were selected for construction of the SCI model, and the rats were randomly divided into the blank group, sham group, SCI group, negative control (NC) group, overexpressed miR-124 group and si-PDXK group. The mRNA expression of miR-124 and the mRNA and protein expression of pyridoxal kinase (PDXK) were detected by quantitative real-time polymerase chain reaction and western blotting. The locomotor capacity of the rats was evaluated using the Basso, Beattie and Bresnahan (BBB) scale. Brdu, neuron-specific enolase (NSE), neurofilament (NF) and microtubule-associated protein 2 (MAP2) were detected using immunohistochemistry. The expression levels of thyrotropin-releasing hormone (TRH), prostacyclin (PGI2) and gangliosides (GM) were measured using an enzyme-linked immunosorbent assay. PDXK was identified as the target gene of miR-124. The overexpressed miR-124 group exhibited higher miR-124 expression than the SCI, NC and si-PDXK groups. Compared with the SCI and NC groups, the PDXK expression was downregulated in the overexpressed miR-124 and si-PDXK groups, and the BBB scores were significantly increased 7, 21 and 35 days after transplantation. The double-labeled positive cell densities (Brdu+NSE/NF/MAP2) and the expression levels of TRH, PGI2 and GM in the overexpressed miR-124 group were significantly higher than those in the NC and SCI groups. These results indicated that miR-124 targeted PDXK to accelerate the differentiation of BMSCs into neurocytes and promote SCI repair.
Animals ; Blotting, Western ; Bone Marrow* ; Bromodeoxyuridine ; Cell Count ; Enzyme-Linked Immunosorbent Assay ; Epoprostenol ; Gangliosides ; Immunohistochemistry ; Intermediate Filaments ; Mesenchymal Stem Cell Transplantation* ; Mesenchymal Stromal Cells* ; Microtubule-Associated Proteins ; Phosphopyruvate Hydratase ; Pyridoxal Kinase ; Rats* ; Real-Time Polymerase Chain Reaction ; RNA, Messenger ; Spinal Cord Injuries* ; Spinal Cord* ; Thyrotropin-Releasing Hormone

BACKGROUND: Pyridoxal-5'-phosphate (P5P), a coenzyme of the aspartate aminotransferase (AST) and alanine aminotransferase (ALT) reactions, is required to measure aminotransferase levels (IFCC method). However, a modified IFCC method that uses a reagent devoid of P5P is commonly used in laboratories in Korea. To determine the differences between the two methods, we compared aminotransferase levels measured by using the IFCC method and modified IFCC method. METHODS: Serum levels of AST and ALT, with and without P5P, were measured in 2,318 patients. Based on the allowable limits of performance set by the Royal College of Pathologists of Australasia (RCPA), differences between the two methods were analyzed under various conditions. RESULTS: Higher AST and ALT values were obtained by the IFCC method compared to modified IFCC method, showing significant differences between the two methods (AST, 5.8±14.2 IU/L; ALT, 2.8±6.9 IU/L) (P<0.001). Values exceeding RCPA criteria were more frequently observed in emergency orders (AST, 65.8%; ALT, 14.4%) than in routine orders (AST, 3.2%; ALT, 9.6%), as well as in inpatient wards (AST, 70.4%; ALT, 18.5%) compared to outpatient clinics (AST, 56.6%; ALT, 10.0%). However, the differences between the two methods were not significant among the disease groups, except for the acute myocardial infarction group. CONCLUSIONS: The method using reagents without P5P underestimated aminotransferase activity. The effect of P5P was more significant in patients with acute myocardial infarction, considered as P5P-deficient. In conclusion, the IFCC method with P5P should be applied for measuring AST and ALT serum levels.
Alanine Transaminase ; Ambulatory Care Facilities ; Aspartate Aminotransferases ; Australasia ; Emergencies ; Humans ; Indicators and Reagents ; Inpatients ; Korea ; Liver Function Tests ; Methods ; Myocardial Infarction ; Pyridoxal Phosphate

BACKGROUND/OBJECTIVES: Recent research regarding vitamin B6 status including biochemical index is limited. Thus, this study estimated intakes and major food sources of vitamin B6; determined plasma pyridoxal 5'-phosphate (PLP); and assessed vitamin B6 status of Korean adults. MATERIALS/METHODS: Three consecutive 24-h diet recalls and fasting blood samples were collected from healthy 20- to 64-year-old adults (n = 254) living in the Seoul metropolitan area, cities of Kwangju and Gumi, Korea. Vitamin B6 intake and plasma PLP were analyzed by gender and by vitamin B6 supplementation. Pearson's correlation coefficient was used to determine associations of vitamin B6 intake and plasma PLP. RESULTS: The mean dietary and total (dietary plus supplemental) vitamin B6 intake was 1.94 +/- 0.64 and 2.41 +/- 1.45 mg/day, respectively. Median (50th percentile) dietary intake of men and women was 2.062 and 1.706 mg/day. Foods from plant sources provided 70.61% of dietary vitamin B6 intake. Only 6.3% of subjects consumed total vitamin B6 less than Estimated Average Requirements. Plasma PLP concentration of all subjects was 40.03 +/- 23.71 nmol/L. The concentration of users of vitamin B6 supplements was significantly higher than that of nonusers (P < 0.001). Approximately 16% of Korean adults had PLP levels < 20 nmol/L, indicating a biochemical deficiency of vitamin B6, while 19.7% had marginal vitamin B6 status. Plasma PLP concentration showed positive correlation with total vitamin B6 intake (r = 0.40984, P < 0.0001). CONCLUSIONS: In this study, vitamin B6 intake of Korean adults was generally adequate. However, one-third of subjects had vitamin B6 deficiency or marginal status. Therefore, in some adults in Korea, consumption of vitamin B6-rich food sources should be encouraged.
Adult ; Diet ; Fasting ; Female ; Gwangju ; Gyeongsangbuk-do ; Humans ; Korea ; Male ; Middle Aged* ; Morinda ; Plants ; Plasma ; Pyridoxal ; Seoul ; Vitamin B 6 Deficiency ; Vitamin B 6* ; Vitamins

Interstitial cells of Cajal (ICCs) are the pacemaker cells in the gastrointestinal tract, and histamine is known to regulate neuronal activity, control vascular tone, alter endothelial permeability, and modulate gastric acid secretion. However, the action mechanisms of histamine in mouse small intestinal ICCs have not been previously investigated, and thus, in the present study, we investigated the effects of histamine on mouse small intestinal ICCs, and sought to identify the receptors involved. Enzymatic digestions were used to dissociate ICCs from small intestines, and the whole-cell patch-clamp configuration was used to record potentials (in current clamp mode) from cultured ICCs. Histamine was found to depolarize resting membrane potentials concentration dependently, and whereas 2-PEA (a selective H1 receptor agonist) induced membrane depolarizations, Dimaprit (a selective H2-agonist), R-alpha-methylhistamine (R-alpha-MeHa; a selective H3-agonist), and 4-methylhistamine (4-MH; a selective H4-agonist) did not. Pretreatment with Ca(2+)-free solution or thapsigargin (a Ca(2+)-ATPase inhibitor in endoplasmic reticulum) abolished the generation of pacemaker potentials and suppressed histamine-induced membrane depolarization. Furthermore, treatments with U-73122 (a phospholipase C inhibitor) or 5-fluoro-2-indolyl des-chlorohalopemide (FIPI; a phospholipase D inhibitor) blocked histamine-induced membrane depolarizations in ICCs. On the other hand, KT5720 (a protein kinase A inhibitor) did not block histamine-induced membrane depolarization. These results suggest that histamine modulates pacemaker potentials through H1 receptor-mediated pathways via external Ca2+ influx and Ca2+ release from internal stores in a PLC and PLD dependent manner.
Animals ; Carbazoles ; Cyclic AMP-Dependent Protein Kinases ; Dimaprit ; Domperidone ; Estrenes ; Gastric Acid ; Gastrointestinal Tract ; Hand ; Histamine ; Indoles ; Interstitial Cells of Cajal ; Intestine, Small ; Membrane Potentials ; Membranes ; Methylhistamines ; Mice ; Neurons ; Permeability ; Phospholipase D ; Pyridoxal ; Pyrroles ; Pyrrolidinones ; Thapsigargin ; Type C Phospholipases

PURPOSE: The authors of the present study investigated whether pyridoxal 5'-phosphate (PLP), an active coenzyme of vitamin B6, could inhibit the development of diabetic retinopathy in streptozotocin (STZ)-induced diabetic rats. METHODS: Seven-week-old Spraque-Dawley rats (n = 20) were used in the present study. STZ (70 mg/kg) was injected intraperitoneally to induce diabetes. Blood glucose and body weight were monitored. Intraperitoneal injections of 5 microg and 50 microg PLP were administered every two days from the second week of induced diabetes. During the third week of PLP injections, the concentration level of plasma homocysteine was measured. In addition, functional status was examined by vitreous fluorophotometer and anatomical status by vascular endothelial growth factor (VEGF) staining in the retina. RESULTS: Based on vitreous fluorophotometry examination, the PLP injection group proved to have a lower level of fluorescein concentration in the vitreous. Additionally, immunohistochemical staining revealed down-regulation of VEGF expression in the PLP group. In addition, the PLP group had a lower plasma homocysteine concentration. However, an over-dosage injection of PLP did not appear to have any noticeable impact on the treatment of diabetes. CONCLUSIONS: PLP, an active coenzyme of vitamin B6, proved to have inhibitory effects on VEGF expression and vascular leakage in the diabetic rat retina.
Animals ; Blood Glucose ; Body Weight ; Diabetic Retinopathy ; Down-Regulation ; Fluorescein ; Fluorophotometry ; Homocysteine ; Injections, Intraperitoneal ; Plasma ; Pyridoxal ; Rats ; Retina ; Streptozocin ; Vascular Endothelial Growth Factor A ; Vitamin B 6

The purpose of this study was to assess vitamin B6 intake and status in Korean patients with newly diagnosed type 2 diabetes. Sixty-four patients with newly diagnosed type 2 diabetes and 8-11% glycated hemoglobin (A1C), along with 28 age-matched non-diabetic subjects, participated. Dietary vitamin B6 intake was estimated by the 24 hour recall method and plasma pyridoxal 5'-phosphate (PLP) was measured. There was a significant difference in daily total calorie intake between the diabetic and non-diabetic groups (1,917 +/- 376 vs 2,093 +/- 311 kcal). There were no differences in intake of total vitamin B6 (2.51 +/- 0.91 vs 2.53 +/- 0.81 mg/d) or vitamin B6/1,000 kcal (1.31 +/- 0.42 vs 1.20 +/- 0.32 mg) between the diabetic and non-diabetic groups, andI intakes of total vitamin B6 were above the Korean RDA in both groups (180.0 +/- 57.9 vs 179.0 +/- 65.4). There was a higher percentage of diabetic subjects whose plasma PLP concentration was < 30 nmol/L compared to non-diabetic group. Plasma PLP levels tended to be lower in the diabetic subjects than in the non-diabetic subjects, although the difference was not statistically significant due to a large standard deviation (80.0 +/- 61.2 nmol/L vs 68.2 +/- 38.5 nmol/L). Nevertheless, plasma PLP levels should be monitored in pre-diabetic patients with diabetic risk factors as well as in newly diagnosed diabetic patients for long-term management of diabetes, even though this factor is not a major risk factor that contributes to the development of degenerative complications in certain patients.
Fibrinogen ; Hemoglobins ; Humans ; Nutrition Policy ; Plasma ; Pyridoxal ; Risk Factors ; Vitamin B 6 ; Vitamins

The ginkgo nuts are commonly eaten because of its nutritive value. It also taken as an herbal medicine for its antitussive and expectorant properties throughout Korea, Japan, and China. A range of symptoms of ginkgo nut intoxication including vomiting, abdominal pain, irritability, headaches, and seizures have been reported. It can be fetal in severe cases if emergency care was not available. It was discovered that the toxicity of ginkgo nuts was due to 4-O-methoxypyridoxine. The convulsion can be prevented by administration of anticonvulsants and pyridoxal phosphate. We report a 6-year-old male, without any past or family histories of epilepsy, who presented with generalized tonic clonic seizures. About 7 hours before the seizure, he had consumed approximately 40 roasted ginkgo nuts. He exhibited vomiting and abdominal pain 5 hours after ingestion. After treatment with diazepam and pyridoxal phosphate, his symptoms were resolved and electroencephalogram was normalized. We report on the patient and emphasize pediatricians and parents who have children should be recognizing that neurotoxicity of ginkgo nuts, particularity convulsion inducing effect, can occur occasionally.
Abdominal Pain ; Anticonvulsants ; Child ; China ; Diazepam ; Eating ; Electroencephalography ; Emergency Medical Services ; Epilepsy ; Ginkgo biloba ; Headache ; Herbal Medicine ; Humans ; Japan ; Korea ; Male ; Nutritive Value ; Nuts ; Parents ; Pyridoxal Phosphate ; Pyridoxine ; Seizures ; Vomiting

This study was conducted to investigate the effects of extremely low frequency electromagnetic fields (EMF) on signal pathway in plasma membrane of cultured cells (RAW 264.7 cells and RBL 2H3 cells), by measuring the activity of phospholipase A2 (PLA2), phospholipase C (PLC) and phospholipase D (PLD). The cells were exposed to the EMF (60 Hz, 0.1 or 1 mT) for 4 or 16 h. The basal and 0.5 microM melittin-induced arachidonic acid release was not affected by EMF in both cells. In cell-free PLA2 assay, we failed to observe the change of cPLA2 and sPLA2 activity. Also both PLC and PLD activities did not show any change in the two cell lines exposed to EMF. This study suggests that the exposure condition of EMF (60 Hz, 0.1 or 1 mT) which is 2.4 fold higher than the limit of occupational exposure does not induce phospholipases-associated signal pathway in RAW 264.7 cells and RBL 2H3 cells.
Arachidonic Acid ; Cell Line ; Cell Membrane ; Cells, Cultured ; Electromagnetic Fields ; Magnets ; Occupational Exposure ; Phospholipase D ; Phospholipases ; Phospholipases A2 ; Pyridoxal ; Signal Transduction ; Type C Phospholipases

The purpose of this study was to establish the selection of indicators for estimating and factors affecting the requirement of vitamin B6. There has been a need to establish the human requirements of vitamin B6 since vitamin B6 is thought to be involved in more than one hundred biochemical reactions as a coenzyme in the metabolism of amino acids, glucose, and lipid, and the synthesis of neurotransmitters. For the review of the literature, this study included from early findings of the sixties to studies of 2009. This study suggests that plasma pyridoxal 5' phosphate (PLP) is the best single indicator of vitamin B6 status for the healthy but not for the non-healthy. Erythrocyte aspartate aminotransferase and alanine aminotransferase activation by PLP as an indirect measure and urinary 4-pyridoxic acid excretion as a direct measure are useful as supporting indicators. Bioavailability, nutrient interaction, physiological need, and chronic diseases may increase the requirement for vitamin B6. However, these effects can not be quantified due to insufficient evidences.
Alanine Transaminase ; Amino Acids ; Aspartate Aminotransferases ; Biological Availability ; Chronic Disease ; Erythrocytes ; Glucose ; Humans ; Neurotransmitter Agents ; Plasma ; Pyridoxal ; Pyridoxic Acid ; Vitamin B 6 ; Vitamins

Purified hemoglobin was modified with pyridoxal 5-phosphate(PLP) and polymerized with glutaric dialdehyde(GDA) to get the products. By comparison of the physical, chemical and biological properties of different procedures for modification before and after polymerization, there is no significant difference in molecular distribution, methemoglobin(MetHb) concentration, oxygen carrier capacity, P50 and spectra. Furthermore, the procedure of modification after polymerization can save PLP greatly and decrease cost greatly. So the procedure of modification after polymerization is a better way in research and production. The addition of GDA could control the increasing of MetHb. By comparison on the physical, chemical and biological properties of different procedures, there is no significant difference in molecular distribution, MetHb concentration, oxygen carrier capacity and spectra between the procedure of adding GDA before PLP and that after PLP. But the P50 of adding GDA before PLP is much lower than that after PLP. So the procedure of adding GDA after PLP is a better way.
Blood Substitutes ; Chemical Phenomena ; Glutaral ; chemistry ; Hemoglobins ; chemistry ; Humans ; Methemoglobin ; chemistry ; Pyridoxal Phosphate ; analogs & derivatives ; chemistry