OBJECTIVETo observe abnormal metabolic changes caused by ischemic cerebral apoplexy and the regulating action of Tongsaimai pellets on abnormal metabolism by analyzing the change of small molecules in plasma of ischemic cerebral apoplexy rat. To find the potential biomarkers, and to explore metabolic mechanisms of Tongsaimai pellets.

METHODRat models of middle cerebral artery occlusion was established with electric coagulation, and rats were divided into 4 groups, model group, sham-operation group, Tongsaimai pellets group and positive control group. Tongsaimai pellets and positive control group were orally administrated by 13.2 g x kg(-1) x d(-1) of crude drugs and 32 mg x kg(-1) x d(-1) of Nimodipine respectively, m odel and sham-operation group by equal volume of distilled water for a week. Plasma of model and sham-operation group were collected, and plasma of Tongsaimai pellets and positive control group were collected on the 1st, 3rd , 7th day after administration. Endogenous metabolites of four groups were determined with GC-MS. Partial least squares discriminant analysis (PLS-DA) was applied to analyze multivariate data and set up model, and T-test was used in significant statistical analysis.

RESULTCompared with sham-operation group rats, pyruvic acid, taurine and hydroxyproline obviously increased in model group rats, while lactic acid, glyceric acid, aminomalonic acid, fructose, tryptophan and leucine significantly decreased, so these metabolites were potential metabolic biomarkers. These endogenous metabolites except taurine got restoration in Tongsaimai group rats.

CONCLUSIONAbnormal metabolite level in plasma can be certainly recovered by Tongsaimai pellets, and the treatment of Tongsaimai pellets can be connected with the regulation of related metabolic pathways.

Animals ; Brain Ischemia ; blood ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Fructose ; blood ; Glyceric Acids ; blood ; Hydroxyproline ; blood ; Lactic Acid ; blood ; Leucine ; blood ; Male ; Malonates ; blood ; Metabolomics ; methods ; Pyruvic Acid ; blood ; Rats ; Rats, Sprague-Dawley ; Stroke ; blood ; drug therapy ; Taurine ; blood ; Tryptophan ; blood

To compare the influence of the Bach Mai prepared AS-T preserving solution and Japanese Turumo firm prepared preserving solution in a same condition on the blood of 5 healthy persons of B group blood, who did not donate the blood. AS-T solution had stabilized and maintained the red blood cells, pH higher than 7 through a storage duration of 42 days. AS-T solution had maintained 2.3-DPG concentration in a level higher significantly in comparing with Terumo solution at the terminal points of the storage time PK and G6PD activities of the red blood cells mass preserved by AS-T solution as well as Terumo solution had decreased progressively in the duration of storage.
Erythrocytes ; 2,3-Diphosphoglycerate ; Pyruvate Kinase

BACKGROUND: The method utilizing high concentration glycerol is a common way to make frozen red bool cells (RBCs) for extended period, but it requires special deglycerolization equipment for washing after thawing. For economics reason, we could not have a cell washer for wahing the frozen RBCs, so attempted to use Haemonetics V50plus that we have had. METHODS: Twelve fresh packed RBCs were cryopreserved with 40% glycerol method. After 3 months, the RBCs were thawed and washed with Haemonetics V50plus. For the evaluation of the procedure, RBC reovery rate, osmolarity, 30 min saline stability (SS), % hemolysis, ATP, 2,3-DPG, LDH, potassium, and plasma hemoglobin were tested at 24hrs after washing. RESULTS: The RBC recovery rate was 82.1 +/- 4.5% (75.8-89.2) and two units of Frozen RBCs were under 80%. The Hb ATP and Hb 2,3-DPG of RBCs were 5.2 +/- 0.6 micro mol/g Hb (3.9-6.0) and 13.0 +/- 2.1 micro mol/g Hb (8.8-15.1). The supernatant osmalrity, potassium, plasma Hb and LDH were 352 +/- 7mosmol/kg H2O (342~367), 0.8 +/- 0.2 mmol/L (0.5~1.2), 0.8 +/- 0.3 mg/dL (0.4~1.1), 352 +/- 7 U/L (342~367). The 30min SS was 98.9 +/- 0.8. CONCLUSION: We could successfully freeze, thaw, and wash the frozen RBCs with Haemonetics V50 plus.
2,3-Diphosphoglycerate ; Adenosine Triphosphate ; Cryopreservation ; Erythrocytes* ; Glycerol ; Hemolysis ; Osmolar Concentration ; Plasma ; Potassium

High oxygen-affinity hemoglobin (Hb) variants and a 2,3-diphosphoglycerate (2,3-DPG) deficiency could cause congenital (familial) erythrocytosis. High oxygen-affinity Hb variants and a 2,3-DPG deficiency might result in low tissue oxygen tension left-shifted oxygen dissociation curves and reduction in the standard P₅₀ value (P(50,std), oxygen tension at which haemoglobin is 50% saturated). Hence, the P(50,std) value is considered while formulating diagnostic strategies for erythrocytosis. In this study, we established a reference range for P(50,std) using an International Federation of Clinical Chemistry and Laboratory Medicine-approved equation (Hill's equation) for individual single venous/arterial blood samples. Blood gas analysis results of 243 samples with oxygen saturation ranging from 40%–90% (Hb < 16 mg/dL) were selected. The reference range of P(50,std) was in the 2.5th–97.5th percentile, and was 25.9–27.3 mm Hg. Hill's equation is a simple approved method for evaluating the P(50,std) values. Only a single sample of venous or arterial blood and a blood gas analyser are required to obtain the P(50,std). Our study provides a useful tool for the diagnostic work-up of patients with erythrocytosis.
2,3-Diphosphoglycerate ; Blood Gas Analysis ; Chemistry, Clinical ; Humans ; Methods ; Oxygen ; Polycythemia* ; Reference Values

BACKGROUND: It is believed that an increase in the level of erythrocyte 2,3-diphosphoglycerate is an important compensatory mechanism that allows satisfactory tissue oxygenation in hypoxic patients. This study measured the level of 2,3-diphophoglycerate and the factors affecting its concentration, as well as the position of the oxygen-hemoglobin dissociation curve in patients with end-stage liver disease. METHODS: Fifty patients receiving liver transplantation (experimental group) and forty-five healthy donors (control group) were enrolled in this study. Arterial-mixed venous oxygen content difference, whole body oxygen delivery and consumption were measured after determining the hemodynamic parameters including cardiac output in the experimental group. The erythrocyte 2,3-diphosphoglycerate level was measured using assay procedures. The P50 value was used to determine the position of the oxygen-hemoglobin dissociation curve. RESULTS: The 2,3-diphosphoglycerate level was higher in patients with end-stage liver disease than in the controls. A 0.3 g/L increase in the 2,3-diphosphoglycerate concentration corresponded to a P50 increase in 1.2 mmHg with a rightward displacement of the oxygen-hemoglobin dissociation curve. The 2,3-diphosphoglycerate concentration showed a significant correlation with the PaO2, Ca-vO2, P50, and blood lactate level, but not with the hemodynamic parameters such as the cardiac index, oxygen delivery index, and pH. CONCLUSIONS: End-stage liver disease is associated with an increase in the level of the erythrocyte 2,3-diphosphoglycerate and P50. This is believed to be an important compensatory mechanism to allow better tissue oxygenation. An increase in the 2,3-diphosphoglycerate level correlates well with the oxygenation parameters, rather than with the hemodynamic parameters.
2,3-Diphosphoglycerate* ; Cardiac Output ; Erythrocytes* ; Hemodynamics ; Humans ; Hydrogen-Ion Concentration ; Lactic Acid ; Liver Diseases* ; Liver Transplantation ; Liver* ; Oxygen ; Tissue Donors

Fifteen healthy pregnant women at term, between the ages of 21 and 31 years, with their newborn babies, were investigated for their 2,3-DPG levels. All the babies were born by caesarean section. Seventeen nonpregnant women in the same age range as the mother group, without cardiopulmonary disease, metabolic disorder or severe anemia, were also investigated and used as the control group. Samples were collected from umbilical vessels in newborn babies and from arm vessels in mothers and nonpregnant women. The levels of 2,3-DPG were measured by the ultraviolet enzymatic method of Sigma Chemical Company. The results were as follows: 1. The 2,3-DPG lovels in teram of whole blood among the three groups were not significantly different. 2. The 2,3-DPG levels in terms of hemoglobin, the highest in the mother group and there was no difference the between the nonpregnant women and the newborn baby groups.
2,3-Diphosphoglycerate ; Anemia ; Arm ; Cesarean Section ; Female ; Humans ; Infant, Newborn* ; Metabolic Diseases ; Mothers* ; Pregnancy ; Pregnant Women ; Rabies*

BACKGROUND: Longer RBC storage will benefit blood banker by reduced outdating and patients by receiving fewer nonviable cells and RBC breakdown products as well as by having the opportunity to participate in markedly improved autologous storage system. METHODS: We rejuvenated whole blood 10 units and RBC 15 units, on 1-3 days after expiration, then they were divided into four, washed, and suspended with four kinds of solutions containing saline adenine glucose, mannitol, and phosphate, in closed system. For the evaluation of RBCs, 30 min saline stability (SS), 2 h SS, % hemolysis, ATP, 2,3-DPG, P50, LDH, potassium, and plasma hemoglobin were tested at every week during 42 days. RESULTS: On 35th day, 30 min SS of RBCs was higher than 88% and % hemolysis of RBCs was lower than 1% in solutions containing phosphate. ATP of RBCs was the 50% of the reference value of healthy persons and 2,3-DPG of RBCs was relatively higher in solutions containing phosphate (SAGMP1, 2, 3) than in solution without phosphate (SAGM). CONCLUSION: We could successfully rejuvenate outdated RBCs and extend the expiration date to additive 35 days, but for practical use, post transfusion RBC survival and safety should be evaluated in the future.
2,3-Diphosphoglycerate ; Adenine ; Adenosine Triphosphate ; Blood Banks ; Erythrocytes* ; Glucose ; Hemolysis ; Humans ; Mannitol ; Plasma ; Potassium ; Reference Values ; Rejuvenation

OBJECTIVE: To explore the pathogenesis of erythrocytosis by detecting the key enzymes of glucose metabolism and glucose transporter in bone marrow erythrocytes of chronic mountain sickness (CMS), and analyzing its correlation with hemoglobin. METHODS: Twenty CMS patients hospitalized in Qinghai Provincial People's Hospital from January 2019 to December 2020 were selected as CMS group. Twenty males with leukocyte count > 3.5×10⁹/L who had accepted bone marrow aspiration and had normal result were taken as control group. The mRNA and protein expression of key enzymes and glucose transporter in glucose metabolism in bone marrow CD71⁺ erythrocytes were detected by real time qPCR and Western blot, respectively. Glucose, lactic acid and 2,3-diphosphoglycerate in the bone marrow supernatant and serum were tested by ELISA. The mRNA and protein expression of key enzymes and glucose transporter, glucose, lactic acid and 2,3-diphosphoglycerate of the two groups were compared. Pearson correlation was used to analyze the correlation between key enzymes, glucose transporter in glucose metabolism in bone marrow CD71⁺ erythrocytes and hemoglobin. RESULTS: The expression of HK2, GLUT1 and GLUT2 mRNA in the CMS group were higher than those in the control group (P<0.001), while the expression of HK1, OGDH and COX5B mRNA were not different. The expression of HK2, GLUT1 and GLUT2 protein in the CMS group were higher than those in the control group (P<0.05). The levels of glucose and lactic acid in the bone marrow supernatant and serum in the CMS group were not different from those in the control group, while the level of 2,3-diphosphoglycerate was higher (P<0.001). Both HK2 and GLUT2 proteins were positively correlated with hemoglobin (r=0.511, 0.717). CONCLUSION CMS patients may increase glycolysis by increasing the expression of HK2, and promote the utilization of glucose through high expression of GLUT1 and GLUT2 to meet the need of energy supply.
Male ; Humans ; Altitude Sickness/metabolism* ; Glucose Transporter Type 1 ; 2,3-Diphosphoglycerate ; Hemoglobins ; Chronic Disease ; RNA, Messenger ; Phenotype ; Glucose

OBJECTIVE: To investigate the anti-oxidative effect of ethyl pyruvate (EP) and taurine (TAU) on the quality of red blood cells stored at 4±2 ℃, hemolysis, energy metabolism and lipid peroxidation of the red blood cells in the preservation solution were studied at different intervals. METHODS: At 4±2 ℃, the deleukocyte red blood cells were stored in the citrate-phosphate-dextrosesaline-adenine-1 (CPDA-1) preservation (control group), preservation solution with EP (EP-AS), and TAU (TAU-AS) for long-term preservation. The enzyme-linked immunoassay and automatic blood cell analyzer were used to detect hemolysis and erythrocyte parameters. Adenine nucleoside triphosphate (ATP), glycerol 2,3-diphosphate (2,3-DPG) and malondialdehyde (MDA) kits were used to test the ATP, 2,3-DPG and MDA concentration. RESULTS: During the preservation, the rate of red blood cell hemolysis in EP-AS and TAU-AS groups were significantly lower than that in CPDA-1 group (P<0.01). The MCV of EP-AS group was increased with the preservation time (r=0.71), while the MCV of the TAU-AS group was significantly lower than that in the other two groups (P<0.05). The concentration of ATP and MDA in EP-AS and TAU-AS groups were significantly higher than that in CPDA-1 group at the 14th day (P<0.01). The concentrations of 2,3-DPG in the EP-AS and TAU-AS groups were significantly higher than that in the CPDA-1 group from the 7th day (P<0.01). CONCLUSION EP and TAU can significantly reduce the red blood cell hemolysis rate, inhibit the lipid peroxidation level of red blood cells, and improve the energy metabolism of red blood cells during storage. The mechanism of EP and TAU may be related to their antioxidation and membrane protection effect, so as to improve the red blood cell quality and extend the preservation time.
2,3-Diphosphoglycerate/metabolism* ; Adenine ; Adenosine Triphosphate/metabolism* ; Blood Preservation ; Citrates/pharmacology* ; Erythrocytes/metabolism* ; Glucose/pharmacology* ; Hemolysis ; Humans ; Pyruvates ; Taurine/pharmacology*

BACKGROUND: The use of autologous transfusion is gradually increasing since it eliminates transfusion-transmitted viral diseases, and avoids the risk of alloimmunization of red blood cells and posttransfusion graft-versus-host disease. The majority of premature neonates born at less than 1500 g need one or more red blood cell transfusion during the hospitalization and cord blood is considered as the most ideal blood for neonate autologous transfusion. In order to evaluate the adequacy of stored cord blood for autologous transfusion for neonates, the levels of plasma free hemoglobin, red blood cell adenosine triphosphate (ATP) and 2,3-diphosphoglycerate (2,3-DPG) were measured at the time of collection, and then a week interval by 4 weeks. METHODS: The cord blood was collected in a single donor bag with CPDA-1 by aseptic technique from 28 newborns, stored for 28 days at 4degrees C, and changes in the levels of plasma free hemoglobin, red blood cell ATP and 2,3-DPG were measured at the time of collection, and then a week interval by 4 weeks for 26 cord bloods which were not presented with any bacterial growth during the storage. RESULTS: At the time of sampling, hemolysis was 0.11+/-0.16%, and intracellular ATP and 2,3-DPG were 3.74+/-0.99 mumol/g Hb and 11.67+/-1.21 mumol/g Hb, respectively. During the storage, hemolysis gradually increased to 0.61+/-1.09% on 28 days (p<0.05). ATP gradually decreased to 2.98+/-0.92 mumol/g Hb (80% of initial level) on 28 days(p<0.05). The levels of 2,3-DPG were 4.20+/-0.87 mumol/g Hb (about 35% of initial level) on 7 days(p<0.05) and 1.16+/-0.74 mumol/g Hb (less than 10% of initial level) on 28 days (p<0.001). CONCLUSIONS: In conclusion, ATP and 2,3-DPG levels of cord blood that are related to the viability of red blood cells during the storage were similar to those of adults. Thus the cord blood appeared to be an appropriate source for neonate autologous transfusion, however, more intensive studies on the effects of 2,3-DPG and metabolic products in vivo are necessary since physical conditions and physiology of the red blood cells in the neonates are different in many aspects from those of adults and children.
2,3-Diphosphoglycerate* ; Adenosine Triphosphate* ; Adenosine* ; Adult ; Child ; Erythrocyte Transfusion ; Erythrocytes ; Fetal Blood* ; Graft vs Host Disease ; Hemolysis ; Hospitalization ; Humans ; Infant, Newborn ; Physiology ; Plasma* ; Tissue Donors ; Virus Diseases