1.Characterization of Streptococcus oligofermentans sucrose metabolism demonstrates reduced pyruvic and lactic acid production.
Xu-Dong BAO ; Lin YUE ; Xue-Jun GAO
Chinese Medical Journal 2011;124(21):3499-3503
BACKGROUNDStreptococcus (S.) oligofermentans is a newly identified bacteria with a yet to be defined mechanism of sucrose metabolism that results in acid production. This study aimed to investigate the biochemical mechanisms of S. oligoferm-entans glucose metaolism.
METHODSThe S. oligofermentans LMG21532, Lactobacillus (L.) fermentum 38 and the S. mutans UA140 were used to characterize sucrose metabolism by measuring lactate dehydrogenase (LDH) activity and lactic acid production. Continuous dynamics and high performance capillary electrophoresis were used to determine LDH activity and lactic acid production, respectively, from bacteria collected at 0, 10 and 30 minutes after cultured in 10% sucrose.
RESULTSThese analyses demonstrated that LDH activity of the three bacterial strains examined remained stable but significantly different throughout the sucrose fermentation process. The S. oligofermentans LDH activity ((0.61 ± 0.05) U/mg) was significantly lower than that of L. fermentum ((52.91 ± 8.97) U/mg). In addition, the S. oligofermentans total lactate production ((0.048 ± 0.021) mmol/L) was also significantly lower than that of L. fermentum ((0.958 ± 0.201) mmol/L). Although the S. oligofermentans LDH production was almost double of that produced by S. mutans ((0.32 ± 0.07) U/mg), lactic acid production was approximately one sixth that of S. mutans ((0.296 ± 0.058) mmol/L). Additional tests examining pyruvic acid production (the LDH substrate) demonstrated that lactic acid concentrations correlated with pyruvic acid production. That is, pyruvic acid production by S. oligofermentans was undetectable following sucrose incubation, however, (0.074 ± 0.024) and (0.175 ± 0.098) mmol/L pyruvic acid were produced by S. mutans and L. fermentum, respectively.
CONCLUSIONS. oligofermentans is incapable of fermenting carbohydrates to produce enough pyruvic acid, which results in reduced lactic acid production.
L-Lactate Dehydrogenase ; metabolism ; Lactic Acid ; metabolism ; Pyruvic Acid ; metabolism ; Streptococcus ; metabolism ; Sucrose ; metabolism
2.Activity and isoenzyme spectra of lactate dehydrogenase of plateau zokor (Myospalax baileyi) in different season.
Deng-Bang WEI ; Hong-Yan YU ; Jian-Mei ZHANG ; Xiao-Jun WANG ; Lian WEI
Chinese Journal of Applied Physiology 2007;23(3):365-369
AIMTo explore the adapting metabolic mechanisms of the plateau zokors to the hypoxic-hypercapnic environment.
METHODSThe activities of lactate dehydrogenase in serum and tissues, and the content of lactate in serum of plateau zokors in spring, summer and autumn were determined by using method of enzyme analysis. The spectrums of lactate dehydrogenase isoenzymes in serum and tissues of plateau zokors in spring, summer and autumn were analyzed by using method of the discontinuous systemic poly-acrylamide perpendicular plank gel electrophoresis.
RESULTSThe activities of lactate dehydrogenase in serum had obvious seasonally difference that were higher in spring and lower in autumn, and the content of lactate in serum showed same changing pattern. The spectrums of lactate dehydrogenase isoenzymes in serum showed five bands that were LDH1, LDH2, LDH3, LDH4 and LDH5 from positive pole to negative pole respectively, it showed clearly two bands in serum of summer that were LDH4 and LDH5 and one band in serum of autumn that was LDH5. The activities of LDH in tissues of skeleton muscle, cardiac muscle and brain were higher compared with the other tissues, it decreased markedly from spring to summer to autumn. In tissues of liver, kidney and lungs, activities of LDH were lower. Activities of LDH in livers, were significantly higher in spring compared that in summer and autumn, which had no obvious difference between summer and autumn. Activities of LDH in kidneys and lungs, showed no obviously difference between spring and summer, which decreased markedly in autumn. The spectrums of lactate dehydrogenase isoenzymes in tissues of cardiac muscle, liver, lungs, kidney, brain and skeleton muscle showed five bands, the spectrums were obvious different in different tissues, and the content of LDH isoenzymes showed seasonal changes in different tissues.
CONCLUSIONGlycolysis levels in plateau zokors had obvious seasonally change which increased in spring and decreased in autumn significantly. It related to the activity of plateau zokors in different seasons and seasonal fluctuation of oxygen and carbon dioxide in burrows of plateau zokors.
Animals ; Carbon Dioxide ; metabolism ; Isoenzymes ; analysis ; metabolism ; L-Lactate Dehydrogenase ; analysis ; metabolism ; Rodentia ; metabolism ; Seasons
3.Study on the content of myoglobin and the activity of lactate dehydrogenase and malate dehydrogenase in skeletal muscle of tibetan antelope.
Lan MA ; Ying-Zhong YANG ; Ri-Li GE
Chinese Journal of Applied Physiology 2012;28(2):118-121
OBJECTIVETo explore the adaptive mechanism to hypoxia in skeletal muscle of tibetan antelope.
METHODSTibetan sheep which living at the same altitude (4 300 m) with tibetan antelope and low altitude (1 800 m) sheep as control, the content of myoglobin (Mb) and lactic acid (LA), the activity of lactate dehydrogenase (LDH) and malate dehydrogenase (MDH) in skeletal muscles among three animals were analyzed by spectrophotometer.
RESULTSThe content of myoglobin in skeletal muscle of tibetan antelope significantly higher than that of tibetan sheep and low altitude sheep (P < 0.05). And the content of LA in skeletal muscle of tibetan antelope significantly lower than that of tibetan sheep and low altitude sheep (P < 0.05), activity of LDH and MDH in skeletal muscle was significantly lower and higher respectively than that of tibetan sheep and low altitude sheep (P < 0.05). There was no significant difference between tibetan sheep and low altitude sheep.
CONCLUSIONTibetan antelope may improve their ability to get oxygen under hypoxia by increasing the content of myoglobin in skeletal muscle, and the proportion of aerobic metabolism is high in skeletal muscle, it may be relate that with high myoglobin content in skeletal muscle, we suppose that high myoglobin content in skeletal muscle of tibetan antelope might be one of the molecular basis to adapt hypoxia.
Altitude ; Animals ; Antelopes ; metabolism ; physiology ; Hypoxia ; metabolism ; L-Lactate Dehydrogenase ; metabolism ; Malate Dehydrogenase ; metabolism ; Muscle, Skeletal ; metabolism ; Myoglobin ; metabolism
4.Change in gastrocnemius dystrophin and metabolic enzymes and increase in high-speed exhaustive time induced by hypoxic training in rats.
Yu-Ming XU ; Jun-Ping LI ; Rui-Yuan WANG
Acta Physiologica Sinica 2012;64(4):455-462
The aim of the present study was to explore the changes and roles of dystrophin and membrane permeability in hypoxic training. Seventy-two 8-week-old Sprague Dawley (SD) rats were randomly divided into 4 groups, normoxic non-train (NC), normoxic train (NT), hypoxic non-train (HC), and hypoxic train (HT) groups. The rats of each group were randomly divided into three subgroups, non-exhaustive, low-speed exhaustive test and high-speed exhaustive test subgroups. Rats in hypoxia groups lived and were trained in a condition of 12.7% oxygen concentration (equal to the 4 300 m altitude). NT and HT groups received 4 weeks of training exercise. Then the rats in all non-exhaustive subgroups were sacrificed, and gastrocnemii were sampled for the measurements of lactate dehydrogenase (LDH), succinatedehydrogenase (SDH), malate dehydrogenase (MDH) activities. Moreover, serum LDH activity was analyzed. Low-speed exhaustive test and high-speed exhaustive test subgroups received exhaustive tests with 20 (71% VO2max) and 30 m/min speed (86% VO2max), respectively, and their exhaustive times were recorded. The results showed that, compared with normoxic groups, the weights in hypoxia groups exhibited slower increase. The level of dystrophin in HT group without exhaustion test didn't change significantly. The muscle MDH activities were markedly affected by the different oxygen concentration, training and their interaction (P < 0.05), whereas the muscle LDH activities were only affected by the different oxygen concentration (P < 0.05). Serum LDH activities were affected by the interaction of the different oxygen concentration and training (P < 0.05), showing decreased muscle LDH and increased blood LDH activities. The exhaustion time were markedly affected by the different test speed, training and their interaction (P < 0.05), and also affected by the interaction of the different oxygen concentration and training (P < 0.05), but didn't affected by oxygen concentration. The exhaustive time of HT high-speed exhaustive test subgroup was more than NT high-speed exhaustive test subgroup in 30 m/min exhaustion test. Compared with NT high-speed exhaustive test subgroup, HT high-speed exhaustive test subgroup had an earlier fatigue in the test, but had a rapid recovery. These results suggested that hypoxic training can effectively increase the rats' high-speed exhaustive time. The mechanism may be related to an increase in serum LDH caused by the increased membrane permeability after hypoxic training.
Altitude
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Animals
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Dystrophin
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metabolism
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Fatigue
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Hypoxia
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L-Lactate Dehydrogenase
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metabolism
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Malate Dehydrogenase
;
metabolism
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Muscle, Skeletal
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enzymology
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Physical Conditioning, Animal
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Rats
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Rats, Sprague-Dawley
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Succinate Dehydrogenase
;
metabolism
5.Hypoxic adaptation of the hearts of plateau zokor (Myospalax baileyi) and plateau pika (Ochotona curzoniae).
Xin-Zhang QI ; Xiao-Jun WANG ; Shi-Hai ZHU ; Xin-Feng RAO ; Lian WEI ; Deng-Bang WEI
Acta Physiologica Sinica 2008;60(3):348-354
Plateau zokor (Myospalax baileyi) and plateau pika (Ochotona curzniae) are native to the Qinghai-Tibet plateau. To study their adaptive mechanisms, the ratios of heart weight to body weight (HW/BW) and right to left ventricular plus septum weights [RV/(LV+S)] were determined; the microvessel density (MVD) of cardiac muscle were measured by immunohistochemical staining; the numerical density on area (N(A)), volume density (V(V)), specific surface (δ), and surface density (S(V)) of mitochondria were obtained by microscopy and stereology; the contents of myoglobin (Mb) and lactic acid (LD), and the activity of lactate dehydrogenase (LDH) in cardiac muscle were analyzed by spectrophotometer. The results showed that the HW/BW of plateau zokor [(4.55±0.26)%] and plateau pika [(4.41±0.38)%] was significantly greater than that of Sprague-Dawley (SD) rat [(3.44±0.41)%] (P<0.05), but the RV/(LV+S) [(22.04±1.98)%, (25.53±3.41)%] was smaller than that of SD rats [(44.23±3.87)%] (P<0.05). The MVD and N(A) of cardiac muscle were 1688.631±250.253 and 0.768±0.123 in SD rat, 2002.888±367.466 and 0.868±0.159 in plateau pika and 2 990.643±389.888 and 1.012±0.133 in plateau zokor. The V(V) of mitochondria in plateau zokor (0.272±0.045) was significantly lower than that in plateau pika (0.343±0.039) and SD rat (0.321±0.048) (P<0.05), while the δ of mitochondria in plateau zokor (9.409±1.238) was higher than that in plateau pika (6.772±0.892) and SD rat (7.287±1.373) (P<0.05). The S(V) of mitochondria in plateau pika (2.322±0.347) was not obviously different from that in plateau zokor (2.468±0.380) and SD rat (2.227±0.377), but that in plateau zokor was significantly higher than that in SD rat (P<0.05). The contents of Mb in cardiac muscle of plateau zokor [(763.33±88.73) nmol/g] and plateau pika [(765.96±28.47) nmol/g] were significantly higher than that of SD rat [(492.38±72.14) nmol/g] (P<0.05), the content of LD in plateau zokor [(0.57±0.06) mmol/L] was obviously higher than that in plateau pika [(0.45±0.06) mmol/L] and SD rat [(0.48±0.02) mmol/L] (P<0.05), and the activity of LDH in plateau zokor [(16.90±2.00) U/mL] and plateau pika [(20.55±2.46) U/mL] were significantly lower than that in SD rat [(38.26±6.78) U/mL] (P<0.05). The percentage of LDH-H in cardiac muscle decreased in order in plateau zokor, plateau pika and SD rat. In conclusion, plateau zokor and plateau pika adapt better to hypoxia than SD rat by increasing the SV of mitochondria, MVD and content of Mb in the cardiac muscle. However, the parameters of mitochondria in the two high-altitude animals are different possibly because of the differences of habitats and habits.
Adaptation, Physiological
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Animals
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Heart
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physiology
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Hypoxia
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metabolism
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Isoenzymes
;
metabolism
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L-Lactate Dehydrogenase
;
metabolism
;
Lagomorpha
;
physiology
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Rats
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Rats, Sprague-Dawley
;
Tibet
6.Effects of lemon peel extracts on lactate dehydrogenase and sucrase activity of Streptococcus mutans.
Xiang-yu ZHANG ; Zhi-fen YU ; Da-zhao WANG ; Ying LIU ; Mao-ding GUO
Chinese Journal of Stomatology 2010;45(12):754-758
OBJECTIVETo investigate the effect of lemon peel extracts (LPE) on the activity of lactate dehydrogenase and sucrase of Streptococcus mutans (Sm).
METHODSAfter serial dilution with trypticase soy broth (TSB) medium containing 2% glucose, LPE was used as the experimental group, and TSB without LPE as the control group. Sm was added to each group, which was then cultured for 6, 18, 24 and 48 hours in the anaerobic tank. The activity of lactate dehydrogenase(LDH) was measured with the method of oxidation of reduction coenzymeIand the pH value of the culture solution was also detected. The activity of the sucrose was determined with the method of coloration of 3,5-dinitrosalicylic acid.
RESULTSThe activity of LDH, sucrase and the changes of solution pH were decreased with the increase of the concentration of LPE (P < 0.01). The activity of LDH were declined from (0.8025 ± 0.0913) × 10(3) U/L to (0.2099 ± 0.0283) × 10(3) U/L; the activity of sucrase were declined from (-0.0107 ± 0.0003) × 10(3) U/L to (-0.0078 ± 0.0002) × 10(3) U/L; the ΔpH were declined from (2.8067 ± 0.0404) to (2.5033 ± 0.0416) (24 h results). The differences were significant between experimental groups and the control group (P < 0.01), and there were also significant differences among experimental groups with different LPE concentration (P < 0.01). The inhibitory effect of acid generation and lactate dehydrogenas' activity of Sm were positively correlated (P < 0.01).
CONCLUSIONSLPE can inhibit the activity of lactate dehydrogenase, sucrase and the acid production capacity of the Sm in a dose dependent manner. The inhibitory effects in logarithmic phase is stronger than that in other phases of growth cycle.
Citrus ; chemistry ; Glucose ; L-Lactate Dehydrogenase ; metabolism ; Lactic Acid ; Plant Extracts ; pharmacology ; Streptococcus mutans ; enzymology ; Sucrase ; metabolism
7.Changes in bronchoalveolar lavage fluid of rats exposed to metal grinding dusts.
Yonghui WU ; Lianming WANG ; Zhongyi ZHANG ; Huiming SUN ; E J LOVE
Chinese Journal of Industrial Hygiene and Occupational Diseases 2002;20(6):446-448
OBJECTIVETo investigate the effects of composite grinding dusts on rat respiratory system.
METHODSRats were administrated with grinding dusts by intratracheal injection. After 2 weeks, the total numbers of cells, the percentage of differential cell, the survival rate of cell, and the activity of lactate dehydrogenase(LDH) and alkaline phosphatase(ALP) in bronchoalveolar lavage fluid were analyzed.
RESULTSAlong with increasing concentration of grinding dusts, the total number of cells in lavage also increased, and was more than that in quartz group. Compared with control group, the percentage of neutrophil in lavage of rats treated with grinding dust and quartz significantly increased and meanwhile that of macrophage significantly decreased[PMN: quartz group (33.83 +/- 4.54)%; grinding dusts group (26.50 +/- 3.99)%, (36.00 +/- 3.58)%, (38.00 +/- 2.10)% at 10, 25, 50 mg/ml respectively. Macrophages: quartz group (62.17 +/- 4.54)%; grinding dusts group (70.83 +/- 3.66)%, (60.83 +/- 2.14)%, (58.17 +/- 2.48)%] while those in control group were (2.83 +/- 0.75)%, (95.67 +/- 1.21)% respectively. The cell survival rate in lavage in control group was 80%, but that in grinding dust and TiO2 group significantly decreased(P < 0.01). The activity of LDH and ALP in all rats treated with dusts obviously increased, and there was significant difference compared with control group(P < 0.01 or P < 0.05). Furthermore, there was significant difference between grinding dust group and quartz group, and between grinding dust group and TiO2 group respectively.
CONCLUSIONMetal grinding dust is very harmful to rat's lung cells and may cause fibrogenesis in the lungs.
Alkaline Phosphatase ; metabolism ; Animals ; Bronchoalveolar Lavage Fluid ; chemistry ; cytology ; Dust ; L-Lactate Dehydrogenase ; metabolism ; Lung ; pathology ; Metals ; Rats
8.Expression of a Lactobacillus casei L-lactate dehydrogenase mutant in Pichia pastoris for asymmetric reduction of phenylpyruvate.
Ting ZHANG ; Jianfang LI ; Die HU ; Chuang LI ; Bochun HU ; Minchen WU
Chinese Journal of Biotechnology 2020;36(5):959-968
To improve the productivity of L-phenyllactic acid (L-PLA), L-LcLDH1(Q88A/I229A), a Lactobacillus casei L-lactate dehydrogenase mutant, was successfully expressed in Pichia pastoris GS115. An NADH regeneration system in vitro was then constructed by coupling the recombinant (re) LcLDH1(Q88A/I229A) with a glucose 1-dehydrogenase for the asymmetric reduction of phenylpyruvate (PPA) to L-PLA. SDS-PAGE analysis showed that the apparent molecular weight of reLcLDH1(Q88A/I229A) was 36.8 kDa. And its specific activity was 270.5 U/mg, 42.9-fold higher than that of LcLDH1 (6.3 U/mg). The asymmetric reduction of PPA (100 mmol/L) was performed at 40 °C and pH 5.0 in an optimal biocatalytic system, containing 10 U/mL reLcLDH1(Q88A/I229A), 1 U/mL SyGDH, 2 mmol/L NAD⁺ and 120 mmol/L D-glucose, producing L-PLA with 99.8% yield and over 99% enantiomeric excess (ee). In addition, the space-time yield (STY) and average turnover frequency (aTOF) were as high as 9.5 g/(L·h) and 257.0 g/(g·h), respectively. The high productivity of reLcLDH1(Q88A/I229A) in the asymmetric reduction of PPA makes it a promising biocatalyst in the preparation of L-PLA.
L-Lactate Dehydrogenase
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genetics
;
Lactobacillus casei
;
enzymology
;
genetics
;
Phenylpyruvic Acids
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metabolism
;
Pichia
;
genetics
;
Recombinant Proteins
;
genetics
;
metabolism
9.Effects of cocaine on activities of ATPase, LDH and SDH in mouse splenocytes.
Wen-ping SUN ; Yan-xu LU ; Xiao-yu ZHANG ; Wei-wei TANG ; Qing-yu HUANG
Journal of Forensic Medicine 2010;26(2):81-83
OBJECTIVE:
To examine the effects of cocaine on the activities of ATPase, LDH and SDH in cultured mouse splenocytes in vitro.
METHODS:
The ATPase, LDH and SDH activities in mouse splenocytes were detected at day 7 after continuous culturing the mouse cells exposed to cocaine hydrochloride in final concentration of 10, 20 and 100 microg/mL in vitro.
RESULTS:
The activities of ATPase, LDH and SDH in mouse splenocytes exposed to cocaine hydrochloride in final concentration of 10, 20 and 100 microg/mL were significantly decreased after continuous culturing for 7 days.
CONCLUSION
The present study demonstrated that cocaine could inhibit the activities of ATPase, LDH and SDH in cultured splenocytes in vitro.
Adenosine Triphosphatases/metabolism*
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Animals
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Cells, Cultured
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Cocaine/pharmacology*
;
Dose-Response Relationship, Drug
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L-Lactate Dehydrogenase/metabolism*
;
Male
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Mice
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Mice, Inbred Strains
;
Spleen/enzymology*
;
Succinate Dehydrogenase/metabolism*
10.Effects of leukocyte elimination before storage on quality of red blood cell concentration.
Hai-Bao WANG ; Jing-Han LIU ; Zi-Lin LIN ; Xi-Jin LI ; Liu-Cai LÜ ; Xi-Lin OUYANG ; Ning HUANG
Journal of Experimental Hematology 2003;11(6):650-653
The objective of this study was to explore the possible effects of leukocyte elimination by filteration before storage on the quality of red blood cell concentrations (RCC) that prepared through two procedures. Eight units of red blood cell concentrations derived from whole blood after plasma separated (RCC1) and eight units of red blood cell concentrations derived from whole blood after platelet-rich plasma separated (RCC2) were divided randomly into filtered group and control group respectively. The RCC of filtered group were filtered by leukocyte deplete filter before storage. The control group didn't have any other treatments. These two groups were stored for five weeks at 4 degrees C according to AABB standard. Mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC) and plasma concentration of K(+) and lactate dehydrogenase (LDH), free hemoglobin (FHb), adenosine triphosphate (ATP) of red blood cell of all RCC were evaluated weekly, and bacteria contamination of all RCC was also detected after five weeks of storage. The results showed that there was no difference of MCV, MCH and MCHC and ATP level of red blood cell in all RCC of two groups, the ATP of red blood cell was lower than the control group on week 4 and 5. The average concentration of K(+) of the filtered group was less than the control group. The differences are significant except that of RCC1 stored till the third week. The plasma LDH concentration of filtered group was less than the control group, and the differences were exacerbate during the storing time prolonged. FHb release in the filtered group of RCC2 was significant less than that of control, but no significant difference was found between the two groups of RCC1. It was concluded that leukocyte elimination by filter before storage could be benefit to RCC preservation.
Adenosine Triphosphate
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metabolism
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Blood Component Removal
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Blood Preservation
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Erythrocytes
;
physiology
;
Filtration
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Humans
;
L-Lactate Dehydrogenase
;
blood
;
Leukocytes
;
Potassium
;
blood